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BACTERIAL FOOD INTOXICATION

Santosh Kumar Singh1, Vidya Dhar Pandey2* and Vijay Chandra Verma1

Abstract
A wide variety of foods and food products derived from plants and animals
support the growth of pathogenic and toxigenic bacteria, resulting in foodborne diseases such as food infection and food intoxication or poisoning
which is a major public health problem globally. Bacterial growth and toxin
production in foods are influenced by various intrinsic (e.g. pH, moisture,
redox potential, nutrients) and extrinsic (e.g. temperature) factors. Generally,
food poisoning outbreaks occur as the result of poor food hygiene, and
improper handling, storage and preservation of foods and food products.
Bacterial food intoxication is a food-borne illness caused by the ingestion
of food containing preformed bacterial toxins which are produced as a result
of bacterial growth in the food. The important bacterial species that are most
commonly involved in food poisoning outbreaks are Clostridium botulinum,
Clostridium perfringens, Staphylococcus aureus and Bacillus cerus. Their
toxins mainly act on digestive or nervous systems, leading to severe disorders
and sometimes death. Food poisoning or intoxication can be prevented by
adopting good hygienic practice with proper methods and conditions of food
handling, storage and preservation. Various sensitive and effective
techniques and methods are now available for the detection of food-borne
pathogenic microbes and their toxins.
Key words: Microbial toxin, enterotoxins, botulism, food intoxication,
neurotoxins
1. Centre of Experimental Medicine and Surgery (CEMS), Institute of Medical Sciences,
Banaras Hindu University, Varanasi-221005, Uttar Pradesh
2. Department of Botany, Government Post-Graduate College, Rishikesh-249201, Dehradun,
Uttarakhand
*

Corresponding author: E-mail: vdpandey2@yahoo.co.in

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1. INTRODUCTION
Microorganisms present in foods may cause food spoilage or food-borne
diseases. Various human foods support the growth of pathogenic
microorganisms or act as their carriers for transmission. Food-borne diseases
are a major public health problem globally. Numerous outbreaks of foodborne diseases in various countries have been reported. A food-borne disease
outbreak refers to the occurrence of two or more cases of similar illness
resulting from the ingestion of a common food (Centers for Disease Control
and Prevention, 1996). In USA alone, an estimated 25-81 million cases of
food-borne illness and 9000 deaths are known to occur annually (GernerSmidt and Whichard, 2007). Number of annual cases of food-borne illness
in Australia is about 5.4 million. In France, 75000 cases of food-borne illness
are estimated every year, while in Japan there were 9966 cases per year.
Unfortunately, reliable comparable figures from the under-developed and
developing countries are not available (Granum, 2006; Farthing and Kelly,
2007; IASR, 2008; Hamer and Gorbach 2008; Acheson, 2009; Senior, 2009),
but one can assume that poor hygienic conditions and poor sanitation in
these countries would favour the greater prevalence of food-borne diseases.
In India, food-borne diseases are rarely recorded. Due to high prevalence
of other severe diseases in under-developed and developing countries, foodborne diseases are considered of minor importance. Food-borne illness is
caused by pathogenic organisms such as bacteria, viruses and parasites or
their toxins present in foods. More than 250 food-borne diseases are known.
With the exception certain parasites, food-borne pathogens are microscopic
in size. Food-borne illness caused by microorganisms is generally classified
in to two categories: food infection (food-borne infection) and food intoxication
(food-borne intoxication). The term food poisoning is often used very loosely
and to include food-borne diseases resulting from both food infection and
food intoxication. But in true and strict sense it refers to the diseases caused
by food intoxication.
Bacteria constitute an important group of food-borne pathogens which
are implicated in various food-borne diseases, resulting from both food
infection and intoxication. Bacterial food infection refers to food-borne illness
caused by ingestion of food containing viable (live) bacteria (e.g. Salmonella,
Listeria, pathogenic E. coli) which then grow or multiply in the host, leading
to illness. Bacterial food intoxication, on the other hand, refers to foodborne illness caused by the ingestion of food containing preformed bacterial
toxins (e.g. toxins produced by Staphylococcus aureus and Clostridium

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botulinum) which are produced as a result of bacterial growth in the food.

