养殖虾塘常见耐药菌的分离鉴定与耐药基因检测  

Antibiotic Resistant Bacterial Isolation and Identification from Shrimp Ponds and Their Antibiotic Resistance Genes Detection

作　　者：叶繁 冯时欢 吴佳佳 戴志远 YE Fan;FENG Shihuan;WU Jiajia;DAI Zhiyuan(College of Seafood,Zhejiang Gongshang University,Hangzhou 310012,China;College of Life Science,China Jiliang University,Hangzhou 310018,China;State Key Laboratory of Aquatic Products Processing of Zhejiang Province,Hangzhou 310012,China)

机构地区：[1]浙江工商大学海洋食品研究院,浙江杭州310012 [2]中国计量大学生命科学学院,浙江杭州310018 [3]浙江省水产品加工技术研究联合重点实验室,浙江杭州310012

出　　处：《生态环境学报》2019年第9期1843-1849,共7页Ecology and Environmental Sciences

基　　金：浙江省食品药品监管系统科技计划项目（2018001）;浙江省水产品加工技术研究联合重点实验室开放基金项目（2017003)

摘　　要：了解水产养殖区中抗生素耐药菌的分布及抗生素耐药基因的污染现状,有助于科学、合理地使用抗生素。采用诺氟沙星抗性平板和红霉素抗性平板统计虾塘泥样和水样中耐药微生物数量,从中筛选得到60株耐药菌株。基于16S rDNA序列分析完成部分菌株的鉴定,同时,对耐药菌株携带的喹诺酮类(诺氟沙星)抗性基因gyrA及大环内酯类(红霉素)抗性基因ermB进行了扩增及测序分析。结果表明,虾塘水样细菌在诺氟沙星抗性平板中均未生长,虾塘泥样细菌对诺氟沙星的耐药率分别为0.36%、0.82%;1号虾塘泥样和水样中细菌对红霉素的耐药率分别为23.33%、18.50%,2号虾塘泥样和水样中细菌对红霉素的耐药率分别为20.00%、12.50%。耐药菌16S rDNA鉴定结果如下:南极适冷菌(Rheinheimera sp.)7株、不动杆菌(Acinetobacter sp.)6株、微杆菌(Microbacterium sp.)3株、厦门希瓦氏菌(Shewanella xiamenensis)2株、嗜水气单胞菌(Aeromonas hydrophila)2株、豚鼠气单胞菌(Aeromonas caviae)1株。1号虾塘gyrA检出率为43.33%,ermB检出率为30.00%;2号虾塘gyrA检出率为10.00%,ermB检出率为70.00%。耐药基因序列比对结果显示,所测菌株序列与数据库中相应耐药基因序列相似性均≥97%。In order to understand the pollution status of antibiotic resistance genes in shrimp ponds and guide the rational usage of antibiotics in aquaculture process. 60 bacterial isolates were picked up from the sediment and water samples collected from two shrimp ponds using 2216E agar containing antibiotics (norfloxacin, erythromycin). Partial of the isolates were identified based on 16S rDNA gene sequencing. Meanwhile, quinolone resistance genes (gyrA) and macrolide resistance genes (ermB) were amplified and sequenced. The bacteria in water sample from No.1 and No.2 shrimp ponds showed no growth on 2216E agar containing norfloxacin, while norfloxacin resistance rates of bacteria from sediment of No.1 and No.2 shrimp pond were 0.36% and 0.82%, respectively. The erythromycin resistance rates of bacteria from No.1 shrimp pond’s sediment and water samples were 23.33% and 18.50%, respectively, while the resistant rates to erythromycin of No.2 shrimp pond’s sediment samples and water samples were 20.00% and 12.50%. The results of 16S rDNA sequencing indicated that 7 Rheinheimerasp., 6 Acinetobacter sp., 3 Microbacterium sp., 2 Shewanella xiamenensis, 2 Aeromona shydrophila and 1 Aeromonas caviae were identified. The detection rates of gyrA and ermB in No.1 shrimp pond were 43.33% and 30.00%, while those of No.2 shrimp pond were 10.00% and 70.00%. The sequencing result of the antibiotic resistant gene amplicons obtained high similarity compared those in the genebank (similarity≥97%).

关 键 词：抗生素耐药菌 诺氟沙星 红霉素 耐药基因 

分 类 号：X172[环境科学与工程—环境科学]