COMMON FOODBORNE PATHOGENS

Md. Tarikul Islam

Sub-assistant Manager (Microbiology)
PRAN Quality Control Laboratory, PIP
PRAN-RFL Group
 MICROBIOLOGY
Microbiology: Microbiology is the study of microscopic organisms, such as bacteria,
viruses, archaea, fungi and protozoa.
Purpose of Microbiology:
 Microbiology helps to improve human lives by providing identification-methods and
suggesting treatment methods for all manner of bacterial infections and diseases.
 Microbiologists also help to ensure a clean, healthy food supply and to find purposeful
work in various industries.
Food microbiology: the study of microorganisms causing food spoilage and foodborne
illness. Using microorganisms to produce foods, for example by fermentation.
 FOODBORNE PATHOGEN
Foodborne Pathogens:
Foodborne pathogens are mainly bacteria, viruses, or even parasites that are
present in the food and are the cause of major diseases such as food
poisoning.

Common Foodborne Pathogens:

Some common foodborne pathogens are-
Escherichia coli
Staphylococcus spp.
Salmonella spp.
Bacillus cereus
Listeria monocytogenes
Saccharomyces cerevisiae
Enterobacteriaceae. etc
ESCHERICHIA COLI
 ESCHERICHIA COLI
Important Species of E. coli :
 Shiga toxin-producing E. coli (STEC) relates to serotype O157:H7.
 Uropathogenic E. coli (UPEC) is one of the main causes of urinary tract infections.
 Enterotoxigenic E. coli (ETEC) is the most common cause of traveler's diarrhea.
 EPEC (Gastroenteritis), EIEC (Bacillary dysentery), EHEC (Haemorrhagic colitis).

Sources:
 The reservoir of this pathogen appears to be mainly cattle.
 In addition, other ruminants such as sheep, goats, deer are considered significant reservoirs, while
other mammals (such as pigs, horses, rabbits, dogs, and cats) and birds (such as chickens and
turkeys) have been found infected.
 Primary sources of STEC outbreaks are raw or undercooked ground meat products, raw milk, and
fecal contamination of Foods and vegetables.
 Escherichia coli (E. coli) is a bacterium that is commonly found in the gut of humans and warm-
blooded animals.
 ESCHERICHIA COLI
Foods at risk:
Examples of foods implicated in outbreaks of E. coli O157:H7 include-
 Undercooked hamburgers.
 Unpasteurized fresh-pressed apple cider.
 Yogurt.
 Juice.
 Bakery items.
 Chocolate items.
 Cheese.
 Food items that are made from raw milk.
 Ground, roasted beef, cooked meats.
 Ice cream bars and cakes.
 Raw vegetables, eg., lettuce, sprouts, coleslaw etc.
 ESCHERICHIA COLI
Characteristics:
Escherichia coli is a,
 Gram-negative
 Facultative anaerobic
 Rod-shaped
 Coliform bacterium of the genus Escherichia that is commonly found in the
lower intestine of warm-blooded organisms (endotherms).
 Non- sporulating bacterium.
 β – glucoronidase positive.
 Enteric bacteria.
 ESCHERICHIA COLI
Growth condition:
 Escherichia coli has a optimum growth temperature of 44˚ C - 45.5˚ C with 18
– 24 hours of incubation period. This growth condition is generally used for
recovery of E. coli from foods.
Some STEC can grow in temperatures ranging from 7 °C to 50 °C, with an
optimum temperature of 37 °C.
Some STEC can grow in acidic foods, down to a pH of 4.4, and in foods with a
minimum water activity (aW) of 0.95.
 ESCHERICHIA COLI
Transmission:
Several ways of transmission:
Person to person (via contaminated hands  100 , even as few as 10 cells can
cause disease in highly susceptible populations).
Contact with human or bovine feces.
Water borne.
Cross-contamination from inanimate object.
 ESCHERICHIA COLI

Transmission:
 Beside particulate matter such as ions and molecules, the most common types of contamination are:
 People – Hair, fibre particles from bodies and clothes, also poor hygiene leading to deposition of
microorganisms
 Environment – Dust particles, contaminated air, work surfaces, gases, movement ceilings, walls and
floors
 Materials – Microorganisms on packaging, packaging also creates particles, fibres, dust.
 Equipment – Moving parts shavings drive belts.
 Buildings – Paint flaking, rusty pipe work, poorly maintained surfaces.
 Water – Microorganisms grow in water, equipment not cleaned correctly left in a damp condition,
spills not mopped up properly etc.
 ESCHERICHIA COLI

Illness & Symptoms:

• Most E. coli strains do not cause disease, naturally living in the gut, but virulent strains can
cause gastroenteritis, urinary tract infections, neonatal meningitis, hemorrhagic colitis,
and Crohn's disease.
• Common signs and symptoms include severe abdominal cramps, diarrhea, hemorrhagic colitis,
vomiting, and sometimes fever.
• In rarer cases, virulent strains are also responsible for bowel necrosis (tissue death) and perforation
without progressing to Hemolytic-uremic syndrome (HUS), peritonitis, mastitis, sepsis, and Gram-
negative pneumonia.
• Some strains of E. coli, for example O157:H7, can produce Shiga toxin (classified as
a bioterrorism agent).
• The Shiga toxin causes inflammatory responses in target cells of the gut, leaving behind lesions which
result in the bloody diarrhea that is a symptom of a Shiga toxin-producing E. coli (STEC) infection.
 ESCHERICHIA COLI

Illness & Symptoms:

