黑斑蛙蝌蚪出血病病原菌的分离鉴定及药物敏感性  

Isolation, identification and drug sensitivity of the pathogen causing hemorrhagic disease of Rana nigromaculata tadpole

作　　者：钟为铭 彭芳 陈康勇 赵天宇 邹钧 高志鹏[1,2] ZHONG Weiming;PENG Fang;CHEN Kangyong;ZHAO Tianyu;ZOU Jun;GAO Zhipeng(College of Animal Science and Technology,Hunan Agricultural University,Changsha 410128,China;Hunan Engineering Technology Research Center of Featured Aquatic Resources Utilization,Changsha 410128;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306,China;College of Horticulture,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学动物科学技术学院,湖南长沙410128 [2]湖南省特色水产资源利用工程技术研究中心,湖南长沙410128 [3]上海海洋大学水产与生命学院,上海201306 [4]湖南农业大学园艺学院,湖南长沙410128

出　　处：《水产学报》2022年第1期107-115,共9页Journal of Fisheries of China

基　　金：国家自然科学基金(32073020);湖南省教育厅科学研究重点项目(20A238);国家级大学生创新训练计划(s202010537005)。

摘　　要：为确定黑斑蛙蝌蚪出血病的病原菌并筛选出防治该病的药物,本研究从患出血病蝌蚪体内分离出菌株KD-CXB-1,健康蝌蚪在人工感染该菌后出现与自然感染蝌蚪相似的出血症状。同时从人工感染死亡蝌蚪体内分离到与菌株KD-CXB-1形态特征及生理生化特性一致的优势菌株,以上结果表明KD-CXB-1为黑斑蛙蝌蚪出血病的病原菌。实验进一步对菌株KD-CXB-1开展形态学观察、革兰氏染色、生理生化指标测定、16S rDNA基因序列测定、系统发育树构建、药物敏感性测定。形态学观察显示该菌为杆状的革兰氏阴性菌,长度约1μm,表面粗糙。通过16S rDNA基因序列比对、系统发育树构建、生理生化实验确定KD-CXB-1为嗜水气单胞菌。药物敏感性实验结果表明,KD-CXB-1对氟苯尼考、丁香和苏木等10种药物高度敏感;对乌梅、五倍子和黄芩等3种药物中度敏感;对阿莫西林、大黄和青霉素等8种药物不敏感。研究表明,嗜水气单胞菌是引起黑斑蛙蝌蚪出血病的病原菌,筛选出的10种高敏感药物为有效防治蝌蚪出血病提供了理论参考和实践依据。To identify the pathogen causing hemorrhagic disease of Rana nigromaculata tadpoles and screen out sensitive drugs to control the disease, in this study, a strain KD-CXB-1 was isolated from diseased tadpoles, and healthy tadpoles showed hemorrhagic symptoms similar to those of naturally infected ones after being artificially infected with strain KD-CXB-1. Meanwhile, a dominant strain consistent with the morphological, physiological and biochemical characteristics of strain KD-CXB-1 was isolated from the dead tadpoles artificially infected.These results indicated that strain KD-CXB-1 was the pathogen causing hemorrhagic disease of R. nigromaculata tadpoles. Moreover, based on strain KD-CXB-1, several experiments were performed including morphological observation, gram staining, physiological and biochemical tests, 16 S rDNA gene sequencing, construction of phylogenetic trees, and drug susceptibility. The morphological observation showed that the KD-CXB-1 was a rodshaped Gram-negative bacterium with the length of about 1 μm and rough surface. Strain KD-CXB-1 was identified as Aeromonas hydrophila by 16 S rDNA sequencing and physiological and biochemical tests. Drug susceptibility tests indicated that strain KD-CXB-1 was highly sensitive to 10 drugs such as florfenicol, Syzygium aromaticum and Caesalpinia sappan;moderately sensitive to Prunus mume, Rhus chinensis and Scutellaria baicalensis, and insensitive to 8 drugs such as amoxicillin, Rheum palmatum and penicillin. In conclusion, our study proved that A. hydrophila was the pathogen causing hemorrhagic disease of R. nigromaculata tadpoles, and the highly sensitive drugs screened out by drug susceptibility tests may provide theoretical reference and practical basis for the effective control of hemorrhagic disease of R. nigromaculata tadpoles.

关 键 词：黑斑蛙 蝌蚪 嗜水气单胞菌 病原鉴定 药物敏感性 

分 类 号：S947.2[农业科学—水产养殖]