Nature and Science, 2012;10(3) http://www.sciencepub.

net/nature

Microorganisms Associated With Spoilage Of Stored Vegetables In Uyo Metropolis, Akwa Ibom State,
Nigeria

Adebayo-Tayo BC1, Odu NN2, Osunnwoke UJ1, Okonko IO2

1
Department of Microbiology, Faculty of Science, University of Ibadan, Ibadan, Nigeria
2
Department of Microbiology, University of Port Harcourt, Choba, P.M.B. 5323, Port Harcourt, River State,
Nigeria;
3
Department of Microbiology, Faculty of Science, University of Uyo, Uyo, Akwa Ibom State, Nigeria
mac2finney@yahoo.com, iheanyi.okonko@uniport.edu.ng

Abstract: Microorganisms associated with spoilage of stored vegetables were studied using standard
microbiological methods. The analysis was done on carrots, cucumber, cabbage and onions with soft rot symptoms
using various media. It showed that Escherichia coli (28.6%) were the most predominant bacterial isolates
associated with vegetable spoilage in Uyo metropolis. This was followed by Enterobacter spp. (21.4%),
Staphylococcus aureus (14.3%), Erwinia spp. (14.3%) and Pseudomonas spp. (14.3%) while Salmonella spp.
(7.1%) was least predominant. The results obtained from the microorganism s associated with the spoilage of stored
vegetables showed that the total aerobic counts for cucumber ranged from 1.28 x 10 6 to 3.20 x 106 cfu/g and the total
coliform count ranged from 2.35 x 10 6 to 3.28 x 106 cfu/g. The total aerobic count for carrots ranged from 2.07 x 10 6
to 2.20 x 106 cfu/g while the total coliform count ranged from 2.80 x 10 6 to 3.00 x 106 cfu/g. The total aerobic count
for cabbage ranged from 1.43 x 106 to 2.10 x 106 cfu/g while the total coliform count ranged from 3.10 x 10 6 to 4.20
x 106cfu/g. Generally, the total Salmonella-Shigella counts ranged from 0.0 x 10 6 to 1.80 x 106 cfu/g. Only cabbage
had the highest count for total Salmonella-Shigella. The aerobic count for onions ranged from 1.8 x 10 6 to 2.0 x 106
cfu/g and the total coliform count ranged from 2.00 x 10 6 to 3.10 x 106 cfu/g. Fungi isolated from these vegetables
were Aspergillus niger, Rhizopus stolonifer and Aspergillus fumigatus with fungal count ranging from 1.80 x 10 6 to
3.0 x 106 cfu/g. The frequency of occurrences showed that bacterial isolates were most predominant (63.6%)
compared to the fungi isolates (36.4%). No protozoa or viruses were found to be associated with vegetable spoilage
in Uyo metropolis. The frequency of occurrences of bacteria isolates showed that Escherichia coli (28.6%) were
most. This was followed by Enterobacter spp. (21.4%), Staphylococcus aureus (14.3%), Erwinia spp. (14.3%) and
Pseudomonas spp. (14.3%). Salmonella spp. (7.1%) was least predominant. It also showed that of the three fungi
species isolated from vegetables, Rhizopus stolonifer (37.5%) and Aspergillus fumigatus (37.5%) were most
predominant while Aspergillus niger (25.0%) was least predominant. Pathogencity tests revealed that all the isolates
were pathogenic to the different vegetables examined leading to rapid disintegration of treated fruits in 3-5 days. It
showed that each infected vegetable gave the initial organism that caused the spoilage of the fruit. The rot symptoms
obtained were similar to those observed previously on the fruits when subjected to identification procedures. The
presence of indicator and other organisms examined in this study is of special concern and perhaps the greatest
danger associated with food for human consumption is contamination by human excrement. The need for microbial
assessment of vegetables for production of salads and other use is also emphasized to reduce possible contamination.
[Adebayo-Tayo BC, Odu NN, Osunnwoke UJ1, Okonko IO. Microorganisms Associated With Spoilage Of Stored
Vegetables In Uyo Metropolis, Akwa Ibom State, Nigeria. Nature and Science 2012;10(3):23-32]. (ISSN: 1545-
0740). http://www.sciencepub.net. 4

Keywords: Food spoilage, index of food quality, pathogens, Vegetables

1. Introduction proper function of the body. Vegetable contain various

Generally, vegetables are considered as the medicinal and therapeutic agent and are valued mainly
leafy outgrowth of plants or plants shoot used food. for their high vitamin and mineral content. Studies
These include those plants or plant part used in making have evaluated the association of fruit and vegetable s
soup or served as an integral part of main meal (Yusuf consumption with the reduction of risk of specific
et al 2004). Vegetables can also be regarded as the diseases (Hung et al., 2004).
edible component of plants, such components includes The incidence of microorganisms in
leaves, stalk, roots, tubers, bulbs, flowers and seed vegetables may be expected to reflect the sanitary
(ICMSF 1998). Vegetables are important protective quality of the processing steps and the microbiological
food and highly beneficial for the maintenance of condition of the raw product at the time of processing
health and prevention of diseases. They contain (Ngugen and Carlin, 1994). For almost 100 years,
valuable food ingredients which are essential for the vegetables contaminated in the field have been

