Iranian Journal of Veterinary Research, University of Shiraz, Vol. 7, No. 2, Ser. No.

15, 2006

Aerobic bacteria isolated from eggs and day-old

chicks and their antibacterial resistance
in Shiraz, Iran
Nazer, A. H. K.1*; Dadras, H.2 and Eskandari, S.1, 3
1
Department of Food Hygiene and Public Health, School of Veterinary Medicine, University of Shiraz, Shiraz,
Iran; 2Department of Avian Medicine, School of Veterinary Medicine, University of Shiraz, Shiraz, Iran; 3Food
and Drug Control Laboratories, Ministry of Health and Medical Education, Tehran, Iran
*
Correspondence: A. H. K. Nazer, Department of Food Hygiene and Public Health, School of Veterinary
Medicine, University of Shiraz, Shiraz, Iran. E-mail: nazer@shirazu.ac.ir

Summary
To study the putative transfer of antibiotic resistance from broiler breeders to human, hen’s eggs and their
day-old chicks were examined for the presence of bacteria. The most frequently isolated organisms in
decreasing order were: Streptococcus spp., Bacillus spp., Staphylococcus spp., Klebsiella spp., Enterobacter
spp. and Escherichia coli followed by Citrobacter spp., Proteus spp. and Pseudomonas spp. from the eggs and
E. coli, Enterobacter spp. and Citrobacter spp. followed by Klebsiella spp. and Bacillus spp. from the chicks.
Different detection methods were evaluated which use various enrichment and plating media for bacteria in
eggs and day-old chicks. Sensitivity tests showed the presence of antibacterial resistant strains of bacteria. In
comparison, resistance to all antibiotics in chicks’ isolated bacteria were more frequent than eggs’ isolates, but
statistically no significant differences between patterns of antibacterial resistance were seen (P ≤ 0.05).
Twenty-three, 54, 55, 60, 24 and 10% of chicks’ isolates were resistant to chloramphenicol, enrofloxacin,
erythromycin, furazolidone, trimethoprim and tylosin, respectively. Whereas these data about eggs’ isolates
were as follows: 1, 12, 18, 18, 10 and 6%, respectively. This study revealed that eggs are often contaminated
with different bacteria and could be potential vehicles for transmitting of these bacteria through their broilers.
Our findings stress the need for increased implementation of hazard analysis of critical control points
(HACCP) and consumer food safety education efforts.

Key words: Eggs, Day-old chicks, Antibacterial resistance, Public health

Introduction impair the quality of chicks produced. In the

case of table eggs spoilage may occur and if
Microbial food safety is an increasing public the organism is of public health significance,
health concern worldwide. Data recorded in the affected eggs may be the cause of spread
different countries have shown that the of disease. The majority of the salmonellosis
incidence of some of food-borne diseases outbreaks is associated with consumption of
have increased dramatically over the past few eggs and egg dishes. There are two ways in
years, but because of under-responding, the which eggs can become contaminated,
data are of limited value and can not be namely, by the transovarian and trans-shell
compared between countries (Sackey et al., routes. In transovarian contamination the egg
2001; Zhao et al., 2001). becomes contaminated prior to oviposition,
The relationship of the hen’s egg to human with the source of contamination originating
and animal health depends to some extent on in the egg-laying apparatus of the bird. In the
its microbial content, more especially on the case of trans-shell contamination, the
microbiology of the freshly laid egg. organisms gain access to the egg after
Microbial contamination of eggs is a oviposition by penetration into the shell.
well-established phenomenon and has These organisms could be derived from
important economic implications to the either the intestinal tract or the environment.
poultry industry. Contamination of hatching Contact with contaminating organisms when
eggs may reduce hatchability, be responsible the egg shell is wet may also facilitate the
for transmission of poultry pathogens and penetration of the pathogens. When eggs are

