MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS OF

SEAFOOD IN GREECE

C. PAPADOPOULOU1,5, E. ECONOMOU1, G. ZAKAS1,2, C. SALAMOURA1, C.

DONTOROU1,3 and J. APOSTOLOU1,4
1
Food Microbiology Unit, Microbiology Department Medical School, University of
Ioannina Dourouti University Campus 45110 Ioannina, Greece

2
Preveza Aquaculture Station
Ministry of Agriculture
Preveza, Greece

3
State Veterinary Service
Prefecture of Ioannina, Greece

4
Regional Veterinary Laboratory Service
Ministry of Agriculture
Ioannina, Greece

Accepted for Publication November 11, 2006

ABSTRACT

A total of 360 samples, including 105 marine fish, 25 prawns, 50 squid, 50

octopus, 30 mussels and 100 freshwater fish were examined microbiologi-cally
for the presence of microorganisms potentially pathogenic. All samples were
examined following standard microbiological procedures. The isolated
microorganisms were: Aeromonas hydrophilia (38–93%), Klebsiella ozonae (1–
40%), Escherichia coli (14–87%), Yersinia enterocolitica (0–40%), Hafnia alvei
(0–36.6%), Enterobacter agglomerans (0–42%), Citrobacter freundii (0–46%),
Proteus vulgaris (15–80%), Proteus mirabilis (7–82%), Morganella morganii (0–
30%), Pseudomonas fluorescens (0–34%), Pseudomonas putida (0–6%),
Plesiomonas shigelloides (0–4%), Listeria innocua (1–3.3%), Vibrio
parahaemolyticus (0–2%), Clostridium perfringens (0–1%), Staphylococcus
aureus (0–80%) and Candida quillermondi (0–1%), Candida albicans (0–1%),
Penicillium oxalicum (0–1%) and Penicillium italicum (0–12%).

5
Corresponding author. TEL: +3026510-97592; FAX: +3026510-93563; EMAIL: cpapadop@
cc.uoi.gr

Journal of Food Quality 30 (2007) 28–42. All Rights Reserved.

28 © 2007, The Author(s)
Journal compilation © 2007, Blackwell Publishing
 MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS 29

PRACTICAL APPLICATIONS

Marine and fresh water fish used not to be considered important vectors
of human pathogens. However, this situation is changing, due to
environmental pollution and increasing animal densities as a consequence of
a rapidly growing food industry and also due to increasing awareness by
health care professionals on seafood pathogens that may cause human illness.
The pres-ence of pathogenic microorganisms represents a safety concern,
particularly if seafood is eaten raw. In the present study, the presence of a
wide range of pathogens, most of which are associated with foodborne
diseases, indicates the potential health risks from contaminated seafood.
Although most of the iso-lated genera are heat-labile and adequate cooking
will inactivate them, improper handling and cross-contamination or raw
seafood eating habits, might pose a health hazard, especially to susceptible
population such as the immunosupressed, children and the elderly.

INTRODUCTION

The recent food scares related to ruminant’s meat (bovine spongiform

encephalitis [BSE], scrapie), chicken meat (Salmonellae, avian influenza,
dioxins) and the dietary changes of the consumers toward healthier food habits
(e.g., Mediterranean diet, sushi), has resulted in a rapid rise of seafood con-
sumption worldwide (Wallace et al. 1999; Yasumoto 2000). However, as
demands for and production of seafood increased, it became important to pay
more attention to seafood safety and seafood-related diseases. Although they
were problems previously regarded as local and insignificant, now it is well
known that they can spread to other regions as a result of increasing world trade
and environmental changes. Furthermore fish and shellfish are eaten raw in
many cultures and this habit provides the opportunity for direct transmission of
pathogens. Contaminated seafood has been implicated in foodborne out-breaks
in many countries (Jensen and Greenlees 1997; Lipp and Rose 1997; Wallace et
al. 1999; Rocourt et al. 2000; Gillespie et al. 2001), the majority of them having
been caused by bacteria, viruses, histamine and marine biotoxins.
Although there is some information on the incidence of various patho-
gens on fish (Fricker and Tompsett 1989; Gobat and Jemmi 1993; Walker
and Brooks 1993; Hanninen et al. 1997; Davies et al. 2001) the data
provided are either restricted to a few pathogens, or to a limited geographical
area. The objective of this survey was to generate information on the
prevalence of pathogenic and potentially pathogenic microorganisms, which
under suitable conditions may cause mild to severe public health problems.
30 C. PAPADOPOULOU ET AL.

