黄鳝病原性维氏气单胞菌温和生物变种的分离与鉴定

摘要/Abstract

摘要： 从养殖患出血病黄鳝(Monopterus albus)的肝脾肾中分离到大量菌落形态和色泽一致的细菌。16S rRNA序列分析证实随机挑选的5个分离株为同一种细菌，其中一株编号为HS120920进一步鉴定。结合细菌的形态特征，生理生化特征及16S rRNA和gyrB序列分析结果，鉴定该菌为维氏气单胞菌温和生物变种(Aeromonas veronii biovar sobria)。该菌对黄鳝的半致死浓度约为5.79×10⁷CFU/mL。药敏试验结果显示，菌HS120920对7种喹诺酮类，5种氨基糖苷类等17种抗菌药物高度敏感，对其他3种抗菌药物低敏，或有抗性。

关键词: 黄鳝, 出血病, 维氏气单胞菌, 16S, rRNA基因, gyrB基因

Abstract: Large colonies with the same morphology and color were isolated from liver, spleen and kidney of diseased mud eel, Monopterus albus. 5 isolates randomly selected were proved to be the same bacteria by 16S rRNA sequence analysis. Further research of one strain named HS120920 among them was developed. According to morphology, physiological and biochemical characteristics, analysis of their 16S rRNA and gyrB sequences, the strain was identified as Aeromonas veronii biovar sobria, the LD50 was 5.79×10⁷ CFU/mL. The susceptibility test showed that the strain was highly sensitive to 17 drugs such as seven quinolones, five aminoglycosides, but showed low sensitivity or resistance to other 3 drugs.

Key words: Monopterus albus ,Hemorrhagic disease, Aeromonas veronii , 16S, rRNA gene, GyrB gene

陈红莲, 江河, 胡王, 凌俊, 段国庆. 黄鳝病原性维氏气单胞菌温和生物变种的分离与鉴定[J]. 生物技术通报, 2014, 0(3): 130-136.

Chen Honglian, Jiang He, Hu Wang, Ling Jun, Duan Guoqing. Isolation and Identification of Pathogenic Aeromonas veronii Biovar Sobria from Monopterus albus[J]. Biotechnology Bulletin, 2014, 0(3): 130-136.

