Institute of Microbiology and Molecular Genetics, University of the Punjab, Lahore, 54590, Pakistan.
Department of Animal Sciences, College of Agriculture, University of Sarghoda, Sarghoda, Pakistan; Department of Animal Sciences, CVAS- Jhang Campus, University of Veterinary & Animal Sciences, Lahore, Pakistan.
Anaerobe. 2022 Feb;73:102499. doi: 10.1016/j.anaerobe.2021.102499. Epub 2021 Dec 8.
Clostridium perfringens is a causative agent of enteric infections in animals including poultry by producing twenty different types of toxins. A single strain produces only a subset of these toxins, which form the basis of its classification into seven toxinotypes (A-G). C. perfringens toxinotype A is a widespread cause of necrotic enteritis (NE) in poultry. The current study was conducted to determine the prevalence of different toxins and antimicrobial susceptibility of C. perfringens isolated from Pakistan NE affected poultry.
A total of 134 intestinal samples of the diseased birds were collected postmortem and processed for isolation of C. perfringens using tryptose sulphite cycloserine (TSC) agar supplemented with d-cycloserine. Isolates were confirmed by Gram's staining, biochemical and molecular analyses. Toxinotyping was performed by multiplex PCR. Antimicrobial susceptibility profile of isolates was performed by Kirby Bauer disc diffusion method.
A total of 34 strains of C. perfringens were isolated from 134 samples with prevalence rate of 25.37%. All the isolated strains were toxinotype A, as they were positive for alpha toxin (CPA) and negative for other tested toxins such as beta (CPB), epsilon (ETX), iota (ITX), enterotoxin (CPE), toxin perfringens large (TpeL) and necrotic B-like toxin (NetB). Interestingly, all the isolated strains of C. perfringens were multidrug resistant. The highest resistance was observed against Neomycin, Trimethoprim, Tetracycline and Lincomycin which are routinely used at Pakistan poultry production.
C. perfringens toxinotype A is prevalent in Pakistan poultry. Incidence of C. perfringens with prevalence rate of 25.37% can pose serious threat to Pakistan's poultry industry given that all the isolated strains were multidrug resistant. Our findings highlight the need for new antibiotics and antibiotic alternatives to overcome multidrug resistance.
产气荚膜梭菌是一种动物肠道感染的病原体，包括家禽，通过产生 20 种不同类型的毒素。单一菌株仅产生这些毒素的一部分，这些毒素构成了其分为 7 种毒素型（A-G）的基础。产气荚膜梭菌毒素型 A 是家禽坏死性肠炎（NE）的广泛原因。本研究旨在确定从巴基斯坦 NE 受影响的家禽中分离的产气荚膜梭菌的不同毒素和抗菌药物敏感性。
共采集 134 只患病鸟类的肠道样本，死后用添加 D-环丝氨酸的胰蛋白酶亚硫酸盐环丝氨酸（TSC）琼脂进行产气荚膜梭菌的分离。通过革兰氏染色、生化和分子分析确认分离株。采用多重 PCR 进行毒素定型。采用 Kirby Bauer 纸片扩散法进行分离株的抗菌药物敏感性谱分析。
从 134 个样本中分离出 34 株产气荚膜梭菌，总流行率为 25.37%。所有分离株均为毒素型 A，因为它们对α毒素（CPA）呈阳性，而对其他测试毒素（如β（CPB）、ε（ETX）、ι（ITX）、肠毒素（CPE）、毒素产气荚膜梭菌大（TpeL）和坏死 B 样毒素（NetB）呈阴性。有趣的是，所有分离的产气荚膜梭菌均为多药耐药菌。对新霉素、甲氧苄啶、四环素和林可霉素的耐药性最高，这些药物在巴基斯坦家禽生产中常规使用。
产气荚膜梭菌毒素型 A 在巴基斯坦家禽中流行。产气荚膜梭菌的发病率为 25.37%，可能对巴基斯坦家禽业构成严重威胁，因为所有分离株均为多药耐药菌。我们的研究结果强调需要新的抗生素和抗生素替代品来克服多药耐药性。
