Game Meat Hygiene in Focus Microbiology Epidemiolo

Game meat hygiene in focus

Microbiology, epidemiology, risk analysis and quality assurance
edited by:
P. Paulsen
A. Bauer
M. Vodnansky
R. Winkelmayer
F.J.M. Smulders
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4 Y F P M W L I V W
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Preface
Game meat is consumed world-wide. In most regions, it contributes only a small part to
the overall meat and food supply, but for reasons of animal welfare and sustainability it is
sometimes considered an alternative for meat from farmed animals. Despite differences in
game species, ante mortem conditions (free-range or fenced; wild or semi-domesticated),
hunting or harvesting procedures and further handling of the carcass, there are common
requirements as regards meat safety and quality. Whereas meat safety and shelf life have
been an issue in game meat for export/import for a long time, primary production, domestic
supply and direct supply to the consumer have recently been addressed by legislation and
these sectors still present unresolved questions and challenges.
‘Hygiene’ is commonly defined as all measures that promote and preserve health. It is often
tacitly assumed that hygiene deals with the management of biological, chemical and physical
hazards. However, considering that ‘health’ is ‘a state of complete physical, mental and social
well-being and not merely the absence of disease or infirmity’ (WHO, 1948), or, in a more
recent definition ‘... a resource for everyday life ...’ and ‘ ... a positive concept emphasizing
social and personal resources, as well as physical capacities’ (WHO, 1986), it seems entirely
justified to include under ‘hygiene’ also aspects of sensory meat quality, ethics and sustainable
production, as these contribute to ‘social well-being’.
Having regard of the above-mentioned considerations, a consortium of food hygienists has

established a forum that provides a platform for all questions of game meat hygiene, safety
and quality, consequently termed ‘International Research Forum on Game Meat Hygiene’,
IRFGMH. A major task of this forum is to promote research in all fields of game meat hygiene,
quality and safety, and allow exchange of information in the form of biannual international
conferences and by means of electronic communication.
The first conference of the IRFGMH was held at Brno, Czech Republic, in June 2009. The
hospitality and professionalism of the local organizers, the University of Veterinary and
Pharmaceutical Sciences in Brno, and, in particular, the Central European Institute for
Wildlife Ecology, substantially contributed to the success of this conference.
This book includes chapters authored by key contributors and synopses of other contributions
by participants. Based on their conference presentations, contributors were invited to update
their contributions where necessary and their papers have gone through an extensive review
process. The content is grouped into 4 main sections, viz. ‘hygiene and microbiology’,
‘epidemiology’, ‘risk assessment and management’ and ‘muscle biology and meat quality’.
The contributions represent research outputs, opinions and experiences of experts from 8
European countries, as well as from South Africa, a major game meat exporter.
Game meat hygiene in focus 7

This volume is the first of a book series on safety and quality assurance along the game meat
chain, following an approach ‘from forest to fork’ and is targeted at scientists in academia
and industry, graduate students as well as at governmental officials in veterinary public health
and food safety.
Last but not least, we would like to acknowledge the ‘Verein Grünes Kreuz’, Austria for their
financial support which made this publication possible.
Vienna and Brno, February 2011
The editors
Contents
Preface 7
Section 1 – Hygiene and microbiology
Key contributions
Hygiene and microbiology of meat from wild game: an Austrian view 19

Peter Paulsen
Summary 19
1. Introduction 19
2. Microbiology of meat from wild game 24
4. Conclusions 32
References 33
Essential food safety management points in the supply chain of game meat in
South Africa 39
Johan L. Bekker, Louw C. Hoffman and Piet J. Jooste
Summary 39
1. Introduction 39
2. Methodology 42
3. Game meat supply chain 42
4. Conclusions 60
Acknowledgements 61
References 61
Game harvesting procedures and their effect on meat quality: the Africa
experience 67
Diana L. van Schalkwyk, Louw C. Hoffman and Liesel A. Laubscher
Summary 67
1. Introduction 67
2. Harvesting techniques 71
3. Harvesting operations for meat export 79
4. Conclusions 87
References 88
Zoonotic diseases and direct marketing of game meat: aspects of consumer

safety in Germany 93
Niels Bandick and Andreas Hensel
Summary 93
1. Introduction 93
2. Importance of hunting and wild game consumption in the society 93
3. Safety strategies on game meat hygiene 94
4. Food chain and traceability 95

5. Pathogens and health risks 95
6. Conclusions 99
References 99
Other contributions
Dog bites in hunted large game: a hygienic and economical problem for game
meat production 101
João R. Alberto, João P. Serejo and Madalena Vieira-Pinto
Summary 101
1. Introduction 101
2. Materials and methods 102
3. Results 102
4. Discussion and conclusions 103
References 104
Verocytotoxigenic Escherichia coli (VTEC) in wild ruminants in Germany 107

Andrea C. Bartels and Michael Bülte
Summary 107
1. Introduction 107
2. Occurrence of VTEC in wild ruminants in Germany 108
3. Ports of entry of VTEC in game meat 109
References 110
Microbial quality of venison meat at retail in the UK in relation to

production practices and processes 113
Philip J. Richards, Siyu Wu, David B. Tinker, Mary V. Howell and
Christine E.R. Dodd
Summary 113
1. Introduction 113
2. Material and methods 114
3. Results 115
4. Conclusions 116
References 117
Detection of Alaria spp. mesocercariae in game meat in Germany 119

Katharina Riehn (neé Möhl), Knut Große, Ahmad Hamedy and Ernst Lücker
Summary 119
1. Introduction 119
3. Results and discussion 122
4. Conclusions 124
Acknowledgements 124
References 125
10 Game meat hygiene in focus

Hygiene management systems for commercial game harvesting teams in
Namibia 127
Diana L. van Schalkwyk and Louw C. Hoffman
Summary 127
1. Introduction 127
2. Hygiene management systems 128
3. Conclusions 129
References 129
Salmonella spp. in wild boar (Sus scrofa): a public and animal health concern 131
Madalena Vieira-Pinto, Luísa Morais, Cristina Caleja, Patrícia Themudo,
José Aranha, Carmen Torres, Gilberto Igrejas, Patrícia Poeta and
Conceição Martins
Summary 131
1. Introduction 131
4. Conclusions 134
References 135
Preliminary results indicating game meat is more resistant to

microbiological spoilage 137
Louw C. Hoffman and Leon M.T. Dicks
Summary 137
1. Introduction 137
2. Materials and methods 138
References 139
Section 2 – Epidemiology
Key contributions
Trichinellosis in wild and domestic pigs and public health: a Serbian perspective 143
Sava Buncic and Milorad Mirilovic
Summary 143
1. Introduction 143
2. Main characteristics of disease 144
3. Current trichinellosis status 147
4. Main principles of Trichinella controls 153
5. Conclusions 155
References 156

Influence of climate change on diseases of wild animals 157
Armin Deutz, Thomas Guggenberger and Johann Gasteiner
Summary 157
1. Introduction 157
2. Tularaemia in Austria 161
4. Results 162
5. Loss of habitats in Alpine regions 165
6. Conclusions 169
References 170
Dynamics of infectious diseases according to climate change: the Usutu virus

epidemics in Vienna 173
Franz Rubel and Katharina Brugger
Summary 173
1. Introduction 173
2. The epidemic model 175
3. Parameter estimation 177
4. Climate forcing of the observational period 2001-2005 182
5. Short-term simulation results for the period 2001-2005 182
6. Climate forcing for the extended period 1901-2100 185
7. Long-term simulation results for the period 1901-2100 188
8. Conclusions 190
References 191
Appendix A The USUV model 195
Appendix B R-Source-code for the USUV model 197
Other contributions
The utility of GIS in studying the distribution of Bovine Tuberculosis in wild

boar (Sus scrofa) and red deer (Cervus elaphus) in Central Portugal 199
João R. Alberto, José M. Aranha, João P. Serejo, Alice Amado and
Madalena Vieira-Pinto
Summary 199
1. Introduction 200
4. Conclusions 204
References 204

Section 3 – Risk assessment and management
Key contributions
Risk Management of game: from theory to practice 209

Milorad Radakovic and John Fletcher
Summary 209
1. Introduction 209
2. Hazards in game meat (wild and farmed) 211
3. Risk Management: from theory to practice 213
4. Wild and farmed game meat production 216
5. Practical Risk Management in game meat 217
6. Conclusions 220
References 221
The monitoring of selected zoonotic diseases of wildlife in Lombardy and

Emilia-Romagna, northern Italy 223
Simone Magnino, Matteo Frasnelli, Massimo Fabbi, Alessandro Bianchi,
Maria Grazia Zanoni, Giuseppe Merialdi, Maria Ludovica Pacciarini and
Alessandra Gaffuri
Summary 223
1. Introduction 224
2. Study area 225
3. Diseases 226
4. Conclusions 236
References 237
Assurance of food safety along the game meat production chain: inspection
of meat from wild game and education of official veterinarians and ‘trained
persons’ in Austria 245
Rudolf Winkelmayer, Peter-Vitus Stangl and Peter Paulsen
Summary 245
1. Introduction 245
2. Inspection system for wild game in Austria (excl. Trichinella) 246
3. Trichinella inspection 252
4. Conclusions 256
References 257

Other contributions
Structure and legal framework for the direct local marketing of meat and
meat products from wild game in Austria: the Lower Austrian model 259
Verena Fettinger, Frans J.M. Smulders and Peter Paulsen
Summary 259
1. Introduction 259
2. Current legal framework in Austria 260
3. Achievements to date 260
4. Conclusions 264
References 265
Approaches to game hygiene in the province Belluno (Italy): from training to

meat microbiology 267
Carlo V. Citterio, Patrizia Bragagna, Enrico Novelli and Valerio Giaccone
Summary 267
1. Passive sanitary surveillance 267
2. Game meat microbiology 269
References 270
Section 4 – Muscle biology and meat quality
Key contributions
The muscle biological background of meat quality including that of

game species 273
Peter Hofbauer and Frans J.M. Smulders
Summary 273
1. Introduction 273
2. Muscle structure, composition and function 274
3. Post mortem muscle physiology; the conversion of muscle to meat 276
4. Major sensory characteristics of meat 278
5. Some data on game meat species 288
6. Conclusions 292
References 293
Muscle biological and biochemical ramifications of farmed game husbandry

with focus on deer and reindeer 297
Eva Wiklund and Frans J.M. Smulders
Summary 297
1. Introduction 297
2. Production systems for venison 298
3. Impact of production systems on venison quality 301

4. Ethics and image of venison 310
5. Conclusions 310
References 311
Other contributions
A summary of methods to assess major physical-chemical and sensory

quality traits of fresh (whole tissue) meat 315
Summary 315
1. Introduction 315
2. pH and temperature decline 315
3. Waterholding capacity (drip loss; cooking loss) 316
4. Fresh meat colour 318
5. Tenderness traits 320
6. A remark on problems associated with sample size: animal species
differences 323
References 323
Evaluation of some parameters of post mortem changes of pheasant

(Phasianus colchicus) 325
Jozef Nagy, Peter Paulsen, Peter Popelka, Peter Lazar, Jaroslav Soroka,
Valent Ledecký, Slavomír Marcinčák, Beáta Korénekova and Pavel Bystrický
Summary 325
1. Introduction 326
4. Conclusions 330
References 331
Biographies of key contributors 333
Index 341

Section 1
Hygiene and microbiology
Key contributions
Hygiene and microbiology of meat from wild game:
an Austrian view
Peter Paulsen
Institute of Meat Hygiene, Meat Technology and Food Science, Department for Farm Animals
and Veterinary Public Health, University of Veterinary Medicine Vienna, Veterinärplatz 1,
1210 Vienna, Austria; peter.paulsen@vetmeduni.ac.at
Summary
The food chain for meat from wild game in Central Europe differs from that of farm animals
in the mode of killing (large game: free bullet in head, neck or anterior chest; small game:
multiple shot pellets), conditions of evisceration, storage of carcasses (skin-on) and post and
ante mortem inspection. Poor placement of shots, efflux of ingesta or feces during evisceration,
delayed or insufficient cooling are known to compromise the microbiological condition of
meat. Conditions at primary production level are poorly standardised which may explain
the wide range: Total Aerobic Counts (TAC) on meat cuts from wild game. Counts in the
order of 3-4 log cfu/cm2 can be achieved when Good Hygiene Practice is strictly adhered
to, but values of >8 log cfu/cm2 are also reported. A number of studies report prevalences of
pathogenic bacteria in intestines, tonsils or on skin, but relevant literature on the presence
of a pathogen in the life animal and to what extent it will result in contamination of meat
cuts is scarce. When meat from wild game is placed on the market the same way as that from
farm animals, it is reasonable to demand that similar bacterial limits are set as performance
objectives. This could be, for instance, for large game, that TACs must not exceed 6 log, and
E. coli not 2 log cfu, per cm2 on exposed muscle surfaces of skin-on carcasses arriving at game
handling establishments and per g for meat cuts at retail. It is concluded that optimising the
primary production level is the key to improving safety and shelf life of wild game meat.
Keywords: wild game meat, pathogenic bacteria, spoilage, good hygiene practice
1. Introduction
1.1 Hygiene and the game meat chain: how things developed
Hunting has been the very first way to provide meat for human nutrition, but its relative
contribution to meat supply has – with few local exceptions – decreased. In the Middle Ages
still regarded as ‘substantial’ for overall meat supply (Lerche, 1957; Kerschagl, 1964), meat
from hunted wild game constitutes only ca. 1% of today´s average consumption of meat and
meat products in Austria (Anonymous, 2010).
Practical hunting can be quite different between countries and continents, with respect to
game species and hunting techniques. This contribution will thus focus on the situation in
Austria and Germany, where hunting has a long tradition and a large number of hunters is
P. Paulsen et al. (eds.), Game meat hygiene in focus, 19

DOI 10.3920/978-90-8686-723-3_1, © Wageningen Academic Publishers 2011
Peter Paulsen
more or less actively involved in meat production. The main differences of the game meat
chain as compared to meat production from farm animals are:
• wild game is hunted either in the wild or in large fenced areas;
• for large game, hunters take either fixed positions and wait for the animal (‘still hunting’)
or they stalk animals; another option are drive hunts/battues, where animals are disturbed
and are awaited by hunters;
• for small game, drive hunts are most common;
• animals are killed by free-flying bullets/shot pellets;
• in case of single bullets the shot wound can be located in the anterior chest, head or neck;
• in case of multiple shot pellets, the wounding pattern is highly variable;
• via the shot wound, bacteria can be introduced in the blood stream;
• in some instances, evisceration can be done only with some hours delay, and for small
game, even later;
• evisceration often done ‘on the spot’ with restricted access to clean water;
• traditions that may interfere with modern food hygiene standards;
• cooling of skin-on carcasses;
• ante and parts of post mortem inspection assigned to hunters/’trained persons’;
• for most species, the meat production is seasonal, with peak-periods.
The implications for meat inspection are discussed elsewhere in this volume (Winkelmayer
et al., 2011) as are the ramifications with respect to local trade (Fettinger et al., 2010a, 2011).
The following subchapters will focus on the microbiology of game at the different steps in
the food chain in the light of food safety and quality.
Although game meat has served as food for such a long time, it has not been a main concern
of food hygienists for quite a long time. Admittedly, the need for a meat inspection system
for game has been an issue before 1910, e.g. Kniewallner (1969) cites one reference from
1907, and the rationale for and feasibility of such inspection systems have been discussed in
the following decades (e.g. Olt, 1943). It can be argued that processes as performed in game
handling establishments (skinning, deboning, etc.) are basically the same as in slaughter/
cutting facilities for farm animals, but the primary production area (including killing,
evisceration, transport, cooling), which is different from that of slaughter animals received
little attention.
Reviewing food hygiene literature, the Austrian food codex in its first edition (Anomymous,
1912) merely describes briefly the various species of meat game, differentiates ‘haut-gout’
(defined as a sort of ‘acidic fermentation’ with minor H2S formation only) from spoilage
and mentions that the current practice of shipping unviscerated, still warm carcasses is
not acceptable. A food inspector´s textbook of 1930 essentially repeats these statements
(Messner, 1930). In these times, the main Austrian textbook for veterinary meat hygienists
did not address game meat at all (Postolka, 1903), or, in its last edition (Postolka, 1922) only
reviewed selected aspects (e.g. identification of species, age and gender, and spoilage), but – in
contrast to slaughter of farm animals – provided no specific information on the preferability
of certain handling practices during evisceration, cooling, and transport. In German meat
inspection textbooks by Ostertag (e.g. Ostertag, 1910), game meat was dealt with similarly.
Lerche (1957), from a food hygienist´s view, provided more information on game meat and

An Austrian view
also suggested cooling temperatures for game carcasses of 0 °C. He also questioned the term
‘haut-gout’: originating from an era with a lack of cooling facilities, ‘haut-gout’ was often
used to present the onset of spoilage in an appealing light. Interestingly, a number of authors
mention that meat from wild game would be more resistant to microbial spoilage, which they
in part attributed to low pH and antibacterial activity of residual blood in muscles (Postolka,
1922; Ostertag and Schönberg, 1955; Lerche, 1957).
From the 1960´s onwards, there was a continuous discussion about inspection and hygiene
of wild game meat (e.g. Englert et al., 1965; Prändl, 1976; Raschke, 1976), in part driven by
the need for inspection of venison imported from e.g. New Zealand or hares from Argentina,
and later by the discrepancy between imported venison (which was subject to inspection) and
domestic venison (for which only Trichinella inspection was mandatory) (e.g. Kniewallner,
1969; Greuel, 1979; Riemer and Reuter, 1979). Later, the tendency to prepare meat from wild
game ‘rare’ or ‘medium’ prompted food hygienists for intensified studies (Ring et al., 1988;
Deutz et al., 2000).
In other words, scientific studies and food control focused on the final steps of the game meat
chain for quite a long period, although it was principally agreed that meat hygiene principles
have to be followed by hunters (Kniewallner, 1969).
In the area of primary production, some, although slow, developments could be observed. A
brief view on major training textbooks published over the last 50 years can substantiate this.
Originally, emphasis was put on the description of evisceration methods (carcass lying on the
ground; facultative removal of thoracic organs) and it was stressed that evisceration should be
done as soon as possible for large game (Voß, 1962) or all game species (Anomymous, 1956),
but that carcasses should be opened only to an extent which would minimise contamination
during further transport. It was pointed out that carcasses should be allowed to cool, i.e. to
reach ambient temperature (within 12-24 h) before being transported. Cleaning of the body
cavities should be done with a cloth or even blood (sic!), but not with water. To keep away or
repel insects, storage under good ventilation was recommended, also swabbing of the body
cavities with vinegar. For long-term storage, the skin-on carcass (for small game also un-
eviscerated) could be stored deep-frozen (Voß, 1962).
In the 1950´s and early 1960´s, these recommendations were oriented towards the possibilities
in practice: transportation of game from forest/field to game larders, etc. was generally
considered difficult, and refrigeration facilities were scarce and not easily accessible or
affordable for individuals. Recommendations also included traditional elements. It is
interesting to observe that these were repeated in the major Austrian training books for
hunters (‘Der Jagdprüfungsbehelf’) for more than 25 years and 7 editions with only minor
modifications (Voß, 1962; Anomymous, 1989). The 12th edition of the textbook of 1992
(Anonymous, 1992) was the first to explicitly state that for game put on the market, even in
the absence of formal inspection procedures, the responsibility of the hunter as food producer
was applicable as in any other sector of food production. It also adopted parts of the EU wild
game directive (EC, 1992) and stressed – inter alia – the necessity of proper cooling and
the maintainance of low temperatures with specified maxima, and Trichinella-inspection
of wild boar carcasses. The following editions up to the current one were then based on

Peter Paulsen
the Austrian implementation of the wild game directive and, from 2004 onwards, on EU
legislation (‘hygiene package’). Education of hunters and ‘trained persons’ in order to comply
with the ‘hygiene package’ and the role of hunters as ‘food business operators’ are discussed
in detail elsewhere in this volume (Fettinger et al., 2011 and Winkelmayer et al., 2011).
In summary, improvements in hygiene of the slaughter of farm animals and in cooling

facilities as established from the end of the 19th and first half of the 20th century onwards have
found their way into game handling establishments, but there was some delay in adoption by
primary producers of game meat. The next section will discuss this issue.
1.2 Separating science from tradition: best, good and common practice today and
yesterday
Tradition plays a strong role in hunting, and this also holds true for handling of hunted game.
Critical remarks on hunting traditions having negative consequences for meat hygiene have
been made by Kniewallner (1969) and Riemer and Reuter (1979) and it has been emphasised
that cooperation and compliance of hunters are needed to improve game meat hygiene.
For example, tradition required that during evisceration (‘red work’) sleeves of shirts should
not be rolled up and protective gloves should not be used (e.g. Anonymous, 1956; Voß, 1962),
which was abandoned in the 1970´s (e.g. Anonymous, 1977). The still widely practised ritual
of post-hunt presentation of large and small game (‘Streckenlegung’) is not without hygienic
complications, in particular at ambient temperatures >12 °C and when already eviscerated
carcasses are arranged on the ground (Winkelmayer, 2009).
Current EU legislation suggests that ‘guides to good practice’ should be developed by the food
sectors, and with respect to meat there are a number of textbooks in Germany and Austria
which more or less explicitly deal with ‘good practice’. Among some of the more recent are,
e.g. Kujawski (2007), Martini (2008), and Winkelmayer et al. (2007).
Today’s definition of good or best practice starts before the actual hunting event, and even
before the ante mortem examination immediately before killing (Bandick and Ring, 1996).
It is not only necessary to provide an adequate infrastructure for hygienic evisceration and
refrigeration, but also to better organise drive hunts and battues to facilitate collecting and
evisceration of large numbers hunted game in due time (e.g. Deutz, 2000; Deutz et al., 2006).
Correct placement of shots and immediate death of the animal is relevant not only for animal
welfare (Winkelmayer, 2009), but will also facilitate collecting of the game and allow early
evisceration (Winkelmayer et al., 2005). For large game and under conditions prevailing in
Austria (i.e. numerous semi- or non-professional hunters, the need for short flight distances
due to small hunting areas, the need to preserve head and antlers of male specimens as
trophies), the correct aiming point was defined as an area delineated by the triangle: (1) caudal
contour of shoulder blade; (2) humerus; (3) line from olecranon to the caudoventral edge of
the scapula. This aiming point is slightly more cranial than traditional aiming points. By this,
at the expense of damaging shoulder muscles, the risk of abdominal lesions (Winkelmayer
et al., 2005) and subsequent microbial contamination (Paulsen et al., 2003) was minimised.

An Austrian view
For large game, evisceration must be done without ‘undue delay’, e.g. Winkelmayer et al.
(2008) specify a maximum of 3 h, a time span which is entirely realistic (see Table 1). The
rationale is not only limiting the risk of microbial spread, but also to prevent hot, large
carcasses (wild boar, red deer) from undergoing non-bacterial spoilage (‘stickige Reifung’,
‘stifling maturation’). Recommendations to open the abdominal cavity only to a small extent
and to make incisions in the axillary region, so as to improve cooling thus allowing delayed
evisceration (‘Lüften’ as mentioned in textbooks from the 1970´s) may have been useful in
times where cooling facilities were either not accessible within due time/distance or simply
did not exist. However, nowadays it is an issue of hygiene management to have operational
refrigeration rooms in suitable distance. Consequently, this practice has been completely
abandoned in favour of evisceration and cooling at the earliest possible occasion. At still
hunts, the majority of carcasses was eviscerated with 30 min. (Brodowski and Beutling, 1998;
Deutz et al., 2000), whereas at drive hunts, there will always be some delay between killing
and evisceration (Deutz et al., 2006), and it a logistic issue to keep this time span <1 h.
A variety of evisceration techniques for large game, with carcasses either lying on the
ground or hanging, is suggested in literature and they all have their advantages as long as
the contamination of the body cavity and muscle tissues with soil and feces is avoided or
minimised and meat inspection is not impaired (see for example Bandick and Ring 1996).
Also, for abdominal shots, different approaches are described (i.e. removal of the diaphragm
and peritoneum or washing out the body cavity without opening the thoracic cavity). To date,
there seem to be no systematic studies on which techniques actually result in a minimised
microbial contamination of meat. For small game, the situation is somewhat complicated as
current practice is to eviscerate small game with some days delay. Admittedly, in large drive
hunts (which are then also societal events) with some hundred or thousand hunted pieces
of wild game, it is a logistic challenge to ensure proper cooling, and timely and hygienic
evisceration immediately after the event. In these instances, 2-3 smaller consecutive drive
hunts with smaller hunting bags could allow a more effective game meat hygiene management.
EU legislation specifies maximum meat temperatures of +7 °C for large and +4 °C for small
game (EC, 2004). However, to keep microbial numbers under control, best practice requires
cooling room temperatures of ca. 0 °C, as already suggested by Lerche (1957), the rationale of
which has more recently been confirmed in studies on carcasses as well as vacuum-packaged
meat cuts (e.g. Paulsen et al., 2005b; El-Ghareeb et al., 2009; Fettinger et al., 2010b). It is not
exactly specified how long the period between killing and onset of refrigeration should be;
national guides specify e.g. 12 h (Winkelmayer et al., 2008). In the 1970´s, it was realistic
Table 1. Time from killing to evisceration, large game (% of carcasses).
<1 h 1-3 h >3 h n= Reference
17.5 75.7 5.8 103 Riemer and Reuter, 1979

82.9 15.8 1.3 234 Brodowski and Beutling, 1998
96.5 2.0 1.5 195 Deutz et al., 2000 (still hunt)
0.0 72.9 27.1 37 Deutz et al., 2006 (drive hunt)

Peter Paulsen
to expect that the majority of carcasses would be cooled to 0-2 °C within 24 h after killing
(Riemer and Reuter, 1979). More recent studies demonstrate that the 12 h requirement is easy
to meet (see Brodowski and Beutling, 1998; Deutz et al., 2000).
A ‘pre-cooling’ phase, as generally recommended for carcasses from slaughter animals in

earlier literature (Postolka, 1922; Ostertag-Schönberg, 1955; Lerche et al., 1957), can be useful
for preserving meat quality in slow-glycolysing muscles, and will allow the evaporation of
moisture and subsequent drying of body cavities (Prändl et al., 1988). This is particularly
useful when the carcasses are subsequently hung in refrigeration rooms with poor ventilation
(as is the case for most small-scale units). This can, however, have negative consequences with
respect to microbial surface contamination (Paulsen and Winkelmayer, 2004). Abandoning
the pre-cooling phase requires professional cooling facilities with strong ventilation and the
ability to remove moisture effectively. The necessity of an easy access to cooling facilities has
been emphasised before (Riemer and Reuter, 1979; Brodowski and Beutling, 1989).
1.3 Panta rhei
‘Good practice’ as defined today can be obsolete within a few years, and the subchapter
above has presented some already historical examples. Quite recent examples are studies
conducted on the shelf-life of unviscerated carcasses of pheasants (Paulsen et al., 2008) and
hares (Paulsen et al., 2005a) stored at +4 °C. The authors concluded that a storage for 3 days
would result in acceptable total aerobic counts, with E. coli number remaining <10 cfu/g
muscle tissue. Consequently, a recommendation to eviscerate refrigerated pheasants and
hares not later than at day 3 post mortem was included in a textbook on game meat hygiene
(Winkelmayer et al., 2007). However, practical experience with marketing of hares in the
year 2009 showed that meat cuts from such carcasses, although not objectionable from a
microbiological point of view, were already tainted, that shelf-life was impaired and thus that
this meat is not easily accepted by supermarkets. Also, fermented sausages made of such meat
were sensorily deterioriated. In this case, it proved to be best practice to eviscerate carcasses
within 24 h post mortem.
2. Microbiology of meat from wild game
This section will – in conformity with EU microbiological food hygiene criteria (EC, 2005) –
focus on total aerobic counts, Enterobacteriaceae and E. coli. Other bacteria, as micrococci,
enterococci and clostridia, which can be found in large numbers on game carcasses (Riemer
and Reuter, 1979; Bandick and Ring, 1995; Bandick and Ring, 1996; Schiefer, 1996) or meat
cuts (Kniewallner et al., 1969; Kobe and Ring, 1992) and also pathogenic and zoonotic bacteria
(Dedek and Steineck, 1994; Schiefer, 1996) will not be dealt with in detail. Also, the reader
should not expect a comprehensive overview on all literature reports published in the last
decades; this book chapter is rather somewhat like a round trip through a meat hygienist´s
book shelf, section ‘hygiene and microbiology’, subsection ‘wild game’.
Large game and small game, differing in the mode they are killed and handled post mortem,
will be discussed separately, where appropriate.

An Austrian view
2.1 Microflora of live game/uneviscerated carcass
Muscle tissue of healthy slaughter animals is generally considered as virtually sterile

(Nottingham, 1982), and the same can be expected for game (Ring et al., 1988; Scherling
and Ring, 1989; Schiefer, 1996; Paulsen et al., 2003). Exposed surfaces, as skin, hair/fleece or
feathers and hooves, and also intestinal content will harbour a number of microorganisms,
and for slaughter animals, the significance of the sources of microbial contamination
has been quantified quite early, e.g. Lawrie and Ledward (2006) cite references from the
period 1939-1941 on that subject. Skin/hide of slaughter animals can harbour from 3.5 to
>10 log cfu/cm2 (Ayres, 1955; Bell, 1997; Small et al., 2005; Antic et al., 2010), which has
always been a matter of concern for food hygienists. Some studies demonstrate that for
cattle, microbial transfer skin/hair to meat will be only 1% or less (see Antic et al., 2010),
but considering the high microbial concentration on hides and the potential presence of
pathogens, microbial transfer is regarded as a significant problem.
Visually ‘clean’ cattle resulted in lower microbial numbers on the carcass (e.g. Reid et al.,
2002). With the emergence of enterohemorrhagic E. coli and the finding that cattle is the main
reservoir, the role of cattle skin was re-examined thoroughly (Reid et al., 2002; Nastasijevic et
al., 2008; Antic et al., 2010) and the efficacy of different decontamination techniques assessed
(Small et al., 2005). These findings should be applicable to wild game also.
Interestingly, there are few data on the microbial load of skin and hair or feathers of wild
game. For example, El-Ghareeb et al. (2009) examined various game bird species (partridge,
pheasant, pigeon, quail) and reported total aerobic counts in the range of 2-6.5 log cfu/cm2
skin/feathers, and median numbers of Enterobacteriaceae, E. coli, and Staph. aureus in the
range of 2-3 log cfu/cm2 . It seems that, to-date, data on enterohemorrhagic E. coli on hides
of wild ungulates are lacking, despite the evidence that such species constitute a natural
reservoir (Bartels and Bülte, 2011). Admittedly, more data have been published on the presence
of pathogens in the digestive tract, e.g. in tonsils and feces (e.g. for wild boar: Wacheck et
al., 2010).
2.2 Contamination at primary production level (shooting, evisceration and

cooling)
2.2.1 Killing
It is well established for farmed game at slaughter, that captive-bolt stunning and sticking
introduce bacteria in the bloodstream (see Lawrie and Ledward, 2006). However, there seems
to be some antibacterial activity in post mortem tissues, as demonstrated by Gill and Penney
(1979): bacteria experimentally inoculated in the bloodstream of animals before slaughter
were disseminated in the body, but their numbers decreased in the following hour, or it took
several hours before microorganisms started to multiply. Recently, and in the light of BSE,
hematogenic dissemination of bacteria inoculated via the captive-bolt wound (Buncic et al.,
2002; Daly et al., 2002; Prendergast et al., 2004) has been studied as a model for CNS spread.
It has become clear that the failure to recover artificially inoculated bacteria may to some
extent be a problem of the concentration of the inoculum. With respect to large game killed

Peter Paulsen
by a shot in the anterior chest, it may be argued that the massive loss of blood through severed
major arteriae will wash away contaminants rather than transport them into deep vessels.
The mode of killing of large game and location of the shot wound have a distinct influence
on the microbial numbers in deep muscle tissues (Lenze, 1977; Ring et al., 1988; Deutz et al.,
2000). Adominal shot lesions are significantly associated with visible contamination of the
abdominal cavity and exposed muscles (Deutz et al., 2000; Paulsen et al., 2003). For carcasses
sampled 12-24 h after killing, Deutz et al. (2006) reported median and maximum TACs of
4.6 and 5.6 log cfu/cm2 for thoracic shot wounds compared to 5.0 and 6.5 log cfu/cm2 for
abdominal lesions, respectively. Also, Paulsen and Winkelmayer (2004) and Langrange and
Schmidt (2005) found higher surface counts for TAC, Enterobacteriaceae and E. coli in game
with abdominal shot wounds, approximately in the same order of magnitude.
Drive hunts for large game are associated with a higher frequency of abdominal shot lesions
and longer time to evisceration. However, it depends on the logistics of evisceration and
cooling whether numbers of indicator bacteria are higher on such carcasses compared to
those obtained from still hunts (Deutz et al., 2006).
2.2.2 Evisceration
Evisceration of carcasses without undue delay is stipulated in legislation, but there is some
evidence that, if intestines are not ruptured or otherwise damaged, bacteria will colonise
muscle tissues quite slowly and that sensory deterioriation (‘greening’) will occur before
microbial spoilage. Gill et al. (1976, 1978) have presented evidence for this hypothesis. In
the case of small game, it was a traditional procedure to hang uneviscerated carcasses for
days to weeks at near ambient temperature, and earlier studies (Barnes et al., 1973; Mead
et al. 1973, 1974) demonstrated that storage of uneviscerated pheasants at temperatures of
10 °C or less for several days does not necessarily result in sensory deterioriation or bacterial
spoilage, unless the intestines are perforated and fecal material is released into the body
cavity (similarly as observed for lambs, Gill et al., 1978). Also, Paulsen et al. (2008) reported
that during the storage of uneviscerated, hunted pheasants at 0 °C and 3-4 °C, total aerobic
counts increased significantly during storage, but the absolute numbers remained below 6
log cfu/g. Whereas E. coli were <1 log cfu/g in muscles of hunted pheasants on day 3 at 4 °C,
counts of up to 3.7 log cfu/g on day 7 at 4 °C indicate a loss of hygienic quality. Therefore, the
authors recommended that hunted uneviscerated pheasants could be stored 3 days at 4 °C,
but not 7 days or more after the hunt.
For hares, Meyer-Ravenstein et al. (1976) reported that eviscerated carcasses stored at 4 °C
are more susceptible to spoilage than uneviscerated ones. From results reported by Heinz et
al. (1977), who compared eviscerated and uneviscerated hares stored at 4 °C and at ambient
temperatures (10-20 °C), it can be concluded that cold storage of uneviscerated carcasses
tends to yield lowest bacterial counts on muscle surfaces, but sample numbers per group are
low (n=5) and results cover a wide range (>2 log for most groups).

An Austrian view
2.2.3 Surface microflora on wild game carcasses in the first 24 h p.m.
Atanassova et al. (2008) sampled 289 freshly shot large game specimens, comprising 127 wild
boar, 95 roe deer and 67 red deer, and reported average TACs of 3.2, 2.9 and 2.6 log cfu/cm2,
respectively. Average Enterobacteriaceae concentrations were 2.2 log cfu/cm2.
Under well-defined conditions (average time to evisceration: 42 min, average time to cooling:
3.1 h), median TAC values for 62 carcasses were 5.4 and 5.6 log cfu/cm 2 for abdominal cavity
and thighs (Deutz et al., 2000). The latter report also mentions higher values on 61 carcasses
sampled at a game handling establishment (5.8 and 6.3, respectively). For Enterobacteriaceae,
E. coli, and Staphylococci, a similar tendency was found.
Riemer and Reuter (1979) sampled 89 large game carcasses and reported TAC <2 log cfu/g
deep muscle tissue in 53% of samples, and for the majority of the remaining samples counts
<4 log cfu/g. Enterobacteriaceae (median <3 log cfu/g) and E. coli (median <2 log cfu/g) were
detected in 11 and 12 of 89 carcasses, respectively.
2.2.4 Microbial contamination and visual assessment of ‘cleanliness’ of carcasses
The relation of visual assessment and microbial contamination of carcass surfaces has been
studied by Deutz et al. (2003) at the level of ‘trained persons’ and by Paulsen et al. (2003) for
veterinary meat inspectors. Deutz et al. (2003) classified carcasses according to contamination
of body cavities and other alterations (such as emaciation, fractures, discolouration) in four
categories, with the two best categories (i.e. not requiring veterinary intervention) being
characterised by TACs <6 log cfu/cm2 and E. coli counts <1 log cfu/cm2. Interestingly, this
correlated rather well with the finding of Paulsen et al. (2003), who examined roe deer
carcasses (n=100) at the entry point of a cutting plant. The results were compared to approval/
condemnation based on visual and olfactory examination performed by the meat inspecting
veterinarian: 88% of the heavily contaminated carcasses (6 or more log cfu TAC/cm 2
abdominal muscle) were condemned or conditionally approved. Despite of microbial surface
contamination, microbial growth was detected in only one core of 100 extensor carpi radialis
muscles. Average TACs of abdominal muscles of 7.6 log cfu/cm2 for visually contaminated
abdominal cavities were found as compared to 5.3 log cfu/cm2 for visually clean abdominal
cavities. Respective numbers for Enterobacteriaceae were 5.1 and 3.5 log cfu/cm2. From swabs
of the thoracic and abdominal cavity, E. coli were isolated from 76 carcasses.
In addition to the previously described factors influencing the exent of microbial contamination,
the time/temperature profile from killing to cooling is often underestimated. Paulsen and
Winkelmayer (2004) found that ‘pre-cooling’ phases (ca. 12 h) at ambient temperatures of ca.
10 °C and 18 °C were associated with median TAC values of 4.1 as compared to 5.7 log cfu/cm2.
Respective numbers for Enterobacteriaceae were 2.5 and 3.5 log cfu/cm2. Subsequent cooling
at 0.4 °C allowed no microbial growth for 96 h, but differences in microbial numbers remained.
This also demonstrates the need for a continuous cool chain.

Peter Paulsen
2.2.5 Microbial limits for carcasses of wild game?
The data presented above raise the question, whether or not microbiological limits for game
carcasses should be defined and if they should be in the range as given for slaughter animals
or higher. To date, there are no standardised sampling methods for wild game (swabbing-
destructive; surface-deep tissue, sampling site, step of the food chain). From various literature
sources, recommendations have been collated in Table 2. Notably, these values are in the upper
range or maximally 1 log unit higher than those for slaughter carcasses.
2.3 Microflora of meat cuts
2.3.1 Microflora of meat cuts: surveys and experimental findings
A number of studies reported that market samples from meat cuts from wild game have higher
microbial numbers than those from farmed animals (Kniewallner, 1969; Baur and Reiff, 1976;
Kobe and Ring, 1992; Bandick and Ring, 1996). As concluded by Schiefer (1996), surface
counts of 7 log cfu/cm2 are not uncommon, but meat cuts from farm animals can harbour
similar numbers of bacteria on their surface (Kobe and Ring, 1992). In deep muscle tissue
from wild game, total aerobic counts have been reported to be quite variable, i.e. from <2 to >5
log cfu/g (Kniewallner, 1969). A later study reports numbers to be significantly higher than in
those from farm animals (Kobe and Ring, 1992; see also Table 3). In particular, the numbers
of Enterobacteriaceae and E. coli (Kobe and Ring, 1992) or coliforms (Kniewallner, 1969)
can be quite high. It is sometimes not easy to compare data, particularly those from older
studies, which used semi-quantitative methods for assessment of bacterial contamination
(e.g. Baur and Reiff, 1976).
Recent studies, conducted at different steps of producing and placing of the market of meat
from wild game, still demonstrate a wide variation of results, but allow the conclusion
that average TACs, Enterobacteriaceae and E. coli numbers of 3-4, 2-3 and 1-2 log cfu/g,
respectively, can be achieved under GHP conditions.
In frozen meat cuts sampled in German game handling establishments, average TACs from
4.0-6.4, and Enterobacteriaceae and E. coli counts of 2.0-6.0 and 1.0-3.1 log cfu/g, respectively,
were reported (Türck, 2008; Wacheck, 2008; Table 4).
Table 2. Suggested microbiological limits for wild game carcasses (log cfu/g), derived from various
literature sources.
TAC Enterobacteriaceae E. coli Based on
6.0 - 2.0 Deutz et al., 2003 (non destructive sampling)

6.0 - - Paulsen et al., 2003 (destructive sampling)
5.0 3.0 2.0 Lagrange and Schmidt, 2005 (destructive sampling)

An Austrian view
Table 3. Microbial counts (total aerobic counts and Enterobacteriaceae) in game meat cuts; studies
conducted in Germany and Austria (data are mean ± standard deviation or median plus range).
Sampling n TAC (log cfu/g) EB (log cfu/g or cm 2) Sensory assessment Reference

or coliforms*
Surface 62 7.33 (4-8.68) 5.48 (2-6.6)* Not objectionable Kniewallner, 1969

Surface 19 6.68±1.01 4.32±1.13 Not objectionable Kobe and Ring, 1992
Surface 35 7.39±1.03 4.67±0.88 Objectionable Kobe and Ring, 1992
Mix 103 6.29 (4.48-7.56) 5.1 (2.3-6.36) Not objectionable Paulsen et al., 2005
Deep tissue 24 4.28 (<2-5.4) 4 (<2-5.38)* Not objectionable Kniewallner, 1969
Deep tissue 19 4.25±1.44 0.71±1.18 Not objectionable Kobe and Ring, 1992
Deep tissue 35 5.72±0.98 1.93±1.49 Objectionable Kobe and Ring, 1992
Table 4. Microbial counts in game meat cuts; recent studies conducted in Germany and Austria (data
are mean* or median plus range).
Samples Condition n Obtained at1 TAC EB E. coli Reference

(log cfu/g) (log cfu/g) (log cfu/g)
Hare frozen 79 GHE 4.0 (2.2-6.2) 2.0 (1.0-3.8) - Türck, 2008

Hare frozen, 85 GHE 4.0 (3.4-8.3) 2.7 (1.0-6.0) - Türck, 2008
larded
Roe deer frozen 44 GHE 5.7 (3.5-7.3) 3.6 (1.0-6.3) 2.3 (1.0-3.8) Türck, 2008
Red deer frozen 49 GHE 4.3 (1.1-6.6) 2.2 (1.0-4.2) 1.7 (1.0-3.7) Türck, 2008
Wild boar frozen 224 GHE 6.0 (2.6-8.6) 3.5 (1.0-5.8) 1.0 (1.0-4.9) Türck, 2008
Various frozen 63 GHE 6.4* (4.3-7.3) 6.0* (4.3-7.0) 3.1* (2.0-4.4) Wacheck, 2008
Various chilled, vac. 30 LSH-sc 4.4 (2.0-7.7) 2.3 (1.0-5.7) 1.0 (1.0-3.5) Fettinger, 2011
Various chilled 43 LSH-training 5.1 (3.0-7.7) 2.7 (2.0-5.7) 1.0 (1.0-3.0) Fettinger, 2011
Roe deer fresh after 49 Experimental 5.7 3.9 2.0 Irschik,
cutting from visually unpublished
contaminated data
carcasses
1 GHE: sampled at game handling establishments; LSH-sc: microbiological self control samples from hunters
supplying directly to consumers; LSH-training: samples taken at training courses for supplying directly to consumers.
For some species, Türck (2008) observed significant differences in microbial contamination
of different meat cuts. Similar observations have been reported by Kniewallner (1969) for
venison and Heinz et al. (1977) for hares.
In 30 vacuum-packed fresh meat cuts obtained from hunters supplying directly to the
consumer, Fettinger (2011) reported median counts of 4.4, 2.3 and 1.0 log cfu/g for TAC,

Peter Paulsen
Enterobacteriaceae and E. coli, respectively. Similar results have been reported from training
courses where hunters were cutting carcasses and packaging meat (Fettinger, 2011).
Meat cuts (n=103) from roe deer stored at 3.5 °C were characterised by 5.8 log cfu/g TAC and
4.0 log cfu/g Enterobacteriaceae (median values, all 132 h post mortem) (Paulsen et al., 2005).
This study included freshly prepared meat cuts as well as vacuum-packed stored meat cuts.
Studies as those mentioned above are based on carcasses where the initial microbial
contamination of the carcass and contributing factors (e.g. time-temperature history, time
to evisceration, visible contamination and location of the shot wound, see above) are not well-
defined. Few studies have been published with the aim of explaining microbial numbers to
be expected under ‘good’ or even ‘best’ practice options.
For roe deer (n=6), Paulsen (2005) reported average TACs of 4.3 and 5.1 log cfu/g for meat
cuts and comminuted meats, when visibly clean carcasses were kept at 3 °C for 3-7 days and
processed under hygienic conditions, whereas hygiene deficiencies resulted in log 2.5-3.5
higher microbial numbers.
For various game bird species, El-Ghareeb et al. (2009) studied the implementation of Good
Hygiene Practice in the cutting process and concluded that median counts of 3-4, <2-2.3
and <1-1.5 log cfu/g are to be expected for TACs, Enterobacteriaceae and E.coli, respectively
for a number of species. Somewhat higher numbers (and subsequent reduced shelf-life) were
reported for pigeon (see Table 5). Fettinger et al. (2010b) studied GHP procedures for meat cuts
of hare and reported similar results. Interestingly, these data are in the same range as reported
previously for frozen hare meat imported from Argentina (Heinz et al., 1977). Pecularities
for small game, as frozen storage of uneviscerated carcasses (Heinz et al., 1977; El-Ghareeb
et al., 2009) and freezing-thawing-freezing cycles during processing of hare (Türck, 2008) are
not necessarily associated with higher microbial contamination of meat, but seem somewhat
archaic in the light of modern food hygiene.
Table 5. Microbial counts in meat cuts from small game, obtained under GHP conditions (data are
median plus range).
Samples n TAC (log cfu/g) EB (log cfu/g) E.coli (log cfu/g) Reference
Hare thighs 20 3.4 (3.0-5.0) 2.6 (2.0-2.9) - Fettinger et al., 2010b

Hare longissimus 6 3.0 (3.0-4.4) 2.9 (2.0-4.3) - Fettinger, unpublished data
Partridge breast 8 2.9 (2.0-3.9) 2.0 (2.0-4.0) <1 (<1-4.1) El-Ghareeb et al., 2009
Partridge thighs 8 3.1 (2.0-5.3) 2.0 (2.0-4.5) <1 (<1-4.5) El-Ghareeb et al., 2009
Pigeon breast 9 4.1 (3.0-6.0) 2.3 (2.0-5.3) 1.7 (<1.0-3.5) El-Ghareeb et al., 2009
Pigeon thigh 9 4.0 (3.3-5.7) 3.3 (<2-5.0) 3.0 (<1-4.0) El-Ghareeb et al., 2009
Quail muscles 25 4.0 (2.0-6.5) <2 (<2-4.2) <1 (<1-4.1) El-Ghareeb et al., 2009
Pheasant breast 32 3.7 (2.0-5.5) 2.0 (2.0-5.3) 1.0 (1.0-4.0) El-Ghareeb et al., 2009
Pheasant thigh 32 3.8 (2.3-5.5) 2.3 (2.0-5.3) 1.5 (1.0-4.0) El-Ghareeb et al., 2009

An Austrian view
Generally, storage temperatures of ca. 0 °C afford keeping microbial numbers on vacuum-
packed meat cuts nearly constant for at least one week (e.g. El-Ghareeb et al., 2009 for wild
birds; Fettinger et al., 2010b, for hares; Irschik, unpublished data, for roe deer). This is of course
not unexpected, but at higher temperatures multiplication of bacteria may be significant, e.g.
as reported by Fettinger et al. (2010b) for hare meat stored at +4 °C (i.e. the maximum legally
allowed temperature).
2.3.2 Sensory assessment and extent of microbial contamination
The relation of microbial numbers to sensory characteristics of wild game meat has been
under study for quite a long time. Kniewallner (1969) and Türck (2008) could not establish a
clear relationship, whereas Kobe and Ring (1992) found that average TACs of >7 log cfu / cm2
surface or >5 log cfu/g deep tissue were associated with sensory deterioriation.
The relation to other parameters indicating protein degradation and thus ageing or the onset
of spoilage was not convincing (Paulsen et al., 2008; Türck, 2008).
Similarly, only a weak (Kniewallner, 1969) or no correlation (Baur and Reiff, 1976; Heinz et
al., 1977; Kobe and Ring, 1992) of pH values to microbial numbers has been found.
2.3.3 Microbial limits for carcasses of wild game?
To date, there are no explicit microbial limits for fresh meat from game, with the exception
of generally applicable food safety criteria as laid down in EU legislation on microbiological
criteria on foodstuffs (EC, 2005). Considering that meat from game is placed on the market
as any other meat from farmed animals, it seems reasonable to adopt limits for fresh meat
from farm animals. These may be included in national legislation or expertises (compiled by
Eisgruber and Bülte, 2006), or part of quality meat programmes, e.g. the ‘Agrarmarkt Austria’
program (AMA, 2010). This approach was followed by two recent studies conducted at game
handling establishments (Wacheck, 2008) and ‘local supply’ level (Fettinger et al., 2010a).
Basically, the evaluation scheme required that food safety criteria (pathogenic bacteria) had
to be fulfilled, whereas for other microbial indicators pertaining to quality and shelf life, the
limits would serve as guidance values only. Exceeding these guidance values would indicate
that improvements in the game meat chain are necessary (example see Table 6 and Fettinger
et al., 2010a). Some recommendations are compiled in Table 7.
The pronounced seasonality in game meat production, and the small-scale of the operation
plants and resulting poor standardisation of processes may contribute to the large variation
of microbial numbers reported for game meat cuts. This seasonality and discontinuous
production has become untypical for most meat species in Austria, maybe with exception of
geese and sheep. However, this mode of production always bears some risk of high microbial
numbers, e.g. Loncaric et al. (2009) reported 7.1±0.9, 3.3±2.2 and 1.3±1.2 log cfu/g for mutton
cuts sampled at retail level in Austria.

Peter Paulsen
Table 6. Microbial limits for game meat and products as used in a pilot trial for hunters supplying to the
consumer (Fettinger, 2010a). Example: vacuum-packed meat (AMA, 2010; Eisgruber and Bülte, 2006).
Category 2 (fresh, vac-packed meat) Tolerance level Absolute limit
Listeria monocytogenes (cfu/g) - <100

Listeria monocytogenes in 25g - n.d.
Salmonella sp. in 25g - n.d.
Sulphite reducing anaerobes (kbE/g) - -
Staphylococcus aureus (cfu/g) - -
Bacillus cereus (cfu/g) - -
TAC (cfu/g) 5,000,000 -

Escherichia coli (cfu/g) 50 500
Enterobacteriaceae (cfu/g) 1000 -
Lactobacillus (cfu/g) 5,000,000 -
Table 7. Suggested microbiological limits for meat cuts from wild game (log cfu/g), derived from various
literature sources.
TAC Lactic acid Pseudomonas E. coli Enterobacteriaceae C. perfringens S. aureus Reference

bacteria
6.7 - - - - - - Bandick and

Ring, 1996
6 6 6 2.7 4 3 Wacheck, 2008
6.7 6.7 - 1.7 3 - - Fettinger, 2011
4. Conclusions
What has been achieved?
In the last 50 years, principles of slaughter hygiene have been adopted for wild game meat
production, with respect to evisceration, cooling and the implementation of ante and post
mortem meat inspection. Under these principles, it has been proven that it is entirely possible
to produce skin-on large game carcasses with TACs not exceeding 6 log, and E. coli <2 log cfu
per cm2 on exposed muscle surfaces. Similarly, it can be expected that the resultant meat cuts
should not contain >6 log TAC and >2 log E. coli/g meat tissue. For small game, traditional
post mortem handling techniques (i.e. hanging of uneviscerated carcasses for several days;
insufficient cooling) are increasingly replaced by rapid cooling and evisceration, and skinning
within 24 h after killing.

An Austrian view
What has been neglected?
Conditions at primary production level are still poorly standardised which accounts for a
wide range of bacterial contamination on carcasses. This may be due to a lack of manual skills,
organisation, infrastructure or simply ignorance. Continuing education of primary producers,
currently on a voluntary basis in Austria, should be emphasised. Likewise, sampling strategies
and techniques along the game meat chain are very diverse, which complicates comparisons
of results.
What needs to be done?
Guides to good practice have been developed according to EU food hygiene legislation, but
the application in practice should be enforced. Despite the variety of hygiene improvements
suggested, stipulated and – in part – adopted in the last decades, average microbial counts
on carcasses and meat have not declined in the order of magnitude expected. Although data
exist on the prevalence of pathogens in the digestive tract, not much is known about the
extent of transfer of these pathogens to the carcass and finally, to meat cuts. This is a crucial
issue, as animals may be symptomless carriers and go undetected during ante as well as post
mortem inspection.
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Peter Paulsen
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Essential food safety management points in the
supply chain of game meat in South Africa
Johan L. Bekker1, Louw C. Hoffman2 and Piet J. Jooste1
1Tshwane University of Technology, Private Bag X680, Pretoria, 0001, South Africa;
bekkerjl@tut.ac.za
2University of Stellenbosch, Private Bag X1, Matieland, 7602, South Africa
Summary
Although the game industry is expanding in all areas, the most essential food safety
management points in the supply chain of South African game meat has not yet been described.
In order to determine the management points it was necessary to determine the supply chain
and then to identify the most essential food safety management points. Information to better
understand the supply chain and relevant food safety management aspects was obtained
through a desk top study, observation of processes from farm to consumer in the local market
as well as during export activities and analysis of questionnaire responses from game farmers,
hunters, butcheries and municipalities. The description of essential food safety management
points in the supply chain can assist policy-makers; law enforcers and the industry to establish
and implement management programmes that will ensure a safe game meat product to the
consumer.
Keywords: supply chain, traceability, sustainable utilisation, hunting, harvesting
1. Introduction
In South Africa, domestic stock has always been a major social, economic and livelihood
support factor to the farmers and the people of the country. However, the process of
desertification in Africa has produced an environment that can no longer support the
increased numbers of domestic cattle economically (Van Schalkwyk, 2004). As a result, South
African farmers were faced in the 1980s with finding alternative economic solutions for their
increasingly marginalised land (Bothma, 2002a). However, from an economic perspective,
extensive cattle farming is more efficient in terms of meat production than game farming
and the total biomass of bovine culled per year is higher than that of game (Dekker and Van
Wyk, 2002). Skinner (1970) also indicated that game animals are unlikely to compete directly
with domestic animals as meat producers as they are not as efficient in converting feed into
live weight and at that time doubts remained concerning disease and their management. In
addition, wild animals were usually seen negatively, e.g. as crop raiders/pests or undesirable
competitors for grazing that could be best used to produce domestic livestock (Bothma, 2002b;
Bothma, unpublished data). On the other hand, Cloete et al. (2007) pointed out that game
ranching can be more profitable, i.e. generate a higher gross margin per hectare than cattle.

Africa has a high density and diversity of large wild herbivores, of which South Africa has
some 300 different species of mammals (ABSA, 2003). In South Africa, as is in other countries
with abundant wildlife, the national economic value of wildlife has traditionally been seen
in terms of its potential to generate revenue through tourism, i.e. photographic tourism by
overseas and local visitor’s and hunting (Campbell et al., 2001). It has since been discovered
that wildlife ranching is more profitable than domestic livestock farming in many parts of
the country, mainly because it produces not only meat and other consumptive and non-
consumptive products (ABSA, 2003) such as accommodation and service fees with a higher
return potential (Dekker and Van Wyk, 2002), but it is starting to become a vehicle for
conservation based rural community development (Bothma, unpublished data). Whilst
conservation and development may be seen as conflicting agenda, attempts are nevertheless
made to bring wildlife conservation closer under the general umbrella of sustainable
development (Meadows et al., 1992; Traffic, 2008). Game farmers in South Africa now play a
key role in the conservation of many game species (Ebedes, 2002). As the economic potential
became clear, many South African farmers moved away from conventional agriculture
towards wildlife production units and as a result of this, wildlife ranching has developed
rapidly to become a multifaceted multi-million Rand industry with tremendous conservation
benefits (Bothma, 2005). South Africa has now become a world leader in the sustainable
extensive conservation and utilisation of game species (Ebedes, 2002). Many cattle farmers
are changing over to game farming (Cloete et al., 2007; Carruthers, 2008) and it is now widely
recognised that game farming is the fastest growing agricultural industry in South Africa.
As a result game numbers seem to have increased and many livestock ranches carry enough
wildlife to allow their commercial exploitation (Carruthers, 2008). The game industry as it
is today provides for a variety of opportunities of utilisation of game that the farmers in the
game industry can benefit from. Figure 1 illustrates the main sectors in the game industry.
Although these different sectors exist, this chapter will mostly discuss the food safety
management points for meat production through harvesting and hunting of wild game. The
term ‘venison’ refers to the meat of cervids, of which the red deer (Cervus elapphus) is the
only true indigenous one in Africa (specifically North Africa). However, since the term ‘game
meat’ is used most frequently in South Africa, it will be used for the purpose of this chapter.
Game industry
Breeding and selling of game
Meat production
Ecotourism
Hunting
Figure 1. Sectors in the game industry.

Essential food safety management points in the supply chain of game meat in South Africa
Game meat has been recognised as a source for meat because:

• Farmers realised that wild ungulates in Africa, when managed properly could out-produce
domestic animals in producing red meat per hectare (Skinner, 1984) and that game
farming could supplement other forms of meat (Skinner, 1970).
• Game meat does not have a high fat content, because there is little marbling in the muscle
(Aidoo and Haworth, 1995; Hoffman, 2008). It is also low in cholesterol, which forms
an essential constituent of all cells of animal origin (Higgs, 2000). Game meat complies
with the nutritional guidelines that place an increasing emphasis on reducing the ratio of
n-6/n-3 polyunsaturated fatty acids in the diet (Hoffman et al., 2005). This was shown to
be favourable with respect to decreasing the risk of coronary heart disease.
• African game meat is considered to be an organic product since the animals are wild
and free-roaming (Hoffman and Wiklund, 2006). The meat is free of growth hormones
and other artificial or chemical substances since no dipping takes place and no fertilisers
or growth stimulants are used in the production system (Radder, 2002; Hoffman and
Wiklund, 2006).
Consequently, alternative sources of meat, such as wildlife are increasing in popularity

(Hoffman, 2003; Webb, 2003). The meat derived from game is consumed by the immediate
family, sold locally in order to provide cash income, or exported as a sought-after delicacy,
especially in Western Europe (Skinner, 1984). A study by Hoffman (2003) indicated that the
majority (73%) of consumers in the Western Cape Province of South Africa had eaten game
meat before. Most of these consumers had purchased the game meat in butcheries and then
prepared it at home. Another study by Hoffman et al. (2003) indicated that game meat is very
popular with tourists (Belgium’s, Germans and Americans), not only as an African product
but as a healthy product. The majority (82%) of tourists were aware of the health benefits of
wildlife meat. Further, 92% of them indicated that they eat game meat when on tour and 88%
when they are at home. Tourists also regard game meat as part of an African experience (Du
Buisson, 2006). In the 2005 season it was estimated that South Africa exported the deboned
meat from 160,000 game carcasses (Hoffman and Wiklund, 2006). However, due to the global
economic climate, this number had decreased due to a lower international market demand
to slightly over 76,000 in 2009 which was lower than the 85,500 of 2008.
The term ‘food supply chain’ is usually reserved for the total supply process from agricultural
production, harvest/slaughter, through primary production and/or manufacturing, to
storage and distribution to retailers or for use in catering and consumer practice (Stringer
and Hall, 2007).
Game meat from game parks, big hunting farms and approved game abattoirs is finding its
way to the consumer’s table through the formal processes or structures. However, informally
hunted game meat has also found a place in the local supply chain since 60-65% of the total
income from game farming is generated from trophy and meat hunting (Van der Merwe,
2005). In identifying essential food safety management points and developing food safety
interventions for game meat control purposes it is critical to understand the dynamics of the
supply chain from the farm to the fork that also include the export of game meat to relevant
countries. The purpose of this chapter is to use the South African supply chain for game meat
as a model to describe essential food safety management points in the supply chain. A clear

understanding of the food safety management points can assist policy makers, law enforcers
and the game meat industry to establish and implement management programmes that will
ensure a safe game meat product to the consumer.
2. Methodology
Information to better understand the supply chain of game meat in South Africa was
obtained through a study that consisted of (1) a desktop review of relevant subject material;
(2) observations made during harvesting and hunting events and (3) the design of separate
questionnaires for the respective target groups. The questionnaires, which were distributed to
local game farmers, hunters, butcheries and municipalities (local authorities) were designed
to obtain information regarding their involvement and related activities in preventative and
legislative measures to ensure safe game meat. In designing the questionnaires international
and local legislation and standards was taken into consideration.
The questionnaires were distributed to game farmers and hunters through their respective
associations which form the official mouthpieces of game farmers and hunters in South Africa.
Questionnaires to butcheries were distributed by senior students registered for the National
Diploma: Environmental Health at the seven universities in South Africa that offer the course.
After clear instructions were provided to students, the questionnaires were distributed to
butcheries during their compulsory in-service training with municipalities. The questionnaire
to municipalities was distributed electronically to the heads of the Environmental Health
departments of all the metropolitan municipalities (n=6) and district municipalities (n=41)
in South Africa.
A total of 139 game farmers, 290 hunters and 361 butcheries completed the questionnaires.
Although two of the six metropolitan authorities submitted their responses per region
in their metropolitan jurisdiction (metro 1 = 8 regions and metro 2 = 7 regions), all six
metropolitan authorities responded. This resulted in a total of 19 questionnaire responses
being received from the metropolitan areas. Twenty eight (68%) district municipalities
completed the questionnaires. Consequently, the total n-value for the municipalities was
47 and a distinction between the metropolitan and district municipalities will not be made.
The quantitative questionnaire responses were coded and analysed using the SPSS statistical
software. Qualitative data were analysed using content analysis. This involved identifying
specific a priori and emergent issues on South Africa that focus on the supply chain of game
meat. Descriptive statistics were calculated to summarise the results in tables by means of
frequencies and summary proportions.
3. Game meat supply chain
The game meat supply chain differs from the supply chain of meat from domesticated animals
in that game animals are killed and partially dressed on the game farm by removing the
viscera of the thorax and abdomen, the head, feet, reproductive organs and lactating udders.

Based on this difference and observations made during the study, Figure 2 was compiled to
depict the supply chain for game meat in South Africa.
Figure 2 clearly illustrates that the supply chain for game meat in South Africa involves several
role-players (including relevant authorities). The essential food safety management points for
the different steps of the supply chain in Figure 2 are systematically described in the following
sections with numbers corresponding to those in Figure 2.
3.1. Primary producers

(game farms) 3.1.1 Suppliers of feed
3.2 Harvesting and hunting

3.2.1. Hunter residences (Including field slaughter depot)
3.5.1 Export
3.4.2 Large scale
3.6 Suppliers of processing materials
3.3.1 Farm abattoir 3.3.2 Abattoir

processor
3.7 Transport
3.4.1 Small scale 3.4.3 Wholesaler

processor
3.5.2 Import
3.4.3 Retailer
3.8 Local
consumers
Figure 2. Game meat supply chain in South Africa.

Legend:
Abattoir refers to an abattoirs approved for local and/or export purposes and where the slaughtering of
partial dressed carcasses is completed.
Farm abattoir refers to an abattoir not necessarily approved by the relevant authorities and where the
slaughtering of game carcasses is done for own consumption or the local market.
Large-scale processor refers to a processor approved by the relevant authorities for local and/or export
purposes and where value adding is done.
Small-scale processor refers to a processor not yet approved by the relevant authorities and where value
adding is done on a small scale for own consumption or the local market.
Wholesaler refers to an enterprise that trades in the local market with approved carcasses and/or value-
added products procured from approved abattoirs as well as large- and/or small-scale processors.
Retailer refers to an enterprise such as a local or supermarket butchery, biltong shop, meat selling point
(no processing), restaurant (including hotel), home industry, arts and crafts market, flea market, farmer
market, or any other locality where the raw and/or processed and/or cooked product is made available
directly to the consumer.

3.1 Primary producers (game farmers)
From a financial perspective it is widely recognised that game farming is the fastest growing
agricultural industry in South Africa (Patterson and Khosa, 2005) and increases by about
23% per annum in areas where there are no large predators (ABSA, 2003). Today there
is more wildlife in South Africa than 40 years ago (Bothma, 2004). Wild ungulates that
were objects of extermination prior to 1960 are now husbanded for a variety of purposes
(Carruthers, 2008). All of the sectors indicated in Figure 1 help the farmer ensure that wildlife
is a lucrative resource and a feasible land use option (Scriven and Eloff, 2003; Patterson
and Khosa, 2005). The South African wildlife industry is well established, with a range of
specialists including capture operators, wild-life transporting companies, wildlife marketers
or brokers, wild animal auctioneers, wildlife insurance brokers and wildlife management
consultants (Higginbottom and King, 2006).
Although the 65.7% of the game farmer respondents (n=134) indicated that they farmed
with game only, the remaining 34.3% indicated that they farmed with domestic animals and
game. In addition, 85.7% of the game farmer respondents indicated that their game was free
roaming and wild while the remaining 14.3% indicated that they farmed game intensively.
Regarding procurement strategies, 73.3% and 66.4% of the respondents (n=131) indicated that
they buy game at auctions and from other game farmers respectively for breeding purposes.
In addition, 24.4% of the respondents (n=131) indicated that they buy game at auctions and
from other farmers respectively for hunting purposes.
Kriek (2002) warns that the prevalence of internal and external parasites, hoofs and leg
problems due to malnutrition which in turn lead to fractures and nutritional deficiencies, e.g.
copper deficiency, may be of importance for game production when farmed intensively. In
addition, Bengis (2002) expressed concern regarding the dangers that game pose to domestic
livestock by acting as parasitic hosts or as a source of epizootic disease and also that foreign
animal diseases cycling in livestock may cross the interface and infect wildlife. From a
public health point of view, Paulsen and Smulders (2004) highlight that: (1) some diseases
are transmissible from animals to humans during handling (occupational hazard); (2) some
diseases are transmissible to consumers of meat and meat products; and (3) medical therapy
of meat animals could result in drug residues in meat and meat products. The control of
infectious pathogens that originate from wild animals have therefore become increasingly
important due to it having substantial impacts on human health, agricultural production,
wildlife based economies and wildlife conservation (Bengis et al., 2004). Table 1 is a summary
of diseases known to occur in game in South Africa.
Although certain diseases are identified by South Africa as notifiable, GlobalGAP which is
an international standard for ‘Good Agricultural Practices’ requires that a written Veterinary
Health Plan that supports optimal health of animals must be in place. This plan provides for
aspects such as the availability of the services of a veterinarian, disease prevention strategies,
vaccination protocols, endo and ecto-parasite control, identification and control of sick
animals, isolation facilities of sick or injured animals, control over storage and administering
of medication, maintenance of veterinary equipment and the keeping of records of animal
health interventions (GlobalGAP, 2010). In South Africa, 75.4% of the game farmer respondents

Table 1. Summary of diseases occurring in game in South Africa (Bengis, 2002; Bengis et al., 2001, 2004;
Godfroid, 2002; Keet et al., 2001; Michel et al., 2006; OIE, 2008, 2010).
Disease Species affected
African swine fever (OIE listed) Warthog (Phacochoerus africanus) associated with bites from the
eyeless tampan (Ornithodorus porcinus); domestic and free ranging
rustic pigs; and illegal translocated European wild boar in contact
with warthogs
African horse sickness (OIE listed) Zebra (Equus burchelli) associated with biting midge (Cullicoides
imicola)
Anthrax (OIE listed) Kudu (Tragelaphus strepsiceros)
Avian Influenza Farmed and wild ostriches (Struthio camelus)
Babesia White rhinoceros (Ceratotherium simum)
Bovine Malignant Catarrhal Fever (OIE Wildebeast (Connochaetes spp.) as maintenance host which infects
listed disease for wildebeest only) cattle
Bovine tuberculosis (Mycobacterium Baboon (Papio ursinus), buffalo (Syncerus caffer), cheetah (Acinonyx
bovis) (OIE listed) jubatus), hyena, impala (Aepyceros melampus), kudu, leopard
(Panthera pardus), lions (Panthera leo), meerkat (Suricate suricate)
and warthog
Brucellosis (OIE listed) Buffalo, eland (Taurotragus oryx), hippopotamus (Hippopotamus
amphibious), impala, sable antelope (Hippotragus niger niger),
waterbuck (Kobus ellipsiprymnus), zebra
Classical Swine Fever (OIE listed) Feral pigs and indigenous bush pigs (Potamochoerus larvatus)
Cyanobacterial intoxication Wildebeast, zebra, white rhino, buffalo, hippopotamus, giraffe
(Giraffa camelopardalis), warthog, lion and cheetahs
Feline Immunodeficiency virus infection Lions Kruger National Park (KNP)
Foot and Mouth Disease (OIE listed) African buffalo, bushbuck (Tragelaphus scriptus), giraffe, impala,
kudu, nyala, warthog and cattle
Hydatid disease (E. granulosis) OIE listed Necropsied lions
Newcastle disease (OIE listed) Farmed ostriches
Rabies (OIE listed) The viverid biotype were confirmed in common genets (Genetta
genetta), selous mongoose (Paracynictus selousi), slender mongoose
(Herpestes sanguinea), small spotted cats (Felis nigripes) and yellow
mongoose (Cynictus penicillata). The canid biotype were confirmed
in aardwolfs (Protoles cristata), African civet (Civettictus civetta),
bat-eared foxes (Otocyon megalotis), black backed jackals (Canis
mesomelas) and side striped jackals (Canis adustus)
Rift Valley ferver (OIE listed) Buffalo and gemsbok (Oryx gazella)
Theileriosis (Corridor disease) (OIE listed) Buffalo as carrier which infects cattle
Trypanosomiasis (OIE listed) Antelope, buffalo, elephant (Loxodonta africana), hippopotamus,
rhinoceros, wild porcine
Tuberculosis (OIE listed) (Mycobacterium Meerkat
tuberculosis)

(n=138) indicated that they do not have a written Veterinary Health Plan in place. In addition,
89.4% also indicated that they do not have a screening and improvement programme in place
for the control of zoonotic diseases. The absence of a written Veterinary Health Plan with the
necessary controls and records of disease control may result in the accidental spreading of
diseases to other animals, animal handlers and consumers of game meat.
3.1.1 Suppliers of feed
In nature, wild animals will move to better pasture, sometimes over long distances when
the availability of food is decreasing, usually during the dry seasons or drought (an example
of such migrations are that of the wildebeest in East Africa). However, with modern game
farming techniques, animals are fenced in which prevent them from leaving the fenced area.
Ninety percent of the game farmer respondents who indicated that game were free roaming
and wild also indicated that natural grazing was supplemented with feed during the dry
winter months in order to get optimum production. South Africa has a Fertilisers, Farm feeds,
Agricultural remedies and Stock remedies Act (Act 36 of 1974) that controls production and
import of farm feeds (Anonymous, 1974). Although not specifically designed for wild game,
GlobalGAP identify the following measures that game farmers may use as a guideline for the
control of animal feed (GlobalGAP, 2010):
• Feeds must only be procured from feed manufacturers or suppliers that are registered as
such by the relevant authorities.
• Labels of feeds must be checked and kept by the farmer as evidence of feed origin and
ingredient composition.
• Feed must be traceable to the manufacturer.
• Mixing protocols must be available when feed is mixed by the farmers themselves.
• A procedure should be in place to deal with residues of medicated feed.
• Records should be kept of the suppliers and the batch numbers received from the suppliers.
• Adequate measures should be taken to protect the feed against contamination, damage
and deterioration during storage. This includes pest and vermin control.
• Separate storage of medicated feeds in order to prevent cross contamination between
medicated and non-medicated feeds.
3.2 Harvesting and hunting
The majority (65.7%) of the hunter respondents (n=290) in the survey indicated that hunting
was part of the tradition of South Africans and was here to stay. Besides the estimated
200,000 local hunters, South Africa was visited by between 5,000 and 6,000 foreign hunters
during the 2003/2004 season (Patterson and Khosa, 2005). In South Africa, hunters can be
divided into professional hunters (PHs), sports or trophy hunters (SHs) and meat (biltong1)
hunters (MHs). In terms of the South African Firearms Control Amendment Act, 2006
(Act 28 of 2006), a professional hunter is ‘any person who supervises, escorts, offers to, or
agrees to supervise or escort a client, for reward in connection with the hunting of a wild
or exotic animal and who is authorised to do so in terms of any applicable provincial law’.
The same legislation also refers to an occasional hunter as ‘any person who, from time to
1 Biltong is a salted dried raw meat product widely eaten in South Africa and prepared from the muscle of the
antelope or cattle (Prior, 2008).

time, participates in hunting activities but who is not a member of an accredited hunting
association’ (Anonymous, 2006). Professional hunters are registered with the Professional
Hunters Association of South Africa (PHASA) after the completion of a comprehensive
training course at a professional hunting school.
3.2.1 Harvesting
Harvesting is a controlled process where compliance to specific legislation and standards is

compulsory (Van der Merwe, 2005). Such legislation includes the Animal Protection Act, 1962
(Act 71 of 1962) (Anonymous, 1962), the Meat Safety Act, 2000 (Act 40 of 2000) (Anonymous,
2000) and Veterinary Procedural Notices (VPNs) of the Department of Agriculture, Forestry
and Fisheries (Anonymous, 2008e). Harvesting can either take place during the day time by
making use of feeding point, boma (closure) or helicopter methods or during night time, by
using a spotlight from open vehicles in areas where less dense vegetation occurs (Hoffman,
2007). In accordance with South African rules the following matters should be in place before
and during the harvesting process of the game:
• Registration of a game farm for export status: an application is made to the relevant national
and provincial authorities to register the farm where harvesting is envisaged for export
status. This registration provides for official inspection and approval and registration of
farms; Requirements for residue monitoring as well as movement of partially dressed
game carcasses from a registered farm to an abattoir. Prior to harvesting, officials from
the relevant authorities must obtain a duly signed health attestation regarding the animal
health status on the farm from the regional State Veterinarian (Anonymous, 2008a). This
ensures that there is or was no recent disease outbreak on the farm or the direct vicinity
of the farm.
• Harvesting teams: the team leader must ensure that the hunters and other staff have
adequate competencies in hunting and slaughter techniques, that they are registered as
such by the authorities and that they have adequate knowledge of legislative requirement
(Anonymous, 2008b). At least one of the team members must be qualified as a Game
Meat Examiner (GME) or a Game Meat Inspector (GMI). In South Africa a person can
qualify as a GME after the successful completion of a six month competency based course
while a GMI must be in possession of at least an appropriate three year tertiary training
qualification.
• Temporary slaughter depots: the harvesting team must provide temporary slaughter
depots where field slaughter is conducted (Figure 2, item 3.2). The depots must provide for
(1) hard rolled floor surfaces covered with canvas or plastic to control dust; (2) hanging
facilities for the dressing of carcasses and handling of viscera and high enough to prevent
the head or neck of the carcass coming in contact with the ground; (3) meat inspection
facilities for carcasses and viscera; (4) separate closable and washable containers for the
storage of red offal, rough offal, inedible material and condemned material; (5) sterilizers
for knives and other slaughter equipment; (6) sufficient potable water with hot running
water at 40 °C or with an acceptable food grade approved disinfectant added; (7) adequate
illumination for night slaughter (220 Lux for dressing and 540 Lux at the inspection
point); (8) hand wash and disposable hand drying facilities with a supply of food grade
approved liquid bactericidal soap; and (9) mobile thermograph equipped chilling facilities
(preferably refrigerated trucks) with the capacity to reduce the carcass temperature to

below 7 °C within 24 hours for storage and transport of slaughtered carcasses and process.
The accumulation of blood, waste, dust and mud below the frame and surrounding areas
must be prevented throughout the slaughter process (Anonymous, 2008b). Samples of
the water are taken at least one week before the harvest in order to determine the quality
of the water.
• Hygiene management system (HMS): harvesting teams must have in place a HMS that
provides for the control of ante mortem inspection, slaughter and dressing, personal
hygiene of workers, medical fitness of workers, maintenance of sterilizers (chemical or
high temperature sterilisers), availability of liquid soap and soap dispensers, toilet paper,
and disposable towels, sanitation and continuous cleaning, availability and safety of water,
waste disposal, including condemned material and continuous temperature control of
the chiller vehicle (Anonymous, 2008b).
• Pre-harvesting inspection: the leader must ensure that the game to be harvested was
not treated with veterinary medicinal drugs before harvesting or that all withdrawal
periods for such veterinary medicines were adhered to. Prior to the start of harvesting,
the slaughter depots and harvesting vehicles must be inspected by the GME or GMI to
determine compliance with the hygiene requirements. In addition the hunting team must
also have available for inspection by the GME or GMI the registration certificate(s) of the
farms and for all hunters on the team, health attestation of the farm issued by the relevant
authorities, harvesting programme, health certificates of hunter(s) as well as of assistants
including copies of their identification documents, checklist for harvesting inspection,
certificate of origin and proof of qualification the hunter is qualified to do post mortem
inspections (Anonymous, 2008b,c).
• Shooting/killing: it is the responsibility of the hunter to ensure that the animals harvested
has a normal healthy active appearance and that no animal with visible signs of injury or
disease are hunted. If killed, hunters must mark all suspect animals and provide relevant
information to the meat inspectors at the slaughter depot. The use of Professional Hunters
is preferred as the meat derived from harvested game is mostly exported and only head
and neck shots are used. Game killed with thoracic shots must be subjected to veterinary
approval while carcasses with abdominal shots must be condemned for export purposes.
Carcasses not approved for export purposes may however be considered for domestic use
when it is fit for human consumption (Anonymous, 2008b,c).
• Bleeding: is done by means of severing the jugular vein and carotid artery on either side of
the neck (throat slitting). As with domesticated animals, bleeding of game animals should
be done within as short of time as possible after the kill. Considering that bleeding in the
hunting field is not always that easy because of the terrain and the time lapse to get to the
hunted animal, South African standards allows a maximum of 10 minutes after shooting
(Anonymous, 2008b). Observations during harvesting however showed that bleeding
takes place within 2-5 minutes after shooting, which is shorter than the prescribed time.
South African standards require that small animals be bled in a hanging position; medium
animals on a ramp at 20-30°, or in a hanging (elevated) position and large animals in a
lying position. The bleeding knife used must be cleaned and sterilised by using water at
82 °C or an approved food grade chemical method. The provision of an adequate number
of knives and the use of a two knife system is recommended in order to ensure the effective
sterilisation of the knife not in use (Anonymous, 2008b). It is also important to ensure
that the contact time is adequate for effective sanitation of the knives.

