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任红梅, 何智, 杨德英, 杨光友, 古小彬, 王淑贤. 黄鳝体表溃疡病病原菌的分离与鉴定[J]. 水生生物学报, 2010, 34(6): 1106-1112. DOI: 10.3724/SP.J.1035.2010.01106
引用本文: 任红梅, 何智, 杨德英, 杨光友, 古小彬, 王淑贤. 黄鳝体表溃疡病病原菌的分离与鉴定[J]. 水生生物学报, 2010, 34(6): 1106-1112. DOI: 10.3724/SP.J.1035.2010.01106
REN Hong-Mei, HE Zhi, YANG De-Ying, YANG Guang-You, GU Xiao-Bin, WANG Shu-Xian. ISOLATION, IDENTIFICATION AND PATHOGENICITY OF CAUSATIVE PATHOGEN FOR MUSCULAR ULCER IN MONOPTERUS ALBUS[J]. ACTA HYDROBIOLOGICA SINICA, 2010, 34(6): 1106-1112. DOI: 10.3724/SP.J.1035.2010.01106
Citation: REN Hong-Mei, HE Zhi, YANG De-Ying, YANG Guang-You, GU Xiao-Bin, WANG Shu-Xian. ISOLATION, IDENTIFICATION AND PATHOGENICITY OF CAUSATIVE PATHOGEN FOR MUSCULAR ULCER IN MONOPTERUS ALBUS[J]. ACTA HYDROBIOLOGICA SINICA, 2010, 34(6): 1106-1112. DOI: 10.3724/SP.J.1035.2010.01106

黄鳝体表溃疡病病原菌的分离与鉴定

ISOLATION, IDENTIFICATION AND PATHOGENICITY OF CAUSATIVE PATHOGEN FOR MUSCULAR ULCER IN MONOPTERUS ALBUS

  • 摘要: 研究首次报道从患体表溃疡病黄鳝的肝脏、肾脏、脾脏等分离到1株细菌, 经动物回归试验鉴定其为病原菌。形态学观察、生理生化试验和16S rRNA基因鉴定, 结果证实该病原菌株为温和气单胞菌(Aeromonas sobria)。通过致病性试验证明其对鲤鱼和泥鳅均有致病力;药敏试验结果表明:分离菌株对头孢氨噻肟、头孢曲松钠、丙氟哌酸、洛美沙星、甲氧苄啶、氟哌酸、壮观霉素等高度敏感,而对氨苄青霉素、乙酰螺旋霉素等药物不敏感。

     

    Abstract: Rice field eel, Monopterus albus, freshwater fish, belongs to the synbranchidae. It is an economically important species for fish production in China, especially in southern of China. The diseases of cultured rice field eel have been the bottle-neck for the sustainable development of rice field eel cultivation. The muscular ulcer disease is a kind of common disease to rice field eel and is suggested to be one of the most important diseases in industrial artificial-breeding. In 2007, the outbreak of muscular ulcer disease in the rice field eel farmed in Sichuan province, whichwas high incidence and mortality and caused lots of economical loss to the farmer. To make certain the causative pathogen, we studied the etiology and virulence of rice field eel so as to provide basic data for the farming industry of rice field eel. In the present study, we used the morphological, physiological and biochemical methods to identify the isolated bacterium. Meanwhile, to ensure the results accurately, the phylogenetic analyses were also used. Liver, kidney and spleen of diseased rice field eel were aseptically streaked on Nutrient Agar by swabbing. After incubation, pure cultured colonies were obtained and one causative bacteria (hs-1) was isolated from liver, kidney and spleen of moribund rice field eel. The characteristics of causative bacteria were tested by bacteria micro amount biochemical fermentation tube, Gram's staining and another method. The phylogenetic comparison based on 16S rRNA gene of the causative bacterium was also studied, the amplified nucleotide sequence of isolated bacterium (GenBank accession No. FJ461353) had 99% homology with the other 16S rRNA sequences of A.Sobria. Based on the morphological, physiological and biochemical characteristics, as well as the results of phylogenetic analysis, the causative bacterium was conducted as A.Sobria. Pathogenicity tests in rice field eel showed that the symptoms of rice field eel were similar with the natural diseased eels. The mortality of high dose injected group (3.0?08 CFU) was 100%. In all injected groups, the similar symptoms appeared in 48h post-artificial injection. The same pathogen was isolated from diseased fish caused by the artificial infection. The mortality of loach and carp in Pathogencity tests were 80% and 100%, which showed that the isolated strain also had pathogenicity to the others fish. A.Sobria was typically opportunistic pathogen and secondary invader. The strain in present study was resistant to much commonly antibiotics, such as Ampicillin, Acetylspiramycin, Novobiocin, and so on. The results of drug sensitivity test also showed that Cefotaxime Sodium, Rocephin, Norfloxacin, Ciprofloxacin, Lomefloxacin, Trimethoprim and Norfloxacin were most effective against the pathogenic bacterium.However, the treatment of A. hydrophila infections would be painstakingly tough and expensive. Therefore, the farmers should pay more attentions to cultural management, aquatic techniques and regulation.

     

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