水产用微生物制剂源细菌的分离鉴定及耐药性分析  

Isolation,identification and antibiotic resistance analysis of bacteria derived from aquatic microbial agents

作　　者：王红莉 刘丽娟 张德锋[2] 尹纪元 石存斌[2] 任燕[2] 夏苏东 王庆[2] WANG Hong-li;LIU Li-juan;ZHANG De-feng;YIN Ji-yuan;SHI Cun-bin;REN Yan;XIA Su-dong;WANG Qing(Aquaculture Institute,Tianjin Agricultural University,Tianjin 300384,China;Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences/Key Laboratory of Fishery Drug Development,Ministry of Agriculture and Rural Affairs,Guangzhou 510380,China)

机构地区：[1]天津农学院水产学院,天津300384 [2]中国水产科学研究院珠江水产研究所,农业农村部渔用药物创制重点实验室,广州510380

出　　处：《淡水渔业》2023年第4期12-25,共14页Freshwater Fisheries

基　　金：国家大宗淡水鱼产业技术体系专项资金(CARS-45);2021年乡村振兴战略专项资金;广东省现代农业产业技术体系创新团队建设专项资金(2019KJ150);广州市科技计划项目(201904020004)。

摘　　要：微生物制剂因其具有无毒害、无污染、低成本的优点在水产领域迅速发展,成为最具潜力的抗生素的替代产品。本研究通过对市场在售水产微生物制剂中的细菌进行分离培养与鉴定,以及抗生素药敏实验、耐药基因和可移动遗传元件的检测,评估水产微生物制剂源细菌的耐药性,为水产养殖中益生菌的筛选和安全应用提供科学依据。结果显示,分离得到的211株细菌可分为9个属,其中以芽孢杆菌属、肠球菌属、乳杆菌属为主。药敏实验结果显示,211株分离菌株中98株(46.7%)细菌表现为耐药。进一步对耐药性较强和多重耐药的79株细菌进行五大类(磺胺类、氨基糖苷类、四环素类、大环内酯类和β-内酰胺类)耐药基因和可移动遗传元件检测。结果显示,79株耐药菌中携带耐药基因的菌株阳性率为74.7%,存在多重耐药基因的菌株高达40.5%,氨基糖苷类和磺胺类耐药基因的检出率最高,分别为53.2%和40.5%。可移动遗传元件(Tn 916/1545)检出率为13.9%。综上可知,市售微生物制剂中发现了耐药菌株以及耐药基因,表明水产用微生物制剂存在一定的安全隐患,应加强对微生物制剂的安全性检测和风险评估。Microbial agents have developed rapidly in aquaculture because of their non-toxicity,pollution-free and low cost advantages,which have become the most potential products to replace antibiotics.However,the safety of microbial agents is difficult to guarantee.The presence of drug-resistant bacteria and antibiotics resistance genes(ARGs)in microbial agents can also potentially threaten the aquatic animal and human health.The drug resistance genes and mobile genetic elements were detected to isolate,identify the bacteria from the commercially available aquatic microbial agents,in addition to investigating the drug resistance of the isolates obtained from these preparations based on the antibiotic sensitivity test.Furthermore,the studies were of great significance to guiding the production of healthy microbial agents.They will provide a scientific basis for the evaluation and security application of probiotics in aquaculture.The agar dilution method was used to screen 45 kinds of aquatic microbial agents purchased on the market under sterile conditions,the sequence analysis of 16S rDNA,gyrB and construction of phylogenetic tree were used to identify isolates,the isolates from microbial agents were tested by Kirby-Bauer disk diffusion(K-B)method for their susceptibility to 11 different antibiotics(chloramphenicol,enrofloxacin,roxithromycin,sulfamethoxazole,streptomycin,tetracycline,gentamicin,penicillin,florfenicol,vancomycin,cephalexin),79 strains(28 strains were Bacillus,28 strains were Enterococcus,6 strains were Lactobacillus,3 strains were Acetobacter,2 strains were Pediococcus,1 strain was Plesiomonas,and 1 strain was Enterobacter)with significant drug resistance and multiple drug resistance were tested by Polymerase Chain Reaction(PCR)and sequence analysis of positive products for detecting mobile genetic elements(int1,int2,and Tn 916/1514)and five types of drug resistance genes(sulfonamides,aminoglycosides,tetracyclines,macrolides,andβ-lactams).The results showed that 211 strains were isolated from microbial a

关 键 词：微生物制剂 分离 鉴定 耐药性 抗性基因 

分 类 号：S917[农业科学—水产科学]