• Transport of harvested game: South African standards require that harvested game must
be transported to a game depot within two hours after being bled. To shorten the time it
is suggested that carcasses are off-loaded at the slaughter depot(s) as soon as they are in
close vicinity thereof, even if the load is not full. Observations made during harvesting
showed that on average, carcasses are transported to the depot within 45 minutes post
mortem, which is shorter than the allowed two hours (Anonymous, 2008b). Care must be
taken not to contaminate the neck slit area when transporting the carcasses. The vehicles
used to transport harvested game carcasses from the point of kill to the farm abattoir
(field slaughter depot) must be equipped with a hanging frame and/or a hoist and a ramp
frame with a 20° to 30° slope to bleed carcasses. In addition vehicles must be equipped
with facilities where bleeding knives can be cleaned and sterilised with water at 82 °C or by
means of food grade chemical sterilisation; hand wash facility with potable running water
and soap, and artificial light where game is bled at night with a minimum light intensity of
220 Lux. The structure of the frames, sterilisation equipment and had wash facilities must
be constructed of a non-toxic material that is smooth surfaced, non-absorbent, resistant
to impact and durable and easy to clean (Anonymous, 2008b).
• Field slaughter: is done at the temporary slaughter depot by transferring the bled animals
from the harvesting vehicle onto a hanging frame where the heads, feet, lactating udders,
scrotum and testicles are removed (leaving the Lnn. inguinalis superficialis) and carcasses
are eviscerated in a hanging position. The hide or skin is however not removed and
acts as protection against environmental contamination during the slaughter process.
Evisceration is normally done at the slaughter depot, but as harvesting can take place
any time of the year, bloating may occur in the hotter months of the year. If bloating
occurs, the carcass must be brought to the depot sooner or the removal of the green offal
(paunch and intestines) can as an emergency measure be removed by the hunter in the
field within 30 minutes of being bled (Anonymous, 2008b). In such cases the hunter who
is also a qualified GME or GMI must inspect the green offal during evisceration in the
field. As with slaughter of domesticated animals in commercial abattoirs, the carcasses
and the corresponding viscera must be identifiable for meat inspection purposes at the
depot and abattoir. Throughout the field slaughter extreme measures must be taken to
prevent contact of the exposed meat with soiled equipment, platforms, slaughter frames,
ground or floor as well as the outer surface of the skin or hide. No cutting of the Rectum,
small intestines, oesophagus, bladder and uterus must be allowed.
• Preliminary post mortem meat inspection: carcasses and the corresponding viscera must
be inspected by qualified GMEs or GMIs. A comprehensive post mortem meat inspection
must be done on the partially dressed carcass as well as all heads and feet, oesophagus,
trachea, lungs, spleen, heart, kidneys (if removed), diaphragm, liver, mediastinum,
abdominal organs. The partially dressed carcasses are inspected for thoracic shots resulting
in gross contamination of the thoracic cavity with blood and/or bone splinters or gut
shots resulting in contamination of abdominal cavity with ingesta; signs that an animal
was wounded or required more than one shot to kill; and excessive contamination of the
abdominal cavity with ingesta, soil, grass, mud or any other contaminant where game
animals were eviscerated in the field employing poor evisceration techniques. Although
such animals are not used for export it was found during observations that depending
on the reason for the rejection of the carcass and, where possible, some carcasses such as
heavily bruised carcasses remained on the farm for consumption by farm staff or further

processing of the parts of the carcasses still fit for human consumption (Anonymous,
2008c).
• Washing of partially dressed carcasses: in order to minimise contamination and to keep
the carcass as dry as possible, no partially dressed carcass may be washed and accidental
soiling/ contamination must be cut off (Anonymous, 2008c).
• Traceability: in order to ensure traceability to the abattoir, carcasses and corresponding
pluck (heart, lungs and liver) must be tagged, each set with its own unique identification
code. A list of the codes is attached to a Certificate of Origin that accompanies the
refrigerated transport vehicle (Anonymous, 2008c).
• Chilling: after primary meat inspection the carcasses must be left to air dry. South African
rules require however that inspected carcasses must be loaded into refrigerator trucks
within 4 hours of inspection during summer months, which can be extended to 12 hours
with an ambient temperature of 12 °C or less (Anonymous, 2008c). It was observed that
on average the carcasses were loading into the refrigerated vehicles within three hours
after inspection. Refrigerator vehicles must be able to chill the partially dressed carcasses
to a deep bone temperature of below 7 °C within 24 hours (Anonymous, 2008b).
• Offal handling: the red offal must accompany the partially dressed carcasses in separate
containers to the abattoir where final slaughter and inspection is conducted. As already
indicated these must be suitably identified and must correlate with the carcass. The
remaining offal after approval may be used by local consumers (Anonymous, 2008c).
• Transport: vehicles transporting partially dressed game carcasses and red offal must
comply with regulations pertaining to vehicles transporting meat in terms of the design
of the walls, floor and roof, i.e. it must be smooth, non-absorbent and washable. When
all partially dressed carcasses are loaded, the GME or GMI must seal the truck with an
official seal and record the unique seal number on the Certificate of Origin. The transport
vehicle must be able to maintain a deep bone temperature of between -1 °C and 7 °C until
offloading. A thermograph recording the temperature continuously must be available and
the recording must provide for accurate actual time and temperature analysis, covering
all phases of loading during the slaughter process and transport. Carcasses must be hung
away from the floor and from each other in such a way as to ensure optimal airflow within
the chiller space. The load must be accompanied by the health attestation, the Certificate
of origin, the checklist for harvesting inspection and the inspection report of the GME
or GMI (Anonymous, 2008c).
• Waste handling: lactating udders, reproductive organs and any organ not utilised
commercially must be handled as condemned material and placed in appropriate
containers. Condemned material must also be placed in the same containers and disposed
of in an appropriate manner (Anonymous, 2008b,c).
• Monitoring for pharmaceutical substance residues: sampling of liver, fat, kidney, muscle
tissue, blood serum and urine must be carried over a period of time and spreaded over the
whole hunting season. The sampling programme must make provision for harvesting that
may take place throughout the year in some areas, specific substances that are administered
only in particular, the use of currently unknown substances and the presence of diseases
in particular regions (Anonymous, 2009).

3.2.2 Hunting
Hunting refers to the act of chasing and killing wild animals for sport or food and is mostly
an uncontrolled process, not necessarily following prescribed legislation. It sometimes takes
place under the guidance of a professional hunter and is conducted by hunters that are not
professional hunters (Van der Merwe, 2005). Hunting normally takes place in areas where
wildlife viewing or photographic safaris are not possible (Barnett and Patterson, 2006). The
following types of hunting are identified in South Africa (Humavindu and Barnes, 2003;
Patterson and Khosa, 2005; Carruthers, 2008):
Trophy hunting: the hunter (national and foreign) is a client to a hunting concession owner
and hunts purely for sport reasons with the objective of keeping some part of the animal as
proof of the hunt. Once the hunter has taken the trophy, usually the horns, skull and/or skin,
the left over meat is either used as bait for a subsequent hunt, or is given to local staff or local
communities, although the hunters may request some for his/her own meals. Trophy hunting
has became more and more popular and was the initial stimulus to develop wildlife ranching
as a major economic force which generated an estimated income of R417 million for game
ranchers in 2005 (PHASA, 2006). The trophy hunter’s main objective is an undamaged trophy
and not the health or hygiene of the meat (Van der Merwe, 2005). Game farmer respondents
(n=114) indicated that on average 34.2% of meat obtained from trophy hunted animals was
taken by the hunter, while 32.5% indicated that the meat remained on the farm, 17.5% said
that it was taken by the professional hunter that accompanied the hunter during the hunt
and 15.8% stated that the meat was sold to interested persons.
Biltong/meat hunting: the hunters are almost exclusively South African nationals who
combine the experience if hunting with the desire for wildlife meat. The skin, horns or
any other animal parts are seldom kept by the hunter as a trophy and sometimes hunting
concession owners generate an income from these parts. The biltong hunter’s objective is
the thrill of the hunt and the biltong is a bonus (Van der Merwe, 2005). Van der Merwe
and Saayman (2008) indicated that in 2007 the top five species hunted for biltong were
springbok (Antidorcus marsupialis), impala (Aepyceros melampus), blesbok (Damaliscus
dorcas philipsi), kudu (Tragelaphus stepsiceros), warthog (Phachochoerus africanus) and blue
wildebeast (Connochaetes gnou).
Traditional hunting: traditional hunting refers to those whose hunting activity is aimed at
provision of food, the satisfaction of cultural practices and/or the source of medicinal material
and to test the speed of hunting dogs.
Subsistence hunting: this is where wildlife is hunted for its meat using a wide variety of
methods and does not take place within any legal framework. This method is also frequently
referred to as bush meat; although the latter may also include the illegal hunting of wild
animals with the intention of selling the meat in an informal (and thus illegal) manner.
Very little of the rules that apply to harvesting are applied during hunting. Hunters may or
not have some of the basic equipment that will assist them with the hunt such as a vehicle
equipped with a hanging frame and a spotlight, a container with water and slaughter knife

to do the bleeding and the removal of the green offal (paunch and intestines). Bleeding and
removal of the green offal mostly takes place on the hunting ground and is normally done
by the hunter or an assistant that accompanies the hunter during the hunt. From there the
carcass is transported on an open vehicle to the farm abattoir or transported to the hunter’s
residence (Figure 2, Item 3.2.1) where the slaughter is completed and sometimes the carcass
is further processed into various products. Normally no meat inspection is conducted on the
carcass or the viscera. However, the survey conducted indicated that meat and meat products
derived from wild game animals is highest in demand by butcheries (43.3%) and consumers
(90%) during winter months, which traditionally is the hunting season of the year. Although
the majority of the hunter respondents in the survey indicated that they do not have adequate
knowledge of legislation controlling meat hygiene and safety, 81.1% of the hunters (n=280)
indicated their willingness to attend information sessions in order to obtain more knowledge.
It is therefore important to ensure that all hunters receive relevant training to ensure that
the meat and meat products derived from hunted wild animals are safe for consumption.
3.3 Slaughter
In South Africa, the Sub-directorate for Veterinary Public Health of the Department of
Agriculture, Forestry and Fisheries (DAFF), is responsible for the slaughter of animals within
approved abattoirs while the Sub-directorate Import/Export are responsible for the import
and export of game meat. General Law enforcement is conducted by the veterinary public
health units of the provincial government in their areas of jurisdiction.
3.3.1 Farm abattoirs
A farm abattoir is a slaughter facility that is not necessarily approved by the relevant authorities
and where slaughter of game carcasses is done for own consumption or for the local market.
Although the majority (91.6%) of the game farmer respondents in the survey indicated that
they offered a slaughter service to hunters, the locations and the number of these abattoirs are
unknown and therefore none or little control can be executed. No inspection of the hunted
carcasses took place in the hunting field. In addition the carcasses of those game species
wrongly shot during harvesting as well as carcasses with excessive damage or bruising and
therefore not suitable for export purposes may end up in the farm abattoir for final slaughter.
Normally no final meat inspection will take place at these farm abattoirs in order to determine
the fitness thereof for human consumption. In cases where a farm abattoir is not available
on the farm where the hunting take place, the carcasses of the hunted game is normally
transported to another nearby farm abattoir or another abattoir registered for game slaughter
(Figure 2, item 3.3.2) for final slaughter or the hunters residence (Figure 2, item 3.2.1) for final
slaughter and further processing. The majority (80%) of hunter respondents indicated that
they conduct the final slaughter and processing of the hunted game on the farm where the
game was hunted. Meat for own consumption is exempted from inspection, but if meat is
sold to an outlet or directly to the public, hunters have to meet the requirements of the Meat
Safety Act, 2000 (Act 40 of 2000) (Anonymous, 2000; Patterson and Khosa, 2005) as well as
the ‘General hygiene requirements for food premises and the transport of food’ promulgated
under the National Health Act (Act 61 of 2003). It was evident from the survey that game
meat reaches the consumer through the formal supply chain. Hunters (n=279) indicated that

they already market their hunted game meat prior to the hunt to game farmers for own use
or further processing (7.9%), schools or other charity organisations (6.5%), family or friends
(39.4%), wholesalers (3.4%) and retailers (24.8%) such as local and supermarket butcheries,
biltong shops and restaurants. Forty percent of the game farmer respondents indicated that
they procured game carcasses from hunters to process game meat products on the farm with
the specific purpose of selling the meat or processed meat products to the retail (Figure 2,
item 3.4.3) and wholesale markets (Figure 2, item 3.4.3) or directly to the consumer (Figure 2,
item 3.8). However, in terms of fitness for consumption, 75% of hunter respondents indicated
that the meat is not inspected by anyone, while the others indicated that the carcasses and
offal are inspected by qualified game meat examiners (5%), hunters themselves (10%), the
farmer (12%), or another hunter (3%). However, none of the latter three parties have received
any training in game meat examination. This situation clearly states a gap in control in the
meat supply chain.
3.3.2 Abattoir (game meat establishment)
The abattoir or as it is also referred to, the game meat establishment is defined in terms of the
Meat Safety Act, 2000 (Act 40 of 2000) (Anonymous, 2000) as ‘a slaughter facility in respect
of which a registration certificate has been issued and in respect of which a grading has been
determined’. In addition, separate regulations to manage the registration process, slaughter
processes and hygiene management have been promulgated in terms of the said Act for red
meat (bovine, ovine, porcine and equine), poultry meat and ostrich meat. Regarding game
meat, draft Game Meat Regulations were published for comments during 2004 under the same
Act but has still not been promulgated since an amendment needs to be made to the said Act,
which currently prohibits the receiving and slaughter of dead animals in an abattoir. However,
due to the administrative and prolonged nature of the amendment process, 27 abattoirs are
already approved for the slaughter of game in South Africa (Anonymous, 2008d). Some of
these abattoirs also slaughter other domesticated red meat animal species such as bovines,
ovine, porcine and ostrich. It is the responsibility of the abattoir owners or managers to ensure
that the abattoir is maintained and the slaughter process is conducted in accordance with the
prescribed standards. In order to achieve this it is expected of abattoir owners or managers
to implement a documented Hygiene Management System consisting of (Anonymous, 2004):
• schematic plan of the abattoir;
• flow diagram of the slaughter process;
• identification of potential biological, chemical or physical hazards and the prevention
thereof;
• hygiene management programmes relevant to game slaughter including slaughter and
dressing, meat inspection, personal hygiene and medical fitness of workers, temperature
of water in sterilizers and maintenance of sterilizers, availability of liquid soap and soap
dispensers, toilet paper, and disposable towels, sanitation and continuous cleaning,
availability and quality of water, vermin control, waste disposal, contact wrapping and
packing materials, maintenance, thermo control and visitor control.
Although it is also expected of abattoir owners or managers to conduct a regular self

assessment based on a prescribed Hygiene Assessment System (HAS), veterinary public

health officials employed by the respective provinces also evaluate the abattoirs on a regular
basis against the same HAS (Anonymous, 2000, 2004).
Partially dressed carcasses and their corresponding plucks are delivered to abattoirs. If the
carcasses originate from a harvest, the seal is checked against the Certificate of Origin and
ensured that it is still intact and the number of carcasses concurs with the prescribed register
that accompanied the vehicle. The carcasses are offloaded and stored in holding chillers before
slaughter is completed by removal of the hides and skins and comprehensive final primary and
secondary meat inspections are conducted on the carcasses and the accompanying viscera. The
approved carcasses are normally deboned or processed into primal cuts, chilled or frozen and
stored until dispatched for export (Figure 2, item 3.5.1) or to the local markets, i.e. large-scale
processors (Figure 2, item 3.4.2), retailer and wholesalers (Figure 2, item 3.4.3) or directly to
the local consumer (Figure 2, item 3.8). The results of the ante mortem inspection in the field,
primary meat inspection and secondary meat inspection must be recorded and where zoonotic
and notifiable diseases are diagnosed, the local state veterinarian must be notified.
3.4 Processing, wholesale and retail markets and export
Something that causes some difficulty in control of game meat (and for that matter all other
meats) in South Africa is that the control of the meat after the gate of the farm abattoir (Figure
2, item 3.3.1) or the abattoir premises (Figure 2, item 3.3.2) falls under the metropolitan and
district municipal health authorities jurisdiction and no longer with the relevant veterinary
public health authorities. There is very little liaison and cooperation between the veterinary
public health authorities, the municipal health authorities and the abattoirs. The mandate and
role of municipalities in food control lies within the Foodstuffs, Cosmetics and Disinfectants
Act, 1972 (Act 54 of 1972) (Anonymous, 1972) and the National Health Act, 2003 (Act 61 of
2003) (Anonymous, 2003). The Foodstuffs, Cosmetics and Disinfectants Act, 1972 (Act 54 of
1972) deals with matters regarding the sale, manufacture and importation of foodstuffs while
the latter Act deals with hygiene requirements for food premises and the transport of food.
Typical hygiene requirements include aspects such as the registration of the premises and the
issuing of Certificates of Acceptability; handling and transportation of food; structural and
design requirements for food premises; facilities, containers and equipment; display, storage
and temperature control of food as well as food handlers and protective clothing. Although it
is the primary responsibility of the enterprise owners or managers to ensure compliance with
the legislation, the control over the implementation and maintenance of the requirements
are done by authorised Environmental Health Practitioners (EHPs) and in selected cases by
veterinarians (Basson, 2006). Visiting EHPs are also responsible for sampling and analysis
of foodstuffs; health education of food processors, handlers and consumers and advising
on legal requirements (Basson, 2006). The survey has however shown that 7% of local and
supermarket butcheries, meat processing plants and restaurants, 14% of wholesalers and
16% of biltong shops are not in possession of Certificates of Acceptability. By not issuing the
required Certificates of Acceptability, municipalities are not aware of food premises or they
are ignoring their responsibilities regarding this matter.
In addition to hygiene and processing requirements stated by legislation it is imperative for

all processors to establish, implement and maintain a food safety management system i.e. a

Hazard Analysis Critical Control Point (HACCP) system or The Food Safety Management
System of the International Standards Organization (ISO, 2005). The purpose of the mentioned
system is to establish policies and procedures whereby food hygiene, safety and quality can
be managed. In terms of the requirements of such as system a hazard analysis of all processes
and products are conducted and the necessary controls are implemented to prevent and
control identified hazards.
3.4.1 Small-scale processors
The term ‘small-scale processors’ refers to processors not yet approved by the relevant
authorities and where value adding is done on a small scale for own consumption or the local
market. Small-scale processing is often done on game farms in a separate room adjacent to the
slaughter area. Seventy percent of the game farmer respondents indicated that they processed
game meat products on the farm themselves and that the top five products processed were
biltong (50%), dried sausage2 (30%), salami (10%), fresh cuts (5%) and fresh sausage (5%). This
confirms the finding of Hoffman et al. (2004) that there is still a demand for the processing
of biltong (including dried sausage) as a delicacy amongst South African consumers. Other
products include mince, hamburger patties, and cabanossi sausages. Although game meat
is not new to the South African consumer, it is no surprise that processors experiment with
alternative processed products in order to satisfy the growing need for lean meat (Hoffman
and Wiklund, 2006). According to the survey conducted most (49.4%) of the game meat
processed on the farm remains on the farm for own home use, use in game lodges; use
as rations for farm workers; and selling to hunters and interested persons or the public.
Approximately 17% is donated to family, friends or charity and 31.9% of the processed meat
processed on the farm reaches the consumer (Figure 2, item 3.8) through local retail and
wholesale markets (Figure 2, item 3.4.3).
Currently, neither veterinary public health nor municipal health authorities know the location
and number of farm processing facilities and these are therefore not yet approved in terms of
the applicable legislation referred to above. Another complicating factor is that the amount
of game meat received, processed and sold in municipal areas of jurisdiction is unknown to
the municipal health authorities.
3.4.2 Large-scale processors and exporting
Large-scale processors are normally located in urban areas and this definition refers to
processors approved by the authorities for local and/or export purposes and where value
adding is done and the final products are distributed to the wholesale (Figure 2, item 3.4.3) and
retail markets (Figure 2, item 3.4.3) from where the local consumers purchase the products.
2 South African dried sausage (droëwors) is a ready-to-eat dried seasoned and intermediary moisture meat
product which can be distinguished from European types of dried sausages in that it is not cured nor fermented,
but preservation is obtained by an artificial drop in pH and subsequent drop in water activity (Burnham et al.,
2008; Osthoff et al., 2002).

3.4.3 Wholesalers and retailers
As previously indicated, game meat and game meat products also enter the supply chain at
the wholesale and retail points. This was confirmed when butchery respondents in the survey
indicated 64.5% of the game meat processed and sold by butcheries is obtained from hunters
(including own hunting). In addition, 52.2% confirmed that that hunters offer game meat
for sale to them prior to hunting, while 36.2% indicated that they deliberately order or buy
game meat from hunters for the purpose of processing and sale in their butchery. Butchery
respondents also indicated that only 63.6% of the game meat received by their butcheries
is processed on behalf of leisure hunters for their own use while the remaining 36.4% of
the meat is processed and sold directly to the consumer from the butchery. Carcasses are
mainly (63.4%) received without the hide or skin which complicates species identification.
On the other hand, the remaining 36.6% that are received with the hide on requires separate
chilled storage facilities to prevent cross-contamination of other carcasses. It must also be
remembered that the carcasses obtained from hunters are not inspected by a qualified PME or
PMI. The absence of the head, feet and viscera at this point also prevent complete inspection
of the carcass and its entrails as required by the legislation. Despite of all these complicating
factors, only two of the municipality respondents indicated that they have policies specifically
aimed at the control of game meat entering their area of jurisdiction.
3.5 Export and Import of game meat
Import and export of game meat is an important contributor to the economy, but also forms
part of the larger goal of sustainable food security in South Africa.
3.5.1 Export
Approximately 450 tons of game meat are exported annually (mainly to Europe) with a
value of about R15 million (Du Toit, 2007). In South Africa there are five game handing
establishments approved for export purposes by the European Union (EC, 2008). Two of them
are also approved as cutting plants. These facilities as well as the export of the game meat are
controlled by the Sub-directorate: Import/Export within the Directorate: Veterinary Health
of the Department of Agriculture, Forestry and Fisheries. It is expected of these facilities
to comply with the requirements of the Meat Safety Act, 2000 (Act 40 of 2000) as well as a
series of Veterinary Procedural Notices (VPNs) prescribed by the same sub-directorate. The
VPNs prescribe procedures in line with European Union standards and deal with controlling
of animal diseases on the farm; hunters used for harvesting; ante and post mortem meat
inspection and hygiene control at point of harvest and game meat establishments; control of
meat; residue monitoring; and law enforcement (Anonymous, 2008e). These establishments
also have full time veterinarians and meat inspectors on site that oversee the compliance
with the requirements.
3.5.2 Import
Game meat is imported from approved cutting plants in countries such as Australia, Namibia
and New Zealand, mainly for commercial purposes, but also by hunters who have hunted

in neighboring and other countries. The meat derived from the latter may be for the hunters
own use but also for commercial purposes. Although policies regarding the import of meat
are determined by the Import/Export unit within the Directorate: Veterinary Services of
the Department of Agriculture, Forestry and Fisheries, the Sub-directorate: Port of Entry
Point Control of the same department administers these policies. Control policies consider
the following:
• Restriction of the mass of meat for own consumption. South Africa allows a maximum
of 250 kg of meat per individual for own consumption. The import of meat exceeding
250 kg per importer is regarded as import for commercial purposes (Anonymous, 2008f).
• Issuing of veterinary import permits and veterinary health certificates by the relevant
veterinary authorities. Specific conditions regarding the import may also be stipulated
in the permit i.e. temperature requirements during transport and proof that the meat is
coming from a Foot and Mouth free area (Anonymous, 2008f).
• Animal health status at the point of origin of the meat (Anonymous, 2008f). In South
Africa this matter is evaluated and approved by the Directorate: Veterinary Services of
the Department of Agriculture, Forestry and Fisheries.
• Approval of game meat processing plants in various countries: these are visited and when
found to be complying with the relevant requirements they are registered as approved
suppliers. Currently, South Africa has plants approved as suppliers in Australia, Namibia
and New Zealand (Anonymous, 2008g).
• Placement of governmental officials with relevant knowledge and experience at air, land
and sea ports of entry to ensure that the meat is imported in accordance with requirements
of the import permit (Anonymous, 2008f). In South Africa, inspectorate of the Sub-
directorate: Port of Entry Point Control is responsible for this function. On approval of
meat for own consumption, the meat is released into the hands of the owner who is the
consumer (Figure 2, item 3.8).
• Final inspection and approval of consignments meant for the commercial market. In
South Africa, the meat is finally inspected and approved by a state veterinarian at a
designated end-point before it is released (Anonymous, 2008g). Only thereafter, the
meat may be distributed to the large-scale processor (Figure 2, item 3.4.2), the wholesaler
(Figure 2, item 3.4.3) and the retailer (Figure 2, item 3.4.3).
3.6 Suppliers of processing materials
Suppliers of any processing materials such as ingredients, additives and packaging material are
primarily responsible for supplying materials that are safe and of a high quality. The import
of any processing materials is regulated under the Foodstuffs, Cosmetics and Disinfectants
Act, 1972 (Act 54 of 1972) (Anonymous, 1972). Law enforcement at establishments where
manufacturing take place is done by Environmental Health Practitioners employed by the
municipal health departments. Composite products such as pre-mixed spice mixes must
indicate the ingredients on the label in order for the user to identify ingredients such allergens,
stabilisers and colorants. This allows the processor to determine compliance with the relevant
and local legislation.

3.7 Transport
Game meat is transported from the point of kill throughout the supply chain to the customer.
Even though standards and regulations exist, the control of hygiene and safety practices
through this part of the supply chain is often neglected and may have a negative impact on
the safety and quality of the meat. Table 2 provides information on controls that can to be
taken during transport through the supply chain.
Table 2. Description of transport methods in the supply chain as observed during the study.
Transport Controls
Point of kill to field slaughter depot Protection of exposed neck bleeding slit and abdomen in cases where
(Figure 2, item 3.2) or farm abattoir the green offal is removed in the field.
(Figure 2, item 3.3.1) on an open pick-up Prevent neck or carcasses coming in direct contact with the ground.
vehicle. Keep the time from kill to delivery at slaughter depot or farm
abattoir as short as possible.
Field slaughter depot (Figure 2, item 3.2) The thoracic cavity is open and its viscera, the head and feet are
to farm abattoir (Figure 2, item 3.3.1) on removed and it is therefore necessary to protect the exposed parts
an open pick-up vehicle against contamination during transport.
Field slaughter depot (Figure 2, item Proper design of the internal structure of the refrigerated vehicles to
3.2) to abattoir (Figure 2, item 3.3.2) in enhance cleaning and prevent contamination.
refrigerated vehicles. Installation of a refrigeration unit capable of maintaining the
temperature below 7 °C.
Equip vehicle with thermometer couplings linked to a thermo
control device that will monitor the temperature continuously.
Sealing of refrigerating unit doors to prevent unnecessary opening of
the doors during transport.
Non-edible or rejected products may not be transported in the same
compartment as carcasses.
Measure and record carcass temperature during loading and off-
loading.
Farm abattoir (Figure 2, item 3.3.1) to If the carcass is transported with the hide or skin on, the carcass
local consumer (Figure 2, item 3.8) for must be protected against contamination.
own use in the consumer’s (including If the hide or skin is removed, the meat must be protected against
hunter’s) own vehicle(s). contamination by packing in suitable clean containers.
If the meat was chilled and is transported over a short time
(maximum one hour), the meat can be transported in cooler boxes
or something similar that will maintain the temperature.
Transport over longer times must be refrigerated.

Table 2. Continued.
Transport Controls
Farm abattoir (Figure 2, item 3.3.1) to Proper design of the internal structure of the refrigerated vehicles to
small scale processors (Figure 2, item enhance cleaning and prevent contamination.
3.4.1) or wholesaler and retailer markets Installation of a refrigeration unit capable of maintaining the
(Figure 2, item 3.4.3) in refrigerated temperature below 7 °C when fresh or below -18 °C when frozen.
vehicles. Equip vehicle with thermometer couplings linked to a thermo
control device that will monitor the temperature continuously.
Sealing of refrigerating unit doors to prevent unnecessary opening of
the doors during transport.
Non-edible or rejected products may not be transported in the same
compartment as carcasses.
Measure and record carcass temperature during loading and off-
loading.
Abattoir (Figure 2, item 3.3.2) to large The same precautions as with the previous item will apply.
scale processor (Figure 2, item 3.4.2) or
wholesaler and retailer (Figure 2, item
3.4.3) in refrigerated vehicles.
Large scale processor (Figure 2, item 3.4.2) The same precautions as with the previous item will apply.
to point of export (Figure 2, item 3.5.1)
in refrigerated vehicles.
Point of import (Figure 2, item 3.5.2) to The same precautions as with the previous item will apply.
large scale processor (Figure 2, item
3.4.2) or wholesaler and retailer (Figure
2, item 3.4.3).
Point of import (Figure 2, item 3.5.2) The meat must be packed in suitable clean containers to protect it
to consumer (Figure 2, item 3.8) for against contamination.
own use in the consumer’s (including Meat must be chilled or frozen and maintained at temperatures
hunter’s) own vehicle(s) and is normally below 7 °C when fresh or below -18 °C when frozen.
without the hide or skin.
Point of purchase by consumer to Consumer and retailer education regarding the maintenance of the
consumer’s home mostly in consumer’s cold chain and how it can be maintained when shopping.
own vehicle with no refrigeration or Encouraging of retailers to make cooler bags available for sale to
facility to maintain the temperature. customers.
3.8 Consumers
Consumer confidence in the quality (including safety) of their food supply depends in part
on their perception as to the effectiveness of food control measures (CAC, 1995). Although
consumers consider game meat consumption as a healthier alternative to red meat (Hoffman
and Wiklund, 2006), they also consider aspects such as quality, i.e. tenderness, leanness,
juiciness; meat safety i.e. the presence of antibiotics, chemical residues and growth hormones;

and animal welfare, i.e. health status and stress that the animals were subjected to (Krystallis
et al., 2006; Blokhuis et al., 2008).
It is however often the customers themselves that cause undesired changes in the game meat
through aspects such as temperature abuse, unhygienic handling and poor preparation
practices. Although consumer education is an enormous challenge, stakeholders in the
game meat supply chain should take responsibility to educate consumers on proper handling
practices. The offering of programmes such as the ‘Five Keys to safer food’ of the World Health
Organization (WHO, 2006) at schools, universities and other public forums may assist to
overcome this problem. The five keys relate to (1) Keeping clean; (2) separation of raw and
cooked food; (3) Cooking food thoroughly; (4) Keeping of food at safe temperatures and (5)
the use of safe water and raw materials.
4. Conclusions
South African game meat and game meat products obtained from harvesting or hunting
as well as imports and exports are reaching both the local and the international consumer
through the game meat supply chain described in this chapter. In order to assist policy makers,
law enforcers and the game meat industry, essential foods safety control points were described
and are summarised in Table 3.
The establishment and implementation of proper and appropriate policies, legislation and
programmes by the respective stakeholders in the supply chain as appropriate to what they
are responsible for will prevent potential food safety breakdowns in the supply chain for
game meat.
The essential food safety management points described applies only to the hunting of larger
game (antelope) such as impala, springbok, kudu, zebra, eland, etc. and does not include
the supply chain for (1) bird hunting, i.e. guinea fowls, partridge, etc. or (2) bushmeat that
is obtained through informal and frequently illegal hunting of other wildlife such ascertain
rodents, reptiles, porcupine and birds.
More research is needed on the supply chain and the appropriate essential food safety
management points for the harvesting and hunting of birds and bushmeat. Bush meat is often
also associated with traditional practices of certain communities and a better understanding
of these traditional practices may assist with the establishment of food safety management
points. Although bird hunting is not done on large scale in South Africa and is mostly used
by the hunters and direct communities, it has the potential to also reach the formal supply
chain. Aspects that may impact on the hygiene and safety as well as the general quality of

Table 3. Essential food safety management points for game meat.
On the game farm After the game farm
Compliance with national and international legislation. Compliance with national and international legislation.
Implementation of an animal health plan. Transport control throughout the supply chain.
Control programme for animal feeds. Approval and registration of abattoirs and other meat
Hunting and hunter control. processing establishments, including large and small
Field slaughter depot and farm abattoir approval and meat processors, wholesale and retail facilities.
maintenance. Establishment of procedures and policies for game meat
Small processor identification and approval of the control by the relevant health authorities.
establishments. Establishment of programmes for Good Hygiene
Establishment of programmes for Good Hygiene Practices (GHP’s) as well as Good Manufacturing
Practices (GHP’s) and conducting regular Hygiene Practices (GMPs).
Assessments. Training of slaughter and processing staff.
Implementation of traceability procedures. Training of health authority officials and meat
Water control. examiners.
Training of slaughter and processing staff. Conducting of a Hazard analysis to determine possible
Training of meat examiners. food safety hazards and the control thereof.
Conducting of a Hazard analysis to determine possible Implementation of traceability procedures.
food safety hazards and the control thereof. Programmes for correct packaging and labelling.
the product include (1) the harvesting or hunting methods (shot guns are used frequently)
utilised for bird hunting as well as (2) the trend not to eviscerate the carcasses after hunting
(often with perforated intestines).
Acknowledgements
The authors would like to thank the Confederation of Hunters Associations of South Africa
(CHASA), the South African Hunters and Game Conservation Association (SAHGCA), as
well as Wildlife Ranching South Africa (WRSA) for their support with the distribution of
the questionnaires.
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Game harvesting procedures and their effect on
meat quality: the Africa experience
Department of Animal Sciences, Private bag X1, Matieland, 7602, South Africa; lch@sun.ac.za
Summary
This paper discusses the major methods employed to harvest game on a commercial basis in
South Africa and Namibia. These two countries are presently the major exporters of game
meat from southern Africa. The methods employed are determined by the specific species
and the terrain where these species are found. The wild behaviour and extensive nature of
game species mean that inevitably the mechanics of game meat production are infinitely more
complex than those of domestic production systems where stock can be driven to a central meat
production facility. Unlike with domestic animals, good management practices that minimise
stress during pre-slaughter handling are difficult to employ with wild ungulates since factors
such as terrain, time limitations, weather and the behaviour of specific species will hinder the
efficiency of the harvesting process. As the export and local consumption of game meat in
Africa increases, it is becoming increasingly important to maximise its quality in order for it to
compete with that of domestic species. One of the major quality aspects that can be controlled
through proper management is the use of cropping methods suited to the specific species being
cropped and efficient in minimising ante mortem stress. Relating this ante mortem stress to
the meat quality of wild ungulates is also essential in understanding the importance of the
harvesting process when it comes to the quality of the product being produced.
Keywords: harvesting, meat quality, venison, Africa
1. Introduction
South Africa and Namibia are well-known for their high quality game meat and game meat
products. Tourists often praise this attribute of game meat (Van Schalkwyk and Hoffman,
2010) as it is regularly offered on the menu in restaurants, guest houses and lodges – tourists
wish to consume game meat as part of their African experience (Hoffman et al., 2003). In
both countries, game numbers increased when land owners were granted ownership of the
wild animals found on their property. Namibia’s freehold farmers have had ownership rights
over land and livestock since the early 1900’s, although the commercial rights over wildlife
and indigenous plants had only been given to freehold farmers in 1967. Farmers in communal
areas received the same rights over wildlife much later (1996) when policies were adopted to
promote community-based natural resource management (Barnett and Patterson, 2006). The
implementation of these policies resulted in wildlife being utilised and valued by the private
sector, driving the wildlife sector into a rapid growth phase (Mendelsohn, 2006).

Although South Africa has had no nation-wide census as pertaining to game numbers, it
is estimated that there are more than 5,000 game farms, 4,000 mixed livestock-game farms
which amount to approximately 13% of the total surface area of the country. Springbok
(Antidorcis marsupialis) is the major species harvested and exported (Table 1) from South
Africa (70% of game animals harvested in 2008) and Namibia (80% of game animals harvested
in 2008). In 2000 it was estimated that the gross income generated by game meat sales in
South Africa alone was R20 million (Eloff, 2002). Kudu and blesbok were respectively the
second and third most utilised species for game meat production in South Africa between
2002 and 2004 (Table 1), even though the amount of meat produced from these two species
were far less than that produced from springbok (Patterson and Khosa, 2005). In 2005, it was
estimated that South Africa exported de-boned meat from 160,000 carcasses, predominantly
from springbok (Antidorcus marsupialis), blesbok (Damaliscus pygargus phillipsi) and kudu
(Tragelaphus strepsiceros). Other species such as zebra (Equus burchelli), blue wildebeest
(Connochaetes taurinus), impala (Aepyceros melampus) and gemsbok (Oryx gazella) were
also exported in smaller numbers (Hoffman and Wiklund, 2006). However, due to the global
economic climate, this number had decreased due to a lower international market demand
to slightly over 76,000 in 2009 which was lower than the 85,500 of 2008.
In Namibia there are at least two million head of game (Table 2), a figure roughly similar to
those for cattle, for sheep and for goats (Barnes et al., 2009). Approximately 90% of the wildlife
is located outside formally proclaimed conservation areas. More than 80% of the larger game
species are found on privately owned farms which comprise about 44% of the surface area
of Namibia (Brown, 2008). Currently at least 41% of Namibia is under wildlife management.
Some 60 communal conservancies are now registered bringing the area under communal
conservancy management to about 15.3% of the area of Namibia. State protection comprises
Table 1. Species and numbers of game harvested commercially for meat production in South Africa for
the period 2002 to 2004 (Patterson and Khosa, 2005).
Species 2002 2003 2004
Number Weight (tons) Number Weight (tons) Number Weight (tons)
Springbok 19,252 287,956 25,133 322,030 20,664 307,374

Kudu 733 64,572 256 21,155 646 51,869
Blesbok 811 29,755 31 1,002 1,379 49,241
Black wildebeest 285 22,460 0 0 222 18,744
Zebra 84 14,240 337 64,914 88 16,633
Eland 14 3,282 0 0 82 16,343
Gemsbok 29 2,491 7 820 139 13,537
Impala 117 3,616 28 794 169 4,296
Deer, fallow 51 1,519 1 33 65 1,733
Bushbuck 6 190 1 32 0 0
Blue wildebeest 29 3,005 1 72 0 0
Total 21,457 433,771 25,816 411,080 23,455 479,783

Game harvesting procedures and their effect on meat quality
16.5%, freehold conservancies 6.1%, private protected land 2.1% and community forests and
concessions 1.3% (Brown, 2009).
Table 2. Wildlife numbers in Namibia.
Species Protected Protected Communal Communal Private land Total

areas NVCFa areas SVCFb land NVCFa land SVCFb
Springbok 33,811c 1771 37,150 37,270 621,561 731,563

Kudu 2,063c 1484 1,545 1,000 345,801 351,893
Gemsbok 11,450c 3115 18,670 5,084 350,092 388,411
Red hartebeest 1,468c 115 700 0 122,805 125,088
Eland 1,704c 524 245 0 34,743 37,216
Plains zebra 18,098c 0 20 0 7,303 25,421
Mountain zebra 8,564c 4347 2,130 2,175 55,520 72,736
Ostrich 3,947c 530 2,840 2,020 36,336 45,673
Blue Wildebeest 4,975c 224 470 0 16,623 22,292
Black faced impala 1,500c 0 0 0 1,870 3,370
Common impala 77c 0 385 0 14,980 15,442
Roan 440c 120 95 0 435 1,090
Sable 256c 60 15 0 902 1,233
Lechwe 0 0 250 0 284 534
Tsessebe 0 15 0 0 162 177
Waterbuck 0 0 0 0 4,475 4,475
Buffalo 1,025c 250 90 0 0 1,365
Giraffe 3,683c 229 666 68 5,769 10,415
Warthog 148c 61 40 0 173,866 174,115
Cheetah 706c 149 405 270 2,970 4,500

Leopard 1,970c 430 960 640 4,000 8,000
Lion 574c 23 109 22 0 728
Elephant 9,043c 24 735 155 0 9957

Hippo-potamus 1,262c 0 300 0 0 1,562
Black rhino 816c 43 45 75 134 1,113
White rhino 54c 62 0 0 75 191
Total 107,634 13,576 67,865 48,779 1,800,706 2,038,560
a NVCF = North of the Veterinary Cordon Fence.
b SVCF = South of the Veterinary Cordon Fence.
c Game counts are not representative of the current numbers of wildlife in protected areas.

The major wildlife species in Namibia under consideration for commercial game meat
export are gemsbok, springbok, kudu (Tragelaphus strepsiceros), mountain zebra (Equus
hartmannae), red hartebeest (Alcelaphus buchelaphus) and eland (Taurotragus oryx). The
suitability of these species is not only based on their population numbers, but also on other
factors such as their reproductive performance, the fact that they occur in large herds in easily
accessible regions, their suitability for commercial harvesting and proximity to de-skinning,
de-boning and processing facilities. In Namibia the impact of wildlife use on the economy is
estimated to be some N$ 1.3 billion when the indirect contributions are included as a result
of a multiplier effect of 1.86 (Barnes et al., 2009).
In both South Africa and Namibia tourism is a growing industry. In Namibia tourism is the
strongest driving force behind the growth of the wildlife industry. This sector is envisaged to
grow at 6.9% per annum between 2008 and 2017. Namibia’s Tourism Satellite accounts show
that in 2006 tourism contributed directly and indirectly (through support industries to the
tourism sector) about 71,777 jobs and N$ 6.8 billion to the Gross Domestic Product (GDP).
These tourists expect to see unspoiled habitat, or at least what they perceive to be unspoiled.
At the same time they expect to see an abundance of game animals. However, surplus animals
need to be removed to maintain populations in equilibrium. There are a number of options
available for the game farmer or owner to control or remove surplus game animals such as:
• making use of predators;
• live auctions;
• recreational hunting: trophy hunting and fresh meat (own use, biltong hunting);
• harvesting or harvesting for commercial meat production.
Although predators are high on the non-consumptive tourists’ list to see and photograph, this
form of control is seldom suitable as the fenced game farms are often small and predators then
consume large numbers of game animals which are frequently scarce and expensive animals
(Power, 2002, 2003; Lehman et al., 2008). Until recently live sales were a feasible option for
managing wildlife populations, but auction prices reached a peak (Eloff, 2002) and are about
half of the price obtained for commercial meat sales (Brown, 2008). Trophy hunting earns
more foreign currency for Namibia than it does for South Africa, which makes Namibia one
of the preferred hunting destinations in Africa. Humavindu and Barnes (2003) suggested that
trophy hunting is about five times more important as a contributor to the national economy
in Namibia as South Africa. It is only Tanzania that earns more foreign currency from trophy
hunting than Namibia (Agriforum, 2007).
Harvesting game for commercial meat sales has huge potential (Von la Chevallerie, 1970; Van
Schalkwyk and Hoffman, 2010). Consumers expect the meat products on the market to have
the required nutritional value, be wholesome, fresh, lean and have adequate juiciness, flavour
and tenderness (Dransfield, 2001, 2003; Ngapo and Dransfield, 2006). The export of game
meat from South Africa and Namibia to the European Union is on the increase (Hoffman,
2003; Van Schalkwyk and Hoffman, 2010). Most of the game harvested on large scale is
destined for export. The standards set for processing of game meat are high and enforced
stringently (Hoffman, 2003). Game harvesters hunting game for the commercial meat trade
and meat processing are however food business operators in their own right and responsible

for the safety of the food they deliver to the game meat value chain (Atanassova et al., 2008).
Von La Chevallerie (1970) listed the requirements for successful harvesting of game as follows:
• humanity;
• economy;
• efficiency;
• low wounding percentages;
• low disturbance and scattering;
• selectivity of correct ages and sexes;
• minimal damage to meat;
• ability to bleed carcasses;
• no association with humans.
It is imperative that game animals are handled correctly prior to harvesting and dressing, as
incorrect handling can result in meat that is unwanted (Hoffman, 2001). If a game animal
is killed with minimal stress then there is a normal pH decline and the quality of the meat
is good. However, when the animal is killed after a running period, little glycogen is left in
the muscles and the pH stays high. No lactic acid is formed and the meat is dark, firm and
dry (DFD meat). Meat with a high pH is also prone to bacterial spoilage. If the game animal
runs for a very long period the liver cannot break down the lactic acid fast enough and lactic
acid builds up in the muscle. This results in the denaturation of the muscle protein and the
meat is pale, soft and exudative/watery (PSE meat) (Van Schalkwyk and Hoffman, 2010). The
traditional season for harvesting is usually in winter when animals are normally better fed
and more water is available. Ambient temperatures are cold enough during winter to prevent
carcasses from spoiling before being dressed and cooled. However, if the necessary cooling
facilities are nearby, the season or time of the year does not have to hamper the harvesting
of game animals (Hoffman, 2003).
2. Harvesting techniques
With the onset of fencing and zoning of large areas of land for use as game ranches and
wildlife conservancies, harvesting of wildlife populations has invariably become an integral
component of the management of game ranches and wildlife conservancies. According to
Bothma (1996) there are three broad types of management options for wildlife populations
on a game ranch:
• Conservation: the manipulation of a small or reduced population to increase its density.
• Sustained harvest: the utilisation of a population through sustained yield over a long time.
• Control: the manipulation of a population whose numbers are too high or which has an
unacceptably high growth rate, to stabilise the population or to reduce its numbers.
The type of management option followed will depend on a number of factors, including the
definitive objectives for the use of the ranch as well as the population dynamics of the species
being considered (Bothma, 1996). Sustainable yield of game for meat production could be
accomplished by regular harvesting of game. Maximum sustained yield assumes a certain
constancy of environmental conditions seldom realised in Africa, particularly in arid regions
(Skinner, 1989). For this reason, harvesting programmes should follow the feedback principles

of Stocker and Walters (1984), taking into account vegetation and also ungulate population
numbers as well as age and sex ratios. The harvesting technique applied depends on the
species, its habitat and the vegetation of the area. The harvesting techniques are continuously
being adjusted so as to harvest the most animals in the least amount of time (Mostert, 2007).
Harvesting of ungulates can either be contracted, in which case professional harvesting teams
are brought in, or a farmer may choose to harvest the animals himself. In the former case,
abattoir facilities are customarily provided by the contractor and there is little flexibility in
the scheduling of such activities. If, for example, an opportunity for harvesting is missed, it
may be some time before a contractor is available again (Skinner, 1989). The use of contractors
is also a necessity if the meat is to be exported or sold on the local market, whereas meat for
own use is exempt from such regulations. In this case, the ranch owner is most likely to do
the harvesting himself. For meat to be of export quality, only head and upper neck shots are
acceptable (Hoffman, 2003).
The use of a specific harvesting technique is also important with regard to the stress placed
on the animal prior to death since ante mortem stress has been shown to have deleterious
effects on the meat quality of domestic livestock as well as on that of game (Smith and Dobson,
1990; Veary, 1991; Wiklund et al., 1995; Hoffman, 2000a; Kritzinger et al., 2002; Hoffman
and Wiklund, 2006). Inefficient harvesting techniques also lead to higher labour costs while
inaccurate shooting wastes ammunition and may result in excessively damaged carcasses.
Inefficient harvesting may also lead to lower game productivity as a result of injured and
stressed animals (Ruggiero and Ansley, 1992). The cost of harvesting has been shown to affect
the price of game meat directly (Van Rensburg, 1992). According to Tinley (1972), there are
four general requirements for successful harvesting:
• Instantaneous death: from humanitarian considerations and also because ante mortem
stress causes inferior meat quality.
• Minimum disturbance to the population, e.g. if a number of animals have been harvested
from one herd, it is unwise to continue chasing the same herd as they become excitable
and later unapproachable.
• Throughout the year, daily human activity on foot, frequent passage of motor vehicles and
spotlighting at night without hunting will accustom animals to the rancher’s presence
and activities and ultimately facilitate harvesting.
• If carcasses are required for consumption of fresh meat, they are required to be in an
unspoilt condition, obtainable only with head and upper neck shots.
2.1 Night harvesting
Night harvesting in the field is undoubtedly the most popular method employed in large-scale
game harvesting operations (Veary, 1991; Lewis et al., 1997; Hoffman, 2000a; Kritzinger et
al., 2002; Hoffman an Wiklund, 2006; Le Grange, 2006; Van Schalkwyk and Hoffman, 2010).
As early as 1964, Dassman reported that this method had the greatest success rate (Dassman,
1964). Hunting commences after dark and usually on moonless nights since animals are
difficult to approach in moonlight and because the moonlight reduces the efficiency of the
spotlights used to blind the animals (Le Grange, 2006). Strong spotlights are used to detect and
blind the animals and hunting may continue until dawn the following day to take maximum

advantage of the moonless conditions (Kritzinger et al., 2002). The spotlights immobilise
the animals so that they can be shot from distances ranging from 25 to 100 m. Ruggeiro
and Ansley (1992) found that shots in excess of 150 m, on average, resulted in unacceptable
accuracy. The latter is especially true when light calibre rifles are used and there is a strong
prevailing wind; conditions frequently found out on the South African plains (Karoo) when
plains game species are harvested. During large-scale operations, several marksmen may be
employed at one time, each with a quota of animals to shoot. The marksmen each proceed
in open chase vehicles and often the shooter may also be the driver so as to avoid delayed
reactions from communication errors between drivers and shooters. One or two people
are usually equipped with spotlights. They stand on the back of the vehicles, sweeping the
light over the terrain to detect the animals. Animals are spotted by the reflection from their
retinas and firing should only commence if clear shots are possible (Kritzinger et al., 2002).
Head and upper neck shots are preferred, using high-velocity, small calibre rifles, since these
reduce carcass loss to a minimum (Bothma, 1996; Le Grange, 2006). Von La Chevallerie
and Van Zyl (1971) found that these shots resulted in the least amount of carcass damage in
springbok and impala, while shots in the shoulder and buttocks can contribute up to 20%
and 50% of carcass weight loss, respectively. According to Le Grange (2006), good-quality
telescopic sights are also a necessity, since open sights do not provide the accuracy required,
especially at ranges as far as 100 m.
A study by Lewis et al. (1997) noted that, with a single marksman, the average time that lapsed
between the culling of impala in a herd was 28 seconds, with the maximum time being 3
minutes and 18 seconds and the minimum time being 2 seconds. It must be noted that in this
study there was a high density of animals and the animals had been, as suggested by Tinley
(1972), habituated to humans. Animals need to be collected as soon as possible after being
shot since the darkness may impede the finding of the animals and in areas that harbour large
populations of predators, these animals may compete with the harvesting team in recovering
the carcasses as they become aware of the hunting routine (Le Grange, 2006).
Studies conducted by Hoffman and Ferreira (2000), Kritzinger et al., (2002), Veary (1991) and
Von La Chevallerie and Van Zyl (1971) indicated that the least amount of ante mortem stress
is experienced during night harvesting so that as a result, the use of night harvesting holds
beneficial effects on certain meat quality parameters. Other advantages of night harvesting
include the fact that animals in a herd are less disturbed by the harvesting (Ledger et al.,
1967; Bothma, 1996) and lower night temperatures may allow for better meat quality as well
while it also results in the absence of flies (Bothma, 1996). Disadvantages, on the other hand,
are that wounded animals are more difficult to recover and in the absence of moonlight, the
method is unsuitable in areas of dense bushveld where the vegetation makes it difficult to see
the animals. Night harvesting may also be more expensive than harvesting methods employed
during the day, since labour costs at night tend to be one and a half times those of day costs
(Van Rensburg, 1992). Another disadvantage is that the culling is limited to moonless nights
of two weeks per month and with the growth in the export market, insufficient numbers are
taken off resulting in alternatives such as day shooting having to be implemented (Le Grange,
2006). In addition, night harvesting can prove disadvantageous if shooters and drivers are
unfamiliar with the terrain since navigation is more difficult at night and this in turn may
hinder the predictability of animal movement.

Night harvesting is also limited in its suitability to certain species. In the case of species
that are not sexually dimorphic like the red hartebeest for example, the sexes are difficult
to tell apart (Ruggeiro and Ansley, 1992; Bothma, 1996). Animals such as the kudu are also
not suitable for this method as they are predisposed to look away from the spotlight or to
close their eyes (Joubert, 1983). This is in contrast to springbok and impala, which are ideal
since they have a tendency to remain still once they have been caught in the spotlight (Lewis
et al., 1997; Conroy, 2005). According to Ruggeiro and Ansley (1992), the territorial nature
and relative tameness of gazelles may reduce their flight distance, whereas the more skittish
temperament of wildebeest and zebra, as well as their habit of running in tight groups, makes
them more difficult targets when using this method. These authors also found that the dark
grey colour of wildebeest made finding its head in the telescopic sight more difficult and the
zebra’s stripes made distinguishing one individual from another more difficult, particularly
when moving in a tight group. During night harvesting, harvesting teams also tend to get
tired so that accuracy tends to decline after 23:00 (Ruggeiro and Ansley, 1992) and more
errors are made the longer the harvesting sessions proceed.
2.2 Day harvesting
There are a number of different harvesting methods employed during the day time. A method
commonly applied in commercial harvesting operations is that of hunting from a vehicle in
a method similar as described under night harvesting. Unlike night harvesting, this method
can be used at any time of the month and animals are spotted by two or more people on the
back of the vehicle. A definitive advantage of harvesting during the day is that marksmen
are able to distinguish sexes (even when species are not sexually dimorphic) as well as age
classes and social groupings so that selective harvesting is possible (Bothma, 1996; Hoffman
and Laubscher, 2009a, 2010). This method is easily used with most species of game since
sighting of the animals is less complicated during the daylight hours. Shots can also be fired
at distances in excess of 150 m since animals are more easily distinguishable from each other
and their surroundings than at night time, although it then becomes increasingly important
for marksmen to be well trained in shooting at such distances. These distances also inevitably
increase the chances of prevailing winds blowing the bullet off course and animals being
wounded although during the day, wounded animals as well as carcasses are more easily
located than at night thereby decreasing the risk of shooting losses.
Another method employed during the day although not commonly used, involves the herding
of animals towards shooting lines using scrambler motorcycles, pick-up vehicles or horsemen.
Marksmen position themselves either in camouflaged bunkers or behind bushes where they
are unseen by the approaching animals (Kritzinger et al., 2002). Veary (1991) found that this
method caused the largest amount of stress to the animals and he reported final pH values
for springbok, harvested using this method, that were similar to a value reported by Hoffman
(2000a) for a single severely stressed impala ram. According to Kritzinger et al. (2002), this
method also causes severe sub-dermal abrasions and bruising as a result of the animals
bumping into each other and falling. The effect of the ante mortem stress on the post mortem
pH of the muscle also causes several deleterious meat quality attributes. Field observations
also indicated that this method is also not suited to small, agile species such as warthog, since
they tend to lie down and hide in the thicket when being chased, making it impossible for the

marksmen to spot them. Bothma (1996) observed similar behaviour in bushbuck and nyala,
which tend to run in any direction and try to hide.
2.3 Hide harvesting
This method is employed during the day time and animals are lured to either a drinking hole
or feeding point. Animals are then killed from a nearby hide, using a silenced rifle (Hoffman
and Wiklund, 2006). The method may be used in areas with dense bush where animals
are difficult to locate on foot or by vehicle. It works well with species such as kudu that are
prone to approaching feeding points although it does not work well with other species such
as impala (Hoffman and Wiklund, 2006). This method may be used by ranchers for meat
for own consumption although it is not commonly used in commercial operations since the
off-take rate is very slow.
2.4 Boma harvesting
The technique employed in this method is similar to that of the mass boma capture of animals
for relocation. Game is herded by a helicopter into a large capture boma, where they are then
shot with a light calibre rifle from the ground (Bothma, 1996). Bomas are usually constructed
from dark coloured plastic since animals are reluctant to challenge an apparently solid wall
and because the animals are unable to see through the plastic. This also ensures that as the
animals proceed forward to ‘escape’, they can be moved into separate compartments (Le
Grange, 2006). Wind direction plays a critical role in setting up the boma so that the position
of the front gate must be downwind from the direction from which the animals will approach
(Le Grange, 2006).
Once the animals are herded into the main area of the boma, it is recommended that they
stand for a short period, usually less than 2 hours (Hoffman and Wiklund, 2006). After this,
they are broken up into smaller groups (± 10 animals per group) and moved into smaller
compartments. It is in these smaller compartments that they are shot and this activity usually
takes approximately 60-90 seconds (Hoffman and Wiklund, 2006). According to Le Grange
(2006), it is best if animals are moved into the smaller compartments during the day and
left until night, when shooting commences. This allows them to settle down and accept the
enclosure as well as the intrusion of the light and the marksman later on, and the animals
can then be dispatched off relatively quickly. From there, the animals are removed from the
compartment to a transport truck where they are hung (head hanging down) and after the
vehicle has moved away a short distance, they are exsanguinated. This is done to minimise the
blood spilt in the killing enclosure as this may stress the next batch of animals. The carcasses
are then transported further to a field abattoir set up in the veld (Mostert, 2007).
This method of harvesting has a practical advantage for dense bushveld areas where the
landscape is inaccessible to vehicles. In these areas, the animals can be driven to areas which
are accessible to trucks and refrigeration vehicles (Bothma, 1996). It allows for a large number
of animals to be harvested and processed within a very short time period and ensures that
no wounded animals are left behind. It also allows for a certain level of selectivity in terms
of which animals are harvested thus allowing animals of trophy status or specific breeding

animals or very young animals to be selected and set free. Animals are processed on the spot
and this allows for easy maintenance of hygiene and easier inspection of the carcasses by the
relevant authorities (Le Grange, 2006). The use of light calibre rifles also causes less damage
to the carcass.
No research has been done with regard to the effect of this harvesting method on meat
quality and according to Hoffman and Wiklund (2006), dominant males in a herd may start
fighting with submissive males and may even kill submissive males. This is in agreement with
Bothma (1996) who found that males of certain species, for example impala, kudu, waterbuck,
gemsbok, blue and black wildebeest, red hartebeest and eland, often fight with one another or
with the females soon after being captured and may need to be separated to prevent injury.
Care should also be taken with those species that have horns since panicked animals may
cause serious injuries to those around them. Fighting and pushing between animals may
result in bruising which will negatively affect meat quality.
Capture myopathy may also become a problem if animals were herded over long distances
and for prolonged periods of time. Certain species such as eland bulls, kudu and waterbuck
may cause problems when held in the boma since they are excellent jumpers and may jump
out if they become overly nervous. This is normally overcome by placing of shade net over
the specific boma area. Buffalo do not challenge the plastic at all unless they can see out
or through it (Le Grange, 2006). Certain species are also more amenable to being driven,
for example eland and blesbok, while others, such as the kudu, easily become nervous and
difficult to herd (Ledger et al., 1967; Bothma and Van Rooyen, 2005). Because species such as
impala are naturally large-herd animals, they are also easily herded and captured in a boma
(Furstenburg, 2005). Other methods of harvesting may be preferred over this method since its
set-up may be more costly and labour needs to be trained in an assembly line-type approach
to process the carcasses as quickly as possible (Bothma, 1996; Le Grange, 2006).
2.5 Helicopter harvesting
This method consists of shooting animals from a helicopter during the day time (from an
altitude of around 6 m) using a 12-bore shotgun while a ground team follows in a vehicle
to collect the dead animals (Veary, 1991; Bothma, 1996). Hunting may be carried out with
semi-automatic shotguns so that as many as six animals can be shot in succession as they
run in a line (Le Grange, 2006). Shooting from a helicopter has the advantage of selective
culling as well as being practical in dense bushveld areas where it is difficult to locate animals
on the ground (Rudman, 1983). A quick estimate of the game population can also be made
during the harvesting operation and harvesting can take place over a larger area than would
be possible with boma harvesting (Bothma, 1996).
According to Van Rensburg (1992), this method is the most expensive when compared to
boma harvesting and harvesting from a vehicle and is usually only employed in large scale
commercial operations since it requires high capital investment. This method has been
successfully used for impala, blesbok, springbok and buffalo in Africa as well as red deer in
New Zealand (Le Grange, 2006). Veary (1991) noted similar muscle ultimate pH values as
those obtained during night harvesting, although according to Le Grange (2006), experience

in Zimbabwe has shown that the high levels of adrenaline released in the animals during
shooting and the excessive increase in body temperature result in extremely rapid meat decay
(probably due to high ultimate pH values and the occurrence of dark, firm and dry meat),
usually rendering the carcasses unfit for human consumption.
A high success rate is also dependent on the skill and accuracy of the shooter as well as the
open nature of the terrain so that only head and neck shots can be utilised. According to
Mostert (2007), this may not always be the case and broken legs can sometimes be observed
in species such as springbok that tend to jump when fleeing. Good communication between
the pilot and ground crew is also essential so that carcasses can be recovered quickly. In many
cases, it may be necessary for the ground crew to use hand-help GPS navigational equipment
for locating the carcasses. Carcasses also need to be eviscerated as soon as possible to cool
the carcasses and reduce decay (Le Grange, 2006). Another disadvantage to this method is
that it may inflict unnecessarily high stress (due to exercise) and bruising on the animals as
well as damage to fences when larger animals attempt to escape the property (Rudman, 1983).
Capture myopathy may also become a problem if animals are chased for extended periods
before being shot. This occurs because of over exertion since wild animals are generally not
equipped to run fast over long distances or for long periods of time. Over exertion results in
increased plasma glucose levels and the oxidative capacity of the mitochondria is exceeded
with the intense simultaneous involvement of the majority of the animal’s muscles. In the
resultant hypoxia, anaerobic metabolism in increased, lactate levels raise dramatically, cell
membrane permeability is increased and various intracellular enzymes are released with
a simultaneous reduction in blood pH, i.e. blood acidification (Veary, 1991). Following a
number of chemical changes in the blood, reactions take place in the damaged tissues and
the kidneys become affected as well. The resulting physiological and chemical effects interfere
with the normal functioning of several vital organs. Animals then die either as a result of
kidney degeneration or heart failure (Bothma, 1996). The high acidification in the muscles
of such animals will hold detrimental effects for the meat quality since a high ultimate pH in
meat results in meat with a high spoilage potential and a resulting short shelf-life (Newton
and Gill, 1981).
2.6 Conventional hunting
This method is suited to all game species and can be applied either on foot, from a hide or from
a vehicle. Harvesting should be done during the day time, preferably in the early morning or
late afternoon if there are no cooling facilities available (Tinley, 1972). It is advantageous as it
causes little disturbance, specifically if done on foot which prevents the animals associating
vehicles with hunting, and game do not run much and consequently yield a higher quality of
game meat. Selective harvesting can also take place with regard to age, sex and social grouping
(Bothma, 2006; Hoffman and Laubscher, 2009b). Although it is unethical to hunt game at
waterholes, it is effective for game which occurs in small groups, for example warthogs, for
timid game such as bushbuck and nyala and in dense bushveld areas where walking is virtually
impossible (Bothma, 2006).

2.7 Harvesting losses
According to Von La Chevallerie and Van Zyl (1971), harvesting losses are usually the result
of three factors:
• Loss of meat unfit for human consumption because of bullet damage.
• Animals shot and not recovered.
• A decline in meat quality because of ante mortem stress to which hunted and wounded
animals are subjected.
Wastage of meat due to bullet damage may be prevented by the right placement of shots
(Table 3) as well as by ensuring good marksmanship of the shooter beforehand since the more
difficult shots such as head and upper neck shots are also the most preferred shots (Bothma,
1996). Studies by Hoffman (2000a,b) and Hoffman and Ferreira (2000) found that head shots
resulted in no wastage of meat and high neck shots resulted in less than 2% wastage of meat.
In the meat trade, the neck is also classified as a lower value joint (Hoffman, 2001) so that the
damage done with regards to carcass value by shots through this region is almost negligible.
On welfare grounds, head shots are preferred since they usually result in instantaneous death
while neck shots may result in paralysis and may not render the animal immediately insensible
(Lewis et al., 1997). However, Hoffman and Sales (2007) has shown that in animals such as
the warthog (Phacochoerus africanus), placement of the bullet in a specific area in the brain is
important as incorrect placement may cause excessive kicking movements (and thus muscle
activity) that could result in pale soft exudative (PSE) meat developing.
Traditionally, hunters prefer to shoot animals through the shoulder rather than through the
head or upper neck since it gives the shooter a larger and more stationary target area and less
chance of missing. This type of shot is usually placed at the top of the crease at the back of the
foreleg, in the position where large vital organs such as the heart and lungs can be found as
well as major blood vessels and nerves. A bullet in this area will either hit the heart, resulting
in massive haemorrhage or will hit the lungs and large blood vessels, resulting in lung collapse
and a ‘quick’ death (Hoffman, 2001). Although this shot can result in up to 20% carcass
damage (Table 3), it is the type of shot that is most likely to result in death since, even if the
exact target area is not hit, most shots in this shoulder area will either result in death (even if
Table 3. Bullet damage from shots at various localities as a percentage of total carcass weight (adapted
from Von La Chevallerie et al., 1971).
Locality of the bullet wound Percentage of carcass damaged
Neck 3.18
Neck and shoulder 15.66
Shoulders 20.58
Shoulder and ribs 22.22
Ribs 5.47
Back 12.47
Other (e.g. stomach, hind quarters) 15.61

not instantaneous) or at least severe wounding of the animal causing no or poor mobility. In
the latter case, a second shot should be enough to kill the animal. According to Van Rooyen
et al. (1996), gut shots should also be avoided since contamination of the carcass from the
stomach and intestinal contents may occur, which is unacceptable. The behaviour of the
animals may also affect harvesting losses and Lewis et al. (1997) found that males generally
respond more actively to disturbances than females and show an increase in response when
in breeding herds, leading to a higher percentage of animals being wounded.
Rifle calibre is also important and light calibres are preferred where possible since they cause
the least amount of damage (Hoffman, 2001). An impala should, for example, preferably not
be shot from a short distance with a 7 mm Remington Magnum but rather with a 7 × 57 mm
or a .30-06 with a heavy bullet (Bothma, 1996).
3. Harvesting operations for meat export
3.1 Legislation
There is a difference in definition for large and small game between the South African
legislation and the European Union legislation. The South African legislation (Anonymous,
2000) defines gemsbok, kudu, hartebeest and zebra as large game (Category B) and springbok
as small game (Category C). The definition section in the European Union legislation (EC,
2004c, Annex I) defines small wild game as wild game birds and lagomorphs living freely in the
wild (paragraph 1.7). A lagomorph is any plant eating mammal with two pairs of incisors in
the upper jaw specialised for gnawing, i.e. rabbit and hare. Large wild game is defined as wild
land mammals living freely in the wild that do not fall within the definition of small wild game
(paragraph 1.8). All the species suitable for commercial harvesting in southern Africa would
thus be classified as large wild game in the EU legislation. Certification of exports of animal
and animal products in South Africa and Namibia is done by the competent authority of the
exporting country. The legislations pertaining to the export of game meat to any other country
are governed by the requirements of the particular importing country. Official veterinarians
are required to make certain health attestations which cover both animal health and public
health conditions (Kamwi and Magwedere, 2007).
The following legislations and guidelines are applicable to the export of game meat from
Namibia to South Africa (Kamwi and Magwedere, 2007):
• South African requirements:
– Codex Alimentarius: Recommended International Code of Hygiene Practice for Game
(CAC, 1993).
– Meat Safety Act No. 40 of 2000 (Anonymous, 2000) and draft game meat regulations.
– Veterinary Procedural Notices.
• Namibian requirements:
– Animal Diseases and Parasites Act No. 13 of 1956 and its regulations as amended
(Anonymous, 2005a).
– Prevention of Undesirable Residues in Meat Act No. 21 of 1991 as amended
(Anonymous, 1991).

– Animal Protection Act No. 71 of 1962 as amended (Anonymous, 1962).

– Veterinary Circulars (VC).
The following legislations and guidelines are applicable to the harvesting, dressing and
export of game meat from Namibia and South Africa to the European Union (Kamwi and
Magwedere, 2007):
• Council Decision 79/542/EEC as regards certification requirements for imports into the
community of certain live ungulate animals and their fresh meat (EC, 1979).
• Council Directive 92/45/EEC on public health and animal health problems relating to
the killing of wild game and the placing on the market of wild-game meat (EC, 1992).
• Council Directive 93/119/EC on the protection of animals at the time of slaughter or
killing (EC, 1993).
• Council Directive 96/23/EC on measures to monitor certain substances and residues
thereof in live animals and animal products (EC, 1996).
• Council Directive 98/83/EC on the quality of water intended for human consumption
(EC, 1998).
• Council Directive 2002/99/EC laying down the animal health rules governing the
production, processing, distribution and introduction of products of animal origin for
human consumption (EC, 2002b).
• Commission Regulation No. 178/2002 laying down the general principles and requirements
of food law, establishing the European Food Safety Authority and laying down procedures
in matters of food safety (EC, 2002a).
• Commission Decision 2003/73/EC-amending Decision 97/468/EC as regards the inclusion
of Estonia and Namibia establishments in provisional lists of third country establishments
from which Member States authorise imports of wild game meat (EC, 2003a).
• Council Directive 2003/99/EC on the monitoring of zoonosis and zoonotic agents (EC,
2003b).
• Commission Regulation No. 1441/2007 amending parts of Regulation No. 2073/2005 on
microbiological criteria for foodstuffs (EC, 2007).
• Commission Regulation No. 852/2004 on the hygiene of food stuffs (EC, 2004a).
• Commission Regulation No. 853/2004 laying down specific hygiene rules for food of
animal origin (EC, 2004b).
• Commission Regulation No. 854/2004 laying down specific rules for the organisation
of official controls on products of animal origin intended for human consumption (EC,
2004c).
• Commission Regulation No. 2073/2005 on microbiological criteria for foodstuffs (EC,
2005a).
• Commission Regulation No. 2075/2005 laying down specific rules on official controls for
Trichinella in meat (EC, 2005b).
• Commission Decision 2008/752/EC amending Annexes 1 and 11 to Council Decision
79/542/EEC as regards certification requirements for imports into the community of
certain live ungulate animals and their fresh meat (EC, 2008).

3.2 Areas approved for harvesting
Game should not be harvested from areas which are subject to official prohibition of
harvesting, whether the prohibition is for reasons of conservation, animal health, animal
or plant chemical control, or any other reason (CAC, 1993). Animals may only be harvested
from the OIE recognised Foot and Mouth Disease (FMD) free zone without vaccination.
In Namibia this zone extends south of the Veterinary Cordon Fence which extends from
Palmgrave Point in the west to Gam in the east of Namibia (for bovine, ovine, caprine, wild
and farmed game). The meat should be obtained from animals originating in areas which
are free of OIE (Office International des Épizooties/World Animal Health Organisation)
notifiable diseases prior to slaughter (Kamwi and Magwedere, 2007).
3.3 Requirements for harvesters
Harvesters should note any abnormal condition they detect in the live game animal, or
during the evisceration or bleeding of a game carcass, and such abnormal condition should
be reported to an inspector if that game carcass is taken to a game establishment (CAC, 1993).
Harvesters are required to be registered and must meet certain requirements understanding
the normal disposition of the animal (Anonymous, 2000). For meat exports to the European
Union harvesters must be trained in health and hygiene and must have sufficient knowledge
of the pathology of wild game and of the production and handling of wild game and wild
game meat after harvesting to be able to undertake an initial examination of wild game at
the point of harvesting.
Training of harvesters should cover at least the following subjects (EC, 2004c: Section 4,
Chapter 1, par 1-5):
• The normal anatomy, physiology and behaviour of wild game.
• Abnormal behaviour and pathological changes in wild game due to diseases, environmental
contamination or other factors which may affect human health after consumption.
• The hygiene rules and proper techniques for the handling, transportation, evisceration,
etc. of wild game animals after killing.
• Legislation and administrative provisions on the animal and public health and hygiene
conditions governing the placing on the market of wild game.
Results from a study by Atanassova et al. (2008) concluded that freshly shot game has a
very good hygienic status when all requirements are properly carried out. They detected a
connection between the shooting methods of expertly and non-expertly shot animals and
the occurrence of Enterobacteriaceae which can cause meat spoilage.
3.4 Ante mortem inspection
If an animal is under stress when harvested, the quality and shelf-life of the meat will be
negatively affected. The meat of stressed animals may discolour. Blood supply to the muscles
is increased and this can result in poor bleeding and meat that is not tender. Only game which
passed the ante mortem inspection and that are seemingly alert may be shot (Van Schalkwyk
and Hoffman, 2010). Ante mortem inspections must be carried out by the harvester prior to

the harvesting operation (CAC, 1993; Anonymous, 2000). If no abnormalities were observed
amongst the animals during the examination, no abnormal behaviour was found before
harvesting and there was no contamination to the environment, the trained person must
attach a numbered declaration to the shot animals stating the condition of the animals as well
as the date, time and place of the killing (EC, 2004c: Chapter II, par 4(a)). For meat exports
to the European Union, harvesters must comply with the requirements imposed by the
European Union to permit the monitoring of certain residues and substances in accordance
with Council Directive 96/23/EC (EC, 1996).
3.5 Shooting
Game should be shot in the field in a humane manner (CAC, 1993). Shooting must be done
by a competent marksman, ensuring immediate death. Only head shots are allowed for
commercial harvesting. This is essential to limit decay and contamination of the meat (Van
Rooyen et al., 1996). Game killed with thoracic and abdominal shots are subject to secondary
inspection (Anonymous, 2000: Part V, Section 11.(1)(h), par 61). For meat exports to the
European Union, shooting must be executed in accordance with Council Directive 93/119/
EC (EC, 1993).
3.6 Bleeding
Game intended for commercial purposes must be bled without delay (CAC, 1993) and
preferably within 10 minutes of being shot (Van Schalkwyk and Hoffman, 2010). Blood is an
ideal growth medium for bacteria and when not well-bled, a carcass will deteriorate faster
(Van Rooyen et al., 1996). Bleeding is done by means of severing the jugular vein and carotid
artery on either side of the neck (throat slitting) with a clean sterilised knife. In any event,
the bleeding must be carried out before the animal regains consciousness. All animals which
have been stunned (shot) must be bled by incising at least one of the carotid arteries or the
vessels from which they arise (EC, 2004c: Chapter III, par 1). The knife used for bleeding must
be washed and sterilised before each cut. Large numbers of bacteria are found on the skin of
any animal and contaminate knives when cutting through the skin. A system for sterilising
the knives should be available. Ideal would be water at a minimum of 82 °C, but since this
is sometimes not practical; an approved chemical steriliser in an enclosed holder which is
fitted to the harvesting vehicle should be used instead. When large numbers of game are
harvested several knives should be used to prevent cross-contamination between carcasses.
A two-colour knife system is often recommended to ensure the effective sterilisation of the
knife not in use. Workers bleeding the game must wash their hands between each carcass with
bactericidal (food grade approved) soap and potable warm water (42-45 °C) (Van Schalkwyk
and Hoffman, 2010).
The different categories must be bled in the following ways (Anonymous, 2000: Part V, Section
11.(1)(h), par 62):
• Category A (large animals): may be bled in a lying position.
• Category B (medium animals): on a ramp at a minimum of 20°.
• Category C (small animals): may be bled in a lying position.

Wounded animals requiring a second shot must be condemned if a time period of 10 minutes
is exceeded after the first shooting. All suspect animals, including those that have been
wounded, must be identified and clearly marked. Detained carcasses can either be totally
condemned or passed conditionally. Suspect carcasses must be separated or fully covered in
plastic when transported to the export abattoir for examination by the State Veterinarian. On
the Suspect Carcass Form, which must accompany the consignment to the game handling
facility, the following should be indicated by the Game Meat Examiner (Van Schalkwyk and
Hoffman, 2010):
• species, age, gender and weight of game animal;
• part of carcass affected;
• possible cause of detention;
• additional observation made during hunting.
3.7 Evisceration in the field
If the shooting site is far from the field abattoir, the intestines and stomach should be removed
from the carcass. Care must be taken to ensure that the abdominal cavity and the cut surfaces
are not contaminated with rumen content or dust or dirt (Ebedes and Meyer, 1996). This
should be done within fifteen to twenty minutes after the animal has been shot. A knife with
a rounded cutting edge is needed for this purpose. Evisceration is easier when the carcass is
in a hanging position with the head hanging downwards. It is advisable to staple the cut skin
of the abdominal wall together for transporting (Van Rooyen et al., 1996).
3.8 Transport of harvested game to field abattoir
Game carcasses should be transported to a field abattoir within two hours of bleeding. The neck
slit area must not be contaminated when transporting the carcass to the field abattoir/depot
(Anonymous, 2000: Part V, Section 11.(1)(h), par 65). Vehicles used for harvesting Category C
(small game: Namibian and South African category) game or springbok (considered as large
game by the EU) must be (Van Schalkwyk and Hoffman, 2010):
• designed with a corrosion resistant hanging frame to bleed carcasses in a hanging position;
• designed to provide sufficient space (no heaping) between carcasses to allow effective air
flow for cooling;
• corrosion resistant and free from holes and cracks;
• durable, non-toxic, smooth surfaced and impervious;
• resistant to impact;
• easily cleanable;
• free from equipment or loose objects, other than what is required for the harvesting of
game;
• designed in such a manner that the animal’s feet are not touching the ground while in
transit.
Vehicles used for the harvesting of Category B (medium) game must:

• comply with the requirements above;
• have a hoist and a ramp manufactured at a minimum slope of 20° for hanging of game.

3.9 Partial dressing of the game carcasses at the field abattoir
Carcasses must be transferred from the collecting vehicle to a clean slaughter frame at the
field abattoir in such a manner as to avoid contamination. Labels must be provided for the
identification of each carcass and its organs (Ebedes and Meyer, 1996) since maintaining
traceability is essential for export purposes. At the field abattoir the heads and feet may be
removed provided that it can still be correlated with the carcasses when meat inspection is
done. Horns may be removed with part of the cranium and stored separately (Anonymous,
2000: Part V, Section 11.(1)(h), par 64). In specific cases like the zebra, heads and feet are not
cut off at the field abattoir/depot since the skin has more value when the skin of the head and
feet is also preserved.
Partial evisceration, normally restricted to the removal of the intact gastrointestinal tract,
serves to reduce the weight and bulk of the carcass and to speed cooling. Such removal
should be restricted to those parts which will not increase exposure to contamination to
an unacceptable level and which the controlling authority determines are not required for
inspection. A game carcass should not be skinned or dressed beyond the extent required in
the Codex guideline (CAC, 1993). Incision lines for opening the hide or skin must be spear
cuts from the inside to the outside. A clean sterilised knife must be used. Lactating udders
and reproductive organs are regarded as condemned material and must be removed with
the skin on in such a way as to prevent contamination. Contact with outer surfaces and
soiled equipment must be avoided at all times. Carcasses may not be washed and soiled or
contaminated areas must be cut off (Van Schalkwyk and Hoffman, 2010).
The trachea and oesophagus are cut loose from the surrounding muscles, from the lower jaw
to the breastbone and the diaphragm is cut away from the ribs. The trachea, oesophagus, lungs
and heart (red offal) are hung in enclosed bags next to the carcass (Van Rooyen et al., 1996). It
must be kept identifiable with the carcass of origin until inspection (Anonymous, 2000: Part
V, Section 11.(1)(h), par 65). The game meat inspector at the field abattoir must inspect each
carcass and matching viscera, head and feet and any abnormalities must be noted down in
a report to be forwarded to the game meat abattoir. If a game meat inspector is not available
at the field abattoir/depot, the viscera, heads and feet must be transported with the carcasses
to the game abattoir while maintaining identification between the carcasses and the organs.
Lockable fly-proof containers must be available during evisceration for the collection of
condemned material (Anonymous, 2000: Part V, Section 11.(1)(h), par 66).
Continuous cleaning and sanitation should be practiced throughout the evisceration process
at the field abattoir (Clean-as-you-go). Workers must continuously clean and sanitise with
warm water (82-87 °C) or chemical steriliser (1-2 ppm free chlorine) all hooks, knives, tools
and other equipment used during evisceration to prevent contamination. The temperature
and/or chlorine level of the water must be tested throughout the harvesting process. The
chemical data sheets of all detergents and sanitizers, as well as the dilutions and contact times
thereof must be available on site. When the floor surface becomes covered in blood and dirt
it should be swept clean (Van Schalkwyk and Hoffman, 2010).

