Vibrio the organism and laboratory diagnosis

Vibrio cholera with other vibrio species are described in thispresentation along with their biochemical properties and laboratory diagnosis, A short slide on halophilic vibrios is also added

1. VIBRIO

2. INTRODUCTION Gram Negative Oxidase positive Short, rigid, curved rods ACTIVELY MOTILE - DARTING Polar flagellum Vibrio cholerae

3. VIBRIO CHOLERAE MORPHOLOGY Non sporing Non capsulated 1.5 * 0.2 Darting motility

4. GRAM STAIN OF PURE CULTURE OF Vibrio cholerae

5. CULTURE Strongly aerobic ALKALINE medium, 8.2 pH Ordinary Media Nutrient Agar – Moist, Translucent, round discs, 1-2mm, bluish tinge Mac Conkeys Agar – Colourless, at first and become red on prolonged incubation

6. BLOOD AGAR – Classical – no hemolysis El Tor – Hemolysis PEPTONE WATER – Surface pelicle

7. l tellurite

8. SPECIAL MEDIA TRANSPORT ( High pH) Venkatraman Ramakrishnan VR Medium Cary Blair Medium

9. If no transport medium – blot stool on thick blotting paper ENRICHMENT MEDIUM Alkaline Peptone water Monsur’s taurocholate Tellurite Peptone Water

10. PLATING SPECIAL MEDIA Alkaline Bile Salt Agar Modified NA – bile salt added, pH 8.2 Monsurs Gelatin Taurocholate Trypticase Tellurite Agar (GTTA )

11. Thiosulphate Citrate Bile Salt Sucrose (TCBS) Most widely used pH 8.6 Contents Sodium Thiosulphate Sodium Citrate Bile Salts Sucrose Bromothymol Blue Yeast Extract Peptone Sodium Chloride Ferric Citrate Water

12. BIOCHEMICAL REACTIONS Catalase -- + Oxidase -- + Glucose -- + , No gas Lactose -- - Mannitol -- + Sucrose -- + Maltose -- + Mannose -- + Indole -- + CHOLERA RED REACTION NO 3 redn -- + Urease -- - Gelatin -- + Lysine -- +

13. BIOCHEMICAL REACTIONS MR -- - VP -- - Ornithine -- + Arginine -- - Sheep RBC hemolysis -- -

14. ANTIGENIC STRUCTURE Somatic O Ag Flagellar H Ag – Common Gardner And Venkatraman Classification O – Major antigen – 139 Minor antigens – 3 ( A,B,C)

15. VIBRIO GROUP A Biochemically similar having common H Antigen ( Vibrio cholerae ) GROUP B Biochemically and antigenically heterogenous Serogroup 01 Non 01 (02- 138) Classical El Tor Ogawa Hikojima Inaba

16. TOXINS Endotoxin Enterotoxin Exotoxin , heat labile Cholera toxin / Choleragen 01 and 0139 produce toxin 2 fractions – A ( 1) and B ( 5) ETEC , but > Potent

17. Mechanism of Action PROBLEMS

18. NON 01 VIBRIOS 02 – 138 – NAG vibrio Not differentiate by culture and biochem V. Cholerae 0139 Madras1992 India & Bangladesh Like El Tor Capsulated

19. DISEASE CHOLERA

20. Cholera hospital in Dhaka, showing typical " cholera beds" EPIDEMIC PANDEMICS – 6 Classical vibrio – Severe Seasonal March – May

21. LABORATORY DIAGNOSIS SPECIMENS Rice watery stools Rectal swab COLLECTION AND TRANSPORT B4 antibiotics Sterile containers Immediately transported to Lab Transport Medium - ?

22. DIRECT MICROSCOPY Darting motility CULTURE Selective Media - BSA, TCBS, Monsur’s GTTA Non Selective Media – Blood Agar Mac Conkey Agar Enrichment Media – APW / Monsurs liquid Medium

23. COLONY MORPHOLOGY AND STAINING Mac conkey – Pale Monsurs Medium – black with halo TCBS – yellow BSA – translucent Gram negative – comma BIOCHEMICAL REACTIONS AGGLUTINATION TEST

24. Killed Parentral Vaccine ( Ogawa & Inaba ) Killed Oral vaccine – B subunit whole cell Live Oral Vaccine – Recombinant DNA

25. Preventive inoculation against cholera in 1966

26. WHO SHOULD GET VACCINATED

27. HALOPHILIC VIBRIOS NEED NaCl Habitat – Sea Water V. parahaemolyticus , V. vulnificus Marine food Kanagawa Phenomenon AEROMONAS/ PLESIOMONAS Gram Neg , motile, oxidase positive A. hydrophila , P. shigelloides

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