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Iranian Journal of Veterinary Research, University of Shiraz, Vol. 7, No. 2, Ser. No. 15, 2006

broken, bacteria present on egg shells may month for 8 months. A total number of 114
contaminate the contents. These eggs and 120 day-old chicks were examined,
con-taminants may grow rapidly in broken from a broiler breeder company in Shiraz
out egg if storage is at ambient temperature area. Each egg sample was aseptically
(Board and Fuller, 1994; Grijspeerdt, 2001; removed and placed in a plastic container and
Hara-Kudo et al., 2001; Radkowski, 2001). transferred to the lab. The chicks (Aryan
Eggs are produced by laying hens (layers); hybrid) were kept in special boxes at the
however, common approaches are applied in laboratory, have been brought there
the supply of layers. As nearly all birds are immediately after hatching. Fertile eggs were
derived from a very small number of hatched in the company’s own hatchery.
elite/great-grandparent supply points, it is
clear that any pathogens entering the Breaking of eggs for culturing
population in these early stages will be able The egg-shell was wiped with a sterile
to spread throughout the layer populations cotton wool swab moistened with sterilized
very quickly indeed (Bell and Kyriakides, normal saline (0.85%), then wiped with a
2002). Among the available methods for the cotton ball soaked in 70% ethanol and finally
control of these pathogens, the one most it was sterilized by a quick passing over a
widely practiced is the use of various flame. This procedure was followed to avoid
antibiotics, fungicides and coccidiostats in contamination of the egg contents from the
the birds’ diet. Nevertheless, it is well known germs colonizing the egg-shell
that the extended and continuing use of a (Papadopoulou et al., 1997;
range of antimicrobial agents in animals’ Himathon-gkham et al., 1999).
food has been an important factor in
promoting the emergence of resistant strains Preparing the chicks for
of Gram-positive and Gram-negative bacteria bacterio-logical examination
(Papadopoulou et al., 1997; Aarestrup et al., Killing the day-old chicks was performed
2000). Resistant organisms can spread from following disarticulating cervical vertebrate
chicken to chicken and from chicken to man method recommended by Strafuss (1988).
(Levy et al., 1976). In 1992, an article that Necropsy procedure was preformed
was published in Science focused on following the method recommended by
antimicrobial resistance and listed the “top Strafuss (1988) and immediately after death
ten drug-resistant microbes”: one-half were the abdomen was quickly opened and the
Gram-negative bacteria, including whole intestine removed and unraveled with
Enterobacteriaceae, Haemophilus influenzae, sterile precautions (Smith, 1965).
Neisseria gono-rrhoeae, Pseudomonas
aeruginosa and Shigella dysenteriae. Culturing method
Although resistant Gram-positive bacteria, After disinfection, each egg was cracked
particularly enter-ococci, pneumococci and with a sterile surgical knife and its content
staphylococci are clearly a problem, resistant (white and yolk) was dropped into glass
Gram-negative bacteria remain an important container containing 150 ml of trypton soya
cause of morbidity and mortality. The cost of broth (TSB, Merck) (Papadopoulou et al.,
treatment is also a problem for infections 1997; Himathongkham et al., 1999). The
caused by Gram-negative organisms intestinal samples were squeezed into a tube
(Gibbons, 1992). containing 9 ml of TSB (Barnes et al., 1972).
For the purpose of studying antibacterial After homogenization, the TSB cultures were
resistance, potentially transmitted from incubated at 37ºC for 18-24 hrs and then
poultry to humans, hens’ eggs and their subcultured to suitable selective media.
day-old chicks were examined for the These media were McConkey agar (Merck)
presence of bacteria. The eggs and chicks for the cultivation of Enterobacteriaceae,
used came from a broiler breeder farm in double concentration selenite enrichment
Shiraz area. broth (Merck) as enrichment broth and
Materials and Methods brilliant green phenol red lactose agar
(Merck) for Salmonella isolation.
Sample collection Mueller-Hinton agar (Merck) and blood agar
Sampling visits were made in every other