3.10 Chilling of game carcasses
Partially dressed carcasses and offal must be chilled within 12 hours of culling to a temperature
not exceeding 7 °C, but when the ambient temperature is more than 15 °C, it must be chilled
within four hours of being killed (Anonymous, 2000: Part V, Section 11.(1)(h), par 67). Where
the ambient temperature is sufficiently low to achieve the required temperature, carcasses
should be placed under refrigeration soon after harvesting, either in a game depot, game
establishment or other specifically approved facility (CAC, 1993).
Veterinary maturation of meat destined for the European market is necessary. This is a control
process whereby the Foot and Mouth virus is deactivated. Carcasses must be submitted to
maturation at a temperature above + 2 °C and below + 7° C for at least 24 hours before de-
boning. All carcasses that have gone through the maturation period should have a pH of less
than 6.0. The maturation period starts when the door of the chiller truck is closed after the last
carcass have been placed in the chiller truck. This requirement is described in Commission
Decision 2008/752/EC amending Annex I and II of Council Decision 79/542/EEC as regards
certification requirements for imports into the Community of certain live ungulate animals
and their fresh meat (EC, 2008).
Maturation of the meat is critical regarding quality. Game has a high metabolic rate and
incomplete maturation may occur. Conditions before shooting may increase metabolism
(Fink, 1992; Kappelhof, 1999). Cooling of carcasses is hampered due to aponeuroses which
firmly surround the muscles and the often, thick hairy skin (Altemeier et al., 1998). If the
glycogen reserves are reduced by stress, meat maturation and acidification can be impaired
resulting in game meat which is tough, has a limited shelf life and a higher pH (Hofmann,
1987; Fink, 1992; Deutz et al., 2000).
3.11 Vehicles transporting harvested game
Vehicles used for the transport of partially dressed carcasses must comply with the standards
for a meat transport truck according to the requirements for Food Premises under the South
African Health Act No. 61 of 2003. If partially dressed carcasses and offal need to be held in
a chiller truck for periods exceeding eight hours, the chiller unit must have the potential to
chill such carcasses to a core temperature of less than 7 °C within 24 hours of being loaded.
Carcasses must hang away from the floor in such a way as to ensure optimal air flow between
carcasses and to avoid contact between skin surfaces and exposed meat of the body cavities.
Edible rough and red offal transported in the same load space as the carcasses must be packed
in closable leak proof containers (or bags) (Anonymous, 2000: Part V, Section 11.(1)(h), par
68). These organs must preferably hang together with the carcasses.
Game meat can only be marketed commercially if it was transported to a game handling
establishment as soon as possible after the ante mortem inspection. The viscera must
accompany the body and must be identifiable as belonging to a given animal (EC, 2004c:
Chapter II, par 3). If no trained person is available to carry out examinations on the body,
then the head (except for tusks, antlers and horns) and all the viscera except for the stomach
and the intestines must accompany the body (EC, 2004c: Chapter II, par 4 (c)). The heaping

of carcasses should be avoided while travelling to the game handling establishment (EC,
2004c: Chapter II, par 6).
3.12 Hygiene control
An adequate supply of chlorinated, drinkable water (river or dam water is not acceptable)
must be available at the field abattoir as well as on the vehicle that transport the carcasses.
All washing should be preferably done with hot (42 °C) running water. No cloths may be
used to dry meat, equipment or hands (Ebedes and Meyer, 1996). An approved Hygiene
Management System must be implemented by the management of the game establishment
which includes hygiene controls for harvesting. Control measures must be taken to ensure
that no contamination of meat and edible products occur. All workers must be trained in
correct harvesting techniques including principles of hygiene practices (Anonymous, 2005b:
Schedule 2, par 4.7.2).
3.13 Receiving of game carcasses at the game processing plant
Flaying and final dressing of the partially dressed game meat carcasses may only be done
in a game meat processing plant. Carcasses must be offloaded and removed to the holding
chillers without delay. In the case where the chiller truck is used to hold carcasses before
dressing, the doors must be closed when not loading out (Anonymous, 2000: Part V, Section
11.(1)(h), par 69).
3.14 Dressing and portioning of carcasses
Incision lines to a hide or skin must be made with a spear cut from the inside to the outside
with a clean sterile knife. Separate knives must be used for cutting the skin and the rest of the
carcass (Van Rooyen et al., 1996). Mechanical flying knives cannot be used for this purpose.
Contact of the exposed meat with platforms, walls, floors, outer surfaces of the hide or skin
and any soiled equipment must be avoided.
All organs received separately at the game processing plant must be available and identifiable
for meat inspection. Final washing with water is allowed to remove bone chips and blood from
the carcass. Substances intended to prevent spoilage by inhibiting the activities of insects, or
the development of bacteria or moulds, may not be applied to the meat unless it applies with
the requirements of the Foodstuffs, Cosmetics and Disinfectants Act No. 54 of 1972. Approved
carcasses may be halved and quartered before or after chilling. Any further cutting must be
done in a approved cutting plant (Anonymous, 2000: Part V, Section 11.(1)(h), par 71. 72, 73).
Un-skinned game carcasses may only be skinned and placed on the market if it was handled
separately from other food before skinning and not frozen. The de-skinned carcass must
undergo a final inspection in accordance with Regulation (EC) No. 854/2004 (EC, 2004c:
Chapter II, par 8). Cutting and boning must be organized in such a way as to prevent or
minimise contamination. Meat intended for cutting must be brought into the workrooms
progressively and as needed. During cutting, boning, trimming, slicing, dicing, wrapping
and packaging, the meat must be maintained at a temperature of not more than 7 °C with

an ambient temperature of not more than 12 °C. Where premises are approved for different
species, cross-contamination must be avoided by separating the operations of different species
in either space or time (EC, 2004c: Chapter V, par 2).
3.15 Controls at the game establishment
The official veterinarian at the game processing plant will verify that the seal of the truck off-
loading the partially dressed carcasses is not broken and that the seal number corresponds
with the seal number as indicated on the Game Harvesting Control Document and that the
amount of partially dressed game carcasses and their tag numbers concur with information
provided. The official veterinarian will also note the temperatures. Continuous thermo-
control recording is however recommended from loading of the carcasses to arrival and
unloading at the game meat handling facility. The recording must provide the accurate
actual time and temperature analyses covering all phases of harvesting and transport (Van
Schalkwyk and Hoffman, 2010).
The primary responsibility for food safety rests with the food business operator (EC, 2004a:
Chapter I, Article I, par 1) and it is necessary to ensure food safety throughout the food chain,
starting with primary production (par 2). Food business operators must therefore establish,
implement and maintain hygiene control procedures based on HACCP (Hazard Analysis
and Critical Control Points) principles (EC, 2004a: Article 5, par 1). This is applicable to the
harvesting of game for meat exports to the European Union. Records must be available of
observations, checks, results, laboratory analyses and corrective actions taken (Anonymous,
2000: Part III, Section 11.(1)(e), par 47). Personnel must be trained in hygiene procedures
and personal hygiene and records thereof must be kept (Anonymous, 2000: Part V, Section
11.(1)(f)). In order to comply with EU regulations on the monitoring of specific residue in
meat (EC, 1996), the testing for residue of Lead and Cadmium in kidneys and liver of game
animals harvested for commercial purposes is compulsory.
4. Conclusions
As the export and local consumption of game meat from Africa increases, it is becoming
increasingly important to maximise its quality in order for it to compete with that of domestic
species. One of the major quality aspects that can be controlled through proper management
is the use of harvesting techniques suited to the specific species being harvesting and the
efficiency of minimising ante mortem stress. It has been shown that the terrain as well as
the specific behaviour of the targeted species influences the harvesting technique(s) to be
employed. Although very little scientific proof exists that the techniques used in the industry
maximise the meat quality, the techniques are efficient and work.
The hygienic handling of the carcasses prior to skinning and de-boning is another crucial
factor when it comes to the quality of the product being produced. The commercial harvesting
teams in South Africa and Namibia are subject to stringent legislations, regulations and

monitoring to ensure a quality game meat product to the discerned consumer. Most of these
were derived from the formal red meat industry and may require further refining.
The harvesting and role of inspection (or lack thereof) in the whole bushmeat trade has not
been discussed at all in this section. Bushmeat is the informal (frequently illegal) harvesting
of wildlife (not only limited to mammals but could include primates, reptiles, birds, etc.) and
the informal trading of the ‘fresh’ meat on the market.
An aspect that warrants further research as pertaining to the different methods of harvesting
the various game species is a more intensive quantification of the effect of the boma harvesting
method on the meat quality of the animals. Aspects within this scenario that require analysis
include the lairage duration (time from chasing to the start of the culling) as well as the
aspects that are linked to this (such as duration of chasing by the helicopter into the boma). It
is obvious that these requirements (and recommendations that are derived from the research)
will differ from species to species. The effect of species and environmental conditions on
the time period before removal of the gut and contents also needs elucidation. Personal
observation have also indicated that the cooling regimes in the chiller trucks need to be
investigated further as there are quality issues (colour stability and drip loss) with the meat
further down the supply chain when the protocols and cold chain are not maintained. This
is of particular importance with the larger species (zebra, wildebeest, eland, etc.) when the
surface area to volume ratio is such that rate of chilling is slow.
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Zoonotic diseases and direct marketing of game
meat: aspects of consumer safety in Germany
Federal Institute for Risk Assessment, Thielallee 88-92, 14195 Berlin, Germany;
niels.bandick@bfr.bund.de
Summary
Direct marketing is a traditional way of distributing game meat. If direct marketing of small
quantities takes place within a radius of 100 km with delivery directly to the consumer
or the local trade an official meat inspection is not required. However, a high level of
responsibility lies with the hunter, which he can only take, provided he has received a sound
education to be refreshed continuously. The risks caused by consumption of game meat are
primarily associated with lack of hygiene during processing and with unrecognised zoonotic
diseases which can be transferred to humans consuming the meat. Important zoonotic
risks in Germany include Trichinellosis, Shiga toxin producing Escherichia coli (STEC),
enterohaemorhagic Escherichia coli (EHEC), Hepatitis E and Tularaemia.
Keywords: distribution, sociological aspects, safety strategies, zoonoses
1. Introduction
According to Regulation (EC) No. 853/2004 (EC, 2004), persons who hunt wild game with
a view to placing it on the market for human consumption must have sufficient knowledge
of the pathology of wild game, of the production and handling of wild game and wild game
after hunting, especially as regards anatomy, physiology, (abnormal) behaviour, of wild game,
pathological changes in wild game which may affect human health after consumption, hygiene
rules/legislation and administrative provisions on animal and public health, and hygiene
conditions governing the placing on the market of wild game.
2. Importance of hunting and wild game consumption in the society
Hunting plays an important part in human culture and represents an ancestral strategy
of survival. Hunting, especially in Europe and particularly in Germany can be seen as a
significant part of the historical development, although – at present – hunting is only carried
out by a small part of the population. Hunting itself and related gains – like the hunting
tradition, language, and techniques, its economic importance and benefits, the actual prize
(the trophy), the hide and meat of the hunted animal – have not only dramatically decreased
in significance in the eyes of today´s society but also faces ostracism in wide circles of the
population, due to the changes in the societal and moral values over the past few decades.
The latter is without doubt geographically influenced and therefore varies markedly from

region to region and is likely particularly valid for urban societies, whilst greater acceptance
is observed among the rural population. This is entirely understandable because hunting
originates from- and is widely carried out in rural areas, whose local population is more
acquainted with this practice: customs which are familiar and experienced on a regular basis
are rarely met with aversion.
The consumer still associates certain exclusivity with the eating of game meat, although
nowadays it is available year round, due to import and storage logistics and can be bought at
moderate prices. In comparison to the constantly increasing consumption of meat from farm
animal species, game meat consumption is relatively insignificant. The per capita consumption
lies at 1.3% (0.8 kg/yr) of the total meat consumption in Germany (NVS, 2008). Yet, the game
meat market is economically relevant. The animal consumption in Germany is about 73,000
tons, whilst the domestic supply with meat from large feral game species approximates 35,000
tons, representing a monetary worth of about 150 million Euros. Exact data on marketing
of game meat are not available, so these figures are based on approximations of the Federal
Association of Game Meat (European Egg, Poultry and Game Association (EPEGA)), which
estimates one half to be distributed through commercial game handling establishments, the
other half by hunters directly supplying to consumers (oral communication EPEGA, 2009).
3. Safety strategies on game meat hygiene
Although the European Feed and Food law explicitly prohibits local direct marketing of game
meat in small amounts to consumers or to retail dealers, this issue falls under the jurisdiction
of national law. Since 2006, European food law regarding game has changed. Hunters (in their
capacity as food business operators) are responsible for the safety of their products. National
German legislation also considers aspects of responsibility and traceability. The intention is
to provide a level of food safety as high as possible and consequently mandatory measures
of hygienic game meat production and handling under proper conditions (e.g. boning in
adequate rooms, chilling as soon as possible to +7 °C) are included.
Safety strategies for handling game meat include establishing efficient surveillance, providing
(continuing) education for hunters as regards hygiene and zoonotic pathogens, assuring
traceability of products by means of documentation and manageable markets, and the
provision of well equipped facilities.
As result of these strategies, in 2007 11% of all shot large wild game was inspected by official
veterinarians (Table 1). A total of 114,000 hunters in Germany participated in continuing
education, which has allowed them to obtain the status of a ‘trained person’. Of the
approximately 350,000 persons in Germany holding a hunting licence, only about 33% have
received such training, although one may assume that those individuals who have completed
their hunting training after 1987 have satisfactory knowledge due to improved training quality
from this year onwards.
The direct marketing of game meat is at present being forcefully propagated through
marketing campaigns by the umbrella organisation for German hunters. The key strategy of

Zoonotic diseases and direct marketing of game meat
Table 1. Statistics of meat inspection of large wild game in Germany 2007 (Statistisches Bundesamt,
2007: Fachserie 3, Reihe 4.3).
Species Number of meat inspection Whole carcass unfit for consumption
Red deer 10,772 91

Fallow deer 9,732 88
Roe deer 75,286 619
Wild boar 92,129 1,222
Others 1,144 38
Total 189,036 (11%)a 2,058 (1%)
Testing on Trichinella 282,442 9
Samples taken by hunters 125,468
a Hunting bag 2007 total of 1,664,824 animals.
the campaign is drawing attention to the unpolluted condition of game and the high quality
of game meat. However, by adopting a direct marketing strategy the hunter takes on a very
high responsibility for consumer safety. This responsibility must be taken seriously to be able
to successfully fulfil expectations.
4. Food chain and traceability
Article 18 Regulation (EC) No. 178/2002 (EC, 2002) makes the traceability of food that is
put on the market the obligation of the food business operators. This system is also well
established in the local direct marketing of game meat. The attachment of an identification
mark to each carcass is obligatory, as is further documentation by way of special forms, once
game is brought to market in some states of the German Federation. However, the initiative
to make this a nationwide uniform procedure in all states was recently overruled in a German
Parliament voting, which outcome is explained by the fact that the handling of game meat
takes place within close and local quarters, and the origin of the game is usually known as it
is received directly from the hunter.
5. Pathogens and health risks
Outbreaks associated with game meat consumption or handling are rarely reported (except
Trichinellosis and Tularaemia, see below). Yet, despite an increasing quality of hygiene
standards, human cases of infections in which game meat is incriminated occur. Major
examples of the transfer of zoonotic agents from wild game to humans include Trichinella,
Escherichia coli (STEC, EHEC), Salmonella, Hepatitis E virus and Francisella tularensis.

5.1 Trichinella – situation in Germany
As in other European countries, this muscle parasite is also found in Germany and affects
various wildlife animals, like martens, foxes and wild boars. Due to insufficient heating of
the meat, the larvae can be transferred to humans and cause allergic reactions, which may
lead to severe illness or death.
There have been reports on human Trichinella outbreaks after the consumption of meat of
wild boars. For instance, in the year 1977 in Ebermannstadt (Bavaria), 69 persons fell ill
after having eaten raw sausages made from wild boar´s meat. Due to the steadily growing
populations of martens, foxes and boars, an increase in Trichinella occurrence in wildlife
animals can be anticipated. According to figures from the meat inspection authorities, more
than 3.697 million wild boars were examined for Trichinella between 1991 and 2006 (Table
2). A total of 186 Trichinella-positive animals were recorded, which represents a prevalence of
0.005%. In Germany, 1 to 10 registered human cases of Trichinellosis occur per year (mostly
imported contamination). Occasionally Trichinella outbreaks occur through meat from wild
boars or domestic pigs (last major outbreak in 2006 in Mecklenburg-Vorpommern affecting
16 persons). In contrast, of the 425.64 million domestic pigs slaughtered between 1997 and
2006 only 1 animal (in the year 2003) was tested positive for Trichinella.
This example for Trichinella indicates the importance of thorough meat inspections, as the only
reliable way to detect a rare infestation of the muscle parasite in an early stage and thus prevent
transfer to the consumer. However, due to its rare occurrence, the awareness of the necessity of
game meat inspection for Trichinella is still very low among hunters. A Trichinella infestation
Table 2. Hunting bag and positive findings of Trichinella (wild boar, 1991-2006).
Year Number Tested (%) Positive Prevalence (%)
1991 312,768 215,494 68.90% 6 0.0028

1992 248,898 160,901 64.65% 12 0.0075
1993 339,242 214,426 63.21% 20 0.0093
1994 313,214 201,442 64.31% 26 0.0129
1995 253,788 179,385 70.68% 13 0.0072
1996 362,214 251,656 69.48% 10 0.0040
1997 281,916 215,926 76.59% 14 0.0065
1998 251,431 192,764 76.67% 12 0.0062
1999 418,667 292,460 69.86% 9 0.0031
2000 350,976 265,417 75.62% 8 0.0030
2001 531,887 389,008 73.14% 4 0.0010
2002 512,050 397,425 77.61% 12 0.0030
2003 470,283 370,187 78.72% 10 0.0027
2004 476,042 390,570 82.05% 11 0.0028
2005 476,645 402,996 84.55% 11 0.0027
2006 287,080 272,258 94.80% 8 0.0029

is not macroscopically visible and the relevance of selecting specific sample locations is
difficult to understand for the lay person. This was significantly better understood by hunters,
once they had been given special training, enabling them to take samples themselves instead of
having to take the whole animal to an inspection site. Unfortunately, the number of authorised
inspection offices is constantly decreasing since the legal requirements for accreditation have
been elevated, as it is not considered worthwhile to start a laboratory, only carrying out a small
number of inspections. In addition, the current regulations stipulate that inspections must
be done using the digestion method instead of the traditional compression method (which
occasionally gives false negative results). Consequently, increasing findings of T. pseudospiralis
are now reported, which subspecies remained undetected in using the compression method.
5.2 Shiga toxin producing Escherichia coli (STEC) in game meat
The degree of STEC contamination in meat from wildlife animals is equal to or higher than
that in food from farm animals. The presence of STEC in food is frequently associated with its
processing, e.g. animal slaughter practices and hygiene playing a major role in contamination
of meat with STEC.
It is not clear if STEC is present in higher proportions in game than in domestic animals or
if high contamination of wildlife meat with STEC is caused by improper slaughter practices.
Few reports are available on wild life as carriers of STEC, and some of these animals serve as
a food source for humans. Surveys show that game holds the 2nd rank of STEC contaminated
meat samples in Germany (9.9%). Most STEC positive samples are found in mutton (11.1%),
beef is far behind (5.2%) and pork samples are least STEC contaminated (0.7%).
5.3 Enterohaemorhagic Escherichia coli (EHEC)
Since 2001, when obligatory registration was instituted in Germany, approximately 1,100
EHEC-infections per year have been registered in humans, equalling 1.4 infections per
100,000 inhabitants, with beef being the primary source of infection.
Findings of the National Reference Laboratory for Epidemiology of Zoonoses at the BfR
have presented another scenario: in year 2002, 3% of game samples showed contamination
with EHEC, in 2005 the quota was 14.8%, i.e. during this time period, the percentage of
contaminated samples of wild game meat was significantly higher than that of beef. According
to present findings as well as information from other research organisations, it appears that
wildlife (as a reservoir), and game meat as a source of possible EHEC infections in humans have
been underestimated. Possibly, not adhering to proper hygienic methods during game meat
processing explains these observations, similarly as is the case with contamination with STEC.
5.4 Hepatitis E
In 2009, 106 cases of hepatitis E were registered in Germany, and there has been a yearly
increase in the number of cases recorded. It was originally assumed that the infections
originated from travels to areas where the virus is endemic. However, it was later found
that cases of infection included persons who had not travelled to risk areas. In recent years,

there have been increased reports of hepatitis E being found in pigs and wild boars in
Europe and Japan, in the latter country being transmitted to humans through consumption
of insufficiently heated wild boar meat. A recent study from Wichmann et al. (2008) has
identified the consumption of meat and intestines of wild boars as an increased risk factor
for hepatitis E infection in Germany.
Recent studies also show that the virus has widely spread among the wild boars population in
Germany (Schielke et al., 2009). The virus was found in 22 of a total of 148 examined animals
(14.9%), albeit with significant regional differences. This was also confirmed by findings of
other scientists.
To date it remains unclear if the detected strains can be directly transmitted to humans and
how easily this occurs. One of the viruses detected in wild boars show a very close link to a
virus from a human hepatitis E case in Germany. This would seem to suggest the possibility
of virus transmission between both hosts in this particular case. However, the low number
of human infections – in consideration to the widely spread hepatitis E virus in wild boars –
indicates that additional factors are necessary for the transmission of the virus or that only
high virus concentrations lead to infection of humans.
Studies are currently being carried out in various different research organisations in Germany,
with the aim to gain further insight in the spreading of the virus in the wild boars and domestic
pig populations, which would allow improving risk assessments of virus transmission between
wild boars, domestic pigs and humans.
5.5 Tularaemia
Tularaemia is an infection, predominantly of hares, rabbits and other rodents (beaver, mice,
rats). Infected hares and rabbits show symptoms of a haemorrhagic septicaemia (blood
poisoning). In addition, deer, wild boars and domestic animals (e.g. sheep, cows, pigs, dogs
and cats) are susceptible to this pathogen. In a recent retrospective study, antibodies for
Francisella bacteria were found in the serum of 3.1% of a total of 763 examined wild boars
in Mecklenburg-West Pomerania, proving that the tularaemia pathogen is present in some
regions in Germany. According to various reports there is a relationship between the contact
with hares and tularaemia infection in humans. For example, two persons were infected in
Ortenaukreis (Baden-Württemberg) towards the end of 2001 after having eaten hare meat.
In both patients the pathogen was detected in blood serum and by immune histological tests.
Following a report from Griesheim (Hessen) in November 2005, at least 6 persons of a hunting
group fell ill after helping with the gutting of the hunted hares. After an incubation period
of a few days, they developed high fever and lymph nodes swellings. Examination in the
University Clinic in Heidelberg confirmed tularaemia infection.

6. Conclusions
Direct marketing of game meat gains importance in Germany even though its role in the
transfer of zoonotic diseases cannot be excluded. In the recent past, many hunters have
received continuing education by which both the level of their knowledge has been increased
and their awareness that hygienic handling is important for the safety of game meat (cooling,
etc.) has been consolidated.
The process of accreditation of laboratories for the testing of Trichinella has lead to a decrease
in the number of laboratories for economic reasons. Consequently – also because of the often
large distances to the still existing inspection laboratories – the willingness to subject wild
boars to Trichinella examination has decreased.
A careful hygienic practice in dissecting and preparing game meat must be promoted, the
thorough washing of hands being one of the most important preventive measures. As most
zoonotic pathogens are sensitive to heat, the safe preparation of game meat (i.e. adequate
heating) offers the best protection against infection.
A proper education of hunters to improve their consciousness for good hygiene practice and
their knowledge about zoonotic diseases would appear to be the best assurance for consumers
when game meat is acquired via direct marketing. Although this is the primary responsibility
of the hunters associations, official food surveillance agencies are in a position to support
such activities.
References
EC (European Commission), 2002. Regulation (EC) No. 178/2002, laying down the general principles and
matters of food safety. O.J. L 31/1.
EC (European Commission), 2004. Regulation (EC) No. 853/2004 of the European Parliament and the Council of
29th of April 2004 containing specific rules for the hygiene of foodstuffs. O.J. L 139/55.
NVS (Nationale Verzehrsstudie II), 2008. Die bundesweite Erhebung zur Ernährungssituation von Jugendlichen und
Erwachsenen / The purpose of the National Nutrition Survey II. Max Rubner-Institut, Bundesforschungsinstitut
für Ernährung und Lebensmittel (Hrsg.). Available at: http://www.was-esse-ich.de/.
Schielke, A., Sachs, K., Lierz, M., Appel, B., Jansen, A. and R. Johne, 2009. Detection of hepatitis E virus in wild
boars of rural and urban regions in Germany and whole genome characterization of an endemic strain.
Virology Journal 6, 58.
Statistisches Bundesamt Deutschland, 2007. Schlachttier- und Fleischuntersuchung bei Tieren inländischer
Herkunft –untersuchte Tiere in Fachserie 3, Reihe 4.3.

Wichmann, O., Schimanski, S., Koch, J., Kohler, M., Rothe, C., Plentz, A., Jilg, W. and K. Stark, 2008. Phylogenetic
and Case Control Study on Hepatitis E Virus Infection in Germany. The Journal of Infectious Diseases 198,
1732-1741.
Other contributions

Dog bites in hunted large game: a hygienic and
economical problem for game meat production
João R. Alberto1, João P. Serejo2 and Madalena Vieira-Pinto1
1Departamento de Ciências Veterinárias, Lab. Inspecção Sanitária, CECAV-UTAD,
Universidade de Trás-os-Montes e Alto Douro, Apartado 1013, 5001-801 Vila Real, Portugal;
joao_c_alberto@hotmail.com
2Câmara Municipal de Idanha-a-Nova, Gabinete Médico Veterinário, 6060-163 Idanha-a-
Nova, Portugal
Summary
Main objectives of this study were to monitor dog bite occurrence in game meat and to
evaluate the damage caused. For this purpose, a total of 526 animals were evaluated: 337 red
deer (Cervus elaphus), 142 wild boar (Sus scrofa), 29 fallow deer (Dama dama), and 18 mouflon
(Ovis musimon), in hunting zones located in the county Idanha-a-Nova (lat 39° 55’N: long 7°
14’W). A total of 100 (19.01%) of the analysed animals had suffered from dog bites. Of those,
64 were classified as level 1, 20 as level 2 (i.e. removal of affected tissues necessary) and 16 as
level 3 (i.e. necessitating total rejection of the carcass). Apart from the animal welfare issue this
study emphasises the hygienic, microbiological and economic relevance of this problem in the
game meat production chain. The necessity of improving dog behaviour during drive hunting
so as to avoid meat rejection, promote animal welfare and game meat hygiene and quality.
Keywords: drive hunting, dog bite
1. Introduction
Large game hunting campaigns, part of them related to ecotourism, are an important source
of revenue and development for a considerable number of Portuguese regions. Among the
large game species hunted in Portugal, red deer and wild boar are most relevant. Hunting
activity and game management are regulated by national law (number 201/2005, 24th of
November 2005). According to this document, large game species may be legally hunted using
procedures such as stand, stalking, battue, drive hunting, and by spear. Of these approaches
the most important method to hunt large game is drive hunting, where in a previously
designated location, the hunter waits for the animal, which is driven by dog packs moved
forward by beaters.
Although dogs are indeed important for large game hunting in Portugal, excessive dog bites
are undesirable side-effects, as the resulting wounds have several negative consequences
including:
• Impairment of animal welfare.
• Quantitative losses in game meat; resulting from rejection of the affected and surrounding
areas, as recommended in the Codex Alimentarius (1994).

• Hygiene defects and, generally, increase of microbiological contamination of game meat.

• Increased risk of dog infection by diseases transferred from wild game, such as tuberculosis,
leptospirosis, Aujeszky’s disease and parasitoses (echinococcosis, cysticercosis), some of
which are also pathogenic to humans (Coye, 1992). The affected animal can also contribute
to the dissemination of diseases to other geographical regions, and thus affect other wild
or domestic animal species.
In addition, the microbiological condition of game meat can be compromised by poor

placement of shots, and by poor carcass evisceration and dressing practices in the field, as
indicated by Gill (2007). The long time period between the hunt and the beginning of carcass
preparation, as well as improper carcass storage temperature, may also adversely affect the
level of meat contamination (Vieira-Pinto et al., 2005). Contributing to this is the infliction
of carcass wounds resulting from dog bites.
The main objectives of this study where to monitor dog bite incidence in game meat and to
evaluate the damage level.
2. Materials and methods
The study was conducted in the county Idanha-a-Nova in the central-eastern part of Portugal
(lat 39° 55’N: long 7° 14’W) with 1,412.7 km2 and 10,561 inhabitants. Located in a plateau
region, it has a large border line with Spain, which is crossed by the river Erges in the east
and the river Tagus in the south. Idanha-a-Nova is a typical Portuguese rural area, with
approximately 50% of land dedicated to agriculture (43% dry agriculture, 9% watered
agriculture, and 5% graze land); 30% are forested areas, mainly oaks, and 13% is shrub land
with sparse vegetation. This region is considered to be one of the best hunting areas in Portugal
with numerous game estates, many of which also breed cattle and sheep in an extensive free
ranging system, grazing in pastures alongside wild artiodactyls.
From November 2008 to February 2009, 20 organised hunts rendering a total of 526 animals
were evaluated, i.e. 337 red deer (Cervus elaphus), 142 wild boar (Sus scrofa), 29 fallow deer
(Dama dama), and 18 mouflon (Ovis musimon).
On every hunting day, before sanitary inspection, the hunted animals were visually evaluated
and classified, according to the extent and depth of dog bites, based on the scale presented in
Figure 1. The grey area represents the damaged carcass site.
3. Results
From the 526 animals analysed, 100 (19.01%) were affected by dog bites. Of those, 64 were
classified as level 1, 20 as level 2 and 16 as level 3 (Figure 1). Affected and surrounding areas
of all animals with level 1 and 2 wounds were partially rejected. Level 3 carcasses were judged
completely unfit for human consumption (Figure 2).

Dog bites in hunted large game
Level 1 Level 2 Level 3
Bite level Level 1 Level 2 Level 3 Total

% 64 20 16 100
Figure 1. Classification of carcass damages through dog bites. Grey areas indicate the size of damaged
tissues related to carcass size. Intensity of alterations through dog bites in affected carcasses (n=100).
Figure 2. Bite level 2 (left) and level 3 (right).
4. Discussion and conclusions
This study emphasises the relevance of this problem in game meat production. According to
Coye (1992) and Talan et al. (1996), dog teeth harbour a considerable quantity of bacteria.
These authors observed that the majority of the microflora cultured from human wounds
contained a mixture of aerobes and anaerobes, mainly derived from the oral ﬂora of the biting
animal. In these studies, both Pasteurella spp. and Staphylococcus spp. have been recognised
as potentially important pathogens.
Meyers et al. (2008) conducted a study to identify the bacteria present in dogs wound as related
to various grades of bites and reported that 16% of the cases were aerobes, 1% anaerobes and

67% a mixture of both (including Clostridium). Pasteurella canis and pyogenic streptococci
were common in infected wounds, whilst Bacillus spp., Actinomyces spp. and the oral
streptococci were usually found in contaminated wounds. Two other studies on dog bite
wounds revealed Staphylococcus intermedius as the most common isolated bacteria, followed
by Streptococcus spp. and coliforms (Kelly et al., 1992; Griffin and Holt, 2001).
Thus, one may assume that the same bacteria commonly found in wounds resulting from
dog bites are present in affected game meat carcasses. Additionally, these wounds constitute
a port of entry for the game skin flora. Coye (1992) and Moreno (2003) reported that the
external layer of the skin has a rich microbial flora that can cross the skin when its integrity
is compromised by any kind of mechanical action, including dog bites. When the bite occurs
while the animal is already dead, bacteria will not spread from the local area. Yet, the microbial
profile of the carcass can be compromised when the animal is bitten when blood circulation
is intact, through which bacteria from the oral cavity can spread to the entire organism.
There is concern that the killing process or deficiencies during evisceration and chill storage
can have a detrimental effect on hygienic quality (Gill, 2007). In this context, carcass wounds
resulting from dog bites represent an additional negative factor that may influence the
microbiological profile of game meat. It is important to draw attention to this problem as it
has practical animal welfare relevance and should be considered in hygienic rules to improve
the level of consumer protection. The latter is highlighted in Regulation (EC) No. 853/2004
(EC, 2004) that lays down specific hygienic rules for foodstuffs. As previously argued by El-
Ghareeb et al. (2009), the introduction of ‘Good Manufacturing Practice’ (GMP) represents
an important element in securing safe and wholesome game meat.
In consideration of our results, GMP should also include stricter rules on dog behaviour
during drive hunting, so as to prevent jeopardising animal welfare and the safety of game
meat obtained by drive hunting practices. The authors believe that the result of our study
should prompt hunters to improve dog behaviour during game drives.
References
Codex Alimentarius, 1994. Carne y productos carnicol – Codigo internacional recomendado para la inspeccion
ante-mortem y post-mortem de animales de matanza y para el dictamen ante-mortem y post-mortem sobre
animales de matanza y carnes. CAC/RCP 41-1993. Vol. 10. Segunda Edicion 241 pp.
Coye, M.J., 1992. Guidelines for the treatment, investigation, and control of animal bites. The State of California
Health and Welfare Agency Department of Health Services, USA, 71 pp.
EC (European Commission), 2004. Regulation (EC) 853/2004 of the European Parliament and of the Council of
29 april 2004 laying down specific hygiene rules for on the hygiene of foodstuffs. O.J. L 139/55.
condition and shelf life of meat from hunted game birds. Eur. J. Wildl. Res. 55(4), 317-323.
Gill, C.O., 2007. Microbiological conditions of meats from large game animals and birds. Meat Sci. 77, 149-160.
Griffin, G.M. and Holt, D.E., 2001. Dog-bite wounds: bacteriology and treatment outcome in 37 cases. J. Am. Hosp.
Assoc. 37(5), 453-460.

Dog bites in hunted large game
Kelly, P.J., Mason, P.R., Els, J. and Matthewman, L.A., 1992. Pathogens in dog bite wounds in dogs in Harare,
Zimbabwe. Vet. Rec. 131, 464-466.
Meyers, B., Schoeman, J.P., Goddard, A. and Picard, J., 2008. The bacteriology and antimicrobial susceptibility of
infected and non-infected dog bite wounds: Fifty cases. Vet. Microbiol. 127, 360-368.
Moreno, B., 2003. Higiene e Inpección de Carnes- vol II. Ediciones Díaz de Santos, S. A. Madrid, Spain, 137 pp.
Talan, D.A., Moran, G.J., Abrahamian, F., Winer, M.R., Citron, D.M. and Goldstein, E.J.C. 1996. The bacteriology
of infected cat and dog bite wounds. Acad. Emerg. Med. 3, 536.
Vieira-Pinto, M.M., Martins, C., Santos, C., Perestrelo-Vieira, R. and Perestrelo-Vieira, H., 2005. Inspecção
Higio-Sanitária de Caça Selvagem. Epidemiologia de algumas doenças. Ciência e Vida Publicações, Odivelas,
Portugal, 130 pp.

Verocytotoxigenic Escherichia coli (VTEC) in wild
ruminants in Germany
Institute of Veterinary Food Science, Justus-Liebig-University Gießen, Frankfurter Str. 92,
35392 Gießen, Germany; andrea.c.bartels@vetmed.uni-giessen.de
Summary
This contribution addresses the role that feral game animals, hunting practices and the
marketing and distribution of game meat may play in the transmission of verotoxinogenic E.
coli (VTEC) to domesticated ruminants and to humans. Recent data generated in Germany
indicate that the epidemiological role of game meat in the spread of VTEC is underestimated.
Keywords: faecal contamination, translocation of bacteria in game meat
1. Introduction
Escherichia coli is a gram negative, facultative anaerobic, non-spore-forming bacterium and

a member of the Enterobacteriaceae (Rolle and Mayr, 1993; NCBI Taxonomy, 2009). E. coli
strains belong to the physiological gut flora of humans and nearly all mammals, guinea pigs
and chinchillas being notable exceptions.
Of a total of about 104-109 cfu/g gut content, E. coli represents a maximum of 1% of the eubiotic
gut flora and is therefore characterised as belonging to the ‘accompanying microflora’ (Rolle
and Mayr, 1993). However, some strains of these gut commensals can be facultative pathogenic.
The importance of verocytotoxigenic foodborne infectious agents was first recognised in 1982,
when the consumption of insufficiently heated hamburgers led to serious illnesses and deaths
in the United States (Riley et al., 1983) and subsequently enterohaemorrhagic Escherichia coli
(EHEC) were described as a new group of enteropathogenic bacteria for the first time (Karch et
al., 2005). E. coli strains of the serotype O157:H7 are considered to represent the prototype of
enterohaemorrhagic E. coli, causing haemorrhagic colitis (HC), haemolytic uraemic syndrome
(HUS) and thrombotic-thrombocytopenic purpura (TTP) in humans. The production of
verotoxins (shigatoxins) is seen as a primary virulence-factor of this group of pathogens,
so they are also termed verotoxin-producing (shigatoxin-producing) E. coli (VTEC/STEC).
In contrast to this virulence-associated nomenclature, the term enterohaemorrhagic E. coli
(EHEC) is based on the resulting clinical symptoms. The differentiation between EHEC and
VTEC/STEC derives from the observation that not all E. coli strains, which can produce
verotoxins, lead to illness in humans. Nevertheless, up to the present day, there is controversy
concerning the distinct definitions of EHEC, VTEC and STEC.
Whilst VTEC infections of farm animals (e.g. cattle, sheep, goats) have been in the focus of
scientific interest since the early 1980’s, the possible epidemiological role of game animals,

especially feral ruminants, has only more recently received increased attention. Particularly
the reports of the Federal German Institute of Risk Assessment (BfR) have prompted the
consideration of game as a primary reservoir of verotoxinogenic E coli, and it is now feared
game meats might represent a source of infection for humans and that their role in the transfer
of human colibacillosis has to date been underestimated (Lehmann et al., 2006, BfR, 2007).
2. Occurrence of VTEC in wild ruminants in Germany

Bülte et al. (2002) tested faecal samples of healthy cattle and found up to ca. 70% animals
within a herd to contain VTEC, which findings are corroborated by other studies (e.g. Naylor
et al., 2005). Consequently, one considers VTEC to belong to the normal intestinal flora, as
they generally do not lead to illness in ruminants, with the possible exception of calves which
occasionally develop clinical symptoms (Baljer et al., 1990).
German studies conducted in the past few decades have shown that VTEC strains can also
be isolated from samples of game. Bülte and Wrocklage (1992), observed that fallow deer
were shedding VTEC at a rate of 10%, all of these eae-negative strains. From meat sampled
from roe deer two serovars (incriminated in human infections) were isolated, which indicates
that insufficiently cooked meats from these game animal species represent a potential VTEC
source (Thoms, 1999; Trumpf et al., 2000). In more recent studies by the reference laboratory
for zoonoses at the German Federal Institute of Risk Assessment (conducted in the period
2002-2006), a large number of game meat samples have been investigated. In 2005, 14.8% of
the samples analysed were found to be positive for verotoxin-producing E. coli, i.e. higher
than the prevalence generally found in beef. Also, VTEC’s such as O26:H11, O128:H2 and
O103:H2 were detected, which serovars are known to possibly cause severe illness in humans
(BfR, 2007). In another German study from 2006, 29 out of 56 faecal samples (51.8%) of wild
ruminants were detected positive for VTEC; among the 13 different O-serogroups isolated
were O21, O128 and O146 strains (pathogenic for humans), albeit none of these were eae-
positive (Lehmann et al., 2006). An overview of the detection of VTEC in wild ruminants in
Germany is presented in Table 1.
Table 1 illustrates that game meat species may represent a potential reservoir for the
transmission of E. coli-strains to domestic ruminants and humans in Germany. A horizontal
transmission of the infection in farm- and wild ruminants is entirely conceivable as both
wild and domesticated ruminants use the same pasture and the former animal category
can contribute to a constant presence of the pathogens in the population. In addition,
human infection may result from the consumption of VTEC contaminated foodstuffs, e.g.
insufficiently cooked game meat (Busch et al., 2007) or herbal foodstuffs, which have been in
contact with droppings of wild animals (Akashi et al., 1994; Thoms, 1999).

Verocytotoxigenic Escherichia coli in wild ruminants in Germany
Table 1. Proof of verocytotoxigenic E. coli (VTEC) of wild ruminants in Germany.
Animal species Number of Number of Number of positive samples (%) Reference

examined faecal examined game
samples meat samples VTEC in general E. coli O157
Fallow deer 100 0 10 (10.0) 0 Bülte and

Wrocklage, 1992
Roe deer 0 53 4 (7.5) no information Thoms, 1999
Different wild 0 from 80 to 160 year 2002 (3.0) no information BfR, 2007
animals species per year year 2005 (14.8)
(general term (not specified) year 2006 (10.0)
‘game meat’)
Red-, roe- and 56 0 29 (51.8) 0 Lehmann et al.,
fallow deer 2006
Red- and fallow 20 (cumulative 0 3 (15.0) 0 Lehmann et al.,
deer in deer park faecal samples) 2006
3. Ports of entry of VTEC in game meat
3.1 Contamination pathways related to hunting practices
It is of interest to consider the possible pathways by which the more common VTEC strains
may be transferred. Relevant factors may include the following: type of hunting, the condition
of the animal before the shot, the location of the shot, the behaviour of the animals after the
shot and the subsequent supply of game.
3.1.1 Type of hunting and the condition of the animal before the shot
The hunter plays an important role in assuring the quality of game meat, not only in the
supplying of game according to best practices, but even in the choice of method of hunting
(Krug, 1998, Deutz et al., 2006). Favouring the entry of VTEC are the so-called movement
hunts (e.g. driven hunt and battue, particularly with the employment of dog packs). In
this type of hunting, game animals often escape (Krug, 1998) and are therefore exposed to
increased stress. Physical activities, such as chasing animals, lead to an increase in the level
of endotoxins (Zucker and Krüger, 1998; Seidler et al., 1999), which in turn, are responsible
for increased permeability of the intestinal tract for bacteria. This could lead to a premortal
endogenous contamination of the organism, even with serious harmful bacteria (Zucker
and Krüger, 1998; Seidler et al., 1999). This mechanism has been reported for slaughter pigs,
which are known to be particularly vulnerable in view of their genetic make-up, the currently
applied husbandry systems and transportation stress. Thus, premortal stress situations may
lead to the translocation of microorganism from the colonised body regions, such as the gut
or locally infected regions, into generally sterile organs and muscles (Zucker and Krüger, 1998;
Fehlhaber and Alter, 1999; Seidler et al., 1999). In addition, it has been shown that such takes

place more frequently, the more pronounced the premortal stress, as the serum bactericidal
activity is adversely affected. Consequently, even post mortem antibacterial activities remain
reduced (Fehlhaber and Alter, 1999).
The physiological dormancy of deer-like animals in winter represents an additional stress

factor. This may trigger an accelerated glycogen breakdown in game animals especially
during the winter months, when their metabolism is lower. The reduced energy turnover, the
scarcity of food and the simultaneous release of endogenous reserves, have to be considered
in choosing the type and the date of hunting (Balfanz, 2007). Hence, in late winter premortal
VTEC contamination is more likely to occur and consequently culling programmes for these
wildlife species should be completed by the end of December (Balfanz, 2007).
3.1.2 Location of the shot and the condition of the animal after the shot
Krug (1998) reported that, whereas stalk hunting of a single animal allows 90% effective chest
hits, this figure decreases to 25% in hunting drives, which conforms to more recent findings
of Deutz et al. (2006). According to the latter study the time interval between killing and
evisceration is generally considerably longer in drive hunts, whilst a major proportion (i.e.
one-third of the animals) are killed by ‘soft shots’ (i.e. injuring the gastrointestinal tract).
Such undesirable shots do not only jeopardise carcass hygiene by the spread of endogenous
gut microflora, but as they are not necessarily instantaneously fatal, such shots may result
in bacteriemia, by which potentially pathogenic organisms (such as prevailing VTEC’s) are
distributed to muscle tissues (Hadlok, 1993; Kappelhoff, 1999; Deutz, 2000).
3.2 Cross-contamination during game meat portioning
Shot game is usually eviscerated on the spot, sometimes under adverse light and weather
conditions. Unless the hunter possesses the necessary skills and has a basic knowledge of game
evisceration and dressing hygiene, this often results in significant carcass contamination.
In addition, special care must be taken during processing of roe deer, which have a looser
connective tissue structure, increasing the risk of bacterial penetration in surrounding tissue
during breaking up and portioning of the carcass (Deutz, 2000). Therefore, especially in this
species, a careful treatment must be warranted.
References
Akashi, S., Joh, K., Tsuyi, A., Ito, H., Hoshi, H., Hayakawa, T., Ihara, I., Abe, T., Hatori, M., Mori, M. and Nakamura,
T., 1994. A severe outbreak of hemorrhagic colitis and hemolytic syndrome associated with Escherichia coli
O157:H7 in Japan. Europ. J. Pediatrics 153, 650-655.
Balfanz, F., 2007. Untersuchungen des Einflusses von Stoffwechselaktivität und sozialen Hierarchien auf die
Stressbelastung beim Rothirsch. Informationsblatt in Stiftung Wald und Wild in Mecklenburg Vorpommern
Nr. 1.
Baljer, G., Wieler, L.H., Bauerfeind, R., Ludwig, S.B. and Mayr, A., 1990. Nachweis von Vero- (Shiga-like-)
toxinbildende E. coli-Keime (VTEC) mittels Zellkulturtest bei durchfallkranken Kälbern. Tierärztl. Umschau
2, 71-78.

Verocytotoxigenic Escherichia coli in wild ruminants in Germany
BfR (Bundesinstitut für Risikobewertung), 2007. Wildfleisch als Quelle für EHEC-Infektionen unterschätzt.
Pressemitteilung 16/2007, 21.08.2007.
Bülte, M. and Wrocklage, V., 1992. Die PCR-Technik zur Ermittlung von verotoxinogenen E. coli-Stämmen bei
Schlacht- und Nutztieren. Proc. 33. Arbeitstagung des Arbeitsgebietes Lebensmittelhygiene der Deutschen
Veterinärmedizinischen Gesellschaft (DVG), Eigenverlag der DVG, Gießen, Germany, pp. 486-495.
Bülte, M., Goll, M. and Stahl, A., 2002. Nachweis von non-O157 VTEC und O157-Stämmen in einer Mutterkuhherde
in Weidehaltung. Proc. 43. Arbeitstagung des Arbeitsgebietes Lebensmittelhygiene der Deutschen
Busch, U., Huber, I., Messelhäusser, U., Hörmansdorfer, S. and Sing, A., 2007. Nachweis Shigatoxin-bildender/
Enterohämorrhagischer Escherichia coli (STEC/EHEC) mittels Real-Time PCR. J. Verbr. Lebensm. 2, 144-148.
Deutz, A., 2000. Die 10 Gebote für die Wildbrethygiene. Tagung für die Jägerschaft; BAL Gumpenstein, Germany.
Deutz, A., Völk, F., Pless, P., Fötschl, H. and Wagner, P., 2006. Wildfleischhygienische Aspekte zu Stöberjagden
auf Rot- und Rehwild. Arch. Lebensmittelhyg. 57, 197-202.
Fehlhaber, K. and Alter, T., 1999. Mikrobielle Folgen prämortaler Belastungen bei Schlachtschweinen.
Fleischwirtschaft 6, 86.
Hadlok, R.M, 1993. Zitat nach Kujawski, O.G., 1993. Den Waidwundschuss riecht und schmeckt man. Jäger 2, 60-61.
Kappelhoff, W., 1999. Wildbrethygiene in der jagdlichen Praxis. Amtstierärztl. Dienst Lebensmittelkontr. 6,
272-276.
Karch, H., Tarr, P.I. and Bielaszewska, M., 2005. Enterohaemorrhagic Escherichia coli in human medicine. Int. J.
Med. Microbiol. 295, 405-418.
Krug, W., 1998. Das Töten von Wild. Amtstierärztl. Dienst Lebensmittelkontr. 5, 238-241.
Lehmann, S., Timm, M., Steinrück, H. and Gallien, P., 2006. Nachweis von STEC im Kot von Hochwild und in
Wildfleischproben. Fleischwirtschaft 4, 93-96.
Naylor, S.W., Gally, D.L. and Low, C.J., 2005. Enterohaemorrhagic E. coli in veterinary medicine. Int. J. Med.
Microbiol. 295, 419-441.
NCBI Taxonomy, 2009. Available at: http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/ wwwtax.cgi?id=1491.
Accessed 15 February 2010.
Riley, L.W., Remis, R.S., Helgerson, S.D., McGee, H.B., Wells, J.G., Davis, B.R., Hebert, R. J., Olcott, E.S., Johnson,
L.M., Hargrett, N.T., Blake, P.A. and Cohen, M.L., 1983. Hemorrhagic colitis associated with a rare Escherichia
coli serotype. New Engl. J. Med. 308, 681-685.
Rolle, M. and Mayr, A., 1993. Medizinische Mikrobiologie, Infektions- und Seuchenlehre für Tierärzte, Biologen,
Agrarwissenschaftler und Interessierte aus benachbarten Fachgebieten. 6th edition, Enke-Verlag, Stuttgart,
Germany, pp. 579-593.
Seidler, T., Mauersberger, J., Krüger, M. and Fehlhaber, K., 1999. Neue Erkenntnisse zu Mechanismen der
belastungsbedingten endogenen Kontamination und Konsequenzen für die Keimstreuung im Schlachtkörper
von Schweinen. Proc. 40. Arbeitstagung des Arbeitsgebietes Lebensmittelhygiene der Deutschen
Thoms, B., 1999. Nachweis von verotoxinbildenden Escherichia coli in Rehfleisch. Arch. Lebensmittelhyg. 50, 52-54.
Trumpf, T., Herzberg, A., Hirsch, J., Jentzen, A., Kramer, B., Rohwer, W., Vogelsang, B. and Stengel, G., 2000.
Nachweis von Shigatoxin bildenden E. coli (STEC). Amtstierärztl. Dienst Lebensmittelkontr. 7, 18-21.
Zucker, B.A. and Krüger, M., 1998. Auswirkungen von Transportbelastungen auf den Endotoxingehalt im Blut
von Schlachtschweinen. Berl. Münch. Tierärztl. Wschr. 111, 208-210.

Microbial quality of venison meat at retail in the UK
in relation to production practices and processes
Philip J. Richards1, Siyu Wu1, David B. Tinker2, Mary V. Howell3 and Christine
E.R. Dodd1
1Universityof Nottingham, Sutton Bonington Campus, Leicestershire LE12 5RD, United
Kingdom; Philip.richards@nottingham.ac.uk
2David Tinker & Associates Ltd., 17 Chandos Road, Ampthill, Bedfordshire, MK45 2LD,
United Kingdom
3Food Standards Agency, Aviation House, 125 Kingsway, London WC2B 6NH, United
Kingdom
Summary
Venison is a popular game meat in the UK with steadily increasing sales. Deer can be wild,
kept in parks or farmed and this affects whether deer are shot in the open or slaughtered
indoors. The distance the carcasses need to be transported affects whether evisceration is
outside or inside and the time before the carcasses can be chilled. Our work aims to identify
best hygienic practice from the different methods used to produce venison meat in the UK.
We have visited five major UK producers of venison and recorded the production practices
and processes used. We have also examined the microbiological quality of retail venison
meat from these producers with the aim of relating food hygiene status to the production
practices employed.
Keywords: Approved Game Handling Establishment (AGHE), venison, Salmonella,

enterobacteriaceae
1. Introduction
Sales of venison (deer meat) in the UK are steadily increasing as consumers consider that deer
are free from the concerns that exist over the intensive production and harvesting systems
associated with many other meats (Mintel, 2008). Within the EU, producers that intend
to retail large quantities of venison, either within the EU or for export, must process deer
carcasses in an approved game handling establishment (AGHE) approved by the competent
authority which in the UK is the Food Standards Agency (FSA). The FSA is an independent
government department established to protect the public’s health in relation to food. There
is a large degree of variation in the practices employed in the production of venison prior to
it being despatched from the AGHE and placed on the market. This study aims to identify
hygienic best practice from the many different methods used to produce venison. We have
visited five major UK producers, whose combined output represents a significant proportion
of the venison retailed (via multiple retail outlets) to the consumer, where we observed the
range of production practices and processes employed. In addition, we have also examined the

Philip J. Richards, Siyu Wu, David B. Tinker, Mary V. Howell and Christine E.R. Dodd
microbiological quality of the venison (steak and diced meat) from these producers at retail
and determined any correlation of product hygiene with the production practices employed.
2. Material and methods
2.1 Observational studies
This study made confidential observational studies of five UK AGHEs that processed deer
for venison.
2.2 Microbial analysis
2.2.1 Sample preparation
Samples of venison were obtained from retail sources where attribution to known AGHEs
was possible, or obtained as a retail pack directly from the AGHE. Samples were attributed
as ‘wild’, ‘farmed’ or ‘farmed & park’ according to the known source of deer for each AGHE
as given in Table 1. It was not possible to source product from AGHE C. Samples for AGHE
E were purchased directly through a farm shop on site (Ei) and through a supermarket (Eii).
Products were limited to those types known to be produced solely from UK venison. Retail
meat (diced meat and steaks) from wild (n=25), farmed (n=16) and farmed & park (n=30) deer
were examined. A test portion of twenty five grams of meat was weighed and homogenised
with 1:10 volume buffered peptone water (Oxoid Ltd., Hamps., UK).
Table 1. Production practices of UK approved game handling establishments (AGHE).
AGHE Source Alive Slaughter Condition of Venison origin

carcass on receipt
Shot
Exsanguinated
Sport shoot
Eviscerated
Head/Neck
Stunned/
Farmed
Skin on
Thorax
Culled
Farm
Wild
Park
A ✓ 8 ✓ ✓ ✓ ✓ ✓ ✓
B ✓ 8 ✓ ✓ ✓ ✓
C ✓ 8 ✓ ✓ ✓ ✓
D ✓ ✓ ✓ ✓ ✓
E ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓

2.2.2 Direct microbial culture methods
The levels of Escherichia coli, Enterobacteriaceae and Staphylococcus aureus in addition to the
total aerobic count were determined according to the appropriate ISO protocol (Anonymous,
2009). Ten-fold serial dilutions of samples were made using Maximum Recovery Diluent
(Oxoid Ltd.) and appropriate dilutions were plated in duplicate. One-way ANOVA and
student’s T-test was performed in order to evaluate possible differences between counts
from samples produced by the differing processes. Where no organisms were recovered on
count plates a value of half the minimum level of detection for the method was used to allow
statistical analysis.
2.2.3 Detection of Salmonella spp.
The remaining buffered peptone water was pre-enriched overnight at 37 °C and sub-cultured
onto MSRV semi-solid agar (Oxoid Ltd.) for a further 24 h at 42 °C. Where growth characteristic
of Salmonella was observed, the culture was inoculated onto XLD agar (LabM Ltd., Lancs.,
UK) and incubated for 24 h at 37 °C. Colonies with typical Salmonella morphology were
confirmed as Salmonella by agglutination with Poly-O A-S antiserum.
3. Results
3.1 Current practices of the UK venison industry
Venison is obtained from three distinct sources of deer (Table 1): (1) wild, unmanaged,
free-ranging herds; (2) domesticated, farmed herds that are reared and husbanded under
controlled conditions; (3) park herds that are fenced-in, may be subject to some limited
husbandry practices and are commonly less cautious of man than wild deer and held in a
more controlled environment so shot placement is generally easier.
Wild and park deer are slaughtered in the open by rifle bullet after a stalked hunt. Wild deer
are shot by both sport and professional hunters, whereas park deer are generally only culled
by expert marksmen. Farmed deer are slaughtered in AGHEs specifically approved to use a
method similar to other red meat species (captive bolt stunning and exsanguination). Apart
from deer slaughtered in an AGHE, a common problem is the lack of an integral cold chain
throughout the whole process. For example, park deer processed in AGHE E were shot in the
open and transported to a well equipped game larder where they were eviscerated, had head/
hooves removed and were placed in a chiller (7 °C) <2 h after being shot. Carcasses were then
detained in the chiller for ~72 h prior to being transported (<3 h) under ambient temperatures
to the AGHE for final dressing.
3.2 Hygienic status of venison products
Salmonella was not isolated from any of the samples examined. E. coli was isolated from 100%
of the samples from wild deer (Figure 1; AGHEs A & B) and from 32% of the samples from
farmed/park deer (AGHE E), but was not isolated from any of the samples from farmed deer

Philip J. Richards, Siyu Wu, David B. Tinker, Mary V. Howell and Christine E.R. Dodd
(AGHE D). Levels of E. coli and Enterobacteriaceae isolated from samples of wild deer meat
were significantly higher than those from farmed/park (diced, P<0.001; steak P<0.01 Figure
1) and farmed venison (steak, P<0.0001). Enterobacterial contamination on farmed/park
steaks was significantly higher than on farmed steaks (P<0.0001). There was no significant
difference between the levels of S. aureus and aerobic colony counts recorded in each of the
groups (P>0.05).
4. Conclusions
Slaughter and/or evisceration of deer within a specialised environment (slaughterhouse or
AGHE) permit a level of control on factors that influence food hygiene. In the field the increased
likelihood of sub-optimal slaughter through poor shot placement and/or evisceration in non-
ideal conditions may lead to carcass contamination which is further complicated by increased
time between slaughter and chilling and/or breaks in the cold chain prior to further dressing
(Sumner et al., 1977). Each of these factors may account for the greater levels of E. coli and
enterobacterial contamination found on venison from wild deer (Atanassova et al., 2008;
Paulsen and Winkelmayer, 2004). However, unlike E. coli and Enterobacteriaceae, aerobic
colony counts and the presence of S. aureus are not correlated with faecal or environmental
contamination but more so with the degree of handling carcasses undergo during dressing
procedures. The similar levels of such contamination found on all venison regardless of origin
may therefore be due to the comparable way that all deer carcasses are skinned, trimmed,
boned-out and prepared for retail when processed at an AGHE. Cross-contamination, for
instance by the use of common butchery boards and manual handling, during the preparation
of venison for retail and the growth of cold tolerant spoilage bacteria may obscure any
differences in the aerobic colony count that existed on as-dressed carcasses.
8
7
Bacterial Contamination
6
(log 10 CFU/g)
5
4
3
2
1
0
A Diced A Steak B Diced D Steak Ei Diced Ei Steak Eii Eii Steak
(n=5) (n=10) (n=10) (n=16) (n=8) (n=16) Diced (n=4)
(n=2)
Figure 1. Bacterial contamination (log10 cfu/g) of venison meat. Escherichia coli (black); Enterobacteriaceae
(dark grey); Staphylococcus aureus (light grey); Aerobic Colony Counts (white). Solid line = 1/2 minimum
level of detection of E. coli/Enterobacteriaceae; dashed line = 1/2 minimum level of detection for S. aureus
and total aerobic count. n = sample size.

References
Anonymous, 2009. International Organization for Standardization. Available at: http://www.iso.org. Accessed
20 November 2009.
Atanassova, V., Apelt, J., Reich, F. and Klein, G., 2008. Microbiological Quality of Freshly Shot Game in Germany.
Meat Sci. 78(4), 414-419.
Mintel, 2008. Poultry and Game Meat - UK - July 2008. Mintel International Group, London, UK.
Paulsen, P. and Winkelmayer, R., 2004. Seasonal Variation in the Microbial Contamination of Game Carcasses in
an Austrian Hunting Area. Eur. J. Wildl. Res. 50(3), 157-159.
Sumner, J.L., Perry, I.R. and Reay, C.A., 1977. Microbiology of New Zealand Farmed Venison. J. Sci. Food Agric.
28(12), 1105-1108.

Detection of Alaria spp. mesocercariae in game
meat in Germany
Katharina Riehn (neé Möhl)1, Knut Große2, Ahmad Hamedy1 and Ernst Lücker1
1Instituteof Food Hygiene, Faculty of Veterinary Medicine, University of Leipzig, An den
Tierkliniken 1, 04103 Leipzig Germany; riehn@vetmed.uni-leipzig.de
2Stadt Brandenburg an der Havel VLMÜV, Klosterstraße 14, 14770 Brandenburg an der
Havel, Germany
Summary
A steadily increasing number of incidental findings of Alaria alata mesocercariae in meat

of wild boars during official Trichinella inspection necessitates the development of a specific
detection method for this parasite. A reliable verification of infested paratenic hosts seems to
be one of the key factors for both understanding the biology of Alaria spp. and determining a
sound risk assessment within the meaning of the consumer’s health protection. Consequently,
our studies concentrate on (1) the verification of suitability of the official digestion methods
for Trichinella spp. for Alaria alata mesocercariae detection in wild boars, (2) development,
optimisation and validation of methods, and, (3) the distribution of the parasites within their
paratenic hosts.
Keywords: Alaria alata mesocercariae, Distomum muscularis suis (DMS), detection method,
predilection sites, magnetic stirrer method for pooled sample digestion, Trichinella
1. Introduction
Distomum musculorum suis (DMS, Duncker, 1896; syn. Agamodistomum suis, Stiles, 1898)
is the mesocercarial stage of the trematode Alaria alata, a small fluke (0.5-1.5 mm) usually
found in the small intestine of various carnivores in the western hemisphere. The life cycle of
this parasite includes freshwater snails (e.g. Helisoma and Planorbis spp.) as first intermediate
hosts. Cercariae emerge from the snails, penetrate tadpoles, and develop into mesocercariae.
A wide range of paratenic hosts can acquire infection by ingesting tadpoles or other infected
paratenic hosts (Figure 1). Dogs, cats, foxes, mink, and other carnivores become infected by
feeding on these animals. The young flukes migrate through various organs of the definitive
host, including the diaphragm and lungs, before reaching the small intestine. Although the
flukes are generally considered to be non-pathogenic for the definitive host, large numbers
may cause pulmonary haemorrhages during migration or enteritis when they mature in the
small intestine.
Katharina Riehn is the recipient of the IRFGMH poster award, kindly made available through a grant of Verein
‘Grünes Kreuz’.

Katharina Riehn, Knut Große, Ahmad Hamedy and Ernst Lücker
Final host
Paratenic hosts 1st Intermediate host
2nd Intermediate host
Figure 1. Life cycle of Alaria alata.
General information about biology, prevalence and pathogenicity of this parasite has been
given in a previous review (Möhl et al., 2009).
In mid-20th century scientists took notice of the potential human health risk posed by this
parasite. Experimental infection of a primate demonstrated that DMS can cause severe
damages within a paratenic host closely related to humans. Since 1973 several reports about
human larval alariosis in North America have been published (Shea et al., 1973; Byers and
Kimura, 1974; Fernandez et al., 1976; Freeman et al., 1976; Beaver et al., 1977; McDonald
et al., 1994; Kramer et al., 1996). Nearly all cases of human alariosis could be linked to the
consumption or handling of game meat (paratenic host) and/or frog legs (second intermediate
host). Nevertheless, the risk for humans was generally ignored or at least postulated to be
negligible until this issue re-emerged in Europe: Jakšić et al. (2002) and Große and Wüste
(2004; 2006) published results on repeated incidental findings of DMS in meat of wild boars
during routine Trichinella inspection in certain areas of Croatia and Germany respectively.
Figure 2 shows Alaria spp. mesocercariae isolated from a wild boar.
In view of deficiencies in methodology, lack of data on prevalence, and the human DMS
cases which were reported in the meantime, an increased scientific attention should be paid
to this subject. Consequently, the German Federal Institute of Risk Assessment concluded
in its opinion (BfR, 2007) that meat which contains Alaria alata mesocercariae should be
regarded as unfit for human consumption. A final statement concerning the health risks for
consumers could not be given due to the lack of information about both the prevalence of
DMS and the suitability of Trichinella inspection methods to detect this parasite in wild boar
meat. Therefore an appropriate diagnostic method for detection of DMS should be developed
in order to acquire further information about the occurrence and the parasite´s importance
in Germany (BfR, 2007).

Detection of Alaria spp. mesocercariae in game meat in Germany
b
c
500 μm
Figure 2. Living Alaria spp. mesocercarial stages with intact viscera (a = oral sucker, b = penetration
glands, c = acetabulum) alongside dead and partly dissoluted DMS after digestion with TIM.
Our studies concentrate on the most pressing questions of (1) the development, optimisation
and validation of methods for reliable DMS detection, (2) the distribution of the mesocerariae
within their paratenic hosts, i.e. identification of potential predilection sites, particularly in
wild boars, and (3) their prevalence in sylvatic populations of animals with respect to their
introduction into the human food chain. Here we present our first results on predilection
sites we obtained by use of the reference method for Trichinella detection in meat samples
as stipulated in specified in Annex I, Chapter I No. 3 of Regulation (EC) No. 2075/2005
(EC, 2005; in the following abbreviated as Trichinella identification method ‘TIM’) and a
modification of this method up to the presentation at IRFGMH conference, Brno, 2009.
In our studies on DMS in wild boars we first applied the reference method for Trichinella
detection in meat samples according to Regulation (EC) No. 2075/2005 (EC, 2005). We
digested the sample material from the carcasses of 20 wild boars in a total of 218 digestions.
The carcasses were dissected and the available muscle tissue totally sampled for detection of
DMS according to 12 anatomically defined sampling sites (pillar of the diaphragm, tongue,
masticatory muscles (Mm. masseter, temporalis, pterygoideus lat.), ‘cheek’ (i.e. various
tissues in the caudoventral region of the head containing among others muscle, connective,
adipose, glandular and lymphatic tissue), neck, foreleg, shoulder, intercostal muscle, loin,
back, abdominal muscles (Mm. rectus abdominis, obliquus externus abdominis, transversus
abdominis), hind leg, adipose tissue) and completely digested. All carcasses and sample
materials were stored at +2 °C until dissection and preparation and examined within 24
hours. In addition to the original protocol for the magnetic stirrer method for pooled sample
digestion as specified in Annex I, Chapter I No. 3 of the Regulation (EC) No. 2075/2005 (EC,
2005) we modified the TIM in order to optimise its efficiency to detect DMS. Numerous

authors (e.g. Pearson, 1956; Shoop and Corkum, 1981; Shoop et al., 1990; Kimber and Kollias,
2000) report, that DMS prefers locations rich in adipose tissue. Since adipose tissues is
undigestable in the HCl/pepsin digestion we tested a method in which Pankreatin© and
bile acid in a magnetic stirrer apparatus are used for the digestion of adipose tissue. For a
50 g pool sample 5 g Pankreatin© and 1 g bile acid is added to a 2 litre cylinder containing
1.0 litre of tap water, preheated to 37 °C; a stirring rod is placed in the beaker, the beaker is
placed on the preheated plate, the stirring is started and NaHCO3 was added until pH 8 (±
0.5) is reached. 50 g of sample material is chopped in the blender and afterwards transferred
to the 2 litre cylinder. After 60 min the stirrer is switched off and the digestion fluid is poured
through a sieve (mesh size 180 μm). The digestion fluid is allowed to stand for 30 minutes
and subsequently a 40 ml sample of digestion fluid is quickly run off into a small measuring
cylinder. The 40 ml sample is allowed to stand for 10 minutes. A portion of 30 ml supernatant
is then carefully withdrawn by suction to remove the upper layers and leave a volume of not
more than 10 ml. The remaining 10 ml sample of sediment is poured into a larval counting
basin or petri dish. The cylinder is rinsed with not more than 10 ml of tap water, which has
to be added to the sample in the larval counting basin or petri dish. Afterwards, the sample
is examined by a trichinoscope or stereo-microscope at a 15-20× magnification. To date, 89
single samples have been analysed by this lipid digestion method. In parallel, distribution
patterns of DMS in 35 positive wild boars were analysed.
3. Results and discussion
The carcases of 20 wild boars were examined by use of TIM. In 14 cases (70%) DMS was
demonstrated in one ore more of the analysed tissues. The number of DMS isolated from
the different tissue samples varied considerably, ranging from 1 to 48 larvae in samples from
11.6 to 125 g. The distribution within the carcasses was heterogenic in all cases and a clear
distribution pattern as known in Trichinella spp. could not be determined. Table 1 shows
the number of DMS as retrieved from the tissue samples by TIM. It is noticeable that in
some highly infested animals, tissues like ‘cheek’ and masticatory muscles show high larval
burdens, whereas the same tissues are completely negative in other DMS-positive animals.
Although nearly all tested body tissues were infested by the parasites, our results show in
accordance with literature that DMS seems to prefer muscular tissue which contains high
amounts of adipose-, connective- and/or glandular tissues. Cheek, neck, intercostal muscle
and shoulder were infested in more than 40% of all positive animals. The larval burden
decreases with smaller amounts of adipose and connective tissues in the sample material.
Tongue, masticatory muscles, and back are infested in 38.5 resp. 28.6% of all positive cases,
abdominal muscle, hind leg and loin in only 7.1%. The foreleg was the only tissue in which
no DMS were detected at all. Furthermore, it became evident, that a large percentage of
mesocercariae die during the pepsin/HCl-digestion, and they did not demonstrate the typical
structure/shape of DMS during stereo-microscopical examination (see Figure 2). The use
of Pankreatin© and bile acid digestion revealed 2 positive carcasses (No. 10 and 12) that
had remained unidentified by conventional Trichinella digestion. Moreover, an increased
number of DMS was isolated in some cases, even though a maximum of only 10-15% of the
sample material was actually digested. In one case DMS could be exclusively detected in the

Table 1. Number of DMS as retrieved from muscle tissue of samples from different locations of the carcass
(n=20) by application of magnetic stirrer method for pooled sample digestion acc. to Regulation (EC) No.
2075/2005 (results of analyses carried out until 11.09.2009).
Carcass 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Pos/all1
Tongue 2 0 n.a.2 0 0 6 0 0 0 0 0 0 0 0 0 5 1 1 0 0 15/19

Masticatory muscle 6 2 n.a. 0 0 2 48 0 0 0 0 0 0 0 1 0 0 0 0 0 59/19
Cheek 2 9 0 1 0 12 30 30 0 0 0 0 0 0 0 3 11 2 0 0 120/20
Neck 2 1 5 0 0 11 1 0 0 0 0 0 3 5 0 0 0 0 0 2 30/20
Shoulder 1 11 0 1 0 16 0 0 0 0 0 0 0 28 0 0 0 2 0 0 59/20
Foreleg 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0/20
Intercostal 1 0 2 0 0 7 6 0 0 0 0 0 0 39 0 0 5 0 0 4 64/20
Back 1 9 0 0 0 3 0 0 0 0 0 0 0 3 0 0 0 0 0 0 16/20
Abdominal 0 0 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3/20
Loin 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1/20
Hind leg 0 0 0 0 0 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4/20
Total 15 32 10 2 0 61 85 30 0 0 0 0 3 75 1 9 17 5 0 6 351/218
1 Pos/all = number of DMS/number of tested samples.
2 n.a. = not analysed.
abdominal adipose tissue. Table 2 shows the number of DMS as retrieved from adipose tissue
by lipid digestion of wild boars’ carcasses.
After the first experiments with this new method it turned out, that the parasites showed
increased survival rates and a distinctly higher vitality after lipid digestion compared to TIM.
On the basis of these promising approaches we also applied modifications of the Pankreatin©
and bile acid digestion in order to optimise its digestion efficiency.
Our results show that DMS distributes heterogeneous throughout the whole body of its
paratenic host. Preferences are mainly identifiable for tissue composition; they further
indicate, in accordance with literature, that DMS prefers muscular tissue which contains
high amounts of adipose-, connective tissues and/or glandular tissues. Sampling as stipulated
Table 2. Number of DMS as retrieved from adipose tissue by application of Pankreatin© and bile acid
digestion (n=20).
Carcass 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Pos/all1
Adipose tissue n.a.2 n.a. n.a. n.a. n.a. 21 2 6 n.a. 4 0 12 29 5 1 4 n.a. n.a. n.a. 0 84/11
1 Pos/all = number of DMS/number of tested samples.
2 n.a. = not analysed.

in Annex I, Chapter I, No. 2 and Annex III (a) of Regulation (EC) No. 2075/2005 (EC, 2005)
is therefore non-applicable for detection of DMS. Especially the muscles of the distal foreleg
were not infested with DMS.
The vastly differing infestation mode and resulting differences in the predilection sites of
Trichinella and Alaria spp. also suggests, that the official digestion method for Trichinella spp.
as laid down by regulation (EC) No. 2075/2005 (EC, 2005) is inadequate for DMS detection
as this method was optimised for the detection of Trichinella in muscular tissue free of all fat
and connective tissues. At the same time our studies have shown that a large percentage of
mesocercariae die during the pepsin/HCl-digestion and their motility, as a major diagnostic
feature, gets lost. In addition, the use of a sieve harbours a certain danger that mesocercariae
could be retained in the mesh or that vital parasites attach to it with their oral suckers. The
promising approaches with the Pankreatin© and bile acid digestion indicate that this method,
possibly in combination with TIM might be more applicable than the original digestion
methods for Trichinella spp.
4. Conclusions
Our results show that the official digestion methods for Trichinella spp. as stipulated in
Regulation (EC) No. 2075/2005 (EC, 2005) are inadequate for the detection of DMS due to
differing infestation modes and tissue preferences of both parasites. In conclusion, we can
state that further studies are imperative with respect to the high potential pathogenicity of
DMS and their presence in the German wild boar population. The development of a reliable
method for detection of the mesocercariae in meat seems to be one of the key factors in this
context. Simultaneously the distribution patterns of the mesocerariae within their paratenic
hosts need to be investigated thoroughly with the view to determine potential predilection
sites in wild boars. Finally the parasites’ prevalence in sylvatic populations of animals needs
to be elucidated (update: Riehn et al., 2010).
Acknowledgements
The authors would like to thank Lia Kieker, Heiko Wellner, Lutz Gumpert and Lina Zeitler
DVM for their valuable technical assistance in the present study. Further, we would like to
express our gratitude for helpful scientific input made by Dr. Karsten Nöckler, Tanja Wüste
DVM, and Petra Kabelitz DVM. This work was financially supported by the German Federal
Ministry of Food, Agriculture and Consumer Protection (BMELV) through the Federal Office
for Agriculture and Food (BLE), grant number 2808HS012.

References
Beaver, P.C., Little, M.D., Tucker, C.F. and Reed, R.J., 1977. Mesocercaria in the skin of man in Louisiana. Am. J.
Trop. Med. Hyg. 26(3), 422-426.
BfR (Bundesinstitut für Risikobewertung), 2007. Wildschweinfleisch kann den gefährlichen Duncker´schen
Muskelegel enthalten. Stellungnahme Nr. 027/2007 des BfR vom 1. Juli 2007.
Byers, B. and Kimura S.J., 1974. Uveitis after death of a larva in the vitreous cavity. Am. J. Ophthalmol. 77(1), 63-66.
EC (European Commission), 2005. Regulation (EC) No. 2075/2005 of 5 December 2005 laying down specific rules
Fernandez, B.J., Cooper, J.D., Cullen, J.B., Freeman, R.S., Ritchie, A.C., Scott, A.A. and Stuart, P.E., 1976. Systemic
infection with Alaria americana (Trematoda). Can. Med. Assoc. J. 115, 1111-1114.
Freeman, R.S., Stuart, P.E., Cullen, S.J., Ritchie, A.C., Mildon, A., Fernandes, B.J. and Bonin, R., 1976. Fatal human
infection with mesocercariae of the trematode Alaria americana. Am. J. Trop. Med. Hyg. 25, 803-807.
Große, K. and Wüste, T., 2004. Funde des Duncker’schen Muskelegels bei der Trichinenuntersuchung mittels
Verdauungsverfahrens. DVG 45. Arbeitstagung des Arbeitsgebiets Lebensmittelhygiene, 28.09-01.10.2004,
Garmisch Partenkirchen.
Große, K. and Wüste, T., 2006. Der Duncker’sche Muskelegel. Funde bei der Trichinenuntersuchung mittels
Verdauungsverfahren. Fleischwirtsch. 86(4), 106-108.
Jakšić, S., Sunčica, U. and Vučemilo, M., 2002. Nachweis von Mesozerkarien des Saugwurms Alaria alata im
Wildschweinfleisch. Z. Jagdwiss. 48, 203-207.
Kimber, K.R. and Kollias, G.V. 2000. Infectious and parasitic diseases and contaminant-related problems of North
American river otters (Lontra canadensis): a review. J. Zoo Wildl. Med. 31(4), 452-472.
Kramer, M.H., Eberhard, M.L. and Blankenberg, T.A., 1996. Respiratory symptoms and subcutaneous granuloma
caused by mesocercariae: a case report. Am. J. Trop. Med. Hyg. 55, 447-448.
McDonald, H.R., Kazacos, K.R., Schatz, H. and Johnson, R.N., 1994. Two cases of intraocular infection with Alaria
mesocercaria (Trematoda). Am. J. Ophtalmol. 117, 447-455.
Möhl, K., Große, K., Hamedy, A., Wüste, T., Kabelitz, P. and Lücker E., 2009. Biology of Alaria spp. and human
exposition risk to Alaria mesocercariae – A review. Parasitol. Res. 105(1), 1-15.
Pearson, J.C., 1956. Studies of the life cycles and morphology of the larval stages of Alaria arisaemoides (Augustine
and Uribe, 1927) and Alaria canis (LaRue and Fallis, 1936) (Trematoda: Diplostomatidae). Can. J. Zool. 34,
295-387.
Riehn, K. Hamedy, A., Große, K., Zeitler, L. and Lücker, E., 2010. A novel detection method for Alaria alata
mesocercariae in meat. Parasitology Res. 107(1), 213-220.
Shea, M., Maberley, A.L., Walters, J., Freeman, R.S. and Fallis, A.M., 1973. Intraretinal larval trematode. Trans.
Am. Acad. Ophthalmol. Otolaryngol. 77(6), 784-791.
Shoop, W.L. and Corkum, K.C., 1981. Epidemiology of Alaria marcianae mesocercariae in Louisiana. J. Parasitol.
67(6), 928-931.
Shoop, W.L., Font, W.F. and Malatesta, P.F., 1990. Transmammary transmission of mesocercariae of Alaria
marcianae (Trematoda) in experimentally infected primates. J. Parasitol. 76(6), 869-873.