• Shiga toxin causes premature destruction of the red blood cells, which then clog the body's
filtering system (the kidneys), in some rare cases (usually in children and the elderly)
causing hemolytic-uremic syndrome (HUS), which may lead to kidney failure and even
death.
• Signs of HUS include decreased frequency of urination, lethargy, and paleness of cheeks and
inside the lower eyelids & also may causes acute renal failure, haemolytic anaemia and
thrombocytopenia (low blood platelets).
• In 25% of HUS patients, complications of nervous system occur, which in turn causes strokes.
• In addition, this strain causes the buildup of fluid (since the kidneys do not work), leading
to edema around the lungs and legs and arms. This increase in fluid buildup especially around
the lungs impedes the functioning of the heart, causing an increase in blood pressure.
 ESCHERICHIA COLI

Treatment:
• The mainstay of treatment is the assessment of dehydration and replacement of fluid and electrolytes.
• Administration of antibiotics has been shown to shorten the course of illness and duration of excretion of
enterotoxigenic E. coli (ETEC).
• Though the rate of resistance to commonly used antibiotics is increasing and they are generally not
recommended. Currently, the antibiotics of choice are fluoroquinolones or azithromycin, with an emerging role
for rifaximin.
• Oral rifaximin, a semisynthetic rifamycin derivative, while rifaximin is effective in traveller's diarrhea, it appears
ineffective in patients infected with inflammatory or invasive enteropathogens.
• There are currently no licensed vaccines for ETEC, though several are in various stages of development.
• In different trials, the rCTB-WC cholera vaccine provided high (85–100%) short-term protection.
• An oral ETEC vaccine candidate consisting of rCTB and formalin inactivated E. coli bacteria expressing major CFs has
been shown in clinical trials to be safe, immunogenic, and effective against severe diarrhea in American travelers but
not against ETEC diarrhea in young children in Egypt.
• A modified ETEC vaccine consisting of recombinant E. coli strains over-expressing the major CFs and a more LT-like
hybrid toxoid called LCTBA, are undergoing clinical testing.
 ESCHERICHIA COLI

Control System:
 Maintain Good Hygiene Practice(GHP) / Good Manufacturing Practice (GMP).
 Trained up all personnel related to product handling.
 Preventive measures for E. coli O157:H7 infection are similar to those recommended for other
foodborne diseases. Basic good food hygiene practice, as described in the WHO “Five keys to safer
food”, can prevent the transmission of pathogens responsible for many foodborne diseases, and also
protect against foodborne diseases caused by STEC.
 The five keys to safer food are-
• Keep clean.
• Separate raw and cooked.
• Cook thoroughly.
• Keep food at safe temperatures.
• Use safe water and raw materials.
 ESCHERICHIA COLI
Control System:
 Such recommendations should in all cases be implemented, especially "cook thoroughly" so
that the center of the food reaches at least 70 °C. Make sure to wash fruits and vegetables
carefully, especially if they are eaten raw. If possible, vegetables and fruits should be peeled.
Vulnerable populations (such as small children and the elderly) should avoid the consumption of
raw or undercooked meat products, raw milk, and products made from raw milk.
 A number of STEC infections have been caused by contact with recreational water.
 WHO’s "Five keys to growing safer fruits and vegetables“.
 Other proven prevention methods for E. coli transmission include hand-washing and improved
sanitation and drinking water, as transmission occurs through fecal contamination of food
and water supplies.
 Additionally, thoroughly cooking meat and avoiding consumption of raw, unpasteurized
beverages, such as juices and milk are other proven methods for preventing E. coli. Lastly,
avoid cross-contamination of utensils and work spaces when preparing food.
 ESCHERICHIA COLI

Testing:
• Differential coliform test or Confirmed Escherichia coli count is a test used for the identification
of coliform bacteria from warm-blooded animals based on the bacteria's ability to produce gas
when grown in glucose media at 46 °C (114.8°F).
• E. coli can be tested in Microbiological laboratory on Macconkey Agar media / Eosin
Methylene Blue (EMB) media / Tryptone Bile Glucoronic Agar (TBGA) media at 44±2°C
with an incubation hour of 18-24 hours. We can also use Brilliant Green Bile (BGB) broth and
Gram's Staining procedure.
 ESCHERICHIA COLI

Main Disease Factor:

• Most E. coli strains are harmless, but some can cause serious food poisoning.
• Shiga toxin-producing E. coli (STEC) is a bacterium that can cause severe
foodborne disease.
• STEC produces toxin known as Shiga-toxins. This toxin is the main disease
causing factor of E. coli.
 ESCHERICHIA COLI

Shiga-like toxin (Stx)

Two subunits:
1.Subunit A- enzymatic
2.Subunit B- receptor binding
• toxin inhibits protein synthesis
• Damages vascular endothelial cells in certain organs
• Disrupts the homeostatic properties of these cells
 ESCHERICHIA COLI

Pathogenesis:
1.Establishment of the organism in the gut
2. once established, produce cytotoxins
3. both local and systemic disease mediated
Local intestinal effects:
• toxin internalizes the cells of gut
• blocks cellular protein synthesis
• may lead to apoptosis
• development of bloody diarrhoea

Systemic response:
• the toxins enter the blood stream
• bind to receptors on endothelial cells abundant in kidneys and brain
• Results in HUS
STAPHYLOCOCCUS SPP.
 STAPHYLOCOCCUS SPP.
Important species:
 S. epidermidis is the most important coagulase-negative staphylococcus (CNS) species and is
the major cause of infections associated with prosthetic devices and catheters.
 Multiple antibiotic resistance is increasingly common in S. aureus and S. epidermidis.
 Methicillin resistance is indicative of multiple resistance. Methicillin-resistant S. aureus (MRSA)
causes outbreaks in hospitals and can be epidemic S. aureus (nares) and S. epidermidis (nares,
skin) are common commensals and also have the greatest pathogenic potential.
 S. saprophyticus (skin, occasionally) is also a common cause of urinary tract infection.
 S. haemolyticus, S. simulans, S. cohnii, S. warneri and S. lugdunensis can also cause infections in
man.
 STAPHYLOCOCCUS SPP.