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recognized as a source of human infection and unless. enzymes producing free fatty acids and glycerol
Many of the viruses (Rosenblum et al., 1990), bacteria (Pomeranz 1992; Bulaong and Dharmaputra, 2002).
(Ho et al., 1986) and protozoan on vegetables which Altogether, these changes lead to a lower quality or
have caused food poisoning are derived from human rejection of commodity as foodstuff (Bulaong and
faeces. However, pathogenic microorganism of human Dharmaputra, 2002). Three fungal species, namely
origin may also be present in minimally processed Aspergillus flavus, A. parasiticus, and A. nomius
vegetables as the minimal technological processing produce aflatoxins as secondary metabolites (Pitt and
may be unable to remove the original contamination Hocking 1997). The toxins are known to be
resulting from air, soil, water, insects, animals, carcinogenic, hepatotoxic and teratogenic in test
workers, harvesting and transportation equipment. animals (Bulaong and Dharmaputra, 2002). Moisture
Certain fungi such as Aspergillus, Fusarium, and content is the most important factor affecting fungal
Penicillium spp. as commonly occurring filamentous growth in stored products (Bulaong and Dharmaputra,
fungi grow in vegetable and their growth may result in 2002). According to Sauer et al. (1992) some fungal
production of toxins known as mycotoxins, which can species were observed to become dominant with as
cause a variety of ill effect in human from allergic little as 0.2% change in moisture content.
responses to immunosuppression and cancer (Pitt et al., Spoilage microorganisms can be introduced
1996). to the crop on the seed itself, during crop growth in the
The ability of public health agencies to field, during harvesting and postharvest handling, or
identify through enhanced epidemiological and during storage and distribution (Barth et al., 2009).
surveillance techniques, raw vegetables as probable Those same types of soil-borne spoilage microbes that
sources of infectious microorganisms has undoubtedly occur on produce are the same spoilage
resulted in increased numbers of documented microorganisms that are present on harvesting
outbreaks. The risk of illness associated with raw equipment, on handling equipment in the
vegetable products can be reduced by removing or packinghouse, in the storage facility, and on food
killing pathogenic microorganisms by washing or contact surfaces throughout the distribution chain
treating them with sanitizers. However, the (Barth et al., 2009). Therefore, early intervention
hydrophobic cutin, diverse surface morphologies and measures during crop development and harvesting
abrasions in the epidermis of fruits and vegetables through the use of good agricultural practices (GAP)
limits the efficacy of this treatment (Burnett and will provide dramatic reductions in yield loss due to
Beuchat, 2001). spoilage at all subsequent steps in the food-to-fork
Vegetables are frequently consumed raw continuum (Eckert and Ogawa, 1988; Barth et al.,
without being exposed to the processes that reliably 2009).
eliminates pathogens. Washing fruits and vegetables in The problem of fungal growth and aflatoxin
chlorinated water can reduce bacterial levels but cannot contamination of foodstuffs remains, especially in
be relied upon to eliminate pathogens. Eating or developing countries where handling and storage
drinking contaminated foods or drinks can cause food- technologies are still being developed (Bulaong and
borne disease. Many different types of bacteria, viruses Dharmaputra, 2002). Pitt and Hocking (1996) reported
and parasites can contaminate food, so there is that aflatoxin exceeding 50 ppb contaminated 45, 22
numerous different food borne infections. The and 25% of 215, 81 and 94 peanut samples collected
consumption of carrot, cucumber, onions and cabbage from retailers in 1990/1991 in Indonesia, Philippines
in Nigeria has increased tremendously in the recent and Thailand, respectively. In line with this study
years properly due to increased awareness on their Lubulwa and Davis (1994) reported that the total
health important. Carrot is known to contain an annual cost of aflatoxin contamination in Indonesia,
important biologically active compound, carotenoid Philippines and Thailand was about $A 158 million.
(Asagbra and Oyewole, 2002). It has been estimated Indonesia incurred 84% (= $A 132 million) of this
that 20% of vegetables harvested for human cost, Thailand incurred 13% (= $A 21 million) and
consumption are lost through microbial spoilage by one Philippines 3% (= $A 5 million) of the cost.
or more of 250 market diseases, the primary causative Dharmaputra and Putri (1997) found that P.
agents of microbial spoilage are the bacteria, yeasts and funiculosum was a dominant species in peanuts with
mold (Jay, 2005). different percentages of splitted kernels (0, 25, 50, 75
Storage fungi can cause decrease of and 100%) stored for 2 months under laboratory
germination capability, loss in weight, discoloration of conditions. Its population was between 0.7x10 -
kernels, heating and mustiness, chemical and 1.2xl03 cfu/g (wb). Pitt et al. (1998) reported that P.
nutritional changes, and mycotoxin contamination funiculosum were found in 4% of 256 retail samples
(Sauer et al. 1992; Bulaong and Dharmaputra, 2002). examined and 9% of all kernels examined (50
They can change fat quality of peanuts by hydrolytic kernels/sample) in Bogor and Yogyakarta, Indonesia.

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It is estimated that about 20-25% of the harvested 1972). For bacterial colony purification, the streak
fruits are decayed by pathogens during post-harvest techniques in the Nutrient agar was employed, while
handling even in developed countries (Droby, 2006; the fungal isolates were transplanted to new set
Zhu, 2006; Al-Hindi et al., 2011). Sabourand agar by picking. The bacterial plates were
However, in Nigeria, local utilization of incubated at 370C for 24 h. The fungal plates were left
carrots, cabbage, onions, and cucumbers is limited to at room temperature for 5 days. The discrete colonies
direct unprocessed eating either wholly or salads. This from these subcultured plates and series of
situation has lead to a growing awareness on the need biochemical tests were done on the bacterial isolates
to evaluate microorganism associated with spoilage of for proper characterization and identification. The
these vegetables sold in Uyo metropolis. The aim of bacterial isolates were also identified by comparing
this wok was to determine the microbial load of stored their characteristics with those of known taxa, as
vegetables sold at direction locations in Uyo, and to described by Jolt et al. (1994) and Oyeleke and Manga
isolate, characterized and identify the specific (2008). The pure isolated fungi were identified using
organisms in different stored vegetables in Uyo cultural and morphological features according to the
metropolis, Akwa Ibom State, Nigeria. most documented keys in fungal identification
(Domsch et al., 1993; Klich, 2002; Samson and Varga,
2. Materials And Methods 2007).
2.1. Collection of samples
Twenty four (12 healthy and 12 infected) 2.3. Pathogenicity of isolated organisms
samples each of carrot, cabbage, cucumber and onions Twelve (12) healthy vegetables collected from
were purchased from the market place and storage the each study sites were surfaced sterilized in ethyl
room all around Uyo metropolis, Akwa Ibom State, alcohol as described above. Vegetables were wounded
Nigeria. These samples were placed in separate sterile with a sterile 5 mm cork borer, and inoculated with
plastic bags and transported to the laboratory for bacterial colony and mycelia disc (3 mm in diameter)
microbial analysis. These vegetables were not locally of the fungal test isolate. The inoculated wound was
cultivated in the state. A total of 12 randomly selected sealed with Vaseline petroleum jelly. The inoculation
infected vegetables and 12 unblemished, healthy and was done in a laminar flow chamber. Twelve
clean looking vegetables were purchased. Vegetables vegetables were inoculated each with each of the
were surface sterilized by exposing them in 1 min 90% isolates and this experiment was replicated. Controls
ethyl alcohol (BDH chemicals Ltd Poole England) and consisted of twelve vegetables wounded with the
then 3 min to 1% sodium hypochlorite and then rinsed sterilized cork borer but not inoculated. The inoculated
three times in sterile distilled water. Segments (3 - 5 fruits and the controls were placed in clean
cm) of tissues from the margins of the rotted areas were polyethylene bag (one fruit per bag) each moistened
cut out with a sterile scalpel and placed on previously with wet balls of absorbent cotton wool to create a
prepared media in Petri dishes and incubated at humid environment and incubated at 30 + 1OC and
appropriate temperatures. 37OC for 5 days. After 72 h, the inoculated vegetables
were observed for symptom development. The causal
2.2. Enumeration, Isolation and Identification of agents were re-isolated from the infected vegetables
Microorganisms and compared with the original isolates.
Samples were stored and allowed to ferment.
The spoiled parts of each samples from different 3. RESULTS ANALYSIS
location was weighed and grinded using mortar and 3.1. Total Bacteria and Fungi Counts Obtained
pestle. One gram of each sample was dispensed into a from Vegetables
prepared 9 ml of distilled water contained in the The results obtained from the microorganism s
McCartney bottles. The content was shaken for associated with the spoilage of stored vegetables
homogenous mixture. Ten fold serial dilutions were showed that the total aerobic counts for cucumber
made and 10-5 dilution of the samples from different ranged from 1.28 x 106 to 3.20 x 106 cfu/g and the total
location were plated out using Nutrient agar, coliform count ranged from 2.35 x 106 to 3.28 x 106
MacConkey and Salmonella-Shigella agar for total cfu/g. The total aerobic count for carrots ranged from
aerobic and coliform counts. These were incubated for 2.07 x 106 to 2.20 x 106 cfu/g while the total coliform
24 hours at 370c in the incubator. Sabourand dextrose count ranged from 2.80 x 106 to 3.00 x 106 cfu/g. The
agar and potato dextrose agar (PDA, Difco) were used total aerobic count for cabbage ranged from 1.43 x 106
for the total fungal counts and incubated at 28 ± 1oC to 2.10 x 106 cfu/g while the total coliform count
for 5 days under 12 h photoperiod. The fungal colonies ranged from 3.10 x 106 to 4.20 x 106cfu/g. Generally,
that appeared were primarily identified using cultural the total Salmonella-Shigella counts ranged from 0.0 x
and morphological features (Barnett and Hunter, 106 to 1.80 x 106 cfu/g. Only cabbage had the highest