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Iranian Journal of Veterinary Research, University of Shiraz, Vol. 7, No. 2, Ser. No. 15, 2006

(Oxoid) were used for the cultivation of other formaldehyde-DMF (furazolidone)

Gram-positive bacteria (Papadopoulou et al., (Andrews, 2001).
1997; Himathon-gkham et al., 1999).
Selective enrichment for Salmonella was Methods to test susceptibility
carried out using 225 ml of double The susceptibility tests were performed
concentration selenite enrichment broth for following the dilution antimicrobial tests
the content of each egg, or 25 ml liquid egg recommended by Quinn et al., (1994) and
and 10 ml for intestinal contents and Hirsh and Zee (1999). This test was
incubated at 37°C for 24 hrs under aerobic performed by preparing two-fold dilutions of
conditions. Enrichment was carried out for an antibiotic in a series of tubes containing
all samples (Hara-Kudo et al., 2001; Sackey Mueller-Hinton broth. From each bacterial
et al., 2001). Selenite enrichment broth was strain an inoculum previously adjusted to 0.5
then streaked onto McConkey agar and Unit of the McFarland scale and then diluted
brilliant green phenol red lactose agar as 1 : 100 to obtain 104 and 105 bacteria/ml
indirect plating. Escherichia coli and other concentration. Each tube was inoculated with
Enterobacteriaceae were isolated using a suspension of the test bacterium. The
McConkey agar and eosin methylene blue inoculated tubes of broth were incubated at
agar (BioMérieux). Incubation was done at 35-37°C for 16-18 hrs. The highest dilution
37°C for 24-48 hrs (Sackey et al., 2001). of the antibiotic to inhibit visible growth of
After incubation an isolated colony was bacterium (no turbidity in the tube) was used
picked from a suitable plate and subcultured as the minimum inhibitory concentration
into another plate to obtain a pure culture. (MIC). To determine the resistant isolated
The identification of the isolated bacteria was bacteria, following breakpoints of antibiotics
based on standard bacteriological and were considered: chloramphenicol ≥ 3200
biochemical procedures. Strains µg/ml (Aarestrup et al., 2000; White et al.,
characte-rized by Gram stain and hemolysis 2003); enrofloxacin ≥ 400 µg/ml (White et
on sheep blood agar. The Gram-positive al., 2000); erythromycin ≥ 800 µg/ml
cocci were first classified upon their reaction (Aarestrup et al., 2000; White et al., 2003);
to the catalase test and then further identified furazolidone ≥ 200 µg/ml (National
using the oxidase test and other biochemical Committee for Clinical Laboratory
reactions. The Gram-negative bacteria were Standards, NCCLS, Guidelines, Chicago
first classified upon their reaction to the Department of Public Health, 1998);
oxidase test and then further identified by trimethoprim ≥ 1600 µg/ml (Aarestrup et
using the biochemical tests (Clarke and al., 2000) and tylosin ≥ 3200 µg/ml (White
Bauchop, 1977; Quinn et al., 1994; Mahon et al., 2003).
and Manuselis, 1995; Papadopoulou et al.,
1997; Himathongkham et al., 1999). Statistical analysis
Data were analysed using the SPSS statistic
Antimicrobial susceptibility testing software version 11.5. Independent t-test was
used to find the significant differences
Antimicrobials between the two groups (P ≤ 0.05).
A total of 6 antibacterials were used as the
following: trimethoprim, tylosin, Results
erythromycin and enrofloxacin (approved
Various Gram-positive and Gram-negative
drugs in Iran) and chloramphenicol and
bacteria were isolated. Thirty-three (28.94%)
furazolidone (not approved for use in animal
out of 114 examined eggs were contaminated
in Iran).
with 46 bacteria (9 species) that 27.3% of
Antimicrobial powders were obtained from
these eggs were contaminated with more than
different companies in Iran. Firstly, the purity
one bacterium. The rate of microbial
of them was measured with diffusion method
contamination of eggs with E. coli in this
(Brooks et al., 1998) and then, antimicrobial
study was 8.7% (n = 4). Our study indicated
stock solutions were prepared and stored in
the following rates for contamination with
95% ethanol solution (chloramphenicol and
other Entero-bacteriaceae: Klebsiella spp.
erythromycin), distilled water (trimethoprim,
15.22% (n = 7), Proteus spp. 2.17% (n = 1),
tylosin and enrofloxacin) and dimethyl

