Subject: Microbiology Topic: Bacteria Assoc. w/ GIT Infx. 3 (food poisoning) Lecturer: Dr. Solis Date of Lecture: Nov.

08, 2011 Transcriptionist: kekie Editor: I <3 demsey Pages: 5

Outline Food Poisoning A. Staphylococcal Food Poisoning B. Clostridial Food Poisoning 1. Clostridium spp. a. Clostridium perfringes b. Clostridium botulinium C. Bacillus cereus Food Poisoning 1. Bacillus spp. a. Bacillus cereus FOOD POISONING ingestion of pathogenic bacteria as well as their toxins important factor: no. of patient who consumed the food who exhibited the same signs and symptoms causative agents: 1. Staphylococcus 2. Clostridium 3. Bacillus

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A. STAPHYLOCOCCAL FOOD POISONING caused by Staphylococcus aureus the MOST COMMON food-borne illness more of an intoxication rather than an infection because it is CAUSED BY TOXINS rather than the effect of the bacteria on the patient. Epidemiology: o Found in skin, nasopharynx, GIT, urogenital tract o Can survive on dry surface for long periods o MOT: ingestion of preformed enterotoxin in food (ex. contaminated potato salad, mayonnaise, processed meat[hotdogs, ham]) o Diseases: Food poisoning TSS SSSS Wound infections Pneumonia Endocarditis Bacteremia Septic arthritis Morphology: o Gram (+) cocci o Grape-like clusters, singly or in pairs o 0.5-1.5 micrometer o facultative anaerobes o non-motile Antigenic Structures / Virulence Factors o capsule o peptidoglycan o techoic acid o Protein A

Cytotoxin Exfoliative toxin TSS toxin-1 Coagulase Catalase Hyaluronidase Lipase Nuclease Penicillinase Enterotoxins 8 serologically distinct enterotoxins (A-E, G-I) Enterotoxin A is the most commonly associated with food poisoning Enteroxin B- pseudomembranous colitis It takes 25 mcg of enterotoxin B to produce symptoms(diarrhea, vomiting, etc)- I know its confusing, its not in our book either.. she said that the cause of food poisoning is enterotoxin A but she mentioned enteroxin B causing the symptoms above. According to medscape, the cause of food poisoning is enterotoxin B, but according to Doc Solis its Enterotoxin A. o.O 0 HEAT STABLE (can withstand heat at 100 C for 30 minutes) Note: the bacteria will be killed but its toxin will still be active despite heating. Resistant to hydrolysis Mechanisms not well understood Induce non-specific activation of T-cells and cytokine release

Pathogenesis: Infected host sneezes on food Incubation for about 6 hours Organism produces enterotoxin Food eaten by susceptible host After 1-8 hours: diarrhea, abdominal pain, nausea, vomiting SELF LIMITING (patient gets well after about 24 hours)

Laboratory Diagnosis: o G/S and C/S NOT usually done because it is self limiting o Lab work-ups are done in cases of mass intoxication and only done for documentation o Specimen: contaminated food

SY 2011-2012

Cultural/growth characteristics: Sheep blood agar (added with 7.5% sodium chloride to kill other microorganisms) - Golden, large, smooth, round, raised and glistening colonies - Produces beta hemolysis Biochemical tests (+) catalase (+) coagulase (+) heat stable nuclease (+) alkaline phosphatase (+) mannitol fermentation

Treatment o Supportive- ex. Give medication for abd pain o Fluid replacement- for fluid loss from vomiting and diarrhea o Antibiotic is NOT indicated Prevention and Control o proper hand washing o covering of exposed skin surfaces o refrigeration of food o

Antigenic Structures/ Virulence Factors Major toxins (lethal toxins) - toxin o Aka lecithinase o lyses RBC, platelets, leukocytes, endothelial cells. o Increases vascular permeability o Tissue destruction o C. perfringens type A produce large amounts of alpha toxin - toxins- necrotic lesions (necrotizing enterocolitis) - toxins- increase vascular permeability of GI wall - toxin- necrotic activity, increase vascular permeability Enterotoxin HEAT LABILE protein released during bacterial lysis when releasing spores - transformation from vegetative to spore formation will cause the release of enterotoxin - alters membrane permeability of the ileum and jejunum by inhibiting fluid absorption Pathogenesis: Ingestion of meat, fish, poultry Sporulation after 8-24 hours Enterotoxin produced in the GIT, stimulated by alkalinity of GIT(remember it alters the ileum and jejenum, which have an alkaline envi) Abdominal cramps, watery diarrhea Self-limiting

B. CLOSTRIDIAL FOOD POISONING caused by: 1. Clostridium perfringens 2. Clostridium botulinum other species: o C. defficile (cause pseudomembranous colitis) o C. tetani (cause tetanus) o C. septicum o C. novyi o C. sordelli o C. tertium o C. histolyticum o C. barati o C. butricum Clostridium spp. o Gram (+) bacilli, anaerobic o Spore-forming: produce oval, subterminal spores, except for C. tetani (terminal) o Motile or non-motile rods

1. Clostridium perfringens o 5 types: A,B, C, D, E o Type A: related to food poisoning o Type C: enteritis neroticans o Epidemiology: Found in intestinal tract of humans and animals MOT: Soil and water contaminated with feces Diseases: soft tissue infections, necrotizing enterocolitis, food poisoning, septicemia Morphology: Gram (+) bacillus Large, rectangular (box-car shaped) Capsule (+/-) spores non-motile strictly anaerobe

