余露军, 苗宗余, 蔡磊, 魏远征, 黄韧, 李建军. 诸氏鲻虾虎鱼“溃烂病”病原及其致病影响因子分析[J]. 南方水产科学, 2018, 14(5): 45-52. DOI: 10.3969/j.issn.2095-0780.2018.05.006
引用本文: 余露军, 苗宗余, 蔡磊, 魏远征, 黄韧, 李建军. 诸氏鲻虾虎鱼“溃烂病”病原及其致病影响因子分析[J]. 南方水产科学, 2018, 14(5): 45-52. DOI: 10.3969/j.issn.2095-0780.2018.05.006
YU Lujun, MIAO Zongyu, CAI Lei, WEI Yuanzheng, HUANG Ren, LI Jianjun. Pathogenicity and impact factors of pathogen causing ulcer disease on Mugilogobius chulae[J]. South China Fisheries Science, 2018, 14(5): 45-52. DOI: 10.3969/j.issn.2095-0780.2018.05.006
Citation: YU Lujun, MIAO Zongyu, CAI Lei, WEI Yuanzheng, HUANG Ren, LI Jianjun. Pathogenicity and impact factors of pathogen causing ulcer disease on Mugilogobius chulae[J]. South China Fisheries Science, 2018, 14(5): 45-52. DOI: 10.3969/j.issn.2095-0780.2018.05.006

诸氏鲻虾虎鱼“溃烂病”病原及其致病影响因子分析

Pathogenicity and impact factors of pathogen causing ulcer disease on Mugilogobius chulae

  • 摘要: 为研究诸氏鲻虾虎鱼(Mugilogobius chulae)“溃烂病”病原及其致病性,对患病鱼进行了病原分离、人工感染实验以及病原生理生化鉴定和gryB分析,研究了不同温度、盐度、pH条件对其致病性的影响。结果显示,病原菌为迟缓爱德华菌(Edwardsiella tarda),对诸氏鲻虾虎鱼和斑马鱼 (Danio rerio)的半致死剂量(LD50)分别为57 CFU·尾–1和2.0×104 CFU·尾–1,毒力基因(citCmukFesrBkatBfimAgadB)检测结果均为阳性,其基因序列与已知迟缓爱德华菌相应的毒力基因序列同源性高达99%。病理结果显示,患病鱼肾脏、肝脏、皮肤细胞均出现不同程度的变性、坏死和炎性细胞浸润等,其中肾和肝组织病变最为严重。25 ℃和30 ℃实验组5 d累计死亡率分别为(33±15)%、(60±10)%,显著高于20 ℃实验组和对照组 (P<0.05);盐度15和30实验组5 d累计死亡率分别为(60±10)%、(63±21)%,显著高于盐度3实验组 (20±10)% ,P<0.05;而pH 6.0、pH 7.5、pH 9.0实验组之间5 d累计死亡率无显著差异 (P>0.05)。结果表明,迟缓爱德华菌是诸氏鲻虾虎鱼“溃烂病”病原,具有强致病性,养殖温度和盐度对其致病性具有显著影响。

     

    Abstract: In order to investigate the pathogen and pathogenicity of ulcer disease of Mugilogobius chulae, we isolated the pathogenic bacteria from M.chulae for the artifical infection experiment, physiological and biochemical identification of pathogens and gryB analysis to determine if the environmental factors (temperature, salinity and pH) influence the pathogenicity of isolate strain EtMc1512. The results show that the pathogen was Edwardsiella tarda. The LD50s (lethal dose, 50%) of the isolate strain EtMc1512 to healthy M.chulae and Danio rerio were 57 CFU·ind–1 and 2.0×104 CFU·ind–1, respectively. The virulence-related genes (citC, mukF, esrB, katB, fimA, gadB) had been detected in strain EtMc1512, and the homology of their nucleotide between EtMc1512 and other E.tarda which were registered in GenBank was 99%. Cell degeneration, necrosis and inflammatory cell infiltration occurred with severe lesion in kidney and liver. The cumulative mortalities in high-temperature groups of 25 ℃ and 30 ℃ were significantly higher than those in low-temperature group of 20 ℃ and the control group (P<0.05). The cumulative mortality in low-salinity group of 3 was significantly lower than those in high-salinity groups of 15 and 30 (P<0.05). However, there was no significant difference in the cumulative mortality among different pH groups (6.0, 7.5 and 9.0, P>0.05). The results show that E.tarda is a highly pathogenic pathogen of ulcer disease of M.chulae, and the environmental factors of temperature and salinity affect the virulence of E.tarda significantly.

     

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