中华绒螯蟹养殖用抑水绵枯草芽孢杆菌A4的安全性分析  

Safety analysis of Bacillus subtilis A4 against Spirogyra in Eriocheir sinensis aquaculture use

作　　者：曹海鹏[1,2] 顾颖 孙苗苗 王会聪[4] 叶仁智 安健 CAO Hai-peng;GU Ying;SUN Miao-miao;WANG Hui-cong;YE Ren-zhi;AN Jian(National Pathogen Collection Center for Aquatic Animals,Shanghai Ocean University,Shanghai 201306,China;Shanghai Engineering Research Center for Aquaculture,Shanghai Ocean University,Shanghai 201306,China;Marine and Fisheries Development Promotion Center of Lianyungang City,Lianyungang 222000,Jiangsu,China;Jiangsu Vocational Agriculture and Forestry College,Jurong 212400,Jiangsu,China)

机构地区：[1]上海海洋大学,国家水生动物病原库,上海201306 [2]上海海洋大学,上海水产养殖工程技术研究中心,上海201306 [3]连云港市海洋与渔业发展促进中心,江苏连云港222000 [4]江苏农林职业技术学院,江苏句容212400

出　　处：《淡水渔业》2023年第5期104-112,共9页Freshwater Fisheries

基　　金：国家虾蟹产业技术体系专项资金(CARS-48);江苏农林职业技术学院科技计划项目(2022kj58)。

摘　　要：为评价中华绒螯蟹(Eriocheir sinensis)养殖用抑水绵枯草芽孢杆菌(Bacillus subtilis)A4的安全性,本实验从溶血性、有害代谢物产生能力、耐药性与耐药质粒及其对斑马鱼、大型蚤、中华绒螯蟹的毒性等方面对枯草芽孢杆菌A4的安全性进行了检测分析。实验结果表明,枯草芽孢杆菌A4无溶血活性,不产生生物胺、吲哚、氨、硫化氢等有害代谢物,不携带nheA、bceT、ces、cytK、entFM、hblA、hblC和hblD等毒力基因,对头孢菌素类、β-内酰胺类、氨基糖苷类、四环素类、大环内酯类、喹诺酮类、多肽类、硝基呋喃类、酰胺醇类等抗生素高度敏感,不携带MCR-1、floR、aac(6′)-Ⅰb、bla_(CTX)、ermA、ermC、qnrA、qnrB、sul1、sul2、tetA和tetM等耐药基因且无耐药质粒。此外,枯草芽孢杆菌A4对斑马鱼、大型蚤和中华绒螯蟹的半数致死浓度>1.5×10^(9)CFU/mL。本研究表明枯草芽孢杆菌A4在中华绒螯蟹养殖中具有良好的应用安全性。the safety of Bacillus subtilis A4 was assessed by its hemolysis,toxic and harmful metabolite production,antibiotic resistance and antibiotic-resistant plasmid as well as the toxicity to zebrafish,Daphnia magna and Chinese mitten crab assays to evaluate the safety of B.subtilis A4 in Eriocheir sinensis culture use against Spirogyra.The results showed that B.subtilis A4 had no hemolytic activity,which did not produce harmful metabolites such as biogenic amines,indole,ammonia,and hydrogen sulfide,and did not carry virulent nheA,bceT,ces,cytK,entFM,hblA,hblC,and hblD genes.In addition,B.subtilis A4 was highly sensitive to cephalosporins,β-lactams,aminoglycosides,tetracyclines,macrolides,quinolones,peptides,nitrofurans,and amide alcohols antibiotics,which did not possess neither antibiotic-resistant genes like MCR-1,floR,aaac(6′)-Ib,bla_(CTX),erm A,erm C,qnr A,qnr B,sul1,sul2,tetA and tetM nor resistance plasmid.Its median lethal concentrations to zebrafish,D.magna and E.sinensis was tested to be above 1.5×10^(9) CFU/mL.The present study demonstrates B.subtilis A4 showed good safety in Chinese mitten crab culture.

关 键 词：枯草芽孢杆菌(Bacillus subtilis) 水绵 安全性 中华绒螯蟹(Eriocheir sinensis) 

分 类 号：S966.16[农业科学—水产养殖]