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count for total Salmonella-Shigella. The aerobic count x 106 cfu/g. The total fungal count ranged from 1.80 x
6 6
for onions ranged from 1.8 x 10 to 2.0 x 10 cfu/g and 106 to 3.00 x 106 cfu/g (Table 1).
6
the total coliform count ranged from 2.00 x 10 to 3.10
Table 1: Total Bacteria and Fungi Counts Obtained from Vegetables
Samples Location Total Aerobic Total Coliform Total Salmonella- Total Fungal
Counts (CFU/g) Counts (CFU/g) Shigella Counts (CFU/g) Counts (CFU/g)
Cucumbe Storage room 1.28 x 106 2.35 x 106 0.0 x106 1.80 x 106
r
Market place 3.20 x 106 3.48 x 106 0.0 x106 2.00 x 106
Carrot Storage room 2.07 x 106 2.10 x 106 0.0 x106 2.10 x 106
Market place 2.20 x 106 3.0 x 106 0.0 x106 2.50 x 106
Cabbage Storage room 1.43 x 106 3.10x 106 1.30 x106 2.70 x 106
Market place 2.10 x 106 4.20 x 106 1.80 x106 2.07 x 106
Onions Storage room 1.80 x 106 2.20 x 106 0.0 x106 2.80 x 106
Market place 2.80 x 106 3.00 x 106 0.0 x106 3.00 x 106

3.2. Frequency of occurrences of Microorganisms Uyo metropolis. It showed that bacterial isolates were
Associated with Vegetables spoilage in Uyo most predominant (63.6%) compared to the fungi
Metropolis isolates (36.4%). No protozoa or viruses were found to
Table 2 shows the frequency of occurrences of be associated with vegetable spoilage in Uyo
microorganisms associated with vegetables spoilage in metropolis (Table 2).

Frequency of occurrences of Microorganisms Associated with Vegetables spoilage in

Table 2:
Uyo Metropolis.
Isolates No. (%) Cucumber Carrot Cabbage Onions
Bacteria 14(63.6) 3(21.4) 4(28.6) 4(28.6) 3(21.4)
Fungi 8(36.4) 3(37.5) 2(25.0) 2(25.0) 1(12.5)
Protozoa 0(0.0) 0(0.0) 0(0.0) 0(0.0) 0(0.0)
Viruses 0(0.0) 0(0.0) 0(0.0) 0(0.0) 0(0.0)
Total 22(100.0) 6(27.3) 6(27.3) 6(27.3) 4(18.2)

3.3. Frequency of occurrences of Bacteria and Pseudomonas spp. (14.3%). Salmonella spp.
Associated with Vegetables spoilage in Uyo (7.1%) was least predominant (Table 3). Escherichia
Metropolis coli were present in all the vegetables examined.
Table 3 shows the frequency of occurrences of Enterobacter spp. was present in all the vegetables
bacteria associated with vegetables spoilage in Uyo examined except for cabbage, in which it was not
metropolis. It showed that Escherichia coli (28.6%) found. Salmonella spp. was only present in cabbage.
were most predominant bacterial isolates associated Staphylococcus aureus was only found in cucumber
with vegetable spoilage in Uyo metropolis. This was and carrot. Erwinia spp. was present in carrot and
followed by Enterobacter spp. (21.4%), cabbage only. Pseudomonas spp. was present in
Staphylococcus aureus (14.3%), Erwinia spp. (14.3%) cabbage and onions only (Table 3).

Table 3:Frequency of occurrences of Bacteria Associated with Vegetables spoilage in Uyo

Metropolis.
Bacteria No. (%) Cucumber (%) Carrot (%) Cabbage (%) Onions (%)
Escherichia coli 4(28.6) 1(25.0) 1(25.0) 1(25.0) 1(25.0)
Pseudomonas spp. 2(14.3) 0(0.0) 0(0.0) 1(50.0) 1(50.0)
Erwinia spp. 2(14.3) 0(0.0) 1(50.0) 1(50.0) 0(0.0)
Staphy. aureus 2(14.3) 1(50.0) 1(50.0) 0(0.0) 0(0.0)
Enterobacter spp. 3(21.4) 1(33.3) 1(33.3) 0(0.0) 1(33.4)
Salmonella spp. 1(7.1) 0(0.0) 0(0.0) 1(100.0) 0(0.0)
Total 14(100.0) 3(21.4) 4(28.6) 4(28.6) 3(21.4)

3.4. Frequency of occurrences of Fungi Associated and Aspergillus fumigatus (37.5%) were most
with Vegetables spoilage in Uyo Metropolis predominant while Aspergillus niger (25.0%) was the
Table 4 shows the frequency of occurrences of least predominant (Table 4). Aspergillus niger was
fungi associated with vegetables spoilage in Uyo present in both cucumber and onions only. Rhizopus
metropolis. It showed that of the three fungi species stolonifer and Aspergillus fumigatus were present in all
isolated from vegetables, Rhizopus stolonifer (37.5%) vegetables except for onions (Table 4).