In addition to food infection and intoxication, the third type of bacterial
food-borne disease is food toxicoinfection which occurs when bacteria (e.g.
Clostridium perfringens) present in food are ingested that subsequently
produce toxins in the host. On the basis of roles of foods, food infections can
be of two types: (i) those in which food merely acts as the carrier of pathogens
and does not support the growth or multiplication of pathogens, e.g. bacteria
causing tuberculosis, typhoid fever, diphtheria, cholera, brucellosis,
infectious hepatitis, Q fever etc., and (ii) those in which food serves as a
culture medium for growth of pathogens, e.g. Salmonella sp.,
enteropathogenic Escherichia coli, Vibrio parahaemolyticus etc. In general,
food intoxication is manifested more rapidly than food infections. Symptoms
of food-borne illness range from mild discomfort to severe reactions, and
may even result in death. The general symptoms include nausea, vomiting,
headache, abdominal cramps, diarrhoea, bloody diarrhoea and fever. The
severity of the illness depends on various factors which include amount of
contaminant ingested and susceptibility to food-borne illness. Babies and
young children, pregnant women, old persons, AIDS patients, persons with
impaired immune systems and those taking immunosuppressive drugs are
more susceptible. Usually, the incubation period of food intoxication is very
short, 4-24 hours. In contrast, the incubation period of food infection is
generally 12-72 hours, although it may be longer in some cases. The most
commonly involved bacteria in food intoxication or poisoning are
Clostridium botulinum, Clostridium perfringens, Staphylococcus aureus
and Bacillus cereus. Presently, many techniques and methods are available
for enumeration and isolation of microbial contaminants in food, but those
given by American Public Health Association (APHA, 1984) and
International Commission on Microbiological Specification of Foods (ICMSF,
1978) are widely acceptable. Moreover, several novel immunological,
chemical, biochemical and biophysical methods are in use to monitor and
detect food-borne pathogenic microbes and their toxins in foods and food
products. Some relevant and sensitive techniques include the use of nucleic
acid probes, polymerase chain reaction (PCR) and biosensors (Fung, 1995).

2. FACTORS AFFECTING BACTERIAL GROWTH IN FOODS

Various foods consumed by humans are good substrates for growth and
survival of both pathogenic and non-pathogenic microorganisms, including
bacteria. Foods of animal and plant origin are complex biological materials

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which vary in nutrient content and other properties. The growth and
survival of microorganisms in human foods are influenced by both extrinsic
(e.g. temperature) and intrinsic (e.g. pH, moisture, redox potential,
nutrients) factors.

2.1. Temperature
Every microorganism has a minimal, a maximal and an optimal temperature
for growth. On the basis of temperature ranges for growth, bacteria can be
grouped as: thermophiles (optimum temperature 55-75 oC), mesophiles
(optimum temperature 30-45 oC), psychrophiles (optimum temperature 515 oC) and psychrotrophs (optimum temperature 25-30oC). Generally, foodspoilage and pathogenic bacteria are mesophiles. However, some food-borne
pathogenic bacteria (e.g. Listeria monocytogenes, Yersinia enterocolitica)
are psychrotrophic which are responsible for disease outbreaks due to the
consumption of food products stored at refrigeration temperature. Decrease
and increase in temperature below and above the optimum growth range
causes the inhibition of bacterial growth in foods. The resistant spores
produced by many bacteria are known to survive both freezing and heating.

2.2. pH
Like other microorganisms, bacteria have minimum, optimum and maximum
pH for growth. Bacterial growth is significantly affected by the pH of foods.
Most foods are neutral or acidic. In general, fungi (yeasts and molds) are
more acid-tolerant than bacteria. Most bacteria are unable to grow below
pH 3.5 and those implicated in food-borne diseases are unable to grow below
pH 4.5.

