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Detection of Actinobacillus pleuropneumoniae in drinking water from pig farms

机译:猪场饮用水中胸膜肺炎放线杆菌的检测

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Actinobacillus pleuropneumoniae is the aetiological agent of porcine pleuropneumonia and is normally transmitted by aerosols and direct contact between animals. A. pleuropneumoniae has traditionally been considered an obligate pathogen of pigs and its presence in the environment has yet to be investigated. Here, the presence of A. pleuropneumoniae was detected in drinking water of pig farms in Mexico using a PCR specific for the RTX toxin gene, apxIV. The presence of A. pleuropneumoniae in farm drinking water was confirmed by indirect immunofluorescence using an A. pleuropneumoniae-specific polyclonal antibody and by fluorescent in situ hybridization. Viable bacteria from the farm drinking water were detected using the Live/Dead BacLight stain. Additionally, viable A. pleuropneumoniae was selected and isolated using the cAMP test and the identity of the isolated bacteria were confirmed by Gram staining, a specific polyclonal antibody and an A. pleuropneumoniae-specific PCR. Furthermore, biofilms were observed by scanning electron microscopy in A. pleuropneumoniae-positive samples. In conclusion, our data suggest that viable A. pleuropneumoniae is present in the drinking water of swine farms and may use biofilm as a strategy to survive in the environment.
机译:胸膜肺炎放线杆菌是猪胸膜肺炎的病原体,通常通过气溶胶和动物之间的直接接触传播。胸膜肺炎链球菌传统上被认为是猪的专性病原体,其在环境中的存在尚待调查。在这里,使用针对RTX毒素基因apxIV的特异性PCR在墨西哥养猪场的饮用水中检测到胸膜肺炎链球菌。通过使用胸膜肺炎链球菌特异性多克隆抗体的间接免疫荧光法和通过荧光原位杂交,证实了农场饮用水中胸膜肺炎链球菌的存在。使用活/死BacLight染色剂检测了农场饮用水中的活细菌。此外,选择活的胸膜肺炎链球菌,并使用cAMP测试进行分离,并通过革兰氏染色,特异性多克隆抗体和胸膜肺炎链球菌特异性PCR确认分离出的细菌的身份。此外,通过扫描电子显微镜在胸膜肺炎链球菌阳性样品中观察到生物膜。总之,我们的数据表明,猪场的饮用水中存在可行的胸膜肺炎放线杆菌,并可能将生物膜用作在环境中生存的策略。

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