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Table 4: Frequency of occurrences of Fungi Associated with Vegetables spoilage in Uyo Metropolis.
Fungi No. (%) Cucumber (%) Carrot (%) Cabbage (%) Onions (%)
A. niger 2(25.0) 1(50.0) 0(0.0) 0(0.0) 1(50.0)
A.fumigatus 3(37.5) 1(33.3) 1(33.3) 1(33.4) 0(0.0)
R. stolonifer 3(37.5) 1(33.3) 1(33.3) 1(33.4) 0(0.0)
Total 8(100.0) 3(37.5) 2(25.0) 2(25.0) 1(12.5)
4. DISCUSSION al., 2005). In this present study, all the vegetables
The total aerobic counts obtained in this study examined haboured Escherichia coli, while prominent
is lower compared to that reported by Kaneko et al. microorganisms variously haboured include
(2003), however, the total coliform counts and the total Enterobacter spp., Staphylococcus aureus, Erwinia
fungal counts were slightly higher than that reported by spp., Pseudomonas spp., Salmonella spp., Aspergillus
Kaneko et al. (2003). The bacteria isolates obtained in niger, Rhizopus stolonifer and Aspergillus fumigatus.
this study were also slightly different from those The detection of E. coli in this study showed poor
identified by Kaneko et al. (2003) in a similar study. hygienic standard in the handling of these salad
The initial bacteria of stored produce may have been vegetables or it could be also be from contamination
derived from contamination of air, soil, water, insects, during harvest. Presence of E. coli indicates recent
animals, workers and harvesting and transportation contamination by faecal matter and possible presence
equipments (FDA, 2002). . Also, the densities of Lactic of other enteric pathogens known to be causative
acid bacteria (LAB) in fruit and vegetable products agents of food borne gastroenteritis and bacterial
usually range from 102 to 106 CFU/wound (Trias et al., diarrhea disease (Jiwa et al., 1981).
2008). In the study by Trias et al. (2008), the highest Most strains of Staphylococcus aureus are
concentrations of microorganisms were in ready-to-eat known to be pathogenic due mostly to the heat stable
vegetables. This was due to the presence of cut enterotoxin they produce in direct relationship to their
surfaces, which allow higher nutrient availability inoculum level. Considering the notoriety of the
(Ongeng et al., 2006) and affects not only LAB but all resistance of S. aureus to methicillin, other penicillin
the microbiota related to the fresh product (Badosa et and celphalosporins (Davis, 1997; Adeleke and
al., 2008; Trias et al., 2008). The microbial population Odelola, 1997), its detection in cucumber and carrot
levels found in this study were in agreement with data samples poses a lot of health risk to nourishment
reported for ready-to-eat salads in other studies (Carlin seeking consumers. The presence of Staphylococcus
et al., 1989; Trias et al., 2008). auerus, a pathogenic organism of public health concern
The bacterial isolates identified in this study and significance in these vegetables might have
include Escherichia coli, Enterobacter spp., contaminated the stored vegetables from source as a
Staphylococcus aureus, Erwinia spp., Pseudomonas result of handling by farmers or retailers. Improper
spp., and Salmonella spp. This is consistent with the handling and improper hygiene might lead to the
findings of previous studies. Microorganisms most contamination of food and this might eventually affects
commonly found in vegetables generally involve the health of the consumers (Dunn et al., 1995; Omemu
Pseudomonas and Erwinia as coliforms and and Bankole, 2005; Okonko et al., 2008 a,b,c,d,
Micrococcus (ICSMF, 1998). Sufficient moisture, 2009a,b; Mgbakor et al., 2011).
abusive temperature and adequate time will ensure a Most of the organisms found in this study are
continuing increase in the bacteria population. those commonly found in soil and water. But the
Pathogens such as Bacillus cereus, Salmonella and presence of other indicator organisms like E. coli and
Escherichia coli are naturally present in some soil, and Enterobacter aerogenes in those vegetable samples
their presence on fresh produce is not rare. The genus might be the result of possible contamination during
Erwinia belongs to the family Enterobacteriacease. sales or unhygienic handling of stored vegetables. The
They are all associated with plants where they are presence of the most frequently isolated index of food
known to cause plant diseases of the rot and wilt type. quality and indicators of faecal contamination such as
These Gram negative rods that are related to the genera Escherichia coli Enterobacter aerogenes and
Proteus, Serratia, Escherichia, Salmonella and others. Salmonella spp., reported in this study is an indication
E. coli and Salmonella spp. are indicators of feacally of faecal contamination of the food as a result of
contaminated products or vegetables. possible unhygienic handling (Okonko et al., 2008
Food processors may be sources of microbial a,b,c,d 2009a,b) or contamination of the vegetable
chance inoculation, microbial food poison, food itself during processing or directly from source and this
intoxication and food spoilage hence, food processors might have adverse effect on the health of the
may be counter productive by being responsible for consumers (Okonko et al., 2008a,b,c,d, 2009a,b).
public health hazard and loss of revenue (Bankole et