2.3. Moisture
The growth of bacteria and other microorganisms in or on food depends on
the available water which is best expressed in terms of water activity (aw).
The water activity of a food or solution is the ratio of the water vapour
pressure of the food or solution (p) to that of pure water (p0) at the same
temperature. Water activity of pure water is assumed to be 1. The value of
aw of a solution decreases below 1 with the increase in concentration of the
solution. Foods exhibit significant variation in aw. Most bacteria and fungi
are unable to grow in foods having aw less than 0.90.

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2.4. Redox Potential

The redox potential, also called oxidation-reduction potential, of a biological
system is an index of degree of its oxidation. It is denoted as Eh and measured
in terms of millivolts (mV). The redox potential of foods is an important
selective factor which determines the type of organisms which will grow.
The redox potential of a food is related to its chemical constituents and the
oxygen partial pressure. Bacteria are classified as aerobic, anaerobic,
facultative or micro-aerobic based on the Eh required for their growth. Eh
value ranges from about +300 mV for aerobic bacteria to 420 mV for
anaerobic bacteria.

2.5. Nutrients
Nutritional quality and quantity of a food determines the type of bacteria
that are likely to be present in the food. The nutritional requirements of
bacteria may change depending on the environmental conditions. In
general, most human foods provide adequate nutrients for most
microorganisms. Foods rich in proteins favour the growth of bacteria with
complex nutritional needs. Starchy foods favour the growth of bacteria that
are able to break down the complex carbohydrates. Bacteria producing
lipolytic enzymes are favoured in lipid-containing foods.

3. BACTERIAL FOOD INTOXICATION

3.1. Botulism
Botulism, a neuroparalytic disease paralyzing or weakening skeletal muscle
in the body, is caused by the ingestion of food containing toxin produced by
the bacterium Clostridium botulinum. It is associated with the consumption
of poorly preserved and improperly handled foods. C. botulinum is a grampositive, rod shaped, spore-forming and anaerobic bacterium that is
commonly found in soils, aquatic sediments, rotting vegetation and digestive
tracts of animals and birds. It is a common contaminant of fishes, occurring
in their intestinal tracts. Many vegetables and fruits can easily become
contaminated with spores of bacterium because they are often in contact
with soil. The toxin (an exotoxin) produced by the bacterium is a protein
neurotoxin, called botulinum neurotoxin (BoNT) or simply botulinum toxin,
which is most potent toxic substance in nature. There are four types of
botulism: (i) food-borne botulism, an intoxication caused by ingestion of

