黄喉拟水龟肺气肿病病原菌的分离鉴定及其药敏特性分析
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摘要
【目的】明确黄喉拟水龟肺气肿病的病原菌及其药敏特性,为该病的临床诊断与治疗提供科学依据。【方法】通过微生物常规分离、人工感染试验、生理生化鉴定及16S rDNA序列分析等方法分离鉴定黄喉拟水龟肺气肿病的病原菌,测定该病原菌对黄喉拟水龟的半致死剂量(LD50),并采用K-B药敏纸片扩散法测定其药敏特性。【结果】从患肺气肿病黄喉拟水龟的肺脏组织中分离获得一株优势菌株(命名为GXNN1),其生理生化特性与肺炎克雷伯氏菌(Klebsiella pneumoniae)完全一致;经腹腔人工注射感染,黄喉拟水龟表现出与自然发病相同的症状,腹腔内有少量血水,肠壁充血,肺脏充血膨胀;菌株GXNN1对黄喉拟水龟的LD50为0.63×103CFU/g。菌株GXNN1与肺炎克雷伯氏菌的16S rDNA序列同源性为99.4%～99.9%,从基于16S rDNA序列同源性构建的系统发育进化树也发现,菌株GXNN1与肺炎克雷伯氏菌聚为一支。综合其形态特征、生理生化特性、人工感染试验及16S rDNA序列分析结果,可确定黄喉拟水龟肺气肿病的致病菌(菌株GXNN1)为肺炎克雷伯氏菌。药敏试验结果表明,菌株GXNN1对先锋必和舒普深敏感,对氨苄青霉素、氯霉素、多西环素、庆大霉素、诺氟沙星、阿莫西林等15种药物已产生耐药性(不敏感)。【结论】肺炎克雷伯氏菌是引起黄喉拟水龟肺气肿病的主要病原菌,LD50为0.63×103CFU/g,生产中可使用舒普深和先锋必等药物进行治疗。
【Objective】The pathogens and drug-sensitive properties of the Mauremys mutica pulmonary emphysema were clarified to provide scientific basis for the clinical diagnosis and treatment of the disease.【Method】The pathogens of M.mutica emphysema were isolated and identified using routine microbial isolation technology,artificial infection test,physiological and biochemical identification,16 S rDNA sequence analysis.The median lethal dose(LD50)of the pathogen to M.mutica was determined and the drug sensitivity characteristics of the pathogen were analyzed using Kirby-Bauer paper flakes diffusion method.【Result】A dominant strain(GXNN1)was isolated from the lung tissue of M.mutica with emphysema,physiological and biochemical reactions were consistent with Klebsiella pneumoniae.After intraperitoneal injection of GXNN1 to M.mutica,showing small amount of blood in abdominal cavity,intestinal wall hyperemia,lungs hyperemia and inflation occurred,which was the same symptoms as natural infection.The LD50 of GXNN1 to M.mutica was0.63×103CFU/g.The 16 S rDNA sequences homology between GXNN1 and K.pneumoniae were 99.4%-99.9%.Phylogenetic trees based on homology of 16 S rDNA sequences revealed that GXNN1 clustered with the same branch of K.pneumoniae.Synthetic morphological characteristics,physiological and biochemical characteristics,artificial infection test and 16 S rDNA sequence analysis results confirmed that the pathogen(GXNN1)causing M.mutica emphysema was K.pneumoniae.Drug sensitivity tests showed that GXNN1 isolate was highly sensitive to cefobid and sulperazone,and resistant(insensitive)to 15 antibiotics such as ampicillin,chloramphenicol,doxycycline,gentamicin,norfloxacin and amoxicillin.【Conclusion】K.pneumoniae is the main pathogen causing M.mutica with emphysema.The LD50 was 0.63×103CFU/g.Sulperazone and cefobid can be used for treatment in aquaculture production.
【Objective】The pathogens and drug-sensitive properties of the Mauremys mutica pulmonary emphysema were clarified to provide scientific basis for the clinical diagnosis and treatment of the disease.【Method】The pathogens of M.mutica emphysema were isolated and identified using routine microbial isolation technology,artificial infection test,physiological and biochemical identification,16 S rDNA sequence analysis.The median lethal dose(LD50)of the pathogen to M.mutica was determined and the drug sensitivity characteristics of the pathogen were analyzed using Kirby-Bauer paper flakes diffusion method.【Result】A dominant strain(GXNN1)was isolated from the lung tissue of M.mutica with emphysema,physiological and biochemical reactions were consistent with Klebsiella pneumoniae.After intraperitoneal injection of GXNN1 to M.mutica,showing small amount of blood in abdominal cavity,intestinal wall hyperemia,lungs hyperemia and inflation occurred,which was the same symptoms as natural infection.The LD50 of GXNN1 to M.mutica was0.63×103CFU/g.The 16 S rDNA sequences homology between GXNN1 and K.pneumoniae were 99.4%-99.9%.Phylogenetic trees based on homology of 16 S rDNA sequences revealed that GXNN1 clustered with the same branch of K.pneumoniae.Synthetic morphological characteristics,physiological and biochemical characteristics,artificial infection test and 16 S rDNA sequence analysis results confirmed that the pathogen(GXNN1)causing M.mutica emphysema was K.pneumoniae.Drug sensitivity tests showed that GXNN1 isolate was highly sensitive to cefobid and sulperazone,and resistant(insensitive)to 15 antibiotics such as ampicillin,chloramphenicol,doxycycline,gentamicin,norfloxacin and amoxicillin.【Conclusion】K.pneumoniae is the main pathogen causing M.mutica with emphysema.The LD50 was 0.63×103CFU/g.Sulperazone and cefobid can be used for treatment in aquaculture production.
引文
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