牛奶中产气荚膜梭菌的分离鉴定和带菌量测定  

Identification and Ddetermination of the Amount of Clostridium perfringens in Milk

作　　者：潘奕帆 刘变芳[1] PAN Yi-fan;LIU Bian-fang(College of Food Science and Engineering,Northwest A&F University,Yangling,Shaanxi,712100,China)

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100

出　　处：《动物医学进展》2020年第7期53-57,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31972043)。

摘　　要：调查陕西关中地区牛奶中产气荚膜梭菌的污染情况及其优势血清型和携带的毒素基因。通过对3个奶牛场采集的86份牛奶样品进行细菌分离纯化,应用多重PCR鉴定分离菌株的血清型,进行cpb2、cpe毒素基因检测,并对场3的乳样按照GB 4789.13-2012规定的方法进行产气荚膜梭菌带菌量的测定。结果显示,个体乳样产气荚膜梭菌的分离率为11.3%(7/62),且全部为A型菌,57.1%(4/7)的分离菌株携带atyp.cpb2基因,但所有分离菌株均未检测到cons.cpb2和cpe基因。而混合乳样(24份)中未分离出目的菌。场3的28份乳样中3份产气荚膜梭菌阳性乳的带菌量为1 CFU/mL^15.3 CFU/mL。初步了解了陕西关中地区生鲜牛奶中产气荚膜梭菌的污染情况、血清型及其携带的毒素基因,为关中地区牛奶中该菌污染的防控提供科学依据,为该菌引起食物中毒的流行病学研究提供参考资料。In order to investigate the serotypes,toxin gene and contamination of Clostridium perfringens in milk in Guanzhong district of Shaanxi Province.Total 86 milk samples were collected from three dairy farms from Guanzhong district of Shaanxi province,C.perfringens were isolated and purified and the isolates were then serotyped by the multiplex PCR and genotyped by cpb2 and cpe gene detection.Further,the amount of C.perfringens in milk samples from farm 3 were detected according to the method specified in GB 4789.13-2012.The results showed that the prevelance of C.perfringens was 11.3%(7/62)from individual milk samples,all isolates were identified as type A.The atyp.cpb2 gene was detected from 57.1%(4/7)of the isolates,but no cons.cpb2 and cpe has been detected from all of the isolates.However no C.perfringens was isolated from 24 mixed milk samples.The amount of C.perfringens in 3 positive samples out of 28 samples were 1 to 15.3 CFU/mL.In this study,the contamination of C.perfringens in milk from Guanzhong district of Shaanxi was initially understood,the dominant serotype is type A,and it provides a scientific basis for the prevention and control of the contamination of C.perfringens in the milk,hence,provides reference information for the epidemiological study of food poisoning caused by C.perfringens.

关 键 词：牛奶 产气荚膜梭菌 分离鉴定 聚合酶链反应 毒素基因 

分 类 号：S823.91[农业科学—畜牧学] TS252.2[农业科学—畜牧兽医]