Sources:
 Staphylococcus aureus or “staph” is a type of bacteria found on human skin, in the nose,
armpit, groin, and other areas.
 S. aureus colonizes the nasal passage and axillae.
 S. epidermidis is a common human skin commensal.
 Other species of staphylococci are infrequent human commensals and some are commensals of
other animals.
 STAPHYLOCOCCUS SPP.

Foods at risk:
Foods that are associated with staph food poisoning include:
Meats.
Poultry and egg products.
Salads such as egg, tuna, chicken, potato, and macaroni.
Bakery products such as cream-filled pastries, cream pies, and chocolate eclairs.
Sandwich fillings.
Milk and dairy products.
 STAPHYLOCOCCUS SPP.

Characteristics:
Staphylococcus is a genus of
 Gram-positive bacteria in the family Staphylococcaceae from the order Bacillales.
 Under the microscope, they appear spherical (cocci), and form in grape-like clusters.
 Staphylococcus species are facultative anaerobic organisms (capable of growth both
aerobically and anaerobically).
 Coagulase positive / negative.
 STAPHYLOCOCCUS SPP.

Growth conditions:
 Temperature range: (4 - 46) °C (39-115°F) for growth and toxin production.
 Optimum Temperature: 37° C (98.6° F) pH range: 4.8 - 8.0.

Transmission:
 These bacteria are spread by having direct contact with an infected person, by using a
contaminated object, or by inhaling infected droplets dispersed by sneezing or coughing.
 Skin infections are common, but the bacteria can spread through the bloodstream and infect
distant organs.
 The foot is also very prone to picking up bacteria from the floor.
 STAPHYLOCOCCUS SPP.
Transmission:
 Many healthy people normally have staph on their skin, in their noses, or other body areas. Most
of the time, the germ does not cause an infection or symptoms. This is called being colonized with
staph. These persons are known as carriers. They can spread staph to others. Some people
colonized by staph develop an actual staph infection that makes them sick.
 skin-to-skin contact.
 They can also be spread when you touch something that has the staph germ on it, such as clothing
or a towel. Staph germs can then enter a break in the skin, such as cuts, scratches, or pimples.
 Usually the infection is minor and stays in the skin, but the infection can spread deeper and affect
the blood, bones, or joints.
 Organs such as the lungs, heart, or brain can also be affected.
 Serious cases can be life-threatening.
 STAPHYLOCOCCUS SPP.
Illness:
 A staph infection is caused by a Staphylococcus (or "staph") bacteria.
 The infection often begins with a little cut, which gets infected with bacteria. This can look like
honey-yellow crusting on the skin.
 These staph infections range from a simple boil to antibiotic-resistant infections to flesh-eating
infections. The difference between all these is the strength of the infection, how deep it goes,
how fast it spreads, and how treatable it is with antibiotics. The antibiotic-resistant infections
are more common in North America, because of our over-use of antibiotics.
 One type of staph infection that involves skin is called cellulitis and affects the skin's deeper
layers. It is treatable with antibiotics.
 People who have diabetes or weakened immunity are particularly prone to developing
cellulitis.
 STAPHYLOCOCCUS SPP.
Symptoms:
Symptoms of a staph skin infection can include:
 A painful red lump or bump. This is often a boil or carbuncle (cluster of boils).
 Hot, red and swollen skin. This could an infection called cellulitis.
 Sores, crusts or blisters. This could be impetigo, which often affects the face.
Sore, red eyelids or eyes.
 Staph cellulitis usually begins as a small area of tenderness, swelling, and redness. Sometimes
it begins with an open sore. Other times, there is no obvious break in the skin at all.
 The signs of cellulitis are those of any inflammation - redness, warmth, swelling, and pain.
 If the staph infection spreads, the person may develop a fever, sometimes with chills and
sweats, as well as swelling in the area.
 STAPHYLOCOCCUS SPP.

Treatment:
Antibiotics are used to treat staph infections.
But there's been a gradual change in how well these antibiotics work.
While most staph infections used to be treatable with penicillin, stronger
antibiotics are now used.
 STAPHYLOCOCCUS SPP.

Issues relating to control:

Patients and staff carrying epidemic strains, particularly MRSA, should be
isolated. Patients may be given disinfectant baths or treated with a topical
antibiotic to eradicate carriage of MRSA. Infection control programs are used in
most hospitals.
Follow these steps to avoid a staph infection and prevent it from spreading:
 Keep your hands clean by washing them thoroughly with soap and water. Or use
an alcohol-based hand sanitizer.
 Keep cuts and scrapes clean and covered with bandages until they heal.
 Avoid contact with other people's wounds or bandages.
 Do not share personal items such as towels, clothing, or cosmetics.
 STAPHYLOCOCCUS SPP.