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In this study, the fungi isolated were 1983). Aflatoxin production depends among others on
Rhizopus stolonifer, Aspergillus fumigatus and the strain of Aspergillus sp. (Kozakiewicz 1996; Pitt
Aspergillus niger. These pathogens have been and Hocking 1997; Bulaong and Dharmaputra, 2002).
reportedly isolated from Pawpaw fruits in Nigeria Generally, spoiling fungi are considered
(Baiyewu et al., 2007; Chukwuka et al., 2010; toxigenic or pathogenic (Al-Hindi et al., 2011).
Akintobi et al., 2011). Several fruit spoilage fungi Toxigenic fungi have been isolated from spoiling fruits
from different region has been isolated and identified (Stinson et al., 1981; Al-Hindi et al., 2011). During
(Al-Hindi et al., 2011). The most common fungi refrigeration some moulds may produce mycotoxins
found in a study by Akintobi et al. (2011) were (Tournas and Stack, 2001; Al-Hindi et al., 2011).
Aspergillus flavus, A. niger, Fusarium solani, Pathogenic fungi, on the other hand, could cause
Penicillium digitatum, Rhizopus stolonifer and yeasts. infections or allergies (Monso, 2004; Al-Hindi et al.,
Baiyewu et al. (2007) had also reported that Rhizopus 2011). Aspergillus spp. are known to produce several
nigricans, Aspergillus flavus, Aspergillus niger and toxic metabolites, such as malformins, naphthopyrones
Fusarium moniliforme among others, were responsible (Frisvad and Samson, 1991; Pitt and Hocking, 1997;
for post harvest losses in Pawpaw in South Western Al-Hindi et al., 2011) and they can produce
Nigeria. Chukwuka et al. (2010) recently reported that Ochratoxins (OTA), a mycotoxin which is a very
R. nigricans, A. flavus, A. niger, Fusarium sp., and important toxin worldwide because of the hazard it
Mucor sp. were responsible for with Pawpaw fruits poses to human and animal health (Peraica et al., 1999;
decay from a farm in Oyo State, South Western Petzinger and Weidenbach, 2002) thus extra care
Nigeria. The isolation of Rhizopus stolonifer from should be taken during personnel handling of these
vegetables further confirmed the studies of fruits; such as harvesting, cleaning, sorting, packaging,
Efiuvwerwere (2000), Chuku et al. (2008), transport and storage (Al-Hindi et al., 2011).
Akinmusire (2011), and Akintobi et al. (2011) who All the three fungi organisms isolated were
reported that Fusarium sp, and Rhizopus stolonifer confirmed to be pathogenic on vegetables but in
were responsible for the soft rot of tomato. Onyia et varying degrees. It showed that all the isolated fungi
al. (2005) also reported that Fusarium monilifome, were highly pathogenic leading to rapid disintegration
Aspergillus niger and Rhizopus stolonifer were of treated vegetables in 3-5 days. These three fungi
isolated from rotten tomato fruits. isolates (Rhizopus stolonifer, Aspergillus fumigatus
Aspergillus spp. were widespread among all and Aspergillus niger) were successfully taking part in
examined vegetables spoilage. A. niger is a fungus the vegetable decay and are thus confirmed as the
commonly found on grapes (Chulze, 2006), apples causal organisms of decay (Baiyewu et al., 2007;
(Oelofse, 2006) and tomatoes (Yildz and Baysal, Chukwuka et al., 2010; Akintobi et al., 2011). Okereke
2006). Bali et al. (2008) reported that black mold A. et al. (2010) indicated that the fungi species isolated
niger were caused post harvest spoilage in sweet from the infected mangoes included A. niger,
orange and acid lime at field. The group of diseases Fusarium sp and A. Flavus and that Fusarium sp and
caused by Aspergillus are called aspergillosis, the A. Flavus could not prove pathogenicity when
symptoms include fever, cough, chest pain or inoculated into healthy mango fruits.
breastlessness. Usually, only patients with already The contamination of vegetables by
weakened immume systems or who suffer other lung pathogenic bacteria and fungi could also be as a result
condition are susceptible. Gupta and Pathak (1986) of poor handling practices in food supply chain,
had earlier reported that Aspergillus niger, Aspergillus storage conditions, distribution, marketing practices
flavus, Rhizopus nigrican, Curvularia lunata, and transportation (Effiuvwevwere, 2000; Okonko et
Rhizopus oryzae, Fusarium equiseti and Fusarium al., 2008a,b,c,d, 2009a,b; Akinmusire, 2011; Akintobi
moniliforme were responsible for post harvest losses et al., 2011). Post harvest handling and transport of
in pawpaw in South Western Nigeria. Krogh (1992) fruit is inadequate (Baiyewu et al., 2007). Therefore
has earlier reported that most microbes infecting plant most of the fruits and vegetables harvested do not
tissues often produced secondary metabolites in their usually get to the major cities in time due to the nature
hosts, which are known to be hazardous to animals of transport systems existing in the rural areas
including man. Some of these metabolites include the (Baiyewu et al., 2007; Akintobi et al., 2011). While
ergot alkaloids on cereals by Clavisep sp, fumonisin fruit with bruises are not isolated from the unbruised
on maize by Fusarium sp, aflatoxins and ochratoxins ones and thereby causing cross-infections, consumers
on several plants produced by Aspergillus sp. (Prasad, are supplied mostly with partly rotten fruits (Baiyewu
1992). Aflatoxins, which are a group of highly toxic, et al., 2007). This portends a great risk of aflatoxin and
mutagenic and carcinogenic polyketide compounds, other mycotoxins to the consumers (Baiyewu et al.,
were first reported in groundnut feed that poisoned 2007; Akintobi et al., 2011).
thousands of poultry and pigs (Goldblatt and Stoloff,

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 Nature and Science, 2012;10(3) http://www.sciencepub.net/nature