参考文献

[1] 陈怀青, 陆承平. 嗜水气单胞菌:黄鳝出血性败血症的病原[J].中国人兽共患病杂志, 1991, 7(4):21-23.
[2] 沈锦玉, 刘问, 钱冬. 养殖黄鳝出血病病原的研究[J].浙江海洋学院学报:自然科学版, 2001, 20(2):120-122.
[3] 彭彬, 杨光友, 陈晓利, 等. 黄鳝出血病病原菌的分离与鉴定[J]. 淡水渔业, 2011, 41(3):72-76.
[4] 舒妙安, 马有智.黄鳝产碱假单胞菌的分离与鉴定[J]. 淡水渔业, 2000, 30(5):33-34.
[5] 徐海圣, 舒妙安.黄鳝温和气单胞菌病病原的分离鉴定及免疫应答[J].中国兽医学报, 2003, 23(3):240-242.
[6] 任红梅, 何智, 杨德英, 等.黄鳝体表溃疡病病原的分离与鉴定[J].水生生物学报, 2010, 34(6):5-11.
[7] 彭彬, 杨光友, 陈晓利, 等.黄鳝藤黄微球菌的分离鉴定及药敏试验[J].上海海洋大学学报, 2011, 20(3):405-411.
[8] Reed LJ, Muech H. A simple method of estimating fifty percent endpoints[J]. Am J Hyg, 1938, 27:493-497.
[9] 东秀珠, 蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社, 2001:364-398.
[10] Brenner DJ, Krieg NR, Staley JT. Bergey’s manual of systematic bacteriology(second edition Part B)[M]. Dordrecht Heidelberg London & New York:Springer-verlag, 2005:557-578.
[11] CLSI. Performance standards for antimicrobial susceptibility tes-ting；twenty third informational supplement[J]. CLSI document M100-S23. Wayne, PA:Clinical and laboratory standards institute, 2013, 33(1):45-128.
[12] Olsen GJ, Pace NR, Nuell M, et al. The ribosomal RNA database pro-ject[J]. Nucleic Acids Res, 1991, 19(Suppl.):2017-2021.
[13] 侯晓丽, 陈智.分类及鉴别细菌的新靶标-gyrB基因[J].国外医学?流行病学传染病学分册, 2005, 32(1):38-41.
[14] 郝云婕, 韩素贞.gyrB基因在细菌系统进化发育分析中的应用[J].生物技术通报, 2008(2):39-41.
[15] Yá?ez MA, Catalán V, Apráiz D, et al. Phylogenetic analysis of members of the genus Aeromonas based on gyrB gene sequences[J]. Int J Syst Evol Microbiol, 2003, 53:875-883.
[16] Küpfer M, Kuhnert P, Korczak BM, et al. Genetic relationships of Aeromonas strains inferred from 16S rRNA, gyrB and rpoB gene sequences[J]. Int J Syst Evol Microbiol, 2005, 56:2743-2751.
[17] Gr?bner S, Bissinger AL, Raible A, et al. Severe diarrhoea caused by Aeromonas veronii biovar sobria in a patient with metastasised GIST[J]. Pol J Microbiol, 2007, 56(4):277-279.
[18] Mencacci A, Cenci E, Mazzolla R, et al. Aeromonas veronii biovar veronii septicaemia and acute suppurative cholangitis in a patient with hepatitis B[J]. J Med Microbiol, 2003, 52(8):727-30.
[19] Sánchez-Cáspedes J, Figueras MJ, Aspiroz C, et al. Development of imipenem resistance in an Aeromonas veronii biovar sobria clinical isolate recovered from a patient with cholangitis[J]. J Med Microbiol, 2009, 58:451-455.
[20] Cui H, Hao S, Arous E. A distinct cause of necrotizing fasciitis: Aeromonas veronii biovar sobria[J]. Surgical Infections, 2007, 8(5):523-528.
[21] Steinfeld S, Rossi C, Bourgeois N. Septic arthritis due to Aeromonas veronii biotype sobria[J]. Clin Infect Dis, 1998, 27:402-403.
[22] Shiina Y, Li K, Iwanaga M. An Aeromonas veronii biovar sobria infection with disseminated intravascular gas production[J]. J Infect Chemother, 2004, 10(1):37-41.
[23] Rahman M, Colque-Navarro P, Kühn I, et al. Identification and characterization of pathogenic Aeromonas veronii biovar sobria asso-ciated with epizootic ulcerative syndrome in fish in Bangladesh[J]. Appl Environm Microbiol, 2002, 68(2):650-655.
[24] Cai SH, Wu ZH, Jian JC. Characterization of pathogenic Aeromonas veronii bv. veronii associated with ulcerative syndrome from chinese longsnout catfish(Leiocassis longirostris günther)[J]. Braz J Microbiol, 2012, 43(1):382-388.
[25] Zheng WD, Cao HP, Yang XL. Aeromonas veronii infection in the cultured snakehead fish, Ophiocephalus argus(Cantor)[J]. Afr J Microbiol Res, 2012, 6(44):7218-7223.
[26] 唐毅, 郑凯迪, 朱成科, 等.华鲮烂尾病病原的分离鉴定及药敏分析[J].淡水渔业, 2010, 40(4):50-55.
[27] 沈文英, 李卫芬, 高研.中华鳖病原性维隆气单胞菌的分离及药敏试验[J].水生生物学报, 2010, 34(5):1060-1064.
[28] 夏飞, 梁利国, 谢骏.异育银鲫病原维氏气单胞菌的分离鉴定及药敏试验[J].淡水渔业, 2012, 42(5):22-26.
[29] 秦蕾, 徐静, 张晓君.泥鳅的凡隆气单胞菌感染[J].中国人畜共患病学报, 2008, 24(12):1000-1002.
[30] 龚倩, 高淑琴, 单晓枫, 等.框镜鲤致病性维氏气单胞菌的分离鉴定[J].中国预防兽医学报, 2010, 32(12):981-983.
[31] 杨求华, 郭松林, 关瑞章, 等.鳗鲡病原性维氏气单胞菌分离与鉴定[J].生物技术通报, 2012(7):134-139.
[32] 潘晓艺, 沈锦玉, 李建应, 等.青虾“软壳综合症”病原及其特性[J].微生物学通报, 2009, 36(10):1571-1576.
[33] 马志宏, 杨慧, 李铁梁.西伯利亚鲟(Acipenser baerii)致病性维氏气单胞菌的分离鉴定[J].微生物学报, 2009, 49(10):1289-1294.