MATERIALS AND METHODS

Sampling
Within a period of 12 months (from August 2004 to August 2005) 360
randomly collected samples were analyzed microbiologically.
There was a total number of 260 seafood samples examined counting 25
from prawns (Penaeus kerathurus), 50 from squids (Loligo vulgaris), 50 from
octopuses (Octapus vulgaris), 30 from mussels (Mytilus galloprovincialis) and
105 from fresh marine fish including: sardines (Sardina pilchardus), plaices
(Solea vulgaris), hakes (Merluccius merluccius), bogues (Boops boops), silver
sides (Atherina hespectus), mullets (Mugil cephalus) and mackerels (Scomber
scomber, Scomber colias). Also a total number of 100 freshwater fish were
examined. The freshwater fish samples were trout (Salmo truta), collected from
local fish retail stores. All samples were obtained from retail stores of
northwestern Greece and were of local capture (from the Ionian Sea). The
samples were collected within 24 h of capture, placed into sterile containers and
were transferred to the laboratory at 4C, using insulated transport boxes. The
samples were either processed upon delivery to the laboratory or were packed in
sterile ice and stored inside a cold ward at 4C for 24 h prior to the
microbiological analysis.

Microbiological examination
All samples were analyzed for the following microbial pathogens:
Vibrio spp., Salmonella spp., Listeria spp., enteric indicator organisms
(members of the Enterobacteriacae family and Enterococci), Pseudomonas
spp., Clostridium perfringens, Staphylococcus spp., yeasts and moulds.
The microbiological procedures included the homogenization of 25 g of
each sample (flesh from seafoods or fish flesh and skin), in 225 mL of appro-
priate broth according to standard microbiological methods (ISO methods). The
methods specifically used were: for Salmonella spp. the ISO 6579-6579 (E); for
Staphylococcus spp. the ISO 6888-1983 (E); for C. perfringens the ISO 7937-
1997 (E); for Enterobacteriacae, culture on violet red bile glucose agar
(VRBGA) according to ISO 7402-1993 (E); for Yersinia enterocolitica the ISO
10273-1994 (E); for Enterococcus spp., culture according to method 1 as
described by Roberts et al. (1995); for Pseudomonas spp., culture accord-ing to
method 3 as described by Roberts et al. (1995); for Vibrio spp., enrich-ment
culture according to method 1 as described by Roberts et al. (1995); for Listeria
spp., pre-enrichment in half-Fraser broth and enrichment in Fraser broth
(FRASER Listeria Selective Enrichment Broth, Merck, Darmstadt, Germany)
followed by cultures on to Oxford agar (OXFORD Listeria Selec-tive Agar Base,
Merck) and PALCAM agar (PALCAM Listeria Selective Agar
 MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS 31

Base, Merck) as recommended by ISO-10560-1999 (E); and for yeasts and

moulds, the ISO 7954-1987 (E) was employed. Identification of the bacteria
and yeast isolates to the species level was performed using the API-20E,
API-NE, API-STAPH, API LISTERIA and API-CHAUX (bioMerieux,
Marcy l’ Etoile, France). For moulds, species identification was based on the
micro-scopic appearance and colony morphology.

RESULTS

The results are presented in Table 1. Concerning pathogens causing

severe infections, Salmonella spp. was not detected in any of the samples
examined, V. parahaemolyticus was detected in two samples of fish (bogue
and cod), Listeria innocua was detected from two samples (from mussels and
river trout) and C. perfringens was detected in one sample of freshwater fish
(trout). Aeromonas hydrophila was the most commonly isolated species,
followed by P. vulgaris, P. mirabilis, Morganella morganii and other
members of the Enterobacteriacae family. Plesiomonas shigelloides was
isolated from both marine and freshwater fish. Yeasts and moulds were
isolated only from fresh-water fish samples, with the exception of one strain
of Candida isolated from marine fish. Mussels were the most contaminated
seafood, showing the highest frequencies of Escherichia coli (100%), P.
vulgaris (96%) and P. mirabilis (92%) isolations and harboring pathogens of
public health importance such as Y. enterocolitica (40%), Staphylococcus
aureus (56.6%), Pseudomonas fluorescens (26.6%) and L. innocua (3.3%).

DISCUSSION

A. hydrophila, Pseudomonas spp., V. parahaemolyticus, most members

of the enterobacteriacae family, Listeria spp., Yersinia spp. and S. aureus, are
indigenous in aquatic environments and are found in large numbers particu-
larly in polluted aquatic environments. Such species are expected to be
common fish contaminants and consist a potential food safety hazard; there-
fore our results show that this is the case for most of the bacterial species
isolated. However, the yeasts (Candida spp.) and moulds (Penicillium spp.)
species isolated, especially the latter, are rather unusual to occur in the
aquatic environment, and any foodborne transmission is yet to be proven or
questioned.
The high incidence of A. hydrophila isolations from almost all marine
(93%) and freshwater fish (38%) and other seafood (78–86%) examined in this
32 C. PAPADOPOULOU ET AL.

