Research Article Phenotypical Identification and Toxinotyping of Clostridium perfringens Isolates from Healthy and Enteric Disease- Affected Chickens Eaftekhar Ahmed Rana , 1 Tanvir Ahmad Nizami, 1 Md Sayedul Islam, 2 Himel Barua, 1 and Md Zohorul Islam 1 1 Chattogram Veterinary and Animal Sciences University, Khulshi, Chattogram•4225, Bangladesh 2 Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur•1706, Bangladesh Correspondence should be addressed to Eaftekhar Ahmed Rana; eaftekhar@cvasu.ac.bd Received 9 November 2022; Revised 28 January 2023; Accepted 31 January 2023; Published 8 February 2023 Academic Editor: Carlos Viegas Copyright © 2023 Eaftekhar Ahmed Rana et al. Tis is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 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. Te 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 identifed 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 classifed 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 identifed 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. Tis is the frst 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. 1. Introduction C. perfringens is an anaerobic, spore forming enteric path• ogen that causes both clinical and subclinical enteric disease in chickens. Te most severe clinical form of enteric disease is necrotic enteritis, which is characterized by a ballooned, friable intestine with necrosis of the intestinal mucosa that is often covered by a tan•to•yellow pseudomembrane [1]. In addition, due to secondary bacterial infection and a rough• ened intestinal mucosal surface, it appears like a Turkish towel [1]. Te clinical form of the disease is associated with a huge economic burden [2], and the subclinical form of the disease signifcantly reduces the growth performance of chickens by causing extensive damage to the gut epithelial layer [3]. Te principal mechanism of disease manifestation by C. perfringens is associated with the release of six major extracellular toxins, which are described as alpha (α), beta (β), epsilon (ε), iota (ι), enterotoxin (cpe), and NetB. Based on the production of the abovementioned toxins, C. perfringens is classifed into seven toxinogenic types, A to G [4, 5]. In the toxinogenic typing scheme, type A and all other types of C. perfringens produce alpha (a) toxin. In Hindawi Veterinary Medicine International Volume 2023, Article ID 2584171, 8 pages https://doi.org/10.1155/2023/2584171