Hygiene management systems for commercial
game harvesting teams in Namibia
Diana L. van Schalkwyk and Louw C. Hoffman
University of Stellenbosch, Private Bag X1, Matieland, South Africa; lch@sun.ac.za
Summary
There are more than two million game animals in Namibia, with numbers increasing at a
rate of 20-40% per annum. Around 90% of the country’s wildlife is located outside formally
proclaimed conservation areas whilst more than 80% of the larger game species are found
on privately owned farms comprising 44% of the surface area of the country. The Namibian
wildlife sector offers a commercially viable alternative for generating farm income. The quality
of the meat harvested from game meat depends on several factors such as the skill and attitude
of the hunter, the health of the game animal before being shot, the position of the shot and
the hygienic handling after shooting. Quality aspects further rely on the time before cooling
and the transport and treatment of game carcasses as well as the period prior to cooling.
Food business operators must therefore establish, implement and maintain hygiene control
procedures based on HACCP (Hazard Analysis and Critical Control Points) principles (EC,
2004a) before exports of game meat to international countries are approved.
Keywords: microbiological spoilage, HACCP, Africa
1. Introduction
Namibia is well-known for its high quality game meat and game meat products. Tourists
often praise this attribute of Namibian game meat, as it is often offered on the menu in
restaurants, guest houses and lodges. Namibia has a number of regulations that apply to the
sustainable use of game animals which are applicable when the harvesting of game animals
for commercial game meat production is used to remove excess animals (Nature Conservation
Ordinance, 1975). Countries importing game meat, such as South Africa and the European
Union, also lay down specific rules and regulations whereby countries willing to export game
meat, must abide.
Only harvesting teams registered with the Namibian Directorate of Veterinary Services and
the Ministry of Environment and Tourism are allowed to harvest for the commercial export
of game meat. Each of the harvesting teams should have a well documented and implemented
hygiene management system, as required by the importing country, in place, before the meat
harvested will be allowed to be exported by the competent authority, which is the Directorate
of Veterinary Services in Namibia. Game may only be harvested from the Office International
des Épizooties (OIE) – the world organisation for animal health – recognised foot-and-mouth
disease (FMD) free zone without vaccination. The fresh meat should be obtained from areas
free of FMD and Rinderpest (Kamwi, 2007).

Diana L. Van Schalkwyk and Louw C. Hoffman
2. Hygiene management systems
The primary responsibility for food safety rests with the food business operator (EC, 2004a)
and it is necessary to ensure food safety throughout the food chain, starting with primary
production. Food business operators must therefore establish, implement and maintain
hygiene control procedures based on Hazard Analysis and Critical Control Points (HACCP)
principles (EC, 2004a). This is applicable to the harvesting of game for meat exports to the
European Union and other countries such as South Africa (Anonymous, 2000, 2004).
2.1 Standard Operating Procedures (SOP’S)
Hunters must be trained in health and hygiene and must have sufficient knowledge of the
pathology of wild game, and of the production and handling of wild game and wild game
meat after hunting, to undertake an initial examination of wild game on the spot (EC, 2004b).
Ante mortem inspections must be carried out by the hunter prior to hunting (CAC, 1993).
Only head shots are allowed for commercial harvesting. This is essential to limit decay
and contamination of the meat (Van Rooyen et al., 1996). Game killed with thoracic and
abdominal shots are subject to secondary inspection (Anonymous, 2000, 2004). A farmer
may only employ a night harvesting team for commercial purposes (Nature Conservation
Ordinance, 1975).
Game intended for commercial purposes must be bled within 10 minutes of being shot. Blood
is an ideal growth medium for bacteria and when not well-bled, a carcass will deteriorate
faster (Van Rooyen et al., 1996). Carcasses must be transferred from the collecting vehicle to
a clean slaughter frame in such a manner as to avoid contamination. Labels must be provided
for the identification of each carcass and its organs (Ebedes and Meyer, 1996). Animals should
be partially eviscerated within 20-30 minutes of harvesting. Partial evisceration, normally
restricted to removal of the intact gastrointestinal tract, serves to reduce the weight and bulk
of the carcass and to speed cooling. Chilling must begin within a reasonable period of time
after killing, preferably within 12 hours after the harvest. When the ambient temperature
is more than 12 °C, carcasses must be chilled within 4 hours (Anonymous, 2000, 2004).
Veterinary maturation of meat destined for the European market is necessary. This is a
control process whereby the foot-and-mouth disease virus is deactivated. Carcasses must
be submitted to maturation at a temperature above +2 °C and below +7 °C for at least 24 h
before de-boning (EC, 2008).
Game meat can only be marketed commercially if it was transported in a refrigerated truck
to a registered game handling establishment as soon as possible after harvesting. The red offal
must accompany the body and must be identifiable as belonging to a given animal (EC, 2004b).
2.2 Sanitation Standard Operating Procedure (SSOP’s)
Hygiene management systems for game harvesting teams comprise Sanitation Standard
Operating Procedures for pre-, during and post-operational cleaning and sanitation.
Sterilizers used to sanitise knives contain 10 ppm free chlorine derived from a chemical

Hygiene management systems for commercial game harvesting teams in Namibia
sterilizer. Drinking water is adjusted to a free chlorine level of 1-2 ppm. All equipment,
including the trucks used for harvesting, are cleaned and sanitised.
2.3 Good Hygiene Practices (GHP’s)
Employees handling the game carcasses wear outer garments suitable for hunting and in such
a manner that the clothing protects against contamination. These include jackets, aprons,
rubber boots and hair nets. Employees undergo medical check-ups regularly and abide by a
strict hygiene code. At least one team member is trained as a game meat inspector and also
maintains the records of the hygiene management system.
2.4 Critical Control Points (CCP’s)
A hygiene risk assessment is used to determine Critical Control Points for the game harvesting
process. Typical Critical Control Points defined are:
• Checks on the potability of water from farms where the harvesting take place.
• Checks on faecal contamination of the partially dressed carcasses.
• Checks on temperatures of the carcasses after being loaded into the refrigerator trucks.
The treatment of water to an acceptable chlorine level is essential since most of the time
untreated water from the farms is used during the harvesting of game. Faecal contamination
can result in unacceptable pathogenic bacterial growth. Maturation of the meat (between
+2 °C and +7 °C in 24 h) is critical regarding safety (EC, 2008). The detection of metal
fragments from bullets is not considered as a Critical Control Point. It is controlled by the
Standard Operating Procedure where only head shots are accepted for commercial harvesting.
A study undertaken by Haldimann et al. (2002) concluded that frequent consumption of game
meat has no significant effect on blood lead levels.
3. Conclusions
Hygiene management systems assist game harvesters and processors in ensuring that all
harvesting, transporting, dressing and processing procedures are done under hygienic
conditions. Micro-organisms are mostly responsible for causing severe food poisoning in
humans who eat contaminated meat. Game meat however, has an inherent resistance to
contamination by micro-organisms and this gives it a competitive edge over other types of
meat (Ebedes and Meyer, 1996).
References
Anonymous, 2000. Meat Safety Act. No. 40. Republic of South Africa.
Anonymous, 2004. Red Meat Regulations. Republic of South Africa.
CAC (Codex Alimentarius Commission), 1993. Meat and Meat products including soups and broths. CAC/RCP
29-1983, Rev.1, Joint FAO/WHO Food Standards Programme. Vol. 10, 2nd Edition, pp. 224.

Diana L. Van Schalkwyk and Louw C. Hoffman
Ebedes, H. and Meyer, S.G.H, 1996. Venison for export. In: Bothma, J. Du P. (ed.). Game range management. 3rd
Ed. J.L. van Schaik Publishers, Pretoria, South Africa, pp. 392-396.
of 29th April 2004, containing specific rules concerning official controls on products of animal origin. O.J.
L155/206.
EC (European Commission), 2008. Council Directive 2008/752/EC: Commission Decision of 27 June 2008
amending Annexes I and II to Council Decision 79/542/EEC as regards certification requirements for imports
into the Community of certain live ungulate animals and their fresh meat. O.J. L 261.
Haldimann, M., Baumgartner, A. and Zimmerli, B., 2002. Intake of lead from game meat – a risk to consumers’
health? Eur. Food Res. Technology 215, 375-379.
Kamwi, J.A., 2007. Export of Namibian game meat/carcasses for commercial purposes. Letter addressed to Game
Meat Products Task Team. Ministry of Agriculture, Water and Forestry, Windhoek, Namibia.
Nature Conservation Ordinance, 1975. An Ordinance to consolidate ad amend the Laws relating to the conservation
of nature; the establishment of game parks and nature reserves; the control of problem animals and to provide
for matters incidental thereto. No. 4, Publication of the Ministry of Justice, Windhoek, Namibia, 94 pp.
Van Rooyen, I., Ebedes, H. and Du Toit, J.G., 1996. Meat Processing. In: Bothma, J. Du P. (ed.). Game range
management. 3rd Ed. J.L. van Schaik Publishers, Pretoria, South Africa, pp. 375-377.

Salmonella spp. in wild boar (Sus scrofa): a public
and animal health concern
Madalena Vieira-Pinto1, Luísa Morais1, Cristina Caleja1,2, Patrícia Themudo3,
José Aranha4, Carmen Torres5, Gilberto Igrejas2,6, Patrícia Poeta1 and
Conceição Martins1
1Departamento de Ciências Veterinárias, CECAV-UTAD, Universidade de Trás-os-Montes e
Alto Douro, Apartado 1013, 5001-801 Vila Real, Portugal; mmvpinto@utad.pt

2Departamento de Genética e Biotecnologia, Universidade de Trás-os-Montes e Alto Douro,
Apartado 1013, 5001-801 Vila Real, Portugal

3Laboratório Nacional de Investigação Veterinária, Estrada de Benfica, 701, 1549 Lisboa,
Portugal
4Departamento Florestal, Lab SIG, Universidade de Trás-os-Montes e Alto Douro, Apartado
1013, 5001-801 Vila Real, Portugal

5Biochemistry and Molecular Biology Unit, University of La Rioja, Complejo Cientifico
Tecnologico, C/Madre Dios, 51, 26006 Logroño, La Rioja, Spain

6Institute for Biotechnology and Bioengineering, Centre of Genetics and Biotechnology,
University of Trás-os-Montes and Alto Douro, P.O. Box 202, 5001-801 Vila Real, Portugal
Summary
Wild boars constitute a potential reservoir and may spread zoonotic agents, including
Salmonella sp. and thus they represent a source of infection for (wild and domestic) animals
and humans. During the 2006 hunting season, 77 rectal faecal samples from animals shot
by hunters in Northern Portugal were collected and analysed to determine the prevalence
and serovars of Salmonella sp. in wild boars (Sus scrofa). The results showed that 17 (22.1%)
were positive for Salmonella sp. In these positive samples, the most prevalent serovar was
Salmonella Typhimurium, identified in 11 (64.7%) isolates, followed by Salmonella Rissen
in 6 (35.3%). These results confirm the importance of wild boar as a reservoir of pathogenic
serovars of Salmonella and as a potential risk for humans and livestock and emphasise the
importance of intervention procedures for improving surveillance.
Keywords: wild boar, Salmonella, zoonoses, wildlife, food safety
1. Introduction
Salmonella is defined, both at the European and national level, as being the main responsible
pathogen for foodborne diseases, with approximately 170,000 annually notified human
salmonellosis cases within the European Union (EFSA, 2007). Because of its importance as
a zoonotic agent, extensive surveillance programmes exist in all Member States. However, in
spite of sanitary campaigns, and according to recent literature reviews (Bengis, 2002; Gortázar
et al., 2007), the possibility of persistent cycling of infection in wildlife is real and this could
limit the success of domestic animals disease control programmes. In this context Salmonella

Madalena Vieira-Pinto et al.
may serve as a model for studying the role that wildlife plays as zoonotic agent reservoir and
how this could compromise control programmes in use by veterinary authorities.
Previous studies on the occurrence of Salmonella sp. in wildlife highlight the significance of
game animals as carriers contributing to animal and human contamination, e.g. hedgehogs
(Handeland et al., 2002), wild birds (Refsum et al., 2002), gulls (Wahlström et al., 2003), wild
birds and mammals (Millan et al., 2004) and white-tailed deer (Renter et al., 2006). However,
to date, data on the epidemiological distribution of Salmonella sp. in wild boars are very
limited. In particular, bibliographic references for Portugal are lacking.
Wild boar infected with Salmonella sp. may play an important role in the epidemiology of this
zoonotic pathogen. Salmonella sp. shed in their faeces may be ingested by other wild animals
or by domestic livestock animals, either through direct contact or when these resources are
shared by food animals (specially pastured livestock) or through water cross contamination
(Bengis et al., 2002; Vicente et al., 2002; Renter et al., 2006). This wildlife/domestic animal
interface is observed in rural areas in Northern Portugal, where pasturage of domestic
animals and backyard pig production is a tradition.
Human health risks from wild boar infected with Salmonella sp. arise indirectly from
agricultural areas and vegetable products contamination, through direct animal contact,
during the hunting process and carcass manipulation, or directly from ingestion of
contaminated meat or meat products, such as sausage (Renter et al., 2006).
Considering the importance of wild boars as a major game species in Northern Portugal, as
well as their potential role in transmission of Salmonella sp. to domestic and wild animal
populations with the attendant risk for human health, assessing the prevalence and serotypes
of Salmonella sp. in free-ranging wild boars harvested by hunters was the main objective of
this study.
2.1 Study area
The study area is located in Northern Portugal, where rural and hunting areas comprise
numerous rural settlements (poorly fenced) that are used by local people to produce
vegetables for sale in local markets. The domestic animal production system is characterised
by outdoor production of ruminants and backyard pig production. Within this area, hunting
activity (wild birds, rabbits and hares, wild boars) represents an important contribution to
the development of the local economy, not only in terms of hunting fees but also from the
viewpoint of complementary expenses spent by hunters for lodging, meals and purchases.
2.2 Sampling procedure
During the hunting season of 2005/2006 (December 2005 to February 2006) several hunting
associations from Northern Portugal were contacted with the request to indicate the calendar

Salmonella spp. in wild boar
of their hunting activities so as to allow collecting a representative number of wild boar

samples. After the hunting events, animals were transported to a site where the research team
directly sampled rectal faeces from harvested wild boars (aseptically, i.e. using latex gloves).
From a total of 253 wild boars (Sus scrofa), 77 hunted animals were sampled, whereupon
samples were coded and transported under refrigerated conditions to the laboratory. All
tested animals were subsequently marketed for human consumption.
2.3 Cultural microbiology
All samples were analysed by means of standard culture methods, according to annex D
of ISO standard 6579:2002 applied to Salmonella detection in animal faeces. Isolates of
presumptive Salmonella (1 to 2 colonies from each sample) were confirmed by means of
biochemical tests (Oxidase reaction, Triple Sugar Iron Agar (Oxoid® – CM277), Urea broth
(Merck® – 1.08483), L-Lysine decarboxylation medium (Oxoid® – CM308S)) and serological
agglutination with Poly A-I & Vi antiserum (Difco® – 222641). Salmonella isolates were
serotyped from each positive sample according to the Kauffmann-White scheme (Popoff,
2001) in the LNIV – National Reference Laboratory for Salmonella.
This study is the first report of Salmonella sp. identification in wild boars´ faecal samples in
Portugal, and demonstrates the presence of this microorganism in 17 (22.1%) faecal samples
from 77 harvested wild boars. This prevalence highlights the importance of the wild boar as
reservoir and as a faecal shedder of Salmonella sp., and indicates that faeces from apparently
healthy wild boars can be a source of this pathogen and be potentially transferred, to livestock,
other animals and humans.
Vicente et al. (2002) reported a small seroprevalence of Salmonella in Spain, as based on

antibodies against Salmonella serovar B in 4% and Salmonella serovar C in 3% of the cases.
In contrast, Vengust et al. (2006) reported a seroprevalence of 47% of Salmonella sp. in wild
boars in Slovenia. So far, no data on the prevalence of Salmonella sp. in faecal samples of wild
boars have been published.
In our study, only two serovars were identified: Salmonella Typhimurium (64.7%) and
Salmonella Rissen (35.3%). This is partially explained by the small number (one or two) of
Salmonella sp. colonies isolated and identified per sample, limiting identification of other
possible serovars. Nevertheless, there is a clear dominance of Salmonella Typhimurium
(identified in 64.7% of the positive samples), which suggests this is a predominant serovar in
fecal sample of wild boars, as established in several national (Vieira-Pinto et al., 2005) and
international studies (Davies et al., 2000; Giovannacci et al., 2001; Swanenburg et al., 2001;
Botteldoorn et al., 2004; Castagna et al., 2004), and confirmed by the report on trends and
sources of zoonoses, zoonotic agents, antimicrobial resistance and foodborne outbreaks in
the European Union in 2006 (EFSA, 2007).

The similarity of the Salmonella serovar prevalence pattern of pig and wild boar populations
indicates a possible bidirectional circulation of Salmonella sp. through both animal ecosystems.
This is not unlikely to occur in Northern Portuguese rural areas, as physical contact between
wild boars and backyard raising pigs is frequently reported. Yet, at the present time, this
hypothesis cannot be substantiated since the sanitary status of the northern Portuguese pig
population with respect to Salmonella sp. is not known.
According to Vicente et al. (2002), Salmonella Typhimurium is pathogenic for animals,

and could affect the dynamics of European wild boar populations as well as interfere with
the health of other wild animals, such as birds. In addition, this serovar deserves special
attention in view of its virulence in humans (Botteldoorn et al., 2004) and because it is highly
resistant to antibiotics (Fedorka-Cray et al., 1999; Nielsen et al., 1999; Cruchaga et al., 2001;
Botteldoorn et al., 2004). Currently, Salmonella Typhimurium, is the second most prevalent
Salmonella identified in the European Human Salmonellosis cases in 2006 (EFSA, 2007). In
respect to the second most prevalent serovar, Salmonella Rissen, no references were found
on its prevalence in wildlife.
Wild boars may indirectly represent a reservoir and source of infection for humans, by
uncooked contaminated vegetables (lettuce, cress), water (streams, stagnant pools) or
domestic animals, or directly through contact with infected carcasses, by consuming meat
or meat products (Everard et al., 1979; Kruse et al., 2004; Vengust et al., 2006). Salmonella
sp. occurrence in wild boar meat and carcass was also demonstrated in studies by Decastelli
et al. (1995) in Italy, Kanai et al. (1997) in Japan and Wisniewski (2001) in Poland. For
example, Wisniewski (2001) reported the presence of Salmonella spp. in 11 animals (7%), in
carcasses and internal organs of 156 hunted wild boars. Since hunted wild boars are marketed
for human consumption, they are a potential source of infection to man, if they harbour
Salmonella sp. in their edible tissues, or if meat is contaminated by intestinal content during
evisceration or processing of the carcass (Lillehaug et al., 2005). Faecal contamination of
wild boar carcasses can be expected when hunted animals are poorly bled, eviscerated and
skinned under precarious hygienic conditions (Decastelli et al., 1995; Lillehaug et al., 2005). In
Portugal, this scenario is of particular concern, since the majority of the hunted animals are
used for human consumption, in almost all cases after a deficient transportation of the hunted
animals, and technically and hygienically undesirable evisceration and dressing procedures.
4. Conclusions
The results presented in this study show that wild boar can represent a vehicle of pathogenic
serovars of Salmonella to humans and animals, suggesting that more attention should be paid
to game meat hygiene. Our study also indicates that systematic serological and bacteriological
surveillance of wild boar populations should be improved with a view to better understand
and minimise the impact of such diseases on wild and domestic animals as well as humans.

Salmonella spp. in wild boar
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Decastelli, L., Giaccone, V. and Mignone, W., 1995. Bacteriological examination of meat of wild boars shot down
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Kanai, Y., Hayashidani, K., Kancko, M., Ogawa, M., Takahashi, T. and Nakamura, M., 1997. Occurrence of zoonotic
bacteria in retail game meat in Japan with special reference to Erysipelothrix. J. Food Protect. 60, 328-331.
Kruse, H., Kirkemo, A.-M. and Handeland, K., 2004. Wildlife as source of zoonotic infections. Emerg. Infect. Dis.
10(12), 2067-2071.
Lillehaug, A., Bergsjø, B., Schar, J., Bruheim, T., Vikoren, T. and Handeland, K., 2005. Campylobacter spp.,
Salmonella spp., verocytotoxic Escherichia coli, and antibiotic resistance in indicator organisms in wild cervids.
Acta Vet. Scand. 46, 23-32.
Millán, J., Aduriz, G., Moreno, B., Juste, R.A. and Barral, M., 2004. Salmonella isolates from wild birds and
mammals in the Basque Country (Spain). Rev. Sci. Tech. 23(3), 905-911.
Nielsen, B., Mogelmose, V. and Sorensen, L.L., 1999. Tracing back multiresistant Salmonella Typhimurium DT104
from pork at the slaughterhouse to a specific swine herd by strategical use of serology and culture. Proceedings
of the 3rd International Symposium On the Epidemiology and Control of Salmonella in Pork. Washington,
DC, USA, pp. 261-263.

Popoff, M.Y., 2001. Antigenic formulas of the Salmonella serovars. Report of the WHO Collaborating Centre for
reference and Research on Salmonella. 8th Edition. Institute Pasteur, Paris, France.
Refsum, T., Handeland, K., Baggesen, D.L., Holstad, G. and Kapperud, G., 2002. Salmonellae in Avian Wildlife in
Norway from 1969 to 2000. Appl. Environ. Microbiol. 68(11), 5595-5599.
Renter, D.G., Gnad, D.P., Sargeant, J.M. and Hygnstrom, S.E., 2006. Prevalence and Serovars of Salmonella in
feces of free-ranging White-Tailed Deer (Odocoileus virginianus) in Nebraska. J. Wildl. Dis. 42(3), 699-703.
Swanenburg, M., Urlings, H.A.P., Snijders, J.M.A., Keuzenkamp, D.A. and Van Knapen, F., 2001. Salmonella in
slaughter pigs: prevalence, serovars and critical control points during slaughter in two slaughterhouses. Int.
J. Food Microbiol. 70, 243-254.
Vengust, G., Valencak, Z. and Bidovec, A., 2006. A serological survey of selected pathogens in wild boar in Slovenia.
J. Vet. Med. B 53, 24-27.
Vicente, J., Léon-Vizcaíno, L., Gortázar, C., Cubero, M.J., González, M. and Martín-Antace, P., 2002. Antibodies
to selected viral and bacterial pathogens in European Wild Boars from Southcentral Spain. J. Wildl. Dis.
38(3), 619-652.
Vieira-Pinto, M.M., Themudo, P. and Martins, C., 2005. Occurrence of Salmonella in the ileum, ileocolic lymph
nodes, tonsils, mandibular lymph nodes and carcasses of pigs slaughtered for consumption. J. Vet. Med. B.
52, 476-481.
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of Campylobacter species, VTEC 0157 and Salmonella species in Swedish wildlife. Vet. Rec. 153(3), 74-80.
Wisniewski, J., 2001. The incidence of Salmonella spp. in wild boars in Poland. Medycyny Wet. 57(6), 399-401.
Available at: http://www.medwet.lublin.pl/Year%202001/vol01-06/art224-00htm.htm.

Preliminary results indicating game meat is more
resistant to microbiological spoilage
Louw C. Hoffman1 and Leon M.T. Dicks2
1Department of Animal Sciences, Stellenbosch University, Private Bag X1, Matieland, 7602,
South Africa; lch@sun.ac.za
2Department of Microbiology, Stellenbosch University, Private Bag X1, Matieland 7602,
South Africa
Summary
The aim of this study was to investigate whether game meat has an inherent antimicrobial
activity. To test this, game samples from impala, nyala, warthog, wildebeest, ostrich, zebra and
as controls; beef, mutton and pork were challenged with S. aureus and the growth determined
after overnight incubation at 37 °C. Concluded from the results of this study, meat from game
animals repressed growth of S. aureus much stronger than that of domestic animals. Further
studies to determine whether this phenomenon is applicable to other animals are in progress.
Keywords: microbiological spoilage, Staphylococcus aureus, Africa
1. Introduction
Most foodborne diseases are caused by pathogens such as Escherichia coli, Salmonella,
Campylobacter, Clostridium botulinum and Staphylococcus aureus (Abee et al., 1995). Some
of these microbes originate from soil, water, the intestinal tract of humans and animals. S.
aureus is almost always present in the nose, mouth and skin (Gill and Newton, 1978).
The purpose of this study was to determine whether game meat, in comparison to that of
domestic farm animals such as beef, mutton and pork, had an inherent antimicrobial activity.
There are a number of factors that could cause this apparent phenomenon. The first is the
ultimate pH (pHult) of the meat (Gill and Newton, 1981), which is subsequently the result of
the amount of lactic acid produced from glycogen during anaerobic glycolysis (Aidoo and
Haworth, 1995). When glycogen levels of muscles are depleted due to ante mortem stress the
meat has a high pHult and this increases the likelihood of meat spoilage (Gregory, 1996). Meat
with a high ultimate pH is also classified as dark firm and dry (DFD) and has a high water
binding capacity (Scanga et al., 1998). Game meat that has been stressed ante mortem show
signs of DFD and a strong water binding capacity (Hoffman, 2000). The correlation between
meat water content and microbiological spoilage is well-documented (Gill and Newton, 1981).

Louw C. Hoffman and Leon M.T. Dicks
2. Materials and methods
A naturally occurring Gram- and Catalase-positive isolate from pork, positively identified as
S. aureus was used in the experiments. Approximately 1g grounded meat samples from nine
different animals (sourced from a commercial game meat processor) were each inoculated
with 105 cfu/g (200 μl) of the isolate and 400 μl of sterile distilled water. As controls, 1g of
each meat sample (beef, mutton, pork, impala (Aepyceros melampus), nyala (Tragelaphus
angasii), warthog (Phacochoerus africanus), wildebeest (Connochaetes taurinus), ostrich and
zebra (Equus burchelli)) was inoculated with sterile distilled water (600 μl). The samples were
then incubated at 8 °C for 24 h and tested for the proliferation of the isolate. After incubation
every sample was vigorously mixed with 9 ml sterile distilled water and serial dilutions were
made in duplicate. Dilutions were plated out onto Baird Parker agar, a selective medium for the
identification of S. aureus. Cell counts were determined after overnight incubation at 37 °C.
After determining the cfu/g in each sample on the appropriate medium, the data was plotted
(Figure 1) to reveal the proliferation of the pathogen present in the meat.
In this study beef was taken as the comparative standard, although mutton or pork could
also be chosen. This is done to compare the results obtained from game meat with that of
domestic farm animals. The difference in cell numbers of the S. aureus inoculated beef before
and after 24 hours was designated a value of 100. The resistance of the other meat samples
was then compared with that of beef. Of the other two domestic animals the mutton had a
value of 130 and pork 60. Wildebeest had a value (76) between that of beef and pork. Ostrich
displayed a slightly better value (67). Nyala (44) was fairly better to that recorded for pork.
The impala (31), zebra (17) and especially the warthog (1), showed surprisingly high resistance
to the proliferation of this pathogen.
9 A B C D
8
7
cfu/g (1 x 10 )
5
6
5
4
3
2
1
0
n
la
og
st
ch
a
k
ef
br
al
to
pa
ee
Po
Be
tri
th
Ny
Ze
ut
eb
Im
Os
ar
M
ild
W
Figure 1. Meat experiments done with Baird Parker Agar. The bars marked A represent the meat sample
plus H2O at 0 hours. B indicates the meat sample plus H2O plus the isolate (S. aureus) at 0 hours. The
meat sample plus H2O after 24 hours is indicated by C, and D indicate the meat samples plus H 2O plus
the isolate after 24 hours.

Preliminary results indicating game meat is more resistant to microbiological spoilage
As the meat samples did not contain high numbers of S. aureus before inoculation, the values
recorded at time zero were thus considered negligible.
It would also seem from the results that game meat has a stronger ability to naturally preserve
itself than that of domestic farm animals. It was thought that possible ante mortem stress
might be an influencing factor, but MacDougall et al. (1979) could not find any difference
in microbiological spoilage of farmed young red deer exposed to different levels of ante
mortem stress.
It would appear that the reticulo-endothelial system in some species is more effective than
in others, since venison can be hung for a considerable period without any submission
to decreased temperature or other precaution methods (Lawrie, 1985). Gill et al. (1976)
confirmed that the surviving action of the reticulo-endothelial system destroys bacteria
entering the lymphatic system from the intestines up to 24 hours post mortem.
This phenomenon clearly requires further elucidation.
References
Abee, T., Krockel, L. and Hill, C., 1995. Bacteriocins: modes of action and potential in food preservation and control
of food poisoning. Int. J. Fd Microbiol. 28, 169-185.
Aidoo, K.E. and Haworth, R.J.P., 1995. Nutritional and chemical composition of farmed venison. J. Hum. Nut.
Diet. 8, 441-446.
Gill, C.O. and Newton, K.G., 1978. The ecology of bacterial spoilage of fresh meat at chill temperatures. Meat Sci.
2, 207-217.
Gill, C.O. and Newton, K.G., 1981. The microbiology of DFD fresh meats: a review. Meat Sci. 5, 223-232.
Gill, C.O., Penney, N. and Nottingham, P.M., 1976. Effect of delayed evisceration on the microbial quality of meat.
Appl. Environ. Microbiol. 31, 465-468.
Gregory, N.G., 1996. Welfare and hygiene during preslaughter handling. Meat Sci. 43, 35-46.
Hoffman, L.C., 2000. Meat quality attributes of night-cropped Impala (Aepyceros melampus). S. Afr. J. Anim. Sci.
30, 133-137.
Lawrie, R.A., 1985. Lawrie’s Meat Science. Woodhead Publ. Ltd., Cambridge, UK.
MacDougall, D.B., Shaw, B.G., Nute, G.R. and Rhodes, D.N., 1979. Effect of pre-slaughter handling on the quality
and microbiology of venison from farmed young red deer. J. Sci. Food Agric. 30, 1160-1167.
Scanga, J.A., Belk, K.E., Tatum, J.D., Grandin, T. and Smith G.C., 1998. Factors contributing to the incidence of
dark cutting beef. J. Anim. Sci. 76, 2040-2047.

Section 2
Epidemiology
Key contributions
Trichinellosis in wild and domestic pigs and public
health: a Serbian perspective
Sava Buncic1 and Milorad Mirilovic2
1Department of Veterinary Medicine, University of Novi Sad, Trg D. Obradovica 8, 21000
Novi Sad, Serbia; buncic_sava@hotmail.com
2Department of Veterinary Economics and Statistics, Faculty of Veterinary Medicine, Bulevar
Oslobodjenja 18, 11000 Belgrade, Serbia
Summary
Among a large number of susceptible animal species, the domestic pig is the most important
source of human Trichinella infection worldwide. Trichinellosis is commonly defined by
two cycles; ‘domestic’ (in pigs on-farm) and ‘sylvatic’ (in wildlife). Under conditions on
industrial farms (particularly indoor) with good hygienic practices and efficient management
including biosecurity, combined with effective governmental/veterinary services, Trichinella
transmission via the domestic cycle is unlikely. In countries effectively implementing these
strategies and with officially recognised negligible risk of Trichinella in domestic pigs, testing
for this parasite at meat inspection is no longer mandatory for slaughter pigs reared in
integrated production. In other countries, testing for Trichinella of slaughtered pigs is a very
important component of the control system. Under conditions on small farms with pigs
having access to the outdoors and where control measures are poorly implemented, or are
lacking, the domestic cycle can play a very important role in trichinellosis transmission. This
possibility is further enhanced where socio-economic and political problems temporarily
diminish the efficacy of the governmental/veterinary services. Additionally, the practice of
making uncooked products from meats of uninspected domestic and/or wild pigs at home
represents a major risk for human infection. The risk of further spreading trichinellosis is
additionally exacerbated by increased globalisation in modern times, including increased
movements of livestock, food and people. Traditional farming practices facilitating a mixture
of domestic and sylvatic cycles of Trichinella need to be modified/improved so to ensure the
separation of the cycles. Furthermore, hunters need to be educated to avoid leaving animal
carcasses or their entrails in the field because this increases the probability of transmission
to new hosts. Also, the farmers, the hunters and the consumers should be educated to freeze
pork (including meat from wild boars) before its further home-processing into products, or
to cook the product before consumption, or both, aimed at the larvae inactivation.
Keywords: Trichinella, wild boar, pig, meat, meat products
1. Introduction
Trichinellosis is an infectious parasitic disease transmissible from animals to man (zoonoses).

Natural Trichinella infections in more than 100 species of mammals, seven avian species, and
three reptile species have been reported (Pozio, 2005). Nevertheless, the domestic pig is the

most important source of human infection worldwide. In addition, meats of wild boars and
horses have played a significant role during outbreaks over the past few decades. Trichinellosis
is commonly defined by two cycles; ‘domestic’ and ‘sylvatic’.
With the ‘domestic cycle’, trichinellosis is considered in domestic pigs that can become
infected through feeding on: (a) uncooked swill and other organic wastes that can contain
pork; (b) carrion that are not removed/disposed; or (c) synanthropic animals from the
surroundings (e.g. rodents). Under conditions on industrial farms (particularly indoor) with
good hygienic practices and efficient management including biosecurity measures, Trichinella
transmission via the domestic cycle is unlikely, as indicated by the lack of reports of infections
on industrialised farms in developed, western countries (EFSA, 2005a). However, under
conditions on small farms with pigs having access to the outdoors, and where control measures
are poorly implemented or are lacking, the domestic cycle can play a very important role in
trichinellosis transmission. This possibility is further enhanced where socio-economic and
political problems temporarily diminish the efficacy of the governmental/veterinary services.
With the ‘sylvatic cycle’, trichinellosis is considered in wildlife hosts comprising a number of
carnivorous and omnivorous wild animal species, between which the infection is transmitted
through feeding on: (a) infected prey animals; (b) tissue from infected carrion of the same
species (‘cannibalism’); or (c) tissue from infected carrion of other species. The likelihood
of the transmission is enhanced by the fact that Trichinella larvae can survive in decaying
muscles of dead animals for extended periods of time.
The domestic and sylvatic cycles can function either independently from each other or
interactively (Pozio, 2007). The interaction between the cycles, i.e. a switch from wild animals
to domestic animals, can occur where there is an improper management in segregating
husbandry and wildlife (Gottstein et al., 2009). This usually leads to increases in incidence/
prevalence in susceptible food animals and humans, which is accompanied by serious problems
in international meat trade. The risk of spreading trichinellosis is further exacerbated by
increased globalisation including increased movements of livestock, food and people that is
evident in modern times.
2. Main characteristics of disease
2.1 Animal infection
Trichinellosis is a foodborne parasitic zoonotic disease caused by nematode worms of the

genus Trichinella including T. spiralis T1, T. nativa T2 (and related genotype Trichinella T6),
T. britovi T3 (and related genotypes Trichinella T8, T9), T. pseudospiralis T4, T. nelsoni T7,
T. murrelli T5, T. papuae T10 and T. zimbabwensis T11 (EFSA, 2004). All the species of the
genus are known to infect mammals (mostly carnivores), rodents and omnivores including
pigs, and occasionally also herbivores (e.g. horses), but T. pseudospiralis also can infect birds
and both T. papuae and T. zimbabwensis also can infect reptiles (EFSA, 2004). In pigs in
Europe, T. spiralis is of the highest concern; followed by T. britovi and T. pseudospiralis. Only

Trichinellosis in wild and domestic pigs and public health
the larval stage of the parasite is infectious; the infection occurs only via the ingestion of the
muscle tissue containing the larvae (Figure 1).
However, the infectivity can vary with Trichinella species-animal species combinations (Table
1). Furthermore, both the infective dose and the total number of larvae in muscles can differ
between pigs and wild animals. For example, some reports indicate that in pigs and foxes,
Ingestion of Ingestion of
meat scraps undercooked meat
or animals (esp. pork)
Encysted larva in
striated muscle
Pigs
Carnivorism Carnivorism
Larva released in
small intestine
Rodents
Encysted larva in
striated muscle
Adults in
Circulation small intestine
= Infective stage
Larva deposited
= Diagnostic stage in mucosa
Figure 1. Life cycle of Trichinella (source: http://www.dpd.cdc.gov/dpdx).
Table 1. Trichinella infectivity variations in animals (adapted from Kapel and Gamble, 2000; Trichiporse,
2005; EFSA, 2005b).
Trichinella species Domestic animals Wildlife Birds
Pigs Horses Wild boars Rats Foxes
T. spiralis +++1 +++ +++ +++ +++ -

T. nativa - ? + + +++ -
T. britovi ++ + ++ ++ +++ -
T. pseudospiralis + ++ ++ ++ +++ +
1 Infectivity: +++ highest; + lowest; - negligible.

the infective dose is around 50,000 and 500 larvae, respectively, and average larval burden in
an infected host is around 950 and 20 per host, respectively (Olsen et al., 1964; Kapel et al.,
1995). Normally, infected animals do not show clinical signs of the disease; therefore, in most
EU member states and non-EU European countries, slaughter pigs, horses, wild boar and
other wildlife intended for human consumption are tested for Trichinella at meat inspection.
2.2 Human infection
In contrast to animals, infected humans can develop serious and life-threatening disease.
Historically, most cases of human trichinellosis worldwide have occurred following
consumption of pork. Trichinella infection in humans is strongly associated with the
consumption of raw or undercooked meat; thus, cultural factors such as traditional dishes
based on raw or undercooked meat or meat-derived products play an important role in the
epidemiology of the disease (Gottstein et al., 2009). However, in the last fifty years it has
also become clear that meat from other species – wild boar, bears, foxes, walrus, cougar, dog
and horse – can be a vehicle of trichinellae (EFSA, 2004). Hunters, their relatives and their
friends are at risk of trichinellosis infection when raw meat from game animals is not tested
for Trichinella before consumption. The migratory flow of humans with their own food
practices including the consumption of raw meat, the illegal importation of non-controlled
meat from endemic to non-endemic countries, and new food practices and dishes including
raw meat has resulted in outbreaks in Denmark, Germany, Italy, Spain, and the United
Kingdom (Gottstein et al., 2009).
Trichinella infection with low number of larvae can remain asymptomatic in humans, but in
the case of ingestion of a higher number of larvae (e.g. few hundred), the disease exhibits two
main phases: intestinal and muscular. The intestinal phase is manifested by gastroenteritis
(diarrhea, abdominal pain) approximately two days post-infection, caused by the larvae
penetrating intestinal mucosa and migrating via blood circulation throughout the body until
reaching their final location: the striated skeletal muscles. Migrating Trichinella larvae and
their metabolites provoke an immediate reaction, which causes immunological, pathological,
and metabolic disturbances and the various clinical phenomena observed during the acute
stage of the infection (Gottstein et al., 2009). Human infection data from outbreaks indicate
that some differences in the disease pattern may exist between Trichinella species. For example,
the incubation period was 5-20 days after ingestion of 5,000-18,000 T. spiralis larvae, but 12-40
days after ingestion of 300-30,000 T. britovi larvae (Pozio et al., 1993; Gari-Toussaint et al.,
2004; EFSA, 2005b). In the case of the former species, not only the incubation was shorter,
but the symptoms were also more intense.
The therapy in cases of human trichinellosis must be applied as early as possible, with
application of antihelmintics at the intestinal invasion stage so to eliminate intestinal
forms of Trichinella sp. from the lumen of the gastrointestinal tract. The drugs principally
include preparations like albendazole and mebendazole. In the case of delayed start of the
antihelmintic therapy, i.e. during advanced stage of disease, the effects on already encysted
larvae are poorly elucidated to date. Despite therapy, lethality in cases with high infection
intensity is up to 5%, whilst in milder cases most patients exhibit a disappearance of symptoms
within 2 to 6 months (Gottstein et al., 2009).

3. Current trichinellosis status
3.1 Current European Union perspective
3.1.1 Wild pigs and other wildlife
In the EU in 2007 (EFSA, 2009), the reported Trichinella-positive wildlife animals included
0.1% of non-farmed wild boars (443,890 examined), 4.5% of bears (403 examined), 2.6% of
foxes (6,680 examined), 19.6% of lynxes (224 examined), 19.4% of racoon dogs (222 examined)
and 21.8% of wolves (55 examined).
3.1.2 Domestic pigs
In the EU in 2007 (EFSA, 2009), Trichinella was reported in <0.1% of 220,680,358 examined
domestic pigs (EFSA, 2009) and in 0.4% of 6,615 examined farmed wild boar. The highest
numbers of Trichinella-positive slaughter pigs were reported by Poland, Romania and Spain.
In 2007, Denmark was assigned the status as a region where the risk of Trichinella in domestic
pigs is officially recognised as negligible in accordance with Regulation (EC) No. 2075/2005
(EC, 2005). This was the first time this status was granted to any EU member state. Countries
with this status are allowed to use a risk-based monitoring programme for Trichinella, and
testing for this parasite at meat inspection is no longer mandatory for slaughter pigs reared
under controlled housing conditions in integrated production.
3.1.3 Humans
In the EU in 2007 (EFSA, 2009), a total of 779 confirmed human cases of trichinellosis were
reported. The highest numbers of cases were recorded in Bulgaria, Poland and Romania.
Bulgaria and Romania became EU member states in 2007, thus their contribution has resulted
in a higher number of recorded cases of trichinellosis compared to previous years. In 2007
in the EU, for 69.1% of confirmed human cases, the Trichinella species was not reported, but
where reported, Trichinella spiralis was the most common species (28.2% of all cases). Apart
from T. spiralis, T. nativa and T. pseudospiralis were detected in humans; but no cases due to
T. nativa or T. pseudospiralis were reported.
3.2 Current Serbian perspective
3.2.1 Wild pigs
Trichinellosis is endemic and prevalent in the wildlife in Serbia. This includes the sizeable
wild boar population (roughly 10,000-11,000), of which roughly 25% were hunted annually
during the 2005-2008 period. Whilst it is not known whether every hunted boar was subjected
to examination for Trichinella infection, among annually reported Trichinella tests in hunted
wild boars, between 0.45% and 1.26% were Trichinella-positive (Table 2). On the other hand,
published data for Trichinella in wildlife other than wild boar in the country is scarce.

Table 2. Trichinellosis in wild boars in Serbia.
Year Total number of boars Number of hunted wild boars Positive wild boars: number
(% of tested)
2005 10,781 2,631 14 (0.53)

2006 10,647 2,638 12 (0.45)
2007 10,583 2,531 32 (1.26)
2008 10,839 2,749 27 (0.98)
3.2.2 Domestic pigs
Total numbers of annually reported Trichinella-positive slaughtered domestic pigs in Serbia

during the 1994-2007 period varied between 153 and 1,698 with the incidence ranging roughly
between 0.004% and 0.04% (Table 3). Industrial pig farms usually have well-controlled
farming practices, so have only extremely rarely been involved in trichinellosis. In contrast,
very small farms, most often holding only a few pigs (‘backyard’; often with access to the
outdoors), to be slaughtered on-farm and for home consumption pose the main trichinellosis
risk; this is much higher than that posed by industrial farms. This is supported by data for
2004 (Mirilovic, 2005), showing that the mean number of pigs on positive farms in Serbia
was 13.2, with 40.4% of the positive farms having only 1-7 pigs. The Serbian experience is
that the farm-level epidemiological situation within a given region could be more relevant
Table 3. Trichinellosis in domestic pigs in Serbia.
Year Positive pigs
Number %
1994 153 0.0041

1995 394 0.0094
1996 866 0.0195
1997 1,416 0.0336
1998 1,519 0.0366
1999 1,575 0.0360
2000 1,698 0.0415
2001 1,147 0.0317
2002 920 0.0256
2003 877 0.0241
2004 645 0.0188
2005 391 0.0124
2006 426 0.0113
2007 586 0.0147

for the prevalence of Trichinella in domestic pigs than the geographically defined region
itself (EFSA, 2005b).
When considering the temporal trend of trichinellosis in domestic pigs in Serbia, it is clear that
socio-economic factors can play a major role in the occurrence of trichinellosis in domestic
pigs (Djordjevic et al., 2003; Cuperlovic et al., 2005; EFSA, 2005b). Before the political and
military turmoil in the Balkans, i.e. in 1970s-1980s, pig farming in Serbia was based on very
large state-owned industrial farms, with reasonable farming practices and under very well-
organised veterinary and regulatory activities. At that time, the occurrence of trichinellosis in
domestic pigs was roughly stable, ranging from 0.009% to 0.02% annually (data not shown).
Subsequently, during the decade-long Balkan problems in the 1990s, a sharp decline in large
industrial farms was associated with a sharp increase in very small farms and ‘backyard’
pig rearing. The latter brought about much worsened or sometimes ‘improvised’ farming
practices that were often not subject to proper regulatory controls. This was accompanied
with a correlated increase in trichinellosis occurrence in domestic pigs in the 1990s (Figure 2).
However, since the political and military problems ended and democratic transition occurred
in Serbia in 2000, gradual improvements in farming practices and regulatory activities
took place over the following years, which resulted in a clear decrease of the trichinellosis
occurrence in domestic pigs (Figure 2).
3.2.3 Humans
The total number of reported human cases of trichinellosis in Serbia during the 1994-2007
period varied between 117 and 791, with the incidence ranging roughly between 0.001%
and 0.01%, respectively (Table 4). When considering the temporal trend of trichinellosis in
humans in Serbia, it is clear that socio-economic factors can play a major role in occurrence
of trichinellosis in humans, similar to the situation with domestic pigs (Djordjevic et al., 2003;
Cuperlovic et al., 2005; EFSA, 2005b). In 1970s-1980s, the occurrence of human trichinellosis
was mostly stable, at around 0.5 cases per 100,000 population, with two exceptional peaks
1,600
1,400
1,200
1000
Pigs
800
600
400
200
0
1994 1995 1996 1997 1998 1999 2000 2001 2002 2003
Year
Figure 2. Temporal trend of trichinellosis in pigs in Serbia (Mirilovic, 2005).

Table 4. Trichinellosis in humans in Serbia.
Year Human cases
Total number Number per 100,000 population
1994 492 6.56

1995 791 10.55
1996 594 7.92
1997 766 10.22
1998 408 5.44
1999 559 7.46
2000 411 5.48
2001 383 5.11
2002 575 7.67
2003 178 2.37
2004 221 2.95
2005 339 4.52
2006 188 2.51
2007 117 1.56
of 1.0 due to two larger outbreaks (data not shown). In 1990s, due to the Balkan political
and military problems, the occurrence increased to between 5.4 and 10.5 per 100,000;
subsequently, starting in late 1990s and through 2000s, it significantly decreased following
the democratic and economic transition in the country (Figure 3).
1000
800
Humans
600
400
200
0
1994 1995 1996 1997 1998 1999 2000 2001 2002 2003
Year
Figure 3. Temporal trend of trichinellosis in humans in Serbia (Mirilovic, 2005).

When considering some factors associated with infections in humans, no clear effect of
gender on trichinellosis occurrence has been observed (Table 5). On the other hand, fewer
cases of trichinellosis occurred in young children and older individuals (≤ 6 and ≥ 60 years,
respectively) compared to other age groups (Table 5). The reasons behind this phenomenon
are unclear, but it could be hypothesised that the diets of very young and older contain
somewhat smaller amounts of uncooked meat (pork) products and/or that their digestive
system is somewhat less efficient in digesting meat (so fewer larvae are freed) and/or that
their age groups are smaller in proportion to the total population, compared with other age
groups. In any case, it is clear that the large majority (approximately 70%) of all human cases
occurred in individuals being between 20 and 60 years of age.
3.2.4 Source attribution for human infection
When considering source attribution for human trichinellosis, it is important to bear in

mind that not all larvae in ingested meat are infective. The larvae infectivity, so human dose-
response, are affected by duration and temperature (chilled, frozen) of meat storage before
consumption, salt concentration i.e. water activity (aw) of meat/product and cooking regime
(temperature, time) of meat/product. These factors are highly variable and inter-related,
and so are difficult to evaluate and/or quantify. From a practical perspective, the two most
important meat-associated factors that can completely inactivate the most important species,
T. spiralis, in meat are effective freezing and cooking regimes (Table 6).
With respect to the effects of freezing, T. spiralis and T. pseudospiralis can be considered as
cold sensitive trichinellae when in the muscle of pigs and wild boar (EFSA, 2004). Concerning
the cold resistant Trichinella species/genotypes that have been identified (e.g. T. nativa,
Trichinella T6 and T. britovi), it has been noted that present cold treatment and time/
temperature combinations targeting T. spiralis may not be effective in respect to the cold
resistant strains should they be present in meat of pigs; however, there is a lack of scientific
data on this issue (EFSA, 2004). With respect to the effects of cooking, no information is
available regarding the heat tolerance in meat of species/genotypes of trichinellae other
than T. spiralis. Furthermore, it should be noted that the effectiveness of heat inactivation of
trichinellae can also vary with the meat heating method; a somewhat higher temperature for
Table 5. Age and gender of human trichinellosis cases in Serbia (1994-2003).
Age groups (years) Males (number) Females (number) Total
Number %
≤6 101 103 204 3.96

7-19 518 411 929 18.01
20-39 1,102 866 1,968 38.16
40-59 930 700 1,630 31.61
≥60 210 216 426 8.26
All age groups 2,861 2,296 5,157 100.00

Table 6. Inactivation of Trichinella spiralis by freezing and cooking (adapted from EFSA, 2004).
Meat freezing inactivation of T. spiralis Meat cooking inactivation of T. spiralis
Core temperature (°C) Required duration Core temperature (°C) Required duration
-17.8 106 hours 49.0 21.0 hours

-20.6 82 hours 50.0 9.5 hours
-23.3 63 hours 52.2 2.0 hours
-26.1 48 hours 54.5 30 minutes
-34.5 8 hours 60.0 1 minute
-37.2 0.5 hours 62.2 instantaneous
complete inactivation may be required in the case of microwave-oven heating, compared to

conventional-oven heating (EFSA, 2004).
Regarding source attribution for the human cases in Serbia, the published data is limited
and poorly documented, which does not allow detailed related analysis. Nevertheless, it is
considered that no confirmed human cases have been linked to meat/pork from industrial
abattoirs to date. On the other hand, anecdotal evidence from most human trichinellosis cases
points to home-made traditional meat products that are cured, cold smoked and dried but
not subjected to any heat treatment either during the production or before the consumption,
as the sources of infection. The two main examples of those meat products are country-style
dried meats and fermented, dry sausages (salamis).
In home-making of cured, cold smoked and dried pork joints (ham, shoulder), the production
parameters vary and are largely undocumented but, based on the epidemiological information,
Trichinella larvae can survive in such products if they contain infected meat. For illustration
purposes, typical parameters in industrial production of corresponding dried meats are: rapid
chilling of pork (0 °C after 24 h; pH 5.8), salting at 5 °C (until aw ~ 0.96, i.e. 4.5% NaCl), cold
smoking (≤22 °C), drying (≤15 °C) and maturation (12-18 °C; relative air humidity 70-78%).
In home-making of cured, fermented, cold smoked and dried sausages (salamis), the production
parameters also vary and are largely undocumented but, based on the epidemiological
information, Trichinella larvae can survive in such products if they contain infected meat. For
illustration purposes, typical parameters in industrial production of corresponding sausages
are: rapid chilling of pork (0 °C after 24 h; pH 5.8), chopping-mixing of ingredients (55-70%
lean pork, 25-40% fatty tissue, 3% curing salts (NaCl, nitrate and nitrite), 0.5% spices and
flavouring), stuffing into casings, fermentation (15-40 °C; 2-5 days), cold-smoking (few hours)
and drying to different extents (1-4 weeks for semidry sausages; 12-14 weeks for dry sausages).

Overall, the common scenario leading to human trichinellosis in Serbia involves consumption
of home-made meat products that did not receive heat treatments (mentioned above) but
which contained:
• meat from a domestic pig slaughtered under home arrangements and not subjected to
post mortem veterinary inspection; or
• mixture of meats from multiple domestic pigs among which some were post mortem
veterinary inspected but one was slaughtered under home arrangements and not subjected
to post mortem veterinary inspection; or
• meat from a hunted wild boar not subjected to post mortem veterinary inspection; or
• mixture of meats from domestic pig and wild boar, where one (or both) of them were not
subjected to post mortem veterinary inspection.
Obviously, the most relevant aspects of the scenario are: domestic slaughter/hunting, lack
of meat examination for Trichinella, lack of heat treatment and, when available, use of wild
boar meat.
4. Main principles of Trichinella controls
4.1 Global controls in wildlife
The risk caused by the wildlife reservoir is dependent on the contacts between domestic pig
and wildlife. When only minimal contacts with wild omnivores/carnivores exist, as in the
case of industrialised farms, the importance of the wildlife reservoir is certainly smaller than
in the case of more traditional farming systems (EFSA, 2005a). With respect to surveillance of
wildlife reservoirs, it is usually based on collecting and testing foxes from the area. However,
there are large variations in Trichinella prevalence in wildlife even within a single country.
Furthermore, due to mobility of wildlife and other variable factors, the geographical area to
which one wishes to make inferences on the prevalence in wildlife based on wildlife surveys
is difficult to define (EFSA, 2005a). Overall, monitoring of a wildlife reservoir can be helpful
in assessing its potential as source for Trichinella exposures of domestic pigs, but there are
difficulties in using wildlife monitoring to verify the absence of Trichinella in wildlife.
It appears, that the only measure that can be implemented to reduce the prevalence of infection
among wildlife is to instruct hunters to avoid leaving animal carcasses and/or entrails in the
field, a practice which increases the probability of transmission to new hosts (Gottstein et
al., 2009). Namely, a very close relationship between the conventional hunter’s practice to
leave animal carcasses in the field after skinning and the prevalence of trichinellosis among
wildlife has been documented in arctic, subarctic and some European regions. Furthermore,
continuous rodent control in the areas/locations enabling domestic pig exposure is another
very important aspect of Trichinella reservoir control.
4.2 Global controls in domestic pigs
Control of Trichinella infection in pork has traditionally been accomplished by inspection

of individual carcasses at slaughter or by post-slaughter processing to inactivate parasites.

The changes in animal husbandry on industrial, indoor farms have virtually eradicated the
domestic Trichinella cycle from many parts of Europe. However, in many of the new EU
Member States and the EU candidate/associated countries (such as Serbia), more traditional
farming practices facilitating a mixture of domestic and sylvatic cycles of Trichinella may
be more common; the mixture represents the major infection risks to domestic pigs (EFSA,
2005a). This is indirectly confirmed by the fact that the Trichinella-infected pigs in the EU
nowadays are primarily from backyard, free-ranging, organic or small family farm production
systems (EFSA, 2005a). Therefore, the main strategies of Trichinella controls in domestic pigs
are aimed at effective separation of the domestic and sylvatic cycles.
Declines in the prevalences of this parasite in domestic pigs in developed countries during
the last 30 years, combined with improvements in pork production systems, indicated that
it is possible to ensure pork safety in respect to Trichinella at the farm level (Gajadhar et al.,
2009). In practice, related measures including aspects of farm management, bio-security, feed
and feed storage, rodent control programs, and general hygiene are incorporated in the good
farming practices system, details of which are described in an EU scientific opinion (SCVPH,
2001) and Regulation (EC) 2075/2005 (EC, 2005). Strict application of those measures ensuring
effective separation of sylvatic and domestic cycles, combined with regulatory-documented
audits of the system and adequate on-farm monitoring of the parasite, makes achieving the
Trichinella-free farm status possible. In countries effectively implementing this strategy and
with an officially recognised negligible risk of Trichinella in domestic pigs, testing for this
parasite at meat inspection is no longer mandatory for slaughter pigs reared under controlled
housing conditions in integrated production. In other countries, testing for Trichinella of
slaughtered pigs is a very important component of the control system.
4.3 Global post-harvest controls
Pork from pigs that have not been tested for Trichinella – for whatever reason – should be
preferably processed commercially, or at least treated by the consumer, using methods proven
to inactivate the parasite. As required by many regulatory authorities, ready-to-eat pork
products such as dried meats and cold-smoked sausages must be processed to kill Trichinella
larvae by heating, cooking, or curing (EFSA, 2004; Gajadhar et al., 2009). This strategy is of
particular interest in countries with endemic trichinellosis and higher proportion of small-
scale (‘backyard’) pig farming. In those countries, the situation is often worsened by frequent
preparation of uncooked meat products under domestic (uncontrolled) arrangements, using
meat from uninspected ‘backyard’ pigs that can be mixed with hunted wild boar meat when
available. Although all feasible efforts should be made to discourage the ‘home-making of
products from uninspected pork’, various negative socio-economic factors can make their
total elimination in the short-to-medium term unrealistic in those countries including Serbia.
As an important measure in the meantime, the farmers, the hunters and the consumers
should be educated and advised to freeze the pork (including from wild boars) before its
further home-processing into products, or to cook the product before consumption, or both.
With respect to home-made fermented, dry sausages (salamis) where cooking either before
or after preparation is traditionally unacceptable for reasons of sensory quality, but where the
Trichinella-inactivation effects of the curing are unreliable, the advice to keep the sausages
in the freezer for some time before consumption would be beneficial.

5. Conclusions
Natural Trichinella infections in more than 100 species of mammals, seven avian species, and
three reptile species have been reported, but the domestic pig is the most important source of
human infection worldwide. Trichinellosis is commonly defined by two cycles, ‘domestic’ (in
pigs on-farm) and ‘sylvatic’ (in wildlife). Under conditions on industrial farms (particularly
indoor) with good hygienic practices and efficient management including biosecurity, and
combined with effective governmental/veterinary services, Trichinella transmission via the
domestic cycle is unlikely, as indicated by the lack of reports of infections on industrialised
farms in developed, western countries. In countries effectively implementing these strategies
and with an officially recognised negligible risk of Trichinella in domestic pigs, testing for this
parasite at meat inspection is no longer mandatory for slaughter pigs reared under controlled
housing conditions in integrated production. In other countries, testing for Trichinella of
slaughtered pigs is a very important component of the control system.
A switch of trichinellosis from wild animals to domestic animals can occur where there is
improper management in segregating husbandry and wildlife. Under conditions on small farms
with pigs having access to the outdoors and where control measures are poorly implemented
or are lacking, the domestic cycle can play a very important role in trichinellosis transmission.
This possibility is further enhanced where socio-economic and political problems temporarily
diminish the efficacy of the governmental/veterinary services. Additionally, the practice of
making uncooked products from meats of uninspected domestic and/or wild pigs at home
represents a major risk for human infection. These problems usually lead to increases in
incidence/prevalence in susceptible food animals and humans, as illustrated by the Serbian
situation in 1990s. The risk of further spreading trichinellosis is additionally exacerbated by
increased globalisation including increased movements of livestock, food and people that is
evident in modern time.
Traditional farming practices facilitating a mixture of domestic and sylvatic cycles of

Trichinella need to be either modified/improved so to ensure that cycles are separated or
gradually totally replaced; however, achieving the latter is not realistic in the short-to-
medium term because of traditions and practical difficulties. Furthermore, hunters need
to be educated to avoid leaving animal carcasses and/or entrails in the field because this
increases the probability of trichinellosis spread to new hosts. Also, the farmers, the hunters
and the consumers should be educated and advised to freeze the pork (including meat from
wild boars) before its further home-processing into products, or to cook the product before
consumption, or both, aimed at the larvae inactivation.

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Influence of climate change on diseases of wild
animals
Armin Deutz1, Thomas Guggenberger2 and Johann Gasteiner2
1Styrian Provincial Government, Bahnhofviertel 7, 8850 Murau, Austria;
armin.deutz@stmk.gv.at
2Federal Agricultural Research and Education Centre Raumberg-Gumpenstein, Raumberg
38, 8952 Irdning
Summary
The occurrence of transferable diseases in wild animals generally reflects the causative agent’s
distribution. The influence of climate change on the prevalence of pathogenic organisms is
either exerted directly by causative agents with an increased survival rate as related to higher
average annual temperatures, or indirectly because vectors (e.g. ticks, mosquitoes) experience
better environmental conditions similarly promoting their viability and allowing them to
invade new areas of circulation. The latter option explains why pathogen-carrying vectors
such as ticks and mosquitoes prevail at higher sea levels than observed two decades ago.
In recent years, vectors with seasonal cycles show longer periods of incidence. In addition,
parasitic eggs, larvae and parasitic organisms at intermediate stages can be found at higher
sea levels as they too benefit from changing climate conditions. For this reason cumulative
purulent and abscess forming pneumonia is observed in chamois which originate from
lungworm infections. This contribution demonstrates the relationship between climate factors
and transferable diseases. The geographical dissemination and incidence of many diseases
interferes with environmental changes. Additionally, risk factors increasing the diseases’
prevalence and incidence are discussed. Finally, selected examples of our own findings on
the impact of climate changes on diseases (tularaemia) and on habitats of alpine wild animal
species (black grouse, snow grouse, chamois and ibex) are highlighted.
Keywords: climate warming, disease risks, Tularaemia, habitats
1. Introduction
Wild animals as well as infectious agents and vectors/reservoirs are greatly influenced by
climate, geographical location, structure, fauna and flora and they may migrate deep into
human settlement areas, depending on animal species. There are several examples for the
impact of climate change on enzootic infections. Whilst this may be in accordance with
ecological requirements, it also poses the danger of animal-human disease transmission.
The spread of particular bacterial, viral and helminthic infections (partly with zoonotic
potential) is influenced by factors of climate change, especially temperature and distribution
of precipitation (Williams et al., 2002; Hoberg et al., 2008).

The following variables and parameters related to climate change represent significant risk
factors as regards wild animal diseases:
• influence on infectious agents (basic reproductive number, tenacity, etc.);
• increase of populations (e.g. wild boar, vectors, etc.);
• increase of temperature especially in alpine regions;
• decrease of habitat quality for alpine animal species (immunosuppression) with increased
host susceptibility;
• heat waves, heat stress, water shortage;
• hygienic problems in feeding of wild animal species (red deer, row deer, wild boar),
particularly at warm ambient air temperature (e.g. mild winter) and especially when
feedstuff is presented on the ground;
• ‘new’ infectious agents (e.g. West Nile virus, hepatitis E virus, Hanta viruses, EHEC);
• changes in habitats (e.g. barrier lakes);
• global trade with animals and food of animally origin;
• increase of host range of some infectious agents?
• illegal international pick up of stray dogs and cats;
• international transport and movement of wild animals and of straying animals;
• increasing international travelling of humans.
Climate change influences the dissemination and the reproduction of both pathogenic
organisms and of wild animals (Figure 1).
There is already a widespread change in natural calendars (phenology) of plants and animals,
as well as change in some species distributions. Now so-called threshold changes (i.e. sudden,
fundamental changes) in ecosystems are beginning to be observed in nature. This suggests a
Wild Animal Infectious Agent

genetics, population Contact tenacity,
density, age infectivity,
distribution, RISK OF infection dose,
sex ratio, resistance,
DISEASE
…. ….
Stress Trans-
mission
Environment
habitat, climate (change),
tourism, hunting, ...
Figure 1. Interactions between wildlife, infectious agents and environmental factors (Deutz and
Gressmann, 2001).

Influence of climate change on diseases of wild animals
future with nature and ecosystems very much in flux, which will have major epidemiological
implications (Lovejoy, 2008).
Among wild animals some species will benefit from climate change by increasing their
population (e.g. wild boar) while other species may diminish or become extinct. These
‘underdogs of climate change’ (e.g. chamois, ibex, black grouse and snow grouse) would find
themselves in suboptimal habitats whilst the infectious pressure is increasing. Additionally, it
needs to be considered that the number of ‘new’ emerging pathogenic agents, predominantly
zoonoses, occurring in Middle Europe is increasing.
Most vector-borne diseases exhibit a distinct seasonal pattern, which suggests that they are
weather sensitive. Precipitation (rainfall), temperature and other weather variables in many
ways affect both the vectors and the pathogens that these transmit. High temperatures can
increase or reduce survival rate, depending on the vector, its behaviour, ecology and other
factors. Thus, the probability of transmission may be increased by higher temperatures.
Moreover, the tremendous growth in international travel increases the risk of importation of
vector-borne diseases, some of which can be transmitted locally under suitable circumstances
and can become autochthonous (Gubler et al., 2001).
The complex interaction of factors like environmental and ecological changes, social factors,
decline of health care, human demographics and behaviour influences the re-emergence of
such diseases. Viruses, especially RNA viruses with their ability to adapt quickly to changing
environmental conditions, are among the most prominent examples of emerging pathogens
(Ludwig et al., 2003). The effects of climate and population movements and other risk factors
on emerging infectious diseases in men, domesticated animal as well as in wildlife need
to be seriously reconsidered. Overall, the success of disease control measures has been
disappointing. Scientists must accept the challenge of communicating effectively with the
public and policy makers worldwide if success is to be achieved (Fayer, 2000).
Vector-borne diseases gained importance in men as well in animals in Middle Europe during
the past years. Reports on severe diseases, formerly only known as so called ‘travel sickness’
from tropical countries, markedly increased in number in the recent years (Zinstag and
Schelling, 2003; Ready, 2008; Schwaiger and Bauer, 2009). Climate models predict a global
warming of 1.4 °C up to 5.8 °C until the year 2100 and especially arthropod-borne diseases
are strongly influenced by the climate. Schwaiger and Bauer (2009) listed arthropod-borne
pathogens with their reservoirs, which are relevant for Germany, and most of which have
zoonotic potential (Table 1).
In a screening for West Nile virus in 2003, all samples from free-living birds and horses from
Austria were negative for WNV (Weissenböck et al., 2003). In 2008 the first cases of West Nile
virus infections in free-living birds, especially goshawks (Accipiter gentilis), were detected
in Austria. Interventions to control West Nile virus include mosquito and bird population
surveillance (Epstein, 2001).
Already in the year 1974 Thiel postulated that Q fever was found in both hemispheres
within the 10 °C annual isotherms, especially where the climate was warm and dry as in the

Table 1. Arthropod-borne pathogens and their reservoir in Germany (Schwaiger and Bauer, 2009).
Viruses Bacteria Parasites
Tick-borne encephalitis virus Borrelia burgdorferi s.l. Babesia spp.

West Nile virus Ehrlichia chaffeensis Leishmania spp.
Bluetongue virus Anaplasma phagozytophilum Hepatozoon canis
Batai virus Ehrlichia canis Dirofilaria spp.
Tahyna virus Rickettsia colorii-complex
Uukuniemi virus Rickettsia slovaca
Eyach virus Rickettsia sibirica
Tribec virus Rickettsia massiliae
Lipovnik virus Rickettsia helvetica
Bahanja virus Rickettsia aeschlimanii
Semliki Forest complex virus Rickettsia akari
Lednice virus Rickettsia felis
Sindbis virus Rickettsia monacensis
Rickettsia typhi
Coxiella burnetii
Francisella tularensis
Bartonella quintana
Mediterranean and Black Sea areas, the steppes of central Asia, the African savannas and
the grassland areas of North America and Australia (Thiel, 1974). In the last few years, Q
fever cases have occurred increasingly in Middle and Northern Europe and an influence of
climate change is discussed. Predisposed groups (e.g. veterinarians, farmers, slaughterhouse
workers, hunters) have to recognise zoonotic risks and observe prophylactic measures (Deutz
et al., 2001, 2003).
A recent increase in the occurrence of Dirofilaria repens and Dirofilaria immitis has been
reported from Slovakia and Hungary. In September 2007, the first autochtonous case of canine
Dirofilaria repens in a dog was diagnosed in Austria (Duscher et al., 2009; Löwenstein and
Spallinger, 2009). Wild carnivores could also be a reservoir for Dirofilaria spp. Temperature
is one of the main factors preventing the spread of leishmaniasis into Northern Europe.
Evidence indicates that Leishmania infantum is prevalent only within the 5-10 °C January and
20-30 °C July isotherm. Human disease models predicted a dramatic increase in the incidence
of visceral leishmaniasis and a slight increase in the incidence of cutaneous leishmaniasis.
Climate change would most likely also effectuate parasite development and the infection rate
in dogs and thus overall disease incidence (Kuhn, 1999).
Climate variables are known to affect the prevalence, intensity and regional/geographical
distribution of parasites, influencing free-living larval stages or influencing invertebrate as
well as vertebrate hosts. The impact of climate change appears to be more pronounced in

trematodes, and is shown by increased cercarial production (Fayer, 2000; Mas-Coma et al.,
2008, 2009).
Parasitic eggs, larvae and parasitic organisms at intermediate stages were also found at higher
sea levels as these species also benefit from changing climate conditions. As average annual
temperatures in alpine space increase, the time of development and reproduction cycles of
parasites is reduced and therefore more generations of parasites per year may occur. The
cumulative findings of purulent and abscess forming pneumonia in chamois as a result of
lungworm infections can be seen as a direct consequence of these changes (Schaumberger et
al., 2006; Prosl, 2008). As regards trichinellosis there is a hypothesis that the distribution of
T. nativa and T. britovi relates to the environmental temperature, notably the isotherm -4 °C
and -5 °C isotherm in January, respectively (Pozio et al., 1998).
During the hot summer of 2003, an enormous heat stress in both farm animals and wild
animals was triggered in Austria. Especially territorial living wild ruminants (e.g. roe deer)
had severe problems to get to adequate places for their water supply. Consequently, average
body weights in shot wild ruminants, were markedly reduced during this year. A higher
susceptibility for diseases (e.g. paratuberculosis, endoparasites) is likely to ensue. Climate
scientists predict more hot summers and higher numbers of hot days per summer (Kromp-
Kolb and Formayer 2005). It is also possible, that potential vectors (e.g. specific ticks) will also
suffer from hot weather conditions whilst others, particularly mosquitoes, may benefit from
higher temperatures. As a result, some vectors (to date not occurring in Middle Europe) will
encounter environmental conditions that are essential for their life and reproduction. As an
actual example, bluetongue disease, an acute and epidemic infectious disease in sheep, goats
and wild ruminants raged in many European countries (Purse et al., 2008).
In the following two sections selected examples of our own findings (Schaumberger et al.,
2006; Deutz et al., 2009) concerning the influence of climate change on diseases of wild
animals and on habitats of alpine are discussed.
2. Tularaemia in Austria
Bacterial infectious diseases, such as tularaemia, are strongly related to climate parameters
as the pathogen requires specific ambient conditions. Francisella tularensis is a cold-resistant
bacterium that is able to reproduce at moderate temperatures but will not survive at elevated
temperatures. In addition, the population density of host animals and the abundance
of potential disease vectors (ticks, gnats) also play a decisive role in the epidemiology of
tularensis. The infection is transmitted to animals and humans by direct contact with infected
animals and vectors or by inhalation of pathogens or the consumption of insufficiently cooked
hare meat (Ellis et al., 2002). Several cases of tularaemia are recorded in Austria each year,
but the number of unreported cases is likely to be markedly higher. Francisella tularensis is
also considered as a potential biological weapon due to its low infection dose.
We aimed at investigating the impact of climate and weather on the prevalence of tularaemia
in hare populations in the lowlands of eastern Austria.

A total of 271 cases of tularaemia in hares have been recorded in the area under investigation
(Lower Austria, Burgenland, and Styria) in the period from 1994 to 2005, all of which were
geo-referenced according to sender postal code. Temperature and precipitation data for the
selected region were available from 30 meteorological stations of the Central Institute for
Meteorology and Geodynamics in Vienna (Auer et al., 2001). These data provided the basis
for calculating an altitude dependent temperature distribution for suitable monthly mean
values and period sums. The spatial distribution of precipitation was calculated using the
geo-statistical universal kriging method without taking into account the influence of altitude.
These data were used for a two-step analysis. The first step led to boundary values for the
spatial distribution of tularaemia. These boundaries were used to estimate the distribution
of tularaemia in the year 2035. The second step explained the different annual incidences
using actual climate data.
4. Results
4.1 First step: finding spatial boundary values and estimation of the areal
distribution of tularaemia until 2035
A high incidence probability, based on the local isoline encircling the study area, was obtained
for annual precipitation totals below 720 mm, summer precipitation below 180 mm, winter
temperatures above 0.5 °C and mean May temperatures above 14 °C. These limit values
allowed a calculation of the diseases´ spatial distribution for current and future conditions. A
warming of 2 to 4 °C having been induced by climate change was assumed for predicting the
distribution area of the disease in 2035, with warming expected to be more intensive in higher
altitudes than in lowlands. Figure 2 shows the possible spatial distribution of tularaemia in
2035 following a rise in mean annual temperatures. Precipitation was not taken into account
2005
2035
0 25 50 100 150
Figure 2. Possible spatial distribution of tularaemia in Austria in 2035 following a rise in mean annual
temperatures.

due to the lack of a suitable scenario. Under these conditions, tularaemia will slowly spread
from the eastern lowlands via the Danube valley to the west and southern Styria proceeding to
the south. Additional incidents of the disease could also occur in inner-alpine areas providing
favourable climatic conditions.
4.2 Second step: explaining coherences between climate parameters and

incidence
Inside tularaemia zones a clear correlation between the two climate parameters and local
disease incidence was established, which can be represented by the following linear regression
model:
Number of cases per year = 52.12 + 4.08 x (average of monthly mean temperatures for
December, January and February) – 3.46 x (monthly mean temperature for May) + 0.26 x
(precipitation total for June and July)
This formula does not allow calculating absolute numbers of incidence in nature, because it
is based on sample data of one specific region. Particularly noteworthy, however, is the highly
significant (P<0.05) influence of the parameters selected of the incidence rate of the disease
and the coefficient of determination obtained (R 2 = 74.6%). It becomes clear that about ¾ of
inter-year differences can be explained by temperature and precipitation conditions: warm
winter temperatures result in an increase in incidence, while warm May temperatures lead to
a decrease; high precipitation in summer has an increasing effect again. The ideal conditions
for the spread of the disease are thus warm winters combined with low temperatures in May
and high precipitation in summer (Figure 3). The result represents a feasible development
of hares. Warm winter increases the population of hares. Low May temperatures and wet
summers degrade the leverets. Thus, bacteria causing tularaemia find better conditions to
reproduce. This correlation has been derived from observations and obviously does not apply
to arbitrary temperature and precipitation values.
These findings provided the basis for specifying empirical limits for the parameters defined
in the formula, which best correspond to the actual spatial distribution obtained by
geographical analysis. Hence, the probability of tularaemia occurrence is high for a total
annual precipitation below 720 mm, a summer precipitation rate around 180 mm, a winter
temperature above 0.5 °C and a May temperature below 14 °C.
A temperature increase of 2 to 4 °C was assumed for predicting the distribution area of the
disease by 2035 (Figure 4). Under these conditions, tularaemia might slowly spread from the
eastern lowlands via the Danube valley to the west and via southern Styria further to the
south. Additional incidents of the disease could also occur in inner Alpine areas providing
favourable climatic conditions. This means that an extension of the potential tularaemia
distribution area (from currently 13 to 46.5% of the Austrian territory; Figure 2) can be
expected.

mean temperature winter + 0.5 °C

mean temperature May +14 °C
annual precipitation Tularaemia high
precipitation June - July 180 mm incidence probability:
disease incedence
no cumulation ~ winter temperature ≥ + 0.5 °C
border area of cumulation ~ temperature in May < 14 °C
maximum of cumulation ~ summer precipitation > 180 mm
~ annual precipitation < 720 mm
Figure 3. Isolines of analysis and high tularaemia incidence probability theory.
Mean temperature
degree Celsius
4
2
Figure 4. Estimate of future warming in Austria over the next 50 years.

5. Loss of habitats in Alpine regions
Wild animal species such as the black grouse, snow grouse, chamois and ibex have adapted
to life in an Alpine environment above the tree line in the course of their evolution and
thus form part of this very sensitive ecosystem. The habitat of these wild animal species
might be substantially reduced by a potential upward movement of the tree line because of
climate change. An attempt was made to quantify these changes with the aid of models and
a geographical information system (GIS). The calculations were based on the temperature
development of the past 50 years and an estimate of future warming. The scenario derived
from the climate model used predicts a warming of approximately 2.2 °C for the area under
investigation (Niedere Tauern, province of Styria, Austria) over the next 50 years (Figure 5).
The elevation of the tree line strongly depends on the temperature and a high correlation
was found between the growth limit of trees and the 10 °C July isotherm (Daubenmire,
1954; Böhm et al., 2001; Grace et al., 2002; Holtmeier, 2003; Gobiet et al., 2006). The climate
model shows that the relevant isotherms will rise by approx. 450 m over the next 50 years.
The temperature changes, however, strongly depend on the climate model used. Without
additional research work, no statement can be made on how fast the tree line is advancing
towards the temperature related growth limit. Human management practices also have a
substantial influence on tree line position so that future changes in the tree line cannot be
derived from climatic data alone.
When accepting the theory of an elevated tree line due to climate warming, the habitat of wild
animal species will be diminished massively (Figure 6 and 7). For individual species there
are almost no possibilities to avoid or to cross over to other habitats. Taking into account
suboptimal habitats left, disintegration of biotopes, overexploitation of remaining alpine
areas due to touristic and sportive leisure time behaviour by people, endangered species will
Figure 5. Area of investigation (Niedere Tauern, province of Styria, Austria).

Figure 6. Elevation of the tree line with loss of habitats for alpine wild animal species.
Tree line 2040-2050: 2,415 m
Tree line 1950-1960: 1,970 m
6.9 May - October isotherm decade 1950-1960 and decade 2040-2050
Figure 7. Tree line on base May-October isotherm decade 1950-1960 is about on 1,970 m and tree line on
estimated isotherm decade 2040-2050 will be on 2,415 m.
diminish strongly. As a consequence, inbreeding will increase, genetic resources will become
poor, individual immune system will weaken and parasitic and stress-related diseases but
also infectious diseases will emerge.