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Iranian Journal of Veterinary Research, University of Shiraz, Vol. 7, No. 2, Ser. No. 15, 2006

Enterobacter spp. 10.87% (n = 5) and Enterobacter spp. (12.5%) and E. coli

Citrobacter spp. 2.17% (n = 1). Also one (20.4%). 18.75% of Citrobacter spp.,
Pseudomonas spp. (2.17%) was detected. 48.97% of E. coli and 12.5% of Enterobacter
Gram-positive bacteria had these rates of spp. isolated from chicks were resistant to
contamination: 21.74% Streptococcus spp. (n enrofloxacin. In the isolated bacteria from
= 10), 17.4% Staphylococcus spp. (n = 8) and chicks, resistance to erythromycin was seen
19.56% Bacillus spp. (n = 9). in 55.1 and 6.25% of E. coli and Citrobacter
Ninety (75%) out of 120 tested day-old spp., respectively. All bacteria isolated from
chicks were contaminated with 144 bacteria chicks showed resistance to furazolidone as
(5 species) that 55.56% of these chicks were the following percentages: Bacillus spp.
contaminated with more than one bacterium. (100%), Citrobacter spp. (18.75%),
In this case, E. coli had the most incidence Enterobacter spp. (29.16%), E. coli (48.97%)
(68.05%) followed by Enterobacter spp. and Klebsiella spp. (20%). 21.42, 8.34 and
(16.67%), Citrobacter spp. (11.11%), 6.25% of E. coli, Enterobacter spp. and
Klebsiella spp. (3.47%) and Bacillus spp. Citrobacter spp. isolated from chicks were
(0.7%). At last, the resistance of all isolated resistant to trimethoprim, respectively.
bacteria against several antibiotics Resistance to tylosin had the least level in
commonly used in chicken industry in Iran chicks’ isolated bacteria: 9.2% of E. coli and
was examined. The antibiotic susceptibility 4.16% of Enterobacter spp.
of isolates to the different antibiotics is A very frequently occurrence of resistance to
shown in Tables 1 and 2. tested antibiotics was observed among both
Ten percent of egg isolates (n = 1) were groups, but statistically no significant
resistant to chloramphenicol. This resistance differences between the pattern of
was pertaining to Streptococcus spp. No antibacterial resistance were seen (P ≤ 0.05).
resistance was seen in other isolates. 55.56,
25, 14.28, 25 and 30% of Bacillus spp., E. Discussion
coli, Klebsiella spp., Staphylococcus spp. and
Streptococcus spp. isolates from eggs were The present study was conducted to
resistant to enrofloxacin, respectively. determine the species distribution and
Among resistance to erythromycin in susceptibility to antimicrobial agents among
bacteria isolated from eggs, these results bacteria isolated from eggs and day-old
were obtained: Bacillus spp. (55.56%), E. chicks in Shiraz, Iran. The use and misuse of
coli (75%), Klebsiella spp. (14.28%), antibiotics contribute to the development of
Staphylococcus spp. (37.5%) and resistance and it is generally in agreement
Streptococcus spp. (60%). that this is a function of the span of time and
With exception of Proteus spp. and use; therefore, it is of basic importance to
Pseudomonas spp., all isolated bacteria from implement monitoring systems. A common
eggs were resistant to furazolidone in the limitation of monitoring systems is that they
following order: Citrobacter spp. (100%), usually consider the resistance only to
Bacillus spp. (66.67%), Klebsiella spp. antimicrobial drugs of clinical isolates. In
(42.85%), Streptococcus spp. (40%), E. coli view of the expected correlation between
(25%), Staphylococcus spp. (25%) and animal food and human clinical disease, we
Enterobacter spp. (20%). Resistance to decided to direct our investigation towards
trimethoprim was seen in 11.14% of Bacillus the strains isolated from food instead of
spp., 20% of Enterobacter spp., 25% of E. clinical isolates. In this way, we could
coli, 28.58% of Klebsiella spp., 25% of establish an Iranian database to be
Staphylococcus spp. and 30% of periodically updated to foresee the trend of
Streptococcus spp. isolates from eggs. bacterial resistance to antibiotics. Thus,
11.14% of Bacillus spp., 25% of E. coli, selected strains of bacteria were recovered
14.28% of Klebsiella spp., 12.5% of from eggs and day-old chicks. Also an
Staphylococcus spp. and 20% of assessment of the incidence and type of
Streptococcus spp. isolates from eggs were bacterial contamination occurring in eggs
resistant to tylosin. produced and hatched in commercial
In the isolated bacteria from day-old chicks, hatcheries is essential for understanding the
resistance to chloramphenicol was limited to role that microorganisms play in influencing