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food containing preformed BoNT, (ii) wound botulism, which occurs due to
the production of BoNT in vivo after growth of C. botulinum in an infected
wound, (iii) infant botulism, which results from the production of BoNT in
vivo in the intestinal tract of an infant colonized with C. botulinum, and
(iv) botulism due to the intestinal colonization in children older than infants
and in adults in which no food or wound source is implicated (Hatheway
and Johnson, 1998). There are seven immunologically or antigenically
distinct types of BoNT which are designated as A, B, C, D, E, F and G.
Types A (BoNT/A), B (BoNT/B), E (BoNT/E) and F(BoNT/F) are the principal
causes of human botulism. Among these BoNT/A is most dangerous and is
responsible for most outbreaks of food-borne botulism. Types C (BoNT/C)
and D (BoNT/D) are associated with botulism in birds and mammals. Type
G (BoNT/G) has not been clearly implicated in botulism (Hatheway, 1998).
Irrespective of type, each BoNT is synthesized as a single-chain inactive
polypeptide (150 kDa) that subsequently undergoes proteolytic cleavage,
resulting in the generation of active form of the toxin which consists of two
polypeptide chains linked by a single disulfide bond. The two chains of the
toxin are called light chain (50 kDa) and heavy chain (100 kDa). The toxin
is released by the lysis of bacterial cells. Toxin production depends on the
ability of the bacterial cells to grow in food and to autolyse. Factors such as
composition of the food, temperature of storage and pH of food influence
spore germination, growth and toxin production. Conditions that favour C.
botulinum growth and toxin production include a relatively high moisture,
low salt concentration, low acidic pH (pH >4.6), anaerobic condition and
food stored without refrigeration. The main sources of botulism are canned
meat, fish, string beans, sweet corn, beets, and other low and medium acidic
foods. Spores of C. botulinum can survive long storage periods in raw and
precooked frozen foods and can germinate when conditions become ideal.
Botulinum toxins can be destroyed by heat treatment. However, the
temperature and duration of treatment depend on the type of toxin.
After absorption in the small intestine, the toxin is transported in to
lymphatic channels and then in to bloodstream. The toxin reaches the
cholinergic synapses of peripheral nervous system by blood circulation where
it acts. The botulinum neurotoxins are extremely potent. They inhibit the
release of the neurotransmitter acetylcholine at the neuromuscular junction,
resulting in flaccid muscle paralysis. Their action involves three sequential
steps (Simpson, 1981). In the first step, the toxin binds rapidly and
irreversibly to receptors on the presynaptic nerve surface (Kozaki, 1979;
Murayama et al., 1984). In the second step, which is an internalization

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stage, the toxin crosses the plasmalemma and enters in to the nerve
terminus. It appears that the heavy-chain moiety of the toxin is capable of
inducing channel formation in membranes at low pH (Shone et al., 1987).
It is not known whether the whole toxin or a fragment(s) thereof is/are
delivered to the cytosol by the internalization process, although recent work
suggests that internalization of both the heavy and light chains of
neurotoxin may be required for the inhibition of neurotransmitter release
(Poulain et al., 1989). In the third step, the toxin exerts its action by
inhibiting neurotransmitter release. Normally, the arrival of an action
potential at the nerve ending triggers an influx of calcium ions which itself
promotes the exocytosis of acetylcholine from vesicles at active zones on the
plasmalemma. Several possible mechanisms of inhibition have been proposed
by various workers (Molgo and Thesleff 1984; Sanchez-Prieto et al., 1987;
Shone and Hambleton 1989; Simpson, 1981). The toxins may alter the rate
of calcium ion efflux and influx or act directly on a component of the
acetylcholine release mechanism such as a vesicle protein or cytoskeleton
component.
Typical symptoms of botulism usually occur within 12-36 hours after
consumption of the contaminated food. Early symptoms are digestive
disturbances followed by nausea, vomiting, diarrhoea, together with
dizziness and headache. Double vision may occur early and there may be
difficulty in speaking. Mouth may become dry, throat constricted and tongue
may get swollen and coated. Involuntary muscles become paralysed and
paralysis spreads to the respiratory system and to the heart. Death normally
results from respiratory failure. In fatal cases, death occurs within 3-6 days
after the poisonous food has been ingested. Information regarding the
outbreaks and occurrence of botulism is available for few countries. In USA,
160 outbreaks of food-borne botulism were reported during 1990 to 2000
and food items involved in these cases were home-canned (Sobel et al.,
2004). A total 91 cases of botulism were reported in Alaska alone between
1990 and 2000. Botulism among Alaska natives was associated with the
eating of whale meat. Adoption of food hygiene and increased awareness
about disease resulted in decreased mortality rate in natives in later decades
(Chiou et al., 2002; Eisenberg and Bender, 1976). In United Kingdom, an
outbreak of 27 cases was reported in 1989 which was linked to the
consumption of commercial hazelnut yogurt. Botulism is a major public
health problem in Argentina, which is due to consumption of improperly
processed meats and vegetables as well as home-canned food. Between 1992
and 2004, forty-one food-borne botulism cases were reported (Tornese et
al., 2008).

