BIO 425 Industrial Microbiology

Lecture 8 Pathogenic microorganisms in food industry

Content outline
Foodborne intoxications Exotoxins Serotypes of E.coli

Foodborne intoxications
occurs when a person gets sick by eating food: that has been contaminated with an unwanted m/o or toxin. Often called food poisoning.

Most common symptoms of foodborne illnesses

Stomach cramps Nausea Vomiting Diarrhoea Fever

Causes of food poisoning

inadequate control of temperature during cooking, cooling and storage, poor personnel hygiene, cross-contamination of raw and processed products, inadequate monitoring of processes.

Members of normal intestinal flora

Bacteroides spp. (B.fragilis), G(-) non-spore former rods-dominant m/o produce volatile & non-volatile fatty acids, acetic, succinic, lactic etc. Bifidobacterium spp. (B.bifidum) - G(+) non-spore former rods produce acetic & lactic acids

Members of normal intestinal flora-2

Clostridium spp.(C.innocuum)-G(+) spore forming rods. Enterococcus spp.(E. faecalis) G(+) cocci, aerotolerant. Eubacterium spp.-G(+) non-spore forming rods, produce butyric, formic & acetic acids. Fusobacterium spp. G(-) non-spore forming rods, produce N-butyric acid.

Members of normal intestinal flora-3

Peptostreptococcus spp., G(+) cocci, degrade peptone & amino acids. Ruminococcus spp., G(+) cocci, produce acetic, succinic and lactic acids, ethanol, CO2 & H2 from CH4.

Other genera of organisms present:

A minimum of 400 species, includes: Lactobacillus spp. Proteus spp. Klebsiella spp. E.coli spp.

Pathogenic strains involved in major outbreaks of food poisoning:

E.coli G(-), Catalase(+), Oxidase(-), facultatively anaerobic short rods. members of the family Enterobacteriaceae.

More about E.coli

Most isolates ferment lactose, majority of E.coli serotypes: not pathogenic are part of the normal intestinal flora, about 106 organisms / g. used as an indicator: as one of the coliforms to indicate faecal pollution of water, raw ingredients and foods.

Non-pathogenic E.coli
colonize the infant gastrointestinal tract within a few hours after birth, its presence suppresses the growth of harmful bacteria; important for synthesizing appreciable amounts of B-vitamins.

Toxic strains of E.coli

Three major types: Exotoxins, Enterotoxins, Endotoxins.

Exotoxins of E.coli
Same as the bacterial protein toxins, also called secreted toxins; exert their biological activity (often lethal) in minute doses. extracellular, excretory, heat-labile, antigenic.

Exotoxins of E.coli
Exotoxins which result in extremely watery diarrhoea: act as: directly on gut tissue, causing biochemical and /or structural lesions which lead to diarrhoea. causing specific action (e.g. net fluid loss) in the gut. elevate cAMP levels, causing ion-flux changes and excess fluid secretion.

Enterotoxins of E.coli
Two types: Cytotonic enterotoxins, Cytotoxic enterotoxins.

Cytotonic enterotoxins
E.g. Cholera toxin Induce net fluid secretion: by interfering with biochemical regulatory mechanisms; without causing overt histological damage.

Cytotoxic enterotoxins
Induce actual damage to intestinal cells as a necessary prelude to onset of net fluid secretion; can be protein or lipopolysaccharide; invasive, kill target cells. Mode of action: intracellular or by formation of pores within cells. Result: inflammatory diarrhoea which often contains blood and leucocytes.

Endotoxins of E.coli
Heat-stable, cell associated, complex lipopolysaccharide (LPS) structure; Lipid A: Part of LPS which is responsible for toxicity. cause toxic shock, inflammation and fever.

Cytolethal distending toxins (CLDT)

Affect host cell cycle regulation, found in a variety of unrelated organisms. Campylobacter jejuni E.coli Shigella dysenteriae Haemophilus ducreyi Actinobacillus actinomycetemcomitans

Cytolethal distending toxins (CLDT)

Blocks cells in G2 phase: inhibits the dephosphorylation of the protein kinase cdc2; and this prevents entering into mitosis. Belongs to a variety of exotoxin family.

Cytolethal distending toxins (CLDT)

Exotoxins: If they attack a specific organ, they are named as; Neurotoxins, Leukotoxin; Hepatotoxin; Cardiotoxin;

Specific examples for exotoxins:

Cholera toxin by Vibrio cholerae Shiga toxin by Shigella spp. cause bacterial dysentery. Shiga toxin: E.coli O 157: H7 called also verotoxin, as it affects mammalian Vero cell culture line.

Groups of exotoxins
Act at the cell membrane, type I, E.coli stable toxin, B.cereus emetic toxin. attack the membrane: type II; Listeriolysin O (Listeria monocytogenes) Perfringolysin O (Clostridium perfringens) Cereolysin O (B.cereus) E.coli heat labile toxin & shiga toxin.

