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CLOSTRIDIUM.ppt
- 1. Universities Press © Universities Press (India) Private Limited CLOSTRIDIUM TETANI Hippocrates • 1889 – Kitasato – pure forms reproduced disease – inoculation in animals • Distribution: ubiqutous, soil, intestine of man, animals, hospital dust, cotton, bandages, plaster of Paris, catgut, talc
- 2. Universities Press © Universities Press (India) Private Limited MORPHOLOGY • Drumstick 4–8 x 0.5 ǔm, Gram-positive, motile C.tetani: drumstick appearance
- 3. Universities Press © Universities Press (India) Private Limited CULTURE • Anaerobe • Temperature – 14–43°C, optimum 37°C • RCM • Fresh blood agar • Translucent spreading growth
- 4. Universities Press © Universities Press (India) Private Limited CULTURE • Deep agar shake culture – colonies – spherical with filaments arranged radially • Gelatin stab cultures – fir tree growth • RCM – turbidity with gas formation • Blood agar – α hemolysis, later β hemolysis
- 5. Universities Press © Universities Press (India) Private Limited RESISTANCE • Sensitivity to physical and chemical • Varies – boiling 5–15 minutes, 3 hrs • Iodine 1% • Dry heat 150°C for 1 hr • Resistant to 5% phenol
- 6. Universities Press © Universities Press (India) Private Limited CLASSIFICATION • Ten serological types – I to X • All types produce the same type of toxin which is neutralised by antitoxin produced against any one type
- 7. Universities Press © Universities Press (India) Private Limited TOXINS • Two toxins – hemolysin (tetanolysin) and neurotoxin (tetanospasmin) • Antigenically and pharmacologically distinct
- 8. Universities Press © Universities Press (India) Private Limited TETANOLYSIN • Heat labile • Oxygen labile hemolysin • Produced by C. perfringens, C. novyi and Str. pyogenes
- 9. Universities Press © Universities Press (India) Private Limited TETANOSPASMIN • Responsible – tetanus • Heat labile – inactivated at 65°C in five minutes • Good antigen, specifically neutralised by antitoxin • Toxoided spontaneously, low concentration of formaldehyde
- 10. Universities Press © Universities Press (India) Private Limited TOXIN • Monomer – toxic • Toxin exists • Dimer – non-toxic • Good antigen • Susceptibility : Horses, guinea pigs, mice, goats, rabbits
- 11. Universities Press © Universities Press (India) Private Limited TOXIN Tetanus • Surgical operations • Punctured wounds • Septic abortions • Umblical – sepsis • Ear piercing – unhygienic • Circumcision – conditions
- 12. Universities Press © Universities Press (India) Private Limited FAVOURABLE DISEASE CONDITIONS 1. Anaerobic 2. Presence necrotic 3. Other bacteria 4. Lack of drainage 5. Dose and toxigenicity of strain 6. Site and nature of wound 7. Immune status of host 8. Incubation 6–12 days Mortality rate 80–90 per cent before specific treatment was available Tetanus neonatarum and uterine tetanus 70 per cent
- 13. Universities Press © Universities Press (India) Private Limited TETANUS Pathogenicity • Spore – implanted wound • Multiplies (anaerobic) • Toxin • CNS – via peripheral nerves • Toxin fixed by gangliosides • SP blocks – Syn. inhibition in spinal cord
- 14. Universities Press © Universities Press (India) Private Limited TETANUS • Ascending tetanus – up the spinal cord – opposing hind limbs, trunk, forelimbs • Descending tetanus – IV spasticity of head, neck, limbs • Naturally occurring tetanus
- 15. Universities Press © Universities Press (India) Private Limited NEONATAL TETANUS
- 16. Universities Press © Universities Press (India) Private Limited TETANUS IN HUMAN
- 17. Universities Press © Universities Press (India) Private Limited LABORATORY DIAGNOSIS Confirmation 1. Microscopy BA half plate – polymyxin 1–2 days 2. Culture 80°C for 15 min RCM 3 tubes 80°C for 5 min Unheated 37°C for 4 days