Several populations will diminish under a critical size of their population or herds and
therefore some species may become extinct. As examinations in Bighorn-Sheep (Ovis
canadensis) showed, populations with less than 50 individuals became extinct within 50
years. Populations with more than 100 individuals were able to survive. Therefore, we can
assume that at least 50 individual animals are necessary to maintain genetic variability, but
for long ranging survival at least, 500 individuals should prevail. Following this approach
only the effective size of the population must be considered, which means that only animals
able to reproduce must be taken into account.
The current habitats of the animal species investigated were determined and mapped using
knowledge based habitat model and a GIS. As an example, the present suitable habitats for
snow grouse are shown in Figure 8. Assuming that the future tree line will adjust to the
changed temperature of the decade 2040-2050, this shift will lead to a dramatic loss in suitable
habitats, which may range from 78 to 98% depending on the season and animal species. The
habitat loss to be expected for snow grouse is illustrated in Figures 8 and 9.
The present study shows, that the losses of convenient wildlife habitats is considerable for all
of the four observed wildlife species. Smaller populations and flocks of these species will not
survive due to loss of their living space, increasing susceptibility for diseases, rising individual
losses caused by predators due to reduced clearness of habitats (higher vegetation). Most
dramatic losses of habitats can be seen for the black grouse. When accepting the modelled
Suitability
temporary possible
minor
good
Figure 8. Current (2010) habitat suitability for snow grouse (Lagopus mutus helveticus).
Suitability
temporary possible
minor
good
Figure 9. Predicted habitat (decade 2040 to 2050) suitability for snow grouse (Lagopus mutus helveticus)
assuming a temperature increase of 2.2 °C and derived shift in the tree line.

scenario for the region ‘Niedere Tauern’, black grouse are in danger of extinction. Snow grouse
may adapt to treeless but cragged alpine areas and therefore should be able to survive. But also
populations of snow grouse would decrease. Chamois and ibex are able to migrate to woody
regions and then problems with damage of trees are likely to occur.
Living in suboptimal habitats is a problem for chamois (Figure 10 and 11) and ibex as
populations might diminish. Infectious diseases like scabies and ablepsia of the chamois, but
also endoparasites would reduce the populations. Smaller groups of these wild animals would
show extended rutting season and then bucks would be weakened. Then, some populations
will be too small and genetic exhausting would be reason for inbreeding depression and for
their extinction. As an example for an epidemic spread caused by changing climate conditions
the outbreak of Infectious ceratoconjunctivitis in chamois (Figure 12) is presented. This
outbreak occurred in 2006 and more than 80 cases of diseased animals where seen in the
districts Murau, Judenburg and Liezen. In 2006, with a very mild autumn, vectors of the
disease (flies) could be observed in alpine regions at the beginning of December 2006. It can
be concluded that the time span, during which an infection is possible, is prolonged.
Suitability
temporary possible
minor
good
Figure 10. Current (2010) habitat suitability for chamois (Rupicapra rupicapra).
Suitability
temporary possible
minor
good
Figure 11. Predicted habitat (decade 2040 to 2050) suitability for chamois (Rupicapra rupicapra) assuming
a temperature increase of 2.2 °C and derived shift in the tree line.

Figure 12. Infectious ceratoconjunctivitis, Chamois.
6. Conclusions
It has been established that global changes, including an increase in trade and global warming,
which act on the environment, are likely to influence the evolution of pathogens and hence
of diseases. To anticipate the risks created by this new situation, two methods have been
developed (Dufour et al., 2008): the first step is to indentify the diseases the incidence or
geographical distribution of which could be affected by global warming, and the second step
is to evaluate the risk of each of these diseases. Further recommendations were to develop
epidemiological surveillance, to increase knowledge of epidemiological cycles, to develop
research concerning these diseases and to pool cross-border efforts to control them.
In summary it can be stated that climate is the key parameter for explaining the distribution
of tularaemia in the past and that limit values for the individual climate parameters can
be identified. The expected warming could result in a massive expansion of the potential
tularaemia distribution area. Yet, to date, little if any attention has been paid to informing risk
groups (hunters, foresters, farmers, laboratory staff, taxidermists, housewives, etc.) beyond
current prevalence regions and recommending them to take preventative measures of working
hygiene (protective gloves, moistening the fur when skinning hares, insect protection, face
masks in the lab) when handling hares and rodents, and following good kitchen hygiene
practices when preparing and cooking hares.

It is a matter of urgency, that veterinarians, physicians, wild life biologists and epidemiologists
consider new emerging diseases. Surveys among wild animal species should be enforced and
serological databases should be developed. Thus, changes in incidences could be detected
earlier and affected regions but also time spans of outbreaks could be pre-estimated. The
knowledge and the vigilance concerning new diseases must be promoted by constant and
up to date information including all involved persons. In addition, examination centres and
laboratories have to be watchful that the spectrum of pathogenic agents will change. West Nile
Virus, Usutu Virus or Louping Ill, Hepatitis E, Krim-Kongo-Fever and Ehrlichiosis display
only a few examples for new diseases. In Middle Europe, there is also a massive increase in
human infections caused by Hanta Virus, which is now spread also by insectivores (Soricidae)
in Austria.
‘A stitch in time saves nine’ and so it will be necessary to reduce the risks for infection and
to reduce the infection pressure by improving the habitats of wild animals. For preventing
diseases it also will be essential to remove infected animals or in some cases to reduce
appropriate wildlife stocks in a risk-free way. The consecutive monitoring of the wild animals’
health by examination of perished animals and by taking samples from unsuspicious animals
is the basis for early recognising and avoiding diseases in endangered wild animals species.
Finally, the following list of actions is proposed:

• Surveys, surveillance and monitoring of wild animal diseases.
• A stronger intersectoral collaboration between public health and veterinary institutions
al local, national and international levels.
• Adaptation of population size of wild animal species at suitable habitats.
• Information and training of hunters concerning important wild animal diseases.
• Disease awareness relating to ‘new infectious diseases’ (75% of these diseases are zoonoses!).
• Further investigations concerning ecology and biology of infectious agents, vectors and
intermediate hosts.
• Animal diseases are often media sensations. This circumstance often interferes with
competence and problem solving. Hence, appropriate ways of approaching the media
should be elaborated.
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Dynamics of infectious diseases according to
climate change: the Usutu virus epidemics in Vienna
Institute for Veterinary Public Health, Department for Farm Animals and Veterinary Public
Health, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria;
franz.rubel@vetmeduni.ac.at
Summary
As an example of the dynamics of infectious diseases in mid-latitudes, so far mainly observed

in the subtropics and tropics, we discuss the Usutu virus (USUV) epidemics in Vienna,
Austria. The USUV is an arbovirus, which is closely related to the West Nile virus. It caused
mass mortalities mainly of blackbirds (Turdus merula). Infections of mammalian hosts or
humans are rare. The USUV flavivirus persists in a natural transmission cycle between vectors
(mosquitoes) and host reservoirs (birds) and leads – once endemic in a population – to periodic
outbreaks. Following the recent work of Rubel et al. (2008) and Brugger and Rubel (2009),
we present an epidemic model to explain the USUV dynamics in Austria. Within the model
framework the USUV dynamics is mainly determined by an interaction of bird immunity
and environmental temperature. We demonstrate that the USUV model is able to simulate the
observations from the dead-bird surveillance 2001-2005. To investigate future scenarios, we
entered temperature predictions from five global climate models into the USUV model and
also considered four different climate-warming scenarios defined by the Intergovernmental
Panel on Climate Change, IPCC (20 different model-scenario combinations). Long-term
simulations cover the period 1901-2100. The results indicate that USUV will persist in the
host population after the epidemic peak observed in 2003, but the next major outbreak is
expected to occur not before 2019. USUV-specific annual blackbird-mortality time series
predict that the outbreak frequency increases successively from the beginning to the end of
the century. Additionally, we calculated the annually averaged basic reproduction number
for the period 1901-2100. The latter depict that undetected major outbreaks before 2000 were
unlikely, whereas it is likely that the USUV becomes endemic after 2040.
Keywords: arbovirus, basic reproduction number, simulation, epidemic model, global

warming
1. Introduction
The incidence and spread of previously tropical infectious diseases considerably increased
under the current situation of climate change (Harvell et al., 2002; Pfeffer and Dobler, 2009;
Gale et al., 2010) and global animal trade and migration (Pfeffer and Dobler, 2010). As
an example of how climate change may affect the emergence (or re-ermergence) in wild
birds of zoonoses that may have or gain public health relevance, the temperature dependent
dynamics of the Usutu virus (USUV), recently investigated by Rubel et al. (2008) and Brugger

and Rubel (2009), are presented here. USUV was detected the first time outside Africa in
2001 in Austria (Weissenböck et al., 2002). It is a member of the flaviviridae family of the
Japanese encephalitis virus complex and closely related to the more common West Nile
virus (WNV), dengue virus (DENV), Japanese encephalitis-virus (JEV) and yellow-fever
virus (YFV) (Weissenböck et al., 2002). USUV became well-known, because it caused mass
mortalities of blackbirds (Turdus merula) in and around the capital city of Austria, Vienna.
The established monitoring programme of the University of Veterinary Medicine Vienna
confirmed that USUV overwintered in Austria (Chvala et al., 2007). Although the initial
number of dead blackbirds was very low in 2001 and 2002, an epidemic peak was observed
during the extraordinary hot summer 2003 (Schönwiese et al., 2004). In the meantime,
USUV was also observed in other Central European countries such as Switzerland, Hungary
(Bakonyi et al., 2007), and Northern Italy (Lelli et al., 2008; Manarolla et al., 2010). Although
it was assumed that USUV does not cause severe or fatal disease in humans, in August 2009,
two human cases of Usutu virus neuroinvasive infection were documented in Italy (Pecorari
et al., 2009; Cavrini et al., 2009).
Arthropod-borne viruses (arboviruses), like USUV, persist in a natural transmission cycle

between vectors (here, mosquitoes of the Culex pipiens complex) and avian hosts. Once
endemic, it might cause periodic disease outbreaks. Rubel et al. (2008) developed a new
epidemic model for explaining the multi-seasonal dynamics of the USUV infection in Austria.
This model considers both the density-dependent seasonal dynamics of mosquitoes and birds
and the USUV bird-mosquito infection cycle. The dynamics of the mosquito population and
the virus reproduction rate depend strongly on the environmental temperature. Efficient
USUV transmission requires warm temperatures (which increase the reproduction and
biting rates of mosquitoes and decrease the extrinsic incubation period). To demonstrate
the relationship between warm temperatures and USUV, Figure 1 depicts the total number
of dead blackbirds associated with USUV infections (period 2001-2005) together with the
Hot days
0 3-5 7-8
0-1 5-6 8-10
1-3 6-7 > 10
Figure 1. Spatial distribution of the average number of hot days per year in Austria for the climate
reference period 1961-1990 (Auer et al., 2001) and blackbirds dead from USUV (white dots; Chvala et al.,
2007). Among some other species, about 140 blackbirds dead from USUV infections have been collected
between 2001 and 2005.

Dynamics of infectious diseases according to climate change
spatial distribution of the average number of hot days (days with temperature ≥30 °C) in
Austria. Conspicuously, the emergence of USUV infections is mainly related to those regions
around Vienna where the number of hot days exceeds 10 days/year. Most of the dead birds
were found in the most populated region in the district of Lower Austria, and only few in the
surrounding agricultural areas. A more detailed discussion on the dead-bird monitoring is
in Weissenböck et al. (2003) and Chvala et al. (2007).
Here we will respond to the following questions: is it possible to develop an epidemic model,
which can accurately simulate the observed USUV epidemics in Vienna? How might USUV
around Vienna develop according to global warming? Should we expect large-scale declines
in several bird populations as observed for the related WNV in North America (LaDeau et
al., 2007)? To answer the first question, we explain and discuss the USUV model developed
by Rubel et al. (2008), which calculates (once initialised with some USUV-positive mosquitoes
and exclusively forced by environmental temperature) time series of bird and mosquito
populations of various health states. It is shown that the epidemic model is able to reproduce
the observed numbers of dead birds very well. Therefore, the epidemic model will be forced
with long-term climate projections to answer the second and third question.
The applied climate dataset comprises temperature predictions from five different global
climate models (GCMs), to incorporate the uncertainty across climate models. However, each
GCM was based on the same initial conditions. We used a combination of these five climate
models with the four emission scenarios (defined by the Intergovernmental Panel on Climate
Change, IPCC) resulting in a total of 20 temperature predictions for the period 2006-2100 to
force our USUV model. The results of these multiple model runs are interpreted like results
from a stochastic model. Finally, we used temperature observations from a weather station in
Vienna to run the USUV model for the period 1901-2005. The major result from the epidemic
model runs is a time series of the basic reproduction number of the Usutu virus for the period
1901-2100. It depicts the possibility for a major outbreak as a function of environmental
temperature and herd immunity of birds.
2. The epidemic model
Our SEIR (susceptible, exposed, infectious, recovered) model simulates the seasonal lifecycles
of blackbirds (Turdus merula) and mosquitoes (Culex pipiens) and the inter-species USUV
infection cycle between birds and mosquitoes (Figure 2). The model has nine compartments
(i.e. health states): susceptible birds (SB), latent-infected or exposed birds (EB), infectious birds
(IB), recovered or immune birds (R B), dead birds (DB), mosquito larvae (LM), susceptible
mosquitoes (SM), latent-infected or exposed mosquitoes (EM) and infectious mosquitoes
(IM). The movements between health states are defined by 12 parameters, of which seven
depend exclusively on the environmental temperature. These are the birth and mortality
rates of larvae and (adult) mosquitoes, the infection rates between infectious mosquitoes and
susceptible birds and vice versa (cross-infection), and the reciprocal of the extrinsic incubation
period (here defined as infectivity rate of mosquitoes). Further parameters are the birth rate
of birds and the proportion of mosquitos’ non-diapausing, which are functions of the Julian
calendar day and the geographic latitude. Constant parameters are the mortality rate of birds,

Figure 2. Block diagram of the epidemic model applied to simulate the Usutu virus dynamics in Vienna.
The birth and mortality rates of mosquitoes (birth L , mortalityL , birthM, mortalityM), the infection
rates (infectionB, infectionM) and the infectivity rate of mosquitoes (infectivityM) are considered to be
temperature dependent.
the reciprocal of the intrinsic incubation period, and the infectivity rate of birds (i.e. the rate of
transition within birds from latently infected to infectious), the recovery rate of birds and the
death rate of birds. For more details on the mathematical parameter definition see Appendix 1.
Based on the same initial conditions we used a combination of these five climate models with
the four IPCC emission scenarios resulting in a total of 20 temperature predictions for the
period 2006-2100 to force our USUV model. The results of these multiple model runs are
interpreted like results from a stochastic model. Finally, we used temperature observations
from a weather station in Vienna to run the USUV model for the period 1901-2005. The major
results from the epidemic model runs are time series of the basic reproduction number of the
health states of birds and mosquitoes, respectively. They depict the possibility for past and
future outbreaks as a function of environmental temperature.
In doing so, the numbers of species in specific health state are calculated following the main
principle of building budgets. The future number of infectious mosquitoes, for example,
is calculated from actually available infectious mosquitoes minus the mosquitoes dying
within some specified time step (determined by the mortalityM rate) plus the newly infectious
mosquitoes (determined by the infectivityM rate). Thus, the dynamics of the 9 bird and
mosquito health states is mathematically formulated by 9 ordinary differential equations (not
shown), which are solved numerically using daily time steps (R source code see Appendix 2).

3. Parameter estimation
The extent to which an epidemic model simulates reality depends strongly on the assumed
parameters (birth, mortality and transmission rates). Note that the model parameters are
generally defined per capita and per day and will be described in detail in the following
sub-sections.
3.1 Bird parameters
Because more than 90% of all birds, which died from USUV infections, were blackbirds
(Turdus merula), we focus on this species. Blackbirds are widespread in woodland, but also
one of the most striking birds in urban gardens. Their average life expectancy is about 2
years, but may exceed 20 years in exceptional cases. Knowing the lifetime of blackbirds, the
mortality rate can be estimated as its reciprocal value. Hatchwell et al. (1996) specified annual
mortality rates of 0.34 per year and 0.52 per year in woodland and farmland, respectively.
For the land cover of the area around Vienna we applied a mean mortality rate of 0.43 per
year corresponding to mortalityB = 0.0012 days−1.
In Central Europe blackbirds deposit eggs two to four times per year. Schnack (1991)
investigated the breeding success and clutch sizes of blackbirds in 17 city parks in Vienna
and in an adjacent lowland forest. On average a clutch size of 4.1 eggs was estimated for the
city of Vienna and 4.6 eggs for the woodland. The breeding success (the number of fledged
nestlings per eggs laid) was 22.4% for urban blackbirds and 30.7% for forest blackbirds. Similar
results were documented by Tomialojc (1993 and 1994) for blackbirds in Poland (mean clutch
size of 4.5 eggs, emerging nest losses of 50-92% with mean 68%). On averaged 2.5 young per
pair fledged yearly, whereas the most successful pairs reared 8-9 young (Tomialojc, 1994).
We assume 2.5 young per pair as bird birth rate. This yields a per capita birth rate of 1.25 per
year or birthB = 0.00342 days−1.
Although the mortality rate is assumed to be uniformly distributed over the year, a seasonal
cycle was fitted to the observed birth rate. Figure 3 (left) depicts the observed frequency
distribution of the blackbird nestlings (bars), as compiled from counts of blackbird clutches in
Poland (Tomialojc, 1994), and the fitted theoretical distribution (line). Originally, Tomialojc
(1994) published absolute numbers of clutches for two observational periods, which were
averaged and shifted by 10 days to account for breeding. As depicted in Figure 3 (left), the
distribution is skewed to the right. Therefore, a gamma distribution was selected to describe
the observations. By multiplying it with the average annual birth rate, we derived the birth
rate as a function of the calendar day as shown in Figure 3 (right).
The breeding density in the city of Vienna varies in the range of 10-210 pairs/km2 (Schnack,
1991), corresponding to a bird density of 20-420 birds/km2. Two typical black bird habitats
outside the city of Vienna were investigated by Wichmann and Zuna-Kratky (1997). The first
habitat is an area covered with vineyards and fallow lands, and the bird density was 77-122
birds/km2. The second habitat, a mixed forest with embedded grasslands, showed a slightly
higher bird density of 104-155 birds/km2. These densities are much higher than the large-scale
blackbird density for central Europe, for example, of 25 birds/km2 as estimated for the entire

Figure 3. Observed relative frequency of blackbird nestlings (Tomialojc, 1994) with fitted gamma
distribution (left) and bird birth rate as function of the Julian calendar day (right).
region of Germany (Schwarz and Flade, 1989). Considering both the large-scale density and
the local densities of favoured blackbird habitats, we assumed an average blackbird density
for the area of USUV emergence of about 50 birds/km2. We used this value to define the
carrying capacity of birds.
An accurate estimation of the bird mortality due to USUV infections is difficult. From the
data of the dead-bird surveillance we estimated that about 30% of the infected blackbirds
died (Weissenböck et al., 2002).
3.2 Temperature-dependent transmission parameters
According to the cross-infection process depicted in Figure 2, two transmission parameters,

the forces of infection, must be determined. Here, the forces of infection are abbreviated by
the terms infectionB and infectionM, which are functions of the mosquito biting rate k and the
probabilities pB and pM that the USUV is transmitted by a bite. As for WNV, Culex mosquitoes
are mainly responsible for USUV transmission. On average these mosquitoes are biting every
1-5 days, corresponding to biting rates of k = 0.2-1 days−1; see for example the overview
given by Cruz-Pacheco et al. (2005). Temporal changes in the effectiveness of transmission
essentially delineate the seasonality of WNV and USUV activity and are triggered by the
environmental temperature. For a description of this process, the temperature dependence
of the duration of the gonotrophic cycle (i.e. the development cycle of mosquitoes comprising
blood meal as well as development and deposition of eggs) as presented by Reisen et al. (2006)
was used. The function describing the biting (i.e. contact) rate was fitted to the reciprocal of the
duration of the mosquito gonotrophic cycle (Rubel et al., 2008). The functional relationship is
similar to Figure 4A, because the birth rate of the larvae (birthL) is assumed to be a multiple
of the biting rate k. The biting rate increases with increasing temperature according to a
logistic function. The infection rates are proportional to the product of the biting rate and the
transmission probabilities. Typical WNV probabilities as proposed by various authors vary
of 0.02-0.24 (virus transmission by infectious birds to susceptible mosquitoes) and 0.8-1.0
(virus transmission by infectious mosquitoes). We determined the unknown transmission
probabilities of USUV by fitting the model to observations, resulting in a transmission

Figure 4. Observed mosquito birth and mortality rates as function of temperature. A. Function birthL
fitted to data from Reisen et al. (2006) and adjusted after Rubel et al. (2008). B. Function mortalityL fitted
to data from Bailey and Gieke (1968). C. Function birth M fitted to data from Reisen (1995). D. Function
mortalityM fitted to data from Reisen (1995). Note, that the parameter functions have been selected to
yield parameters for larvae that are exact one order of magnitude higher than for adult mosquitoes.
probability by infectious birds of pB = 0.125 and by infectious mosquitoes of pM = 1.0 (both

are within the range given for WNV).
3.3 Temperature-dependent mosquito parameters
The temperature dependence of mosquito birth and mortality rates (of both larvae and adult
mosquitoes) was investigated mainly in the 1960s and 1970s. Unfortunately, most of these
studies do not provide functions as required for process models. Therefore, one of our goals
was to find general functions describing the relationship between mosquito population

parameters and environmental temperature. A selection of functions fitted to data sets

published by various authors is depicted in Figure 4.
Figure 4A shows the birth rate of larvae birthL , a synonym for the egg-deposition rate, which
is modelled by the scaled reciprocal of the gonotrophic cycle after Reisen et al. (2006). We
selected the scaling factor so that the average birth rate birthL = 0.537 days−1, as proposed by
Wonham et al. (2004), is reached at T = 25 °C. Typical mortality rates of larvae are shown
in Figure 4B, where a function was fitted to data from Bailey and Gieke (1968). Alternative
functions (not shown) were used for example by Eisenberg et al. (1995) or Shaman et al. (2006).
Most studies are available for the temperature dependent birth rate of adult mosquitoes birthM
(that is, the development rate of immatures). These studies comprise laboratory experiments
for Culex quinquefasciatus and Aedes aegypti (Rueda et al., 1990), a regression line for Culex
annulirostis (Rae, 1990) as well as discrete values for Culex tarsalis (Reisen, 1995) and for Culex
pipiens molestus (Olejnícek and Gelbic, 2000). Logistic functions birthM were fitted to all of
these data sets and subsequently were evaluated in sensitivity studies (results not shown). As
an example, the function fitted to the data published by Reisen (1995) is depicted in Figure
4C. Finally, Figure 4D shows the mortality rate mortalityM, again as fitted to observations
from Reisen (1995).
From inspection of Figure 4, it became clear that that the functions for the population
parameters of the mosquito larvae are of similar shape, but about one order of magnitude
higher than those for the adult mosquitoes. Using this allowed us to generalise the mosquito
birth rates by using the same logistic (S-shaped) function for both, larvae and adult mosquitoes.
Again, only one U-shaped function was selected to describe both mortality rates (Figures 4B
and 4D), which differ by a factor of ten.
3.4 Temperature-dependent extrinsic-incubation period
The extrinsic-incubation period (i.e. the time from an infectious blood meal until the mosquito
can transmit an acquired arbovirus infection) is an important parameter determining the
vector capacity. It is the reciprocal of the rate of virus replication within an infected mosquito
vector, here called infectivity rate. The parameter infectivityM is highly temperature-dependent
(Cornel et al., 1993; Dohm et al., 2002; Turell et al., 2002; Reisen et al., 2006).
Because the virus-replication rate for USUV is unknown, we relied on the results from WNV.
We used data from Reisen et al. (2006) to fit the functions depicted in Figure 5. Thus, the
extrinsic-incubation period decreases with increasing temperature. Long periods of warm
temperatures amplify flavivirus transmission. Vice versa, low temperatures can reduce the
flavivirus transmission or even interrupt it when the extrinsic-incubation period exceeds
the mosquito lifetime.
3.5 Diapause (hibernation of mosquitoes)
Only non-diapausing mosquitoes contribute to the reproduction and the USUV transmission
cycle. Therefore, it is of fundamental importance to know the fraction of non-diapausing
mosquitoes. Quantitative investigations on the diapause of Culex mosquitoes are rare because

Figure 5. Observed infectivity rates (dots) and fitted function for infectivityM (left) and its reciprocal, the
extrinsic-incubation period (right). Both are functions of the environmental temperature and fitted to
data by Reisen et al. (2006).
they had not played an important role as vector until the 1999 WNV outbreak in New York
(USA). One study was published by Eldridge (1968), who depicted the proportion of non-
diapausing mosquitoes as a function of photoperiod (daytime length) and temperature
(Vinogradova, 2000). A second paper was presented by Spielman (2001), who re-analysed
50-year-old-mosquito data from Boston (300 km north of the WNV outbreak in New York)
to deduce a relationship between diapause and photoperiod (dots in Figure 6, left).
Figure 6. Observed fraction of diapausing Culex pipiens mosquitoes as function of the daytime length
after Spielman (2001) with fitted function (left) and annual cycle of the daytime length in hours for the
geographical latitude of Vienna, Austria (right). At daytime lengths above 14 hours and 10 minutes (May
to August) more than 75% of the mosquitoes are active.

We fitted functions to both data sets: A 2-dimensional function (not shown) to the data of
Eldridge (1968) and a 1-dimensional function to the data of Spielman (2001). Simulations
demonstrated that their application yielded similar model results. This is not astonishing
because of the natural correlation between the annual temperature cycle and the photoperiod.
We applied the simpler relationship after Spielman (2001). The logistic function for the
description of the fraction of active mosquitoes, i.e. the non-diapausing mosquitoes, is
depicted in Figure 6 (left). It depends on the daytime length in hours, which is calculated
from the astronomic declination (a function of the calendar day) and the geographic latitude.
The annual cycle of the daytime lengths in Vienna is depicted in Figure 6 (right).
4. Climate forcing of the observational period 2001-2005
As discussed above, the epidemic model is forced by temperature data, which determine
the transmission rates infectionB and infectionM via the mosquito biting rate k as well as the
mosquito parameters birthL , mortalityL , birthM, mortalityM and infectivityM.
The air-temperature measurements from the automatic weather station located at the
University of Veterinary Medicine, Vienna, were used. These measurements are available
for the period 1997 to present and are representative for the study area around Vienna. The
temporal resolution of the measurements is 10 minutes, providing the opportunity to run
the epidemic model with time steps corresponding to this resolution.
On the other hand, the numbers of dead birds collected during the USUV monitoring
program (Chvala et al., 2007) are only representative on a weekly or (better) monthly time
scale. Our goal was to reproduce and explain the observed multi-seasonal dynamics of
USUV infections. Therefore, to account for the resolution of the observations, the model was
forced by monthly averaged temperature data. Nevertheless, to assure numerical stability,
the model must run with time steps of 1 day or less. We used spline functions to interpolate
the monthly data to the model time step. In fact, smoothed data were used to force the
model. Figure 7 (upper panel) depicts the time series of the temperature observations in
Vienna for the study period 2001-2005, whereas the grey line represents the daily and the
dark line the monthly averaged (smoothed) temperature. Figure 7 (lower panel) points out
the temperature anomaly: the deviation from the 10-year mean 1997-2006. Conspicuously
positive temperature deviations during the mosquito-activity period were observed for May
to July 2002 and for May to September 2003, whereas no longer periods of extraordinary high
temperatures were observed during the other years. Subject to the temperature dependence of
mosquito parameters discussed in the previous section, high USUV activities were expected
for 2002 and 2003. Most interesting is the year 2003, where the June to August temperatures
exceed the long-term mean by more than 3 °C.
5. Short-term simulation results for the period 2001-2005
The first results of the model simulations were time series of health states of birds and
mosquitoes. Figure 8 (upper panel) depicts the normalised numbers of mosquito larvae and

Figure 7. Observed daily and monthly averaged temperatures (upper panel) and monthly temperature
anomalies (lower panel) in ° C. Location Vienna, period 2001-2005. Note the positive anomalies in spring
and summer 2003.
Figure 8. Simulated time series of mosquitoes for the period 2001-2005 (proportion of mosquitoes/bird).
Upper panel: larvae LM (dotted line) and susceptible mosquitoes SM (solid line) for carrying capacity K M
= 100 mosquito larvae/bird. Lower panel: infectious mosquitoes IM.
adult mosquitoes for the investigation period 2001-2005. While the mosquito population
during the years 2001, 2004 and 2005 seemed typical, it was 2-3 times higher in 2002 and 2003.
The latter were caused by the long warm periods in these years (Figure 7). The normalised

number of infectious mosquitoes, IM (Figure 8, lower panel) was influenced by the extrinsic-
incubation period, which decreases with increasing temperature (Figure 5).
Figure 9 (upper panel) depicts the proportion of birds in the health states susceptible SB,
immune R B, as well as the total population NB. A strong increase of the immune birds R B =
0.7 (70%) was simulated for 2003 in correspondence to the epidemic peak in the same year.
Subsequently, R B decreases to 45% until the end of 2004 and to 30% until the end of 2005.
Analogous to the illustration of the time series of mosquitoes, Figure 9 (lower panel) depicts
the proportion of infectious birds with the same epidemic peak in 2003.
For a verification of simulations with observations (Figure 10), the normalised model results
were scaled to fit the observations. This has been done by multiplying the model output, i.e. the
proportion of birds in the various health states (Figure 9), by a factor of 385 birds. This factor is
the carrying capacity of birds KB,obs = 385, which is called observed, because it follows from the
number of observed dead blackbirds. After this scaling, our model described the observations
quite well. Note that the dead-bird monitoring was organised only for the summer months
June to August, while the model simulates continuously in time. Feasible comparisons may
therefore only be done for the three summer months. Nevertheless, the model simulations
indicate that blackbirds die from USUV during the whole mosquito activity period May to
October. Based on the estimated blackbird density of 50 birds/km2 (see Section 3.1) and an
investigation area around Vienna of about 3,500 km2, the true carrying capacity of blackbirds
is about K B,true = 175,000 birds. From the fraction KB,obs/KB,true it follows that only 0.2% of
the USUV positive birds were detected by the dead-bird monitoring program.
Figure 9. Simulated time series of blackbirds for the period 2001-2005 (proportion of birds related to K B).
Upper panel: susceptible birds SB (dotted line), immune birds R B (bold solid line) and total birds NB (thin
solid line) for KB = 1. Lower panel: infectious birds IB.

Figure 10. Time series of monthly averaged dead blackbirds for the period 2001-2005. Upper panel:
observed; lower panel: simulated by the epidemic model (scaled with K B,obs = 385). Note that the number
of dead birds modelled for 2005 is below 1, but non-zero.
6. Climate forcing for the extended period 1901-2100
To simulate future USUV outbreaks, we selected the Tyndall Centre for Climate Change
Research dataset, TYN SC 2.0 (Mitchell and Jones, 2005). It is based on 4 scenarios defined by
IPCC and described in the Special Report on Emission Scenarios (SRES). This dataset provides
the 5 climate parameters temperature, diurnal temperature range, precipitation, water-vapour
pressure, and cloud cover; but we only used the temperature data. Information is presented
on a 0.5° grid, equidistant in geographical latitude and longitude that covers the world land
surface and is available with a monthly time-step for the period 2001-2100. Thus, the TYN SC
2.0 dataset comprises a total of 20 climate-change model runs, combining 4 possible future
worlds using SRES scenarios with 5 state-of-the-art climate models (Mitchell et al., 2004).
The emission scenarios (Figure 11) were developed in the mid 1990s and are based on 4
different narrative storylines to describe consistently the relationships between the forces
driving emissions and their evolution and to add context for the scenario quantification
(IPCC, 2000). The four SRES storylines represent different world futures in two dimensions:
a focus on economic or environmental concerns, and global or regional development. The
variables in each model include population growth, economic development, energy use,
efficiency of energy use, and mix of energy technologies, respectively. The TYN SC 2.0 dataset
considers the scenarios A1 (exactly A1FI), A2, B1 and B2. On the other hand, five general
circulation models were used to simulate climatic changes: the Hadley Centre Coupled
Model Version 3 (HadCM3), the National Center for Atmospheric Research-Parallel Climate
Model (NCAR-PCM), the Second Generation Coupled Global Climate Model (CGCM2), the
Industrial Research Organization – Climate Model Version 2 (CSIRO2) and the European
Centre Model Hamburg Version 4 (ECHam4).

Figure 11. Brief characterisation of the main Special Report on Emission Scenarios (SRES) storylines after
IPCC (2000) and Arnell et al. (2004).
We used only time series of monthly temperatures of the grid-point 392/277 of the TYN SC
2.0 dataset (the so-called grid-point Vienna), which is representative for the geographical
region between 16.0-16.5° longitude and 48.0-48.5° latitude. Figure 12 depicts time series
of the average annual temperature for the grid-point Vienna, grouped by scenarios (upper
panel) and climate models (lower panel). All predicted temperatures show a clear trend toward
warming. The average predicted increase of the temperature during the 100-year period,
calculated from the linear trend, ranged from 2.6-6.1 °C (across emission scenarios) and
from 2.3-6.8 °C (across climate models). For the 20 model-scenario combinations, the range
in average annual predicted warming across the century was 0.8 °C (PCM model with B1
scenario) to 11.6 °C (ECHam4 model with A1 scenario). The lowest temperature increase of
2.3 °C (0.8-3.8 °C) is simulated by the B1 scenario. It represents an environmentally oriented
world with emphasis on global solutions to economic, social and environmental sustainability.
Two specific features of climate model predictions have to be considered when using it to
study the evolution of biological systems (e.g. epidemic outbreaks). Firstly, climate predictions
for the next 100 years merely simulate a possible temperature based on a scenario which is
also a prediction. Uncertainties of these climate predictions are made evident by the range
of prediction, depending on the scenario and model used. Secondly, climate models are
representative for a specific spatial resolution, here of 0.5° equidistant in geographical latitude
and longitude. Thus, the temperature at grid point Vienna is representative for an area of about

Figure 12. Time series of average annual temperature predictions for different emission scenarios (upper
panel) and climate models (lower panel). Numbers in brackets denote the temperature range predicted
by different climate models (upper panel) and different emission scenarios (lower panel). Grid-point
Vienna, period 2001-2100.
70×50 km2 with a mean height above sea level of 261 m. The grid-point Vienna comprises,
for example, the foothills of the Alps, the Vienna Forest, and planes like the Tullner Feld (i.e.
the resolution is too coarse for our study area). USUV cases have been observed only in the
flat country where temperatures are high. These temperatures were measured by our weather
station located at 153 m above sea level. Therefore, the temperatures simulated by climate
models were adjusted to the measured temperatures (downscaled) to describe this small-
scale region (Rubel et al., 2008). Note that it is essential to use the (generally slightly higher)
downscaled temperature predictions to force the USUV model. Otherwise the effect of the
climate warming signal would be compensated by the underestimation of the temperatures
predicted by the large-scale climate models.

7. Long-term simulation results for the period 1901-2100
The time series of predicted annually averaged numbers of birds dead because of USUV is
depicted in Figure 13. The upper panel shows the mean proportion of dead birds together
with the 95% confidence interval (grey areas) calculated from 5 model runs forced by the
GCM predictions for the worst-case emission scenario A1. The lower panel shows the same
for the best-case scenario B1.
The main result is that USUV was predicted to survive in almost all model simulations except
those simulations where the USUV model is driven by temperatures from PCM using the A1
and A2 scenarios. This virus extinction leads to the higher variation of the proportion of dead
birds in the A1 scenario. After the epidemic peak 2003, only minor outbreaks are simulated
until the next major outbreak in 2019. The amplitude of the outbreaks, here the annually
accumulated proportions of dead blackbirds, vary accordingly to the predicted temperatures.
The higher temperature increase of the A1 scenario results in the higher predicted annual
USUV specific mortality risk in the blackbirds. Moreover, forcing the USUV model with
Figure 13. Average (central line) and 95% confidence interval (grey areas) of USUV-specific mortality of
blackbirds around Vienna. Emission scenario A1 (upper panel) and B1 (lower panel).

temperatures from two climate models (ECHam4 and HadCM3 with A1 scenario) result in an
endemic situation with a constant level of about 11% dead birds in the year 2050. Typically, a
cyclic annual mortality pattern was predicted through the years – but the predicted amplitude
of the cycle of annual mortality decreases later in the century especially of the A1 emission
scenario. For the A1 scenario on average 60% of all birds acquire immunity until the end of
the century. But also for the best-case scenario B1, a proportion of 50% of the birds acquire
lifelong immunity.
Additionally, we depict the USUV dynamics by the annual basic reproduction number R0
as derived and discussed by Rubel et al. (2008). Figure 14 shows the time series of R0 for
the period 1901-2100. While IPCC scenarios have been used to predict the future, observed
temperature values were taken to calculate a projection to the past. Major outbreaks may only
occur for R0 >1, firstly calculated for the observed USUV epidemics 2001-2005. Thus, our
backward projection using observed temperatures shows that USUV was not able to cause
major outbreaks in the past. Nevertheless, minor outbreaks below some detection threshold
were possible, because of the annual cycle of R0 (not shown).
Figure 14. Mean annual basic reproduction numbers for the IPCC worst-case scenarios A1 (upper panel)
and best-case scenarios B1 (lower panel). Linear trends for the period 1901-2100 indicate an increase of
R0 = 0.61 per 100 years for the A1 and R0 = 0.52 per 100 years for the B1 scenario.

8. Conclusions
As an example for viral zoonoses affecting wildlife and domestic animals, the dynamics of
the well-documented USUV epidemics in Vienna was investigated. After the epidemic peak,
more than 70% of the bird population acquired immunity. However, the immunity decreases
very quickly and already some years after the epidemic peak in 2003 a new major outbreak can
occur, depending only on the environmental temperature. Other environmental parameters
such as precipitation or flooding seem to play only a minor role (because our model fitted
well without them). The 100-year flood in Vienna in August 2002, for example, had no effect
on the USUV epidemics. Similar observations were made in the U.S. in connection with the
hurricane Katrina and WNV (Farnon, 2006). The so-called floodwater mosquitoes are minor
effective vectors for WNV and USUV.
From 20 scenario-model combinations we examined, only two were predicted to lead to

extinction of USUV (PCM, A1 and A2 scenarios). The other 18 combinations lead to endemicity
in Central Europe through the end of the century. It is therefore very likely, that the virus
survives in Central Europe until the end of the century. We do not further investigate the
possibilities for local extinction because we assume that the USUV is continuously introduced
by migratory birds. Thus, the most important epidemiological quantity determining major
outbreaks is the basic reproduction number R0. It predicts optimal environmental conditions
for USUV outbreaks due to global warming in about 10 years. On the other hand, undetected
USUV outbreaks before the observed outbreak in 2003 are very unlikely.
Our model explains the USUV transmission by combining both the effect of the immunity
status of the bird population and effects of the environmental temperature. Additionally,
the parameter estimation was confirmed by the results of a Bayes analysis (Reiczigel et al.,
2010). Thus, we propose the application of the model presented here for studies on WNV
transmission in North America.
Our estimations seem realistic when compared to the observed large-scale declines in the
bird populations caused by WNV infections in North America (LaDeau et al., 2007). Possible
effects of global warming concerning bird and mosquito populations due to changes in land
cover, availability of food or other environmental factors have not been considered; i.e., the
carrying capacity of birds K B and mosquitoes K M were assumed to be constant during the
investigation period 2001-2100. Further, we did not consider either vertical transmission of
USUV in the mosquito vectors or horizontal transmission in susceptible birds. This decision
was made because there exist no data at all on these particular transmission scenarios in
USUV infections. In addition, these modes of transmission have been experimentally shown
in infections with the related WNV, but their relevance in field infections seems negligible
(Baqar et al., 1993; Komar et al., 2003).

A comprehensive verification of the arbovirus model might be realised in a few years when
longer time series of observations are available. Especially, the population dynamics of
mosquitoes of the Culex pipiens complex have to be verified. Unfortunately, there are no
current entomological field studies in Austria. Also, the assumption that the extrinsic
incubation period of USUV (which has never been investigated in laboratory experiments)
is similar to that of the closely related WNV has to be verified. Other recently published
verification datasets comprise the abundance of blackbirds. Steiner and Holzer (2008), for
example, investigated the decline of blackbirds at 6 locations in Vienna. They found big local
differences in time and extent of mass mortality in blackbirds. While the decline in bird
populations was about 58% in some regions near the Danube River, in other regions more
than 94% of blackbirds disappeared. This disagrees with the experience of G. Loupal (personal
communications), who stated that the blackbird population was not dramatically influenced
by the USUV epidemics, as it has also been confirmed by observations.
Acknowledgements
The authors are very grateful to Prof. Dr. Herbert Weissenböck and Dr. Sonja Chvala-
Mannsberger, both University of Veterinary Medicine Vienna, for their dead-bird surveillance
data and to Dr. Ingeborg Auer from the Austrian weather service (Zentralanstalt für
Meteorologie und Geodynamik, ZAMG) for kindly providing the map of hot days as well
as the time series of observed temperature data. The Tyndall Centre for Climatic Change
Research at the University of East Anglia, UK, provided the GCM projections.
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Appendix A The USUV model
Model equations
Health states

Main parameters
Additional parameters

Appendix B R-Source-code for the USUV model

Other contributions

The utility of GIS in studying the distribution of
Bovine Tuberculosis in wild boar (Sus scrofa) and red
deer (Cervus elaphus) in Central Portugal
João R. Alberto1, José M. Aranha2, João P. Serejo3, Alice Amado4 and Madalena
Vieira-Pinto1
1Departamento de Ciências Veterinárias, Lab. Inspecção Sanitária, CECAV-UTAD,
Universidade de Trás-os-Montes e Alto Douro, Apartado 1013, 5001-801 Vila Real, Portugal;
joao_c_alberto@hotmail.com
2Departamento de Ciências Florestais, Lab SIG, Universidade de Trás-os-Montes e Alto
Douro, Apartado 1013, 5001-801 Vila Real, Portugal

3Câmara Municipal de Idanha-a-Nova, Gabinete Médico Veterinário, 6060-163 Idanha-a-
Nova, Portugal
4Laboratório Nacional de investigação Veterinária (LNIV), Estrada de Benfica, 701, 1549-011
Lisboa, Portugal
Summary
Tuberculosis in game animals has gained, in the past few years, an increasing significance in
the central region of Portugal, mainly in wild ungulates such as red deer (Cervus elaphus) and
wild boar (Sus scrofa). Geographical Information Systems (GIS) technology is becoming an
essential component of modern disease surveillance systems. In this study GIS was used to
evaluate the geographical distribution of Bovine Tuberculosis (BT) in large game species in
Central Portugal. Sampling plots for GIS were mapped by means of a GPS (Global Positioning
System) receiver in Idanha-a-Nova county (lat 39° 55’ N: long 7° 14’ W) from November
2008 to February 2009. Over this period, 526 animals (337 red deer (Cervus elaphus), 142
wild boars (Sus scrofa), 29 fallow deer (Dama dama), and 18 mouflon (Ovis musimon)) were
analysed for BT lesions, during meat inspection. From these harvested animals, 73 (13.88%)
carried BT compatible lesions, which were later confirmed by laboratorial analysis. Data
collected during fieldwork were assigned to each sampling plot location, in order to enable
geostatistical analysis. The analyses of the BT intensity map, created by GIS, for wild boar
and red deer hunted in Central Portugal, allow the conclusion that the main BT-afected
areas were located at the south-east area of the county. These areas should be the first ones
under veterinarian scrutiny with a view to control or reduce disease spread and prevalence.
GIS provided an important tool to define objective strategies for preventing the spread of
infectious disease.
Keywords: tuberculosis, wild ungulates, Mycobacterium bovis, zoonoses, GIS

João R. Alberto, José M. Aranha, João P. Serejo, Alice Amado and Madalena Vieira-Pinto
1. Introduction
Zoonoses are a matter of concern to the public health and to the economy, and the role of
wildlife in the epidemiology of these diseases is an issue of increasing interest and importance
and its surveillance among wildlife is a research and public health challenge (Childs et
al., 2007). According to Kruse et al. (2004), zoonoses with a wildlife reservoir represent
a major public health problem, affecting all continents. Also, the possibility of persistent
cycling of infection in wildlife reservoirs could compromise the success of domestic animals
disease control programmes, with additional concern to veterinary authorities, wildlife
conservationists and managers (Bengis et al., 2002; Gortázar et al., 2007). Classical Swine
Fever and Brucellosis in wild boars and Bovine Tuberculosis (BT) persistence in badgers are
well-known examples (Bengis et al., 2002; Meldrum, 2003). With respect to BT, it is known
that this disease is becoming an increasingly important throughout Europe in major game
animals, like wild boars and red deer. On the Iberian Peninsula, according to Hermoso de
Mendoza et al. (2006), wild stock (e.g. wild boar and red deer) shows evidences of sharing
tuberculosis with domestic cattle, thus endangering human health and leading to economic
losses. As already reported by Santos (2006) and Duarte et al. (2008), BT in major game
(mainly in red deer and wild boar) is often found in central Portugal. This requires a rapid
intervention by the veterinary competent authority in order to reduce the prevalence. Cost
effective prevention and control of BT requires an interdisciplinary and holistic approach
and international cooperation, where surveillance, laboratory capability, research, training
and education, and communication are key elements (Kruse et al., 2004).
Geographical Information Systems (GIS) enable the incorporation of different spatial data like
geographical, farm locations and diseases distribution due to its capability to record information
from different sources and its potential for data management and manipulation (Antenucci et
al., 1989; Goodchild et al., 1992). GIS also facilitates the epidemiological relationship analysis
among these variables, which is of major importance to the epidemiological investigation
of wildlife diseases (Clark et al., 1996; Kistemann et al., 2002; Schröder, 2006). In addition,
output data generated by GIS in map format has the particular advantage of allowing implicit
representation of spatial dependence relationships (Clark et al., 1996; Schröder, 2006; Xavier
et al., 2007) in an intuitive manner. Advances in technology and decreasing associated costs
has lead to an increasing use of GPS (Global Positioning System) as a tool for mapping wild
animals moving across natural areas or for geo-referenced data collection (Schlecht et al.,
2004; Gorman et al., 2008). Thus, GPS has been used for mapping large game hunting activity
and as a GIS data source.
Although some studies indicate that GIS is an important tool in wild stock management
(e.g. Radeloff et al., 1999), no references were found about the GPS-GIS use in animal disease
analysis and control. Hence, GIS and GPS technologies were used in this study so as to
evaluate the geographical distribution of Bovine Tuberculosis in large game species in Central
Portugal.

The utility of GIS in studying the distribution of Bovine Tuberculosis in Central Portugal
2.1 Sampling area
The study was performed in Idanha-a-Nova county placed in the central-east part of Portugal
(lat 39° 55’N: long 7° 14’W) with 1,412.7 km2 and 10,561 inhabitants. Located in a plateau
region, it has a large border line with Spain defined by the river Erges on the east and the
river Tagus in the south.
This is a typical Portuguese rural area with approximately 50% of land dedicated to agriculture
(43% dry agriculture, 9% watered agriculture, and 5% graze land), 30% representing forested
areas (mainly oaks) and 13% shrub land and sparse vegetation.
Idanha-a-Nova is considered to be one of the best hunting areas in Portugal with numerous
game estates, many of which also breed cattle, sheep in an extensive free ranging system,
grazing in pastures which are also used by wild artiodactyls.
2.2 GIS methodology
Using information produced and presented by Portuguese cartographic authorities (National

Geographic Institute: www.igp.pt; Military Geographic Institute: www.igeoe.pt) such as
topographic plans, contours and CORINE Land Cover, a GIS (ArcGis 9.x, Arcinfo version)
was created.
Sampling plots were mapped by means of a GPS (Global Positioning System) receiver in
Idanha-a-Nova county from November 2008 to February 2009, in order to geo-reference all
large game events, suitable to be used for updating the GIS. Within this period, a total of 526
animals (337 red deer (Cervus elaphus), 142 wild boars (Sus scrofa), 29 fallow deer (Dama
dama), and 18 mouflon (Ovis musimon)) hunted in 20 organised drive hunts, were analysed
during post mortem meat inspection for Bovine Tuberculosis compatible lesions. Compatible
BT lesions were defined as described by Martín-Hernando et al. (2007).
Samples with BT compatible lesions were collected and sent to the Laboratório Nacional de
Investigação Veterinária (LNIV: National Veterinary Research Laboratory) for confirmation
by means of histopathology and other tests like PCR-REA system (Niemann et al., 2000).
Using GPS collected positions, a vector file (point shape file) was created with all sampling
plots locations. Data collected during fieldwork (e.g. tuberculosis compatible lesions) were
assigned to each sampling plot location, in order to enable descriptive statistics calculation and
geostatistical analysis (Schröder, 2006). Using the percentage of tuberculosis compatible lesions
calculated for each sampling plot and geostatistical analysis (performed using Geostatistical
Analyst 2.0 for ArcGIS 9.x. ArcInfo version) a continuous map related to disease spread was
created, enabling extending the results to the entire study area thus creating continuous
disease intensity maps represented by a colour intensity gradation (Schröder, 2006).

From the total of 526 animals hunted and examined during post mortem meat inspection,
73 (13.88%) presented BT compatible lesions: 45 red deer and 28 wild boar. Figure 1 and 2
represent two examples of BT compatible lesions observed during this study and located in
mesenteric lymph nodes and lungs, respectively.
Figure 1. BT compatible lesions (white arrow) located in mesenteric lymph node of one red deer.
Figure 2. BT compatible lesion located in lung of one red deer.

GIS analyses of the field work allowed the creation of a continuous disease intensity maps
represented by a greyscale intensity degree scale as presented in Figure 3. The percentage
values represent the affected BT animals in the total of hunted animals per drive hunting day.
Figure 3 shows, that tuberculosis in wild boars (Figure 3, left side) is scattered throughout the
county (circle dimension). After geostatistical calculation (grey gradient), it is possible to see
a major vector from the south-south-east in a northward direction, indicating an important
level of tuberculosis spread. Tuberculosis in red deer (Figure 3, right side) is confined to the
south-south-east area. Both maps indicate that the main areas affected with tuberculosis in
wild boar and red deer are located in the south-east area of the county. These areas should
be the first to be put under veterinary surveillance and for which strategies to reduce disease
spread and prevalence should be developed.
According to Curtis (1999), one of the main advantages of using GIS in a disease surveillance
programme is that this technique can be used as a first step for identifying areas that need
improvement in their surveillance scheme. Also, the simple use of GIS as a map of known
cases allows identifying the location of those environments considered relevant to the disease
or health risk. This allows mental/intellectual exploration of the whole ecological situation,
but especially in space and time, which are the most important dimensions of the diseases of
wildlife (Pfeiffer and Hugh-Jones, 2002).
Our study shows that the south-south-east is home to much higher densities over a larger
and more continuous area than is the case for the north regions. Also, more hunted animals
are processed in the same area and the offal and by-products are often thrown out to feed
the vultures in places equally accessible to other species like wild boar. We believe that this
irresponsible act is responsible for perpetuation of this situation.
A National Eradication Programme for bovine tuberculosis has been in operation since 1987
and is presently based on systematic tuberculin testing and slaughter with bacteriological
Figure 3. Bovine tuberculosis intensity map for wild boar and red deer hunted in Central Portugal.

diagnosis carried out at the National Reference Laboratory (NRL-LNIV) and permanent
abattoir surveillance (Duarte et al., 2008). However, this programme can be undermined by
game species if they act as reservoirs and spreaders of the agent. It is very important to make
a GPS-GIS surveillance and control scheme and link it with the location of cattle farms so
as to optimise resources in the right places and at the right time. A country scale reporting
system, assessing an acceptable level of health in game species is recommended in a EU
directive (92/45/CEE) (Artois, 2003).
4. Conclusions
Investigation of diseases in wild animals presents a special challenge to veterinary

epidemiologists as the locations of individual animals within the populations at risk are
much less predictable than are domestic animals. GIS can be considered an important tool
to define objective strategies to prevent the spread of the infectious disease.
The following areas in which GIS and special GIS-functions could be incorporated include:
recording and reporting information, epidemic emergency, cluster analysis, modelling disease
spread, and planning control strategies.
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Section 3
Risk assessment and management
Key contributions
Risk Management of game: from theory to practice
Milorad Radakovic1 and John Fletcher2
1Food Standards Agency, Aviation House, 125 Kingsway London, WC2B 6NH, United
Kingdom; milorad.radakovic@foodstandards.gsi.gov.uk
2Reediehill Farm, Auchtermuchty, Fife KY14 7HS, Scotland, United Kingdom
Summary
Risk Management is an integral part of Risk Analysis. It cannot be conducted in isolation from
Risk Assessment and Risk Communication. Although it is generally assumed that formal Risk
Management is carried out by government officials, in reality this need not be the case – risk
can be managed within the industry as long as the process is functionally separated from
Risk Assessment. The United Nations Food and Agriculture Organisation (FAO) and the
World Health Organisation (WHO) have jointly produced the Food Safety Risk Analysis ‘A
guide for national authorities’ (FAO/WHO, 2006), which provides a generic approach to Risk
Management and is essential reading for risk managers. Responsibility for Risk Management
lies with everyone in the food chain – in the case of game meat – ‘from forest to fork’. This
can include environmentalists, conservationists, hunters, food producers and processors,
transporters, retailers, consumers and officials. Live animals and game meat may carry a
great variety of biological, chemical and physical contaminants. These need to be judged
in the context of their significance and the risks they pose to those handling or consuming
game meat. At each stage of game meat production, every day, risks are managed by food
producers and official controllers fulfilling their legal obligations. There is no prescribed rule
as to when there is a need for formal Risk Management. Each case is different. Thus formal
risk analysis may be required to deal with an actual or perceived risk to consumers when, for
example, new emerging risks arise, or the evidence suggests that the current control measures
are either ineffective or not cost-effective. Risk managers face many challenges during this
process, of which perhaps the most difficult is ‘to translate’ and ‘define’ food safety issues in
easily understood language and to provide evidence based answers from which effective, yet
environmentally sustainable and proportionate measures can be developed.
Keywords: generic risk assessment, hazards, food handling, farmed game, wild game, United
Kingdom
1. Introduction
There are many definitions applicable to Risk Management in general and to game meat
production. These may be found in various dictionaries, in the Codex Alimentarius, in
European Commission literature and in the laws of respective countries. For the purpose of
this chapter consideration has been given to those found in the Codex Alimentarius (Box 1)
and within European Union (EU) legislation. Game is defined as either ‘wild’ or ‘farmed’ in
Regulation (EC) No. 853/2004 (EC, 2004):

• ‘wild game’ is defined as: ‘ungulates and lagomorphs, as well as other land mammals that
are hunted for human consumption and are considered to be wild under the applicable
law in the Member State concerned, including mammals living in enclosed territory under
conditions of freedom similar to those of wild game.’ It also includes ‘wild birds that are
hunted for human consumption’.
• ‘ farmed game’ includes ‘farmed ratites and farmed land mammals’.
Box 1. Definitions of Risk Analysis terms related to food safety, according to FAO/WHO Rome
2007 ‘Working Principles for Risk Analysis for Food Safety for Application by Governments’.
Hazard – A biological, chemical or physical agent in, or condition of, food with the potential to cause an
adverse health effect.
Risk – A function of the probability of an adverse health effect and the severity of that effect, consequential
to a hazard(s) in food.
Risk Analysis – A process consisting of three components: Risk Assessment, Risk Management and Risk
Communication.
Risk Assessment – A scientifically based process consisting of the following steps: (1) hazard identification;
(2) hazard characterisation; (3) exposure assessment and, (4) risk characterisation.
Risk Management – The process, distinct from Risk Assessment, of weighing policy alternatives, in
consultation with all interested parties, considering Risk Assessment and other factors relevant for the
health protection of consumers and for the promotion of fair trade practices, and, if needed, selecting
appropriate prevention and control options.
Risk Communication – The interactive exchange of information and opinions throughout the Risk Analysis
process concerning risk, risk-related factors and risk perceptions, among risk assessors, risk managers,
consumers, industry, the academic community and other interested parties, including the explanation of
Risk Assessment findings and the basis of Risk Management decisions.
Risk Assessment Policy – Documented guidelines on the choice of options and associated judgements for
their application at appropriate decision points in the Risk Assessment such that the scientific integrity of
the process is maintained.
Risk Profile – The description of the food safety problem and its context.
Risk Characterisation – The qualitative and/or quantitative estimation, including attendant uncertainties,
of the probability of occurrence and severity of known or potential adverse health effects in a given
population based on hazard identification, hazard characterisation and exposure assessment.
Risk Estimate – The quantitative estimation of risk resulting from risk characterisation.
Hazard Identification – The identification of biological, chemical, and physical agents capable of causing
adverse health effects and which may be present in a particular food or group of foods.
Hazard Characterisation – The qualitative and/or quantitative evaluation of the nature of the adverse
health effects associated with biological, chemical and physical agents which may be present in food. For
chemical agents, a dose-response assessment should be performed. For biological or physical agents, a
dose-response assessment should be performed if the data are obtainable.
Dose-Response Assessment – The determination of the relationship between the magnitude of exposure
(dose) to a chemical, biological or physical agent and the severity and/or frequency of associated adverse
health effects (response).

Exposure Assessment – The qualitative and/or quantitative evaluation of the likely intake of biological,
chemical, and physical agents via food as well as exposures from other sources if relevant.
Food Safety Objective (FSO) – The maximum frequency and/or concentration of a hazard in a food at the
time of consumption that provides or contributes to the appropriate level of protection (ALOP).
Performance Criterion (PC) – The effect in frequency and/or concentration of a hazard in a food that must
be achieved by the application of one or more control measures to provide or contribute to a PO or an FSO.
Performance Objective (PO) – The maximum frequency and/or concentration of a hazard in a food at a
specified step in the food chain before the time of consumption that provides or contributes to an FSO or
ALOP, as applicable.
This chapter, in very broad terms, describes the main risks to the handlers and consumers
which may arise from handling and consumption of game meat, the theory and practical
means of managing those risks and the roles and responsibilities of all concerned. Its purpose
is to provide an overview of the main hazards posed by game meat to humans as well as some
practical suggestions for managing those risks. It does not intentionally go into the details
of particular hazards, their prevalence in live animals and meat. The chapter also provides
the non-exhaustive list of simple questions that will need answers when conducting Risk
Management activities bearing in mind that each case is different.
Although not within the remit of this chapter, risk managers should be aware of their role in
providing food which is not only free from contamination but also nutritionally safe. Game
meats are recommended as preferable to conventional livestock by, for example, the World
Cancer Research Fund and the American Institute for Cancer Research who state that ‘if
eaten at all red meat to provide less than 10% of total energy but consumption of meat from
non-domesticated animals is preferable.’
2. Hazards in game meat (wild and farmed)
2.1 Which hazards are relevant in game meat?
Hazards here are divided into biological, chemical and physical hazards. Theoretically, there
may be many hazards in or on live animals or game meat and this needs to be assessed as to
the risk they pose to the public either through handling or eating game meat. Some examples
of hazards are:
• Biological: Salmonella spp., Campylobacter spp., Clostridia spp., Listeria, entero-
haemorrhagic Escherichia coli O157, Staphylococcus aureus, Yersinia enterocolitica,
Chlamydia spp., Francisella tularensis, Brucella spp., Mycobacterium bovis/avium,
aflatoxin, some viruses (e.g. avian influenza), yeasts, fungi, parasites (e.g. Trichinella,
Toxoplasma gondii), etc.
• Chemical: residues of antimicrobials, antiparasitics, pesticides, heavy metals (e.g. lead,
cadmium, mercury, etc.), other environmental chemicals (e.g. dioxins), hormones or
hormone like substances, mycotoxins, etc. and radioactive isotopes.
• Physical such as foreign bodies (bone, glass, metal, plastic, etc.).

2.2 How do we know about hazards?
The majority of hazards are invisible and may cause no damage to the animal but sometimes
they will cause observable symptoms and/or lesions. This is particularly true for the major
biological contaminants causing food poisoning, e.g. Salmonella, Campylobacter, or chemical
contaminants such as lead, mercury, etc. In many cases the hazards may not be obvious and
we need to rely on reports, surveillance data, animal and human data, and other sources of
data, e.g. from the environment.
2.3 How do hazards enter the live animal?
This depends on the type of hazard, whether it is biological, chemical or physical. Animals
are exposed to risk through direct and indirect contact with the environment which may
include equipment, other animals (including vectors) and interaction with humans or human
activities, e.g. feeding. Whilst this may be accidental, in some cases, as for example in the
irresponsible disposal of chemical waste to the environment, it may approach the deliberate
introduction of hazards to live animals.
There are many factors which may influence whether or not contaminants may harm animals.
This, for example in the case of microbial hazards, will depend on many interrelated factors
such as virulence, pathogenicity, pathway and susceptibility of animals. Exposure to chemical
hazards usually but not invariably involves ingestion, as for example in the case of game being
shot when physical hazards (pellets, dirt, etc.) are introduced to live animals.
Because wild animals graze and browse freely and may not be killed until many years old they
are especially susceptible to contamination from heavy metals such as cadmium, and from
radioactive fallout such as that which was released following the accident at Chernobyl. These
may accumulate during the lifetime of the animal to reach levels deemed toxic.
2.4 How do hazards enter meat?
Chemical hazards such as lead and mercury may be accumulated in offal, e.g. liver, and
kidneys, and to a lesser extent in meat, while animals are alive. Physical hazards are usually
introduced when animals are shot.
Biological hazards may be present in the meat before shooting as in parasites such as
Trichinella. Bacteria are normally present in the intestines or on the skin of animals and may
be introduced onto or into the meat during the shooting, or at the first stage of dressing and
evisceration in the field. In small wild game it is also possible that hazards may be introduced
from the mouth of dogs retrieving the game.
In all subsequent stages of the production cycle – transport, storage, dressing, cutting and
processing – bacteria or fungi may contaminate the surface of the meat. The survival and
further multiplication of the microbes will depend on the availability of food, temperature
and water activity. Bacteria and fungi may also contaminate meat during food preparation
in the kitchen.

3. Risk Management: from theory to practice
3.1 Risk Management principles
Risk and Risk Management terms are defined by WHO/FAO as follows:

• ‘Risk: a function of the probability of an adverse health effect and the severity of that effect,
consequential to a hazard(s) in food.’
• ‘Risk Management: the process, distinct from Risk Assessment, of weighing policy
alternatives, in consultation with all interested parties, considering Risk Assessment and
other factors relevant for the health protection of consumers and for the promotion of fair
trade practices, and, if needed, selecting appropriate prevention and control.’
The Codex Alimentarius Commission paper (CAC, 2007) provides ‘Working Principles for
Risk Analysis for Food Safety for Application by Governments’. They advocate applying the
following principles when managing risk:
1. National government decisions on Risk Management, including sanitary measures
taken, should have as their primary objective the protection of the health of consumers.
Unjustified differences in the measures selected to address similar risks in different
situations should be avoided.
2. Risk Management should follow a structured approach including preliminary Risk
Management activities, evaluation of Risk Management options, implementation,
monitoring and review of the decision taken.
3. The decisions should be based on Risk Assessment, and should be proportionate to the
assessed risk, taking into account, where appropriate, other legitimate factors relevant for
the health protection of consumers and for the promotion of fair practices in food trade,
in accordance with the Criteria for the Consideration of the Other Factors Referred to in
the Second Statement of Principles as they relate to decisions at the national level. National
Governments should base their sanitary measures on Codex Standards and related texts,
where available.
4. In achieving agreed outcomes, Risk Management should take into account relevant
production, storage and handling practices used throughout the food chain including
traditional practices, methods of analysis, sampling and inspection, feasibility of
enforcement and compliance, and the prevalence of specific adverse health effects.
5. Risk Management should take into account the economic consequences and the feasibility
of Risk Management options.
6. The Risk Management process should be transparent, consistent and fully documented.
Decisions on Risk Management should be documented so as to facilitate a wider
understanding of the Risk Management process by all interested parties.
7. The outcome of the preliminary Risk Management activities and the Risk Assessment
should be combined with the evaluation of available Risk Management options in order
to reach a decision on management of the risk.
8. Risk Management options should be assessed in terms of the scope and purpose of Risk
Analysis and the level of consumer health protection they achieve. The option of not taking
any action should also be considered.
9. Risk Management should ensure transparency and consistency in the decision-making
process in all cases. Examination of the full range of Risk Management options should,

as far as possible, take into account an assessment of their potential advantages and
disadvantages. When making a choice among different Risk Management options, which
are equally effective in protecting the health of the consumer, national governments
should seek and take into consideration the potential impact of such measures on trade
and select measures that are no more restrictive than necessary.
10. Risk Management should be a continuing process that takes into account all newly
generated data in the evaluation and review of Risk Management decisions. The relevance,
effectiveness, and impacts of Risk Management decisions and their implementation
should be regularly monitored and the decisions and/or their implementation reviewed
as necessary.
3.2 Generic Risk Management framework
Risk Management, in general terms, is described in the Food Safety Risk Analysis ‘A Guide for
national authorities’ (FAO/WHO, 2006). The generic Risk Management Framework (RMF) is
a structured process for food safety regulators consisting of four major phases and numerous
specific activities (Figure 1). The four phases are:
1. Preliminary Risk Management activities.
2. Identification and selection of Risk Management options.
3. Implementation of Risk Management decision.
4. Monitoring and review.
Preliminary Risk Management activities

t Identify food safety issues
t Develop risk profile
t Establish goals of Risk Management
t Decide on need for Risk Management
t Establish Risk Assessment, if necessary
t Consider results of Risk Assessment
t Rank risks, if necessary
Monitoring and review Identification and selection of Risk

Management options
t Monitor outcomes of control(s) t Identify possible options
t Review control(s) where indicated t Evaluate options
t Select preferred option(s)
Implementation of Risk Management decision

t Validate control (s) where necessary
t Implement selected control(s)
t Verify implementation
Figure 1. Generic framework for Risk Management (FAO/WHO, 2006).

Most stages of Risk Management cannot be seen in isolation and will require extensive
communication, coordination and collaboration between risk managers, risk assessors and
stakeholders. Whilst the responsibility for formal Risk Management usually lies with the
managers of the national food safety authority, in practice managers from industry may
serve as risk managers.
Each formal Risk Management case is different. An approach to each case will depend on
many factors such as the nature of the hazard, the actual and/or perceived risks to consumers,
the economic situation, etc. Throughout the Risk Analysis process the risk managers need to
make many subjective judgements and find answers to a number of specific questions. Under
each heading below there are examples of some simple questions that will need answers.
However, on a case-by-case basis there may be many more questions. Transparent Risk
Communication with all stakeholders starts at the beginning of the process and lasts till the
end of Risk Analysis. With increased awareness of the importance of minimising ‘carbon
dioxide footprint’, Risk Management must always assess environmental sustainability in
evaluating what is proportionate.
3.2.1 Preliminary Risk Management tasks
Identify and describe the food safety issue – What is the problem? What is an issue? Why
is it necessary to manage the risk? Develop the risk profile – What do we know about the
problem so far? What evidence is available? What is the situation? How are handlers and
consumers exposed to the hazards? What are the possible risks? What are the actual and
the perceived risks? What are consumers’ perceptions? What are current control measures?
How effective are current control measures? Establish the goals of Risk Management – What
needs to be achieved in broad terms? Decide on the need for Risk Assessment – Is formal
Risk Assessment needed? Why is formal Risk Assessment needed? How much will it cost?
Establish the Risk Assessment policy – What needs to be assessed? Will it be quantitative
or qualitative Risk Assessment? Commission a Risk Assessment, if necessary – Who will
make the Risk Assessment? How will the independence of the risk assessor be judged? Will
risk assessors be functionally independent? Consider the results of Risk Assessment – What
are the strengths/weaknesses of Risk Assessment? What are the uncertainties raised in Risk
Assessment? Is there any (immediate/short/long term) action required? Rank the risks, if
necessary – What is the relative importance of a risk?
3.2.2 Identification and selection of Risk Management options
Identify the possible options – What needs to be done to control risks? Is immediate/short/
long term action required? Evaluate the options – What is the effectiveness, including cost
effectiveness, of the different options in achieving objectives? Who can conduct the Risk
Management options? Is any action justified? Select the preferred option(s) – Which options
are effective and cost effective?

3.2.3 Implementation of Risk Management decision
Validate the control(s) where necessary – Will controls achieve the objectives that are set up?
Who will do the validation? Implement the selected control(s) – Who will implement the
chosen options? Verify implementation – How is it implemented?
3.2.4 Monitoring and review
Monitor outcomes of control(s) – What evidence exists to support the effectiveness of chosen
options? Review control(s) where indicated – Are the controls effective? Would new Risk
Assessment be useful?
4. Wild and farmed game meat production
4.1 Wild game meat production
Wild game meat may be sold directly to the final consumer by the hunter or through the shops
and supermarkets. It is estimated that in the EU over 50% of wild game meat goes through
an unapproved food chain with no official veterinary meat inspection being carried out.
Initial post mortem examination is performed in the field by a trained hunter who must sign
a declaration about any abnormal behaviour that may have been observed before killing and
relevant post mortem findings. It is an EU legal requirement that hunters must be trained in
health and hygiene, so they must have sufficient knowledge of the pathology of wild game to
undertake initial examination of wild game on the spot.
Each wild game species has ‘seasons’ within which they can legally be hunted. The British
Association for Shooting & Conservation (BASC) lists the hunting seasons for the species
found in the UK (see http://www.basc.org.uk//en/departments/game-and-gamekeeping/
game-shooting/shooting-seasons.cfm). The vast majority of wild game animals are killed
by shooting, with the exception of some small wild game (rabbits and hares) which can be
trapped before being dispatched by a blow to the back of the head. Game birds, wild waterfowl
and small wild game are shot using cartridges that contain a number of small pellets (some
may be lead pellets although substitutes are being introduced, especially for waterfowl),
whereas large wild game, e.g. wild deer and wild boar, are shot with a bullet. Deer are generally
gralloched (eviscerated) on the hill by trained hunters and an initial post mortem inspection
carried out by trained hunters who will sign a declaration that will accompany the carcase to
the processing plant where final veterinary post mortem inspection will take place.
There is variation in the time between animals being killed and their arrival at the plant for
processing. Animals are generally shot 24 hours before being transported for processing,
but on occasions the carcases may be held for several days in refrigerated facilities at the
hunting ground before transportation to the processing plants. This period will depend on
the remoteness of the hunting ground and the proximity of game handling establishments.
It would be environmentally unsustainable and uneconomic to send a refrigerated vehicle
many miles for small numbers of game carcasses. Carcasses should be suspended during

transport, however game bird carcasses and waterfowl carcasses may be laid down on vehicle
floors provided they are not warm. They should not be heaped up to a great depth. After killing
and arrival at the processing plant the carcases are chilled within a reasonable period of time:
large wild game to less than 7 °C and small wild game to less than 4 °C.
Traditionally in Britain wild game birds, as well as hares and deer, may be ‘hung’ in well
ventilated refrigerated spaces for a period of several days to tenderise the meat and enhance
the flavour. Game birds are then usually de-feathered ‘dry’ using plucking machines, although
some processing plants may immerse duck or geese carcases in hot wax to aid feather removal.
Carcases may be eviscerated manually, or by machines which act by sucking out intestines. If
breast meat only is required, as for example is commonly the case with woodpigeons, birds
are not de-feathered prior to removal of the meat.
Large wild game (deer and boar) are usually skinned at the plant and most of them require
trimming due to contamination during shooting. When animals have been poorly shot in
the abdominal area this can result in excessive contamination and total condemnation of the
carcase. In approved processing plants post mortem inspection of wild game carcases and
available offal are carried out as per regulation.
4.2 Farmed deer meat production
Farmed deer are usually killed at the place of rearing. Carcases are processed and meat sold
to the consumer through an established route similar to that of conventional domesticated
livestock. There is no hunting season; farmed deer can be killed throughout the year. Ante
mortem inspection is carried out by a veterinarian within 72 hours of death and where the
deer are to be killed in a field by free bullet this is best conducted by observing the entire
herd at rest. Farmed deer may be killed in the field by a free bullet to the head or upper neck.
They may also be brought into handling facilities or a conventional abattoir for stunning
with a captive bolt to the frontal region and bleeding. Meat inspection is carried out by
qualified meat inspectors and the premises are audited by qualified veterinarians. Farmed
deer meat must be cut and processed in approved cutting plants in the same way as the meat
of conventional livestock.
5. Practical Risk Management in game meat
5.1 An assessment of hazards and risks from game meat
Farmed game meat production in the UK has been, for many decades, subject to veterinary
official controls. However, prior to 2006, the majority of wild game meat produced for the
UK domestic market was not subject to veterinary inspection, with the exception of some
sold by larger retailers. As a part of the hygiene package the EU was, at that time, proposing
three legal measures (later adopted) relevant to wild game meat production.
1. The training of hunters.
2. Audited Hazard Analysis and Critical Control Points (HACCP), and
3. Official post mortem inspection of all.

The UK Food Standards Agency (FSA) wished to assess, in particular, the necessity for
veterinary presence and post mortem inspection, over and above the fully implemented
controls on general hygiene (Good Hygiene Practices (GHP) and HACCP) and the removal
of obviously unfit carcases by non-veterinary staff. To enable such an assessment, the FSA
requested a qualitative Risk Assessment to address the following risk question:
Under current UK law, what is the risk to human health (particularly of human
infection with a foodborne pathogen) from the handling/consumption of wild game
meat and how would the currently proposed EU hygiene proposal affect the risk ?
A qualitative Risk Assessment of hazards in wild game was made in 2003 (Coburn et al.,
2005). It summarises the risk from the identified hazards and concludes that veterinary post
mortem inspection alone will not significantly decrease the risk in the majority of cases as
long as an effective application of procedures based on HACCP principles and removal of
obviously unfit carcases takes place.
Different factors affect the risk from each hazard and these have been discussed within the
Risk Assessment. Hazards that have been identified as potentially harmful for human health
were those commonly found in different species: Salmonella spp., Campylobacter jejuni,
Escherichia coli O157, Clostridium botulinum, Yersinia pseudotuberculosis, Mycobacterium
avium/bovis and lead shot. Where data could not be identified, expert opinion has been
elicited and relevant data from other countries or species has been used.
Four categories of wild game were considered with potential risk pathways: (1) wild game
birds; (2) waterfowl; (3) large wild game; and (4) small wild game. Regardless of different
species each pathway has the same processing structure and similar risks (Figure 2). An
assessment of consumers’ kitchen hygiene and malpractices has also been made (Table 1).
RISK via Live wild animal

handling
Game larder
RISK via Transport and

handling processing
Storage, RISK via

RISK via
handling distribution and handling
retail
RISK via
handling and Preparation and
consumption consumption
Figure 2. Live wild animal-to-fork pathway for wild game animals.

Table 1. Incidence of food handling malpractices (adapted from Worsfold and Griffith, 1997)
Preparation stage Behaviour % Occurrences Result
Transport Transported food subject to temperature 45 Possible growth of

abuse organism
Refrigerated storage Chilled ingredients stored above 5 °C 58
Handling and preparing Hands not washed prior to preparation 66 Possible cross-
raw foods contamination and
Hands not washed after handling raw animal 58 contamination of
ingredients hands
Meat/poultry packaging not removed from 18
preparation area
Raw poultry washed in sink 33
All ingredients cut on a single board 60
Failure to use recommended method of 25
cleaning cutting board
Cooking Product not cooked to internal temperature 15 Possible survival of
of at least 74 °C organism
Post cooking handling Cooked product held at ambient temperature 35 Possible growth of
for more than 90 minutes organism
Cooked food cut on board not cleaned 8 Possible cross-
according to recommended method contamination and
Cooked food handled directly 9 contamination of
hands
Cooked product stored in refrigerator at a 17 Possible survival of
temperature above 5 °C organism
Product not reheated to an internal 11 Possible survival and
temperature of at least 74 °C growth of organism
Product reheated more than once, with 6
intervening periods at room temperature
In general, if the organism is present there is plenty of opportunity for growth and cross-
contamination, and particularly contamination of hands. The percentage of cooking steps
that fail to heat food adequately is also not negligible; therefore if the organism is present there
is a low, but not negligible probability that it remains viable. This coupled with the probable
growth and cross-contamination of organism if present, means that it is quite likely to grow
and spread.

6. Conclusions
There are potentially a number of risks associated with handling and eating game. An
assessment of the most obvious bacterial species and lead found that there was a low risk
to human health from the handling and consumption of game species in the UK. Proper
handling and cooking of game meat will destroy most microbial organisms; however, this
is not always the case. Every day Risk Management in game meat production is achieved by
all involved ‘from forest to fork’. Food producers are legally obliged to apply Good Hygiene
Practices (GHP) and procedures based on Hazard Analysis and Critical Control Points
(HACCP) principles continuously, properly and proportionately. Official controllers, on the
other hand, are required to perform auditing and inspection tasks (ante and post mortem
inspection, when appropriate) in order to verify food producers’ legal obligations. There is
also guidance available to help everyone in the food chain to manage risks appropriately on a
daily basis. More comprehensive general guidance, from WHO/FAO, is available for managers
when conducting formal Risk Management activities.
It is important to acknowledge that handling and/or consumption of game meat may also
result in human exposure to novel diseases. The knowledge about these agents or diseases is
sparse in wild game meat species. It is also true that more research and data is needed to assess
the risk from handling and consuming game meat outside approved food chain and from
different species. Risk Management poses many challenges for risk managers, risk assessors
and risk communicators, of which perhaps the most difficult is ‘to translate’ and ‘define’ food
safety issues in easily understood language and to provide evidence based answers from which
effective and proportionate measures can be developed. This can only be done by people who
are properly trained.
Each Risk Management case is different and must be done in a systematic well documented and
transparent way. It cannot be done in isolation from Risk Assessment and Risk Communication.
An approach to each Risk Management case will primarily depend on whether it is necessary
to deal with: (1) an actual and/or perceived risk to handlers or consumers; (2) new unknown
emerging risks; and (3) known risks where the evidence suggests that the current measures are
not effective in controlling the risks or are unnecessary. More training, better collaboration
and exchange of information is required from all involved handling and consuming game
‘from forest to fork’.

Acknowledgements
The authors would like to thank Ms Abrar Jaffer, Scientific Officer in Hygiene and Microbiology
Division of Food Standards Agency for her contribution in providing necessary reading
material, especially about hazards in game meat.
References
CAC (Codex Alimentarius Commission), 2007. ‘Working principles for Risk Analysis for food safety for application
by governments’. CAC/GL 62. Available at: ftp://ftp.fao.org/codex/Publications/Booklets/Risk/Risk_EN_FR_
ES.pdf. Accessed: 4 February 2010. ISSN: 1020-2579.
Coburn, H.L., Snary, E.L., Kelly, L.A. and Wooldridge, M., 2005. A Qualitative Risk Assessment of the hazards
and risks from wild game. Vet Rec. 157(11), 321-322. Available at: Accessed 4 February 2010. http://www.food.
gov.uk/science/research/researchinfo/foodborneillness/meathygieneresearch/m01prog/m01list/m01025/.
EC (European Commission), 2004. Regulation (EC) No. 853/2004 of the European Parliament and the Council of
29th of April 2004 containing specific rules for the hygiene of foodstuffs. Brussels, O.J. L 139/55.
FAO/WHO, 2006. Food safety Risk Analysis. Guide for national food safety Authorities. Available at: http://www.
who.int/foodsafety/publications/micro/riskanalysis06.pdf. Accessed: 4 February 2010. ISSN 0254-4725.
Worsfold, D. and Griffith C., 1997. Food safety behaviour in the home. British Food J. 99, 97-104.

