锯缘青蟹混合感染症致病菌的分离鉴定与感染治疗

doi:10.11978/j.issn.1009-5470.2010.05.103

热带海洋学报 ›› 2010, Vol. 29 ›› Issue (5): 103-110.doi: 10.11978/j.issn.1009-5470.2010.05.103cstr: 32234.14.j.issn.1009-5470.2010.05.103

锯缘青蟹混合感染症致病菌的分离鉴定与感染治疗

夏小安1, 吴清洋1, 李远友1, 王树启1, 游翠红1, 林尤顺2

1. 汕头大学广东省海洋生物技术重点实验室, 广东汕头515063; 2. 汕头市大顺实业有限公司, 广东汕头515000

收稿日期:2008-08-04 修回日期:2009-12-07 出版日期:2010-10-30 发布日期:2011-10-10
通讯作者: 李远友。
作者简介:夏小安(1982—), 男, 江西省新余市人, 硕士, 主要从事水产动物病害学研究。E-mail: xia1982817@163.com
基金资助:
广东省科技计划重大专项(2006A36502004)和农业攻关项目(2008B020800004)；广东高校产学研结合示范基地科技成果转化重
大项目(cgzhzd0812)；广东省教育部产学研合作专项资金项目(2007B090400049)；广东省海洋渔业科技推广专项(A200905J01)

Isolation and identification of two bacterial pathogens from mixed infection mud crab Scylla serrata and drug therapy

XIA Xiao-an1, WU Qing-yang1, LI Yuan-you1, WANG Shu-qi1, YOU Cui-hong1, LIN You-shun2

1. Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou 515063 China; 2. Shantou Dashun In-dustry Co., Limited, Shantou 515000 China

Received:2008-08-04 Revised:2009-12-07 Online:2010-10-30 Published:2011-10-10
Contact: 李远友。
About author:夏小安(1982—), 男, 江西省新余市人, 硕士, 主要从事水产动物病害学研究。E-mail: xia1982817@163.com
Supported by:
广东省科技计划重大专项(2006A36502004)和农业攻关项目(2008B020800004)；广东高校产学研结合示范基地科技成果转化重
大项目(cgzhzd0812)；广东省教育部产学研合作专项资金项目(2007B090400049)；广东省海洋渔业科技推广专项(A200905J01)

摘要/Abstract

摘要：

从汕头市牛田洋水产养殖区的患病锯缘青蟹Scylla serrata中分离纯化到2株优势细菌, 在TCBS培养基上生长, 颜色呈黄色的命名为HY, 呈绿色的命名为HG, 两者都是革兰氏阴性杆菌。人工感染此两株细菌的健康青蟹均产生与自然病蟹相同的症状和组织病理变化, 并导致死亡, 说明它们都是青蟹的病原菌。以这两种细菌的DNA为模板, 采用细菌16S rRNA基因的通用引物进行PCR扩增, 测序后分别获得HY 的1509bp和HG 的1532bp核苷酸序列。通过基因序列同源性比对和系统发育树分析, 结合细菌生理生化特性分析, HY和HG分别被鉴定为嗜水气单胞菌Aeromonas hydrophila和副溶血弧菌Vibrio parahaemolyticus。这是首次报道嗜水气单胞菌为青蟹的病原菌。两种病原菌均对阿莫西林、氧氟沙星、庆大霉素、链霉素高度敏感, 对磺胺嘧啶、甲砜霉素中度敏感, 对青霉素钠不敏感。采用嗜水气单胞菌浸泡感染或注射感染青蟹, 发病后进行阿莫西林药浴3天, 均获得100%的治疗效果。

关键词: 锯缘青蟹Scylla serrata, 嗜水气单胞菌Aeromonas hydrophila, 副溶血弧菌Vibrio parahaemolyticus, 组织病理学, 病原分离与鉴定, 药物防治

Abstract:

Two preponderant bacterial strains were isolated from the infected mud crab (Scylla serrata) in Niutianyang aqua-culture area of Shantou. One displayed yellow color and the strain was named as HY, the other showed green color and the strain was named as HG after growing on the TCBS agar plate. Both of them were rod shape and displayed gram negative. After healthy S. serrata individuals were infected with HY or HG by intramuscular injection, obvious histopathological changes could be observed in their tissues as those in the naturally diseased S. serrata, which lead them to their death. These results demonstrate that both HY and HG are pathogens of S. serrata. With the total HY or HG DNA as a template and bacte-rial 16S rRNA gene universal primers as primers, PCR amplification could obtain a partial HY sequence with 1509bp and HG fragment with 1532bp. Based on the blasting of 16S rRNA gene sequence and phylogenetic tree, together with the physiologi-cal and biochemical characteristics analysis, HY and HG were identified as Aeromonas hydrophila and Vibrio parahaemolyti-cus, respectively. To our knowledge, this is the first report that A. hydrophila is a pathogen of S. serrata. Both HY and HG showed high sensitivity to Ofloxacin, Gentamicin, streptomycin and Amoxicillin, medium sensitivity to Sulfadiazine and Thiamphenicol, and no sensitivity to Benzylpenicillin Sodium. After the healthy mud crabs were infected with A. hydrophila by immersion or intramuscular injection and displayed typical symptoms, drug therapy shows 100% success rate after the infected crabs were bathed in Amoxicillin at 15μg/mL for three days.

Key words: Scylla serrata, Aeromonas hydrophila, Vibrio parahaemolyticus, histopathology, isolation and identification of pathogens, drug therapy

夏小安,吴清洋,李远友,王树启,游翠红,林尤顺. 锯缘青蟹混合感染症致病菌的分离鉴定与感染治疗[J]. 热带海洋学报, 2010, 29(5): 103-110.