5. CONCLUSION practices can result in contamination as well as poor

Microorganisms are naturally present on all handling by dealers, processing and consumer in the
foodstuffs and can also be brought in by outside home. Side by side is the huge nutritional benefit
elements (wind, soil, water, insects, animals, human derivable from consumption of these vegetable
handling. They can become contaminated during especially that are therapeutic, curative and preventive.
growing, harvesting and transport of the raw materials, Just as with other foods consumer have some
and/or processing into finished products (Lelieveld et responsibilities to carry when handling these
al., 2003). It is therefore necessary and important that vegetables, wash hand with warm water and soap
both the farmer who harvests the fruits into bags for before and after handling the vegetables. Rinse raw
transportation, the marketers and consumers take product in clean water, brush off debris. For cutting,
necessary and appropriate precautions in preventing plastic board should be used instead of wooden ones.
contamination and eating of contaminated fruits
(Baiyewu et al., 2007). This will however reduce the CORRESPONDENCE TO:
risk of mycotoxins associated with fungi Iheanyi O. Okonko
contamination which are deleterious to human health Department of Microbiology,
(Baiyewu et al., 2007; Chukwuka et al., 2010). University of Port Harcourt,
From this study, some of the isolated East/West Road, P.M.B 5323, Choba,
organisms have serious public health risk while others Port Harcourt, River State, Nigeria;
fasten spoilage of the vegetables. High numbers of E-Mail: mac2finney@yahoo.com;
these microorganisms in raw consumed and salad iheanyi.okonko@uniport.edu.ng;
produce would lead to the consumer’s illness with Tel.: +234-080-3538-0891
attendant symptoms and consequences of the
particular or combined microbial presence. Reduction REFERENCES
of risk for human illness associated with raw product 1. Adeleke OE and Odela HA. Plasmid profiles of
can be better achieved through controlling points of multiple drug resistant local strain of
potential contamination in the field during harvesting, Staphylococcus aureus .American Journal of
during processing or shipment, storage or distribution Medical Science. 1997; 20 pp 111-121.
in the retail markets food services facilities or home 2. Akinmusire OO. Fungal Species Associated with
(Beuchat and Ryn, 1997) the Spoilage of Some Edible Fruits in Maiduguri
The presence of indicator and other Northern Eastern Nigeria. Advances in
organisms examined in this study is of special concern Environmental Biology, 2011; 5(1): 157-161.
and perhaps the greatest danger associated with food 3. Akintobi AO, Okonko IO, Akano OR, Agubiade
for human consumption is contamination by human SO, Onianwa O. Isolation and identification of
excrement (Okonko et al., 2008a,b,c, 2009a,b). The fungi associated with the spoilage of some selected
need for microbial assessment of vegetables for fruits in Ibadan, South Western Nigeria. Academia
production of food salads and for other use should also Arena, 2011; 3(11): 1-10
be emphasized to reduce possible contamination 4. Al-Hindi RR, Al-Najada AR, Mohamed SA.
(Fagade et al., 2005). The isolation of these fungi Isolation and identification of some fruit spoilage
pathogens confirmed the studies of Gupta and Pathak fungi: Screening of plant cell wall degrading
(1986), Baiyewu (1994), Baiyewu and Amusa (1999), enzymes. African Journal of Microbiology
Effiuvwevwere (2000), Chulze (2006), Oelofse Research, 2011; 5(4): 443-448.
(2006), Yildz and Baysal (2006), Baiyewu et al. 5. Asagbra A and Oyewole OB. Fermentation studies
(2007), Chuku et al. (2008), Bali et al. (2008), on carrot juice processed to table wine. Nigeria‘s
Okereke et al. (2010), Chukwuka et al. (2010), Al- food Journal, 2002; 20:74-77.
Hindi et al. (2011), Akinmusire (2011) and Akintobi et 6. Badosa E, Trias R, Parés D, Pla M, Montesinos E.
al. (2011) that Rhizopus spp., and A. niger found Microbiological quality of fresh fruit and
associated with rotten Pawpaw are highly pathogenic vegetable products in Catalonia (Spain) using
causing appreciable losses in Pawpaw fruits at post normalised plate-counting methods and real time
harvest. polymerase chain reaction (QPCR). J Sci Food
Although salad vegetables are commonly Agr 2008; 88:605-611
associated with food poisoning, they harbour disease 7. Baiyewu RA, Amusa NA, Ayoola OA, Babalola
causing organisms. The growth of these in the OO. Survey of the post harvest diseases and
environment such oil which is a rich source of aflatoxin contamination of marketed Pawpaw fruit
microbes. Poor agricultural practices such as irrigation (Carica papaya L) in South Western Nigeria.
with contaminated water also may introduce African Journal of Agricultural Research, 2007;
microorganisms. Poor storage and transportation 2(4): 178-181.

29
 Nature and Science, 2012;10(3) http://www.sciencepub.net/nature

8. Baiyewu RA, Amusa NA. Biochemical changes in 19. Davis S. Detection of Methicilin – Resistant
Pawpaw fruits (VAR. ISOLO, JS22 and Staphylococcus aureus, the evaluation of rapid
HOMESTEAD) infected with fungi Bioscience agglutination method. B. J. Biomed science, 1997;
Research Communications, 1999; 11(3): 257-261. 54;13-15.
9. Baiyewu RU. Fungi associated with fruit rot of 20. Dharmaputra, OS, ASR Putri. The relation
Pawpaw (Carica papaya L.) in Southwestern between splitted kernels and population of storage
Nigeria. PhD. Thesis, University of Ibadan, fungi in peanuts. SEAMEO BIOTROP.
Nigeria, 1994; pp.145. Unpublished, 1997.
10. Bali RV, Bindu MG, Chenga RV, Reddy K. Post 21. Domsch KH, Gams W, Anderson TH. Compedium
harvest fungal spoilage in sweet orange (Citrus of Soil Fungi. Academic Press., London, 1993; p.
sinensis) and acid lime (Citrus aurentifolia 860.
Swingla) at different stages of marketing. 22. Droby S. Improving quality and safety of fresh
Agricultural Science Digest., 2008; 28: 265-267. fruits and vegetables after harvest by the use of
11. Bankole MO, Oladimeji DS, Omemu AM. biocontrol agents and natural materials. Acta
Microorganisms associated with the palms of Horticulture, 2006; 709: 45–51.
food-vendors in Abeokuta Metropolis. In: the 23. Dunn RA, Hall WN, Altamirano JV, Dietrich SE,
Book of Abstract of the 29th Annual Conference & Robinson-Dunn B, Johnson DR. Outbreak of
General Meeting (Abeokuta 2005) on Microbes As Shiegella flexneri linked to salad prepared at a
Agents of Sustainable Development, organized by central commissary in Michigan. Public Health
Nigerian Society for Microbiology (NSM), Reports, 1995; 110 (5): 580-586
University of Agriculture, Abeokuta, from 6-10 th 24. Eckert JW, Ogawa JM. The chemical control of
November, 2005. p17 postharvest diseases: deciduous fruits, berries,
12. Barth M, Hankinson TR, Zhuang H, Breidt F. vegetables and root/tuber crops. Annual Review
Microbiological Spoilage of Fruits and Vegetables. Phytopathology, 1988; 26, 433–469.
W.H. Sperber, M.P. Doyle (eds.), Compendium of 25. Effiuvwevwere BJO. Microbial Spoilage Agents
the Microbiological Spoilage of Foods and of Tropical and Assorted fruits and Vegetables
Beverages, Food Microbiology and Food Safety. C (An Illustrated References Book). Paragraphics
Springer Science+Business Media, LLC, 2009; publishing company, Port Harcourt. 2000;pp: 1-
pp135-183. 39.
13. Bulaong SSP, Dharmaputra OS. 2002. Fungal 26. Fagade OE, Ogunjobi AA, Oyelade AA (2005).
Population, Aflatoxin And Free Fatty Acid Microflora of non-carbonated orange drink. In:
Contents Of Peanuts Packed In Different Bag the Book of Abstract of the 29th Annual
Types. Biotropia NO. 19 : 1 – 25 Conference & General Meeting (Abeokuta 2005)
14. Burnett and Beuchat. Emerging infectious on Microbes As Agents of Sustainable
Diseases. Produce Handling and Processing Development, organized by Nigerian Society for
Practices. 2001; 5:6 Microbiology (NSM), UNAAB, from 6-10th Nov.,
15. Carlin F, Nguyen CO, Cudenned P, Reich M. 2005. p16
Microbiolgical Spoilage of fresh, ready to use 27. Food and Safety Authority of Ireland Scientific
grated carrots. Sci. Alim., 1989; 9: 391-396. Criteria to ensure safe food. Institute of Medicine
16. Chuku EC, Ogbonna DN, Onuegbu BA, Adeleke National Research Council. The National
MTV. Comparative Studies on the Fungi and Bio- Academies Press, Washington, 2001.
Chemical Characteristics of Snake Gaurd 28. Frisvad JC, Samson RA. Mycotoxins produced by
(Trichosanthes curcumerina linn) and Tomato species of Penicillium and Aspergillus occurring in
(Lycopersicon esculentus mill) in Rivers state, cereals.. In: Chelkowski., J.(ed.) Cereal Grain.
Nigeria. Journal of Applied Sciences, 2008; 8(1): Mycotoxins, Fungi and Quality in Drying and
168-172. Storage. Elsevier, Amsterdam fungi. Burgess
17. Chukwuka, KS, Okonko, IO, Adekunle AA. Publishing Company. USA, 1991; pp. 441-476.
Microbial Ecology Of Organisms Causing 29. Goldblatt LD and Stoloff L. History and natural
Pawpaw (Carica Papaya L.) Fruit Decay in occurrence of aflatoxins. Proceedings of the
Oyo State, Nigeria. American-Eurasian Journal of International Symposium on Mycotoxins, (Naguib
Toxicological Sciences, 2010; 2 (1): 43-50 K, Naguib MM, Park DL and Pohland AE. Eds.).
18. Chulze SN, Magnoli CE, Dalcero AM. Occurrence The Gen. Organ. for Govern. Printing Offices.
of ochratoxin A in wine and ochratoxigenic Cairo, Egypt, 1983; pp. 3346.
mycoflora in grapes and dried vine fruits in South 30. Gupta AK, Pathak VN. Survey of fruit market for
America. Intl. J. Food Microbiol., 2006;111:S5-S9. papaya fruit rot by fungi pathogens. Indian J.
Mycol., 1986; 16:152-154.