The monitoring of selected zoonotic diseases of
wildlife in Lombardy and Emilia-Romagna, northern
Italy
Simone Magnino1, Matteo Frasnelli2, Massimo Fabbi3, Alessandro Bianchi4,
Maria Grazia Zanoni5, Giuseppe Merialdi6, Maria Ludovica Pacciarini7 and
Alessandra Gaffuri8
Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna (IZSLER)
‘Bruno Ubertini’
1Sezione di Pavia, Strada Campeggi 61, 27100 Pavia, Italy;
simone.magnino@izsler.it
2Sezione di Ravenna, Via del Limite 2, 48022 Lugo di Romagna, Italy
3National Reference Laboratory for Tularaemia, Strada Campeggi 61, 27100 Pavia, Italy
4Sezione di Sondrio, Via Bormio 30, 23100 Sondrio, Italy
5Central Diagnostic Laboratory, Via Bianchi 7/9, 25124 Brescia, Italy
6Sezione di Bologna, Via Fiorini 5, 40127 Bologna, Italy
7 National Reference Laboratory for Tuberculosis by Mycobacterium bovis, Via Bianchi 7/9,
25124 Brescia, Italy

8Sezione di Bergamo, Via Rovelli 53, 24125 Bergamo, Italy
Summary
Several zoonotic agents that infect wildlife may be transmitted to humans through
contaminated game meat. Toxoplasma gondii and Trichinella spp. infect the muscular tissue
at some stage of their biological cycle and consequently contaminate meat, while enteric
bacteria (e.g. Salmonella spp., verocytotoxic Escherichia coli (VTEC), Yersinia enterocolitica)
may contaminate the carcase after the animal’s death when the intestine is ruptured by shot
pellets or during evisceration. Gutting and skinning of carcases may lead to contamination
of humans with Mycobacterium bovis, Brucella spp., and Francisella tularensis, the etiological
agents of bovine tuberculosis, brucellosis and tularaemia, respectively. We report here the
occurrence of the above mentioned organisms over a 5-year period in some species of wild
mammals of Lombardy and Emilia-Romagna, northern Italy. Trichinellae were recovered
in 0.15% and 0.13% of samples from red foxes and wild boars, respectively. Antibodies to T.
gondii were detected in a percentage ranging from 23.3% to 33.3% of wild ruminants and
wild boars, and the parasite was also directly detected in European hares. Several serotypes of
Salmonella spp. were recovered from cervids, red foxes and wild boars with prevalences from
1.46% to 18.7%. Faecal samples from roe deer tested negative for VTEC, while Y. enterocolitica
was detected in about 8% samples from roe deer and from a few wild boars. M. bovis was
rarely recovered from red deer and red foxes, and occasionally from wild boar, where M.
microti, another species belonging to the Mycobacterium tuberculosis complex, occurred
much more frequently. Antibodies to Brucella spp. were very rarely detected only in European
hares, while F. tularensis was detected by PCR in about 7% European hares. This monitoring

Simone Magnino et al.
programme was conceived and implemented by the regional and local administrations in
close collaboration with the official veterinary services and the hunters’ associations.
Keywords: wild mammals, infectious disease, parasitic disease, zoonosis
1. Introduction
In the last few years, wildlife health has progressively become a cause for common concern
for different stakeholders, including veterinary services, public administrations, wildlife
conservationists, hunters, gamekeepers, farmers and for the whole community as well.
In Europe, changes in agricultural land use and in wildlife management practices have
recently and dramatically influenced the population dynamics of wildlife, often leading to an
overabundance of species (e.g. wild boars and roe deer) in several areas, which may in turn
impact on the health status of the populations (Gortázar et al., 2006).
Wild animals naturally harbour a number of agents of infectious and parasitic diseases which
may be transmitted to livestock and humans (Simpson, 2002; Bengis et al., 2004; Kruse et al.,
2004; Böhm et al., 2007). Transmission of diseases from wildlife to livestock and vice versa is
related to several variables, such as animal movements, restocking and environmental changes,
that lead to increased contact opportunities between animal populations, and allow pathogenic
organisms to cross species barriers. The risk of exchange of pathogens and transmission of
diseases between domestic and wild animals increases significantly when contacts occur in
common grazing areas, in open-air livestock rearing conditions and in organic livestock
production systems, and can be also mediated by vectors (Lanfranchi et al., 2003; Gortázar et
al., 2007; Kijlstra et al., 2009). Climate change also influences the development of infectious
and parasitic diseases in free-ranging animals by affecting the distribution of vectors and the
migratory routes of wildlife (Gilbert et al., 2008). As to the relevance of wildlife as a source of
disease for humans, it should be noted that about 60% of emerging infectious diseases globally
detected in human populations between 1940 and 2004 were zoonotic, i.e. acquired from
animals and about 70% of them originated from a wildlife reservoir (Jones et al., 2008). As a
consequence, the interest for the detection of pathogens in wildlife has considerably grown in
the last few years, leading to the development of disease monitoring programmes in several
European countries (Artois et al., 2001; Mörner et al., 2002).
In Italy, infectious and parasitic diseases of wildlife are monitored through laboratory testing
carried out by different institutions. Among them, departments from several universities and
a network of region-based governmental laboratories (the Istituti Zooprofilattici Sperimentali)
are prominently involved. The National Reference Centre for Wildlife Diseases (Ce.R.M.A.S.),
based in Aosta, northwestern Italy, provides the coordination of the activities of the regional
laboratories and the transmission of data of the wildlife health monitoring programmes
to the World Organisation for Animal Health (OIE). In Lombardy and Emilia-Romagna,
two regions in northern Italy, the monitoring of diseases of wildlife has been systematically
carried out for more than 10 years, and is based on the collection of data on the occurrence
of infectious and parasitic diseases which directly impact the health status of the animal
populations, or are shared between livestock and wildlife, or are zoonotic.

The monitoring of selected zoonotic diseases of wildlife in Lombardy and Emilia-Romagna
In this contribution, we present data on selected zoonotic diseases that can be transmitted
from wildlife to humans through contact with contaminated game meat either by ingestion
or handling during gutting and skinning procedures preliminary to consumption. Data
refer to the results of laboratory tests carried out through a five-year (2005-2009) monitoring
programme, and refer both to the direct detection of selected pathogenic organisms, as well
as to indirect tests for the detection of specific antibodies.
2. Study area
The data reported in this communication refer to wildlife of the territories of Lombardy and
Emilia-Romagna, two regions in northern Italy. Lombardy covers an area of 23,861 km 2
and is bordered by Switzerland to the north and by other Italian regions to the west, east
and south. Its northern part is a mountainous area limited by the Central Alps chain whose
altitude ranges from 1,000 to 4,020 m. Moving south, the mountains (40.6% of the territory)
slope into the hills (12.4%) and then to the plains of the Po Valley (47%). Emilia-Romagna is
a region located south of Lombardy and bordered on all sides by other Italian regions and by
the Adriatic Sea. The region covers an area of 22,124 km2 and approximately half of it, in its
northern part (48% of the territory), consists of the plains of the Po Valley, while the remaining
part of the region is covered by hills (27%) and mountains (25%), up to an elevation of 2,121 m.
The territories of both Lombardy and Emilia-Romagna are characterised by natural and semi-
natural habitats with different levels of human activities. Both regions are highly industrialised,
yet intensive farming is well developed. In addition, limited traditional farming activities are
still maintained in some marginal hilly and mountainous areas where, in summertime, large
flocks of sheep and goats are taken up to the alpine pastures.
Wildlife in the two regions include several species of wild ruminants, some of which – the
Alpine chamois and the Alpine ibex – are only found in the alpine area of Lombardy. Roe
deer and red deer are found in both regions, with a larger population of the former species
in Emilia-Romagna. Wild boar, red fox and several species of mustelids, lagomorphs, and
rodents are also widespread. In addition, the wolf (Canis lupus) and the bear (Ursus arctos)
have sporadically reappeared following reintroduction projects or for natural radiation. Due
to a combination of changes in agricultural land use, and in forest and wildlife management
practices, all populations of traditionally hunted wild mammals, particularly wild ungulates,
have significantly increased over the last twenty years to the sizes presented in Table 1. This
has been favoured by the reduction of traditional farming practices, which has made available
to wildlife vast areas formerly exploited by humans.
Wildlife disease monitoring programmes in the two regions were initially – around 1998 –
implemented by the official veterinary services at the request of the hunters’ association. Since
then, at the beginning of each year, the veterinary services in collaboration with the Istituto
Zooprofilattico Sperimentale della Lombardia e dell’Emilia-Romagna (IZSLER), the regional
governmental laboratory, have been planning the activities. Several zoonotic diseases are
included in the monitoring programmes, along with some livestock diseases for which wild
animals may act as a reservoir (e.g. classical swine fever and pseudorabies).

Table 1. Estimated population sizes (as to 2008) for some wild mammals in Lombardy and Emilia-
Romagna.
Species Region
Lombardy Emilia-Romagna
Roe deer (Capreolus capreolus) 26,000 90,000

Red deer (Cervus elaphus) 7,300 4,000
Alpine chamois (Rupicapra rupicapra rupicapra) 19,500 -
Alpine ibex (Capra ibex ibex) 2,600 -
Mouflon (Ovis musimon) 800 250
Wild boar (Sus scrofa) 3,300 (hunting bag/year) 17,000 (hunting bag/year)
Sources: Regione Lombardia, Direzione Generale Agricoltura [http://62.101.84.225/agrinet/fauna/

rapportofauna2008.htm]; Regione Emilia Romagna, Servizio Territorio Rurale [http://www.ermesagricoltura.it/
Box-Informazioni/Politiche-Faunistiche-e-Venatorie/Osservatorio-Faunistico-Venatorio/Dati/Ungulati].
Hunters are asked to collect samples (blood, faeces, swabs, and/or viscera) from animals
harvested during the regular hunting seasons, and both hunters and gamekeepers are
instructed to deliver the carcases of dead animals to the laboratory for collection of samples
at necropsy. In few cases, samples are also collected from injured animals temporarily hosted
in rehabilitation centres. Investigations are carried out in several departments of IZSLER for
the detection of organisms that infect the muscle tissue (Trichinella spp., Toxoplasma gondii),
or that may contaminate the carcases due to intestinal rupture (enteric bacteria: Salmonella
spp., verocytotoxic Escherichia coli (VTEC), Yersinia enterocolitica) or due to contact with
other infected tissues or organs during skinning and gutting (other bacteria: Francisella
tularensis, Brucella spp., Mycobacterium bovis). Bacteriological, parasitological and serological
examinations are carried out following the procedures recommended in the Manual of
Diagnostic Tests and Vaccines for Terrestrial Animals (OIE, 2009a) as summarised in Table 2.
All results are presented in Table 3.
3. Diseases
3.1 Trichinellosis
Trichinellosis is a zoonotic parasitic disease caused by nematodes of the genus Trichinella

that mainly infect carnivorous and omnivorus mammals with cannibalistic and scavenging
behaviour. Some trichinellae also infect birds and reptiles (Pozio, 2005). All Trichinella
species are transmitted directly by ingestion of parasitised muscle tissue and are maintained
in nature in sylvatic or domestic cycles, the latter mainly involving domestic pigs. Humans
are accidental hosts of trichinellae and are infected following consumption of contaminated
raw or undercooked meat (Pozio, 2000).

Table 2. Organisms considered and detection methods employed in the monitoring programme.
Organism Detection methods
Direct method Serological test
Trichinella spp. artificial digestion nd

Toxoplasma gondii PCR ELISA
Salmonella spp. culture, serotyping nd
Verocytotoxic Escherichia coli culture, PCR nd
Yersinia enterocolitica culture nd
Mycobacterium bovis PCR, culture nd
Brucella spp. PCR CFT, RBT
Francisella tularensis PCR, culture AT
Legenda: PCR = polymerase chain reaction; ELISA = enzyme-linked immunosorbent assay; AT = agglutination
test; CFT = complement fixation test; RBT = Rose Bengal test; nd = not done
In Europe, four species of trichinellae have been detected in wild and domestic animals: T.
spiralis, T. britovi, T. nativa and T. pseudospiralis. The first two species are the most widespread
and occur up to 60-61° latitude north, while T. nativa is only found in arctic and subarctic
regions, and limited data are available as to the overall distribution of T. pseudospiralis (Pozio
et al., 2009). T. britovi is known to occur in Italy in several wild species, including red foxes
and wild boars, and T. pseudospiralis has been detected in nocturnal birds of prey (Pozio et
al., 1999; Pozio, 2007).
According to Regulation (EC) No. 2075/2005 of the European Union legislation (EC, 2005),
carcases of farmed and wild animal species susceptible to Trichinella infestation shall be
systematically examined for trichinellae. Data from the monitoring programme in wildlife
is required when applying for Trichinella-free status for a pig herd (EC, 2005).
Our data refer to the testing of samples of muscular tissue collected from the diaphragm
and the forearm muscles, which are the predilection sites for the localisation of trichinellae
in the wild boar and red fox, respectively, and processed by artificial digestion according to
the EU regulation (EC, 2005).
Trichinellae were detected in the muscular tissue of several wild boars (n=36) sampled in
Lombardy in 2007 and 2008, and in a red fox and a wild boar sampled in Emilia-Romagna
in 2008 and 2009, respectively (Table 3). Typing was carried out by multiplex PCR (Pozio and
La Rosa, 2003) only for the latter two trichinellae, and confirmed the occurrence of T. britovi
in the fox, while the isolate from the wild boar was identified as T. pseudospiralis, a species
previously identified in wild boars of France, Finland, Sweden and the Netherlands, as well as
in other mammals and birds (Pozio and Zarlenga, 2005). The detection of T. britovi in the red
fox confirms the circulation of this Trichinella species in the study area (Pozio, 2007), while

Table 3. Test results of a 5-year wildlife disease monitoring programme for selected diseases in Lombardy and Emilia-Romagna.
228
Disease or infection Animal species Detection method1 Year Total
Prevalence % (95% CI)
2005 2006 2007 2008 2009
positive/tested positive/tested positive/tested positive/tested positive/tested
Trichinellosis wild boar AD 0/191 0/1,196 11/6,111 26/11,106 1/10,468 38/29,072

0.13 (0.09-0.18)
red fox AD - 0/21 0/87 1/204 0/336 1/648

0.15 (0.01-1.00)
Toxoplasmosis roe deer ELISA - - - - 122/460 122/460
26.52 (22.59-30.85)
wild boar ELISA 4/23 - 63/281 7/13 - 74/317
23.34 (18.88-28.47)
PCR - - 0/53 - - 0/53
0 (0.00-5.49)
European hare PCR 1/1 1/10 1/5 3/71 3/120 9/207
4.35 (2.14-8.36)
mouflon ELISA - - - - 5/19 5/19
26.31 (10.12-51.42)
red deer ELISA - - - - 5/15 5/15
33.33 (12.99-61.31)
Brucellosis wild boar CFT 0/152 0/162 0/433 0/307 0/192 0/1,246
0 (0.00-0.24)
roe deer RBT, CFT 0/224 0/24 0/136 0/13 0/142 0/539
0 (0.00-0.55)
PCR - - 0/19 - 0/58 0/77
0 (0.00-3.8)
European hare RBT, CFT 0/14 0/109 0/93 2/195 0/99 2/510
0.39 (0.06-1.29)
red deer CFT - 0/52 0/9 - 0/164 0/225
0 (0.00-1.32)
chamois CFT - 0/191 - - - 0/191
0 (0.00-1.55)

Salmonellosis wild boar C - 92/311 123/666 79/643 147/736 441/2,356
18.7 (17.41-20.10)
roe deer C - - 2/110 2/104 1/16 5/230
2.17 (0.8-5.2)
red deer C - - 1/67 0/36 2/102 3/205
1.46 (0.37-3.93)
red fox C - 0/17 2/17 - 0/3 2/37
5.13 (0.86-15.92)
chamois C - - - - 0/10 0/10
0 (0.00-25.88)

Infection by VTEC roe deer C, PCR - - 0/43 0/81 - 0/124
0 (0.00-2.38)
Yersiniosis by roe deer C 0/10 0/6 10/114 9/107 - 19/237
Y. enterocolitica 8.02 (5.02-12.43)
red deer C - 0/11 - - - 0/11
0 (0.00-23.84)
chamois C - 0/10 - - - 0/10
0 (0.00-25.88)
wild boar C - 4/5 - - - 4/5
80 (29.88-98.95)
Tularaemia European hare AT 8/2,394 12/2,658 15/1,693 2/1,980 20/2,376 57/11,101
0.51 (0.39-0.67)
PCR 8/134 0/91 0/43 3/34 8/92 19/260
7.31 (4.58-11.36)
wild boar AT 0/231 - - 0/14 7/372 7/617
1.13 (0.50-2.43)
PCR - - - - 0/35 0/35
0 (0.00-8.20)
roe deer AT 0/248 - - 0/22 0/312 0/582
0 (0.00-0.51
chamois AT - 0/85 - - - 0/85
0 (0.00-3.46)
1 Detection methods used: AD = artificial digestion; ELISA = enzyme-linked immunosorbent assay; PCR = polymerase chain reaction; CFT = complement fixation test; RBT
229
= Rose Bengal test; C = culture; AT = agglutination test.
the occurrence of T. pseudospiralis in the wild boar adds new information, as this species had
never been reported before in Italy in a mammalian host.
3.2 Toxoplasmosis
The protozoan parasite Toxoplasma gondii infects all warm-blooded animals (mammals and
birds). It multiplies asexually in several mammal and bird species, which are intermediate
hosts, and reproduces sexually only in domestic and wild felids, which act as definitive hosts.
In intermediate hosts, T. gondii forms tissue cysts that can be mainly found in neural and
muscular tissues, and are occasionally detected in visceral organs such as lung, liver, kidneys,
while definitive hosts shed the oocysts of the parasite in their faeces. Transplacental infections
may also occur in several animal species (Dubey and Beattie, 1988). Wild ruminants are
infected through ingestion of T. gondii oocysts shed by felids, which is the case also for the
wild boar that can be infected also through consumption of parasitised muscle tissue (Gauss
et al., 2005, 2006).
Antibodies to T. gondii in wildlife have been found in several surveys carried out in European
wildlife, with the highest levels of seropositivity and antibody titers in animals living in
mountainous areas with shade and high humidity, which are environmental conditions that
favour the survival of T. gondii oocysts (Gauss et al., 2006). In Europe, seropositivity to T.
gondii has been detected in all tested wild ruminant species, i.e. moose, reindeer, red deer,
roe deer, fallow deer, mouflon, ibex (Kapperud, 1978; Oksanen et al., 1997; Vikøren et al.,
2004; Aubert et al., 2006; Gaffuri et al., 2006; Gamarra et al., 2008). Antibodies have been
detected more often in roe deer compared to other cervids, with percentages of seropositivity
that tend to increase with the age of the sampled animals (Gauss et al., 2006). In addition,
antibodies have been often found in wild boars (Edelhofer et al., 1996; Hejlícek et al., 1997;
Lutz, 1997; Gauss et al., 2005; Bártová et al., 2006; Ruiz-Fons et al., 2006; Antolová et al.,
2007; Richomme et al., 2009).
Several sera collected in our study area from roe deer, wild boar, red deer and mouflon were
tested for specific antibodies to T. gondii with a commercial ELISA (ID Screen® Toxoplasmosis
Indirect ELISA, IDVET, Montpellier, France). An overall seropositivity ranging from 23.3%
to 33.3% was detected (Table 3). In order to further investigate the seropositivity of wild
boars and directly detect the parasite, a PCR-RFLP assay targeting the 18S small-subunit
ribosomal gene of T. gondii (Magnino et al., 1998) was subsequently carried out on samples
of heart tissue of a subset (n=53) of seropositive animals, but all samples tested negative. T.
gondii was also detected by PCR-RFLP in visceral samples of European hares, which are
highly susceptible to the infection and often die of acute fatal toxoplasmosis (Gustafsson et
al., 1988; Luppi et al., 2009).
The finding of antibodies to T. gondii in a significant proportion of samples collected from

several wild species suggests that the parasite is widespread in the study area. The risk of
infection through consumption of undercooked meat of T. gondii-infected game from the
area should be taken into account, as reports of human infection linked to consumption
of game meat have been published (Sacks et al., 1983; Ross et al., 2001). As to measures for
reducing the risk, thorough cooking (to 67 °C) as well as freezing (to -13 °C) of meat are

suitable treatments for ensuring the inactivation of T. gondii tissue cysts. In addition, since
evisceration and handling of T. gondii-infected game may also represent a risk of infection to
humans, hands and all materials coming in contact with uncooked meat should be washed
with soap and water for inactivating the parasite (Dubey and Beattie, 1988; Dubey, 1994).
3.3 Salmonellosis
Salmonellae are enteric bacteria which are able to infect a large number of animal hosts,
including all wild animals (Millán et al., 2004). Wild mammals can act as reservoirs or
carriers of different serotypes of Salmonella, including the ones that infect birds and cold-
blooded animals. Although animals are the major reservoir of Salmonellae, environmental
contamination from human activities, including livestock farming and waste disposal, is
likely to be an additional source of infection also for wildlife (Murray, 2000). Moreover, the
probability of being infected by Salmonella is also related to the feeding habits of each species,
which makes omnivores and carnivores more susceptible to the infection.
In our study area, most samples were collected from wild boars and to a lesser extent from
roe deer, red deer and red foxes. Caecal or rectal contents were either directly sampled by the
hunters or collected during necropsy and cultured according to standardised procedures for
the detection of Salmonellae (ISO 6579:2002 Annex D).
Salmonellae were often isolated from wild boars (Table 3) in a percentage of samples ranging
from 3.9% (Confidence Interval (CI), 2.76-5.67) to 26% (CI, 23.86-28.25), in relation to the
geographical area of sampling. Serotyping was performed according to the Kauffman-White
typing scheme. The most prevalent serotypes were S. Coeln (81 isolates), S. Typhimurium
(74), S. Ball (43), S. Thompson (37), S. Veneziana (37), S. Enteritidis (18), S. Infantis (5). In
addition, cultures yielded several (40) isolates of S. enterica subsp. diarizonae, a subspecies of
Salmonella that is typically found in reptiles and other cold-blooded animals.
Salmonellae were also occasionally detected in the intestinal content or faeces of roe deer,
red deer and red foxes (Table 3). As to the serotypes, S. Typhimurium, S. Napoli, S. enterica
subsp. enterica, S. Veneziana and S. Mishmarhaemek were detected in red deer and roe deer,
and S. Virchow in foxes.
The recovery of a large number of Salmonella isolates from wildlife, mainly from wild boars,
is related to the wide occurrence of these bacteria in the environment and in a number of
animal hosts. The high variability in the prevalence and serotypes detected in different
geographical areas of sampling could not be clearly referred to differences in contamination
sources from human activities, including livestock farming and waste disposal, and should
be further investigated.
3.4 Infection by verocytotoxic E. coli (VTEC)
Several serotypes of VTEC, including E. coli O157:H7, are associated with disease in humans,
with clinical presentations ranging from diarrhea to hemorrhagic colitis and hemolytic
uremic syndrome, a severe condition which may be fatal (Karmali et al., 2010).

Healthy ruminants, mainly cattle, harbour more than 400 different serotypes of VTEC in
their intestine, as part of their gut flora (Blanco et al., 2004) and shed the organism in their
faeces. VTEC have been occasionally detected also in wildlife, e.g. in faecal samples from
white-tailed deer (Odocoileus virginianus) in the United States (Rice et al., 1995; Sargeant et
al., 1999; Fischer et al., 2001; Renter et al., 2001; Dunn et al., 2004), moose (Alces alces) in
Canada (Todd et al., 1999), reindeer (Rangifer tarandus) in Norway and Finland (Lahti et al.,
2001; Aschfalk et al., 2003), sika deer (Cervus nippon) in Japan (Fukuyama et al., 1999), as
well as in the intestinal content of red deer and roe deer in Italy (Conedera et al., 2004) and in
rectal swabs from red deer in Spain (García-Sánchez et al., 2007). The occurrence of VTEC in
wildlife has been linked to transmission from farm animals through faecal contamination of
shared pastures (Keene et al., 1997; Sargeant et al., 1999). On the other hand, the importance
of wild animals for the transmission and/or persistence of VTEC within farms or between
farms is unknown (Nielsen et al., 2004).
Faecally-contaminated bovine foods and dairy products have been identified as important
vehicles of VTEC to humans in a number of outbreaks and sporadic cases of disease (Rangel
et al., 2005; Yoon and Hovde, 2008), and contaminated game meat as well has been linked to
human disease (Keene et al., 1997; Rabatsky-Ehr et al., 2002; Ahn et al., 2009). Game meat
from caribou (Rangifer tarandus), deer and roe deer has been occasionally found contaminated
with VTEC (Milley and Sekla, 1993; Thoms, 1999; Nagano et al., 2004; Lehmann et al., 2006),
and a recent study further indicates that game animals are an additional reservoir for human
pathogenic VTEC (Miko et al., 2009).
Our data refer to the testing of a population of roe deer in the province of Reggio Emilia,
Emilia-Romagna (Spaggiari et al., 2009). Faecal samples were examined for VTEC by selective
culture and PCR assays targeting intimin (eae) and verotoxin (VT) genes (García-Sánchez et
al., 2007). Fifteen out of 124 (12.1%) E. coli isolates tested positive for the eae gene, but none
of them carried VT genes.
Failure to detect VTEC in the study area might have been related to the small number of
tested samples, as well as to the restriction of testing to one species only (roe deer). Previous
surveys in other wildlife populations either failed to detect VTEC as well (Wahlström et al.,
2003; Lillehaug et al., 2005) or detected low prevalences, ranging from 0.3% in roe deer in
northeastern Italy (Conedera et al., 2004) to 1.5% in red deer in Spain (García-Sánchez et al.,
2007) to 0.25%, 0.6% and 2.4% in different surveys in white-tailed deer populations in the
United States (Sargeant et al., 1999; Fischer et al., 2001; Renter et al., 2001).
3.5 Yersiniosis by Y. enterocolitica
Y. enterocolitica is an enteric bacterium associated with foodborne disease in humans

(Bottone, 1999). The organism has been detected in wildlife, e.g. in Norway in 29/2,243
(1.3%) faecal samples from reindeer (Kemper et al., 2006) and in 8/170 (4.7%) faecal samples
of red deer (Aschfalk et al., 2008), in the rectal content of 41/131 (31%) wild boars from Japan
(Hayashidani et al., 2002) and from the Canton of Geneva in Switzerland, with a significantly
higher positivity detected in tonsil samples than faecal samples (Fredriksson-Ahomaa et al.,

2009). Recent surveys also detected Y. enterocolitica on the surface of game meat in Bavaria,
Germany (Bucher et al., 2008).
In our survey, caecal content samples from wild ruminants and wild boars were tested for
Yersinia spp. by the cultural method, employing selective media and special procedures,
including cold enrichment. Typing was carried out according to the procedure described
by Bottone (1999). Overall, Y. enterocolitica was detected in a limited percentage (7%) of
samples, while other species (Y. bercovieri, Y. kristensenii, Y. frederiksenii, Y. intermedia) were
detected more frequently (13% of samples). All isolates were recovered from wild boars and
were typed as biotype 1A, which includes Y. enterocolitica strains that are considered non-
pathogenic, although recent evidence suggests that some of them can actually be virulent
(Tennant et al., 2003).
3.6 Tuberculosis
Tuberculosis (TB) by M. bovis is a disease that occurs worldwide in a wide range of domestic
and wild animals (Michel et al., 2010). Affected wildlife may show no clinical signs of disease,
or just exhibit a loss of weight. The occurrence of TB in wildlife may interfere with eradication
programmes of the disease in livestock, and may also pose a direct risk to human health (De
Lisle et al., 2002).
M. bovis belongs to the Mycobacterium tuberculosis complex (MtbC), which includes M.

tuberculosis, M. caprae, M. microti and other species. Humans may get infected with M. bovis
and the other species belonging to MtbC following direct contact with infected animals,
mainly through inhalation of contaminated aerosols, and consumption of milk. Meat
consumption has not been documented as a way of transmission of MtbC to humans, but
direct contact with organs and tissues of infected animals during skinning and evisceration
procedures might lead to infection by hand-to-mouth contact (De la Rua-Domenech, 2006).
Samples collected in Lombardy from wildlife were submitted for isolation of mycobacteria
and for detection and differentiation of MtbC. Retropharyngeal and submandibular lymph
nodes were collected from carcases of wild boar (n=4,200), red deer (n=208), roe deer (n=152)
and red foxes (n=53) and examined by macroscopical inspection. Upon detection of TB-like
lesions consisting in small rounded, necrotic and often calcified foci, samples of tissue were
decontaminated and cultured for mycobacteria in a solid medium (Lowenstein-Jensen-ST)
and a liquid culture system (BACTEC ™ MGIT ™ 960; Becton, Dickinson and Company).
Samples were concurrently processed for the detection of MtbC by DNA extraction and
PCR assay targeting the IS6110, an insertion element (IS) present in the genome of the MtbC
group (Haddad et al., 2004). In case of growth, cultures were identified by molecular and
biochemical tests (Boniotti et al., 2009). Raw samples that tested positive with the IS6110 PCR
but negative by culture were retrieved and processed for the direct identification of MtbC by
a PCR-restriction fragment length polymorphism assay targeting the gyrB gene (gyrB PCR-
RFLP), which allows the differentiation of MtbC species (Niemann et al., 2000). Typing of
mycobacteria strains was carried out by spoligotyping, a method based on PCR amplification
of a highly polymorphic direct repeat locus (DR) (Kamerbeek et al., 1997) and by analysis of
the VNTR loci as described by Boniotti et al. (2009).

A limited percentage (333/4,200; 7.9%) of wild boar lymph nodes showed macroscopic lesions
consistent with TB infection; out of these samples, 228 were positive to IS6110 PCR but only
180 were confirmed by gyrB PCR-RFLP. Surprisingly, in most samples (n=170) the main
etiological agent was identified as M. microti, a mycobacterial species that is commonly
isolated from rodents (mice and voles) and has been also detected in larger mammals such
as cats, pigs and llamas as well as in humans (Van Soolingen et al., 1998; Xavier Emmanuel
et al., 2007), while M. bovis was only detected in 10 samples. Cultures yielded 20 isolates of
M. microti and 4 isolates of M. bovis. Genetic profiles of all M. bovis isolates were found to
differ from those of isolates recovered in recent outbreaks of TB in cattle from the same areas
(Pacciarini et al., 2006).
Although no macroscopic lesions were found in the lymph nodes of red deer, roe deer and
red foxes, it was decided to analyse all samples by both IS6110 PCR and culture. Few samples
tested positive at PCR, namely 7/152 from roe deer, 5/208 from red deer, and 3/53 from red
foxes. Subsequent analysis by gyrB PCR-RFLP confirmed the detection of M. microti in the
red deer (n=2/208) and roe deer (n=1/152), and of M. bovis in the red deer (n=3/208) and red
foxes (n=2/53), but no isolate could be recovered in culture.
Overall, our data do not suggest a significant role for wild mammals as a reservoir for bovine
tuberculosis in the study area.
3.7 Brucellosis
Brucellosis is a disease caused by bacteria of the genus Brucella, that currently includes nine
species, namely B. abortus, B. melitensis, B. suis, B. canis, B. ovis, B. neotomae, B. pinnipedialis,
B. ceti and B. microti (Whatmore, 2010). The first three mentioned species are the agents of
bovine brucellosis, small ruminant brucellosis and swine brucellosis, respectively, which
are diseases that cause huge economic losses and important morbidity in domestic and wild
animals. Brucellae are transmitted to humans either by direct contact with infected animals,
or by ingestion of contaminated animal products, usually milk and dairy products, while
inhalation of contaminated aerosols is an important route of infection for occupationally
exposed people, such as workers in laboratories and slaughterhouses (Seleem et al., 2010).
In wildlife, B. abortus and B. suis have been isolated in a number of species, such as bison
(Bison bison), deer, wild boar, African buffalo (Syncerus caffer), European brown hare, with
the infection being endemic in some of these species worldwide (Godfroid, 2002). As to
European wildlife, B. melitensis has been recovered sporadically from chamois and ibex
in the Italian and French Alps and its occurrence has been interpreted as a spill-over from
infected small ruminants (Garin-Bastuji et al., 1990; Ferroglio et al., 1998). In addition, B.
suis is known to occur in Central Europe in the wild boar and in the European hare, which
are both considered to be reservoir of B. suis biovar 2 (Garin-Bastuji et al., 2000; Godfroid,
2002; Al Dahouk et al., 2005). High values of seropositivity to brucellae and the isolation of
B. suis biovar 2 have been also recently reported in wild boars in Piedmont, a region west of
Lombardy (Bergagna et al., 2009).

In our study area, antibodies to brucellae were not detected in wild boars nor in wild
ruminants. To this regard, it should be noted that brucellosis was eradicated from livestock
of the study area in 1996. In addition, in a serological survey carried out between 1995 and
2002 in roe deer and chamois in the central Italian Alps in Lombardy, all samples tested
negative for antibodies to brucellae (Gaffuri et al., 2006). Antibodies to brucellae were only
detected in the European hare, where two samples tested positive (Table 3) by the Rose Bengal
test (RBT) and complement fixation test (CFT).
As to direct testing by real-time PCR (Bogdanovich et al., 2004), no sample from any species
was positive for brucellae. The low (0.39%) seropositivity detected in hares suggests a very
limited circulation of brucellae in this species. Nonetheless, since there is a risk of introduction
of B. suis with imported hares from Eastern Europe, monitoring for brucellosis in the two
regions is set to continue both in the European hare and the wild boar, which are the main
reservoir hosts of B. suis.
3.8 Tularaemia
Francisella tularensis is the etiological agent of tularaemia, a zoonotic bacterial disease that
mainly affects rodents (mice, voles) and lagomorphs (hare, rabbit), and occasionally other
mammals and birds (Foley and Nieto, 2010; Padeshki et al., 2010). Currently, four subspecies
of F. tularensis that differ in virulence and geographical distribution are recognised, namely:
tularensis, holarctica, mediaasiatica and novicida. F. tularensis subsp. tularensis (type A),
which is more virulent, is found in North America, while F. tularensis subsp. holarctica
(type B) is less virulent and occurs in the Old World and occasionally in North America
and F. tularensis subsp. mediaasiatica is moderately virulent and has been only isolated
in Central Asia. F. tularensis subsp. novicida has low virulence in humans, and has been
isolated from clinical cases only in a few occasions (Johansson et al., 2000). F. tularensis
is listed among the most dangerous organisms (also known as ‘Category A agents’) that
can be used for bioterrorism purposes, due its low infectious dose, ease of dissemination
and high case-fatality rate (Davis, 2004). The organism can be transmitted to humans by
direct contact with infected animals (mostly hares), by tick or insect bite, and by exposure
to contaminated aerosols, e.g. to aerosols generated when grass is cut in areas contaminated
with carcases of infected animals (Feldman et al., 2001; Foley and Nieto, 2010). Outbreaks of
human disease may also follow the ingestion of contaminated water as occurred in the past
in several countries, including Bulgaria (Christova et al., 2004), Norway (Brantsaeter et al.,
2007), Turkey (Willke et al., 2009), and also recently in Tuscany, Italy (Fabbi et al., 2009).
Although the transmission of F. tularensis following ingestion of meat is considered as

exceptional, humans may get infected preliminary to the preparation and consumption of
meat, i.e. when skinning and gutting contaminated carcases, as also recently reported in
outbreaks of disease occurred in Germany, Spain and France (Hofstetter et al., 2006; Schätzle
and Schwenk, 2008; Mailles et al., 2009; Aldea-Mansilla et al., 2010). The Oltrepo’ Pavese, an
hilly area in the province of Pavia, in the southern part of Lombardy, has been an endemic
area for tularaemia since the 1960s, when the first documented cases of disease in Italy were
reported both in hares (Rinaldi et al., 1964) and humans (Bianchi, 1966).

During the monitoring activity, F. tularensis was occasionally detected in European hares
by two PCR assays, one that allows the identification of the genus Francisella and the other
specific for F. tularensis subsp. holarctica, and by culture on special media enriched with
cysteine (Forsman et al., 1994; Johansson et al., 2000; Foley and Nieto, 2010). The infection was
demonstrated both in indigenous hares found dead in Lombardy in the province of Pavia and
in Emilia-Romagna in the provinces of Piacenza, Parma and Bologna, and in hares imported
from Hungary, Slovakia and Romania for restocking. All isolates were typed as F. tularensis
subsp. holarctica. The organism was detected by PCR in Emilia-Romagna also in a few coypus
(Myocastor coypus), which might be frequently exposed to water-borne infections due to their
habit of living in burrows along river banks. Overall, antibodies to F. tularensis were very
rarely detected in indigenous hares, which is probably due to the short course and high fatality
of the disease. On the other hand, high antibody titers (1:640 and higher) were occasionally
found in a few imported hares from Eastern Europe, where the disease is endemic. In addition,
antibodies to F. tularensis were detected in a few sera collected from wild boars.
It should be noted that the handling and especially gutting and skinning of hare carcases
may be associated to a risk of infection by F. tularensis, due to the occurrence of the organism
in hares in the two regions. The presence of F. tularensis in the environment, as well as in
vectors and wildlife, is often revealed when cases of disease are diagnosed in humans, as
recently reported in Italy (Castro et al., 1999), Czech Republic (Cerný, 2001), Spain (Martín et
al., 2007; Allue et al., 2008), Germany (Kaysser et al., 2008) and France (Mailles et al., 2009).
Tularaemia has been diagnosed also recently in hares in several European countries, including
Austria (Hofer, 2002), Germany (Müller et al., 2007), France (Mailles et al., 2009) and Spain
(Aldea-Mansilla et al., 2010). Direct methods for detecting and monitoring tularaemia in the
European hare should be preferred to serological methods as mortality rate in this species
is often high and death occurs before the development of a detectable antibody response.
4. Conclusions
The encroachment of humans and domestic animals into wildlife habitats and the growing
interest in and popularity of wildlife has globally led to enhanced monitoring and increased
detection of diseases in wild animals (Rhyan and Spraker, 2010). In this regard, the monitoring
and surveillance of wildlife for the prevention and control of diseases is considered by OIE
‘a crucial component of the safeguarding of global animal and public health and related
agriculture and trade issues’ (OIE, 2009b).
The wildlife disease monitoring programme that has been established in Lombardy and
Emilia-Romagna has been conceived and implemented by the regional and provincial
administrations and the official veterinary services in close collaboration with major partners
in the field, i.e. hunters and gamekeepers. Over the years, the programme has grown to
become an ongoing system for assessing trends of infectious and parasitic diseases of wildife
and has allowed documenting the occurrence of most zoonotic pathogens in several wild
species of Lombardy and Emilia-Romagna.

In spite of the commitment of hunters and gamekeepers, the quality of a few samples was
affected due to a failure to comply with the given instructions regarding the choice and
range of tissues and organs to be submitted for analysis. In addition, inconsistent handling
and poor storage and transport conditions of the samples occasionally led to their extensive
contamination or to inactivation/loss of viability of the associated infectious organisms. As
a consequence, a few samples were either deemed unfit for the laboratory tests or yielded
questionable results, which precluded their inclusion in the analysis.
The focus of the monitoring programme needs to be expanded in the future in order to
tackle emerging zoonotic infections. For this purpose, specific instructions in the collection
of samples and building of technical capacity at the laboratory level will have to be provided.
In addition, training courses for hunters and gamekeepers will need to reflect any relevant
update in the epidemiology of zoonotic diseases that may be acquired from wildlife.
Special attention will also need to be given to the harmonisation of the sampling schemes
and laboratory methods with other monitoring programmes implemented at regional and
national levels, as that will allow the direct comparison of data referred to different wildlife
populations.
Acknowledgements
We thank the regional and provincial offices charged with the management of wildlife
(Assessorati alla caccia e pesca), the official veterinary services and the hunters’ associations
of Lombardy and Emilia-Romagna for the collaboration in planning and implementing the
monitoring programme.
We thank Dr Edoardo Pozio from the Community Reference Laboratory for Parasites of the
Istituto Superiore di Sanità in Rome for the typing of trichinellae, and Dr Mario D’Incau from
the Laboratory of Bacteriology of IZSLER in Brescia for the typing of Salmonellae.
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Assurance of food safety along the game meat
production chain: inspection of meat from wild
game and education of official veterinarians and
‘trained persons’ in Austria
Rudolf Winkelmayer1, Peter-Vitus Stangl2 and Peter Paulsen3
1Vet. Adminstration of Lower Austria/District Bruck a.d.L./NÖ, Fischamenderstraße 10,
2460 Bruck an der Leitha, Austria; rudolf.winkelmayer@noel.gv.at
2Ministry of Health, Radetzkystraße 2, 1031 Vienna, Austria
3Institute of Meat Hygiene, Meat Technology and Food Science, Department for Farm Animals

1210 Vienna, Austria
Summary
Wild game harvested by common hunting methods represents a sustainable source of meat.
Most hunting practices fulfil animal welfare requirements and the meat has usually a high
nutritive value and favourable sensory characteristics. The mode of production, however,
differs from that of farmed animals, and prompts taking specific measures along the food
chain. Among these measures, meat inspection and hygiene control are necessary to assure
that meat from wild game is ‘safe’ for the consumer. In 1994, Austria implemented an
inspection system in compliance with EU directive 92/45/EEC (EC, 1992), which involves
three categories (hunter, trained person and official veterinarians), and provides continuous
training and evaluation of these people in a consistent and logical way. With the ‘new’ food
hygiene legislation entered into force in 2006, only minor adaptations had to be made. In
compliance with EU recommendations, ‘Guides to Good Practice’ form an important element
not only in training of inspection personnel, but they also serve as references when game
carcasses or food businesses trading meat from wild game are inspected.
Keywords: official veterinarian, trained person, wild game, Trichinella
1. Introduction
Meat from wild game is a highly valued product with favourable composition and sensory
characteristics (Hoffman and Wiklund, 2006). It is also attractive for consumers who are
concerned about ethical and sustainable aspects of food production, similar to products from
organic farming (Winkelmayer and Paulsen, 2008a). Careful inspection of game before killing
and post mortem examination of carcass and organs, as well as the strict adherence to certain
rules of good hygiene practice along the food chain (‘from forest/field to fork’) are necessary
to assure that all quality traits are preserved and that game meat is not a source of hazards
to the consumer. Although the quantity of meat produced by hunting is small compared to

pork or beef, it is estimated that in Austria the total carcass weight (eviscerated, skin-on) of
wild game is ca. 9 million kg/year, see Table 1.
In this field of meat production, self-checks form an essential part of food safety. In Austria,
considerable effort is spent on training and motivation of hunters and trained persons, also
by providing continuous education and voluntary evaluation schemes. This system allows the
risk-based use of (limited) inspection capacities of the competent authority.
In comparison with farm animals, meat from wild game constitutes a minor fraction of the
average meat supply. However, this does not mean that the inspection of wild game should be
less strict than that of farm animals. Based on a calculation scheme given by Fehlhaber (2007),
the number of consumers possibly exposed when a hazardous game carcass is not examined
at all or passes all examinations and is processed into portions can be estimated. Calculations
were done based on 2007 data from a major Lower Austrian game-handling establishment.
Thus, roe deer (15.3 kg, uneviscerated, skin-on; average of 18,000 carcasses), red deer (87.8 kg,
average of 2,900 carcasses) and wild boar (44 kg, average of 3,600 carcasses) would yield 37,
265 and 147 portions, respectively (Winkelmayer and Paulsen, 2008b). In the most extreme
case the number of affected consumers could be as high as the number of portions. This is a
clear indication that each single game carcass has to undergo thorough inspection in order
to minimise health hazards to the consumer (Table 2).
2. Inspection system for wild game in Austria (excl. Trichinella)
2.1 Pre-1994 situation
Before 1994, no specific regulations existed in Austria with respect to the inspection of wild
game. The Food Act of 1975 stated that hazardous or spoiled food must not be placed on the
market. The Austrian Food Codex, Chapter B14 (Anonymous, 2010) required that game meat
must undergo a veterinary meat inspection before being processed to meat products, which
was practically only possible for meat from farmed game.
Table 1. Quantity of wild game1 harvested in Austria in 2007 (Pontasch, 2008).
Species Animals (ca.)
Roe deer 260,000

Red deer 48,000
Chamois 22,000
Wild boar 25,000
Brown hare 180,000
Pheasant 160,000
Wild duck 70,000
1 Represents a total carcass weight of ca. 9,000,000 kg; the average game meat consumption is ca. 0.5 kg/person/year.

Assurance of food safety along the game meat production chain
Table 2. Average number portions/servings of meat and meat products per carcass (serving sizes according
to Fehlhaber, 2007).
Species Average Meat cuts Meat for Number of portions Max.

carcass weight in kg processing number of
(skin-on)1 in kg Meat cuts Meat products Total consumers
in kg
Red deer 88 40 3 235 30 265 265

Wild boar 44 20 3 117 30 147 147
Roe deer 15.5 5.5 0.5 32 5 37 37
1 Rounded to 0.5 kg.
At EU level, a directive of 1992 (EC, 1992) laid down the principles for the inspection and
hygiene of meat from wild game, with the exception of marketing of carcasses or meat cuts
from the hunter to the final consumer or to local food businesses which supply directly to
the consumer.
2.2 Development from 1994 to 2004/2006
Austria became member of the European Ecomonic Area in 1994 (member of the European
Union in 1995), and hence, had to implement Directive 92/45/EC (EC, 1992). While this
caused no substantial problems with respect to hygiene requirements for game larders and
game-handling establishments, mandatory game meat inspection presented a challenge.
The inspection scheme had not only to meet EU directive requirements for placing safe
game meat on the market, but there were numbers of additional requirements and practical
considerations:
1. Common hunting strategies in Austria: the majority of large game is harvested by
still hunting, outside typical working hours or at weekends. Hunting grounds can be
remote from settlements and also quite distant from (the relatively few) game-handling
establishments. Carcasses are delivered to game larders (i.e. cooling facilities shared by
a number of hunting grounds), from where they are picked up at regular intervals by
wholesalers.
2. Hunting traditions: offals, e.g. liver are traditionally consumed after the hunt and would
thus not be available for a veterinary inspection at game larders or at game-handling
establishments.
3. Availability and costs of inspection personnel: inspection of wild game immediately after
the hunt or at game larders exclusively by meat-inspecting veterinarians was not a realistic
option with respect to both costs and limited capacities.
4. Applicability for all branches of marketing: ante and basics steps of post mortem inspection
should be identical for all branches of marketing, from local ‘direct’ marketing, trading
to approved game-handling establishments, intra-community-trade and export to third
countries. A modular scheme (see below) was found most suitable for that purpose.
5. Compliance of hunters and game handling establishments.

As a consequence, a three-step inspection scheme was designed (Anonymous, 2004), where

hunters have to provide information on the ante mortem condition of hunted game and on
carcass and stomach and intestines upon evisceration. ‘Auxiliaries for inspection of game meat’,
i.e. registered hunters with special training – now termed 'trained persons' – were responsible for
inspection of the carcass and edible inner organs at game larders. The examination by hunters
as well as trained persons was based on a checklist (Table 3). When carcasses were delivered to
game-handling establishments or serious abnormalities detected by hunters or trained persons,
carcass and organs had to be presented to the official veterinarian for inspection (Figure 1 and 2).
Table 3. Inspection criteria for wild game in Austria, 1994-2004 (after Winkelmayer et al., 1994, based
on Austrian Game Meat Regulation 1994).
To check ante mortem Examples of abnormalities
Nutritional condition • emaciation

Behaviour, movement • dazed state, somnolence
• lameness
• complicated fractures
Skin, hair • hair loss
• hair discolouration
Orifices • exudates (blood, foam)
• diarrhoea
Atypical sounds • cough
• groan/rattle
To check post mortem Examples of serious abnormalities
Outer surface, orifices, • emaciation

nutritional condition • wounds, when numerous or inflamed
• tumours and abscesses, when numerous
• anal region soiled by massive diarrhoea
• compound fractures, when not caused by shooting
• omphalitis
• orchitis
• swollen joint(s) (arthritis)
Thoracal and abdominal • foreign matter, esp. gut contents or urine (not when caused by shooting), when
cavities pleura or peritoneum are discoloured
• pus and other septic masses
• signs of acute pleuritis or peritonitis
Organs and pleura/ • considerable changes in: colour, consistency, odour
peritoneum • tumours and abscesses, when numerous or disseminated
• abnormities of liver (with exception of parasites) and spleen
• considerable gas formation in guts esp. when organs are discoloured (greening)
• enteritis (serositis, peritonitis)
Visible muscle surfaces • considerable changes in: colour, consistency, odour
• tumours and abscesses, when numerous or disseminated

Inspection before shooting
Shooting
Evisceration (within 3 hrs. past

shooting) and inspection of
carcass, stomach, intestines
No serious abnormities1? Serious abnormities1? Contact official

Certification – ‘A’ side of a veterinarian
double-sided tag affixed to
carcass
Distribution: Eviscerated, ‘skin- supply to consumer: Game Meat Regulation does

on’, chilled2, not frozen apply only to Trichinella3
Intended for distribution to game-handling

establishment?
Intended for distribution to local Inspection4 of carcass, lung, heart, liver, spleen
retailer (butcher), restaurant? obligatory within 36 hrs. (eg by trained person at
game larders; or done by official veterinarians
doing inspections at wholesaler)
Certification – ‘B’ side of a double-sided tag
affixed to carcass
Inspection3 of carcass and Delivery of carcass without viscera to game-

spleen by trained person or handling establishment, if no serious abnormities
official veterinarian obligatory detected
within 36 hrs. Certification –
‘B’ side of a double-sided tag
affixed to carcass
Delivery of carcass without Certificate of the inspection (Side ‘A’ and ‘B’)
viscera to local food supplier, if and carcass controlled by official veterinarian
no serious abnormities detected
Figure 1. Meat inspection of wild ungulates (adopted from Paulsen and Winkelmayer, 2000) in Austria
1994-2004/2006.
1 see Table 3.
2 Storage of eviscerated, hide-on ungulates at -1 to +1 °C max. 15 days, +1 to +7 °C max. 7 days.
3 Microscopical examination of wild boar meat for Trichinella sp. obligatory. Can be performed by trained person
with additional education. For this type of Trichinella examination, 5 specified muscles are sampled (diaphragm,
intercostal muscle, leg muscle, tongue, M. masseter) and a total of 14 subpieces is inspected via trichinoscopic method.
4 Examination of wild boar meat for Trichinella sp. by digestion method obligatory

Inspection before shooting
Shooting
Evisceration When abnormities1:

(recommended, but not official veterinarian
obligatory
Direct to consumer, Skin-on / feather-on, Certificate for each

retail shop, restaurant chilled2 species of small game
shot on one day and one
region
To game-handling
establishment
Inspection by official
veterinarian
Figure 2. Meat inspection of small game (i.e. birds, rabbit, hare) in Austria, 1994-2004/2006 (adopted
from Paulsen and Winkelmayer, 2000).
1 see Table 3.
2 storage at -1 to +4 °C max. 15 days.
2.3 Current system
2.3.1 Role of trained persons
Only minor modifications were necessary to ensure compliance with the ‘new’ EU ‘hygiene
package’. The inspection system involves three categories of personnel (Table 4), and their
involvement depends on:
• the way of marketing; and
• the presence of certain abnormalities detected during ante and post mortem inspection; or
• the suspicion of environmental contamination.
In 2008, there were ca. 110,000 licensed hunters in Austria, ca. 25,000 of which were registered
as trained persons.
For education and training, a series of textbooks exist and they provide the basis for uniform
and consistent training courses in Austria. The basic training (mandatory for all hunters to
pass the exam for the first hunting license) includes diseases of wild game and good hygiene
practice during primary production (evisceration, cooling, and storage of the carcass) and is
addressed in the ‘Österreichischer Jagdprüfungsbehelf’ (a guide to preparation for hunter´s
examination in Austria; Sternath, 2006).
For trained persons, a textbook on game meat inspection (‘Wildbrethygiene’; Winkelmayer

et al., 2008) and one on hygiene during primary processing and fresh meat production

Table 4. Ways of marketing of meat from wild game and involved inspection personnel1.
hunter (Step 1) trained person (Step 2) official veterinarian (Step 3)
Self-supply - - **
Local trade2 + + **
Intra-community trade + + +
Export + + +
1 Inspection: - = none; + = mandatory; ** = only in case of serious abnormities, as defined in Regulation (EC) No.
854/2004, Annex 1, Sect. IV, A, Par. 3., lett. (a), (d), (e).
2 i.e. direct supply to final consumers or to food businesses supplying directly to the final consumer (local, small
quantities)
(‘Wildbretdirektvermarktung’; Winkelmayer et al., 2007) are provided. The latter textbook

also serves as a ‘Guide to Good Practice’ for processing of game meat.
Training of official veterinarians in hygiene of meat from wild game is done in the course of
the periodical ‘evaluation’, with training material provided by the Federal Ministry of Health
(‘Food Safety Module D’).
Notably, all abovementioned training materials originate from the same team of authors,
which contributes to consistency in contents as well as presentation. This consistency forms
an important part of the ‘forest/field to fork’ principle.
2.3.2 Examination performed by the official veterinarian
A simple sensory examination will give a first indication if spoilage has begun, and more
specific techniques as known from meat inspection of slaughter animals can be applied, if
deemed necessary. Details for the examination are defined in Regulation (EC) No. 854/2004,
Annex 1, Sect. IV, A (EC, 2004a) and additional national legal texts ensure uniformity in
inspection procedures.
The official veterinarian also considers the information on the ante mortem condition of the
game and on abnormalities detected during evisceration. This information is provided on a
double sided tag (‘Wildfleischanhänger’) affixed to large game carcasses or on an accompanying
document (‘Begleitschein’) for small game. It is the responsibility of the hunter to document
the ante mortem inspection results and abnormalities detected during evisceration (i.e.
on the carcass and the intestines), whereas the examination by the trained person focuses
on the carcass and the (edible) inner organs. This document also assures traceability in
the food chain. When game carcasses are delivered to game handling establishments with
missing or incomplete certificates, these carcasses will not pass the examination of the trained
veterinarian.

2.3.3 Legal framework according to marketing branch
In principle, Regulations (EC) No. 178/2002 (EC, 2002), No. 852/2004 (EC, 2004b), No.
853/2004 (EC, 2004c) and No. 854/2004 (EC, 2004a) apply to the production of meat from
wild game. Additional Austrian legislation comprises a food safety act (Anonymous, 2006a),
acts on meat inspection, an act on direct marketing of certain foods (Anonymous, 2006b)
and on the local trade of certain meat products (Anonymous, 2006c) and a number of either
binding texts (e.g. on labelling of food, personal hygiene) or recommendations.
Regulation (EC) No. 853/2004 (EC, 2004b) does neither apply to domestic use of game meat
by the hunter nor to small quantities that are traded locally either directly to the consumer
or local food businesses (butchers, retailers, restaurants, canteen, etc.) supplying directly
to the consumer. This sector of ‘direct marketing’ is governed by national legislation (e.g.
Anonymous, 2006b). Table 5 gives an overview on relevant legal texts.
3. Trichinella inspection
3.1 Epidemiological situation
The epidemiological situation in Austria has been reviewed by Duscher et al. (2005). In brief,
the red fox is a wildlife reservoir for Trichinella britovi, and reservoirs are mainly located
in Western provinces. All reported cases of human trichinellosis (incidence <0.1/100,000
inhabitants/year) originate from foods consumed outside Austria or self-imported food.
Table 5. Legal texts relevant for production and processing meat from wild game1.
Reg. (EC) No. 852-854/2004
Reg. (EC) No. 178/2002
Lebensmittel-Direkt-
(Anonymous, 2006b)
(Anonymous, 2006a)
vermarktungsVO
EinzelhandelsVO
(EC, 2004a,b,c)
Lebensmittel-
(EC, 2002)
Marketing
Meat from large and small Own private use - - - -

game
Carcasses, eviscerated, skin- Local, direct, small quantities + - + -
on; not deep-frozen
Carcasses, skinned, meat cuts Local, direct, small quantities + 852/2004 + -
Meat products Local, direct, small quantities + 852/2004 + +
853/2004
1 - = not applicable; + = applies.

The last autochtonous case of trichinellosis in a domestic pig dates from before 1970. In wild
boar, there are only sporadic cases, mostly with not well-documented history, at a rate of ca. 1
case/10 years. Ca. 50% of hunted wild boars are marketed via game-handling establishments
and tested by the digestion method. To date, Trichinella pseudospiralis has not been detected
in wild boars in Austria.
3.2 Inspection by official veterinarians
Trichinella testing of wild boars in game-handling establishments is done by official

veterinarians by digestion method according to Regulation (EC) No. 2075/2005 (EC, 2005).
3.3 Inspection by trained persons – Lower Austrian model
Regulation (EC) No. 2075/2005 (EC, 2005) does not apply to meat from wild boars supplied
from the hunter directly to the final consumer or to food businesses supplying directly to
the final consumer (‘direct marketing’). Federal legislation allows Trichinella testing of wild
boars destined for local trade by trichinoscopy done by specially educated trained persons.
The decision to allow this option was motivated by (a) the relatively stable epidemiological
situation and (b) the absence of Trichinella pseudospiralis.
This option has been, however, not implemented by all provinces. In addition to a primary
training course, voluntary evaluation schemes/proficiency tests are offered. If these possibilities
are not used, the competent authority will evaluate the trained person.
3.3.1 EU and national legislation
Lower Austria contributes ca. 2/3 to the hunting bag of wild boars in Austria. Not all carcasses
are collected and processed by game handling establishments, but a substantial fraction is
marketed by the hunters directly to the consumer or to local food retailers supplying directly
to the consumer (‘local trade’; Winkelmayer and Paulsen, 2008a).
With the exception of animals hunted for private use, wild boars undergo an ante and post
mortem inspection and an examination for Trichinella sp. For carcasses collected by game
handling establishments, EU law applies and these carcasses are tested by the digestion
method according to Regulation (EC) No. 2075/2005 (Stangl and Paulsen, 2005).
Inspection of carcasses for ‘local’ trade is regulated by national legislation. While the methods
in ante and post mortem inspection are identical to those applied for carcasses under EU
law, it is usually sufficient that the hunter and a trained person perform this examination.
With respect to Trichinella examination, wild boar carcasses entering a game handling
establishment must be examined by the digestion method.
As ‘direct marketing’ of wild boars, and thus Trichinella inspection for such carcasses, is not
covered by EU legislation, the Austrian provinces have been given the option to implement
Trichinella examination by trained persons using the trichinoscopic method. This option
exists since 1994. Minor adaptations were necessary to ensure compliance with Regulation

(EC) No. 2075/2005 (EC, 2005) (number of samples taken from the carcass and number of
examined subsamples). The restrictions laid down in Article 16 of this regulation apply (i.e.
max. 10 carcasses/day; digestion method not available; meat is marked with a health mark
that is clearly different from the common ‘oval’ health mark provided for in Article 5(1)(a) of
Regulation (EC) No. 853/2004 (EC, 2004b), and that the meat is not used for the production
of products where the production process does not kill Trichinella).
Lower Austria has taken this opportunity, and established a basic training course, and several
options for periodical (voluntary or mandatory) evaluation. If more than 10 carcasses are to be
examined by one trained person, or if the meat is to be used for products where the production
process does not kill Trichinella, then trained persons have to take samples and send them to
an accredited laboratory where Trichinella testing is done by the digestion method. Also, if
Trichinella is detected (irrespective of the method used), an alert procedure comes into force
which makes digestion method mandatory also for carcasses intended for ‘direct marketing’.
3.3.2 Requirements for trained persons performing Trichinella inspection
Trained persons already active in inspection of game can apply for additional training in
Trichinella inspection. After passing a basic training course, they have to register at the
veterinary district administration. Trained persons have to be evaluated by an official
veterinarian on a regular basis. In Lower Austria, the frequency and extent of such controls
will be based on evidence of successful attendance of advanced courses and proficiency tests
(Kahrer, 2009).
3.3.3 Basic training course
The basic training course is an obligatory one-day course and consists of theoretical parts (4
hours) and 2 practicals (4 hours).
In the first practical, the participants practice

• Working with the microscope.
• Trichinoscopic examination with meat containing inactivated larvae.
• Sampling of wild boar carcasses.
The second practical is a sort of proficiency test. Each participant has to examine 7 numbered
fields in a previoulsy prepared compression glass. The only information given to the participant
is that at least one field will contain trichinellae. The results are reported and in case of
discrepancies, the examination is, at the end of the training session, repeated together with
an experienced veterinarian. This step has been found to be very effective in improving the
relative sensitivity, and subsequently, relative accuracy of the trichinoscopic examination
(Table 6).
Based on the results, the competent authorities receive an annual summary report. Individual
result forms of the trichinoscopic examination are collected and can be presented to the
competent authority upon request.

Table 6. Efficacy of Trichinella examination by freshly trained (‘basic’) and experienced participants.
‘Basic’ ‘Experienced’
(497 samples tested by 71 participants) (695 samples tested by 174 participants)
Relative sensitivity, % 83.1 98.4

Relative specifity, % 93.1 94.4
Relative accuracy, % 88.1 96.5
(‘agreement’)
3.3.4 Advanced courses
Advanced courses are voluntary half-day events and can be attended only by trained persons
already registered for Trichinella examination. Participants are requested to bring their
microscope and all necessary tools with them.
The theoretical part consists of 1.5 hours (update on legislation and possibility for discussion
on practical and administrative issues); 2.5 hours are provided for practicals.
The first practical is the self-evaluation of the microscope according to a checklist addressing
completeness and functionality of equipment and the major mechanical or optical problems
related to microscopes used in a rugged environment. In case of unsatisfactory results, small
repairs or cleaning jobs are done directly in the course.
For the second practical, each participant receives four coded meat samples (1-4 of the samples
may contain inactivated Trichinella larvae) and has to examine the samples microscopically.
The results are reported and discrepancies are discussed.
Advanced courses started in 2008, with a total of 174 participants, and are repeated on an
annual basis, with a total capacity of 40-60 participants/year (i.e. 2 courses). Based on the
results, the competent authorities receive an annual summary report. The overall performance
is demonstrated in Table 6. Completed self-evaluation forms and results of the trichinoscopic
examination are collected and can be presented to the competent authority upon request.
3.3.5 Proficiency tests
The first proficiency test has been completed in 2001. Basically, each participant received
two coded samples and had to examine them and report the results (Paulsen et al., 2003). As
only two meat samples were distributed per participant, the power of this test was low (i.e.
the chance to report two correct results by simply guessing was 25%). On the other hand,
participants had not been informed about the test in advance, and 91% responded, which
indicated a very high compliance for a voluntary proficiency test.

Similar to the experiences in the training courses, the relative accuracy was 86.3% for the
first set of samples. When participants with discrepant results received a second sample set,
relative accuracy improved to 97.1%.
4. Conclusions
In 1994, Austria implemented an inspection system in compliance with EU directive 92/45/

EEC, which involves three categories (hunter, trained person (former: auxiliaries) and official
veterinarians), and provides continuous training and evaluation of these people in a consistent
and logical way. The Austrian game meat inspection system relies on motivated trained
persons and recognises the self-responsibility of the primary producers (i.e. the hunters).
With the ‘new’ food hygiene legislation of 2006, only minor adaptations had to be made.
This system is augmented by ‘Guides to Good Practice’. Grants are available from hunting
associations for research targeted to practical game meat hygiene issues.
The current game meat inspection scheme includes periodical re-training, but there is only
limited possibility to check if the routine activity of trained persons and hunters is always in
full compliance with legislation. Practically, routine control of the health certificates affixed to
large game carcasses and comparison to the findings on the carcass should give an indication
if trained persons perform correctly. This is done on a routine basis in game-handling
establishments and on a risk-based plan in game larders. However, a real re-evaluation
scheme under field conditions should be developed.
In the game food chain, the use of lures, baits, winter feeding, etc. has received too few
attention and the primary producers must be made more aware that they are responsible for
the safety and traceability of feedstuff used for wild game (Deutz, 2010).
Between EU member states, there are also some unresolved items:

• Currently, it is unclear under which provisions wild game game carcasses (skin-on) can
be transported directly from hunting grounds or from game larders to game-handling
establishments in other EU countries.
• In Austria, it is principally possible that trained persons, who have received their training
and examination in other EU countries, register in Austria. A harmonisation of education
schemes for trained persons in all EU countries is necessary.

Acknowledgements
This contribution is based on presentations given at the conference ‘Hygiene of wild game
meat and its safety within the food chain’, Prague, 22.-24.4. 2009, and on the 50. Arbeitstagung
des Arbeitsgebietes ‘Lebensmittelhygiene’ der DVG, Garmisch-Partenkirchen, 29.9.-2.10.2009.
References
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Anonymous, 2006b. Lebensmittel-Direktvermarktungsverordnung, BGBl. II No. 108/2006.
Anonymous, 2006c. Lebensmittel-Einzelhandelsverordnung, BGBl. II No. 92/2006.
Anonymous, 2010. Austrian Food Codex, 3rd ed., available at: www.bmg.gv.at.
Deutz, A., 2010. Aspekte der Lebensmittelsicherheit bei der Fütterung von Schalenwild in freier Wildbahn und in
Gehegen. Rundsch. Fleischhyg. Lebensmittelüberw. 62(1), 6-8.
Duscher, G., Winkelmayer, R. and Prosl, H., 2005. Schwarzwildverbreitung in Gebieten mit Trichinellenfunden
bei Füchsen in Österreich. Wien. Tierärztl. Mschr. 92, 315-321.
relating to the killing of wild game and the placing on the market of wild-game meat. O.J. L 268/35-53.
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28 January 2002 laying down the general principles and requirements of food law, establishing the European
Food Safety Authority and laying down procedures in matters of food safety. O.J. L 31/1-24.
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of 29 April 2004 laying down specific rules for the organisation of official controls on products of animal origin
intended for human consumption. O.J. L 226/83-127.
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of 29 April 2004 on the hygiene of foodstuffs. O.J. L 226/3-21.
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of 29 April 2004 laying down specific hygiene rules for the hygiene of foodstuffs. O.J. L 226/22-82.
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Other contributions

Structure and legal framework for the direct local
marketing of meat and meat products from wild
game in Austria: the Lower Austrian model
1210 Vienna, Austria; peter.paulsen@vetmeduni.ac.at
Summary
The ‘new’ EU food hygiene legislation prompted for an update of the Austrian legislation
on the marketing of meat or meat products from wild game from the hunter directly to
the consumer or local food suppliers. The Lower Austrian implementation is an integrated
model (‘forest-to-fork’, Winkelmayer, 2006) which recognises the self-responsibility of the
hunters supplying meat from wild game and which encourages this food sector to establish
an own codex of good practice (in the sense of a ‘guide to good practice’). Core elements are
a well-established educational and training concept, templates for documentation of good
manufacturing practice and an evaluation system including microbiological examination
of the products.
Keywords: wild game, meat, own-checks, direct local marketing, Good Hygiene Practice,
microbiology, forest-to-fork
1. Introduction
From official statistics, it can be estimated that in Austria 1/3 to 2/3 of the yearly hunting
bags are – either as fresh meat or as meat products – marketed from the hunter directly to the
consumer or to local food retail establishments (Kainz and Paulsen, 2005). It is conceivable
that the fraction of game meat traded via this ‘local marketing’ had been in the same order of
magnitude before a legal framework was established. This indicates that there must already
be considerable experience in handling of meat from wild game. As there are no records that
such meat or meat products had been involved in foodborne disease incidents, these empirical
practices seem to have provided a certain level of safety. The establishment of a legislation
specifically addressing this food sector started in 1994, and underwent a revision in 2006,
to ensure that direct marketing was regulated in compliance with the EU ‘hygiene package’.
Concurrently with this specific legislation, empirical meat handling practices have been
evaluated along the production chain ‘from forest-to-fork’ and adjusted to comply with
science- and risk-based food safety. This included, for example: location of the shot wound
(Winkelmayer et al., 2005); time to onset of cooling of eviscerated large game carcasses
(Paulsen and Winkelmayer, 2004), relation of visible contamination to the microbial

contamination of wild game carcasses (Paulsen et al., 2003), storage of uneviscerated pheasant
(Paulsen et al., 2008), application of Good Hygiene Practice (GHP) in cutting and deboning
of pheasants (El-Ghareeb et al., 2009), and optimising safety and quality traits in fermented
sausage from wild game (Bauer et al., 2007).
It is not surprising that most of the research work is funded by hunters associations, as these
associations have a vital interest that game meat is ‘safe’ food.
2. Current legal framework in Austria
Only parts of EU legislation apply to local marketing of fresh meat from wild game, most
relevant being Regulation (EC) No. 178/2002 (EC, 2002) and, when carcasses are broken down
into meat cuts, Regulation (EC) No. 852/2004 (EC, 2004a). Austrian legislation provides an act
on direct marketing. This act governs the hygienic conditions for direct and local marketing
of small quantities of certain primary products of vegetable (berries, mushrooms) or animal
origin, such as milk, eggs, carcasses of rabbit, poultry, wild game; in addition, this legislation
covers also meat cuts from wild game.
Said legislation is very ‘lean’, in the sense that only few binding limits are specified and it is
obvious that it appeals to the self-responsibility of the food business operators. For example,
the term ‘local’ is not explicitly defined, which means that ‘local market’ could be the entire
area of Austria; and small quantities are explicitly defined only for rabbits and poultry, which
are slaughtered at and sold directly from the farm (5,000 and 10,000 per year, respectively).
For meat products, another act sets parts of Regulation (EC) No. 853/2004 (EC, 2004b) into
force. The main items addressing wild game are given in Table 1.
3. Achievements to date
Lower Austria is the largest province in Austria, also in terms of contribution to the annual
hunting bag. The hunting association of Lower Austria, in cooperation with external expert
and the veterinary authority has developed a training and evaluation concept which is based
on four pillars:
1. Already existing infrastructure in terms of trained persons since 1994.
2. Training courses on meat processing with practical in approved cutting rooms under
veterinary supervision.
3. Checklists for self-checks and GHP compliance.
4. Microbiological own-checks.
3.1 Good Hygiene Practice (GHP) and documentation
Sector-specific ‘Guides to Good Practice’ are suggested in EU food hygiene legislation for
the different branches of food industry. These guides can be designed by food industry
and are then subject to approval of competent authorities. They are a recognised tool to

Table 1. Legal framework applying to the local, direct trade of meat and meat products from wild game.
Product Subtypes Requirements as in the act on direct marketing1 Other requirements2
Fresh meat from carcasses, skin-on, • marketed by the hunter • Regulation (EC) No.178/2002
wild game cooled • has undergone ante and post mortem inspection3 • national guide to personal hygiene (based on Regulation
• evisceration and cooling to max. 7 °C or 4 °C for large and small (EC) No. 852/2004, Annex II, Chapter VIII)4
game, respectively, without undue delay • national Act on animal-by-products5
• transportation and storage must not cause contamination of
meat and temperatures must be maintained
• must be marketed with 7 days post mortem

meat cuts, cooled as above, and in addition: as above, and
or deep-frozen; also • meat must be labelled (‘Wildfleisch aus Direktvermarktung’ plus • Regulation (EC) No. 852/2004 (Annex II)
packaged; (frozen indication of region of origin) • national legislation on labelling of foodstuffs 6
carcasses) • Austrian Food Codex7
Meat products Act on direct marketing does apply only implicitly, but an act on as above, and
local trade of foods8 sets into force parts of Regulation (EC) No. • muscle tissue must comply with requirements as above
853/2004. • all other ingredients must be food grade
1 108. Verordnung der Bundesministerin für Gesundheit und Frauen über die Direktvermarktung von Lebensmitteln (Lebensmittel-Direktvermarktungsverordnung), BGBl.
II, Nr. 108/2006, as amended; the underlying Austrian Food Safety Act applies also.
2 Not exhaustive.
3 Also Trichinella-inspection, as applicable.
4 ‘Leitlinie zur Sicherung der gesundheitlichen Anforderungen an Personen beim Umgang mit Lebensmitteln’. Available at: http://www.bmgfj.gv.at/cms/site/attachments/5/9/5/
CH0819/CMS1073644716719/gesundheits-anforderungen1.pdf.
5 Verordnung der Bundesministerin für Gesundheit, Familie und Jugend über nähere Bestimmungen zum Umgang mit tierischen Nebenprodukten (Tiermaterialien-
Verordnung), BGBl. II Nr. 484/2008.

6 Verordnung über die Kennzeichnung von verpackten Lebensmitteln und Verzehrprodukten, BGBl.Nr. 72/1993, as amended.
7 Codex Alimentarius Austriacus, 4th Ed., chapter B14. Available at: http://www.bmg.gv.at/cms/site/attachments/4/9/6/CH0832/CMS1167207128242/b_14_fleisch_und_
fleischerzeugnisse.pdf.
8 92. Verordnung der Bundesministerin für Gesundheit und Frauen über Lebensmittelhygieneanforderungen an Einzelhandelsunternehmen (Lebensmittel-
Einzelhandelsverordnung), BGBl. II, Nr. 92/2006, as amended.

The lower Austrian model
261
establish GHP, and also for HACCP (Hazard Analysis and Critical Control Points) based
food safety assurance systems. GHP and HACCP implementation guides have been issued in
several EU countries. Recently released textbooks in Austria on inspection and on processing
of wild game were designed to address all relevant issues laid down in EU food hygiene
legislation (Winkelmayer et al., 2007, 2008). Particular emphasis is put on the requirements
for food premises which are temporary or used primarily as a private dwelling house (in the
sense of Regulation (EC) No. 852/2004, Annex II, Chapter III (EC, 2004a)). The minimum
documentation recommended for hunters operating a temporary food premise is: (1) a signed
copy of the guide for personal hygiene; (2) a master data sheet; and (3) a two-pages checklist
for each workday which includes personal hygiene, condition and cleanliness of the premise
and food-contact surfaces, condition of cooling facilities and maintainance of the required
temperatures, results of the inspection of the game carcasses, water quality and disposal of
waste/by-products (Winkelmayer et al., 2007). Hunters operating permanent food premises
are offered separate checklists addressing individually the chapters in Annex II of Regulation
(EC) No. 852/2004 (EC, 2004a). These checklists are based a system already operational for
meat industry (slaughterhouses, butchers, etc.) in Austria.
3.2 Education and training
Food safety is the primary responsibility of the producer. In order to enable the producer to
fulfill this obligation, training courses for direct marketing of fresh meat and meat products
from wild game are offered in Lower Austria (Table 2). These courses have a modular structure
and rely on a basic training in game inspection and hygiene (Winkelmayer et al., 2011). For
the practicals, a network of agricultural schools (equipped with approved cutting rooms,
cooling rooms) provides a reliable infrastructure.
3.3 Survey on structure and product range
From December 2008 – April 2009, a survey was conducted on the direct markting of meat
and meat products from game in Lower Austria (Fettinger and Paulsen, 2009). All hunters
which had undergone training courses on direct marketing, or which had registered as
direct suppliers of game meat, received a questionnaire, to collect basic information of
infrastructure, hygiene level and products (fresh meat or meat products), see Table 3. These
ca. 420 persons represent ca. 1.4% of licensed hunters in Lower Austria (Pontasch, 2008). All
participants submitting a completed questionnaire were entitled to send up to 15 samples
(fresh meat or products from wild game) for microbiological examination in the year 2009
(Fettinger et al., 2010).
A total of 109 completed questionnaires (ca. 26%) have been returned. 77% of the hunters
operated temporary game processing premises, and 23% had permanent facilities. Selected
findings are presented in Table 3. Such data can be particularly useful when the Competent
Authority has to establish risk-based inspection schemes.

Table 2. Training courses for local, direct trade of meat and meat products from wild game.
Theoretical lessons lectured by official Practicals under supervision of a specialised

veterinarians butcher and an official veterinarian
Fresh meat • legislation • skinning of carcasses

(ungulates) • required infrastructure • preparation of boneless cuts
• hygiene during meat processing • vacuum-packaging
• humans and animals as source of foodborne • declaration
disease • assortment of meat cuts
• factors affecting food safety and shelf life,
• documentation
Meat products as above, plus as above, plus
• Austrian food codex • production of 2-3 different pasteurised
• food declaration incl. QUID cured meat products (e.g. frankfurter-type
• good manufacturing practice in meat sausage and paté)
products • food labelling
• meat technology (ingredients, specific
machinery for meat products; mincing,
curing, smoking and heating)
• temperature control as a part of HACCP
based food safety
• documentation
Small game see ‘fresh meat’
3.4 Microbiological own-checks
Microbiological self-checks can be a useful tool to document compliance with GHP and to
assess product quality and safety. The prerequisite is, of course, that there are standardised
procedures for evisceration, cooling, storage, cutting, etc. (Fettinger et al., 2010).
Based on the fact that meat from game can be marketed in the same way as that from farm
animals, and that both types of meat can be displayed in the same shelf in butchers shops,
it is reasonable to suggest the same microbiological limits for game meat as for meat from
farm animals, provided that GHP procedures are strictly adhered to. This assumption has
been substantiated in various studies (e.g. Paulsen et al., 2003; Paulsen and Winkelmayer,
2004; El-Ghareeb et al., 2009).
To check this hypothesis, submitted game meat and meat product samples were categorised
(Fettinger et al., 2010) and then examined according to a product-specific catalogue consisting
of food safety criteria as defined in Regulation (EC) No. 2073/2005 (EC, 2005) and other
criteria laid down in specifications of fresh meat from slaughter animals (AMA, 2004) and a
collection of microbiological limits for meat products (Eisgruber and Bülte, 2006).