TABLE 1.
INCIDENCE OF MICROORGANISMS ISOLATED IN SEAFOOD AND SEAWATER
IN GREECE

Microorganism Incidence %
(Number of positive samples/number of tested samples)

Bacteria Seafood Marine fish Freshwater fish

Aeromonas hydrophila 80.0 (20/25) prawns 93.0 (98/105) 38.0 (38/100)

86.0 (43/50) squids
78.0 (39/50) octopus
73.3 (22/30) mussels
Proteus vulgaris 60.0 (15/25) prawns 45.0 (47/105) 15.0 (15/100)
22.0 (11/50) squids
26.0 (13/50) octopuses
80.0 (24/30) mussels
Proteus mirabilis 32.0 (8/25) prawns 51.0 (54/105) 7.0 (7/100)
26.0 (13/50) squids
14.0 (7/50) octopuses
76.6 (23/30) mussels
Pseudomonas fluorescens 0.0 (0/25) prawns 34.0 (36/105) 15.0 (15/100)
8.0 (4/50) squids
10.0 (5/50) octopuses
26.6 (8/30) mussels
Pseudomonas putida 0.0 (0/25) prawns 6.0 (6/105) 0.0 (0/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Plesiomonas shigelloides. 0.0 (0/25) prawns 4.0 (4/105) 2.0 (2/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Enterobacter agglomerans 8.0 (2/25) prawns 42.0 (44/105) 0.0 (0/100)
2.0 (1/50) squids
0.0 (0/50) octopuses
36.6 (11/30) mussels
Escherichia coli 32.0 (8/25) prawns 87 (91/105) 14.0 (14/100)
6.0 (3/50) squids
18.0 (9/50) octopuses
83.3 (25/30) mussels
Citrobacter freundii 0.0 (0/25) prawns 46.0 (47/105) 1.0 (1/100)
2.0 (1/50) squids
4.0 (2/50) octopuses
33.3 (10/30) mussels
Morganella morganii 0.0 (0/25) prawns 30.0 (31/105) 1.0 (1/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
20.0 (6/30) mussels
Klebsiella ozonae 0.0 (0/25) prawns 40.0 (42/105) 2.0 (2/100)
6.0 (3/50) squids
4.0 (2/50) octopuses
40.0 (12/30) mussels
 MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS 33

TABLE 1. CONTINUED
Microorganism Incidence %
(Number of positive samples/number of tested samples)

Bacteria Seafood Marine fish Freshwater fish

Hafnia alvei 24.0 (6/25) prawns 20.0 (21/105) 1.0 (1/100)

0.0 (0/50) squids
4.0 (2/50) octopuses
36.6 (11/30) mussels
Yersinia enterocolitica 0.0 (0/25) prawns 10.0 (10/105) 0.0 (0/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
40.0 (12/30) mussels
Salmonella spp. 0.0 (0/25) prawns 0.0 (0/105) 0.0 (0/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Staphylococcus aureus 8.0 (2/25) prawns 80.0 (84/105) 6.0 (6/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
56.6 (17/30) mussels
Vibrio parahaemolyticus 0.0 (0/25) prawns 2.0 (2/105) 0.0 (0/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Clostridium perfringens 0.0 (0/25) prawns 0.0 (0/105) 1.0 (1/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Listeria spp. 0.0 (0/25) prawns 0.0 (0/105) 1.0 (1/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
3.3 (1/30) mussels
Yeasts and moulds
Candida quillermondi 0.0 (0/25) prawns 0.0 (0/105) 1.0 (1/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Candida albicans 0.0 (0/25) prawns 1.0 (1/105) 0.0 (0/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Penicillium oxalicum 0.0 (0/25) prawns 0.0 (0/105) 1.0 (1/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Penicillium italicum 0.0 (0/25) prawns 0.0 (0/105) 1.0 (1/100)
0.0 (0/50) squids
0.0 (0/50) octopuses
0.0 (0/30) mussels
Samples examined = 360 155 105 100
34 C. PAPADOPOULOU ET AL.