Testing:
Diagnosis is based on performing tests with colonies.
Tests for clumping factor, coagulase, hemolysins and thermostable
deoxyribonuclease are routinely used to identify S. aureus.
Commercial latex agglutination tests are available.
Identification of S. epidermidis is confirmed by commercial bio-typing kits.
 STAPHYLOCOCCUS SPP.
Main disease causing actor:
 α-toxin (septic shock).
 β-toxin.
 δ-toxin.
 γ-toxin and leukocidin.
 Superantigens: enterotoxins and toxic shock syndrome toxin (TSST).
 Epidermolytic (exfoliative) toxin (ET).
 S. aureus can express several different types of protein toxins which are probably responsible for
symptoms during infections.
 Some damage the membranes of erythrocytes, causing hemolysis; but it is unlikely that hemolysis
is relevant in vivo.
 The leukocidin causes membrane damage to leukocytes and is not hemolytic.
SALMONELLA SPP.
 SALMONELLA SPP.

Important species:
 Salmonella enterica – many different serotypes that can be divided into two groups-
i) non-typhoid and
ii) typhoid.
 Salmonellosis, caused by non-typhoid S. enterica is one of the most frequently reported
foodborne disease in the world.
 S. enterica comprises a large number of different serotypes although a few dominate in terms of
causing disease e.g. S. enteritidis, S. typhimurium .
 The typhoid fever causing serotypes are S. typhi and S. paratyphi produce typhoid (enteric)
fever or typhoid-like fever in humans, which is a much more series form of Salmonella
infection.
 SALMONELLA SPP.

Sources:
Non- typhoid Salmonella is ubiquitous in the natural environment i.e. soil,
water. Prevalent in intensive animal/bird farming practices.
Man is the reservoir for S. typhi and S. paratyphi.
Water may become contaminated via human faecal waste.
 SALMONELLA SPP.

Foods at risk:
 Associated in particular with foods of animal origin, such as-
 Poultry meat.
 Eggs.
 Poultry & egg products.
 meat & meat products.
 Cakes & ice cream.
 Milk & dairy products.
 Fish and shrimp.
 Peanut butter.
 Cocoa.
 Chocolate.
 SALMONELLA SPP.
Characteristics and growth conditions:
 Gram negative.
 Rod-shaped.
 Non-spore forming.
 Motile – peritrichous flagella.
 Facultative anaerobic (i.e. can grow in the presence or absence of air).
 Growth temperature (8 – 45)˚ C, Optimum 37˚ C.
 Growth pH 4 – 8 (Optimum 6.5 – 7.5).
 Growth aW - 0.93 .
 Can survive well in foods and on surfaces, for long periods at room temperature or below and for long
periods in animal sewage applied to land (up to 8 months)
 Can survive in dried foods.
 Can survive in broth with 12% salt (22 days at 20˚ C and 55 days at 5˚ C)
 SALMONELLA SPP.

Transmission:
Ingestion of contaminated food.
Contamination of foods with S. typhi and S. paratyphi is usually via food
handlers or fecally contaminated water.
Disseminated via animal and human feces to soil and water.
Salmonella spp. Spread generally by the fecal-oral route and rarely from person-
person.
An estimated 94% salmonellosis is transmitted by contaminated food.
 SALMONELLA SPP.
Illness & Symptoms:
 Non-typhoid salmonella causes Salmonellosis - a gastrointestinal illness which occurs (6 – 48) hours
after ingestion.
 Symptoms include nausea, vomiting, cramps, diarrhea, fever and last (1 – 7) days.
 Can have long term effects (e.g. septicemia may occur) and chronic consequences (e.g. arthritic
symptoms in 3 – 4 weeks).
 All age groups are susceptible, but symptoms are most severe in the elderly, infants, and the immune-
compromised.
 Typhoid occurs 7– 28 days after ingestion of S. typhi cells.
 Symptoms include fever, malaise, anorexia, diarrhea or constipation, delirium.
 Recovery is slow and may take up to 2 months. Hospitalization is often required.
 S. typhi is not a problem in industrialized countries. Those at risk include people living in poor
sanitation conditions and international travelers .
 SALMONELLA SPP.

Doses & Treatment:

 The dose required to cause non-typhoid salmonellosis varies with many factors. While in
general 105 – 106 cells are required to cause illness, doses as low as 4–45 cells have been
reported to cause illness. As with non-typhoid salmonellosis the infectious dose to cause typhoid
fever also appears to be variable.
 Most people with Salmonella recover in 4-7 days and do not need treatment.
 During the illness, the people should drink plenty of fluids to replace the fluids lost by diarrhea.
 A person who has severe diarrhea or is sick for more than a week need to be hospitalized.
 In the hospital he/she will be treated with IV (Intravenous) fluids.
 Antibiotics may be used to treat infants, people over age 65, people with a weak immune system
and those who have severe diarrhea and a high fever and have the bacteria in bloodstream.
 SALMONELLA SPP.

Prevention:
 One of the most heat resistant vegetative cells. D value at 60° C = 2 – 6 minutes Varying heat
resistance- Increased heat resistance following heat shock and also at a lower aW values (e.g. milk
concentrate and chocolate) Freezing usually kills Salmonella, but some foods e.g. meat appear to be
protective of Salmonella. So freezing does not ensure inactivation.
 Inactivated by radiation (D value around 0.5 kGy, up to 0.8.) but effectiveness depends on food type
e.g. D times are higher in drier foods such as desiccated coconut.
 Effect of pH is related to other factors such as acid type and temperature – e.g. inactivation is more
rapid in commercial mayonnaise at 20° C than it is at 4° C.
 Sensitive to disinfectants used in food industry.
 Avoid direct handling of food by infected employees,
 In General- Cook food thoroughly and cool rapidly.
 Keep hot foods above 60° C and cold foods below 4° C.
 Reheat cooked foods to 74° C.
 SALMONELLA SPP.