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The quick detection of botulism is needed for the timely administration

of anti-toxin. Amplified enzyme-linked immunosorbent assay (ELISA) and
amplified ELISA/enzyme-linked coagulation assay (ELCA) shows promise
for rapid and sensitive detection of toxin (Ferreira, 2001; Ferreira et al.,
2003; Roman et al., 1994). The methods of botulism prevention include: (i)
use of approved heat processes for canned foods, (ii) rejection of all spoilt
canned foods, (iii) avoiding food that has been semi-cooked and not well
heated, and (iv) boiling of suspected food for at least 15 minutes to denature
the toxin. The only known method of successful treatment of botulism is by
the administration of anti-toxin.

3.2. Staphylococcal Intoxication

Staphylococcal food poisoning (SFP) or intoxication is one of the common
types of food-borne disease which is caused by the ingestion of foods
containing the thermostable enterotoxins produced by certain strains of
the bacterium Staphylococcus aureus. S. aureus is a facultative anaerobe,
gram-positive coccus, non-motile, and catalase and coagulase positive. It is
widely distributed in air, dust, sewage, water, foods, humans and animals.
Humans and animals are the primary reservoirs for this microbe. It is present
as normal microflora of human beings and colonizes skin. But, it can cause
various infections and diseases such as boils, carbuncles, bullous impetigo,
toxic shock syndrome, scalded skin syndrome, enterocolitis, osteomyelitis
and food poisoning. Although food handlers are usually the main source of
food contamination in food poisoning outbreaks, equipments used in food
handling and environmental surfaces can also be sources of contamination
with S. aureus. It is usually transferred to the foods and food products due
to their poor handling. The growth of the bacterium is favoured by proteinrich foods with high salt content. Foods that are frequently incriminated in
staphylococcal food intoxication include meat and meat products, poultry
and egg products, fish and fish products, salads, milk and milk products,
and cream-filled bakery products. If the contaminated food is stored at
temperature that encourages the growth of these organisms, production of
enterotoxins occurs in the food and after ingestion of this food it will spread
rapidly in humans. Staphylococcus aureus is able to grow in a wide range
of temperatures (7 to 48.5C with an optimum of 30 to 37C), pH (4.2 to 9.3
with an optimum of 7 to 7.5) and sodium chloride concentrations up to 15%
(Schmitt et al., 1990).

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The appearance of symptoms of food poisoning depends upon quantity,

type and toxicity of the toxin. Bacterial population greater than 105 cells
per gram of food are needed to produce sufficient amounts of staphylococcal
enterotoxins to elicit symptoms of food intoxication. In contrast to other
food-borne diseases, which have longer incubation periods, onset of
staphylococcal intoxication mostly occurs within 2-4 hours following ingestion
of toxin-contaminated foods (Bergdoll, 1989). Common symptoms include
abdominal cramps, nausea, vomiting, diarrhoea, dehydration, prostration,
weakness and subnormal body temperature. Staphylococcal food
intoxication is rarely fatal and recovery usually occurs within 24-48 hours.
Staphylococcal enterotoxin (SE) is thermostable exotoxin which acts upon
the gastrointestinal epithelium. They are single chain, low-molecular weight
proteins. The toxins are resistant to proteolytic enzymes such as trypsin
and rennin and thus keep their activity in the digestive tract after ingestion.
Cooking may kill the bacterial cells but the toxin may still remain active
due to their heat stability. Although purified toxins become rapidly
inactivated by heat, they are much more heat resistant when they are in
foods. Several antigenically distinct types of staphylococcal enterotoxins
are produced by strains of S. aureus which are named as A, B, C1, C2, C3,
D, E, G, H, I and J (Balaban and Rasooly, 2000). Among these,
staphylococcal enterotoxin A is the most toxic and implicated in most of the
staphylococcal food intoxication. In addition to enterotoxins, which are
implicated in food intoxication, S. aureus produces other toxins such as
hemolysins, leukocidins, exfoliative toxins and toxic syndrome toxins which
are known to be associated with different diseases in humans.
Numerous sensitive methods are now available for the detection of
enterotoxins in food products. Enzyme-linked immunosorbent assay (ELISA)
and reversed passive latex agglutination (RPLA) method can detect
nanogram of enterotoxins in one gram or in one milliliter of food products
(Sharma et al., 2000; Burdova et al., 1994; Gouloumes et al., 1996; Strachan
et al., 1997). DNA amplification methods like polymerase chain reaction
(PCR) can show the presence of enterotoxigenic strains of Staphylococcus
aureus before the expression of enterotoxins on the basis of specific gene
sequences and in this way it is helpful to take preventive majors before
release of toxins (Tsen and Chen 1992; Holeckova et al., 2002). Biosensors
are currently being developed to provide real-time detection of staphylococcal
enterotoxins (Nedelkov et al., 2000).

