陕西省、甘肃省和宁夏回族自治区羊源D型产气荚膜梭菌特征分析  

Characterization of Clostridium perfringens type D from sheep sources in Shaanxi,Gansu and Ningxia

作　　者：袁媛 吴克 张雯 崔生玲 杨奇 杨丹娇 奎军 杨文欢[5] 冯航 方明锦 田欣 王兴龙[1,6,7] 王娟[1,6,7] 杨增岐[1,6,7] YUAN Yuan;WU Ke;ZHANG Wen;CUI Shenglin;YANG Qi;YANG Danjiao;KUI Jun;YANG Wenhuan;FENG Hang;FANG Mingjin;TIAN Xin;WANG Xinglong;WANG Juan;YANG Zengqi(College of Veterinary Medicine,Northwest A&F University,Yangling,Shaanxi 712100,China;Ningxia Hui Autonomous Region Veterinary Drug and Feed Supervision Institute,Yinchuan 750011,China;Animal Husbandry Science Institute of Ganzi Tibetan Autonomous Prefecture,Kangding,Sichuan 626000,China;Animal Husbandry and Veterinary Station of Huangzhong District of Xining City,Xining 811600,China;Shaanxi Center for Animal Disease Control and Prevention,Xi'an 710016,China;Key Laboratory of Prevention and Control of Major Ruminant Diseases,Ministry of Agriculture and Rural Affairs(Western Region),Northwest A&F University,Yangling,Shaanxi 712100,China;Engineering Research Center of Novel Animal Efficient Vaccines of the Ministry of Education,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]宁夏回族自治区兽药饲料监察所,宁夏银川750011 [3]甘孜藏族自治州畜牧业科学研究所,四川康定626000 [4]西宁市湟中区畜牧兽医站,青海西宁811600 [5]陕西省动物疫病预防控制中心,陕西西安710016 [6]西北农林科技大学农业农村部反刍动物重大疫病防控重点实验室(西部),陕西杨凌712100 [7]西北农林科技大学动物高效新型疫苗教育部工程研究中心,陕西杨凌712100

出　　处：《中国兽医学报》2024年第5期987-993,999,共8页Chinese Journal of Veterinary Science

基　　金：陕西畜禽育种“两链融合重点专项”资助项目(2022GD-TSLD-46);四川省科技计划资助项目(2023YFN0044);宁夏回族自治区重点研发计划资助项目(2021BEF02041)。

摘　　要：对2020-2022年分离自我国陕西省、甘肃省和宁夏回族自治区的30株D型产气荚膜梭菌进行毒素基因鉴定、耐药性检测、消毒剂敏感性测试和菌株遗传特征分析。毒素基因鉴定显示分离菌株均携带毒素基因cpa和etx,不携带其他分型相关毒素基因。分离菌株对磺胺异噁唑和复方新诺明抗性较强,耐药率分别为30.00%和63.33%;对青霉素、利奈唑胺、克林霉素、多西环素、恩诺沙星、氟苯尼考敏感,MIC值集中于1 mg/L以下。分离菌株经推荐使用浓度的新洁尔灭、乙醇、84消毒液和浓戊二醛处理10 min后,均可被完全杀灭。经多位点序列分析(MLST),30株D型产气荚膜梭菌中共发现7种不同的管家基因组合;同场区和同省(区)来源的菌株间有相同的管家基因组合和更近的进化关系,并且陕西省和甘肃省的部分菌株间也有相同管家基因组合,揭示了D型产气荚膜梭菌在羊场内部和不同省份之间的传播。综上,本研究在一定程度上在基因水平揭示了D型产气荚膜梭菌的地域性流行和传播,提示在生产中应加强对消毒剂和抗生素的合理使用,以减少D型产气荚膜梭菌的传播。This study conducted toxin gene identification,drug resistance detection,disinfectant susceptibility testing and genetic characterization analysis of 30 strains of Clostridium perfringen type D isolated from four sheep farms in Shaanxi,Gansu and Ningxia of China during the period of 2020 to 2022.Toxin gene identification showed that all strains carried the toxin genes cpaand etx,and did not carry other typing-related toxin genes.The drug resistance detection showed that the isolates were more resistant to sulfisoxazole and cotrimoxazole,with the resistance rate of 30.00%and 63.33%,respectively;they were sensitive to penicillin,linezolid,clindamycin,doxycycline,enrofloxacin and florfenicol,with the MIC values concentrated below 1mg/L.The isolates were completely killed after treatment with concentrations of neosporin,ethanol,84disinfectant and concentrated glutaraldehyde for 10mins.After multilocus sequence typing(MLST)analysis,seven different housekeeping gene combinations were identified among the 30strains of C.perfringens type D;the same housekeeping gene combinations and closer evolutionary relationships were often found between strains originating from the same field area and from the same province,and the same housekeeping gene combinations were found between some strains from Shaanxi and Gansu Provinces,which revealed the spread of C.perfringens type D within the sheep farms and between different provinces.In conclusion,this study revealed the geographical prevalence and transmission of C.perfringens type D to a certain extent at the genetic level,suggesting that the rational use of disinfectants and antibiotics should be strengthened in production to reduce the transmission and pathogenesis of C.perfringens type D.

关 键 词：D型产气荚膜梭菌 耐药性 消毒剂抗性 遗传特征 

分 类 号：S852.61[农业科学—基础兽医学] R535[农业科学—兽医学]