study, concurs with the results of other researchers in the U.K., New
Zealand, Switzerland, Taiwan, Portugal, France and Greece (Fricker and
Tompsett 1989; Hudson et al. 1992; Gobat and Jemmi 1993; Tsai and Chen
1996; Hanninen et al. 1997; Davies et al. 2001), who have found incidences
ranging from 19% to 90% in similar samples. Aeromonas species have been
recog-nized as potential or emerging foodborne pathogens for more than 20
years, and A. hydrophila has been occasionally associated with foodborne
disease, the clinical manifestations being either extraintestinal (sepsis,
meningitis, peri-tonitis, endocarditis, pneumonia, ocular and urinary tract
infections, septic arthritis, osteomyelitis and soft-tissue infections) or
gastroenteritis (Hudson et al. 1992; Kirov 1997; Isonhood and Drake 2002).
Nevertheless, A. hydro-phila, like most aeromonads, is psychrotrophic, able
to grow in foods during cold storage, it is not resistant to food processing
regimes and is readily killed by heat treatment, thus any risk of foodborne
infection results from eating raw or undercooked seafood.
Ps. fluorescence (0–34%), Pseudomonas putida (0–6%) and
Plesiomonas spp. (0–4%) were isolated from a small number of samples. Ps.
fluorescence and Ps. putida are common contaminants of both marine and
freshwater fish, accounting for fish spoilage (Van Damme and Vandepitte
1980; Vandepitte et al. 1980; Gennari and Dragotto 1992; Gram 1993;
Smolowitz et al. 1998; Tryfinopoulou et al. 2002). Plesiomonas shigelloides
is a common pathogen in tropical regions, but it is rarely isolated in
temperate climates; it seems to be ubiquitous in freshwater worldwide and
may also cause invasive infections in cold-climate areas. It is often isolated
from surface water and fish, and has been found to cause gastroenteritis. The
course of infection is sometimes asymptomatic, but usually, patients develop
an acute gastroenteritis, while immunocompromised patients can show
serious courses of infection. Bacter-emia caused by P. shigelloides, although
a rare event, is often associated with the consumption of seafood and fresh or
estuarine water in temperate or tropical climates. However most patients with
bacteraemia have shown under-lying health disorders (Jonsson et al. 1997;
Aldova et al. 1999; Aldova 2000; Knebel et al. 2001).
Various members of the Enterobacteriacae family can be found in the
aquatic environment, some of them are recognized as indicator microorgan-
isms of fecal pollution. E. coli and Proteus spp. are commonly isolated from
seafood. Therefore, Proteus species, E. coli and some other enteric bacteria
like Morganella, Klebsiella, Hafnia, Enterobacter, Serratia and Citrobacter
are important histamine-producing bacteria in fish with high free histidine
content, especially in scombroid fish, and have been implicated in histamine
food poisoning (or scombroid poisoning) incidents (Taylor 1986; Ababouch
et al. 1991; Rodriguez-Jerez et al. 1994). However, in the present survey
histamine production was not studied.
 MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS 35

Y. enterocolitica is an enteric pathogen associated with a wide range of

clinical and immunological signs and symptoms principally reported in chil-
dren, but can occur in any age group. It is widely distributed in the environ-
ment, and is more frequently isolated in temperate areas because of its
psychrotrophic nature enabling it to survive in the aquatic environment in
temperatures as low as 0C. However, in the recent 12 years the organism has
been recognized as an emerging foodborne pathogen, next to Salmonella and
Campylobacter (Lanser et al. 1993; Fenwick and McCarthy 1995), contami-
nating a variety of food including seafood (Barton et al. 1997). Although the
incidence of Y. enterocolitica isolation from fish has been found to be within
the range of 0–23% (Hudson et al. 1992; Walker and Brooks 1993; Khare et
al. 1996; Velazquez et al. 1996; Davies et al. 2001) and in our survey, from
0–10% (freshwater fish and sea fish, reciprocally) to 40% (mussels), fish
consumption has not been the cause of any proven case of human yersiniosis
yet.
Listeria spp. is a gram-positive, aerobic or microaerophilic rod, that has
been isolated from humans, a variety of animals, environmental (soil, water)
samples, forages, fresh and ready-to-eat foods. The listeria species most fre-
quently isolated from seafood are L. innocua, Listeria monocytogenes and
Listeria welshimeri, and the reported incidence ranges from 0–56%. L.
innocua is the most common species encountered in fresh and cold-smoked
salmon, mussels, squids, mackerels and other seafood (Farber 1991; Dillon
et al. 1992, 1994; Rorvik et al. 1995; Vaz-Velho et al. 1998, 2000; Laciar
and Centorbi 2002). In our study, two strains (0.6%) of L. innocua were
isolated from mussels (3.3%) and trouts (1.0%). Although L. innocua is
referred as a nonpathogen for humans in microbiology textbooks, recently it
has been described in association with human disease. A case of fatal bac-
teremia caused by L. innocua in a 62-year-old patient was reported in 2003 in
France (Perrin et al. 2003). This is not the only case of a nonpathogen
turning into a pathogen; in clinical microbiology all microbes are considered
as “pathogens or potential pathogens” for humans. The phenomenon of non-
pathogenic microorganisms turning into pathogens is attributed to the
increased ability of the microorganisms to adapt to new hosts and to the
increased numbers of immunosupressed or immunodeficient subjects who
are vulnerable to opportunistic infections. Isolates characterized as “non-
pathogenic for humans” should be very carefully estimated in reports to
regulatory agencies, especially those species that are already known to be
pathogenic for animals.

