Phenotypical Identification and Toxinotyping of <i>Clostridium perfringens</i> Isolates from Healthy and Enteric Disease‐Affected Chickens
Eaftekhar Ahmed Rana, Tanvir Ahmad Nizami, Md. Sayedul Islam, Himel Barua, Md Zohorul Islam
Abstract
Clostridium perfringens is a ubiquitous spore‐forming anaerobic pathogen that is frequently associated with enteric disease in chickens. Moreover, enterotoxin‐producing C. perfringens has high zoonotic potential as well as serious public health concerns due to the emanation of food‐borne intoxication. The present study was designed to isolate, identify, and toxinotype C. perfringens from both healthy and cases of necrotic or ulcerative enteritis chickens. A total of 110 samples were collected from July 2019 to February 2021. Among the samples, 38 (34.5%, 95% CI: 26.39–43.83) were positive for C. perfringens and were obtained from broiler 21 (33.3%, 95% CI: 22.91–45.67), Sonali 9 (34.6%, 95% CI: 19.31–53.88), and layer 8 (38%, 95% CI: 20.68–59.20). C. perfringens was highly prevalent (35.7%, 95% CI: 25.48–47.44) in enteritis chickens compared with healthy ones. In multiplex PCR toxinotyping, 34 (89.4%) isolates were identified as C. perfringens type A by the presence of the alpha toxin gene ( cpa ). Moreover, in addition to the cpa gene, 3 (14.3%, 95% CI: 4.14–35.48) broiler and 1 (11.1%, 95% CI: 0.01–45.67) Sonali isolates harbored the enterotoxin gene ( cpe ) and were classified as type F. However, none of the isolates carried genes encoding beta ( cpb ), epsilon ( etx ), iota ( iap ), or beta‐2 ( cpb2 ) toxins. Multivariable logistic regression analysis identified the following variables such as; “previously used litter materials” (OR 21.77, 95% CI 2.22–212.66, p ≤ 0.008); intestinal lesions, “presence of ulceration” (OR 30.01, 95% CI 3.02–297.91, p ≤ 0.004); “ballooned with gas” (OR 24.74, 95% CI 4.34–140.86, p ≤ 0.001) and “use of probiotics” (OR 5.24, 95% CI 0.74–36.75, p ≤ 0.095) act as risk factors for C. perfringens colonization in chicken gut. This is the first study of molecular toxinotyping of C. perfringens from healthy and enteric‐diseased chickens in Bangladesh, which might have a potential food‐borne zoonotic impact on human health.