CLOSTRIDIUM DIFFICILE
CLOSTRIDIUM DIFFICILE: Clostridium difficile is recently accepted widely as the most important cause of severe antibiotic associated diarrhoea. Cross contamination is the most likely explanation for spread of this organism in hospitalized infants. The organism may possibly spread among adults who are at risk of developing antibiotic associated diarrhoea or colitis. Susceptible individual when exposed to this bacteria, it proliferates, release toxin and may produce diarrhoea colitis etc.
For the isolation of this organism stools are cultured immediately on brain heart infusion agar (BHIA) or cycloserine-cefoxitin agar (CCA) media. These plates are incubated at 37°C in the atmosphere of 10% CO2, 10% H2 and 80% N2 for atleast 48 hours in anaerobic incubator. After incubation, plates are examined under long wave (360nm) ultraviolet light for light green fluorescence the characteristic of C. difficile colonies. At least 3 colonies from each plate are subcultured into Robertson Cooked Meat medium. The culture may be identified after 3 days of incubation by use of volatile fatty acid profile of carbohydrate metabolism generated by gas liquid chromatography and biochemical reaction pattern. The cellular morphology of Clostridium difficile is characteristic with apparent bipolar sporing noted in Gram stain of 3 day old culture giving dumb bell appearance.
However isolated Clostridum difficile are tested for production of toxin in vitro using tissue cultures and for production of toxin in vivo by inoculation into guinea pig and clindamycin treated hamsters. This toxin is a necrotising glycoprotein with molecular weight of 50,000. Animal work has shown that certain antibiotic, like vancomycin are useful and deserves trial on human being. Trial of antitoxin may also prove useful as Clostridium difficile toxin can be neutralised by several clostridial ‘toxins’ including that of Clostridium sordelli.
Tags: antepartum, bilirubin, cordblood, jaundice, oedema, pericardial effusions, peritoneal, Pleural
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