الفهرس 
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Abstract
Clostridium perfringens is a widespread spore-forming, Gram-positive, anaerobic, nonmotile rod causing a wide range of diseases. It is recognized as an enteritic bacterial pathogen in humans, poultry, other farm animals and wildlife worldwide. Species belonging to Clostridium can be subtyped into five toxinotypes based on the production of four major toxins namely α, β, epsilon and iota toxin. In addition to the so-called major toxins, there are at least 13 minor toxins or enzymes produced by some strains of C. perfringens, which may play a role in pathogenicity. These compounds include beta2, netB, delta, theta, kappa, lambda, mu, nu, gamma, eta, neuraminidase, urease and enterotoxin.
A total of 137diseased broiler chicken samples including 52 liver samples and 85 intestinal samples were examined in this study.These samples were collected from different farms and small holders at El-Behera and Kafr El-Sheikh Governorates for anaerobic isolation and identification of C. perfringens.The recovered isolates were biochemically identified by ordinary biochemical tests.
The incidence of C. perfringens isolated from diseased broiler chickens from intestine and liver was 71.8 % (61 out of 85) and 48.1% (25 out of 52) respectively with total percentage 62.8 % .
The prevalence of Clostridium perfringens isolated from diseased broilers in Kafr-El Sheikh Governorate was 57 out of 89 with percentage 64% and in El-Behera Governorate was 29 out of 48 with percentage 60.4%.
The prevalence of Clostridium perfringens isolated from diseased broilers in different ages was 40.7% at 10-20 days old, 77% at 21-30 days old and 50% over 30 days old.
This study indicated higher prevalence of Clostridium perfringens through individuals (housekeeping) than in farms with percentage 72.7% and 60.9% respectively.
Multiplex PCR was used for genotyping of C. perfringens by using four sets of primers specific for toxin-producing genes of C. perfringens (alpha, beta, epsilon and iota). Forty C. perfringens isolates were examined by multiplex PCR for genotyping of C. perfringens. The results revealed that all of the examined field isolates were given alpha toxin (402bp). Thirty nine (97.5%) of the examined isolates revealed alpha toxin gene (402bp) only and were identified as type A. And only 1 (2.5%) of the examined isolates revealed alpha toxin gene (402bp) and Iota toxin gene (317bp) and was identified as C. perfringens type E. On the other hand, all C. perfringens isolates gave no band with beta and Epsilon toxin primers exhibiting the absence of C. perfringens types B, C and D. This result indicates that C. perfringens type”A” is the most prevalent type in NE affected broilers.
The same 40 C. perfringens isolates were examined by monoplex PCR for presence of Net.B and T.pel genes and the result revealed no bands with Net.B and T.pel toxin genes primers as the 40 C. perfringens isolates were examined by monoplex PCR for the netB toxin gene using specific primer and the result revealed no bands at 560bp.and also The same 40 isolates were examined by uniplex PCR for the Tpel toxin gene using specific primer and the result revealed no bands at 466bp.