XIA Xiao-an,WU Qing-yang,LI Yuan-you,WANG Shu-qi,YOU Cui-hong,LIN You-. Isolation and identification of two bacterial pathogens from mixed infection mud crab Scylla serrata and drug therapy[J]. Journal of Tropical Oceanography, 2010, 29(5): 103-110.

参考文献

[1] 徐海圣, 舒妙安, 占秀安, 等. 锯缘青蟹常见病害及其防治技术[J]. 水产科学, 2000, 19 (5): 24-26.

[2] 毛芝娟, 卓华龙, 季芳, 等. 锯缘青蟹细菌性传染病的病原菌研究[J]. 台湾海峡, 2001, 20 (2): 187-192.

[3] 黄琪琰. 水产动物疾病学[M]. 上海: 上海科学技术出版社, 1993, 139 -141.

[4] 吴天靖, 丁木湖. 青蟹与梭子蟹主要疾病与防治[J]. 科学养鱼, 1998, (7): 26-27.

[5] 冯兴钱, 方家仲. 青蟹养殖技术[M]. 杭州: 浙江科学技术出版社, 1996, 206-215.

[6] 许文军, 施慧, 徐广庆, 等. 锯缘青蟹黄水病病原研究及综合防控初报[J]. 中国水产, 2006, 8: 54-56.

[7] 许文军, 绳秀珍, 徐汉祥, 等. 血卵涡鞭虫在养殖锯缘青蟹中的寄生[J]. 中国海洋大学学报, 2007, 37 (6): 916-920.

[8] 宋振荣, 倪子绵, 霍振华, 等. 锯缘青蟹肌肉组织坏死的病理研究[J]. 集美大学学报:自然科学版 , 2003, 8 (4): 301-304.

[9] 张叔勇, 郑大胜, 白志强, 等. 一株锯缘青蟹呼肠孤病毒的分离纯化及鉴定[J]. 海洋渔业, 2007, 29 (3): 277-280.

[10] 邹清, 秦骥, 朱小明. 锯缘青蟹一种球状病毒的分离和组织病理观察[J]. 厦门大学学报:自然科学版, 2007, 46 (6): 884-888.

[11] LI YY, XIA XA, WU QY, et al. Infection with Hematodinium sp. in mud crabs Scylla serrata cultured in low salinity water in southern China[J]. Diseases of Aquatic Organisms, 2008, 82: 145-150.

[12] MOORE R J, SHEEDY S A, DORAM T J, et al. Improved vector for expression library immunization-application to Mycoplasma hyopneumoniae infection in pig [J]. Vaccine, 2002, 20: 115-120.

[13] POLZ M F, CAVANAUGH C M. Bias in Template-to-Product Ratios in Multitemplate PCR [J]. Appl Environ Microbiol, 1998, 64 (10): 3724-3730.

[14] 莫照兰, 茅云翔, 陈师勇, 等. 养殖牙鲆鱼苗腹水症病原菌的鉴定及系统发育学分析[J]海洋与湖沼, 2003, 34 (2): 131-141.

[15] 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: 106-116.

[16] 陈昌福, 王敏. 嗜水气单胞菌对各类抗生素的敏感性测定[J]. 淡水渔业, 1996, 26 (6): 3-6.

[17] 倪语星, 洪秀华. 细菌耐药感染性监测与抗感染治疗[M]. 北京: 人民军医出版社, 2002: 83-85.

[18] AUSTIN B. Bacterial pathogens of fish [J]. Applied Bacteriology, 1995, 38: 483-506.

[19] 陈翠珍, 张晓君, 房海, 等. 鲤鱼体表出血感染症的病原菌检验[J]. 河北科技师范学院学报, 1998, 12 (1): 10-14.

[20] 张晓君, 陈翠珍, 何振平, 等. 草鱼细菌性烂鳃病的检验[J]. 中国农学通报, 1998, 14 (1): 45-46.

[21] 陆宏达, 金丽华, 范丽萍, 等. 中华绒螯蟹细菌性病原的分离和鉴定[J]. 水产学报, 1999, 23 (4): 381-386.

[22] 李文宽, 于翔, 闻秀荣, 等. 辽宁地区河蟹暴发性流行病病原研究[J]. 水产科学, 1999, 18 (4): 10-13.

[23] 杨先乐, 蔡完其. 中华绒螯蟹“抖抖病”流行情况初步调查[J]. 水产科技情报, 1998, 25 (6): 278-279

[24] 纪荣兴, 黄少涛. 锯缘青蟹“黄体病”病原菌的研究[J]. 台湾海峡, 1998, 17 (4): 473-476.

[25] 纪荣兴, 黄少涛, 吴志岭. 锯缘青蟹一种细菌病致病菌的研究[C]//中国科学技术协会学会部. 第二届全国人工养殖对虾疾病综合防治和环境管理学术研讨会论文集. 青岛: 海洋大学出版社, 1996: 119-123.

[26] 毛芝娟, 卓华龙. 锯缘青蟹细菌性传染病的病原菌研究[J]. 水产科学, 2000, 20 (1): 8-11.

[27] 周凯, 来琦芳. 中国明对虾弧菌病的组织病理学研究[J]. 海洋渔业, 2005, 27 (1): 49-54.

[28] 陶保华, 胡超群, 任春华. 斑节对虾弧菌病的病理学研究[J]. 热带海洋学报, 2001, 20 (3): 47-52.

[29] CLAIRE L V, CAROLINA C R, ANDREW F R. A histological study of shell disease syndrome in the edible crab Cancer pagurus [J]. Diseases of Aquatic Organisms, 2001, 47: 209-217.

锯缘青蟹混合感染症致病菌的分离鉴定与感染治疗

Isolation and identification of two bacterial pathogens from mixed infection mud crab Scylla serrata and drug therapy

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