Salmonella was not detected in any sample during this study. In similar
studies carried out by other researchers elsewhere, Salmonella has also not
been isolated (D’Aoust et al. 1980; Belchior and Pucci 2000; Davies et al.
2001) or has been occasionally isolated (Youssef et al. 1992). Salmonella is
36 C. PAPADOPOULOU ET AL.

not indigenous to the aquatic environment, it is not a psychrophilic organism

and normally it is found in the intestinal tract of a wide variety of animals
and in humans. So Salmonella can be shed through feces to the environment
and thus could contaminate soil, pasture, streams, lakes, rivers and seas.
Seafood can be contaminated from salmonella when captured in polluted and
inshore seawaters, or during handling from contaminated personnel. In a 9-
year survey by the US Food and Drug Administration (FDA) on the
incidence of Salmo-nella in 11,312 samples of import and 768 samples of
domestic seafood, the overall incidence was found to be 7.2 and 1.3%,
respectively (Heinitz et al. 2000). Also, food wastes and animal manure may
be fed to farmed fish and crustaceans, and brackish waters may be used for
aquacultures, potentially contaminating such products with salmonella
(Reilly and Twiddy 1992; Jay et al. 1997).
V. parahaemolyticus is causing sporadic foodborne infections and out-
breaks worldwide, with gastroenteritis being the most common clinical mani-
festation. It is indigenous to the marine environment and its survival is
affected by temperature, not surviving in seawater below 10C
(Desmarchelier 1997). Seawater temperatures in Greece are ideal for survival
and growth of Vibrio parahaemolyticus and, in the study carried out by
Davies et al. (2001), it was found in various fish from Greece (anchovy,
bogue, mackerel, mussels, picarel, mullet), in relatively high incidence
(14%), only second to that of Portuguese fish (35%). In the present survey
there were only two isolations of V. parahaemolyticus, (2%) from marine
fish (bogue). Also, in 1995, during the cholera epidemic in the neighboring
countries, 92 seafood samples were routinely examined at the Food
Microbiology Unit, Microbiology Department (Vibrio Reference Laboratory
for Northwestern Greece), and one sample (1%) was found contaminated
with V. parahaemolyticus (Papadopoulou et al. 1996).
S. aureus is part of the normal human and animal microflora and may be
found in an aquatic environment polluted by sewage. Therefore S. aureus is
an important pathogen, causing a wide range of diseases, including food-
borne infections and intoxications. Staphylococcal food poisoning is one of
the commonest types of foodborne disease worldwide usually associated
with meat, poultry and dairy products. The commonest way food is con-
taminated is through time and temperature abuse following improper han-
dling. Because of its halophilic nature, S. aureus can survive in the marine
environment and may contaminate seafood. In the present investigation, the
incidence of S. aureus isolation ranged from 0% (squids and octapuses) to as
high as 56.6% for mussels and 80% for marine fish. However, reports on S.
aureus isolation from fish and seafood are limited. In a study in Argentina on
indicator and foodborne pathogens on hake fillets for export, S. aureus was
not isolated from any sample (Belchior and Pucci 2000), while in another
 MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS 37

study in the U.S., S. aureus was isolated from whole catfish and catfish fillets
(Ramos and Lyon 2000). Nevertheless, the presence of S. aureus in raw fish
is of little public health importance, because it cannot propagate in
competition with fishes’ natural microflora, and, if fish products are involved
in food poisoning, then the origin of the bacteria is the handlers rather than
the fish.
C. perfringens is one of the most widely spread pathogens in the envi-
ronment and an important cause of food poisoning, often associated with
large outbreaks (Pollock and Whitty 1991; Hook et al. 1996). It causes a
variety of symptoms in humans such as gas gangrene, gastroenteritis,
necrotic enteritis and diarrhea in mild cases. C. perfringens is a minor part of
the normal fecal microflora of man and animals, and it is contaminating food
mainly through animal feces, inappropriate handling and storage of prepared
food and tem-perature abuse (Bates 1997). It grows at elevated temperatures
(43–45C) and fresh or frozen seafood is a rather poor vehicle of C.
perfringens food trans-mission. In our study, the incidence of C. perfringens
was very low (1%), and the isolate was detected in a sample of freshwater-
farmed fish (trout). C. perfringens has been isolated from decomposing
skipjack tuna. However, these reports do not concern fresh fish, but canned,
vacuum packed, marinated or abused fish, and this is suggestive of the
potential risks for the public health, owing to the presence of any pathogenic
type of Clostridium spp. in marine or freshwater fish.
Yeasts of the species Candida are opportunistic pathogens, and the iso-
lation of two strains, from marine (1%) and freshwater fish (1%), might be
attributed to mishandling. Fungi are common contaminants of cereals, and
some species are involved in foodborne diseases through the production of
highly toxic mycotoxins. However, the fungal species of Penicillium,
Penicil-lium oxalicum (1%) and Penicillium italicum (1%), isolated from the
fresh-water fish in this study are not toxigenic, and their presence coincides
with contamination as a result of mishandling. Reports on the incidence of
any yeasts or moulds in seafood are very scarce. In a study by Spanish
researchers, the isolation of Candida albicans, Debaryomyces hansenii,
Rodotorula rubra, Rodotorula glutinis and Saccharomyces cerevisiae from
fish is reported (Vazquez-Juarez et al. 1994).
Marine and fresh water fish, either free living or cultured, were not
considered to be important vectors of human pathogens. Then again, this
situation is changing, partly as a result of environmental pollution and
increasing animal densities as a consequence of a rapidly growing food
industry and partly as a result of increasing awareness by health care pro-
viders on seafood pathogens that may result in human illness. Nonetheless,
the presence of pathogenic microorganisms represents a safety concern in
seafood eaten raw and in cases wherein the pathogen has a very low
38 C. PAPADOPOULOU ET AL.

