Sugar Fermentation Test Essay

1100 Words 5 Pages
1.7.3.4. Hydrogen sulphid test (MacFaddine, 1980):
Tubes of triple sugar iron agar media were inoculated by suspected culture through stabbing into the butt and the slant then covered with sterile melted paraffin wax and incubated anaerobically for 48 h at 37°C, appearance of blackening in the butt indicates H2S production.
1.7.3.5. Sugar fermentation test (Willis, 1977):
Suspected culture was inoculated to 1% peptone water tubes containing 2% bromocresol purple and 1% quantities of the following sugar: mannose, glucose, lactose, maltose, mannitol, sucrose and xylose were added. The tubes covered with sterile layer of paraffin wax and incubated anaerobically at 37°C for 7 days.
Table (A): Koneman et al. (1992) scheme
The suspected purified
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Neutralization test in Swiss mice (Smith and Holdeman, 1968):
Swiss mice with an average weight of about 15-20 g were kept under observation for 2 weeks before they were inoculated. The test was performed by adding 0.1 ml of specific antisera to 0.3 ml of the centrifuged supernatant cooked meat culture of each type of clostridium perfringens (A, B, C, D and E).
In case of clostridium perfringens type D the supernatant fluid was treated by 0.1% trypsin for 45 minutes at 37°C. The supernatant fluid of the cooked meat culture without adding any antiserum was used as control. The toxin and anti-toxin mixture was kept for 45 minutes at 37°C before its injection as 0.5 mi intraperitoneally into two mice. Animals were kept under observation for 1-3 days.
1.7.5. Demonstration of C. perfringens toxin by dermonecrotic test (Sterne and Batty, 1975):
1.7.5.1. Preparation of toxin and their treatment (Bullen,
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of trypsinized 48 hrs. supernatant of each culture were intradermally injected and the neutralized one was injected into the left side in the same manner and arrangement. The injected Guinea pigs were kept under observation for 48-72hrs. to demonstrate any dermal reaction. The results were interperitated by the degree of dermonecrotic reaction and its neutralization according to Sterne and Batty (1975) as

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