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hatchability. Pseudomonas spp., all of other isolated

The results of the present study indicate the bacteria from eggs were resistant to this
existence of a variety of bacteria in the egg antibiotic in high percentage. This high
which can transfer to human via different resistance to furazolidone also was seen in all
foods or their chicks. It was also observed isolated bacteria from chicks (Table 2).
that of all eggs examined (114), 28.94% were Natural contamination of egg contents with
found to be contaminated with more than one bacteria such as Salmonella and E. coli can
organism with the combination of occur in two ways. One is penetration from
Enterobacteriaceae and Streptococcus spp. outside the egg into the content. Another
occurring more frequently than would be mode of natural contamination occurs in the
expected by chance. This can be explained by reproductive tract, probably the upper
assuming that the organism originates from a oviduct, with the most important sites for
common source, namely faeces. It should contamination being the outside of the
also be noted that the various pathogens were vitelline membrane and the surrounding
isolated from egg-yolk after sterilization of albumen (Humphrey and Whitehead, 1993;
the shell egg to minimize contamination from Humphrey, 1994). It was suspected that the
germs colonizing the egg shells. contamination was a result of incomplete
In this study, the bacterial flora recorded disinfection of the shell and/or membrane
from the eggs showed that the predominant resulting in transfer of bacteria to the content
species were Streptococcus spp., Bacillus during the egg breaking procedure.
spp., Staphylococcus spp. and Klebsiella spp. The normal gut flora of chicks is highly
comprised 21.73, 19.56, 17.39 and 15.21% of complex and not yet fully understood. A
the total bacteria isolates, respectively and number of naturally occurring and artificial
other groups present lower levels. All these factors are able to affect the composition of
bacteria (Bacillus spp., Citrobacter spp., the flora. These factors include age, the
Enterobacter spp., E. coli, Klebsiella spp., immune response, diet and orally
Proteus spp., Pseudomonas spp., administered antibiotics (Board and Fuller,
Staphylococcus spp. and Streptococcus spp.) 1994). Although the alimentary tract of the
have been isolated from eggs in other studies healthy newly hatched chick is usually sterile
and our results are in general agreement with it rapidly becomes colonized by facultative
the results obtained by Taku et al., (1986), anaerobic bacteria, particularly coliforms and
Board and Fuller (1994), Papadopoulou et streptococci (Board and Fuller, 1994). Work
al., (1997) and Zhao et al., (2001). by Fuller and Jayne-Williams (1968)
Moreover, the bacteria were isolated from demonstrated bacterial contamination of the
eggs coming from large industry-scale peritoneal cavity, 38% and yolk sac
broiler breeder plant, where antibiotics are infections, 23% in 121 conventional chicks
not widely used. Taking into consideration examined during the first 5 days of life.
that furazolidone had used for the control of Considerable variation, related to incubator
infections before laying period for a week hygiene was observed between different
and comparing the resistance of the isolated batches of chicks. The more frequently
bacteria to this specific antibiotic, it is quite isolated organisms were streptococci
possible that resistant bacteria could be followed by, in decreasing order, micrococci
passed to human through the food chain and coliform organisms (mainly E. coli). The
(Papadopoulou et al., 1997). As showed in micrococci were considered to be the
Table 1, except of Proteus spp. and characteristic of the

