Serological and Molecular Typing of Clostriduim Perfringens and Its Toxins Recovered from Weaned Rabbit's Flocks in Egypt

This study was carried out for serological and molecular typing of Clostridium perfringens (C. perfringens) and its toxins recovered from apparently healthy, diseased and dead weaned rabbits, as well as their feed and water. Identified 42 C. perfringens organisms representing 35 from rabbits and 7 from feed and water were subjected for determination of toxigenicity by intravenous inoculation (I/V) in Swiss mice. Toxigenic C. perfringens organisms were typed serologically using Nagler's test, dermonecrotic reaction in albino Guinea pigs and toxin antitoxin serum neutralization test (SNT) in Swiss mice. For confirmation of serological results, detection of alpha gene of C. perfringens was done using conventional polymerase chain reaction (PCR), followed by using of multiplex PCR to detect the toxin's types. Distribution of different C. perfringens types of surveyed rabbit's farms at different Egyptian governorate was carried out. In addition, the in-vitro antibiotic sensitivity of different C. perfringens types against different antimicrobial agents was performed. The results showed that 34 (97.14%) of C. perfringens recovered from rabbits were toxigenic, while 1.0 (2.86%) was none. Four out of 6 (66.66%) C. perfringens isolates from feed was toxigenic and 2 (33.33%) isolates was non-toxigenic. Only one (100%) C. perfringens isolate from water was toxigenic. Identical proving results were obtained using serological typing tests. Single and mixed types of toxigenic C. perfringens constituted 17 out of 35 (48.57%) for each, whereas only one type was none (2.86%). Single C. perfringens was representing 8 (22.85%) type A, 3 (8.57%) type B, 4 (11.43%) type D and 2 (5.71%) type E, whereas mixed types were 11 (31.42%) types (A and D), 2 (5.71%) types (A and E) and 4 (11.42%) types (B and D). Four C. perfringens isolates from feed was 3 type A (50%) and 1.0 type D (16.6%). The only C. perfringens isolate from water was type A (100%). Conventional PCR proved detection of alpha gene of C. perfringens, whereas multiplex PCR proved that C. perfringens type (A) was positive for alpha toxin at 324 base pair (bp), type (B) was positive for alpha toxin at (324 bp), beta toxin at (196 bp) and epsilon toxin at (655 bp), type (D) was positive for alpha toxin at (324 bp) and epsilon toxin at (655 bp) while type (E) was positive for alpha toxin at (324 bp) and iota toxin at (446 bp). The result of distribution of different C. perfringens types at different Egyptian governorate was recorded. In vitro antibiotic sensitivity test of single or mixed types of identified C. perfringens revealed sensitivity to amoxicillin/ clavulanic acid and ampicillin and resistance to colistine, erythromycin and lincomycine. (Khelfa D. E. -D. G., Wafaa A. Abd El-Ghany and Heba M. Salem Serological and Molecular Typing of Clostriduim Perfringens and Its Toxins Recovered from Weaned Rabbit's Flocks in Egypt. Life Sci J 2012;9(4):2263-2271) (ISSN:1097-8135). http://www.lifesciencesite.com. 336