Testing:
Bismuth Sulphite Agar (BSA).
Xylose lysine deoxycholate Agar (XLD Agar).
Brilliant Green Agar (BGA).
API Kits.
Impedance-conductance techniques and
ELISA kits are also commercially available.
 SALMONELLA SPP.
Main Disease factor:
Salmonella specieses that are Non Typhidal Salmonella (NTS) produces
Cytolethal distending toxin (S-CDT) responsible for typhoid fever.
Salmonella strains may produce a thermolabile enterotoxin that bears a limited
relatedness to cholera toxin both structurally and antigenically. This enterotoxin
causes water secretion in rat ileal loop and is recognized by antibodies against
both cholera toxin and the thermolabile enterotoxin (LT) of enterotoxinogenic E.
coli, but it does not bind in vitro to ganglioside GM1 (the receptor for E. coli LT
and cholera ctx).
Additionally, a cytotoxin that inhibits protein synthesis and is immunologically
distinct from Shiga toxin has been demonstrated. Both of these toxins are
presumed to play a role in the diarrheal symptoms of salmonellosis.
BACILLUS CEREUS
 BACILLUS CEREUS

Important species:
The Bacillus cereus group comprises seven closely related species:
B. cereus sensu stricto (referred to herein as B. cereus),
B. anthracis
B. thuringiensis
B. mycoides
B. pseudomycoides and
B. cytotoxicus
 BACILLUS CEREUS

Sources:
B. cereus is found in
Soil.
Raw plant foods such as- rice, potatoes, peas, beans and
Spices.
 BACILLUS CEREUS
Foods at risk:
Meat products.
Soups.
Vegetables.
Puddings/sauces and milk/milk products.
Fried and cooked rice.
 Pasta.
Pastry and
Noodles.
 BACILLUS CEREUS
Characteristics:
Bacillus cereus is a
 Gram-positive.
 Rod-shaped.
 Facultative anaerobic.
 Motile.
 β – hemolytic.
 Spore forming bacterium.
 BACILLUS CEREUS
Growth condition:
B. cereus grows in a range of (10 – 50) °C, with a temperature optimum
between (30 – 40) °C.
However, individual cold-tolerant strains can also multiply at temperatures of (4
– 6) °C, though with considerably longer generation times.
In dry or acidic foods B. cereus is not able to grow.
 BACILLUS CEREUS

Transmission:
The primary mode of transmission is via the ingestion
of B. cereus contaminated food.
Emetic type of food poisoning has been largely associated with the consumption
of rice and pasta, while the diarrheal type is transmitted mostly by milk
products, vegetables and meat.
 BACILLUS CEREUS
Illness :
 B. cereus is responsible for a minority of foodborne illnesses (2–5%), causing
severe nausea, vomiting, and diarrhea.
 Bacillus foodborne illnesses occur due to survival of the bacterial endospores when infected food
is not, or inadequately cooked.
 Cooking temperatures less than or equal to 100 °C (212 °F) allow some B. cereus spores to
survive. This problem is compounded when food is then improperly refrigerated, allowing the
endospores to germinate. Cooked foods not meant for either immediate consumption or rapid
cooling and refrigeration should be kept at temperatures below 10 °C (50 °F) or above 50 °C
(122 °F). Germination and growth generally occur between 10 °C and 50 °C, though some strains
can grow at low temperatures.
 Bacterial growth results in production of enterotoxins, one of which is highly resistant to heat and
acids (pH levels between 2 and 11); ingestion leads to two types of illness: diarrheal and emetic
(vomiting) syndrome.
 BACILLUS CEREUS
Illness:
 The diarrheal type is associated with a wide range of foods, has an 8-16 hour incubation time, and is associated with
diarrhea and gastrointestinal pain.
 Enterotoxin can be inactivated after heating at 56 °C (133 °F) for 5 minutes, but whether its presence in food causes
the symptom is unclear, since it degrades in stomach enzymes; its subsequent production by
surviving B. cereus spores within the small intestine may be the cause of illness.
 The 'emetic' form is commonly caused by rice cooked for a time and temperature insufficient to kill any spores
present, then improperly refrigerated.
 It can produce a toxin, cereulide, which is not inactivated by later reheating.
 This form leads to nausea and vomiting 1–5 hours after consumption.
 Emetic toxin can withstand 121 °C (250 °F) for 90 minutes.
 B. cereus is also known to cause difficult-to-eradicate chronic skin infections, though less aggressive
than necrotizing fasciitis.
 B. cereus can also cause keratitis.
 BACILLUS CEREUS

Symptoms:
Abdominal pain.
Watery diarrhea and occasionally nausea.
Vomiting and malaise (sometimes followed by diarrhea, due to additional
enterotoxin production).
 BACILLUS CEREUS
Doses:
B. cereus produces beta-lactamases, unlike Bacillus anthracis, and so is resistant
to beta-lactam antibiotics.
It is usually susceptible to treatment with
 Clindamycin.
 Vancomycin.
 Gentamicin.
 Chloramphenicol and
 Erythromycin.
Simultaneous therapy via multiple routes may be required.
 BACILLUS CEREUS
Control system:
• Keep hot foods above 60 °C and cold foods below 4 °C to prevent the formation of
spores.
• Wash hands, utensils, FCSs with hot soapy water after they touch raw meat or poultry, or
before food preparation, and after using the bathroom.
• While B. cereus vegetative cells are killed during normal cooking, spores are more
resistant.
• Viable spores in food can become vegetative cells in the intestines and produce a range of
diarrheal enterotoxins, so elimination of spores is desirable.
• In wet heat, spores require more than 5 minutes at 121 °C (250 °F) at the coldest spot to
be destroyed.
• In dry heat, 120 °C (248 °F) in a sterilizer for 1 hour works for rice, for instance.
 BACILLUS CEREUS