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Table 1: Antimicrobial susceptibility of isolated bacteria from eggs in Shiraz, Iran

Antimicrobial Bacterial Number of isolates with MIC (µg/ml) Resistance
agent species (%)
6.25 ≥ 12.5 25 50 100 200 400 800 1600 ≥ 3200
Bacillus spp. 1 1 2 2 1 1 - 1 - - 0
Citrobacter - - - - - 1 - - - - 0
Enterobacter - 2 - 1 - 1 - - 1 - 0
Escherichia 1 - 1 1 - - - 1 - - 0
Chloramphenicol * Klebsiella - 2 - 1 1 2 1 - - - 0
Proteus - - - 1 - - - - - - 0
Pseudomonas - 1 - - - - - - - - 0
Staphylococcus - 1 - 1 2 3 - 1 - - 0
Streptococcus - 1 1 1 2 1 2 1 - 1 10

Bacillus spp. 1 1 - 1 - 1 2 2 1 - 55.56

Citrobacter 1 - - - - - - - - - 0
Enterobacter 2 1 - - 1 1 - - - - 0
Escherichia 1 - 1 - - 1 - 1 - - 25
Klebsiella 2 - - 1 1 2 1 - - - 14.28
Proteus - - 1 - - - - - - - 0
Pseudomonas - 1 - - - - - - - - 0
Staphylococcus 1 2 1 - 1 1 1 - 1 - 25
Streptococcus 2 1 1 2 1 - 2 - 1 - 30

Bacillus spp. - - 1 - 2 - 1 3 1 1 55.56

Citrobacter - - - 1 - - - - - - 0
Enterobacter 1 - - 2 - 1 1 - - - 0
Escherichia - - 1 - - - - 1 - 2 75
Erythromycin * Klebsiella 1 1 - - 1 1 2 - 1 - 14.28
Proteus - - - - 1 - - - - - 0
Pseudomonas - - - - - 1 - - - - 0
Staphylococcus 1 - - 2 - 1 1 - 1 2 37.5
Streptococcus - - - - 1 2 1 1 3 2 60

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Table 1 (Cont): Antimicrobial susceptibility of isolated bacteria from eggs in Shiraz, Iran
Antimicrobial Bacterial Number of isolates with MIC (µg/ml) Resistance
agent species (%)
6.25 ≥ 12.5 25 50 100 200 400 800 1600 ≥ 3200
Bacillus spp. - 2 1 - - 1 2 1 1 1 66.67
Citrobacter - - - - - - 1 - - - 100
Enterobacter 1 - - 2 1 - 1 - - - 20
Escherichia 1 - - 1 1 - - - 1 - 25
Furazolidone *** Klebsiella - 1 - 1 2 - 1 1 - 1 42.85
Proteus - - - - 1 - - - - - 0
Pseudomonas - - - - 1 - - - - - 0
Staphylococcus 1 1 - 1 3 - - - 1 1 25
Streptococcus 1 1 - 3 1 1 - 2 - 1 40