Groups of exotoxins
Penetrate the membrane to act inside the cell: type III. e.g. Botulinum toxins type A-F & tetanus toxin, cholera. directly transported from the bacterium into the target eucaryotic cell by type III secretion. e.g. Toxins of Salmonella, Shigella & Yersinia.

Campylobacter jejuni & C.coli

Colonises the distal ileum and colon of the human intestinal tract; after colonizing the mucus; and adhering to intestinal surfaces.

Campylobacter jejuni & C.coli

The organisms perturb the normal absorptive capacity of the intestine; by damaging epithelial cell function; by cell invasion & production of heat labile enterotoxin & cytotoxin, and cytoskeletal altering toxin.

Campylobacter jejuni & C.coli

can easily cross-contaminate processed food. A contaminated piece of raw meat can leave 10000 Campylobacter cells per cm2 on a work surface. G(-), thin rod, microaerophilic. 42-43 C-optimum T.

Campylobacter jejuni & C.coli

Symptoms: Flu-like illness, abdominal pain, fever, diarrhea.

Campylobacter jejuni & C.coli

Potential for spreading the infection: Water, milk, meat, poultry.

Salmonella
Member of Enterobacteriaceae, G(-), facultatively anaerobic, non-spore former, Motile with peritrichious flagella-flagella inserted at many places around the cell surface.

Salmonella
Over 2324 strains are defined. Symptoms of food poisoning: Diarrhoea, nausea, abdominal pain mild fever and chills sometimes vomiting and headache.

Major species of Salmonella

Salmonella typhimurium Salmonella enteritidis Salmonella paratyphii Salmonella typhii

Major sources of Salmonella

Raw meat, Poultry, Milk & dairy products, Fish, Shrimp, Sauces, Salad dressings.

Pathogenic E.coli
Enterotoxigenic E.coli (ETEC): common cause of travellers diarrhoea. watery diarrhoea, rice-water like: low grade fever, colonizes the proximal small intestine.

Enteropathogenic E.coli (EPEC)

Cause watery diarrhoea of infants, no blood, colonizes the microvilli over the entire intestine.

Enterohaemorrhagic E.coli (EHEC)

cause bloody diarrhoea, cause haemorrhagic colitis, cause haemolytic uremic syndrome(HUS), cause thrombotic thrombocytopenic purpurea (TTP).

Enterohaemorrhagic E.coli (EHEC)

Includes the verotoxigenic E.coli (VTEC): also known as shiga-toxin producing E.coli (or STEC) serotypes 0157, 026 and 0111.

Enteroaggregative E.coli (EAggEC)

Cause persistent watery diarrhoea; especially in children, lasting more than 14 days. produce a heat labile toxin, antigenically related to haemolysin, but not haemolytic and a plasmid-encoded heat stable toxin (EAST1), unrelated to the heat stable enterotoxin of ETEC.

Enteroaggregative E.coli (EAggEC)

Adhere to the intestinal mucosa and elaborate the enterotoxins and cytotoxins, which result in secretory diarrhoea and mucosal damage.

Enteroinvasive E.coli (EIEC)

Cause fever and diarrhoea, containing mucus and blood. colonizes in the colon and carries a 120-140 mDa plasmid(Invasiveness plasmid), carrying all the genes necessary for virulence.

Most important one from E.coli

Enterohaemorrhagic E.coli (EHEC); E.coli O157: H7 with common outbreaks; O157 and H7 refers to the serotyping of the strains O and H antigens, respectively.

E.coli O157: H7
Differs from the majority of E.coli strains that it does not grow, or grows poorly at 44 C. and does not ferment sorbitol or produce glucuronidase. cause severe damage to the lining of the intestine by verotoxin (VT) & shigatoxin (ST).

Thrombotic thrombocytopenic purpurea(TTP)

Blood platelets surround internal organs leading to: kidney damage, and central nervous system damage.

Thrombotic thrombocytopenic purpurea(TTP)

The most vulnerable of the populationchildren and the elderly develop haemorrhagic colitis (HC); which may lead to haemolytic uraemic syndrome (HUS).

Haemorrhagic colitis (HC)

Less severe form of E.coli O157:H7 infection than HUS. associated with: sudden onset of severe crampy abdominal pains. non-bloody (watery) diarrhoea, vomiting.

Haemolytic Uremic Syndrome(HUS)

Patient suffers from bloody diarrhoea; haemolytic anemia; kidney disorder; renal failure (somehow requires dialysis & blood transfusions).

Haemolytic Uremic Syndrome(HUS)

Myocarditis, other cardiovascular complications that may result in death. central nervous system involvement, hypertension.

Reservoir of E.coli
Raw or undercooked meat products, raw milk, fresh pressed apple juice or cider, yogurt, cheese, salad vegetables, transmission through direct contact between people and faecal contamination of water.

LITERATURE CITED
Forsythe, S.J. The Microbiology of Safe Food, Blackwell Science, Cambridge, 2000. Wood, B. J. B., Microbiology of Fermented Foods, Vol.1 and 2, Blackie Academic and Professional, London, Second edition, 1998.