Laboratory Diagnosis usually not done (since it is self limiting) specimen: o food ( presence of >105 organisms per gram of food is confirmatory) o feces (presence of > 106 bacteria per gram of feces is confirmatory) cultural/growth characteristics - Strictly anaerobeic - Blood agar (27 C) - Thioglycolate, cooked meat broth - Colonies on BAP: - Large raised colonies with entire edges - (+) double zone of hemolysis - inner complete zone of hemolysis - outer incomplete/partial zone of hemolysis Biochemical test: - catalase negative Treatment supportive oral rehydration salts (ORS) may be given antibiotic therapy is unnecessary (self-limiting)

Prevention and Control refrigeration of food reheating of food to destroy the heat labile enterotoxins handwashing

2. Clostridium botulinum o Epidemiology: easily found in soil and water samples MOT: associated with consumption of homecanned/ vacuum-packed foods and preserved fish diseases: infant botulism wound botulism food bourne botulism the usual history is ingestion of fruit juice with contaminated honey o Morphology: gram (+) bacilli spore forming strict anaerobe o Antigenic structure/virulence factors Spore formation - Spores of C. botulinum are highly resistant to heat, withstanding 100C for at least 3-5 hours - Heat resistance is diminished at acid pH or high salt concentration Botulinum toxin - Seven types: A, B, C, D, E, F, G Types A,B,E,F causes human disease Types C, B are associated with animal diseases Types A, B poison by canned foods Type E poison by preserved fish - Toxins are liberated during growth of C. botulinum and during autolysis of the bacteria - Among the MOST highly toxic substance known - Blocks neurotransmission at peripheral cholinergic synapses by preventing release of neurotransmitter acetylcholine Binary toxin - Disrupts vascular permeability o Pathogenesis Ingestion of food with pre-formed neurotoxin Neurotoxin absorbed by mucosa of stomach and upper GIT Blood stream Absorbed by peripheral neurons at myoneuronal junction Botulinum blocks SNAP/SNARE which is responsible for the release of transmitter from vesicle Paralysis of pharyngeal, ocular, respiratory muscles, abdominal pain, constipation Death due to respiratory paralysis

Laboratory diagnosis Usually no signs and symptoms Specimen: contaminated food, feces, blood (serum) Cultural/growth characteristics - Enriched anaerobic media Biochemical properties: - (+) lipase production - (+) hydrolyze gelatin - (+) ferment glucose - (+) digest milk proteins other tests - direct toxin activity, - Mouse bioassay Definite test but not usually available in regular labs - Radio immunoassay Treatment ventilatory support (because of respiratory paralysis) gastric lavage - remove organism and toxin - a procedure used to empty gastric contents - put a tube intranasally until it reaches the stomach, then evacuate gastric contents antibiotic therapy: penicillin, metronidazole BOTULINUM ANTITOXINS: give 2 vials once suspected - one vial IM and one vial IV Prevention and Control refrigeration of food heat food (20 minutes at 80C)

C. BACILLARY FOOD POISONING 50 species: o Bacillus anthracis o Bacillus cereus (food poisoning) o Bacillus myoides o Bacillus thurigensis Bacillus spp. o Large Gram (+) bacilli o Strict anaerobes o Seen in soil, water, dust samples o Can produce endospores o Diverse metabolic type and nutritional requirements, composition, and structure of vegetative cell Bacillus cereus o Epidemiology Ubiquitous organism MOT: ingestion for RICE, meat, vegetables Diseases: gastroenteritis, ocular infections, intravenous catheter-related sepsis, systemic infections, food poisoning

Morphology Gram (+) bacilli Spore-forming (subterminal spores) Motile Facultative anaerobe Produces toxins that are more of an intoxication rather than food bourne infection Virulence factors 2 forms of enterotoxins: Emetic Diarrheal EMETIC FORM Heat stable (unknown mechanism) Rice Occasionally pasta Vomiting, nausea, abdominal cramps <6hours (mean=2) 8-10 hours ( mean =9) DIARRHEAL FORM Heat labile (adenylate cyclase cyclic ADMP system) Meat and vegetables

Laboratory diagnosis Usually not done; c/s only done for documentation Specimen: implicated food (rice, meat, vegetables) stool (in case of cluster of patients) gram stain morphology: Gram (+) bacilli (+) motility Cultural/growth characteristics gray round colonies, swarming patter resistant to penicillin Treatment Symptomatic treatment Fluids- diarrhea Anti-spasmodic- cramps Anti-emetic- emetic Some doctors give antibiotics Prevention and control Immediate consumption of food after cooking Proper refrigeration

Enterotoxin

Food

Symptoms

Diarrhea, nausea, abdominal cramps

Incubation Duration

> 6 hours (mean=9) 20-36 hours (mean = 24)

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other virulence factors spore production cereolysin A hemolysin Phospholipase C A lecithenase Lyses RBC and platelets Pathogenesis Emetic form: Contaminated rice Cook rice: most bacilli killed but spore survive

Cooked rice refrigerated: spores germinate, bacilli multiply and heat stable enterotoxin is released Rice is reheated: enterotoxin not destroyed Ingestion of enterotoxin 6 hours incubation Vomiting, nausea, abdominal cramps Diarrheal form: Contaminated meat, vegetables More than 6 hours incubation, organisms multiply Production of heat labile enterotoxin Diarrhea, nausea, abdominal cramps