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Abstract This study was done on 728 intestine and liver samples collected from 84 commercial broiler chicken farms aged from 1-11 weeks-old located at Behera, Kafr El-Sheikh and EI-Gharbia govemorates. Anaerobic bacteriological examination revealed isolation of 430 Clostridial isolates that identified into;C.perfringens(54.9%),C.sporogenese(18.8%),C.sordellii(8.6%),C.tertium(15.3) ,C.bifermentans(1.9%) and C.chaeuvoei (0.5%). Out of the 236 isolated C.perfringens; 61 isolates were toxigenic that subtyped into types A (83.6%), B (1.6%), C (1.6%), and D (13.1 %). C.perfringens also recovered from waterers, feeders and litter surrounding the birds with a percentage of 27.3%, 50% and 22.7%; respectively (5 isolates from them were toxigenic and of type D). The mortality rate reached to 100% in 5 days-old broiler chicks, intramuscularly inoculated with broth culture of C.perfringens type A, D and mixed inoculum of type A and D. The multiplex-PCR assay could amplify c, ~ and € toxins of C.perfringens in an arrangement identical to types A, B, C and D. These results suggested that the multiplex-PCR as a rapid, accurate and humanitarian assay could replace the seroneutralization methods with mice and guinea pigs in detection and typing of toxigenic C.perfringens. In vitro sensitivity test on the toxigenic types of C.perfringens to some antibiotics indicated that, all types were highly sensitive to rifamycin, ceftiofure, ampicillin, amoxycillin, penicillin-G and a combination of amoxycillin and kitamycin, while, they were resistant to streptomycin, oxytetracyclin and erythromycin. |