Testing:
• For the isolation and enumeration of B. cereus, there are two standardized methods by International
Organization for Standardization (ISO): ISO 7932:2004 and ISO 21871:2006.
• Because of B. cereus ability to produce lecithinase and its inability to ferment mannitol, there are some
proper selective media for its isolation and identification such as Mannitol-egg Yolk-Polymyxin (MYP)
and Polymyxin-pyruvate-Egg yolk-Mannitol-Bromothymol blue Agar (PEMBA).
• B. cereus colonies on MYP have a violet-red background and are surrounded by a zone of egg-yolk
precipitate.
• Below is a list of differential techniques and results that can help to identify B. cereus from other bacteria
and Bacillus species.
• Anaerobic growth: Positive
• Voges-Proskauer test: Positive
 BACILLUS CEREUS
Testing:
 Acid produced from
 D-glucose: Positive
 L-arabinose: Negative
 D-xylose: Negative
 D-mannitol: Negative

 Starch hydrolysis: Positive

 Nitrate reduction: Positive
 Degradation of tyrosine: Positive
 Growth at
 above 50 °C: Negative

 Use of citrate: Positive

 The Central Public Health Laboratory in the United Kingdom tests for motility, hemolysis, rhizoid growth,
susceptibility to γ-phage, and fermentation of ammonium salt-based glucose but no mannitol, arabinose, or xylose.
 BACILLUS CEREUS
Main disease factors:
 B. cereus causes two different types of food poisoning: the Diarrheal type and the Emetic type.
 The diarrheal type of food poisoning is caused by complex enterotoxins, produced during vegetative
growth of B. cereus in the small intestine, while the Emetic toxin is produced by growing cells in the food.

 The diarrheatic syndromes observed in patients are thought to stem from the three toxins: hemolysin BL
(Hbl), nonhemolytic enterotoxin (Nhe), and cytotoxin K (CytK).
 These genes occur in the taxonomically related B. thuringiensis and B. anthracis, as well.
 The timing of the toxin production was previously thought to be possibly responsible for the two different
courses of disease, but in fact the emetic syndrome is caused by a toxin- cereulide, found only in emetic
strains and is not part of the "standard toolbox" of B. cereus.
 Cereulide is a cyclic polypeptide containing three repeats of four amino acids: D-oxy-Leu—D-Ala—L-
oxy-Val—L-Val similar to valinomycin produced by Streptomyces griseus produced by non-ribosomal
peptide synthesis.
LISTERIA MONOCYTOGENES
 LISTERIA MONOCYTOGENES

Important species:
Listeria monocytogenes is the species of pathogenic bacteria that causes the
infection listeriosis. It is a facultative anaerobic bacterium, capable of surviving in
the presence or absence of oxygen. It can grow and reproduce inside the host's
cells and is one of the most virulent foodborne pathogens: 20 to 30% of foodborne
listeriosis infections in high-risk individuals may be fatal
 LISTERIA MONOCYTOGENES
Source:
 Soil.
 Water.
 Vegetation.
 Feed.
 Industrial plants.
 Farms.
 It can persist along the food continuum.
 It can also be readily isolated from
 Humans.
 domestic animals.
 Raw agricultural and fishery products.
 Food processing environments and homes.
 Feed may be an important source of contamination for animals.
 Domestic and wild animals harbour L. monocytogenes in their intestines and
 The bacterium is also commonly found in food processing environments and in many types of foods
 LISTERIA MONOCYTOGENES

Foods at Risk:
Meat.
Poultry.
Seafood.
Dairy - unpasteurized milk and milk products.
Fruits and vegetables.
 LISTERIA MONOCYTOGENES
Characteristics:
 It is a pathogenic Bacterium.
 Gram-positive.
 Motile.
 Non-spore-forming.
 Highly mobile.
 Rod-type.
 Facultative anaerobic bacterium.
 It tolerates salt and nitrite.
 Grows under low-oxygen conditions and at low refrigeration temperatures.
 LISTERIA MONOCYTOGENES
Growth Condition:
This bacterium grows best between 21.1°C and 37.8°C and slows down
considerably at lower temperature such as those used in refrigeration.
Although the Food Code requires that refrigerated foods be held at 5 °C or
below, the colder the temperature of the food, the greater the impact on limiting
growth of L. monocytogenes.
It is important to get foods cold quickly and to keep them cold.
If low levels of L. monocytogenes are accidentally present in a RTE food item
that supports growth, over time the microorganism can multiply to higher
numbers and pose a significant risk for human health.
 LISTERIA MONOCYTOGENES
Transmission:
Listeria can be spread to people by several different methods. Eating food contaminated with
the bacteria, such as through raw (unpasteurized) milk or contaminated vegetables, is often a
source for cases.
The bacteria may be passed from mother to fetus during pregnancy or directly to the newborn
at the time of birth.
There are some common tools of L. monocytogenes, including the following:
 Food products.
 Environment.
 Equipment.
 Employees.
 customers or vendors.
 LISTERIA MONOCYTOGENES
Illness & Symptoms:
Listeriosis, a most common foodborne disease is caused by L. monocytogenes .
Some common symptoms are include-
Fever.
Chills.
Muscle aches.
If the listeria infection spreads to your nervous system, signs and symptoms can include:
Headache.
Stiff neck.
Confusion or changes in alertness.
Loss of balance.
Convulsions.
 LISTERIA MONOCYTOGENES