30
 Nature and Science, 2012;10(3) http://www.sciencepub.net/nature

31. Ho JL, Shands KN, Freidland G, Eckind P, Fraser, 44. Nguyen CF. The Microbiology of Minimally
DW. An outbreak of type 46 Listeria Processed Fresh Fruits and Vegetables. Critical
monocytogenes Infection involving Patients from Review in Food Science and Nutrition, 1994;
Eight Boston Hospital. Archives of internal 34:371-401.
Medicine, 1986; 146:520-524, 45. Oelofse D, Dubery IAM, Arendse S, Mm S,
32. Hung HC, Joshipura KJ, Jiang RH, Hunter D, Gazendam I, Berger DK. Apple polygalacturonase
Smith SA. Fruits and Vegetable intake and the risk inhibiting protein expressed in transgenic tobacco
of major chronic disease. Journal of National inhibits polygalacturonases from fungal pathogens
Cancer Institute, 2004; 95:157- 164. of apple and the anthracnose pathogen of lupins,
33. ICMSF (International Commission on Phytochemistry, 2006; 67: 255-263.
Microbiological Specification for Foods). 46. Okereke VC, Godwin-Egein MI, Arinze AE.
Microorganisms in foods, vol6 Microbial Ecology Assessment of Postharvest Rot of Mango at
of food commodities, New York; Blackie Different Stages of Market in Port Harcourt,
Academics and Professional, 1998 617 Nigeria. Int. J. Curr. Res., 2010; 11: 6-10.
34. Jay, M.J. (2005). Modern Food Microbiology 4 th 47. Okonko IO, Ogun AA, Adejoye OD, Ogunjobi
Edtn. Chapman and Hall, New York.p.187. AA, Nkang AO, Adebayo-Tayo BC. Hazards
35. Jiwa SFH, Kiovacek K, Wadstorm T. analysis critical control points (HACCP) and
Enterotoxigenic Bacteria in Food and Water from Microbiology qualities of Sea-foods as affected by
an Ethiopian Community .Applied Environmental Handler’s Hygiene in Ibadan and Lagos, Nigeria.
Microbiology, 1981; 41:105. African Journal of Food Sciences, 2009a;
36. Jolt JG, Krieg NR, Sneath PHA, Stanley JT, 3(1):035-050
Williams ST. 1994. Bergey’s manual of 48. Okonko IO, Donbraye E, Babatunde SOI.
systematic bacteriology, 9th edn. Williams & Microbiological Quality Seafood Processors and
Wilkins Co. Baltimore, Maryland, p786 Water Used in Two Different Sea Processing
37. Kaneko K, Hayashidani H, Ohromo Y, Kousge J, Plants in Nigeria. EJEAFChe, 2009b; 8(8):621-
Kato M, Takahashi K, Shiraki Y, Ogawa M. 629
Bacterial Contamination of ready to eat foods and 49. Okonko IO, Ogunnusi TA, Adejoye OD, Shittu
fresh products in retail shops and food factories. OB. Microbiological and Physicochemical
Journal of food protection, 2003; 62(6): 644-649. Analysis of Different Water Samples Use for
38. Klich MA. Identification of common Aspergillus Domestic Purposes in Abeokuta, Ogun State and
species. CBS, Utrecht., 2002; p. 116. Ojota, Lagos State, Nigeria. African Journal of
39. Kozakiewicz Z. Occurrence and significance of Biotechnology, 2008a; 7 (5):617-621
storage fungi and associated mycotoxins in rice 50. Okonko IO, Ogunjobi AA, Fajobi EA, Onoja BA,
and cereal grains. In: Highley E, GI Johnson Babalola ET, Adedeji AO. Comparative studies
(eds.). ACIAR Technical Reports 37. Mycotoxin and Microbial risk assessment of different Ready-
Contamination in Grains. Paper presented at the to-Eat (RTE) frozen sea-foods processed in Ijora-
17th ASEAN Technical Seminar on Grain olopa, Lagos State, Nigeria. African Journal of
Postharvest Technology. Lumut, Malaysia, 25-27 Biotechnology, 2008b; 7(16): 2898-2901
July 1995; 1996; pp. 18-26. 51. Okonko IO, Ogunjobi AA, Adejoye OD, Ogunnusi
40. Krogh P. Adverse effect of mycotoxins on human TA, Olasogba MC. Comparative studies and
health in: seed pathology. In Mathur, S. B. and Microbial risk assessment of different water
Jorgensen, J. (Eds), Proceedings of the seminar, 20- samples used for processing frozen sea-foods in
25 June 1988, Copenhagen, Denmark, 1992; pp. Ijora-olopa,Lagos State, Nigeria. African Journal
149-57. of Biotechnology, 2008c; 7(16):2902-2907
41. Lelieveld HLM, MA Mostert, J Holah and B 52. Okonko IO, Ogunnusi TA, Ogunjobi AA, Adedeji
White. Hygiene in food processing. Woodhead AO, Adejoye OD, Babalola ET, Ogun AA.
Publishing in Food Science and Technology. Microbial studies on frozen shrimps processed in
England, 2003. Ibadan and Lagos, Nigeria. Scientific Research
42. Mgbakor C, Ojiegbe GC, Okonko IO, Odu NN, Alli and Essay, 2008d; 3(11): 537-546
JA, Nwanze JC Onoh CC. Bacteriological 53. Omemu AM, Bankole MO. Ready-to-eat (RTE)
evaluation of some sachet water on sales in Owerri vegetable salad: effect of washing and storage
metropolis, Imo State, Nigeria. Malaysia Journal of temperature on the microbial quality and shelf-life.
Microbiology, 2011; 7(4): 217-225 In: the Book of Abstract of the 29th Annual
43. Monso EM. Occupational asthma in greenhouse Conference & General Meeting (Abeokuta 2005)
workers, Curr. Opin. Pulm. Med., 2004; 10: 147-150. on Microbes As Agents of Sustainable
Development, organized by Nigerian Society for