Bacillus spp. - 2 1 1 2 1 - 1 1 - 11.14

Citrobacter - - - 1 - - - - - - 0

Enterobacter - 1 - - 1 1 - 1 - 1 20
Escherichia - - 1 - - 1 - 1 - 1 25
Klebsiella - - - 1 2 1 1 - 2 - 28.58
Proteus - - - - - - - 1 - - 0
Pseudomonas - - - - 1 - - - - - 0
Staphylococcus 1 - 1 - 1 1 2 - - 2 25
Streptococcus - 1 1 1 - 2 1 1 2 1 30

Bacillus spp. 1 1 - 1 2 1 - 2 - 1 11.14

Citrobacter - - - - 1 - - - - - 0
Enterobacter - 1 1 - 1 1 - 1 - - 0
Escherichia - - 1 - - 1 1 - - 1 25
Tylosin ***** Klebsiella 2 - 2 1 - - - 1 - 1 14.28
Proteus - - - - 1 - - - - - 0
Pseudomonas - - - - - - 1 - - - 0
Staphylococcus 1 - 2 - 2 1 1 - - 1 12.5
Streptococcus 1 - 2 - 1 1 2 - 1 2 20

Bold lines indicate breakpoints for resistance according to: *Aarestrup et al., (2000) and White et al., (2003); **White et al., (2000); ***National Committee for Clinical
Laboratory Standards (NCCLS) Guidelines (Chicago Department of Public Health, 1998); ****Aarestrup et al., (2000); *****White et al., (2003)

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Table 2: Antimicrobial susceptibility of isolated bacteria from day-old chicks in Shiraz, Iran
Antimicrobial Bacterial Number of isolates with MIC (µg/ml) Resistance
agent species (%)
6.25 ≥ 12.5 25 50 100 200 400 800 1600 ≥ 3200
Bacillus spp. - - 1 - - - - - - - 0
Citrobacter 2 2 1 1 1 6 1 2 - - 0
Chloramphenicol * Enterobacter 2 7 2 1 2 3 2 1 1 3 12.5
Escherichia 12 6 9 14 10 5 3 8 11 20 20.4
Klebsiella - 2 1 1 - 1 - - - - 0
Bacillus spp. - - - - - 1 - - - - 0
Citrobacter 6 2 1 3 1 - 1 2 - - 18.75
Enrofloxacin ** Enterobacter 8 1 3 2 5 2 1 2 - - 12.5
Escherichia 7 5 14 8 3 13 7 14 9 18 48.97
Klebsiella 2 1 - 1 - 1 - - - - 0
Bacillus spp. - - 1 - - - - - - - 0
Citrobacter 1 3 1 1 5 2 2 1 - - 6.25

Enterobacter 2 4 3 7 2 4 2 - - - 0
Escherichia 4 12 5 7 9 3 4 17 11 26 55.1
Klebsiella - 2 - 1 - 1 1 - - - 0
Bacillus spp. - - - - - 1 - - - - 100
Citrobacter 2 1 3 4 3 1 2 - - - 18.75
Furazolidone *** Enterobacter 4 5 - 3 5 3 2 1 1 - 29.16
Escherichia 17 14 5 10 4 14 11 2 9 12 48.97
Klebsiella 1 1 - - 2 - 1 - - - 20
Bacillus spp. - 1 - - - - - - - - 0
Citrobacter 2 2 1 3 2 1 2 2 1 - 6.25
Trimethoprim**** Enterobacter 1 4 1 2 5 3 2 4 1 1 8.34
Escherichia 8 10 19 5 4 11 6 14 6 15 21.42
Klebsiella 1 - - 2 1 - 1 - - - 0
Bacillus spp. - - - - 1 - - - - - 0
Citrobacter 2 3 1 2 3 1 2 1 1 - 0
Tylosin ***** Enterobacter 2 6 2 3 2 4 - 3 1 1 4.16
Escherichia 16 8 9 5 13 6 15 10 7 9 9.2
Klebsiella 1 1 2 - 1 - - - - - 0