minimum infective dose, such as the case with e.g., Salmonella typhi, Shi-
gella, E. coli O157 Campylobacter and Listeria. For most foodborne infec-
tions and intoxications, it is not simply the presence of certain bacteria
species, but their growth to large numbers that represent a safety risk. In the
present study, the levels of bacterial contamination were not counted for each
species, yet the presence of a wide range of pathogens (A. hydrophila, Y.
enterocolitica, V. parahaemolyticus, S. aureus, C. albicans, C. perfringens),
most of which are associated with foodborne diseases, indicates the potential
risks for the safety of the so-called “healthy” seafood. Although most of the
isolated genera are heatlabile and adequate cooking will inactivate them,
improper handling and cross-contamination or raw seafood eating habits,
might pose a health hazard, especially to susceptible populations such as the
immunosupressed, children and elderly people.

REFERENCES

ABABOUCH, L., AFILAL, M.E., RHAFIRI, S. and BUSTA, F.F. 1991.

Identification of histamine-producing bacteria isolated from sardine
(Sardina pilchardus) stored in ice and at ambient temperature (25C).
Food Microbiol. 8(2), 127–136.
ALDOVA, E. 2000. New serovars of Plesiomonas shigelloides 1992–1998.
Cent. Eur. J. Public Health 8(3), 150–151.
ALDOVA, E., MELTER, O., CHYLE, P., SLOSAREK, M. and KODYM, P.
1999. Plesiomonas shigelloides in water and fish. Cent. Eur. J. Public
Health 7(4), 172–175.
BARTON, M.D., KOLEGA, V. and FENWICK, S.G. 1997. Yersinia entero-
colitica. In Foodborne Microorganisms of Public Health Significance,
5th Ed. (A.D. Hocking, G. Arnold, I. Jenson, K. Newton and P.
Sutherland, eds.) pp 493–519, AIFST NSW Branch, Food Microbiology
Group, North Sydney, Australia.
BATES, J.R. 1997. Clostridium perfringens. In Foodborne Microorganisms
of Public Health Significance, 5th Ed. (A.D. Hocking, G. Arnold, I.
Jenson, K. Newton and P. Sutherland, eds.) pp 407–430, AIFST NSW
Branch, Food Microbiology Group, North Sydney, Australia.
BELCHIOR, E.S. and PUCCI, O.H. 2000. Microbiological controls and
control points in a hake fillets manufacturing process for exportation.
Arch Latinoam Nutr. 50(2), 171–176.
D’AOUST, J.Y., GELINAS, R. and MAISHMENT, C. 1980. Presence of
indicator organisms and recovery of Salmonella in fish and shellfish. J.
Food Prot. 43, 679–682.
 MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS 39

DAVIES, A.R., CAPELL, C., JEHANNO, D., NYCHAS, G.J.E. and

KIRBY, R.M. 2001. Incidence of foodborne pathogens in European
fish. Food Control 2, 67–71.
DESMARCHELIER, P.M. 1997. Pathogenic Vibrios. In Foodborne Microor-
ganisms of Public Health Significance, 5th Ed. (A.D. Hocking, G. Arnold,
I. Jenson, K. Newton and P. Sutherland, eds.) pp 287–312, AIFST NSW
Branch, Food Microbiology Group, North Sydney, Australia.
DILLON, R., PATEL, T. and RATMAN, S. 1992. Prevalence of Listeria in
smoked fish. J. Food Prot. 55, 866–870.
DILLON, R., PATEL, T. and RATMAN, S. 1994. Occurence of Listeria in
hot and cold smoked seafood products. Int. J. Food Microbiol. 22, 73–
77.
FARBER, J. 1991. Listeria monocytogenes in fish products. J. Food Prot. 54,
922–924, 934.
FENWICK, S.G. and MCCARTHY, M.D. 1995. Yersinia enterocolitica is a
common cause of gastroenteritis in Auckland. N. Z. Med. J. 108, 269–
271.
FRICKER, C.R. and TOMPSETT, S. 1989. Aeromonas spp in foods: A
significant cause of food poisoning? Int. J. Food Microbiol. 9, 17–23.
GENNARI, M., DRAGOTTO, F. 1992. A study of the incidence of different
fluorescent Pseudomonas species and biovars in the microflora of fresh
and spoiled meat and fish, raw milk, cheese, soil and water. J. Appl.
Bacteriol. 72(4), 281–288.
GILLESPIE, I.A., ADAK, G.K., O’BRIEN, S.J., BRETT, M.M., BOLTON,
F.J. 2001. General outbreaks of infectious intestinal disease associated
with fish and shellfish, England and Wales, 1992–1999. Commun. Dis.
Public Health 4(2), 117–123.
GOBAT, P. and JEMMI, T. 1993. Distribution of mesophilic Aeromonas
species in raw and ready to eat fish and meat products in Switzerland.
Int. J. Food Microbiol. 20, 117–120.
GRAM, L. 1993. Inhibitory effect against pathogenic and spoilage bacteria
of Pseudomonas strains isolated from spoiled and fresh fish. Appl
Environ Microbiol 59(7), 2197–2203.
HANNINEN, M.L., OIVANEN, P. and HIRVELA-KOSKI, V. 1997.
Aeromo-nas species in fish, fish eggs, shrimp and freshwater. Int. J.
Food Micro-biol. 34, 17–26.
HEINITZ, M.L., RUBLE, R.D., WAGNER, D.E. and TATINI, S.R. 2000.
Incidence of Salmonella in fish and seafood. J. Food Prot. 63(5), 579–
592.
HOOK, D., JALALUDIN, B. and FITZSIMMONS, G. 1996. Clostridium
perfringens foodborne outbreak: An epidemiological investigation.
Aust. N. Z. J. Public Health 20, 119–122.
40 C. PAPADOPOULOU ET AL.