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Regulation of the temperature is one of the most effective measures to

control staphylococcal growth as well as food intoxication. Inadequate
cooling and refrigeration of food is the main cause of food poisoning outbreak
in the majority of cases. Therefore, adequate heat processing and cooking
and proper cooling and refrigeration are important preventive measures.
Staphylococcus aureus causes food intoxication or poisoning in various
countries of the world (Bergdoll 1989). In India, rate of food intoxication is
very high because of warm and humid climate which favours the growth of
the bacterium. Epidemic outbreaks of staphylococcal food poisoning (SFP)
are relatively rare in developed countries. SFP accounts for only 1.3% of
the total estimated cases of food-borne illnesses caused by known agents in
USA (Holmberg and Blake, 1984; Mead et al., 1999; Wieneke, 1993). In
the early 1980s, SFP was reported to account for 14% of total food borne
outbreaks in USA. Similar decrease in frequency has been reported in
Japan. Before 1984, 25-35% of all cases of bacterial food- borne illness in
Japan involved SFP, whereas in the late 1990s, only 2-5% of incidents
involved SFP. The foods that are most often involved in staphylococcal food
poisoning differ widely among countries. This may be due to differences in
the consumption and food habits in different countries.

3.3. Bacillus cereus Intoxication

Bacillus cereus is a gram-positive, spore-forming, motile, aerobic, rod-shaped
bacterium that is commonly found in soil and on vegetation. In addition to
food poisoning, it causes opportunistic infections. It easily spreads and
colonizes a wide range of foods and food products of both plant and animal
origin (e.g. meat, eggs and dairy products) (Kramer and Gilbert, 1989).
Due to the significant difference in the amount of enterotoxin produced by
different strains of B. cereus, the total infective dose seems to vary between
about 105 and 108 viable cells or spores (Granum, 1997). Spores of B. cereus
generally survive cooking. B. cereus is known to produce one emetic toxin
and three different enterotoxins. Two types of B. cereus food poisoning have
been reported. The first type, caused by an emetic toxin, produces nausea
and vomiting, while the second type, caused by enterotoxins, results in
diarrhea (Agata, 1994). The diarrhoeal type of food-poisoning is characterized
by abdominal cramps and pain with diarrhea. In a small number of cases
both types of symptoms are recorded (Kramer and Gilbert, 1989), probably
due to production of both types of toxins. The diarrhoeal type of food
poisoning is caused by complex enterotoxins produced during vegetative