Table 3. Results of a survey on the structure of local marketing of meat from wild game in Lower Austria
(data in %), 2008-2009 (after Fettinger et al., 2010).
Specific education of hunters* specific training courses 83

professional education (butcher, etc.) 17
self-study of textbooks, etc. 36
Assistance of none 42
butchers or other professionals 38
non-professionals 20
Premises permanent 23
temporary 77
Cooling facilities* own cool room for game carcasses 59
shared cool rooms 41
additional refrigerator for meat cuts and meat products 47
Production of* fresh meat 100
meat products 43
Game species* roe deer 97
wild boar 70
red deer 41
brown hare 43
pheasant 35
Meat traded* refrigerated 93
deep frozen 54
vacuum-packed 71
Meat supplied to* self-supply 86
final consumer 79
local butcher 8
restaurants 32
* multiple answers possible
To date, 13 of 109 interested hunters have submitted a total of 55 samples, 22 of which were
fresh meat. The pathogens Salmonella spp. and Listeria monocytogenes were not detected in
any sample. From interviews conducted with a number of participants, this overall moderate
acceptance was most probably caused by the discontinuous (seasonal) and low-scale mode
of production.
4. Conclusions
Although a training and product evaluation system for hunters supplying game meat and
meat products directly to the final consumer or to local food retail establishments has been
established, it has become clear that traditional training must be augmented by new ways, e.g.
by contests for game meat (products) in the framework of local conferences. The submission
of products for such contests would require that certain chemical and microbiological

specifications are met and that evidence is presented that GHP is adhered to. In turn, such
product contests would communicate to the public that the hunters are able to supply game
meat and meat products of consistent quality under observation of hygiene rules. Such a pilot
conference has already been held in November 2009.
Also, the possibility to rent the infrastructure of the network of agricultural schools spread
over entire Lower Austria should be improved. This would allow hunters which are not willing
or able to do investments in own infrastructure to work and process meat under professional
conditions.
Acknowledgements
This contribution is based on presentations given at the conference ‘Hygiene of wild game meat
and its safety within the food chain’, Prague, 22-24 April 2009, and on the 50. Arbeitstagung
des Arbeitsgebietes ‘Lebensmittelhygiene’ der DVG, Garmisch-Partenkirchen, 29 September
- 2 October 2009.
References
AMA (Agrar Markt Austria), 2004. Microbiological criteria for meat. Available at: www.ama.gv.at. Accessed 6
February 2010.
Bauer, F., Vali, S. and Paulsen, P., 2007. Formation of biogenic amines and fat oxidation products in wild boar
salami. Euro. Food Chem. XIV, 29-31.08.2007, Paris, France, pp. 285-288.
EC (European Commission), 2002. Regulation (EC) No. 178/2002, laying down the general principles and
matters of food safety. O.J. L 31/1.
of 29th of April 2004 containing specific rules for the hygiene of foodstuffs. O.J. L 139/55.
EC (European Commission), 2005. Regulation (EC) 2073/2005 of the European Parliament and the Council of
15th of November 2005 on microbiological criteria for foodstuffs. O.J. L 338.
Eisgruber, H. and Bülte, M., 2006. Mikrobiologische Kriterien und Mykotoxin-Höchstgehalte für Lebensmittel.
Behrs Verlag, Hamburg, Germany.
condition and shelf life of meat from hunted game birds. Eur. J. Wildl. Res. 55, 317-323.
Fettinger, V. and Paulsen, P., 2009. Direct marketing of wild game: microbiological own-checks (in German).
Österreichs Weidwerk 9, 10-13.
Fettinger, V., Smulders, F.J.M., Winkelmayer, R. and Paulsen, P., 2010. Lower Austrian model for microbiological
own-checks in the local game meat chain (in German). Rundsch. Fleischhyg. Lebensmittelüberw. 62, 118-120.
Kainz, R. and Paulsen, P., 2005. Neue EU Hygienevorschriften für Fleisch von jagdlich erlegten Wildtieren. Wien.
Tierärztl. Mschr. 92, 150-156.
Paulsen, P. and Winkelmayer, R., 2004. Short Communication: Seasonal variations in the bacterial contamination
of game carcasses in Austria. Eur. J.Wildl. Res. 50, 157-159.

Paulsen, P., Hilbert, F., Winkelmayer, R., Mayrhofer, S., Hofbauer, P. and Smulders, F.J.M., 2003. Zur tierärztlichen
Fleischuntersuchung von Wild, dargestellt an der Untersuchung von Rehen in Wildfleischbearbeitungsbetrieben.
Arch. Lebensmittelhyg. 54, 137-140.
Pontasch, W., 2008. Jagd-Almanach 2009. Österr. Jagd- und Fischereiverlag, Vienna, Austria.
Winkelmayer, R., 2006. Food safety inspection and monitoring of meat from wild game in Austria. Lecture held
at the Assembly General Meeting of the ECVPH, 7.-8. December, Lyon, France.
Winkelmayer, R., Malleczek, D., Paulsen, P. and Vodnansky, M., 2005. Röntgenanatomische Untersuchungen
beim Rehwild in Hinblick auf den optimalen Zielpunkt für den tierschutzgerechten und wildbrethygienisch
einwandfreien Schuss. Vet. Med. Austria 92, 40-45.
Winkelmayer, R., Paulsen, P., Lebersorger, P. and Zedka, H.F., 2008. Wildbret-Hygiene, 3rd ed., Zentralstelle Österr.
Landesjagdverbände, Vienna, Austria, 222 pp.
Winkelmayer, R., Paulsen, P., Lebersorger, P. and Zedka, H.F., 2007. Wildbret-Direktvermarktung. Zentralstelle
Österr. Landesjagdverbände, Vienna, Austria, 176 pp.
Winkelmayer, R., Stangl, P.V., Paulsen, P., 2011. Assurance of food safety along the game meat production chain:
Inspection of meat from wild game and education of official veterinarians and trained persons in Austria.
In: Paulsen, P., Bauer, A., Vodnansky, M., Winkelmayer, R. and Smulders, F.J.M. (eds.). Game meat hygiene
in focus: microbiology, epidemiology, risk analysis and quality assurance. Wageningen Academic Publishers,
Wageningen, the Netherlands, 245-258

Approaches to game hygiene in the province
Belluno (Italy): from training to meat microbiology
Carlo V. Citterio1, Patrizia Bragagna2, Enrico Novelli3 and Valerio Giaccone3
1IstitutoZooprofilattico Sperimentale delle Venezie, SCT2 Belluno, Via Cappellari 44/A,
32100 Belluno, Italy; ccitterio@izsvenezie.it
2UlSS 1, Belluno, Via S. Andrea 8, 32100 Belluno, Italy
3Dipartimento di Sanità Pubblica, Patologia Comparata e Igiene Veterinaria, Università degli
Studi di Padova, Viale dell’Università 16, 35020 Legnaro (PD), Italy
Summary
In the last decades, the number of animals shot in the Italian Alps during the regular hunting
seasons has dramatically increased, which is particularly the case for wild ungulates. In
these areas game is considered an important source of meat, and an increasing interest of
consumers for game meat with high added value and favourable nutritional properties is
noticeable (Hoffman and Wiklund, 2006). Nevertheless, targeted measures to assure game
meat hygiene are still lacking in Italy, and much work is needed to convince hunters and
consumers in general of the importance of this topic. This contribution summarises various
complementary approaches to game meat hygiene, as initiated in the territory of the province
Belluno, in the Italian Eastern Alps, i.e. the centre of the Dolomites, where red deer, roe deer,
chamois, mouflon and wild boar are hunted.
Keywords: hygiene training, meat microbiology
1. Passive sanitary surveillance
Sanitary surveillance, together with a better management of wildlife and livestock, is one of
the first actions to ensure quality and safety of game meat.
To be effective, sanitary surveillance should involve different stakeholders in the territory

– including hunting associations, authorities for animal and human health and for wildlife
management, laboratories and research institutes – with the aim to collectively elaborate a
surveillance system. In the province Belluno, a ‘passive surveillance programme’ has been
carried out in 2009, by developing a specific vademecum and cards for data collection, as
shown in Figure 1 and 2.
This activity also included a 2-3 days training for hunters and gamekeepers, consisting of
basic instructions on:
1. game anatomy and physiology;
2. game pathology;
3. good practices in game meat handling and conservation;
4. main alterations of game meat;

Figure 1. Vademecum for passive sanitary surveillance in hunted game.
Figure 2. Cards for data collection by passive sanitary surveillance.

Approaches to game hygiene in the province Belluno (Italy)
5. eco-epidemiology of disease in game;

6. regulations about game and game meat;
7. terminal ballistics (e.g. Figure 3).
For topics 1-3, a practical session was also held.
2. Game meat microbiology

From Belluno, data about game meat microbiology were also available (Bragagna et al., 2004).
Microbiological analyses were performed on 90 muscle samples from various wild ungulate
species (39 roe deer, 27 red deer, 11 chamois, 8 mouflon and 5 fallow deer). The mean values
(log10 cfu/g) of microbiological counts of core tissue samples are summarised in Table 1.
Pathogens such as Salmonella spp. or Listeria monocytogenes were not isolated. The Total Viable
Count mainly consisted of Gram negative bacteria, such as faecal coliforms. A substantial
difference in microbial carcass contamination was found between carcases correctly stored
(i.e. in cold rooms at 0-4 °C) and carcases stored in less refrigerated places, such as cellars,
where generally temperatures of 7-14 °C prevail depending on the season. In the latter case,
levels of spoilage bacteria were definitely higher. Similar observations have been reported by
Paulsen and Winkelmayer (2004).
Figure 3. Example of a practical session in terminal ballistics.

Table 1. Microbiological profile (mean values) of muscle core samples taken from 90 specimens of wild
ruminants in Belluno province, Italy.
log10 cfu/g
Total viable count 4.52

Total coliforms 4.32
Faecal coliforms 3.46
Sulphite reducing clostridia 2.11
Micrococcus spp. 3.16
Staphylococcus spp. 2.15
Lactobacillus spp. 4.55
Pseudomonas spp. 3.59
Aeromonas spp. 1.90
Acknowledgements
The authors are grateful to drs. P. Catellani, S. Balzan, P. Capovilla and to the Belluno
Provincial Office for Hunting And Fishing and to F. Obber and V. Boscolo for the editing of
the manuscript.
References
Bragagna, P., Catellani, P., Balzan, S. and Sommavilla, G., 2004. Microbiological analysis on samples of hunted
game meat. Proceedings of the IV Conference of Associazione Italiana Veterinari Igienisti (AIVI),Vicoforte
(CN, Italy), pp. 381-386.
Hoffman, L.C. and Wiklund, E., 2006. Game and venison – meat for the modern consumer. Meat Science 74,
197-208.
Paulsen, P. and Winkelmayer, R., 2004. Seasonal variation in the microbial contamination of game carcasses in
an Austrian hunting area. Eur. J. Wildl. Res. 50, 157-159.

Section 4
Muscle biology and meat quality
Key contributions
The muscle biological background of meat quality
including that of game species
and Veterinary Public Health, University of Veterinary Medicine, Veterinärplatz 1, 1210
Vienna, Austria; peter.hofbauer@vetmeduni.ac.at
Summary
A review of the major biological mechanisms determining the physical-chemical and sensory
quality traits of whole tissue meat (colour and water-holding of fresh- and tenderness and
flavour of cooked meat) is presented. The effect of various ante mortem and processing factors,
affecting sensory quality traits of the various major animal meat species, are summarised
and reference is made to effects reported for game meat species.
Keywords: muscle biology, meat quality, colour, water-holding, tenderness, flavour
1. Introduction
The concept of ‘meat quality’ includes many aspects such as hygiene and food physiological,
technological and sensory properties (Hofmann, 1990). Whilst consumers can assume that
the meat industry generally adheres to Good Hygiene and Manufacturing Practices which
guarantee the meat safety- and allow storage of meat during a certain period, they usually
are less aware of the many technologies that are primarily directed at improving sensory
traits of meat.
The quality of fresh meat, as perceived when buying, preparing and consuming whole tissue
cuts, is determined by ante and post mortem factors having interfered with muscle biological
events in skeletal muscle. Many of these are common to all animal species and the effects of
various processing technologies affecting meat quality (for instance, that of beef, pork, and
poultry) have over the past decades been studied extensively. With few exceptions, game
meat species have received less attention and the available information is still scarce and
fragmented.
The purpose of the present contribution is first and foremost to review the major muscle
biological mechanisms and processing effects underlying the sensory quality of meat in
general. In addition, some data on game meat quality recently generated in our laboratory
have been included.

2. Muscle structure, composition and function
Skeletal muscle (expressed as percentage wet weight) is composed of 75% water, 19% protein,
soluble organic compounds (3.5%), variable amounts of lipids (0.5-3%), carbohydrates (1-2%)
and small amounts of minerals and vitamins (Lawrie and Ledward, 2006).
However, the visual appearance and the physiological and biochemical characteristics of
skeletal muscles vary considerably, reflecting the proportion of fibre types present in the muscle
cells. The commonly used fibre type classification is based on the contraction speed (fast or
slow) and the energy metabolism (oxidative or glycolytic) of the fibres. The biochemical traits
associated with different fibre types (e.g. ATP concentration, calcium-, myoglobin-, glycogen-,
lipid-, proteinase and their inhibitors content, and enzyme activities) reflect the diversity
of muscle fibres and are related to their physiological function exerted in the living animal
(Pearson and Young, 1989). In this respect, meat quality parameters such as colour, flavour,
juiciness and tenderness are fibre type dependent (for a review: see Monin and Ouali, 1991).
The coherent structure of muscle fibre bundles is maintained by the intramuscular connective
tissue. Three connective tissue structures are distinguished in muscle, i.e. the epimysium
(a fibrous sheath of connective tissue which surrounds the entire muscle), the perimysium
(a three dimensional collagen network surrounding the muscle fibre bundles) and the
endomysium (a layer of fine connective tissue fibres encircling individual muscle fibres).
Not more than 5% of the total water of muscle is directly bound to hydrophilic groups of the
proteins, the rest is divided between the so-called ‘free water’ (i.e. immobilised by the physical
configuration of the proteins but not bound to them) and the so-called ‘loose water’, which
is expressed when the water-holding capacity drops (Hamm, 1972).
Fat content varies in quantity and composition between muscles and species, males generally
having less than females. Many intracellular lipids are associated with membrane structures.
In addition, considerable amounts of lipids are present in the perimysium surrounding
myofibre bundles. Macroscopically this is perceived as ‘marbling’.
Muscle proteins may be classified as sarcoplasmic, myofibrillar and stroma proteins.

Sarcoplasmic proteins are soluble in water or salts of low ionic strength. Many of them are
enzymes involved in the breakdown of glycogen. Myofibrillar proteins, which constitute
the major part of the muscle’s contractile mechanism, are only soluble in solvents with
higher ionic strength than required for the extraction of sarcoplasmic proteins. The main
myofibrillar proteins are myosin, actin, and the troponin-tropomyosin complex. Stroma
proteins include collagen, elastin and reticulin, all present in connective tissue, and the
proteins which are found in the membrane systems of the muscle cell organelles such as the
mitochondria and sarcoplasmic reticulum.
The myofibrillar proteins are arranged in repeating units of actin and myosin. The thin
myofilament is composed of two strands of (‘filamentous’) F-actin, which consist of
polymerised (‘globular’) G-actin. Two strands of F-actin are coiled around each other and
have a very precise length. The actin helix is associated with tropomyosin, a double helix of

The muscle biological background of meat quality including that of game species
two unidentical peptide chains, and the troponin complex (i.e. consisting of three polypeptide
subunits C (binding calcium ions and thus effecting a change of conformation), I (inhibiting
the interaction of actin to myosin) and T (the tropomyosin-binding subunit), which cements
the tropomyosin to the actin helix). Figure 1A schematically presents this arrangement.
The troponin complex is thought to turn on the contraction process by binding Ca 2+ ions that
are released from the sarcoplasmic reticulum after a nervous stimulus. The thick myofilament
consists of packed myosin molecules (Figure 1B) each of which has a helical tail and a head
consisting of two globular units. The myosin heads contain a site with ATPase activity and
a site which forms cross bridges that interact with actin to form actomyosin during muscle
contraction.
Figure 2 shows a diagrammatic representation of the main (ultra)structural features of muscle.

The I-band mainly consists of actin filaments that are attached to a transverse structure
called Z-lines. The spacing between Z-lines is referred to as sarcomere length. Myosin fills
the entire length of the A-band. A clear central H-zone contains the M-line representing the
cross bridges between separate myosin filaments.
In developed striated muscle various ‘cytoskeletal’ proteins serve the role of maintaining cell
shape and integrity and they are important for the function of movement. An example is
titin, which is referred to as the ‘gap-filament’, anchored inside the myosin as a core protein
and, running through the Z-lines, interconnects myosin filaments of adjacent sarcomeres.
Figure 1. (A) The association of actin with the troponin-tropomyosin complex. (B) The structure of the
myosin molecule (Cohen, 1975).

epimysium
perimysium connective tissue
endomysium
muscle muscle bundle
N-line
H-zone
Z-line titin M-line Z-line

sarcomere
Figure 2. A schematic overview of the structure of skeletal muscle (Geesink, 1993).
3. Post mortem muscle physiology; the conversion of muscle to meat
Figure 3 is a diagrammatic representation of the main muscle physiological events, which are
related to sensory meat quality traits.
In the living animal the blood circulation provides the muscle fibres with oxygen and
glucose. Whenever muscle action is required, nervous stimuli effectuate a depolarisation of
the sarcoplasmic reticulum membrane. This results in a release of Ca2+ ions which activate
the enzymes that convert glycogen into pyruvate and eventually into carbon dioxide and
water in the mitochondria (Krebs cycle). In the course of this process adenosine diphosphate
(ADP) is phoshorylated to adenosine triphosphate (ATP). Initially the muscle relies on the

Figure 3. Diagrammatic presentation of the physiological events effecting the conversion of muscle to
meat (After Smulders, 2007).
muscle specific creatin phosphate (CP) to supply the high energy phosphate. The conversion
of ATP to ADP directly supplies the energy needed for muscle contraction and metabolic
activities. This is affected by Ca2+ ions released by the sarcoplasmic reticulum, which stimulate
myosin ATPase. Muscle relaxation also requires ATP, not only because the calcium pump
transporting Ca 2+ back into the sarcoplasmic reticulum is energy requiring, but also because
myosin rods will only be allowed to shift out of the sheaths formed by the actin filaments
provided ATP is present in its function as ‘plasticizer’. Under anaerobic conditions ATP is
resynthesized via the comparatively inefficient glycolysis leading to the formation of lactate,
which may be resynthesised to glycogen in the liver via the Cori cycle (gluconeogenesis).

When an animal is slaughtered it is subjected to a state of shock which acts upon the muscle as
a complex of nervous stimuli. Since carbohydrate and oxygen supply has ceased, the muscles
are dependent on glycolysis for their energy synthesis from the moment the muscle specific
creatin phosphate (CP) reserves have been depleted. Consequently comparatively little ATP is
resynthesised. Moreover, the enzymes catalysing glycolysis and ATP breakdown are activated
and lactate and metabolites such as adenosine monophosphate (AMP) are formed. As a result
the muscle pH gradually falls. Rigor mortis sets in when too little ATP is available to keep
the actin and myosin filaments apart in a relaxed state. This occurs when the ATP residue is
approximately 20% of its initial concentration (Hamm, 1977).
The biochemical reactions in the period before onset of rigor mortis – generally referred to
as the period of ‘conditioning’ – as well as those resulting from the subsequently occurring
post mortem muscle proteolysis – also known as ‘aging’ – have a great impact on sensory
meat quality characteristics.
4. Major sensory characteristics of meat
Whilst major interrelated phenomena occurring post mortem and valid across the various
meat species have been included in Figure 3, the following section discusses in more detail
the relevance of the various factors that determine the major quality traits of meat. Wherever
meaningful, particular reference will be made to those factors that might have relevance to
the quality of game meat. The reader is referred to another contribution to this Volume for
an overview of methods to quantitatively assess these various meat quality characteristics
(Hofbauer and Smulders, 2011).
4.1 Colour of fresh meat
The colour of fresh meat is determined both by light absorption (dependent on the contents
of pigments (i.e. apart from the cell pigment cytochrome, particularly hemoglobin and
myoglobin)) and light being reflected from water present on the meat surface (largely related
to the water-holding properties of muscle proteins and the density of the myofibrillar matrix;
see below). Provided an animal is properly exsanguinated, the role of hemoglobin is relatively
minor: fresh meat contains no more than 0.3% residual blood.
4.1.1 Myoglobin – related factors
The pigment primarily responsible for colour is myoglobin (Mb). Like hemoglobin in blood,
its physiological function is to store oxygen and deliver it to the muscle whenever necessary.
Upon binding of oxygen, its colour changes from purple to an attractive cherry-red which
reflects the presence of oxymyoglobin (MbO). When the heme-iron oxidises (ferrous iron,
Fe2+, turns into ferric iron Fe3+), metmyoglobin (MMb) is formed which has lost the ability
to bind oxygen. Concurrently colour turns into an unattractive greyish-brown. Only through
reduction with the aid of reducing enzymes can MMb be converted back into physiologically
active (oxygen-binding) Mb.

The relationship between partial pressure of oxygen (pO2) and the chemical form of Mb
(Mb, MbO or MMb) is presented in Figure 4. Depending on the presence of oxygen, be it
at atmospheric- or at higher pressures as for instance prevailing in modified atmosphere
packaging, the surface of any piece of meat will have a more or less thick superficial layer of
MbO, which is being replaced by MMb at depths where oxygen penetration is insufficient,
and finally by Mb in the core of the muscle where pO2 is zero.
When interacting with hydrogen peroxide (H2O2) or hydrogen suphide (H2S), Mb transforms
into the green pigment cholemyoglobin (ChMb) or sulphmyoglobin (SMb). The formation of
one or both of these pigments is responsible for the green discoloration of meats with severe
bacterial contamination.
4.1.2 Fresh meat colour as affected by its water-holding properties
The ability of a muscle matrix to hold on to water has a large impact on meat colour. The lower
the water-holding capacity of meat (see below) the more water molecules will be released
onto- and the more light will be reflected from its surface.
The well-known Pale-Soft-Exudative (PSE) condition predominantly found in pork is largely

the result of sarcoplasmic protein and myosin denaturation, caused by a combination of
high carcass temperatures associated with stress (malignant hyperthermia- or porcine stress
syndrome) and the resulting extremely fast pH decline. As a consequence, PSE meat binds
water very poorly and the meat surface reflects light causing its appearance to be pale.
Certain stressful ante mortem conditions, e.g. those associated with physical exhaustion as
seen after intensive hunting, can cause healthy animals to develop so-called Dark-Firm-Dry
(DFD) meat, which is characterised by a high ultimate pH, a purplish-black colour, a firm
100%
MbO
% total pigment
MMb
Mb
0 20 40 60 80 100 120 140 160

pO2 (mm Hg)
Figure 4. The effect of partial pressure of oxygen on the prevalence of various myoglobin forms (Smulders,
2007).

texture and a dry sticky surface. The condition is predominantly reported to occur in beef
and pork, but is similarly relevant for game species such as deer (Wiklund and Smulders,
2011) subjected to stressful conditions, e.g. regrouping, being chased or being subjected to
inappropriate transport conditions. The depletion of muscle glycogen stores caused by these
activities results in a lower than normal muscle acidification (higher ultimate pH values), and
consequently in unusually high electrostatic binding of water by the muscle proteins (i.e. an
extremely low release of water to its surface (see Section 4.3.1)) and hence less light reflectance
(explaining its darker appearance).
It is questionable if the PSE or DFD condition occurs in other species than those mentioned,
although there are reports suggesting their prevalence for instance in poultry (e.g. Van Hoof
and DeZeure-Wallays, 1980).
4.1.3 Colour stability of meat
In essence, the stability of the colour of fresh meat is characterised by the muscle’s ability
to keep Mb in the oxygenated form and to prevent the formation of MMb. Although none
of them react independently of the other, several biological factors impact on meat colour
stability. Major ones recognised by Faustman and Cassens (1991) are: (1) muscle pH, low
values promoting the formation of MMb, (2) temperature, higher muscle temperatures
favouring increased formation of MMb and dissociation of oxygen from MbO, (3) relative
humidity (rh), lower rh’s leading to more desiccation and a darker surface colour, (4) exposure
to light (particularly the ultraviolet part) resulting in more MMb formation, (5) bacterial
contamination, the exponential growth phase of the spoilage flora coinciding with the highest
oxidation, (6) lipid oxidation, (7) partial pressure of oxygen (pO2), where zero (not low) pO2
prevents MMb formation and only pO2’s around or above 80 mm Hg promote the desirable
MbO formation (see Figure 4), (8) the presence of MMb reducing enzyme systems, prolonged
refrigerated storage leading to a considerable loss of reducing activity (‘fading’ of meat), (9)
pre-slaughter stress, leading to an aberrantly fast pH decline or high ultimate pH values (see
above), and finally (10) muscle dependent sensitivity to discoloration, probably related to their
oxidative capacity (fibre type).
4.1.4 Ante mortem and processing effects on meat colour – summarised
Factors affecting the ultimate appearance (colour) of fresh meat (Smulders et al., 1991) include:
(1) species effects or genetic variation within species (responsible for differences in oxygen
consumption rate of mitochondria which counteract MbO formation, or determining the
vulnerabilty to stress and the associated PSE condition); (2) sex and age (e.g. as reflected in
the darker colour of bull vs. cow meat); (3) plane and quality of nutrition (e.g. determining
the degree of marbling and iron content); (4) inappropriate ante mortem handling of animals
leading to PSE or DFD (see above); (5) accelerated carcass processing methods such as electrical
stimulation (which, when not applied correctly, can lead to increased protein denaturation
(e.g. Eikelenboom and Smulders, 1986)) or hot boning allowing faster cooling of primals
and reduced protein denaturation (Van Laack, 1989); and, finally (6) modified atmosphere
packaging (allowing an increased formation of MbO, provided high oxygen concentrations
are present in the gas mixture).

Figure 5. Fragmentation of myofibrils adjacent to Z-lines (see arrows) in bovine longissimus muscle after
14 days of storage at 2-4 °C (by courtesy of P. Koolmees).
4.2 The tenderness of meat
The main components thought to determine the tenderness of meat are connective tissue,
myofibrillar proteins matrix and fat. As the role of fat is primarily that of a ‘dilutant’ of the
muscle matrix (a high degree of marbling generally being associated with better tenderness
and only extremely low contents <1% being reported to promote toughness) the following
sections concentrate on the contribution to tenderness of the former two components.
4.2.1 The connective tissue component of tenderness
Connective tissue serves an important physiological function in muscle. It supports muscle

fibres and carries over the forces generated by myofibrils to the skeleton. Connective tissue
predominantly consists of collagen (65-95%). In addition, the less rigid elastin prevails in
lower amounts (5-35% of the total connective tissue, dependent on the muscle), particularly
in the epimysium.
Not only the collagen content, but also its type and the degree of cross-linking between the
constituting tropocollagen molecules affect the tenderness of meat. Various isoforms of
collagen exist. In muscles, types 1-5 are important, each type having distinct properties, For
instance, type 3 is markedly more heat stable than is type 1 (McCormick, 1994). Consequently,

the distribution of collagen isoforms largely determines the solubility of connective tissue
(Stanton and Light, 1987). Contrary to the myofibrillar component of tenderness, which is
significantly affected during post mortem storage of raw meat, changes in connective tissue
are only observed in the course of heating.
Although the ratio connective tissue: muscle substance decreases during growth, the degree of
cross linking increases, which explains the relative toughness of meat from older vs. younger
animals. It was once thought that connective tissue was the overriding factor explaining
tenderness differences between meats. However, research in the past decades has clearly
shown that its role is restricted to its determining the so-called ‘background toughness’,
which obviously is muscle- but also species dependent and is particularly related to the
content of collagen type 3 (Dransfield, 1977, Light et al., 1985). Whereas in the course of
storage the properties of collagen barely change, heating during meat preparation will affect
its solubility markedly, provided temperatures >80 °C are reached at which collagen gelatinises
and becomes soft.
4.2.2 The myofibrillar component of tenderness
Density of the myofibrillar matrix

Although a clear relationship between sarcomere length and tenderness score of meat does
not always exist, there appears to be general consensus that long sarcomeres are associated
with tenderer-, short sarcomeres with tougher meat (unless ‘super-contraction’ has occurred
beyond 40% of the muscle’s rest length of about 2 µm (Marsh and Carse, 1977)). However,
said relationship is not linear and appears to be only valid for muscles with slow glycolysis
(Smulders et al., 1990). Preventing muscles to contract during the onset of rigor mortis (as
achieved through the ‘tenderstretch’ process, e.g. by pelvic suspension), also leads to a tenderer
end-product (Smulders et al., 1992).
The degree of muscle contraction during rigor onset is markely affected by the muscle’s
temperature decline. Although at low temperatures ATP breakdown is slower (as the
glycolytic enzymes’ activity is lower during refrigeration), at critically low temperatures <6 °C
mitochondrial release of Ca2+ increases while at the same time the calcium pump responsible
for pumping Ca2+ ions back into the sarcoplasmic reticulum fails to function properly. The
resulting increased concentration of sarcoplasmic Ca2+ leads to an intensive contraction known
as cold shortening (Conforth et al., 1980). Cold shortening occurs primarily in meat from
animal species exhibiting slow muscle glycolysis (e.g. ruminants). Beef processors using fast
chilling regimes therefore generally rely on electrical stimulation which accelerates glycolysis
and thus prevents low temperatures being reached when muscle pH is still high (Smulders and
Van Laack, 1995). In this context, Bendall (1973) reported that combinations of muscle pH
>6.2 while temperatures are already below 12 °C should be considered particularly hazardous.
A phenomenon known as ‘heat-shortening’ or ‘rigor contracture’ may become relevant when

rigor sets in at temperatures >25 °C, e.g. when muscle is not chilled immediately after the
animal’s death (as is often the case in game animals left unrefrigerated for too long). This
situation also explains the sometimes extremely tough meat found in PSE pork.

Myofibrillar proteolysis
In post mortem tenderisation, which results from a loss of the muscle’s structural integrity
(see Figure 5), proteolytic enzymes play a dominant role. Research has concentrated on two
enzyme systems, viz. the calpains (i.e. calcium activated sarcoplasmic enzymes active at high
muscle pH) and the cathepsins (lysosomal enzymes active at low pH); (for more extended
reviews: see Roncalés et al. (1995), Smulders et al. (1999), Ouali (2007)).
Calpains belong to the cystein group of proteases, which have an optimum pH between 7
and 7.5 and need calcium to become active. The efficacy of the calpains’ proteolytic action
is based both on their breaking down of structural myofibrillar elements, and on their own
autolysis. The proteolytic rate of both these processes depends heavily on pH and temperature
(Dransfield, 1994). At least 3 components of the calpain system have been distinguished,
i.e. the proteinases µ-calpain (active at micromolar concentrations of Ca 2+) and m-calpain
(active at millimolar concentrations of Ca2+) and the specific inhibitor calpastatin. Binding
of calcium induces autolysis, by which the enzyme is both activated and ultimately broken
down (Figure 6). The m-calpains concentration remains fairly constant throughout ageing,
indicating its contribution to meat tenderisation is minimal (Koohmaraie et al., 1987).
Inactive Inactive
Autolysis
Active Inactive
Autolysis
Active Inactive
Autolysis Ca2+
Inactive Calpain
Calpastatin
Figure 6. Schematic diagram showing the interaction of calpain with Ca2+ and calpastatin, and autolysis
of calpain (Geesink, 1993).

Cathepsins are lysosomal endopeptidases, 8 of which have been found in skeletal muscle
(cathepsin A, B, C, D, H, L, J and carboxypeptidase B). Cathepsin B, D, H and L appear to
have the highest activity in skeletal muscle and are active at pH values ranging from 2.5-
6.0. Cathepsins activity is regulated by a series of inhibitors, the cystatins, prevalent in the
sarcoplasm. It has been reported that at least cathepsins D, H and L may break down actin
and myosin, albeit only provided temperatures are higher than 2-4 °C. Hence, it is assumed
proteolysis by cathepsins is probably only relevant when meat is held at elevated temperatures
(Goll et al., 1989).
4.2.3 Ante mortem and processing effects on meat tenderness – summarised
Factors affecting the ultimate tenderness of whole tissue meat as assessed after heating were
listed by Smulders et al. (1991) and include: (1) species effects, related both to the vulnerability
of muscles to phenomena like cold shortening, but also to ageing rates (these decrease in the
order poultry > pork > lamb > beef); (2) sex, e.g. bulls generally rendering tougher meat than
cows (a muscle dependent phenomenon also partially explained by differences in fat deposition
and the age at which animals are slaughtered; see the following entry); (3) age, largely based
on a decrease in collagen solubility; (4) nutrition, energy-rich rations promoting a higher
degree of marbling; (5) preslaughter handling, possibly resulting in PSE or DFD meat; the
latter condition yielding meat with high tenderness scores, probably because calpain activity
is prolonged at high ultimate pH values; (6) rate of carcass refrigeration, possibly leading to
cold (or heat) shortening; (7) accelerated processing, e.g. by relying on electrical stimulation
possibly combined with hot boning, (8) tenderness promoting additional processing, such
as pelvic suspension or pressure/heat treatment; and finally, (9) adhering to optimal ageing
times; i.e. 5 days for pork and around 2 weeks for beef, veal, lamb and rabbit.
Not related to processing per se, but rather to domestic preparation of meat, are various
methods for cooking meat, which have a marked influence on tenderness as perceived by
the consumer. The reader is referred to Smulders et al. (1991) for further information on this
important tenderness determinant.
4.3 The water holding of meat
After the carcass and muscles are cut, a red proteinaceous fluid, called drip, oozes from the
cut surfaces. The mechanism of its formation is not completely revealed yet. Fluid loss can be
partly explained by the decreased water-binding by proteins resulting from the post mortem
pH fall. However, the amount of water chemically bound to proteins is too small to account
for the total fluid loss. The following major mechanisms of water-holding of fresh meats have
been reported and extensively discussed by Den Hertog-Meischke et al. (1997).
4.3.1 Shrinkage of myofibrils
Since most of the muscle water is present within the myofibrils, a general hypothesis for
explaining drip loss is that it originates from the ‘lateral shrinkage’ of the myofibrils post
mortem (Offer and Trinick, 1983). Lateral shrinkage is partly due to post mortem pH fall, as
with decreasing pH the charges of the filaments and thus the negative electrostatic repulsion

between the filaments are reduced, causing the space between the filaments to diminish
(Hamm, 1986).
The second factor causing (‘longitudinal’) shrinkage is the actomyosin formation during
rigor onset. The shorter the sarcomere length, the smaller the myofilament lattice spacing,
and a close and linear relationship between drip loss after one week of storage and sarcomere
shortening has been observed (Smulders et al., 1984, Honikel et al., 1986).
A third factor promoting shrinkage is the denaturation of myosin, which reduces the charge
of – and hence diminishes the electrostatic repulsion between the thick filaments as well as
reduces the myosin head length. All these events draw the thick and thin filaments close
together and thus result in higher drip loss (Offer et al., 1989).
4.3.2 Denaturation of sarcoplasmic proteins
Although it has been well-established that in meat with a low water-holding capacity
sarcoplasmic proteins are denatured, one must confront the fact that these proteins only hold
about 3% of the water and that upon denaturation these globular proteins lose their compact
folded structure and may thus entrap more rather than less water. Monin and Laborde (1985)
suggest that precipitation of (denatured) sarcoplasmic proteins may contribute to the loss of
electrostatic repulsion between filaments and thus to drip loss.
4.3.3 Changes in osmotic pressure
Maximal osmotic pressure reached in post rigor meat is highly muscle dependent and may
partly account for the variability in water-holding capacity between muscles within a carcass
(Monin and Ouali, 1991). As muscle pH drops, extracellular spaces become hyperosmotic,
causing migration from water out of the muscle cells until the sarcolemma becomes permeable
for proteins and ions.
4.3.4 Fluid accumulation within the muscle
Offer and Cousins (1992) studied the time course of structural changes in post mortem beef
sternomandibularis muscle. Immediately after the animal’s death, the extracellular space
of a muscle primarily exists of the vascular system and connective tissue (perimysium and
endomysium). After 4-6 hours post mortem, large gaps (presumably filled with fluid) appear
between the fibre bundles. After rigor onset, these gaps also arise between the fibres and the
endomysial network. The separation between fibres is only partial and a fibre may remain
in contact with one of its neighbours (see Figure 7). At this time the sarcolemma loses its
integrity and sarcoplasmic proteins can diffuse freely into the extracellular space. The fluid
in the extracellular space may be the source of drip. Both perimysial and endomysial gaps
form longitudinal channels, which are in connection with the cut surface of the meat. Drip
probably arises predominantly by the action of gravity draining the fluid in these channels
to the cut surfaces (Offer et al., 1989).

fibre
endomysium
extracellular space
perimysium
A
extracellular space
B
filled with fluid
Figure 7. Schematic overview of structural changes in beef post mortem. (A) Living muscle and immediately
after slaughter; (B) 4-6 hrs post mortem; (C) rigor (Offer and Cousins, 1992).
4.3.5 Ante mortem and processing effects on the water-holding of fresh meat – summarised
The following factors were identified by Smulders et al. (1991) and Den Hertog-Meischke et
al. (1997) as affecting the water-holding properties of fresh meat: (1) animal species and breed
differences, partly associated with their vulnerability to cold shortening and/or their (largely
genetically determined) susceptibility to stress; (2) pre-slaughter handling, possibly leading to
the PSE or DFD condition (see above); (3) stunning method (e.g. electrical stunning may cause
a somewhat accelerated glycolysis); (4) electrical stimulation, leading to faster post mortem
pH decline causing increased protein denaturation; (5) chilling rate, high chilling rates
slowing down protein denaturation but – unless counteracted by appropriate measures such as
electrical stimulation – also bearing the risk of inducing the cold shortening phenomenon; (6)
hot boning, allowing a faster and more uniform drop in temperature and thus counteracting
protein denaturation, but at the same time increasing the risk of cold shortening; (7) sample
characteristics, i.e. the degree to which muscles are cut and whether the cut is longitudinal
(less drip) or transversal (more drip); (8) method and material of support, suspended muscles
losing less than those supported from below, and muscle portions laid on a tray losing less than
those placed on absorbent paper; (9) packaging methods, the effects being largely dependent
on the forces applied on the meat and the degree to which cut ends are sealed off; (10) storage
temperature, probably because at higher storage temperatures fluids have a lower viscosity and
will migrate more easily; (11) freezing and thawing; fast freezing resulting in the formation of
many small ice crystals which cause less cell damage, and slow thawing allowing remigration
of water into the intracellular space; (12) transport conditions, transport vibrations possibly

enhancing drip loss from meats with a low intrinsic water holding capacity; and finally (13)
ageing time, extended storage periods possibly allowing re-uptake of drip as a result of pH
increase and proteolytic changes.
4.4 The flavour of meat
Flavour is determined both by taste and olfactory sensations, although astringency, mouth-
feel and juiciness also play a role. Whereas mouth receptors can assess the 4 taste sensations
(sweet, salt, sour, bitter), hundreds to thousands odour compounds can be distinguished by
epithelial receptors which are reached either by smelling (via the nose) or via posterior nares
at the back of the nose and throat while food is being chewed in the mouth (Farmer, 1994).
Moody (1983) lists the non-volatile or water-soluble compounds with taste or tactile properties
as: inorganic and sodium salts of certain acids (salty), hypoxanthine (an ATP metabolite),
peptides and some amino acids (bitter), sugars and some amino acids (sweet) and acids
(sour). In addition, there are thousands of low-molecular weight compounds that give rise
to odour sensations. Although more than 800 volatile compounds have been identified in
cooked meat aroma (Maarse, 1989), it is believed that only a relatively small number of these
compounds actually play a role in the overall aroma of cooked meat and whether one of these
represents a key odour impact compound depends on both its concentration and its odour
threshold, i.e. how sensitive the human nose is to that particular compound. Farmer (1994)
lists the major compounds and their routes of formation and she indicates two reactions that
are of particular importance in meat aroma formation, i.e. the Maillard reaction (a complex
network of reactions which yields both high-molecular-weight brown coloured products
and volatile aroma compounds) and lipid oxidation during heating which contributes to the
formation of desirable flavours. The latter author concludes that free amino acids, peptides,
sugar and also phospholipids and their fatty acids are particularly significant for flavour
forming reactions, that various vitamins and minerals can alter the rate and extent of these
reactions and consequently that changes in composition of nutrients in meat could lead to a
change in the balance of flavour forming reactions and therefore to a change in the overall
aroma and flavour.
4.4.1 Ante mortem and processing effects on the flavour of cooked fresh meat – summarised
The following factors have been identified by Smulders et al. (1991) as affecting meat flavour:
(1) animal species and breed, probably through the genetic control of lipid composition and
metabolism; (2) sex (e.g. ‘boar taint’ in pork, for which compounds such as androstenon
and skatole are thought to be responsible); (3) nutrition (e.g. ‘boiled cabbage’ off-flavours of
mutton fed with rapeseed, or ‘fishy’ off-flavours of pork fed with polyunsaturated fatty acids
which also makes this pork more prone to oxidation and hence rancidity; (4) ante mortem
stress, for instance, the DFD condition’s inherent low acidity and hence a flavour which is
less ‘accentuated’ making DFD taste rather bland; (5) bacterial contamination, as a result
of which, dependent of the bacterial ecology, ‘sulphurous’ off-flavours (Pseudomonadaceae,
Enterobacteriaceae) or ‘dairy/cheesy’ off-flavours (Lactobacillaceae, Brochothrix thermospacta)
may occur; (6) irradiation at high dosages (causing increased carbonyls or hydrocarbon
formation); (7) lipid (per)oxidation particularly catalysed by heme iron and reported in meat

that has been size-reduced (e.g. deboned chicken and turkey); (8) phospolipid and fatty acid
oxidation (‘warmed-over flavour’) in cooked meats, probably catalysed by non-heme iron.
5. Some data on game meat species
5.1 Introductory remarks
Whereas worldwide a vast number of wild animals are hunted – in situations where this is
essential for the provision of nourishment but also to satisfy man’s intrinsic desire to hunt
(Winkelmayer, 2009) – the type and range of animals hunted for food vary in different parts
of the world, depending on climate, animal diversity, local taste and locally accepted views
about what can or cannot be legitimately hunted.
Contrary to the situation for farm animal species, on which a vast body of information is
available from the literature, muscle physiological specifics of wild animals and scientific
data on their meat quality are scarce, amongst other reasons because only in rare cases (e.g.
Hoffman and Wiklund, 2006) there is an economic incentive to support domestic or export
game meat marketing by solid research. Not surprisingly, much of the information on the
sensory quality of various game species appears to be rather based on culinary tradition than
on scientific investigation.
The experimental design (particularly as regards random sampling of experimental and control
animals from a uniform population for which ample physiological and other background data
are available) is relatively easily achieved in studies on farm animal species. However, this
represents the major hurdle in studies on game species, unless these are (semi-) domesticated.
The latter is for instance the case for deer and reindeer as outlined by Wiklund and Smulders
(2011). In this context, one must also realise that once animals are domesticated, breeding
efforts may affect the animal’s physiology as evidenced, for instance, by the altered muscle fibre
structure and biochemistry of the breast muscle of domesticated turkey in comparison with
the wild counterpart animal from which it was bred (Sosnicki et al., 1991) or by the in-bred
genetic stress susceptibility prevalent in Landrace pigs (Eikelenboom and Minkema, 1974).
In the framework of this contribution it would be impossible to provide a comprehensive

overview of all the data available on game meat properties. Also, the methodologies used
by the various scientists often differ markedly, making comparison of published results
difficult. Therefore, the purpose of the following section is to provide some results of studies
on meat quality characteristics of a restricted number of game meat species as conducted in
our laboratory with the standardised methods described by Hofbauer and Smulders (2011).
Although this authorship has also been involved in studies of more exotic game animals (e.g.
on properties of guanaco- (Gonzalez et al., 2004) or beaver meat (Hofbauer et al., 2005)) we
will in the following focus on animal species with relevance to hunting activities in Central
Europe. In the absence of sufficient muscle biological background information, some of these
data are inevitably of a more descriptive nature.

5.2 Ruminant species
5.2.1 Feral deer
Table 1 and 2 include results of a study of the seasonal variation in meat quality in one year
old roe deer (Capreolus capreolus) shot (and subsequently eviscerated) in Austria’s subalpine
regions either in Spring or Autumn (Winkelmayer et al., 2004). Whereas carcass weights
differed – obviously related to the plane of nutrition available in the animals’ habitat – none
of the variables measured indicate a difference relevant for sensory quality as perceived by
the consumer.
Table 1. Carcass weight, pH and water holding capacity (WHC, method of Grau and Hamm, 1953,
procedures according to Hofmann et al., 1982) as assessed at 45-60 min (‘1’) and at 24 hrs. post mortem
in semimembranosus muscle of one year old roe deer carcasses (means ± standard deviation) (after
Winkelmayer et al., 2004)1.
Quality trait Spring (n=15) Autumn (n=12)
Carcass weight (kg) 11.07b±1.72 13.18a±1.72

pH1 6.54±0.3 6.41±0.45
WHC1 0.69±0.17 0.79±0.14
pH24 5.66±0.28 5.58±0.15
WHC24 0.72a±0.16 0.84b±0.08
1 Figures with different superscripts differ significantly (P<0.05, F- Test).
Table 2. Sensory quality traits of longissimus muscle of one-year old roe deer as assessed after 14 days of
vacuum storage at 2 °C (means ± standard deviation); none of the differences were significant (P>0.10;
F-test) (after Winkelmayer et al., 2004).
Drip loss (g/100 g) 3.11±1.13 2.35±1.05

Cooking loss (g/100 g) 18.70±3.1 19.67±2.95
Shear force (N/cm2) 23.45±4.61 22.08±4.30
Sarcomere length (µm) 1.78±0.08 1.89±0.09
L* value 37.06±3.06 36.94±2.73
a* value 14.91±1.25 15.96±2.23
b* value 10.36±1.7 11.37±1.59

5.2.2 Chamois
In Table 3 and 4 the results are presented of a similar study (Hofbauer et al., 2006) on seasonal
meat quality variation in Chamois (Rupicapra rupicapra) essentially following the same
procedures as those for roe deer.
Again, carcass weights reflected the expected effects of differences in nutritional input over
the seasons. The contrasts in colour (L*-values) are likely to have resulted from differences
in the availability of nutrients (e.g. iron), the more so because the variables related to water-
holding (drip loss) indicate that light reflection in Autumn samples is superior to that in
Spring samples. For the latter observation there is no adequate physiological explanation as
the pH/temperature profile of Autumn and Spring samples (indicative for possible differences
Table 3. Carcass weight and pH, water holding capacity (WHC; method of Grau and Hamm, 1953,
procedures according to Hofmann et al., 1982) and temperature as assessed in semimembranosus muscle
of carcasses of one-year old chamois at 45-60 min (‘1’ hr) and at 24 hrs post mortem (means ± standard
deviation) (after Hofbauer et al., 2006)1.
Carcass weight (kg) 8.39b±1.36 13.94a±2.18

pH1 6.49±0.31 6.63±0.42
WHC1 0.75±0.15 0.61±0.21
Temp1 33.70±4.44 34.15±3.09
pH24 5.61±0.23 5.84±0.54
WHC24 0.72±0.16 0.77±0.13
Temp24 5.12±2.88 7.08±3.41
1 Figures with different superscripts differ significantly (P<0.05, T- test).
Table 4. Sensory quality traits of longissimus muscle of one year old chamois, as assessed after 14 days of
refrigerated storage (mean value ± standard deviation) (after Hofbauer et al., 2006).1
Drip loss (g/100 g) 1.37a±0.72 2.29b±0.66

Cooking loss (g/100 g) 16.36±1.94 17.38±2.21
Shear force (N/cm2) 41.48±23.33 33.70±11.12
Sarcomere length (µm) 1.79±0.16 1.74±0.09
L* value 39.30a±4.43 31.00b±2.25
a* value 15.50±1.78 16.83±2.01
b* value 11.51±1.28 10.83±1.70
1 Figures with different superscripts differ significantly (P<0.05, T- test).

in the degree of myofibrillar lattice spacing or those in protein denaturation) and data on
myofibrillar density (sarcomere lengths) are similar.
5.3 Wild boar
Hofbauer and Smulders (unpublished results) investigated wild boar (Sus scrofa) meat to
determine whether its ageing rate is similar to that observed in muscles of domesticated pigs.
To this end 10 animals were killed by hunting and transported to a chill room at 0 °C. At 3
days post mortem longissimus samples were excised from the carcasses and stored overnight at
0-2 °C, after which (at 4 days post mortem) these samples were either subjected to immediate
analysis or vacuum packed and stored at 0-2 °C for a further week (i.e. 11 days post mortem).
Table 5 includes the results.
No differences of practical relevance were observed between the two storage periods. As
shear force between day 4 and 11 days post mortem was similar, it appears that – as is the
case for domesticated pigs (for which optimum ageing times between 5-7 days are reported
(Dransfield et al., 1980-1981)) – maximum tenderness is already achieved by the earliest time
wild boar meat is marketed in commercial practice. In essence, these results concur with those
reported by Zmijewski and Korzeniowski (2001). Marchiori and DeFelicio (2003) report that,
compared to commercial pork, wild boar meat has a more gradual glycolysis, lower L* and a*
colour values and (at least in female animals) lower exudative (drip) losses.
5.4 Feral pheasants
Hofbauer et al. (2010) investigated meat quality of hunted feral pheasants (Phasianus
colchicus L.). The righthandside superficial breast muscles of 14 birds were removed after 24
hrs of chilling for immediate analysis, the lefthandside counterpart muscles were left on the
refrigerated carcass until 96 hrs post mortem and then analysed. Data are presented in Table 6.
Table 5. Sensory quality traits of wild boar longissimus muscle as assessed at 4 days post mortem and
at 11 days post mortem (i.e. after one week of vacuum refrigerated storage) (Hofbauer and Smulders,
unpublished)1.
Quality trait Day 4 post mortem Day 11 post mortem
pH 5.47a±0.04 5.51b±0.04
Drip loss (g/100 g) - 6.24±3.37
Cooking loss (g/100 g) 20.90a ±2.90 22.54b±3.11
L* value 54.52 ±4.12 53.17±5.32
a* value 18.17±2.22 18.85±2.07
b* value 17.69±1.97 17.98 ±1.00
Shear force (N/cm2) 40.34±9.22 43.81±10.68
1 Figures with different superscripts differ significantly (P<0.05; T-test).

Table 6. Sensory quality traits of pheasant pectoralis superficialis muscle as assessed at 24 and 96 hrs
post mortem1 (after Hofbauer et al., 2010).
Quality trait Time post mortem
24 h p.m. (n=14) 96 h p.m. (n=14)
pH 5.62±0.05 5.64±0.08
Cooking loss (g/100 g) 11.51±4.90 12.60±1.77
Shear force (N/cm2) 28.9±13.02 31.8±11.23
L* value 54.24a±4.49 56.61b±3.45
a* value 3.81±1.86 4.04±1.43
b* value 8.02a ±1.24 9.18b±1.24
1 Figures with different superscripts differ significantly (P<0.05; T-test).
L* values are in the same range, a* and b* values higher than- and shear forces and ageing
rates similar to those recorded for other poultry species (e.g. turkey, e.g. Hillebrand et al.,
1991, Smulders and Hofbauer, 2003)
6. Conclusions
Targeted research conducted over the past decades has elucidated many of the muscle
biological mechanisms which explain the variation in meat quality, albeit primarily that
of the major farm animal species. In addition, many processing technologies allowing an
improvement of sensory quality traits have been developed and recommended standardised
laboratory methods to assess these have been made available.
Notwithstanding these achievements, one must confront the fact that publications on the
quality assessment of feral game species are scarce and the generated information is mostly
fragmented. Inevitably, available data are generally descriptive rather than interpretative.
Those factions with a vested interest in increasing consumer acceptability of game meat items
and striving to extend what is currently still a ‘niche market’ are well-advised to base their
strategies on scientific investigations, to make more research funds available allowing the
generation of relevant data and, finally, to more effectively communicate to hunters and meat
processors what needs to be done to assure the best possible sensory quality of game meat.

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Muscle biological and biochemical ramifications
of farmed game husbandry with focus on deer and
reindeer
Eva Wiklund1 and Frans J.M. Smulders2
1Svenska Samernas Riksförbund (SSR), Magasinsgatan 7, 903 27 Umeå, Sweden; eva@sapmi.se

Summary
The deer farming industry in New Zealand and the traditional Sami reindeer husbandry
culture in Fennoscandia (Sweden, Norway and Finland) lead the world in commercial venison
(deer meat) production from semi-domestic or farmed deer species, i.e. red deer (Cervus
elaphus) and reindeer (Rangifer tarandus tarandus), respectively. Production systems are
based on natural pastures but include pre-slaughter handling routines and a slaughter process
similar to that of domestic species like beef and lamb. Venison has several attributes attractive
to consumers – it is tender, has low fat content, a favourable fat composition and high levels
of minerals. All these attributes of venison are criteria demanded by today’s discerning meat
consumer. The introduction of new venison production routines such as intensive farm-based
management, industrialised slaughter and meat processing, use of commercial feed mixtures
and possibly new ingredients used to supplement or replace pasture can alter venison quality.
One topic of central importance for venison is the image as a natural, free-range origin,
clean and healthy product. Care should be taken to ensure the positive image of venison as
a ‘natural’ and healthy product is not lost when production systems are intensified to meet
the market demand.
Keywords: meat quality, venison, pH, shelf life, tenderness, colour, flavour
1. Introduction
To illustrate the worlds’ most important sources of venison (deer meat), the examples in this
chapter are taken from the young deer farming industry in New Zealand and the traditional
Sami reindeer husbandry culture in Fennoscandia (Sweden, Norway and Finland). These two
deer industries are mainly focused around venison production based on systems dependent
on pastures. The red deer (Cervus elaphus) and the reindeer (Rangifer tarandus tarandus) are
the most common deer species for venison production, in New Zealand and Fennoscandia,
respectively.
Reindeer husbandry is performed in a less intensive way than red deer farming, with the
reindeer free ranging (i.e. not enclosed in fenced areas) in forests and on the mountain tundra.

Nevertheless, at times reindeer are fed during winter to prevent starvation and to improve
body weight and condition (Staaland and Sletten, 1991). In 1986, parts of the reindeer pasture
areas in Sweden were affected by radioactive fallout from the accident in the Chernobyl
nuclear power plant, and therefore feeding is still used in these areas as a measure to reduce
radioactive caesium in the reindeer and thereby in the meat (Åhman, 1999). Also in New
Zealand, many deer farmers feed their animals a variety of supplements during winter to
provide extra nutrition when pasture is inadequate.
Consumer opinion is increasingly important to meat industries worldwide and consumers

value attributes such as flavour, tenderness and nutrient content when evaluating the quality
of meat (Smulders et al., 1991; Dransfield, 2003). Although the quality preferences of different
consumer groups may vary, producing consistent quality is critical: the quality of every
purchase should be the same. Venison is also attractive to the health-conscious consumer for its
low fat content, favourable fat composition and high mineral content (Drew and Seman, 1987).
2. Production systems for venison
2.1 Red deer
New Zealand has pioneered the development of farm-based production systems for venison.
However, deer are not native to New Zealand. The first deer were brought there from England
and Scotland for sport (hunting) in the mid-late 19th century. The environment proved ideal
and feral deer populations grew uncontrolled. By the middle of the 20th century feral deer were
regarded as a pest because of their impact on the environment and native forests. The export
of venison from feral deer started in the 1960s, turning a pest into an export earner. Industry
pioneers saw an opportunity to build on this base and in the early 1970s started capturing
live deer from the wild and farming them. A new industry was born and rapidly spread
throughout New Zealand (Deer Industry New Zealand, 2010). There are currently about 3,800
deer farms in the country, ranging in size from smaller lifestyle properties to farms carrying
many thousands of deer. On these farms there are approximately 1.7 million deer (estimated
in June 2006), or half the world’s farmed deer population. Reflecting the original imported
wild population, the majority of New Zealand’s deer herd is red deer (Cervus elaphus) (Figure
1), the balance is predominantly elk (also called ‘wapiti’) (Cervus elaphus nelson) descending
from animals imported from the USA and Canada or red/elk hybrids. There are also small
numbers of fallow deer (Dama dama).
An important component of the research and development for the New Zealand deer industry
has been to define production systems that render distinctive and high-value attributes to
venison, together with post mortem processing systems to complement these goals. The deer
industry aims to meet market demand for venison by slaughtering deer in early spring, when
animals are 9-11 months old. A major strategy is to calve as early as possible and to maximise
growth rates in the first 7 months of life before first winter. Currently, a small proportion of
animals achieve target slaughter weight pre-winter. This provides the choice to either maintain
the animals through winter (when feed is expensive) then slaughter in spring when venison

Figure 1. New Zealand has pioneered the development of farm-based production systems for venison
(Photo: Eva Wiklund).
prices are highest, or the novel opportunity to slaughter prior to winter, before the animals’
metabolism undergoes major photoperiod-induced changes (Wiklund et al., 2008).
As a specialist culinary product, venison is processed in specialised facilities licensed by the

New Zealand Ministry of Agriculture and Forestry. The amount of venison produced in New
Zealand in 2008/2009 was estimated at 21,000 tonnes (Deer Industry New Zealand, 2010). An
animal welfare Code of Practice governs the humane treatment of deer prior to processing to
ensure minimum stress and to enhance product quality. Venison is processed according to
customer requirements. The trend is towards added-value (or further processed) cuts, where
the bones and ‘silverskin’ (i.e. the connective tissue surrounding the muscles) are removed.
2.2 Reindeer
The Sami people can be regarded as the indigenous population of the Arctic region of the
Nordic countries and the Kola Peninsula. Already in 1539 the well known map of Olaus
Magnus, the Carta Marina, showed reindeer and people inhabiting this area. The oldest Sami
relics derive from the Komsa culture – a people inhabiting the region between Tromsø in
northern Norway and the Kola Peninsula in northwest Russia – as far back as 8,000 BC. The
nomadic lifestyle of the Sami people did not evolve until the late 16th century and until the 19th

century they roamed over vast expanses with their reindeer. From this perspective, reindeer
herding was very intensive and Sami families tended their reindeer herds all year round (Bäck,
1993). Scefferus (1673) reported seeing large numbers of wild reindeer in northern Sweden,
and also tame reindeer herds tended by the Sami. Tame reindeer were very useful as beasts
of burden and for milk and meat production (Figure 2).
The herders guarded their animals constantly. Winter time was the main slaughter period,
when the castrated males were in prime condition and the cold weather made it possible
to store meat over long periods. The reindeer carcass and viscera were utilised with the
utmost thoroughness, supplying the herding family with far more necessities than food
alone Beach, 1981. During the 20th century, Swedish reindeer herding has gradually become
more and more extensive. Sami families now live in Swedish communities and use all kind
of modern technology for reindeer herding, such as helicopters, airplanes, motorbikes and
snow-machines (Bäck, 1993). However, the traditional knowledge around reindeer is still very
much in focus in the Sami reindeer herders’ everyday life (Figure 3).
Traditionally, reindeer were slaughtered at the selection site, i.e. at various locations
surrounding the reindeer herding districts of the Sami people. In Sweden, the new directives
regarding reindeer meat inspection were instituted in 1993 National Food Administration,
1993) and consequently many of the former outdoor slaughter sites were closed, the numbers
of reindeer transported to slaughter increased and new mobile slaughter facilities were
developed. The rules applied for animal transport, veterinary inspection of living animals
and carcasses, stunning methods, slaughter hygiene, carcass grading and chilling conditions
for reindeer are similar to those applied for other domestic species.
Figure 2. Tame reindeer were very useful as beasts of burden and for milk and meat production for the
Sami people of northern Fennoscandia and Russia (from Scefferus, 1673).

Figure 3. The traditional knowledge around reindeer is very much in focus in the Sami reindeer herders’
everyday life (Photo: Anders Wiklund).
The Swedish reindeer population has during the last 10 years varied between 200,000-300,000
head (winter stock) and on average over this time period an annual 23% of the population
has been slaughtered. During the same period the proportion of slaughtered calves per total
number of slaughtered reindeer has increased from 43 to 67%. Due to the small production
(1,500 tonnes in 2008/2009; Sami Parliament, 2009) reindeer meat is a very exclusive gourmet
product, which is in high demand and always on the menu in the more luxurious restaurants.
Almost no reindeer meat is exported. The meat is consumed fresh but is also marketed as
cold- or hot-smoked and dried meat products.
3. Impact of production systems on venison quality
3.1 Muscle glycogen content and meat pH and their microbiological consequences
Meat pH is related to shelf life, tenderness, colour and water-holding properties, and is
therefore a good indicator of meat quality. A pH value of 5.5-5.7 is within the normal range,
while values over 5.8 result in reduced shelf life, especially for vacuum-packaged meat. Meat
with high pH, so called DFD (Dark, Firm, Dry) meat, is a persistent quality defect found in all
meat species. Meat pH values are directly correlated to the levels of muscle energy (glycogen)
at the time of slaughter (Gill and Newton, 1981; Hood and Tarrant, 1981; Tarrant, 1989). If
the glycogen stores in the muscles are low, meat pH will be elevated. Low muscle glycogen
stores might result from poor physical condition, intense physical activity or stress during
pre-slaughter handling. It has been demonstrated that deer and reindeer in good physical

condition produce meat with optimal pH values, whether they were fed a commercial feed
mixture or grazed (Wiklund et al., 1996a; Wiklund et al., 2000).
Two comprehensive surveys of meat pH for deer (n=3,600; New Zealand) and reindeer
(n=3,400; Sweden) demonstrated DFD frequencies (i.e. meat pH >6.2 measured in the M.
longissimus at the last rib) of 1.5% for red deer and 6% for reindeer (Table 1). In the New
Zealand study (Pollard et al., 1999) there was no indication of a relationship between pH and
the stress parameters studied (fighting or agitation in lairage or unsettled behaviour in the
lead-in race to stunning). It was suggested that other factors besides physical exertion at the
slaughter plant affected meat pH; for example effects of transport, yarding and handling at the
farm were not studied. On the contrary, the Swedish results (Wiklund et al., 1995; Wiklund
et al., 1996b) clearly showed that selecting reindeer for slaughter using a lasso had the most
negative effect on meat pH. In both surveys it was highlighted that there are possibilities
to improve pre-slaughter handling routines for reindeer and red deer to further reduce the
frequency of DFD carcasses.
Figure 4 and 5 (Wiklund and Smulders, 1997) illustrate the microbiological effects of
distinct ultimate pH values on shelf life and safety of reindeer meat. Whilst at 3 days post
mortem Aerobic Colony Counts (ACC) tended to be higher in high pH meat (P<0.10), all
samples reached microbial numbers around 6 log units after 2 weeks of vacuum storage.
However, at that time Enterobacteriaceae Colony Counts (ECC) of samples with high pH
were approximately one log unit higher than those with intermediate and low pH. Such
observations have been explained by Gill and Newton (1981) to be the result of low glycogen
stores favouring the predominance of proteolytic microbial flora rather than the general
positive effect high pH values have on microbial growth.
3.2 Tenderness, proteolytic enzymes and muscle fibre composition
Variation in meat pH and glycogen content can give rise to considerable variation in meat
tenderness in species such as beef and lamb (Watanabe et al., 1996; Purchas et al., 1999), and
similar results have been found for red deer venison. Within the normal pH range, values
around 5.5 have been reported to yield more tender venison than those in the 5.8-6.0 range
(Stevenson-Barry et al., 1999). This intermediate pH venison was tougher than normal pH
even after ageing and also more variable in tenderness (i.e. of less consistent quality) than
normal pH (deer) venison.
Table 1. Average ultimate pH values (pHu) and frequencies of intermediate DFD (5.8 ≤ pHu ≤ 6.2) and
DFD (pHu >6.2) in red deer, fallow deer and reindeer venison (M. longissimus).
Ultimate pH Intermediate DFD DFD (%) Reference

(overall means) (%)
Red deer (n=3,500) 5.64 9.1 1.5 Pollard et al., 1999

Fallow deer (n=100) 5.93 68.3 1.0 Pollard et al., 1999
Reindeer (n=3,400) 5.67 23.1 6.0 Wiklund et al., 1995

7
Aerobic colony count
(log10 CFU/cm2)
1
0
5,52 5,58
5,6 5,64
5,7 5,8 6,02 14 days
Low 6,05 6,09 7 days
6,23 6,45 3 days
Medium 6,53 6,87
High 7,25
pH ult
Figure 4. Aerobic Colony Count of individual reindeer longissimus samples of various ultimate pH, as
assessed after 3,7 and 14 days of refrigerated vacuum storage (Wiklund and Smulders, 1997).
7
Enterobacteriaceae colony count
6
(log10 CFU/cm )
2
0
5,52 5,58
5,6 5,64
5,7 5,8 14 days
6,02 6,05
Low 6,09 6,23 7 days
6,45 6,53 3 days
Medium 6,87 7,25
High
pH ult
Figure 5. Enterobacteriaceae Colony Count of individual reindeer longissimus samples of various

ultimate pH, assessed after 3, 7 and 14 days of refrigerated vacuum storage (Wiklund and Smulders, 1997).

Longissimus pH 7.0
high
6.5
intermediate
6.0
low
5.5
1 3 5 35
hours p.m.
high 35.4-39.0 10.3-16.2 5.9-9.6 4.9-6.7
Range in core temp. interm. 36.3-37.9 11.8-19.0 6.7-9.8 5.6-6.1
low 38.1-39.2 12.9-17.9 7.1-11.8 5.2-5.9
Figure 6. Early post mortem pH and temperature decline in lumbar longissimus muscle of 15 refrigerated
reindeer carcasses with various ultimate pH values (Smulders and Wiklund, 1998; also see Table 2).
In contrast, reindeer venison has been found to be extremely tender regardless of meat pH
(Wiklund et al., 1997). Figure 6 and Table 2 (Smulders and Wiklund, 1998) illustrate the
rate of pH and temperature decline in reindeer lumbar longissimus muscle and the effect of
ultimate pH on major physical/chemical traits related to the sensory quality of reindeer meat.
The darker appearance of DFD meat is reflected in the higher L values observed in samples
with pH >6.1. Although water-holding capacity was found to be decidedly higher than that
usually observed for ruminant meat species (i.e. drip loss values in the range of 5 to 7% as
reported by Den Hertog-Meischke et al., 1997a,b), the values for the various pH groups were
remarkably similar. However, cooking losses (indicating both the water loss from intra- and
extracellular matrix and heat-induced changes in protein structure) were significantly lower
in high pH reindeer meat as is generally the case for all meat species (Honikel, 1992). Shear
force values – although rather low in all samples regardless of pH – decreased with higher
ultimate pH and extended storage caused further tenderisation and a gradual shift in contrasts
between the various pH groups.

Table 2. The effects of ultimate pH of reindeer longissimus muscle on physical-chemical variables related
to eating quality (n=5 in each subgroup; least square means and standard error); (After Smulders and
Wiklund, 1998).
Trait Low pH Medium pH High pH
Minolta L*
3 days 30.1a ±0.9 26.0b1±1.2 26.5b±0.9
7 days 30.4a±0.9 5.7b12±1.2 26.8b±0.9
14 days 30.3a ±0.9 25.1b2±1.2 26.7b±0.9
Minolta a*
3 days 11.4±1.0 13.0±1.3 11.2±1.0
7 days 11.5±1.0 13.1±1.3 10.8±1.0
14 days 11.4±1.0 13.1±1.3 11.3±1.0
Minolta b*
3 days 5.8±0.8 5.1±1.0 3.8±0.8
7 days 6.2a±0.8 5.3ab±1.0 3.7b±0.8
14 days 1.0±0.2 5.2ab±1.0 3.9b±0.8
Drip loss (%)
3 days 1.1±0.1 1.1±0.2 1.1±0.1
7 days 1.2±0.1 0.9±0.2 1.1±0.1
14 days 1.1±0.2 0.9±0.2 1.1±0.2
Cooking loss (%)
3 days 18.8a±2.3 14.0ab±3.0 10.4b±2.3
7 days 17.8a±2.4 14.6ab±3.0 10.6b±2.3
14 days 18.6a±2.4 14.1ab±3.2 11.8b±2.4
Shear force (kg/cm2)
3 days 5.3a1±0.6 4.8a1±0.6 2.4b±0.6
7 days 4.0a2±0.7 3.8ab12±0.6 2.1b±0.6
14 days 4.0 a2±0.6 3.1ab2±0.6 2.0b±0.6
Within rows, means with superscripts not containing a common letter differ significantly (P<0.05); Within columns
and traits, means with superscripts not containing a common number differ significantly.
The observation that shear force values of venison are relatively low (indicating high
tenderness) has been confirmed in more recent studies on both red deer by Farouk et al.
(2009) and on reindeer by Rincker et al. (2006), who report that venison is generally much
more tender than beef, which is explained both by its small muscle fibre size (Taylor et al.,
2002) and its higher activity of proteolytic enzymes as compared with beef (Barnier et al.,
1999; Farouk et al., 2007a) (Table 3).
In all species raised for meat production, genetic selection and improvement in nutrition and
breeding conditions has led to a tremendous increase in the efficiency of animal production
and improvement in carcass composition by decreasing fatness and increasing muscle yield
(Lefaucheur, 2010). Comparisons between wild and domesticated animals within different

Table 3. Tenderness (shear force) measured in beef, red deer, fallow deer and reindeer M. longissimus in
non-electrically stimulated carcasses from animals aged 1.5-2 years.
Shear force (kgF) Shear force (kgF) Reference

1-3 days post mortem 1 week post mortem
Beef (n=8) 11.7 9.8 Barnier et al., 1999

Red deer (n=7) 11.4 8.2 Wiklund et al., 2001a
Fallow deer (n=8) 5.4 Sims et al., 2004
4.5 Volpelli et al., 2003
Reindeer (n=8) 2.9 2.6 Wiklund et al., 1997
species suggest that selection for increased growth rate and lean meat content has shifted
muscle metabolism/fibre type composition towards a more white/glycolytic and less red/
oxidative type (Ashmore, 1974; Ruusunen and Puolanne, 2004). White/glycolytic muscle fibres
are bigger than red/oxidative fibres, and several studies have found a negative relationship
between increased muscle fibre size and meat tenderness for a number of animal species
(Lefaucheur, 2010).
A pilot study was carried out to compare quality attributes in meat from fast growing
young red deer stags slaughtered prior to winter (which had reached their slaughter weight
at 7 months of age) with that of slower growing young red deer stags slaughtered more
traditionally, at 12 months of age (Wiklund et al., 2008). Overall, this pilot study highlighted
relatively minor differences in quality attributes of venison from fast and slow growing
red deer stags, indicating that as the deer farming industry heads towards more efficient
venison production systems there are unlikely to be any major negative impacts on product
quality (Wiklund et al., 2008). However, further studies of the impact of growth rate on
venison processing, packaging and storage techniques should be conducted before making
recommendations to the venison industry. We are not aware of any further studies on how
the selection for increased growth rate and lean meat content in deer would affect deer muscle
fibre composition – and possibly venison tenderness – but it is reasonable to believe that
intensified genetic selection for meat production would eventually affect deer in a similar
way to the other animal species studied.
3.3 Meat colour and colour display life
Colour is an important attribute affecting the decision by the consumer at the point of purchase
whether to buy meat on retail display or not (Bekhit and Faustman, 2005). Consumers judge
the acceptability of meat colour by how bright red the meat looks on display, and a strong
correlation exists between redness (a*), hue angle and consumer colour acceptability (Farouk
et al., 2007b). Hue angle is also a good indicator of colour stability of meat on display. The
browning of meat, which determines the colour display life, is due to the reaction where red
oxymyoglobin is oxidised to brownish metmyoglobin. The speed of this oxidation process

is dependent on several factors like antioxidant content, oxygen consumption and reducing
enzyme activity in the meat (Faustman and Cassens, 1990).
Venison contains a higher concentration of myoglobin (Young and West, 2001) and pro-
oxidants such as iron and copper (Drew and Seman, 1987; Stevenson-Barry et al., 1999)
than beef. This fact probably explains the overall poorer colour of venison relative to beef.
In addition, the higher enzymatic activities of venison compared to beef may have resulted
in venison losing its metmyoglobin reducing activity, resulting in faster and irreversible
oxidation than usually observed in beef and consequently leading to a decreased colour
display life (Farouk et al., 2007a). Wiklund et al. (2006) demonstrated a significantly better
colour display life of red deer and fallow deer (Wiklund et al., 2005) venison from grazing
animals compared to deer fed a pelleted feed or whole barley, and suggested that this difference
was related to the higher content of natural antioxidants (vitamin E) in the pasture compared
to the grain-based feed (Figure 7 and 8). In a recent study it was suggested that the colour
display life of red deer venison was related to a seasonal variation in pasture quality (Wiklund
et al., 2010). Meat from animals slaughtered in the spring had a significantly better colour
stability compared with meat from animals slaughtered in the summer, autumn and winter
120
110
100
90
80
Display life (hours)
70
60
50
40
30
20
10
0
0 1 3 6 12
Weeks post-slaughter
Figure 7. Mean display life (hours of Minolta a* value ≥12) in M. longissimus from the red deer (Cervus
elaphus) from two treatments (• pasture grazing and ■ pellet fed, n=8 in each group) included in the
study, measured at 1 day, 1, 3, 6 and 12 weeks of refrigerated storage (-1.5 °C), with error bars indicating
standard error of difference (S.E.D) (from Wiklund et al., 2006).

300
250
200
150
100
50
** **
0
0 1 7 14 21
Days post-slaughter
Figure 8. Mean display life (hours of Minolta a* value 12) in M. longissimus from the fallow deer (Dama
dama) from two treatments (• pasture grazing and ■ barley fed, n=12 in each group) included in the study,
measured at 1 day, 1, 2, and 3 weeks of refrigerated storage (+2 °C), with error bars indicating standard
error of difference (S.E.D) (from Wiklund et al., 2005).
which was explained by a higher antioxidant content of the superior quality pasture growing
in the spring.
3.4 Chemical composition and flavour
Natural or managed pastures (grasses, herbs and bushes) contain high levels of poly-
unsaturated fatty acids (PUFA) and are also rich in different antioxidants. Grain-based feeds
are higher in saturated fatty acids (SFA), and antioxidants like vitamin E are often added to
commercial feed mixtures. When animals are grazing pasture or if they are fed grains, the
fatty acid composition in their muscles/meat will change towards the composition of their feed
(Wood and Smulders, 1999; Wood et al., 2008). Fatty acid profiles in red deer and reindeer
venison related to feed-type have been thoroughly investigated and linked to venison flavour.
Similar results have been found for both species; deer/reindeer grazing pasture produced
PUFA-rich venison with more ‘grassy’, ‘gamey’ and ‘wild’ flavours, while the grain fed animals
gave meat with significantly less PUFA and a ‘mild’ and ‘beef-like’ flavour (Wiklund et al.,
2003a,b) (Table 4). These flavour differences were demonstrated using both trained sensory

Table 4. Mean values for fatty acid composition (% of total fatty acids) in M. longissimus from pasture
and pellet-fed reindeer (Rangifer tarandus tarandus) and red deer (Cervus elaphus).
Fatty acid Reindeer1 Red deer2
Pasture Pellets Degree Pasture Pellets Degree

(n=6) (n=9) of sign.1 (n=7) (n=7) of sign.3
Polar lipids
SFA4 25.4 26.3 n.s. 25.9 24.4 n.s.
MUFA 17.3 16.0 * 13.8 12.4 n.s.
PUFA (n-6) 31.9 39.4 *** 29.3 41.9 ***
PUFA (n-3) 14.2 7.5 *** 14.2 4.5 ***
(n-6)/(n-3) 2.2 5.3 *** 2.1 9.6 ***
Neutral lipids
SFA 53.0 54.6 n.s. 54.7 50.6 **
MUFA 37.6 39.2 * 36.4 39.8 *
PUFA (n-6) 2.6 2.3 n.s. 4.3 6.6 **
PUFA (n-3) 1.4 0.3 *** 2.5 0.6 ***
(n-6)/(n-3) 1.9 8.1 *** 1.7 12.3 ***
1 Wiklund et al., 2001b.
2 Wiklund et al., 2003b.
3 n.s. = P>0.05; * = P≤0.05; ** = P≤0.01; *** = P≤0.001.
4 Saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids.
panels and consumer tests. New formulations for grain-based reindeer feed mixtures, where
ingredients like linseed cake (Sampels et al., 2006) and fish meal (Finstad et al., 2007) were
included, verified that the higher PUFA content in the feed was reflected in the meat, so that
the fat composition of reindeer venison from animals fed the new formulations was very
similar to that of reindeer venison from grazing animals.
From a human health perspective, a reduction in SFA and an increase in PUFA intake have
been recommended to combat cardiovascular disease and cancers. Within PUFA, the omega-3
fatty acids are preferred over the omega-6 because they confer positive nutritional and
physiological advantages (Williams, 2000). Today’s Western diets have an omega-3/omega-6
ratio between 15 and 20, whereas it is assumed that it was close to 1 during human evolution
(Simopoulus, 2002). From a nutritional point of view it is suggested that the omega-3/omega-6
ratio should be below 4, and it is therefore important to include sources rich in omega-3
PUFA in the daily diet. The venison industries are in a unique position to take advantage of
the fact that grass-fed venison has naturally high levels of omega-3 PUFA and antioxidants
(e.g. vitamins C and E, carotenoids and flavonoids).

4. Ethics and image of venison
Production systems like reindeer husbandry and deer farming, where the animals graze
during most of the year, are usually considered more animal-friendly and ethical compared
with the standard commercial production of beef, pork or poultry and venison is a product
that meets most of the criteria demanded by today’s discerning meat consumer (Hoffman and
Wiklund, 2006). Most of the basic principles (effects of gender, age, region, etc.) and practices
(nutrition, pre-slaughter handling, transport, lairage, stunning, electrical stimulation, etc.)
that influence meat quality and composition that are applicable to more traditional red meat
species are also applicable to deer (Hoffman and Wiklund, 2006).
In more intensive production systems the use of strategic feeding and new feed ingredients,
herd health vaccination programmes, artificial insemination and even embryo transfer can
be a part of normal practise. Some studies have reported on the use of growth hormones
to improve deer productivity (Mulley et al., 1996), although hormones to increase meat
production have been totally rejected by the deer industries world-wide. It is logical to wonder
whether including too many of these ‘standard commercial production practises’ in the deer
farming systems will in the long run damage the consumers’ perception of venison being
different from other types of meat. Care should be taken to ensure the positive image of
venison as a ‘natural’ and healthy product is not lost when production systems are intensified
to meet the market demand. In addition, the deer industries use the natural, healthy and
ethical image as their central message in any promotion and marketing of venison.
5. Conclusions
The introduction of new venison production systems such as intensive farm-based

management, industrialised slaughter and meat processing, use of commercial feed mixtures
and possibly new ingredients used to supplement or replace pasture can alter venison quality.
One topic of central importance for venison is the image as a natural, free-range origin, clean
and healthy product. Concerns about how this image could change when new feeding regimes
are introduced have to be balanced against the need of using these feeds as supplements or
replacements when pastures cannot provide enough nutrition for maintenance and growth.
The deer studies from New Zealand and the reindeer studies from Sweden suggest that there
may be several pre-slaughter conditions that could be improved for deer and reindeer, leading
to a more consistent venison quality. It is also of interest for the venison industries world-wide
to recognise the quality differences related to different feeding regimes. For example, venison

with more or less ‘wild’ flavour could be directed to specific markets based on information
about production system/feeding regime and consumer preference.
Acknowledgements
The authors would like to acknowledge the following research teams which all have contributed
to the research results presented in this paper: the Meat Science group and the Reindeer
Husbandry Unit at the Swedish University of Agricultural Sciences, Uppsala, Sweden; the
Department of Food of Animal Origin, Faculty of Veterinary Medicine at Utrecht University,
the Netherlands, the Department of Production Animal Medicine and Veterinary Public
Health, University of Veterinary Medicine at Vienna, Austria, the Reindeer Research Program
at the University of Alaska Fairbanks, USA; the Deer Systems group and the Meat Science team
at AgResearch Ltd., Invermay and Hamilton, New Zealand and the Deer Unit at the University
of Western Sydney, Richmond, Australia. We are also grateful for positive contacts with
reindeer herders, deer farmers and meat processors and the industry organisations particularly
Svenska Samernas Riksförbund (SSR) in Sweden and Deer Industry New Zealand (DINZ) and
resulting discussions and input which have significantly contributed to the research.
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Other contributions

A summary of methods to assess major physical-
chemical and sensory quality traits of fresh (whole
tissue) meat
1210 Vienna, Austria; peter.hofbauer@vetmeduni.ac.at
Summary
This contribution summarises the standard methodologies used to assess meat quality
characteristics as applied at the University of Veterinary Medicine Vienna, Austria. It also
compares the utility and validity of the various approaches to measure sensory quality of fresh
meat in scientific experiments and indicates where major references for further consideration
can be found.
Keywords: meat quality, fresh meat, sensory traits, methodologies
1. Introduction
Product quality as defined by ISO is ‘the totality of features and characteristics of a product
that bear on its ability to satisfy stated or implied needs’ (ISO, 1986). Relevant to the sensory
quality of meat are only those features and characteristics that determine the acceptability
by the consumer (Smulders et al., 1991). By using physical or chemical methods or relying
on the senses of analytical (‘expert’) panels, the major sensory quality traits can be measured
in numerical terms. Apart from its relevance for predicting how successful (acceptable) a
meat item may potentially be on the consumer market, this serves the important purpose of
identifying the biological mechanisms that underlie the variability in quality characteristics
of fresh meat of various animal species as influenced by fresh meat processing (see Hofbauer
and Smulders, 2011). In the following, the major methods commonly used in meat quality
research laboratories to assess the sensory quality of meat are summarised.
2. pH and temperature decline
The pH of post mortem muscle is a central characteristic explaining a number of muscle

biological events. Both the speed of early post mortem pH decline (in concert with
the temperature at which this takes place as this, for instance, determines the muscle’s
vulnerability to protein denaturation) and the ultimate pH value (dependent on residual post
mortem glycogen stores) are important indicators explaining possible effects on water-holding
capacity, colour and tenderness. Hence these variables are recorded in meat research, as well

as for purposes of (on line) industrial quality management. Generally, either the traditional
glass- or more robust steel-covered pH probes (e.g. the K21 electrodes allowing up to 600
measurements per hour at temperatures ranging from 0-80 °C and hence more suitable for
on-line use) and digital steel thermometers are applied for this purpose.
3. Waterholding capacity (drip loss; cooking loss)
It is crucial to distinguish the concepts of ‘water-holding’ and ‘water-binding’ capacity.