Treatment:
For more serious cases of listeriosis, antibiotics are the most common treatment
choice.
Ampicillin can be used alone or in conjunction with another antibiotic (often
gentamicin).
If septicemia or meningitis occur, the individual will be given intravenous
antibiotics and require up to 6 weeks of care and treatment.
 LISTERIA MONOCYTOGENES
Control System:
Essential ways to control L. monocytogenes in retail establishments:
 Prevent cross-contamination.
 Practice proper sanitation and
 Control time and temperature.
A clean, dry environment is of utmost importance in controlling Listeria.

Testing:
Listeria Agar base selective media is used for isolation of this bacteria.
 LISTERIA MONOCYTOGENES

Main disease factors:

Listeriolysin O (LLO) is a toxin produced by Listeria monocytogenes, an opportunistic bacterial
pathogen responsible for the disease listeriosis.
This disease starts with the ingestion of contaminated foods and mainly affects
immunocompromised individuals, newborns, and pregnant women.
Other L. monocytogenes toxins have been described to date, including phospholipases (PlcA,
PlcB) that contribute to the escape from the endocytic and secondary vacuoles, the
thiazole/oxazole-modified toxin Listeriolysin S (LLS) that behaves as a bacteriocin favoring
intestinal colonization, or the toxin/antitoxin MazEF involved in growth and survival under
stress.
In addition, several L. monocytogenes secreted or surface-associated proteins play major roles in
virulence.
SACCHAROMYCES CEREVISIAE
 SACCHAROMYCES CEREVISIAE

Important Species:
 Saccharomyces cerevisiae is a species of Yeast (single-celled fungus microorganisms).
 The species has been instrumental in wine making, baking, and brewing since ancient times.

Sources:
 Saccharomyces when translated means “sugar fungus”. That is what this yeast uses for food.
 They are found in the wild growing on the skins of grapes and other sugar containing fruits.
 They may also found in air while they are accompanied with vectors/ air particles.
 SACCHAROMYCES CEREVISIAE
Food at risk:
 Fresh fruits and vegetables.
 Nuts.
 Beans.
 Cereals.
 Meat and dairy products.
 Flour.
 Pasta.
 bakery products.
 jams and preserves.
 Soft drinks and alcoholic beverages.
 Confectionary.
 Canned food.
 SACCHAROMYCES CEREVISIAE
Characteristics:
 Colonies of Saccharomyces grow rapidly and mature in three days.
 They are flat, smooth, moist, glistening or dull, and cream in color.
 The inability to use nitrate and ability to ferment various carbohydrates are typical characteristics of Saccharomyces.
 Generally, they have a diameter of 2-8 μm and length of 3-25 μm.
 Blastoconidia (cell buds) are observed.
 They are unicellular, globose and ellipsoid to elongate in shape.
 Multilateral (multipolar) budding is typical.
 Pseudohyphae, if present are rudimentary. Hyphae are absent.
 Saccharomyces produces ascospores, especially when grown on V-8 medium, acetate ascospor agar, or Gorodkowa
medium.
 These ascospores are globose and located in asci., each ascus contains 1-4 ascospores. Asci do not rupture at
maturity.
 Ascospores are stained with Kinyoun stain and ascospore stain. When stained with Gram stain, ascospores appear
Gram-negative, while vegetative cells appear Gram-positive.
 SACCHAROMYCES CEREVISIAE

Growth Condition:
The best growth conditions for Saccharomyces cerevisiae are: 30 °C, pH 5.0, a
higher concentration yeast than 2×10-2 gm/L and a glucose concentration of 22
gm/L gave the best doubling time of 2 hours and 57 minutes.
The pH is calibrated with 2 buffers pH 4 and 7.
 SACCHAROMYCES CEREVISIAE

Transmission:
 Saccharomyces cerevisiae was found in the skin, oral cavity, oropharinx, duodenal mucosa,
digestive tract, and vagina of healthy humans (one review found it to be reported for 6% of
samples from human intestine).
 Some specialists consider S. cerevisiae to be a part of the normal microbiota of the
gastrointestinal tract, the respiratory tract, and the vagina of humans, while others believe that the
species cannot be called a true commensal, because it originates in food.
 Presence of S. cerevisiae in the human digestive system may be rather transient, for example,
experiments show that in the case of oral administration to healthy individuals it is eliminated
from the intestine within 5 days after the end of administration.
 SACCHAROMYCES CEREVISIAE

Illness & Symptoms:

Many cases of fungemia due to S. cerevisiae (well known as “baker's yeast” or
“brewer's yeast”)
S. cerevisiae have been reported in patients with chronic disease, cancer, and
immunosuppression.
Endocarditis.
Pneumonia.
Peritonitis.
urinary tract infections.
Skin infections and esophagitis have been described.
It is important to consider infections due to S. cerevisiae in appropriate clinical settings.
 SACCHAROMYCES CEREVISIAE