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growth of B. cereus in the small intestine (Beecher, 1997; Lund and Granum,
1997; Granum, 1994), while the emetic toxin is produced by growing cells
in the food (Kramer and Gilbert, 1989). The emetic toxin has been named
cereulide and consists of a ring structure of three repeats of four amino
and/or oxy acids: [D-O-Leu-D-Ala-L-O-Val-L-Val]3. This ring structure has a
molecular mass of 1.2 kDa and is chemically analogue to the potassium
ionophore valinomycin. The emetic toxin is resistant to heat, pH and
proteolysis.
Among the three different enterotoxins of B. cereus, two have been
shown to be involved in food poisoning (Agata, 1995; Beecher, 1994, 1995;
Lund and Granum, 1996). B. cereus produces different components of the
enterotoxins (Beecher, 1994, 1995, 1997; Lund and Granum, 1996, 1997).
A three-component hemolysin (HBL) consisting of three proteins (B, L1
and L2) with enterotoxin activity have been reported (Beecher, 1994, 1995,
1997). A non-hemolytic three-component enterotoxin (NHE) was also
identified (Lund and Granum, 1996). The three components of this toxin
were different from the components of HBL. One enterotoxin protein had a
molecular mass of about 57 kDa and another about 100 kDa (Kramer and
Gilbert, 1989). The largest could be the 105 kDa component of the NHE.
The third enterotoxin T is a single component protein, but has not been
shown to be involved in food poisoning. It has been suggested, from studies
of interactions with erythrocytes, that the B protein is the component that
binds HBL to the target cells, and that L1 and L2 have lytic functions
(Beecher and Macmillan, 1991). Another proposed model for the action of
HBL, suggests that the components of HBL bind to target cells independently
and then constitute a membrane-attacking complex resulting in a colloid
osmotic lysis (Beecher, 1997). Studies of interactions between NHE and
Vero cells have shown that a protein of the 105 kD might be the binding
component of that complex (Lund and Granum, 1997). The two other
components are probably not able to bind to these cells alone.
Temperature is one of the most important factors that can be controlled
to reduce or eliminate B. cereus growth as well as food poisoning. The
temperature required to prevent the growth of B. cereus in food is greater
than 60C (140F) until served, or rapidly cooling foods to below 10C (50F)
for storage. Ideally, foods should be cooled to below 15C (59F) within two
to three hour after cooking. Disease caused by B. cereus differs from country
to country. In Japan the emetic type is reported about 10 times more
frequently than the diarrhoeal type, while in Europe and North America

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the diarrhoeal type is the most frequently reported (Kramer and Gilbert,
1989). B. cereus food poisoning has been reported in relatively few countries
of Europe. B. cereus is the cause of 33% of the total cases of food poisoning
(excluding virus) in Norway (1988-1993) (Granum, 1994), 47% in Iceland
(1985-1992), 22% in Finland (1992), 8.5% in The Netherlands (1991), and
5% in Denmark (1990-92) (Schmidt, 1995). Much lower numbers have been
reported from other countries, such as England and Wales (0.7%), Japan
(0.8%), USA (1.3%) and Canada (2.2%) (Kramer and Gilbert, 1989).

3.4. Clostridium perfringens Intoxication

Clostridium perfringens is a gram-positive anaerobic bacterium which is
widely distributed in soil, sewage, and intestinal tracts of humans and
animals. It causes gas gangrene, a disease characterized by muscle necrosis
and invasion of the bloodstream. Additionally, it causes food poisoning
resulting from the ingestion of foods containing its viable vegetative cells
which sporulate in the intestine and produces an enterotoxin. In the small
intestine, the enterotoxin binds to receptors on the cell surface of intestinal
epithelial cells and causes damage to cell membranes leading to alteration
in membrane permeability. It can produce more than 13 toxins, among
which C. perfringens enterotoxin (CPE) is responsible for type A food
poisoning outbreaks (Hatheway, 1990; Labbe, 1991; Milton et al., 2005).
The bacterium is most frequently associated with meat and poultry products.
The symptoms include predominantly diarrhoea, vomiting and abdominal
pain. The incubation period is 24-30 hours. Death occurs occasionally among
debilitated patients, particularly the old and already diseased patients
(Labbe, 1989). Unlike other toxins, CPE is produced only by sporulating
cells and accumulates in a large inclusion body inside the mother cell, from
which it is released after lysis at the end of sporulation (Loffler and Labbe,
1985, 1986). Based on the presence of four major lethal toxins (alpha-,
beta-, epsilon-, and iota-toxins), C. perfringens is classified into five toxigenic
types (A to E), and each type can cause different diseases. The most commonly
encountered is type A (alpha-toxin) which causes gas gangrene
(myonecrosis), diarrhoea, and food poisoning in humans (Hatheway, 1990).
Type B (alpha-, beta- and epsilon-toxins) and type D (alpha- and epsilon
toxins) toxins are the causative agents of fatal enterotoxemia in domestic
animals and occasionally humans (Yamaghishi et al., 1997). The type C
(alpha- and beta-toxins) toxin causes severe necrotic enteritis in humans
and is called Darmbrand in Germany and pig-bel in New Guinea (Petrillo