Whereas the former is relevant to assess the muscle’s intrinsic ability to hold on to its own
water, the water bound in the course of meat products manufacture (i.e. generally implying
the addition of salts in products to be cooked) reflect the potential of the muscle matrix to
bind added water (brine).
Various methods exist to quantify water-holding such as the ‘press-method’ (i.e. pressing a
standard meat sample between filter paper, leaving a ring of water the diameter of which is
related to water-holding as described by Grau and Hamm (1953); see Figure 1), centrifugal
methods (at 5.000 to 40.000 g), capillary suction by gypsum discs as in Hofmann’s capillary
volumeter (Hofmann, 1975, particularly suitable to distinguish PSE from normal meat),
measuring weight increases of analytical filter paper placed on the meat’s surface (Kauffman
et al., 1986) or even through costly NMR analysis (Den Hertog-Meischke, 1997).
The utility of the various methods has been extensively discussed by Trout (1988). For
instance, whereas the press method appears to be very practicable, its accuracy is not, as
Figure 1. Measurement of the water holding capacity of fresh meat by the press-method (Grau and
Hamm, 1953).

A summary of methods to assess major physical-chemical and sensory quality traits
such is heavily dependent on the homogeneity of the rather small size muscle sample and
related to difficulties in ensuring standardisation of the pressure applied. Consequently,
recent years have seen most researchers refraining from this simple piece of equipment and
rather relying on procedures recommended by an OECD working group (Barton-Gade et
al., 1993). These are based on weight loss measurements over a standard time (usually 24 hrs)
with muscle samples being suspended in an inflated polyvinyl bag at 5 °C, carefully avoiding
the sample making contact with the bag (Figure 2). The same sample can be used for further
drip loss measurements (e.g. after 2, 7 days, etc.), but in every case the initial sample weight
is used as reference point.
Heating of fresh meat results in water (‘cooking’) loss from the intra- and extracellular
muscle matrix (i.e. through cell membrane damage, shrinkage of muscle fibres, aggregation of
sarcoplasmic proteins and particularly by shrinking of connective tissue; Offer, 1984). Honikel
(1998) describes the standard procedure for measuring cooking loss in whole tissue meat.
It relies on placing a weighed and standardised meat slice of max. 50 mm thickness in thin-
walled plastic bags in boiling water until the sample’s internal temperature has risen to 75 °C
while making sure the bag opening extends above water level (Figure 3) and subsequently
cooling down the sample in an ice slurry and holding it in chill conditions (1 to 5 °C) until
equilibrated, whereupon the sample is blotted dry and reweighed. Occasionally water bath
temperature is held at maximum of 80 °C (in such cases sometimes the term ‘heating loss’ is
used) to facilitate keeping sample core temperatures at maximum 75 °C (e.g. Eikelenboom
and Smulders, 1986).
Figure 2. Assessment of water holding capacity by measuring weight loss over time in suspended fresh
meat samples.

Figure 3. Assessment of cooking loss by measuring weight differences before and after heating in a water
bath.
Water-holding capacity being heavily dependent on the integrity of muscle proteins (as
for instance markedly influenced by the PSE condition in meat), sometimes additional
chemical tests are performed. These include determining the degree of protein solubility
with classical or more modern methods or by measuring bound phosphorylase (Hart, 1962;
Den Hertog-Meischke et al., 1997). These additional tests allow for distinguishing protein
denaturation effects from those caused by other chemical or physical events and thus enable
a more mechanistic interpretation of the results. In this context it should also be noted that
information on the density of the muscle matrix (measurable by assessing sarcomere length;
see below) is important to determine the physical potential of meat to contain (‘structurally
bound’) water.
4. Fresh meat colour
Colour as perceived by the consumer is the physical modification of light by colorants as

observed by the human eye and interpreted by the brain. For a person to see the colour of an
object there needs to be light illuminating, and being reflected from, that object. Absolute
colour scores cannot be provided by visual assessment since humans have a poor colour
‘memory’. Guidelines for human evaluation of meat colour have been published by AMSA
(1991).
To monitor changes in meat colour over time, instruments are commonly used to measure
reflection and absorption. The principles of these ‘objective’ colour measurements have been

described by Klettner and Stiebing (1980). The frequently used, so-called CIELAB system is
based on the L*, a*, b* colour space suggested by the Commission Internationale de l’Eclairage
(CIE, 1976). In this system L* denotes the lightness (black/white) coordinate, a* the red/
green coordinate and b* the blue/yellow coordinate. Mathematical transformations of these
coordinates, notably Chroma (or saturation, i.e. the square root of (a*)2+(b*)2, indicating the
degree to which colour differs from grey) and Hue (or colour tint, i.e. the tangent of the ratio
b*:a*, describing the names of the colours red, via yellow and green, to purple) describe colour
changes in more imaginable terms (Figure 4).
The following factors determine muscle colour variation by affecting light absorption, i.e.
(1) pigment content (notably Myoglobin); (2) oxygenation and oxidation of Mb (resulting in
formation of Oxymyoglobin and Metmyoglobin, respectively) and by affecting light reflection
i.e. (3) water on the meat’s surface, which is dependent on its water-holding properties (see
Hofbauer and Smulders, 2011)
The recommended procedure for the instrumental measurement of fresh meat colour in
numerical terms has been described by Honikel (1998). It includes casting light from a defined
source (D65 with illumination/viewing system as 45/0 or 0/45 or diffuse /8 (d/8)) on the
meat surface with a preferred observer angle of 10° and relying on the L*a*b* colour scale.
Apart from ensuring that the measuring instrument (e.g. Hunter or Minolta equipment) is
adequately calibrated using black (L* = 0) and white (L* = 100) colour standards, the meat
must be allowed to ‘bloom’ for 1-2 hrs prior to measurement so as to ensure its surface is fully
oxygenated (Figure 5).
Standardised colour scales with an appearance mimicking a meat surface and with a 1-6
scale have also been made available for qualitative visual colour classification to be applied
on-line, such as the Japanese pork colour scale developed by Nakai et al. (1975), allowing for
a rapid distinction between PSE, normal and DFD pork (Figure 6).
white
lightness
–a
green +b
ation yello
w
satur
–b +a
blue red
colour tint
black
Figure 4. The colour space: L*, a* and b* values (Smulders, 1986).

Figure 5. Colorimetry of fresh meat cut surface after 1-2 hours of exposure to air.
Figure 6. Visual scale for measuring fresh pork colour (Nakai et al., 1975).
PSE: Pale-Soft-Exudative; DFD: Dark-Firm-Dry.
5. Tenderness traits
To study the significance of ante and post mortem factors affecting tenderness methods are
required that allow some sort of quantification of the effects. In marketing and trading of
carcasses and fresh meat determining the ‘coarseness of grain’ (i.e. the fibre size as assessed
by running the ball of the thumb over a transverse muscle section and feeling its smoothness)
is still often used by ‘experts’ as a tenderness indicator. Although such a procedure has
been suggested to be related with tenderness (Hammond, 1952) it is extremely subject to
personal interpretation. Moreover, to distinguish the distinct roles of the various tenderness
determinants fine-tuned methodologies are required.
Many mechanical devices have been developed to simulate the shearing, penetrating, biting
mincing and compression actions of the human teeth. Pearson (1963) lists their inherent

errors and limitations and suggests that the correlation coefficient between shear values and
sensory tenderness of the widely used Warner-Bratzler shearing device (found in various
studies to vary between 0.60 and 0.85 (average 0.75)) is quite satisfactory, considering the
variability within sensory panels alone.
The Warner-Bratzler device consists of a 1.2 mm thick stainless steel blade with a hole in which
the sample is placed. The blade is led through a slid between two shear bars and the amount of
shearing force is recorded with the aid of a draw bench (e.g. the Instron apparatus; see Figure
7). More closely related to the biting action of the teeth is the Volodkevich device. That consists
of rounded wedges or bars between which the meat sample is squeezed and sheared until it
finally breaks. The so-called ‘initial yield’ (the first inflexion in the force-deformation curve,
which, however, is not always clearly observable) is generally associated with the myofibrillar
contribution to tenderness, whereas the ‘peak force’ reflects the combination of myofibrillar
and connective tissue strength (Figure 8). The MIRINZ tenderometer used in most studies
from New Zealand appears to primarily measure the former.
Honikel (1998) discusses the recommended procedures in using both these pieces of equipment.
They include detailed descriptions of muscle selection and preparation (e.g. preferably lumbar
longissimus muscle), muscle excision (perpendicular to the muscle’s longitudinal axis), muscle
size (at least a 50 mm slice), heating procedure (in essence as described under waterholding;
see above), number of core samples to be tested (at least 10), fibre orientation and size of core
samples (10×10 mm cross section strips of at least 30 mm length with fibre direction parallel
to longitudinal axis) and the draw bench’s shearing speed and direction (at 50-100 mm/min
at right angles to the fibre axis).
Figure 7. Draw bench fitted with a Warner-Bratzler shear device.

PFIY
Force
Peak force
Initial yield
Distance
Figure 8. Force deformation curve of the Warner-Bratzler shear force measurement (Honikel, 1998).
Usually, concurrent with mechanical shearing, additional tests are conducted which assess
the influence on tenderness of muscle contraction. As muscle contraction may considerably
increase myofibrillar density (and consequently the substance to be sheared in a core sample),
sarcomere length is measured, either microscopically (a rather time-consuming procedure)
or via laser diffraction (Figure 9). The recommended procedures for the latter method have
been described by Koolmees et al. (1986).
Such and further tests (for instance those indicative for the degree of meat ageing, or even more
specifically for the activity of the enzymes and their inhibitors responsible for proteolysis)
are helpful when one is particularly interested in why meat is tender or tough rather than
how tender or tough it is.
‘Subjective’ measurements of tenderness, i.e. by a panel of individuals relying on their

physical senses, are often renounced because the training and testing of the panellists is
time-consuming and costly. Yet, instrumental analysis of tenderness cannot be considered
a complete substitution for subjective evaluation. Should one consider that taste panel
studies are necessary, it is essential to realise that for its reliable quantification by panellists
only trained experts should be relied on, Provided these are available their number can
be limited. In ‘consumer acceptance studies’ (i.e. assessing the marketing possibilities of a
certain product) trained panellists are useless as the purpose of the exercise is to invite the
more qualitative opinions of inexperienced consumers representing a true cross section of
the target group. For consumer acceptance studies a panel size of at least 50 individuals has
been suggested. A detailed description of recommended procedures for sensory panel studies
on meat and meat products has been made available by Cross et al. (1978).

Figure 9. Measuring sarcomere length (SL) by laser diffraction. A Helium-Neon laser beam (wave length
632.8 nm) is sent through a fibre specimen (fixed by glutaraldehyde);
-3 2 2
SL (in μm) = 632.8×10 ×√(D +T )
T
6. A remark on problems associated with sample size: animal species

differences
Whereas most of the procedures described above are indeed suitable for use in major
production animal species such as beef, lamb and pork, some animal species simply do not yield
enough muscle substance to perform all the tests according to the above recommendations
so choices on the variables to be tested and/or numbers of animals necessary for a justified
experimental design must be made. This is particularly relevant in the sensory analysis of
small (including game) birds. Also, since – in contrast to the economically important muscles
from mammalian species – avian musculature has a very distinct distribution of fibre types
(compare the almost entirely white breast muscle with the largely red leg musculature) results
should be interpreted with care.
References
AMSA (American Meat Science Association), 1991. Guidelines for Meat Color Evaluation. AMSA/National
Livestock and Meat Board, Chicago, IL, USA.
Barton-Gade, P.A., Demeyer, D., Honikel, K.O., Joseph, R.L., Puolanne, E., Severini, M., Smulders, F.J.M. and
Tornberg, E., 1993. Final version of Reference methods for water-holding capacity in meat and meat Products;
Procedures recommended by an OECD Working Group. Proc. 39th ICoMST, Calgary, Canada, File S4, PO2.WP.
CIE (Commission Internationale de l’Eclairage), 1976. Colorimetry, 2nd Edition. CIE, Vienna, Austria.

Cross, H.R., Bernholdt, H.F., Dikeman, M.E., Greene, B.E., Moody, W.G., Staggs, R. and West, R.L., 1978. Guidelines
for the Cookery and Sensory Evaluation of Meat. AMSA/National Livestock and Meat Board, Chicago, IL,
USA, 25 pp.
Den Hertog- Meischke, M.J.A., Klont, R.E., Smulders, F.J.M. and Van Logtestijn, J.G., 1997. Variation in rate of
glycolysis does not result in variation in water-holding capacity of moderately chilled, non-stimulated veal
longissimus muscle. Meat Sci. 47, 323-329.
Eikelenboom, G. and Smulders, F.J.M., 1986. The effect of electrical stimulation on veal quality. Meat Sci. 16,
103-112.
Grau, R. and Hamm, R., 1953. Eine einfache Methode zur Bestimmung der Wasserbindung im Muskel.
Naturwissenschaften 40, 29.
Hammond, J., 1952. Objective tests for quality in meat. Ann. de la Nutrit. et Alim., Paris VI, C 119-131.
Hart, P.C., 1962. Fysisch-chemische kenmerken van gedegenereerd vlees bij varkens. Tijdschr. Diergeneesk. 87, 156.
Hofbauer, P. and Smulders, F.J.M., 2011. The muscle biological background of meat quality including that of game
species. In: Paulsen, P., Bauer, A., Vodnansky, M., Winkelmayer, R. and Smulders, F.J.M. (eds.). Game meat
hygiene in focus: Microbiology, epidemiology, risk analysis and quality assurance. Wageningen Academic
Publishers, Wageningen, the Netherlands, 273-295.
Hofmann, K., 1975. Ein neues Gerät zur Bestimmung der Wasserbindung des Fleisches: das ‚Kapillar-Volumeter’.
Fleischwirtschaft 1, 25-30.
Honikel, K.O., 1998. Reference Methods for the assessment of physical characteristics of meat. Meat Sci. 49, 447-457.
ISO, 1986. ‘ISO-8402’. Quality Vocabulary. International Organization for Standardization, Geneva, Switzerland.
Kauffman, R.G., Eikelenboom, R.G., Van der Wal, P., Merkus, G. and Zaar, M. (1986). The use of filter paper to
estimate drip loss in porcine musculature. Meat Sci. 18, 191.
Klettner, P.G. and Stiebing, A., 1980. Beitrag zur Bestimmung der Farbe bei Fleisch und Fleischerzeugnissen. 1.
Einführung in die Grundlagen der Farbmessung. Fleischwirtsch. 60, 1970-1976.
Koolmees, P.A., Korteknie, A.F. and Smulders, F.J.M., 1986. Accuracy and utility of sarcomere length assessment
by laser diffraction. Food Microstr. 5, 71-76.
Nakai, H., Saito, F., Ikeda, T., Ando, S. and Komatsu, A., 1975. Standard models for pork colour. Bull. Nat. Inst.
Animal Industry 29, 69-74.
Offer, G., 1984. Progress in the biochemistry, physiology and structure of meat. Proc. 30th Eur. Meet. Meat Res.
Work., Bristol, UK, pp. 87-94.
Pearson, A.M., 1963. Objective and subjective measurements for meat tenderness. Proc. Meat Tenderness
Symposium, Campbell Soup Co., Camden, NJ, USA, p. 135.
Smulders, F.J.M., 1986. Sensory meat quality and its assessment. Veterinary Quarterly 8, 158-167.
Smulders, F.J.M., Van Laack, R.L.J.M. and Eikelenboom, G., 1991. Muscle and Meat Quality; Biological Basis,
Processing, Preparation. In: Smulders, F.J.M. (ed.). The European Meat Industry in the 1990’s. Publ.
ECCEAMST/Audet, the Netherlands, pp. 121-159.
Trout, G.R., 1988. Techniques for measuring water-binding capacity in muscle foods – A review of methodology.
Meat Sci. 18, 281-293.

Evaluation of some parameters of post mortem
changes of pheasant (Phasianus colchicus)
Jozef Nagy1, Peter Paulsen2, Peter Popelka1, Peter Lazar1, Jaroslav Soroka1,
Valent Ledecký1, Slavomír Marcinčák1, Beáta Korénekova1 and Pavel Bystrický1
1The University of Veterinary Medicine and Pharmacy in Košice, Komenského 73, 041 81
Košice, Slovak Republic; nagy@uvm.sk

Summary
Post mortem changes were evaluated in muscle tissues of 99 male, uneviscerated pheasants
(Phasianus colchicus) stored for up to 14 days at 0 and 4 °C. The control group (Group I)
included regularly slaughtered and bled pheasants (n=33). From approximately 350 pheasants
hunted on a single occasion, specimens were taken and, on the basis of X-ray examination
performed from latero-lateral and dorso-ventral body projection of hunted pheasants, two
groups were constituted: Group II included pheasants (n=33) with one or multiple shots in the
muscle tissues but not in the body cavity, whereas Group III included pheasants (n=33) with
shots in both muscle tissues and the body cavity. Each group was divided into two subgroups,
in which uneviscerated pheasants were stored at 0 °C and 4 °C, respectively, for a maximum
of 14 days. Randomly selected pheasants from each subgroup (n=4), were examined on days
0, 3, 7, and 14. The following post mortem changes in breast and thigh muscles were evaluated:
pH value (in watery extract), lactic acid and ammonia concentration. Generally, low pH values
were associated with higher concentrations of lactic acid, most pronounced in breast muscle.
Significant differences (P<0.05) in pH – both at 0 and 4 °C storage temperature – were found
for breast muscles between Groups I and II, albeit average differences were merely in the range
of merely 0.1 to 0.2 units. Thigh muscles of Group I had significantly lower pH (0.2 to 0.4
units) than Groups II and III, for all study days. A statistically significant difference (P<0.05)
was found between the lactic acid concentrations in breast muscles (4 °C) between groups II
and III. The highest average concentrations of ammonia in the breast muscle (day 14) were
determined in the Group III (4 °C), and the lowest in the group of slaughtered pheasants
(4 °C). The highest average concentrations of ammonia in the thigh muscles were in Group
III (0 °C), and the lowest in the group of slaughtered pheasants (0 °C). This difference was
statistically significant (P<0.05). In summary, muscles from pheasants being slaughtered
instead of hunted were characterised by lower pH, higher lactic acid and lower ammonia
concentration at the end of the storage period.
Keywords: pheasant, post mortem changes, lactic acid, pH, ammonia

Jozef Nagy et al.
1. Introduction
The conversion of muscle to meat has been extensively described by Hofbauer and Smulders
(2011). It starts by the conditioning period, i.e. the time period between slaughtering of the
animal and the onset of rigor mortis, during which muscle glycogen reserves are broken down
anaerobically, resulting in the intramuscular accumulation of lactic acid and the concurrent
decline in pH values. During the subsequent ageing period muscle proteolysis occurs, which
may after extended ageing involve the formation of alkalic substances such as ammonia with
concurrent elevation of pH values (Lawrie and Ledward, 2009; Hofbauer and Smulders, 2011).
In this context, these events to some extent reflect the progress of proteolytic changes.
By measuring intramuscular pH changes and simultaneously assessing the lactic acid

and ammonia generation one may monitor glycolytic and proteolytic changes. Measuring
intramuscular pH values by probes is not uncomplicated, as the interface of the probe’s acid-
sensitive membrane and the muscle surface liquids may vary, dependent for instance on the
location of measurement. For example, subsequent pH measurements on the same site where
the initial incision was made occasionally generate slightly different pH values than those
conducted at fresh incision sites. To limit such effects, intramuscular pH is alternatively
measured in watery suspensions of muscle samples taken from the carcasses under scrutiny
(e.g. Smulders et al., 1990).
The objective of this study was to compare post mortem changes (pH, lactic acid, ammonia) of
exsanguinated or hunted pheasants over a 14 day storage period at two different temperatures,
i.e. 0 and 4 °C.
2.1 Origin of the pheasants
The post mortem changes were evaluated in 99 pheasants (Phasianus colchicus) from the
hunting grounds of the University of Veterinary Medicine and Pharmacy in Košice, 66
of which were randomly collected from approximately 350 pheasants hunted on a single
occasion. Animals were hunted or slaughtered according to local animal welfare regulations.
2.2 Constitution of the experimental groups
The study design has been described earlier (Paulsen et al., 2008). In brief, male pheasants were
divided into three basic groups. The control group (Group I) included regularly slaughtered
and exsanguinated pheasants (n=33) (Figure 1). Hunted pheasants were subjected to X-ray
examination in latero-lateral and dorso-ventral body projection and assigned to one of the
following groups. Group II included pheasants (n=33) with one or multiple shots in the
muscle tissues, but none in the body cavity (Figure 2), group III included pheasants (n=33)
with shots in both muscle tissues and the body cavity (Figure 3). Each group was divided into
two subgroups, i.e. pheasants stored uneviscerated at 0 or 4 °C, for a maximum of 14 days.

Figure 1. Pheasant after evisceration – Group I (slaughtered pheasants).
Figure 2. X-ray and pheasant after evisceration – Group II (shots only in muscle tissue).
Figure 3. X-ray and pheasant after evisceration – Group III (shots in both muscle tissue and body cavity).

Jozef Nagy et al.
On days 0, 3, 7, and 14, pheasants from each subgroup (4 animals) were chosen randomly,
and breast and thigh muscles were removed for laboratory examinations described below.
2.3 Evaluated parameters
Post mortem changes in the muscles of pheasants were evaluated with respect to pH value
(in watery extract), amount of lactic acid, and ammonia content. The pH values of breast
and thigh muscles (in watery extract) were measured by digital pH meter and electrode
(both WTW, Germany). The amount of lactic acid was determined by methods of analytical
capillary isotachophoresis (Isotachophoretic analyser ZKI – 001, Labeco Spišská Nová Ves,
The Slovak Republic). Ammonia content was determined by Conway’s method (Conway,
1947). The obtained results were tested by one way ANOVA. Comparison of slaughtered and
hunted groups was performed using t-test.
3.1 pH
The average pH values measured in the breast muscles were lower than the values in the thigh
muscles. This confirms previous reports from Richter et al. (1992), Paulsen et al. (2008) and
Hofbauer et al. (2010). The latter authors hypothesised that this might be due to thigh muscles
of flightless birds being more active than wing muscles, and consequently that ante mortem
activity will leave less carbohydrate for post mortem lactic acid formation.
At day 0, pH values were similar in all three groups. During storage, the highest average pH
values (breast muscle) were found in the Group III (4 °C). Marked differences in ultimate pH
values at day 14 were not observed. The highest average pH values (day 14) determined in the
thigh muscles were found in Group II (4 °C), the lowest in Group I (0 °C).
Significant differences (P<0.05) between pH values in breasts muscles of Group I animals

(slaughtered pheasants) and those of Group II (without shots in body cavity) were found. Also,
pH values of the thigh muscles stored at both 0 and 4 °C differed significantly for Group I as
compared to Group II, and Group I compared to Group III (Table 1).
3.2 Lactic acid
In breast muscles lactic acid concentrations and pH values were highly correlated at both
storage temperatures: at day 3 pH values decreased as lactic acid concentration increased,
at day 7, pH values increased and lactic acid concentration decreased, at day 14, both values
changed negligibly. Average concentrations of lactic acid were higher in breast than in thigh
muscles. At day 14, the highest average concentration in the breast muscles was measured in
Group I (4 °C), the lowest levels in Group III (0 °C).

Table 1. Comparison of average pH values in breast and thigh muscles of pheasants stored uneviscerated
at 0 °C and 4 °C, respectively.
Group I Group II Group III
0 °C 4 °C 0 °C 4 °C 0 °C 4 °C
Breast muscle
Day 0 5.79 5.79 5.76 5.76 5.84 5.84
Day 3 5.71 5.61 5.68 5.55 5.62 5.82
Day 7 5.98 5.72 5.90 5.65 6.00 5.71
Day 14 5.82 5.84 5.67 5.72 5.66 5.83
Thigh
Day 0 6.12 6.12 6.61 6.61 6.68 6.68
Day 3 6.15 6.15 6.61 6.57 6.57 6.49
Day 7 6.31 6.13 6.58 6.50 6.59 6.37
Day 14 6.04 6.21 6.64 6.45 6.36 6.53
In thigh muscles, the highest average lactic acid concentration was determined in Group I
(4 °C), the lowest one in Group II (0 °C). At day 14, lactic acid concentrations were statistically
different in the breast muscles (4 °C) of Group II (without shots in the body cavity) vs. Group
III (shots in the body cavity) (Table 2).
Table 2. Comparison of average lactic acid values (g/kg) in breast and thigh muscles of pheasants stored
uneviscerated at 0 °C and 4 °C.
0 °C 4 °C 0 °C 4 °C 0 °C 4 °C
Breast muscle
Day 0 2.13 2.13 2.13 2.13 2.47 2.47
Day 3 5.20 2.60 5.15 2.93 4.60 3.60
Day 7 3.00 3.48 3.16 3.12 4.10 3.35
Day 14 2.45 3.65 2.02 2.93 1.70 3.27
Thigh
Day 0 1.57 1.57 1.60 1.60 1.57 1.57
Day 3 4.20 2.50 3.00 2.27 2.95 2.60
Day 7 1.98 2.12 2.13 1.86 3.10 2.45
Day 14 1.70 1.80 1.20 1.77 1.10 1.33

Jozef Nagy et al.
3.3 Ammonia
The highest average concentrations of ammonia in the breast muscle (day 14) were determined
in Group III (4 °C), and the lowest in Group I (4 °C). At day 14, the highest average
concentrations of ammonia in the thigh muscles were in Group III (0 °C), and the lowest in
Group I (0 °C). Statistically significant differences were observed between Groups I and III
(thigh muscles) stored at 0 °C (Table 3).
Table 3. Comparison of ammonia content (mg/kg) in breast muscles of pheasants stored uneviscerated
at 0 °C and 4 °C.
0 °C 4 °C 0 °C 4 °C 0 °C 4 °C
Breast muscle
Day 0 3.00 3.00 3.13 3.13 3.05 3.05
Day 3 2.30 2.48 2.53 2.69 2.83 2.83
Day 7 3.78 3.99 3.88 4.11 5.14 4.03
Day 14 3.58 3.31 3.78 4.13 3.52 4.15
Thigh
Day 0 2.58 2.58 3.28 3.28 3.02 3.02
Day 3 2.09 2.62 2.55 2.54 2.79 2.47
Day 7 4.37 4.67 4.49 3.9 5.32 4.21
Day 14 3.57 3.64 4.64 4.21 6.88 4.28
4. Conclusions
Our results indicate that post mortem changes in pheasant meat are influenced by method of
killing and storage conditions. In summary, muscles from pheasants being slaughtered instead
of hunted were characterised by lower pH, higher lactic and lower ammonia concentration
at the end of the storage period.
Acknowledgements
This work was funded, in part, by the Hunters Association of Lower Austria (Vienna) and the
Österreichische Nationalbank-Jubiläumsfonds (Vienna, Austria), grant No. 11307. Thanks are
due to Prof. Dr. R. Winkelmayer, Veterinary Administration Lower Austria (Bruck/Leitha)
and Dr. M. Vodnansky, Central European Institute for Ecology of Wild Game Vienna-Brno-
Nitra (Brno, Czech Republic), for their valuable comments on the study design.

References
Conway, E.J., 1947. Microdiffusion analysis and volumetric error. Crosby Lockwood & Sons, London, UK.
Hofbauer, P. and Smulders, F.J.M., 2011. A summary of methods to assess major physical-chemical and sensory
quality traits of fresh (whole tissue) meat. In: Paulsen, P., Bauer, A., Vodnansky, M., Winkelmayer, R. and
Smulders, F.J.M. (eds.). Game meat hygiene in focus: Microbiology, epidemiology, risk analysis and quality
assurance. Wageningen Academic Publishers, Wageningen, the Netherlands, 315-324
Hofbauer, P., Smulders, F.J.M., Vodnansky, M., Paulsen, P. and El-Ghareeb, W.R., 2010. A note on meat quality
traits of pheasants (Phasianus colchicus). Eur. J. Wildl. Res. 56, 809-813.
Lawrie, R.A. and Ledward, D.A., 2009. Lawrie’s Meat Science. 7th Edition, Woodhead Publishing, Great Abington,
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Richter, G., Ochrimenko, C. and Gruhn, K., 1992. Zusammensetzung und Qualitätsparameter von Perlhühnern,
Fasanen, Tauben, Cairina und Kaninchen. Nahrung 36, 543-550.
Smulders, F.J.M., Marsh, B.B., Swartz, D.R., Russel, R.L. and Hoenecke, M.E., 1990. Beef tenderness and sarcomere
length. Meat Sci. 28, 349-363.

Biographies of key contributors
Niels Bandick, Wissenschaftlicher Oberrat Dr.med.vet., started his career in 1989 as a
veterinary surgeon. Between 1991 and 1993 he focussed his research activities on psycho-
neuro-immunology in rats, at the Hannover Medical School, and from 1993 until 1995
on game meat hygiene at the Hannover Veterinary School, Department of Meat and Food
Hygiene. Subsequently, until 2000 he conducted studies on risk-based visual meat inspection
at Bonn University, Department of Agriculture. From 2000 to 2008 Dr. Bandick was senior
lecturer and scientist at Berlin´s Free University, Faculty of Veterinary Medicine, Insitute of
Meat Hygiene and Technology, whereafter he accepted a position at the Federal Institute for
Risk Assessment (BfR), where he is currently head of the working group Food Technology
within the Department for Biological Safety (Unit Food Hygiene and Safety Concepts).
Johan L. Bekker, currently a principal lecturer in the Department of Environmental Health

with the Tshwane University of Technology in Pretoria, has been actively involved in the
training of undergraduate and post graduate students who specialise in the field of Food
Hygiene and Safety and has a special interest in meat safety throughout the meat supply chain.
Although he has been supervisor for 6 successful Masters Degree candidates he is currently
involved in a Doctoral study.
Alessandro Bianchi graduated in 1994 in Veterinary Medicine from the University of Milan,
Italy, and holds a specialist degree in Applied ethology and welfare of livestock and pet
animals and a specialist degree in Technology and pathology of avian species, rabbit and
game, both from the University of Milan, Italy. He has been working since 2005 as Head
Veterinarian in the Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia
Romagna (IZSLER), in the provincial laboratory of Sondrio. His professional activity focuses
on wild animal ethology and animal welfare, natural resource management, wild animal
clinical medicine, and zoonoses.
Katharina Brugger graduated from the University of Vienna, Austria, with a Mag. (MSc)
and Dr. rer. nat. (PhD) in Meteorology in 2004 and 2008, respectively. Since 2009 she is
Assistant Professor at the Institute for Veterinary Public Health at the University of Veterinary
Medicine Vienna. Her current research focus is on simulation of mosquito-borne diseases
in the context of climate change. In 2008 she established a mosquito monitoring for the
investigation of bluetongue disease vectors (Culicoides spp.).
Sava Buncic obtained his veterinary (DVM) as well as MScVM (i.e. MPhil) and DScVM
(i.e. PhD) degrees at the University of Belgrade (Serbia). He held positions of Assistant- and
Associate Professor in Meat Hygiene at the Faculty of Veterinary Medicine, University of
Belgrade (Serbia) until 1993. Subsequently, he was Senior Researcher in Food Microbiology
and Safety at the Meat Research Institute (now AgResearch) in Hamilton, New Zealand (1993-
1998). Between 1998 and 2006, he was Senior Lecturer in Veterinary Public Health at the School
of Veterinary Sciences, University of Bristol (UK). From 2006 and presently, he is Full Professor
in Meat Hygiene and Safety at Department for Veterinary Medicine, University of Novi Sad
(Serbia). Prof. Buncic is Diplomate of the European College of Veterinary Public Health
(ECVPH) and served as the Chair of its Education Committee. He was/is member of Editorial

DOI 10.3920/978-90-8686-723-3, © Wageningen Academic Publishers 2011
Biographies of keynote contributors
Boards of several scientific journals (e.g. J. Food Prot., Foodborne Path. & Dis., Animal).
Furthermore, Prof. Buncic has been a member of the EU-EFSA’s BIOHAZARD Scientific Panel
from its start in 2003 and holds now his third mandate for the period 2009-2012.
Armin Deutz graduated in veterinary medicine in 1986 in Vienna, Austria. In 1986 he

obtained his doctoral degree with a thesis on mastitis detection, and 2001 a PhD, discussing
a thesis on zoonosis in wildlife. From 1987 till 1990 he was affiliated with the Medical Clinic
for Ruminants and Swine at the University of Veterinary Medicine in Vienna. From 1991
to 1992 he was a private veterinary practitioner. Since 1992 he is a Veterinary officer in the
Styrian Provincial Government. He became Diplomate of the European College of Veterinary
Public Health (ECVPH) in 2004. His special interests include zoonoses, infectious diseases,
influences of climate change on the distribution of animal diseases, wildlife and game meat.
Massimo Fabbi graduated in 1983 in Veterinary Medicine from the University of Milan,
Italy, and holds a specialist degree in Veterinary Public Health from the University of Milan,
Italy. He has been working since 1985 as Head Veterinarian in the Istituto Zooprofilattico
Sperimentale della Lombardia e dell’Emilia Romagna (IZSLER), in the provincial laboratory
of Pavia. He is currently Director of the Diagnostic Departments of Pavia, Milano and
Varese of IZSLER. Since 1999, he has been the Head of the National Reference Laboratory for
Tularemia. Since 1997, he has been Lecturer in zoonoses, infectious diseases and foodborne
diseases in the Faculty of Veterinary Medicine and the School of Specialization in Veterinary
Public Health of the University of Milan. His professional activity focuses on the laboratory
diagnosis of several zoonotic diseases, mainly tularemia, cat-scratch disease, Lyme borreliosis,
chlamydiosis, leptospirosis, toxoplasmosis, brucellosis, Q fever, legionellosis, and food
microbiology.
John Fletcher graduated BVMS from Glasgow University in 1970. Subsequently, he studied the
reproductive physiology and behaviour of red deer on the the Isle of Rum, Scotland receiving
a PhD from Cambridge University in 1974. He established Europe’s first commercial venison
unit in Auchtermuchty, Scotland in 1973. John Fletcher was the first vice chairman of the
British Deer Farmers’ Association and was subsequently chairman. He was instrumental in
founding and is past president of the Federation of European Deer Farmers’ Associations and
the British Veterinary Deer Society. He has contributed to many scientific journals including
Nature and chapters to several academic books. He has written widely for the general public
in journals such as the New Scientist, Country Life, The Times, and published a full length
book, Fletcher’s Game in 2001. A further book on the history of British deer parks is in press.
Matteo Frasnelli graduated in 1990 in Veterinary Medicine from the University of Bologna,
Italy, and holds a specialist degree in Technology and pathology of avian species, rabbit and
game from the University of Milan, Italy. He has been working since 1997 as Head Veterinarian
in the Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna (IZSLER),
in the provincial laboratory of Ravenna. His professional activity has focused for many years
on the laboratory diagnosis of animal infectious diseases, including wildlife.

Alessandra Gaffuri graduated in 1986 in Veterinary Medicine from the University of

Milan, Italy, on a thesis on the infectious keratoconjunctivitis of chamois, and holds a
specialist degree in Veterinary Public Health from the University of Milan, Italy. She has
been working since 1988 at the Istituto Zooprofilattico Sperimentale della Lombardia e
dell’Emilia Romagna (IZSLER), where she currently is Head of the Diagnostic Laboratory
in the provincial laboratory of Bergamo. Her professional activity has focused for many
years on the epidemiology, diagnosis and control of infectious diseases of wildlife and small
ruminants. Since 1997, she has been responsible for an ongoing monitoring programme of
wildlife diseases in the territory of the province of Bergamo with particular focus on diseases
of chamois, roe deer, deer and wild boar. Her research activity focuses on the epidemiology
and diagnosis of tuberculosis in wildlife. In 2006-2007 she collaborated with EFSA as a
Member of a Working Group set up for assessing the suitability of existing tests for the
diagnosis of tuberculosis in deer. She has participated as a speaker in several national meetings
and international congresses on the management and diseases of wildlife.
Johann Gasteiner studied Veterinary Medicine in Vienna and received his doctoral degree
in 1997, with a thesis on a serological survey of paratuberculosis in Austrian cattle. In 1995
he joined the scientific staff of the Institute of Ruminants and Swine of the University of
Veterinary Medicine in Vienna. In 1999 he moved to the Federal Research and Education
Centre for Agriculture Raumberg-Gumpenstein and since 2005 he is head of the Institute for
Farm Animal Welfare and Animal Health. He became Diplomate of the European College of
Bovine Health Management (ECBHM) in 2006. Main topics of his work are surgery, animal
care and welfare, but also feeding and metabolic diseases in ruminants, and their impact on
animals’ health and fertility. Special topics are the development, the improvement and the
implementation of wireless sensor systems for intraruminal measuring of parameters like
temperature and pH. He is author or co-author of several publications concerning endoscopy,
enzootic calcinosis, paratuberculosis and rumen acidosis. He is also involved in training of
veterinarians, farmers and hunters.
Thomas Guggenberger was born in 1971 in Carinthia. He studied Geo-Informatics at

Johannes Kepler University Linz and at Paris Lodron University Salzburg (MSc.). Since 1992
he is member of the scientific staff at the Federal Research and Education Centre Raumberg-
Gumpenstein and since 2007 he is head of the Department of Economy and Resource
Management. His main topics are analyses of landscape changes, modelling of diseases
spread and calculations of agricultural ecobalances.
Andreas Hensel graduated in Veterinary Medicine at the Tierärztliche Hochschule Hannover

(TiHo, Germany) in 1985. He continued his studies (‘Aufbaustudium’ in microbiology) and
received a doctorate degree in 1998. From 1998-1990 he was research scientist at the Institute
of Microbiology and Infectious Animal Diseases at this Faculty. In 1990 he joined the Institute
of Bacteriology and Animal Hygiene at the University of Veterinary Medicine in Vienna and
received a PhD. Degree at Utrecht University in 1994. From 1994 until 1997 he was a senior
scientist at the Institute of Microbiology and Genetics, Biocenter, University of Vienna. After
having received his ‘Habilitation’ degree at the TiHo in Hannover in 1997, Dr. Hensel was
appointed as Full Professor for ‘Animal Hygiene and Federal Animal Pest Control’ and head

of the Institute of Animal Hygiene and Veterinary Public Health at the University of Leipzig
(1998-2008). Since 2003 he is President of the Federal Institute for Risk Assessment in Berlin
and Germany´s representative in the scientific council (Advisory Forum) of the European
Food Safety Authority (EFSA).
Peter Hofbauer studied Veterinary Medicine in Vienna, and received his doctoral degree in
1994. His research at the Institute of Meat Hygiene of the University of Veterinary Medicine
in Vienna focuses on the sensory meat quality and instrumental measurement of fresh meat
quality traits in farm as well as game animal species.
Louw C. Hoffman is Professor in Meat Science and currently Head of the Department of
Animal Science, Stellenbosch University. He has been actively involved in the training of post
graduate students and his research focus is on factors influencing the meat quality of exotic
game and ostrich meat. He has published over 125 internationally peer reviewed papers and
has received numerous awards for his research contributions.
Piet J. Jooste is Professor in the Department of Biotechnology and Food Technology, Tshwane
University of Technology in Pretoria. Previously he held the positions of Professor and Head
of the Department of Food Science at Free State University and Deputy Director of the
Animal Products and Nutrition Institute of the Agricultural Research Council in Pretoria.
His research relates to food safety, quality and hygiene and he has published more that 75
peer reviewed scientific articles. He has been supervisor or co-supervisor for 25 successful
Masters and 6 Doctoral degree candidates.
Liesel A. Laubscher is a postgraduate student who holds a joint degree in nature conservation
and animal sciences. Her research focuses on applied wild animal behaviour as pertaining
to the commercial harvesting of wild game species.
Simone Magnino graduated in 1987 in Veterinary Medicine from the University of Milan,
Italy, on a thesis on tularemia, and holds a specialist degree in Medical Statistics from the
University of Pavia, Italy. He has been working since 1990 as Head Veterinarian in the
provincial laboratory of Pavia of the Istituto Zooprofilattico Sperimentale della Lombardia e
dell’Emilia Romagna (IZSLER). His main fields of interest are the conventional and molecular
diagnostics of infectious and parasitic diseases of livestock and companion animals, and
bacterial and parasitic zoonoses. Since 1999, Dr Magnino has been Head of the National
Reference Laboratory for Animal Chlamydioses, and from 2003 until 2009, he was Member
of the Scientific Panel on Biological Hazards of the European Food Safety Authority (EFSA).
In November 2009 he moved to the Department of Food Safety and Zoonoses, WHO
Headquarters in Geneva, Switzerland, where he will be dealing with issues related to zoonoses
and the new strategy of WHO for zoonotic public health risks arising at human-animal-
ecosystem interface.
Giuseppe Merialdi graduated in 1996 in Veterinary Medicine from the University of Parma,
Italy, and holds a specialist degree in Animal Health, Rearing and Animal Husbandry
Production from the University of Parma, Italy. He has been working since 2000 as Head

Romagna (IZSLER), in the provincial laboratory of Reggio Emila. Since 2008, he has been
Director of the provincial laboratory of Bologna of IZSLER. His professional activity has
focused for many years on the laboratory diagnosis of animal infectious diseases (mainly
bacteriological diseases), food microbiology and food safety.
Milorad Mirilovic obtained his veterinary (DVM) as well as MSc (i.e. MPhil) and DScVM
(i.e. PhD.) degrees at the University of Belgrade (Serbia). In 1992 he became research assistant
at the Department of Biostatics and Economics, Faculty of Veterinary Medicine, University
of Belgrade (Serbia). Currently, he holds a position of Assistant Professor in Biostatistics. Dr.
Mirilovic published 83 scientific papers.
Maria Pacciarini is Doctor in Biological Sciences, and holds a specialist degree in Microbiology
and Virology. She has been employed at the Istituto Zooprofilattico Sperimentale della
Lombardia e dell’Emilia Romagna (IZSLER) since 1988, and is currently Head of the Molecular
Diagnostic Laboratory. She has expertise in the development and application of molecular
biology techniques for detection, identification and characterisation of pathogenic organisms
of veterinary interest; in particular she has been involved in the molecular diagnosis and
genetic differentiation of M. bovis since 1997. She has been responsible for several research
projects financed by the Italian Ministry of Health since 1997 on the development of molecular
methods for epidemiological studies of bovine tuberculosis (TB), the identification of TB
complex and atypical mycobacteria, the improvement of immunological test for diagnosis
of TB in vivo. Since January 2000, she has been Director of the National Reference Center for
Bovine Tuberculosis by Mycobacterium bovis. She has been member of the subgroup of the
TB Task Force of the European Commission since 2001.
Peter Paulsen studied Veterinary Medicine in Vienna, and received his doctoral degree
in 1994. Since that year, he has been affiliated with the Institute of Meat Hygiene of the
University of Veterinary Medicine in Vienna. He became assistant professor at this Institute
and Diplomate of the European College of Veterinary Public Health in 2003. Main topics
of work are the relation of chemical and microbiological conditions in raw materials (of
animal origin), and their impact on final food/feed quality; also, the use of ‘rapid’ or novel
microbiological methods in food or feed industry, esp. for assessment of raw material quality.
The hygiene and microbiology of meat from hunted game is a particular subject of study,
with special respect to the effect of standardised processing techniques and GHP. He is also
involved in training of hunters and trained persons. His activities for game meat research
and training of hunters were acknowledged in 2009 by the CIC literary prize.
Milorad Radakovic, BVSc, CertVPH(MH), MRCVS, graduated in Zagreb, Croatia in 1989.

After a brief period of work in Croatia he moved to the UK, where he has spent most of
his veterinary career in the Veterinary Public Health (VPH) sector. His work experience
varies from practical on-line meat hygiene inspection to performing the duties of an Official
Veterinary Surgeon (OVS) and Principal Official Veterinary Surgeon (POVS) for the Meat
Hygiene Service (MHS). He currently works as a Veterinary Adviser at the Food Standards
Agency in London, providing veterinary public health advice to policy makers and liaising
with other government departments, academic institutions and other countries. Milorad
has given numerous lectures and presentations on veterinary public health in the UK and

abroad and has appeared as an expert veterinary witness in several high profile court cases.
He is a visiting lecturer at Cambridge Veterinary School and an active Council member of
the Veterinary Public Health Association.
Franz Rubel was born in Austria, in 1962. He received his Mag. (MSc) and Dr. rer. nat. (PhD)
degrees in Meteorology from the University of Vienna, Austria, in 1989 and 1994, respectively.
In 2002 he got a position as Associate Professor (A.Univ.-Prof.) at the University of Veterinary
Medicine Vienna. His recent research focuses on epidemic modelling in veterinary sciences,
especially the simulation of the dynamics of mosquito-born diseases associated with climate
change and the airborne spread of foot-and-mouth disease virus. In 2007 he was appointed
president of the Austrian Society for Meteorology. In 2008 he established the section
epidemiology of the Austrian Society for Veterinary Medicine.
Diana L. van Schalkwyk, is completing her PhD on the game meat production in Namibia.
She has experience in the development of food safety programmes for commercial red meat
abattoirs with a special interest in game processing plants.
Frans J.M. Smulders studied veterinary medicine at Utrecht University (DVM 1978).
Following a short period in companion animal practice he joined the research staff of the
Department of the Science of Foods of Animal Origin of Utrecht University. After having been
awarded a PhD degree in Meat Science from Utrecht University in 1984, he joined the Muscle
Biology Laboratory of the University of Wisconsin at Madison as a Postdoctorate Fellow in
1986, studying the biophysics of post-mortem muscle contraction. Upon returning to The
Netherlands in 1987 he became Senior Researcher and Leader of Utrecht University’s Muscle
Biology and Meat Quality research programme. He was initiator and General Manager (1990-
1996) of the European Consortium for Continuing Education in Advanced Meat Science and
Technology (ECCEAMST). In 1996 Dr. Smulders was appointed Professor of Meat Hygiene,
Meat Technology and Food Science at the University of Veterinary Medicine at Vienna. He
served as a member of the European Commission’s ‘Scientific Committee on Veterinary
Measures relating to Public Health’ (SCVMPH; 2000-2003), was a founding father of the
European College of Veterinary Public Health (ECVPH, of which he is a registered Diplomate)
and served as its President (2000-2003) and Senior Vice-President (2004-2005). From 2005
to date Prof. Smulders has been active as a risk assessor in working groups instituted by the
Animal Health and Welfare (AHAW) and Biohazards (BIOHAZ) Panels of the European
Food Safety Agency (EFSA) at Parma, Italy. In May 2008 he was awarded an honorary
doctorate degree (Dr. h.c.) from the University of Helsinki. As of July 2009 Frans Smulders
serves as a member of EFSA’s Scientific Committee.
Peter-Vitus Stangl attended a higher technical school for agriculture and then studied
Veterinary Medicine. He received his doctoral degree in 1979. His professional career started
at the Universities of Agriculture and Veterinary Medicine, and he gathered additional
experience as a veterinary practitioner and official veterinarian in a district slaughterhouse.
Since 1985, he is affiliated with the Ministry of Health, with a focus on meat hygiene and
since 1988 holds the position of the head of the division for food safety for meat production,
primary production and animal by-products. In this position, he was actively involved in the

implementation of EU directive 92/45/EEC in Austria. He designed a three-step inspection

system which was later adopted in EU food hygiene legislation. Other duties include: chairman
of the Austrian standing committee of hygiene and member of the chapter ‘meat and meat
products’ of the Austrian Food Codex.
Eva Wiklund graduated from the Swedish University of Agricultural Sciences in 1990 and
then obtained a PhD in Food Science/Meat Science from the same university in 1997. She
has specialised in venison (deer meat) research for her entire research carreer and her work
has been very much focused around contacts with the industry, meaning the meat industry
in general and reindeer/deer industry in particular. This research profile has provided the
opportunity to build contact net-works with the world’s most important venison industries
and deer researchers in Fennoscandia, New Zealand, Australia and Alaska. Over the last 10
years she has worked for extended periods in all these countries. In 2009 she moved from
her position as a Senior Meat Scientist at AgResearch Limited, New Zealand to the Swedish
Reindeer Association (Svenska Samernas Riksförbund, SSR). She also holds a position as an
Affiliate Associate Professor at University of Alaska Fairbanks, USA.
Rudolf Winkelmayer is a veterinarian who graduated from the University of Veterinary

Medicine from which he received the Doctorate degree in 1981. From 1981 to date he combines
a private small animal practice (recognised as Specialist since 1988) with a function as Official
Veterinarian in Lower Austria. He is a licensed hunter and has initiated and continues
to coordinate transnational scientific projects on wildlife and game (e.g. since 2000 the
‘INTERREG’ project on the prevalence of Fascioloides magna in Austria, Hungary, the
Czech Republic and Slowakia), to which end he cooperates with various universities in these
countries in the fields of game meat hygiene, game diseases and animal welfare. In 2000, he
received the Josef Bayer-award from the University of Veterinary Medicine at Vienna for his
contributions to the promotion of game meat hygiene. In 2005, he was certified as Diplomate
of the ECVPH. In 2006, Dr. Winkelmayer was given the title ‘Professor’ by the Federal
Austrian Government, and in 2009 he became Chief Veterinary Counseler (‘wirklicher
Hofrat’). Over the past decades Rudolf Winkelmayer has been active in the training of
(thousands of) hunters, for which purpose he has authored or co-authored a large number
of books, some of which have been translated in various languages. For their book Wildbret-
Hygiene (Winkelmayer, Paulsen, Lebersorger, Zedka) Winkelmayer et al. received, in 2009,
the Literary Prize of the ‘International Council for Game and Wildlife Conservation’ (CIC).
Maria Grazia Zanoni graduated in 1994 in Veterinary Medicine from the University of
Milan, Italy, and holds a specialist degree in Hygiene and Technology of milk and dairy
products from the University of Milan, Italy. She has been working since 1999 as Head
Romagna (IZSLER), in the central diagnostic laboratory of Brescia. Her professional activity
focuses on the diagnosis of animal bacterial and parasitic diseases, with special reference to
the diagnosis of infections by Mycobacterium bovis.

Index
A B
abattoir (game meat establishment) 53 background toughness 282
abdominal shots 23, 82 – See also: gut backyard farming 149, 154
shots baits 256
ACC – See: Aerobic Colony Counts BASC – See: British Association for
actin 275 Shooting & Conservation
adenosine diphosphate (ADP) 276 bear 225
adenosine monophosphate (AMP) 278 best practice 22, 113
adenosine triphosphate (ATP) 276, 278 biological hazards, dog bites 212
ADP – See: adenosine diphosphate black grouse 157
Aerobic Colony Counts (ACC) 116, 302 – bleeding 82, 128 – See
See also: Total Aerobic Counts (TAC) also: exsanguination
African experience 41, 67 – throat slitting 48
Agamodistomum suis 119 – See blesbok 68
also: Alaria alata boma harvesting 75
AGHE – See: approved game handling bovine tuberculosis (BT) 45, 200
establishment – compatible lesions 201, 202
aging 278 – National Eradication Programme,
agricultural schools, infrastructure 265 Portugal 203
Alaria alata British Association for Shooting &
– life cycle 119, 120 Conservation (BASC) 216
– paratenic hosts 119 Brucella 223, 227
ammonia 326 – abortus 234
– change during storage 330 – canis 234
– pheasant, breast muscles 330 – ceti 234
AMP – See: adenosine monophosphate – in wildlife 234
animal disease control – melitensis 234
– GPS-GIS use 200 – microti 234
animal disease prevention – neotomae 234
– actions 170 – ovis 234
animal-to-fork pathway 218 – pinnipedialis 234
animal welfare 101 – species 234
– Code of Practice, New Zealand 299 – suis 234
ante mortem stress 87, 109 – transmission to humans 234
antibacterial activity 25 brucellosis 45, 200
antioxidants 308 BSE 25
approved game handling establishment BT – See: bovine tuberculosis
(AGHE) 113, 115, 116 bushmeat 60
arbovirus – See: arthropod-borne diseases bushveld 75
arthropod-borne diseases 159, 173, 174
– Germany 160 C
ATP – See: adenosine triphosphate cadmium 87
Austria 19 calpains 283
Austrian food codex 20 calpastatin 283

Index
Campylobacter 212 – fresh meat, stability 280

– jejuni 218 – meat 289, 290, 291, 306
captive bolt stunning 115 – metmyoglobin 278
capture boma 75 – myoglobin 278
capture myopathy 76, 77 – objective measurement 318
carcass damage 73 – oxymyoglobin 278
carcasses colour display life
– bruised 49 – fallow deer 307
– dressing 86 – natural antioxidants 307
– partially dressed 50 – red deer 307
– wild game, surface microflora 27 conditioning 278
cathepsin 284 conservation 71
cattle skin 25 consumer 41, 59
CCP’s – See: Critical Control Points – acceptance studies 322
Ce.R.M.A.S – See: National Reference – ethical and sustainable food
Centre for Wildlife Diseases production 245
Certificate of Origin 54 – number exposed when hazardous
Certificates of Acceptability 54 carcass processed 246
chamois 157 control 71
– Lombardy, Italy 225 conventional hunting
Chernobyl 298 – day time 77
chilling 50 – little disturbance 77
classical swine fever 200 – selective harvesting 77
clean-as-you-go 84 cooling temperatures 21
cleaning body cavities 21 Cori cycle (gluconeogenesis) 277
climate change 157, 224 CP – See: creatin phosphate
– Francisella tularensis 161, 162 creatin phosphate (CP) 277, 278
– loss of habitats for wild game 165 Critical Control Points (CCP’s) 129
– parasites 160 cross-contamination 116
– risk factors 158 customer malpractices 60
– Tyndall Centre for Climate Change
Research dataset 185 D
– wild animals species affected 159 dark, firm and dry meat (DFD meat) 71,
climate models 159, 176 137, 279, 284, 286, 301
– global 173 – cooking losses 304
climate-warming scenarios 173 – deer 302
Clostridium botulinum 218 – reindeer 302
CNS spread 25 – shear force 304
cold shortening 282 – tenderisation 304
collagen – water-holding capacity 304
– distribution 282 day harvesting 74
– solubility 282, 284 – age classes 74
colour – ante mortem stress 74
– CIELAB 319 – bruising 74
– factors affecting 280 – dermal abrasion 74
– fresh meat 278 – distinguishing sexes 74

Index
– social groupings 74 Escherichia coli 24, 25, 26, 27, 95, 107
deep muscle tissues 26 – enterohaemorrhagic (EHEC) 25, 95,
deer 107
– farming 310 – enterohaemorrhagic (EHEC),
– farming, New Zealand 297 Germany 97
– physiological dormancy in winter 110 – gut commensals, facultative
DFD meat – See: dark, firm and dry meat pathogenic 107
direct marketing – See: local market(ing) – horizontal transmission 108
Dirofilaria – limits 19
– immitis 160 – O157 218
– repens 160 – O157:H7 231
diseases of game, South Africa 45 – venison, UK 116
disinfectant 47 EU directive (92/45/CEE) 204, 247
Distomum musculorum suis (DMS) 119 – EU ‘hygiene package’ 22, 259 – See
See also: Alaria alata also: Regulation (EC)
– adipose tissue 123 EU wild game directive 21
– detection 121, 122 evisceration 20
– digestion method 122 – infrastructure 22
– distribution 121 – in the field 83
– paratenic hosts 121 – methods 21
– predilection sites 121, 123 – partial 84, 128
– wild boar, Germany 122 – techniques 23
DMS – See: Distomum musculorum suis – time 23
dog bites 212 export
– bacteria 103, 104 – Namibia 70
– classification 102 – Namibian requirements 79, 80
dogs 101 – South Africa 56, 70
dose-response assessment 210 – South African requirements 79, 80
double sided tag 251 exposure assessment 211
drive hunts 20 exsanguination 75, 115 – See
– logistics 26 also: bleeding
extensive cattle farming 39
E
EHEC – See: Escherichia coli, F
enterohaemorhagic fallow deer, New Zealand 298
EHPs – See: Environmental Health FAO – See: Food and Agriculture
Practitioners Organisation
ehrlichiosis 170 farm abattoir 52
elk, New Zealand 298 farmed deer, UK 217
emerging pathogenic agents 159 farmed game, definition 210
emission scenarios 185, 186 farm to consumer 39
enterobacteriaceae 24, 25, 26, 27, 30, 302 feathers, microbial load 25
– venison, UK 116 field slaughter 49
Environmental Health Practitioners FMD – See: veterinary maturation of
(EHPs) 54 meat; See: foot-and-mouth disease
enzootic infections 157

Index
Food and Agriculture Organisation (FAO) – conservation 267

209 – examination by official veterinarian,
foodborne diseases 131 Austria 251
food business operator 87 – healthy product 41
food handling malpractices 219 – marbling 41
food safety – organic product 41
– criteria 31 – polyunsaturated fatty acids 41
– management 39 – resistant to spoilage 21
– management points 39, 40, 41, 60, 61 game meat examination
– training courses for direct marketing – training and evaluation concept,
262 Austria 260
Food Safety Objective (FSO) 211 Game Meat Examiner (GME) 47, 49
Food Standards Agency (FSA) 113 Game Meat Inspector (GMI) 47, 49
food supply chain, definition 41 game meat production
foot-and-mouth disease (FMD) 45, 127 – poor standardisation 31
formal risk management 209 – South Africa 68
Francisella tularensis 95, 223, 227, 236 game meat value chain 71
– Austria 161 gemsbok 68
– correlation to climate 163 Geographical Information Systems (GIS)
– coypus 236 167, 199, 200, 203
– European hares 236 Germany 19
– infection risk due to gutting and GHP – See: Good Hygiene Practice (GHP)
skinning 236 GIS – See: Geographical Information
– spatial distribution in Austria 162 Systems
– subspecies 235 GlobalGAP (Good Agricultural Practices)
– transmission to humans 235 44
free ranging reindeer 297 – elements 46
from forest/field to fork 220, 245, 259 glycolysis 278
FSA – See: Food Standards Agency GME – See: Game Meat Examiner
FSO – See: Food Safety Objective GMI – See: Game Meat Inspector
GMP – See: Good Manufacturing Practice
G good handling practices 267
game Good Hygiene Practice (GHP) 19, 30,
– consumption, Germany 94 129, 220, 245, 259, 260
– depot 49 Good Manufacturing Practice (GMP)
– handling establishments 20 104
– harvesting control document 87 greening 26
– live sales 70 green offal 52
– ranches 71 guidance values 31
game birds guides to good practice 22
– defeathering 217 gut shots 79 – See also: abdominal shot
– hung 217
game meat H
– alterations 267 HACCP – See: Hazard Analysis and
– chain, differences to slaughter 20 Critical Control Point
– cholesterol 41 haemolytic uraemic syndrome (HUS) 107

Index
haemorrhagic colitis (HC) 107 – contribution to meat supply 19

hair, microbial load 25 – definition 51
hand drying facilities 47 – historical development 93
hares, uneviscerated, storage 26 – meat 51
harvesting – motivation 51
– approved areas 81 – subsistence 51
– boma 47 – traditions 51, 247
– definition 47 HUS – See: haemolytic uraemic syndrome
– helicopter 47, 76 Hygiene Assessment System (HAS) 53
– losses 78 hygiene management system 48, 53, 129
– night time 47 H-zone 275
– of ungulates 72
– teams 47, 72, 127 I
harvesting requirements 71 ibex 157
– accustomed animals 72 – Lombardy, Italy 225
– instantaneous death 72 identification mark, game carcass 95
– minimum disturbance 72 illumination 47
– unspoilt condition 72 impala 68, 138
HAS – See: Hygiene Assessment System import, South Africa 56
haut-gout 20, 21 infectious ceratoconjunctivitis, chamois,
hazard 210 Austria 168
– biological 211 informally hunted game meat 41
– characterisation 210 inherent antimicrobial activity 137
– chemical 211 inspection 20
– physical 211 – ante and post mortem 247
Hazard Analysis and Critical Control – ante mortem 81
Points (HACCP) 55, 127, 128, 220, 262 – auxiliaries for game meat 248
– audited 217 – checklist 248
HC – See: haemorrhagic colitis – criteria, Austria 1994 248
head and (upper) neck shots 48, 73, 78 – game, Austria 1994 249, 250
health certificates, large game 256 – large wild game, Germany 95
heat-shortening 282 – modular scheme 247
heavy metals 212 – post mortem 49, 216
hepatitis E 95, 170 – pre-harvesting 48
– Germany 97 – risk-based 246
hide harvesting 75 – three-step scheme 248
human health, fatty acids intramuscular connective tissue 274
– omega-3/omega-6 ratio 309 ISO standard 6579:2002 133
– PUFA 309 Istituto Zooprofilattico Sperimentale della
– SFA 309 Lombardia e dell’Emilia-Romagna
human larval alariosis 120 (IZSLER) 225
hunters 246, 248 Italian Alps 267
– food business operators 22 IZSLER – See: Istituto Zooprofilattico
– non-professional 22 Sperimentale della Lombardia e
hunting dell’Emilia-Romagna
– biltong 51

Index
K – pH 289, 290, 291

Krebs cycle 276 – pheasants 291, 292
Krim-Kongo-Fever 170 – physico-chemical 273, 289
kudu 68 – roe deer 289
– seasonal variation 289
L – sensory 273, 289
lactate 277 – shear force 289, 290, 291
lactic acid 326 – standard methods of assessment 315
– change during storage 329 – water holding capacity 289, 290, 291
– pheasant, breast muscles 328, 329 – wild boar 291
– pheasant, thigh muscles 329 meat temperatures, max (EU) 23
large game hunting, Portugal 101 mercury 212
large-scale processors 55 microbial
large wild game, skinning 217 – contamination, visual cleanliness 27
lead 87, 212 – numbers 26
– pellets 216 – transfer skin/hair to meat 25
– shot 218 microbiology
Leishmania infantum 160 – condition 102
Listeria monocytogenes 264, 269 – EU criteria 24
local market(ing) 93, 94, 259, 260 – limits 32, 263
– legal framework, Austria 261 – limits, carcasses 28
– Lower Austria 262 – limits, fresh meat 31
– structure, Lower Austria 264 – own-checks 263
local trade – See: local market(ing) – pH 31
louping ill 170 – quality, venison, UK 114
lures 256 – relation to sensory characteristics 31
– wild ruminants, Belluno, Italy 270
M – wild ungulates, Belluno, Italy 269
meat damage microflora
– due to bullet wound 78 – game bird 30
– due to dogbite 101 – meat cuts 28, 29
meat flavour – meat from small game, GHP
– factors affecting 287 conditions 30
– grassy, gamey and wild 308 – roe deer meat 30
meat pH – surface of wild game carcasses 27
– glycogen content 302 M-line 275
– high, bacterial spoilage 71 mobile slaughter facilities 300
– high, muscle (DFD) 77 monitoring
– microbiological effects 302 – and surveillance, OIE 236
– quality indicator 301 – for residues 50
– tenderness 302 MtbC – See: Mycobacterium tuberculosis
– ultimate pH, reindeer 305 complex
meat quality muscle
– ante and post mortem factors 273 – biological mechanisms 273
– chamois 290 – colour determiation factors 319
– concepts 273 – glycogen stores 301

Index
– proteins, classification 274 permanent food premises, checklist 262

muscle fibre types 274 pH
– red/oxidative 306 – change during storage 328
– white/glycolytic 306 – measurement in muscle 326
Mycobacterium – measurement in watery suspensions
– avium 218 326
– bovis 218, 223, 227, 233 – pheasant, breast muscles 328, 329
– bovis, in wildlife, Italy 234 – pheasant, thigh muscles 328, 329
– microti, in wildlife, Italy 234 – post mortem muscle 315
– tuberculosis complex (MtbC) 233 pheasants
myofibrillar proteins – ammonia 325
– actin 274 – lactic acid 325
– arrangement 274 – pH 325
– myosin 274 – post mortem changes 325
myoglobin, relation to oxygen pressure – uneviscerated 217
279 – uneviscerated, storage 26
myosin 275 – unviscerated, shelf-life 24
placement of shots 22
N PO – See: Performance Objective
Namibia 67, 70, 127 poly-unsaturated fatty acids (PUFA) 308
– wildlife numbers 69 population, critical size 167
National Reference Centre for Wildlife Portugal 101
Diseases (Ce.R.M.A.S.), Italy 224 post-hunt presentation (‘Streckenlegung’)
New Zealand, venison production system 22
298 post mortem tissues 25
Niedere Tauern 168 potable water 47
night harvesting 72 pre-cooling 24, 27
– ante mortem stress 73 primary production 21
nyala 138 – level 33
product quality, ISO definition 315
O professional cooling facilities 24
offal handling 50 proteolysis 283
official controls, UK, prior to 2006 217 PSE meat – See: pale, soft and exudative/
ostrich 138 watery meat
own-checks 246, 259 PUFA – See: poly-unsaturated fatty acids
P Q
pale, soft and exudative meat (PSE meat) Q fever 159, 160
71, 279, 284, 286 qualitative risk assessment, hazards, wild
park deer, slaughter 115 game, UK 218
passive sanitary surveillance quality meat programmes 31
– Belluno, Italy 267 questionnaire 39
– vademecum 267
PC – See: Performance Criterion R
Performance Criterion (PC) 211 radioactive fallout 212
Performance Objective (PO) 19, 211 red deer, New Zealand 298

Index
refrigeration 22 – serotypes in wildlife, Italy 231

Regulation (EC) – See also: EU ‘hygiene – venison, UK 115
package’ – wild boars, Italy 231
– No. 178/2002 95, 252, 260 – wild boars, Portugal 132
– No. 852/2004 252, 260, 262 Salmonella Rissen 133
– No. 853/2004 93, 252, 260 Salmonella Typhimurium 133
– No. 854/2004 252 – wild boars 134
– No. 2073/2005 263 Sami reindeer 299
– No. 2075/2005 227, 253 – Fennoscandia 297
reindeer husbandry 310 sample size, problems with small animals
reticulo-endothelial system 139 323
rigor contracture 282 sanitary surveillance 267
rigor mortis 278, 326 Sanitation Standard Operating Procedure
rinderpest 127 (SSOP’s) 128
risk 210 sarcomere length 282
– actual 215 – measurement by laser diffraction 322
– definition (FAO/WHO) 213 saturated fatty acids (SFA) 308
– perceived 215 SEIR model – See: susceptible, exposed,
risk analysis 209, 210 infectious, recovered model
– definition (FAO/WHO) 210 self assessment 53
risk assessment 210 self-checks – See: own-checks
– policy 210 sensory panel studies 322
risk characterisation 210 serious abnormalities 249
risk communication 210 SFA – See: saturated fatty acids
risk estimate 210 shiga toxin producing Escherichia coli
risk management 209, 210 (STEC) 95, 107
– Codex Alimentarius Commission – Germany 97
principles 213 skeletal muscle, composition 274
– decisions 214, 216 skin, microbial load 25
– framework 214 snow grouse 157
– monitoring and review 214, 216 SOP’S – See: Standard Operating
– options 214, 215 Procedures
– preliminary activities 214 South Africa 39
– preliminary tasks 215 South African
risk pathways 218 – legislation, large game 79
risk profile 210 – wildlife industry 44
rural community development 40 springbok 68
SSOP’s – See: Sanitation Standard
S Operating Procedure
Salmonella 95, 131, 212, 218, 223, 227, Standard Operating Procedures (SOP’S)
264, 269 128
– detection method 115, 133 Staphylococcus aureus 25, 116, 137, 138
– in wildlife 132 STEC – See: Shiga toxin producing
– red deer, Italy 231 Escherichia coli
– red foxes, Italy 231 stifling maturation/stickige Reifung 23
– roe deer, Italy 231 still hunting 20, 247

Index
supply chain 39 – Italy 230

– South Africa 42, 43 – life cycle 230
surplus game animals 70 – reducing risks 230
survey, Lower Austria 262 traceability 39, 95, 251
susceptible, exposed, infectious, recovered – feedstuff 256
model (SEIR model) 175 – tagging of carcasses and pluck 50
suspect carcass form 83 tradition 20, 22
sustainable – hunting, South Africa 46
– development 40 trained persons 94, 246, 248
– extensive conservation 40 – education, Austria 250
– utilisation 39 – harmonisation in EU 256
sustained harvest 71 training 33, 52, 246
Swedish reindeer herding 300 – books 21
– for hunters 52, 94
T – for local marketing 263
TAC – See: Total Aerobic Counts – of harvesters 81
taste panel 322 transport
temporary – methods, South Africa 58
– food premises 262 – to field abattoir 83
– slaughter depots 47 Trichinella 21, 95, 223, 227, 249
tenderness – accreditation of laboratories 99
– collagen isoforms 281 – accredited laboratory 254
– connective tissue 281 – advanced training for inspectors 255
– deer species 306 – basic training for inspectors 254
– factors affecting 284, 320 – britovi 227
– measurement 320 – britovi, Austria 252
– MIRINZ tenderometer 321 – compression method 97
– muscle preparation for measurement – digestion method 97, 253, 254
321 – domestic cycle 144, 155, 226
– post mortem tenderisation 283 – epidemiology, Austria 252
– Volodkevich device 321 – genotypes 144
– Warner-Bratzler device 321 – infective dose 146
thermograph 47 – infectivity 145
thoracic shots 82 – inspection 252
throat slitting 82 – life cycle 145
thrombotic-thrombocytopenic purpura – monitoring in wildlife 227
(TTP) 107 – proficiency test 254, 255
time between killing and processing 216 – pseudospiralis 227, 253
time/temperature profile from killing to – red fox, Italy 227
cooling 27 – red fox, reservoir, Austria 252
Total Aerobic Counts (TAC) 24, 26, 27, – self-evaluation of trained persons 255
30 – See also: Aerobic Colony Counts – species 143
(ACC) – species in Europe 227
– limits 19 – sylvatic cycle 144, 155, 226
Toxoplasma gondii 223, 227 – training of hunters for sampling 97
– in wildlife 230 – wild boars, Italy 227

Index
– wild game, Germany 96 U

Trichinella control unapproved food chain 216
– domestic pigs 153 Usutu virus (USUV) 170
– home-made pork products 154 – blackbirds 174, 175
– post-harvest 154 – block diagram 176
– principles 153 – dynamics 189
– wildlife 153 – endemicity, Central Europe 190
Trichinella-free status 154, 227 – epidemic model 174, 176
trichinellosis – model, bird parameters 177
– European Union 147 – model, equations 195
– heat inactivation 151, 152 – model, mosquito parameters 179
– home-made pork products 152 – model, simulation 1901-2100 188
– sensitive to freezing 151, 152 – model, simulation 2001-2005 182
– Serbian perspective 143 – model, source-code (R) 197
– source 151 – model, transmission parameters 178
– (wild) pigs, Serbia 147, 148 – mosquitoes (Culex pipiens) 174, 175
trichinellosis, humans – Vienna, Austria 173
– intestinal phase 146 USUV – See: Usutu virus
– muscular phase 146
– Serbia 149 V
– source attribution, Serbia 152 vector-borne diseases 159
– therapy 146 venison
trichinoscopy 253 – definition 40
– proficiency test 254, 255 – ethics 310
– restrictions, Austria 254 – impact of growth rate 306
– trained persons 254, 255 – metabolism, photoperiod-induced
trophy hunting 41, 51, 70 changes 299
troponin complex 275 – myoglobin 307
TTP – See: thrombotic-thrombocytopenic – positive image 310
purpura – sources of deer, UK 115
tuberculosis 45 ventilation 24
– fallow deer 199 verocytotoxic Escherichia coli (VTEC)
– Iberian Peninsula 200 107, 223, 227
– Idanha-a-Nova, Portugal 201 – ante mortem condition 109
– infection pathways 233 – contamination pathways 109
– in wildlife, Italy 233 – epidemiology 231
– Lombardy 233 – farm animals 107
– mouflon 199 – herbal foodstuffs 108
– red deer 199 – O26:H11 108
– wild boars 199, 203 – O103:H2 108
tuberculosis-like lesions 233 – O128:H2 108
– wild boar lymph nodes 234 – post mortem conditions 110
tularaemia 157 – See also: Francisella – roe deer, Italy 232
tularensis – wild ruminants, Germany 108, 109
– Germany 98 verotoxins (shigatoxins) 107
– hares 98 veterinary health plan 44, 46

Index
veterinary maturation of meat (FMD) 85, – monitoring of diseases 225, 227

128 – monitoring of diseases, Italy 224, 236
– CCP 129 – negative perception 39
vitamin E 307 – population, Lombardy and Emilia-
VTEC – See: verocytotoxic Escherichia coli Romagna, Italy 226
– quantity, Austria 246
W – ranching 40
wapiti, New Zealand 298 – source of human disease 224
warmed-over flavour 288 winter feeding 256
warthog 138 wolf 225
waste handling 50 World Health Organisation (WHO) 209
water-binding capacity – See: water- wounding pattern 20
holding capacity (WHC)
water-holding Y
– fluid accumulation 285 Yersinia
– myofibrils shrinkage 284 – enterocolitica 223, 227, 232
– osmotic pressure 285 – in wildlife, Italy 233
– protein denaturation 285 – pseudotuberculosis 218
water-holding capacity (WHC) 316 – species, Italy 233
– cooking loss 317
– drip loss 317 Z
– heating loss 317 zebra 68, 138
– methods for assessment 316 zoonotic agents 95, 223
– press-method 316
West Nile virus 170
– Austria 159
WHC – See: water-holding capacity
WHO – See: World Health Organisation
wild animals – See: wildlife
wild boars
– direct marketing 253
– faecal shedder of Salmonella 133
– inspection, Austria 253
– trichinoscopic method 253
wildebeest 68, 138
Wildfleischanhänger 251
wild game – See: wildlife
wildlife
– conservancies 71
– cycling of infection 131
– dangers to domestic animals 44
– definition 210
– health 224
– industry 70
– Lombardy and Emilia-Romagna, Italy
225

Game Meat Hygiene in Focus Microbiology Epidemiolo

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Game meat hygiene in focus

Game meat hygiene in focus

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Having regard of the above-mentioned considerations, a consortium of food hygienists has

Game meat hygiene in focus 7

Vienna and Brno, February 2011

Section 1 – Hygiene and microbiology

Hygiene and microbiology of meat from wild game: an Austrian view 19

Zoonotic diseases and direct marketing of game meat: aspects of consumer

Game meat hygiene in focus 9

Verocytotoxigenic Escherichia coli (VTEC) in wild ruminants in Germany 107

Microbial quality of venison meat at retail in the UK in relation to

Detection of Alaria spp. mesocercariae in game meat in Germany 119

10 Game meat hygiene in focus

Preliminary results indicating game meat is more resistant to

Game meat hygiene in focus 11

Dynamics of infectious diseases according to climate change: the Usutu virus

The utility of GIS in studying the distribution of Bovine Tuberculosis in wild

12 Game meat hygiene in focus

Risk Management of game: from theory to practice 209

The monitoring of selected zoonotic diseases of wildlife in Lombardy and

Game meat hygiene in focus 13

Approaches to game hygiene in the province Belluno (Italy): from training to

Section 4 – Muscle biology and meat quality

The muscle biological background of meat quality including that of

Muscle biological and biochemical ramifications of farmed game husbandry

14 Game meat hygiene in focus

A summary of methods to assess major physical-chemical and sensory

Evaluation of some parameters of post mortem changes of pheasant

Biographies of key contributors 333

Game meat hygiene in focus 15

P. Paulsen et al. (eds.), Game meat hygiene in focus, 19

20 Game meat hygiene in focus

Game meat hygiene in focus 21

In summary, improvements in hygiene of the slaughter of farm animals and in cooling

22 Game meat hygiene in focus

Table 1. Time from killing to evisceration, large game (% of carcasses).

<1 h 1-3 h >3 h n= Reference

17.5 75.7 5.8 103 Riemer and Reuter, 1979

Game meat hygiene in focus 23

A ‘pre-cooling’ phase, as generally recommended for carcasses from slaughter animals in

1.3 Panta rhei

2. Microbiology of meat from wild game

24 Game meat hygiene in focus

2.1 Microflora of live game/uneviscerated carcass

Muscle tissue of healthy slaughter animals is generally considered as virtually sterile

2.2 Contamination at primary production level (shooting, evisceration and

Game meat hygiene in focus 25

26 Game meat hygiene in focus

2.2.3 Surface microflora on wild game carcasses in the first 24 h p.m.

2.2.4 Microbial contamination and visual assessment of ‘cleanliness’ of carcasses

Game meat hygiene in focus 27

2.2.5 Microbial limits for carcasses of wild game?

2.3 Microflora of meat cuts

2.3.1 Microflora of meat cuts: surveys and experimental findings

TAC Enterobacteriaceae E. coli Based on

6.0 - 2.0 Deutz et al., 2003 (non destructive sampling)

28 Game meat hygiene in focus

Sampling n TAC (log cfu/g) EB (log cfu/g or cm 2) Sensory assessment Reference