Treatment:
S. cerevisiae includes administration of antifungal agent and removal of infected
foreign bodies, especially CVC.
The antifungal agent of choice for treatment of Saccharomyces species has not
been finally established, but amphotericin B and fluconazole seems to be
preferable.
 SACCHAROMYCES CEREVISIAE

Control System:
 Maintaining of GMP (Good manufacturing Practice) or GHP (Good hygiene practice).
 WHO recommended 5 keys to grow safer fruits and vegetables-
 Practice good personal hygiene.
• Protect fields from animal fecal contamination.
• Use treated fecal waste.
• Evaluate and manage risks from irrigation water.
• Keep harvest and storage equipment clean and dry.
 Maintaining of controlled air circulating system.
 Neat & Clean environment.
 SACCHAROMYCES CEREVISIAE

Testing:
S. cerevisiae is tested accordance with ISO 21527 – 1 & 2:2008.
Dichloran Rose Bengal Chloramphenicol agar (DRBC Agar) is used and
incubated at 25±3 ˚C for 120 hours.
 SACCHAROMYCES CEREVISIAE

Main disease factor:

S. cerevisiae strains redirect the low levels of the nitrogen sources inside the
phagosome to increase amino acids biosynthesis.
These strains also increase the oxidative stress response by activating the Yap1p
transcription factor regulon to counteract the lethal effects of reactive oxygen
species (ROS) generated by the macrophage.
De novo dNTP biosynthesis is activated to allow DNA repair machinery to
counteract ROS-induced mutations.
ENTEROBACTERIACEAE
 ENTEROBACTERIACEAE
Important Species:
 Salmonella spp.
 Escherichia coli.
 Klebsiella spp.
 Shigella spp.
 Enterobacter spp.
 Citobacter spp.
 Yersinia spp.
 Erwinia spp.
 Pectobacterium spp.
 Proteus spp.
 ENTEROBACTERIACEAE

Sources:
Normal member of gut microiota in human and animals (Gastrointestinal tract).
Naturally found in
 Water.
 Soil.
 Surfaces and
 Plants.
Some are parasites on different animals and plants.
 ENTEROBACTERIACEAE
Foods at risk:
 Eggs.
 Vegetables.
 Rice.
 Beef.
 Chicken.
 Potatoes.
 Pies.
 Street foods.
 Water.
 Raw milk.
 Cheese.
 Dairy products.
 Time and temperature abused foods.
 ENTEROBACTERIACEAE
Characteristics:
Bacilli/rod shaped.
Maximum are Motile (have flagella).
Non-spore forming.
Facultative anaerobes.
Sugar fermenting and produce lactic acid.
Lack of cytochrome-C oxidase.
Reduce Nitrate to Nitrite.
Catalase positive.
Oxidase negative.
 ENTEROBACTERIACEAE

Growth Condition:
Growth temperature ranges 5 to 40 ˚C with an optimum temperature of 37 ˚C.
Can grow with or without presence of Oxygen.
 ENTEROBACTERIACEAE
Transmission:
 Contact with wounds.
 Stools.
 Clothes.
 Direct ingestion.
 Flees.
 Pests.
 Personnel.
 Uncontrolled/unhygienic handling of products.
 Contaminated water.
 ENTEROBACTERIACEAE

Illness:
Wound infection.
Urinary tract infection (UTI).
Gastroenteritis.
Meningitis.
Pneumonia (can lead to a possible lung abscess, pura-pneumonic pleural effusion)
Septicemia.
Respiratory tract infection (RTI).
All infection from Enterobacter spp. May be complicated by sepsis & septic
shock.
 ENTEROBACTERIACEAE

Sign & symptoms:

Shortness of breath (pneumonia).
Pain with urination (UTI).
Pain & swelling of skin (skin infection).
Abdominal cramp (Liver infection).
Stiff neck and reduced consciousness (meningitis).
Fever, chills and fatigue (any local infection).
 ENTEROBACTERIACEAE
Treatment:
Carbapenem, β – lactams, β – lactamase inhibitors, Fluroquinolones, Aminoglycosides and
Sulphamethoxazole / Trimethoprime.
1st & 2nd generation cephalosporin are generally not effective against Enterobacter infections.
3rd & 4th generation cephalosporin are generally not recommended against Enterobacter
infections as they are specific for some specieses.
Carbapenem (Meropenem & Imipenem) shown to be the most potent treatment for multi-
drug resistant Enterobacter infection.
Possible treatment for Carbapenem-resistant Enterobacter (CRE) include- ploymyxins,
tigecycline, fosfomycin and Carbapenem ( used in a double regimen).
 ENTEROBACTERIACEAE

Control system:
GMP / GHP.
Food items store at elevated temperature.
Controlled handling of raw ingredients.
 ENTEROBACTERIACEAE

Testing:
Agar plating and biochemical tests accordance with ISO 21528-1&2:2017 for
isolation and identification of Enterobacteriaceae from foods and food
processing related samples.
Solid media containing Bile salt and glucose (such as Violet Red Bile Agar)
 ENTEROBACTERIACEAE

Main disease factor:

 Extended spectrum β- lactamase (ESBL) leads to-
 Liver abscess.
 Poly-microbial infection.
 Bacteremia.
 Pulmonary infection.
 Recurrence.
 After orthotropic liver transplantation (OLT) causes Enterobacteriaceae bacteremia..
 Enterotoxins- α hemolytic, thiol-activated pore-forming cytotoxins, similar to shiga like toxin 11,
which required. treatment with 2-mereaptotheonal.
 ESBL also lead to subsequent EPE-BSI (Blood stream infection).
THANK YOU
……………