HUDSON, J.A., MOTT, S.J., DELACY, K.M. and EDRIDGE, A.L. 1992.
Incidence and coincidence of Listeria spp., motile aeromonads and
Yers-inia enterocolitica on ready to eat fleshfoods. Int. J. Food
Microbiol. 16, 99–108.
ISONHOOD, J.H. and DRAKE, M. 2002. Aeromonas species in foods.
J. Food Prot. 65(3), 575–582.
JAY, S., GRAU F.H., SMITH, K., LIGHTFOOT D., MURRAY, C.,
DAVEY, G. 1997. Salmonella In Foodborne Microorganisms of Public
Health Significance, 5th Ed. (A.D. Hocking, G. Arnold, I. Jenson, K.
Newton and P. Sutherland, eds.) pp 169–229, AIFST NSW Branch,
Food Microbiol-ogy Group, North Sydney, Australia.
JENSEN, G.L. and GREENLEES, K.J. 1997. Public health issues in aquac-
ulture. Rev. Sci. Tech. 16(2), 641–51.
JONSSON, I., MONSEN, T., WISTROM, J. 1997. A case of Plesiomonas
shigelloides cellulitis and bacteraemia from northern Europe. Scand. J.
Infect. Dis. 29(6):631–632.
KHARE, S.S., KAMAT, A.S., DOCTOR, T.R. and NAIR, P.M. 1996. Inci-
dence of Yersinia enterocolitica and related species in some fish, meat
and meat products in India. J. Sci. Food Agric. 72, 187–195.
KIROV, M.S. 1997. Aeromonas. In Foodborne Microorganisms of Public
Health Significance, 5th Ed. (A.D. Hocking, G. Arnold, I. Jenson, K.
Newton and P. Sutherland, eds) pp 473–492, AIFST NSW Branch,
Food Microbiology Group, North Sydney, Australia.
KNEBEL, U., SLOOT, N., EIKENBERG, M., BORSDORF, H., HOFFLER,
U. and RIEMANN, J.F. 2001. Gastroenteritis due to Plesiomonas
shigelloides – rare cases in the Western world. Med. Klin. 96(2), 109–
113.
LACIAR, A.L. and DE CENTORBI, O.N.P. 2002. Listeria species in
seafood: isolation and characterization of Listeria spp. from seafood in
San Luis, Argentina. Food Microbiol. 19, 645–651.
LANSER, J., ORMEROD, S. and BARTON, M. 1993. Yersinia enterocolitica:
Another enteric pathogen on the increase. IMVS Newsletter 22, 7–8.
LIPP, E.K. and ROSE, J.B. 1997. The role of seafood in foodborne diseases
in the United States of America. Rev. Sci. Tech. 16(2), 620–640.
PAPADOPOULOU, C., DIMITRIOU, D., LEVIDIOTOU, S., PANAGIOU,
A., SAVAIDIS, J., GOLEGOU, S., ANTONIADES, G. 1996. Microbial
flora of fresh and marine fish. In Abstract Book, 1st Public Health and
Health Services Panhellenic Congress, 18–20 March (J. Kourea-
Kremastinou, Chairman) p 69, National School of Public Health,
Athens, Greece.
PERRIN, M., BEMER, M. and DELAMARE, C. 2003. Fatal case of Listeria
innocua bacteremia. J. Clin. Microbiol. 41(11), 5308–5309.
 MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS 41