31
 Nature and Science, 2012;10(3) http://www.sciencepub.net/nature

Microbiology (NSM), UNAAB, from 6-10th Nov, 67. Sauer DB, RA Meronuck, CM Christensen.
2005. p28 Microflora. In: Sauer, D.B. (ed.). Storage of Cereal
54. Ongeng D, Devlieghere F, Debevere J, Coosemans Grains and Their Products. Amer. Assoc. Cereal
J, Ryckeboer J. The efficacy of electrolysed Chemists, Inc. St. Paul, Minnesota, 1992; pp. 313-
oxidizing water for inactivating spoilage 340.
microorganisms in process water and on minimally 68. Stinson EE, Osman SF, Heisler EG, Siciliano J,
processed vegetables. Int J Food Microbiol, 2006; Bills DD. Mycotoxin production in whole
109:187-197 tomatoes, apples, oranges and lemons. J. Agric.
55. Onyia VN, Mbuka CO, Ihejirika GO, Obilor OP, Food Chem., 1981; 29: 790-792.
Duruigbo CI, Onweremadu EC. Studies on the 69. Tournas VH, Stack ME. Production of alternariol
Performance and Incidence of Fusarium wilt of and alternariol methyl ether by Alternaria
Tomatoes under different colours of plastic mulch. alternata grown on fruits at various temperatures.
Nig. Soc. Plant proc. 32nd Ann. Conf. Book of J. Food Prot., 2001; 64: 528-532.
Abstract, 2005; pp: 23-25. 70. Trias R, L Bañeras, E Montesinos, E Badosa.
56. Oyeleke SB, Manga SB. Essentials of Laboratory Lactic acid bacteria from fresh fruit and vegetables
Practicals in Microbiology. Tobest publisher, as biocontrol agents of phytopathogenic bacteria
Minna, Nigeria, 2008; pp.36-75. and fungi. International Microbiology, 2008;
57. Peraica M, Radic B, Lucic A, Pavlovic M. Toxic 11:231-236
effects of mycotoxins in humans. Bull. World 71. Yildz H, Baysal T. Effects of alternative current
Health Organ., 1999; 77: 754-766. heating treatment on Aspergillus niger, pectin
58. Peraica M, Radic B, Lucic A, Pavlovic M. Toxic methylesterase and pectin content in tomato. J.
effects of mycotoxins in humans. Bull. World Food Eng., 2006; 75: 327-332.
Health Organization, 1999; 77: 754-766. 72. Yusuf IZ, Oyaweye OM, Yongabi KA, Pemu AT.
59. Petzinger E, Weidenbach A. Mycotoxins in the Bacteriological Quality Assessment of Salad
food chain: The role of ochratoxins. Livest. Prod. Vegetables sold in Bauchi Metropolis. Nigeria
Sci., 2002; 76: 245-250. Journal of Microbiology, 2004;18:316-320.
60. Pitt JI, Hocking AD. Fungi and food spoilage. 73. Zhu SJ. Non-chemical approaches to decay control
Blackie Academic and Professional, London, UK, in postharvest fruit. In: Noureddine, B., Norio, S.
1997. (Eds.), Advances in Postharvest Technologies for
61. Pitt JI, AD Hocking. Current knowledge of fungi Horticultural Crops. Research Signpost,
and mycotoxin assessment of food commodities in Trivandrum, India, 2006; pp. 297–313.
South East Asia. In: Highley, E. and G.I. Johnson
(eds.). ACIAR Technical Reports 37. Mycotoxin
Contamination in Grains. Paper presented at the
17"' ASEAN Technical Seminar on Grain
Postharvest Technology. Lumut, Malaysia, 1995
July 25-27, 1996; pp. 5-10
62. Pitt JI, AD Hocking, BF Miscamble, OS
Dharmaputra, KR Kuswanto, ES Rahayu and
Sardjono. The mycoflora of food commodities
from Indonesia. Journal of Food Mycology, 1998;
1(1): 41-60.
63. Pomeranz Y. Biochemical, functional and
nutritional changes during storage. In: Sauer DB
(ed.). Storage of Cereal Grains and Their Products.
Amer. Assoc. Cereal Chemists, Inc. St. Paul,
Minnesota, 1992; pp. 55-141.
64. Prasad T. Plant pathogenesis and disease control.
Plant Dis. J. of Japan Acado., 1992; 56:367.
65. Rosenblum LS, Mirkin IR, Allen FT, Safford S,
Haller SC. A trace to community distributed
lettuce. American Journal of Public Health, 1990;
80; 1075-1079.
66. Samson RA, Varga J. Aspergillus systematics in
the genomic era. CBS Fungal Biodiversity Centre,
Utrecht, 2007; p. 206.

32
 Nature and Science, 2012;10(3) http://www.sciencepub.net/nature

2/2/2012

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