Clostridium perfringens (Veillon and Zuber) Hauduroy et al.
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2. Using a single tube of #1053 broth (5 to 6 ml), withdraw approximately 0.5 to 1.0 ml with a Pasteur or 1.0 ml pipette. Rehydrate the entire pellet.
3. Aseptically transfer this aliquot back into the broth. Additional tubes may be inoculated with 0.5 ml each from the suspension. Also, 0.1 ml may be inoculated onto a slant. Streak several blood plates to check for colonial morphology and purity.
4. Incubate tubes under an anaerobic atmosphere at 45oC for 24-48 hours. Incubate one agar plate anaerobically for colony formation and one aerobically at 37oC for aerobic contamination check.
Anaerobic conditions for transfer may be obtained by either of the following:
·Use of an anaerobic gas chamber, or
·Placement of test tubes under a gassing cannula system connected to anaerobic gas.
Anaerobic conditions for incubation may be obtained by any of the following:
·Loose screw caps on test tubes in anaerobic chamber,
·Loose screw caps on test tubes in an activated anaerobic gas pack jar, or
·Use of sterile butyl rubber stoppers on test tubes so that an anaerobic gas headspace is retained.
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Meer RR, Songer JG. Multiplex polymerase chain reaction assay for genotyping Clostridium perfringens. Am. J. Vet. Res. 58(7): 702-705, 1997. PubMed: 9215442
Garmory HS, et al. Occurrence of Clostridium perfringens beta2-toxin amongst animals, determined using genotyping and subtyping PCR assays. Epidemiol. Infect. 124(1): 61-67, 2000. PubMed: 10722131