- 18. Universities Press © Universities Press (India) Private Limited SEROTYPING Ten serotypes I–X • VI – Non-flagellate strains Toxins and pathogenicity • Neuro toxin develops – cultures 2–14 at 35°C
- 19. Universities Press © Universities Press (India) Private Limited TOXIGENICITY TEST In vitro • Blood agar plates – one half tetanus antitoxin (1500 units) • C. tetani inoculated on each half of plate • Incubated anaerobically – two days • Toxigenic C. tetani – hemolysis around colonies on half without antitoxin
- 20. Universities Press © Universities Press (India) Private Limited TOXIGENICITY TESTING • In vivo • Control • 500–1500 units • Anti toxin 1 hr before IP • Test • 0.1 ml hind limb • Ascending tetanus
- 21. Universities Press © Universities Press (India) Private Limited PROPHYLAXIS • Tetanus – preventable • Spores – ubiquitous – wound contaminated Prevention 1. Immunisation: Active, Passive, Combined 2. Surgical 3. Antibiotic penicillin, tetracycline
- 22. Universities Press © Universities Press (India) Private Limited CLOSTRIDIUM BOTULINUM BOTULISM • Food poisoning • Botulism – botulum – sausage • Saprophyte – soil, vegetable, animal, hay Morphology • Gram-positive, 5 µ. • Oval, bulging, subterminal spore
- 23. Universities Press © Universities Press (India) Private Limited Culture • Strict anaerobes, 35ºC • Colonies – large, irregular, semi transport • Spore production: Alk, glucose, gelatin 20–25ºC Classification: 8 types A–H • Cl. botulinum – toxin, powerful exotoxin • Toxin – death and lysis • Toxin – synthesized – non-toxic • Pro toxin – converted by enzymes to active toxin, 80–30ºC – 40 min • Toxin inactivated at 100ºC within 10 min CLOSTRIDIUM BOTULINUM BOTULISM
- 24. Universities Press © Universities Press (India) Private Limited TOXIN • Toxin – resists digestion • Absorbed – intestine • Blocks – acetylcholine • Diplopia, dysphasia, responsible for paralysis • Death
- 25. Universities Press © Universities Press (India) Private Limited PATHOGENICITY • Foodborne, wound, infantile • Meat, meat products, canned vegetables, fish • Symptoms – 12–36 hours of ingestion • Vomiting, thirst, constipation, difficulty in swallowing, speaking and breathing • Death – respiratory failure 1–7 days • Wound botulism – toxin – site absorbed
- 26. Universities Press © Universities Press (India) Private Limited INFANTILE BOTULISM • Ingestion of contaminated “honey” • Poor feeding, constipation, loss of head control • Pt excretes – spores and toxin • Laboratory diagnosis – demonstration of spores and bacilli in feces and food • Animal inoculation – guinea pig, mice • Clostridium difficile – large oval subterminal spore – fall antibiotic administration – ampicillin, clindamycin, Tc, lincomycin
- 27. Universities Press © Universities Press (India) Private Limited LABORATORY DIAGNOSIS • Demonstration of toxin in food or feces • C. botulinium – isolated in food or feces • Typing – type-specific toxin
- 28. Universities Press © Universities Press (India) Private Limited C. DIFFICILE • Antibiotic associated colitis • Difficult to isolate • Gram-positive bacillus • Spores – large, oval, subterminal
- 29. Universities Press © Universities Press (India) Private Limited PATHOGENESIS • Acute colitis with bloody diarrhea and pseudomembranous colitis • Common cause of nosocomial diarrhea • Follows the use of broad spectrum antibiotics (clindamycin) to which organism is resistant
- 30. Universities Press © Universities Press (India) Private Limited DIAGNOSIS AND TREATMENT • Diagnosis – demonstration of toxin in feces – Hep2 and human diploid cell cultures – ELISA • Treatment – metranidazole – drug of choice • Vancomycin and bacitracin – useful