POLLOCK, A.M. and WHITTY, P.M. 1991. Outbreak of Clostridium

perfrin-gens food poisoning. J. Hosp. Infect. 17, 179–186.
RAMOS, M. and LYON, W.J. 2000. Reduction of endogenous bacteria asso-
ciated with catfish fillets using the Grovac process. J. Food Prot. 63(9),
1231–1239.
REILLY, P.J.A. and TWIDDY, D.R. 1992. Salmonella and Vibrio cholerae
in brackish water cultured tropical prawns. Int. J. Food Microbiol. 16,
293– 301.
ROBERTS, D., HOOPER, W., GREENWOOD, M. 1995. Practical Food
Microbiology, 2nd Ed. PHLS, Colindale-London, UK.
ROCOURT, J., JACQUET, C. and REILLY, A. 2000. Epidemiology of
human listeriosis and seafoods. Int J Food Microbiol. 62(3), 197–209.
RODRIGUEZ-JEREZ, J.J., LOPEZ-SABATER, E.I., ROIG-SAGUES, A.X.
and MORA-VENTURA, M.T. 1994. Histamine, cadaverine and
putrescine forming bacteria from ripened Spanish semipreserved ancho-
vies. J. Food Sci. 59(5), 998–1001.
RORVIK, L.M., CAUGANT, D.A. and YNDESTAD, M. 1995. Contamina-
tion pattern of Listeria monocytogenes and other Listeria spp. in a
salmon slaughterhouse and smoked salmon processing plant. Int. J.
Food Micro-biol. 25, 19–27.
SMOLOWITZ, R., WADMAN, E. and CHIKARMANE, H.M. 1998.
Pseudomonas putida infections of the oyster toadfish (Opsanus tau). Biol.
Bull. 195(2), 229–231.
TAYLOR, S. 1986. Histamine food poisoning: Toxicology and clinical
aspects. CRC Crit. Rev. Toxicol. 17(2), 91–128.
TRYFINOPOULOU, P., TSAKALIDOU, E. and NYCHAS, G.J. 2002.
Char-acterization of Pseudomonas spp. associated with spoilage of gilt-
head sea bream stored under various conditions. Appl Environ
Microbiol 68(1), 65–72.
TSAI, G.J. and CHEN, T.H. 1996. Incidence and toxicity of Aeromonas
hydrophila in seafood. Int. J. Food Microbiol. 31, 121–131.
VAN DAMME, L.R. and VANDEPITTE, J. 1980. Frequent isolation of
Edwardsiella tarda and Pleisiomonas shigelloides from healthy Zairese
freshwater fish: A possible source of sporadic diarrhea in the tropics.
Appl. Environ. Microbiol. 39(3), 475–479.
VANDEPITTE, J., VAN DAMME, L., FOFANA, Y. and DESMYTER, J.
1980. Edwardsiella tarda and Plesiomonas shigelloides. Their role as
diarrhea agents and their epidemiology. Bull. Soc. Pathol. Exot. Filiales
73(2), 139–149.
VAZ-VELHO, M., DUARTE, G. and GIBBS, P. 1998. Occurrence of
Listeria spp. in salmon trout (Oncorhynchus mykiss) and salmon (Salmo
salar). Food Sci. Technol. Int. 4, 121–125.
42 C. PAPADOPOULOU ET AL.

VAZ-VELHO, M., DUARTE, G. and GIBBS, P. 2000. Evaluation of mini-

VIDAS rapid test for detection of Listeria monocytogenes from
production lines of fresh to cold-smoked fish. J. Microbiol. Methods 40,
147–151.
VAZQUEZ-JUAREZ, R., ANDLID, T. and GUSTAFSSON, L. 1994. Cell
surface hydrophobicity and its relation to adhesion of yeasts isolated
from fish gut. Colloid. Surface. B: Bionterfaces 2, 199–208.
VELAZQUEZ, L.D.C., ESCUDERO, M.E. and GUZMAN, A.M.S. 1996.
Prevalence of Yersinia enterocolitica in Hake (Merluccius hubbsi) fillets.
J. Food Prot. 59, 781–783.
WALKER, S.J. and BROOKS, J. 1993. Survey of the incidence of
Aeromonas and Yersinia species in retail foods. Food Control 4, 34–40.
WALLACE, B.J., GUZEWICH, J.J., CAMBRIDGE, M., ALTEKRUSE, S.
and MORSE, D.L. 1999. Seafood-associated disease outbreaks in New
York, 1980–1994. Am. J. Prev. Med. 17(1), 48–54.
YASUMOTO, T. 2000. Historic considerations regarding seafood safety. In
Seafood and Freshwater Toxins (L.M. Botana, ed.) pp. 1–17, Marcel
Dekker Inc, New York, NY.
YOUSSEF, H., EL TIMAWY, A.K. and AHMED, S. 1992. Role of aerobic
intestinal pathogens of fresh water fish in transmission of human dis-
eases. J. Food Prot. 55, 739–740.