Bold lines indicate breakpoints for resistance according to: *Aarestrup et al., (2000) and White et al., (2003); **White et al., (2000); ***National Committee for Clinical
Laboratory Standards (NCCLS) Guidelines (Chicago Department of Public Health, 1998); ****Aarestrup et al., (2000); *****White et al., (2003)

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Fig. 1: The number of isolated resistant bacteria from eggs and day-old chicks to the used antibiotics

chick’s gut prior to feeding, but in our study the finding of Board and Fuller (1994) that
no micrococci were seen. Facultative the coliforms are the only organisms
anaerobes include members of normally present in the chicks gut flora. In
Entero-bacteriaceae such as E. coli, the study of Nazer and Safari (1994), isolates
Citrobacter spp., Proteus spp. and Klebsiella of bacteria comprising E. coli (37.64%),
spp. which are frequently present but in Klebsiella spp. (14.11%), Bacillus spp.
lower numbers. Smaller numbers of other (2.35%) were cultured from dead-in shell
organisms such as the aerobe, Pseudomonas chick. In our study E. coli was found in larger
spp. and yeasts may be found throughout the numbers throughout the alimentary tract of
gut from time to time (Clarke and Bauchop, chicks (68.04%) that has similar range of
1977; Board and Fuller, 1994). Our results bacterium found by Smith, (1965) and
were in general agreement with mentioned Rajaian et al., (2002). Other major
surveys. differences between isolated bacteria from
Since faecal contamination is thought to two groups are that the eggs had an incidence
be a major cause of egg contamination by of Proteus spp., Pseudomonas spp.,
Salmonella, it is not surprising that other Staphylococcus spp. and Streptococcus spp.,
members of the Enterobacteriaceae, while the chicks did not have them. These
particularly E. coli, can also be isolated from results indicate poor nest hygiene which
eggs. Between 0.5 and 6% of eggs from could have provided an opportunity for
normal hens contain E. coli. Thus, hatched contamination the eggs with fecal organisms.
chicks may already have E. coli-infected yolk Also under the conditions employed for
sacs leading to neonatal mortality. However, incubating the eggs, coliforms would be
other organisms including Proteus spp. and preferentially selected in favour of other
enterococci may also be involved, suggesting microorganisms. The absence of any
involvement of the gut flora (Board and Salmonella among both groups has to be
Fuller, 1994). Our results showed that about noted in this study.
3.5% of eggs were infected with E. coli, but The sensitivity test performed showed
81.65% of chicks showed contamination with the presence of resistant bacteria (Fig. 1). The
this micro-organism. finding of chloramphenicol resistance in eggs
The results obtained in this paper confirm (2.17%) and chicks (15.97%) was not

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Iranian Journal of Veterinary Research, University of Shiraz, Vol. 7, No. 2, Ser. No. 15, 2006

expected, because this antibiotic must not be from poultry. Vet. Microbiol., 74: 353-364.
used in poultry production in Iran. Also this 2- Andrews, JM (2001). Determination of
resistance pattern to other antibiotics (except minimum inhibitory concentrations. J.
of furazolidone) that have not been used in Antimicrob. Chemother., (Suppl. S1), 48:
5-16.
tested farm was seen. Illegally use of this
3- Barnes, EM; Mead, GC; Barnum, DA and
antibiotic in poultry farms and transmission Harry, EG (1972). The intestinal flora of the
of resistant bacteria via wild birds, workers, chicken in the period 2 to 6 weeks of age, with
equipments and also through feed could be particular reference to the anaerobic bacteria.
the reason. The occurrence of enrofloxacin Br. Poult. Sci., 13: 311-326.
resistance among chicks’ isolated bacteria 4- Bell, C and Kyriakides, A (2002). Salmonella:
was higher (37.5%) than that observed a practical approach to the organism and its
among the same bacteria of eggs (26.08%). control in foods. 7th. Edn., Blackwell Science.
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