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et al., 2000). The pathogenicity of type E (alpha- and iota-toxins) strains is

unclear and has rarely been isolated from humans (Yamaghishi et al., 1997).
Ingestion of food containing approximately 108 cells of enterotoxigenic
strain of C. perfringens causes food poisoning (Shandera et al., 1983; Fach
and Popoof, 1997). In vivo production of the enterotoxin is associated with
sporulation in the intestine (Collie et al., 1998), while in vitro production is
obtained in appropriate culture media (Tortora, 1984; Tortora et al., 1986).
CPE action involves formation of a series of complexes in mammalian plasma
membranes. One such CPE-containing complex of about 155 kDa is
important for the induction of plasma membrane permeability alterations.
These membrane permeability changes cause damage in epithelium,
allowing the enterotoxin to interact with the tight junction (TJ) protein
occludin. CPE-occludin interactions result in formation of a 200 kDa CPE
complex and facilitate internalization of occludin into the cytoplasm. The
occludin (and possibly other proteins) damages TJs and disrupts the normal
paracellular permeability barrier of the intestinal epithelium, which may
contribute to CPE-induced diarrhea (McClane and Chakrabarti, 2004).
Enterotoxigenic strains can be characterized by the presence of the cpe
gene by means of molecular methods such as PCR, ELISA, reverse passive
latex agglutination (RPLA) and other cytotoxicity assays (Songer, 1996;
Netherwood et al., 1998; Moiler and Ahrens, 1996; Tamiya et al., 1987;
Songer and Meer, 1996; Brynestand et al., 2001). Control measures must
consider two unique attributes of C. perfringens: (i) it grows rapidly at
elevated temperatures, and (ii) formation of spores. Since C. perfringens
will not grow at refrigerated temperatures, the principal cause of virtually
all outbreaks is failure to properly refrigerate the cooked foods. Cooked
foods should be divided into small portions so that refrigeration temperature
is reached within two hours. These foods should be reheated to a minimum
internal temperature of 74C (165F) immediately before serving to destroy
vegetative cells. Cooked meat should be kept above 60C (140F) or below
4C (40F) to restrict the growth of toxic strains. Preventive measures depend
to a large extent on knowledge of proper food preparation and storage,
especially temperature control.

4. CONCLUSIONS
Bacterial food poisoning or intoxication is a major public health problem
world-wide. The majority of food poisoning outbreaks occur as the result of

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poor food hygiene, and improper handling, storage and preservation of

foods and food products. Food handlers and consumers should be educated
to make them familiar with the proper and appropriate methods of food
handling, storage and preservation. Awareness among food handlers and
consumers can be helpful in preventing food poisoning. There is rapid
development of sensitive techniques and methods for the detection of
pathogenic bacteria and their toxins in foods and food products. Researchers
are interested in determining the diversity of microorganisms and their
toxins that are associated with the food-borne illnesses. An effort should be
made to suppress the expression of genes responsible for toxin production
in food-borne pathogens. Equally important is the development of highly
effective and consumer-friendly food preservatives to prevent the growth
of pathogenic and toxigenic microbes in foods and food products.

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