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• Isolation and Identification of Aeromonas Veroni from Diseased Acrossocheilius Hemispinus ZHANG Jingjing Chinese Agricultural Science Bulletin    2023, 39 (20): 138-146.   DOI: 10.11924/j.issn.1000-6850.casb2022-0781 Abstract   （135） HTML （3） PDF（pc） （1802KB）（18）   The bacterial isolation, identification and drug sensitivity of diseased Acrossocheilius hemispinus with skin ulceration, surface hemorrhage, swollen and bleeding liver were studied. The bacterial strain was isolated and purified from liver tissue of the diseased fish and injected intramuscularly into healthy Acrossocheilius hemispinus. Full-length sequences of 16S rDNA and gyrB gene were obtained by PCR amplification of pathogenic bacteria, while physiological and biochemical identification, virulent genes detection, and drug sensitivity study were also conducted. The isolated strain was numbered as AS0829L1, and the artificial infection experiments showed that pathogenic groups of 5×1010, 5×109, 5×108 CFU/mL could cause 100%, 100%and 28.6% mortality, respectively. The BLAST result indicated the sequences of 16S rDNA and gyrB were homologous to the corresponding sequence of Aeromonas veronii in Genbank, with homology as high as 99.93% and 99.34%, respectively. Comprehensive molecular biology, physiological and biochemical characteristics, virulence gene detection results demonstrated the separate strain was Aeromonas veronii bv.sobria. The drug sensitivity test showed the isolated strain was highly sensitive to 20 antibacterial drugs including cefuroxime, cefoperazone, gentamicin, neomycin, enrofloxacin and ofloxacin, moderate sensitive to 5 antibacterial drugs including carboxybenzillin, streptomycin, flufenicol, and resistant to 12 antibacterial drugs including penicillin, norfloxacin, erythromycin. In this study, the pathogen of the diseased Acrossocheilius hemispinus was identified and the drug sensitivity test of the isolated strain was carried out, which provided a research basis for the prevention and control of diseases in aquaculture production. 药物 每片药量/μg 抑菌圈直径/mm 药物敏感性 药物 每片药量/μg 抑菌圈直径/mm 药物敏感性 β-内酰胺类 - - - 大环内酯类 - - - 青霉素 10U 0 R 红霉素 15 5 R 羧苄西林 100 21 I 罗红霉素 15 10 R 头孢曲松 30 46 S 麦迪霉素 30 0 R 头孢派酮 75 44 S 四环素类 - - - 头孢他啶 30 34 S 四环素 30 9 R 头孢呋辛 30 41 S 多西环素 30 9 R 头孢唑啉 30 19 S 氯霉素类 - - - 头孢氨苄 30 19 S 氟苯尼考 30 16 I 头孢拉定 30 20 S 氯霉素 30 20 S 氨基糖苷类 - - - 糖肽类 - - - 链霉素 10 13 - 万古霉素 30 13 R 卡那霉素 30 24 S 多粘菌素B 300IU 16 S 庆大霉素 10 16 S 林可酰胺类 - - - 新霉素 30 15 S 克林霉素 2 0 R 丁胺卡那 30 18 S 青霉素类 - - - 磺胺类 - - - 阿莫西林 20 9 R 复方新诺敏 23.75/1.25 24 S 苯唑西林 1 0 R 甲氧苄啶 5 20 S 派拉西林钠 100 25 S 喹诺酮类 - - - 硝基呋喃类 - - - 环丙沙星 5 29 S 呋喃唑酮 300 24 S 诺氟沙星 10 26 R 硝基米唑类抗生素 - - 恩诺沙星 10 23 R 甲硝唑 5 0 R 表4 分离菌株AS0829L1药敏试验结果 Other Images/Table from this Article 表1 毒力基因引物序列及扩增片段大小 图1 图2 表2 分离菌株AS0829L1回归实验结果 图3 A：体表发红、溃烂、鳞片脱落、烂尾（箭头）；B：肝脏发黄发白、脾脏发黑肿大（箭头） 图4 表3 分离菌株AS0829L1生理生化鉴定结果 图5 M-marker；1-act；2-ast；3-aer A；4-alt；5-hly A，6-tap A