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首页> 外文期刊>Journal of neurosurgical sciences >Characterization of Virulent netB(+)/tpeL(+) Clostridium perfringens Strains from Necrotic Enteritis-Affected Broiler Chicken Farms
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Characterization of Virulent netB(+)/tpeL(+) Clostridium perfringens Strains from Necrotic Enteritis-Affected Broiler Chicken Farms

机译：毒性NetB（+）/ TPEL（+）梭菌的特征来自坏死肠炎影响肉鸡养鸡农场的菌株

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Clostridium perfringens (CP) type A and newly created type G strains are the key etiological factors in the induction of necrotic enteritis (NE), an important enteric disease that is responsible for the annual loss of $6 billion in the worldwide poultry industry. Several CP toxin genes were found to be critical in NE pathogenesis in chickens, but limited information is available on the CP lethal toxin tpeL gene. In this study, 19 CP strains isolated from NE-affected chicken farms were characterized microbiologically and molecularly and evaluated for their pathogenicity in commercial broiler chickens. Toxin typing by PCR revealed that all strains tested were positive for the netB toxin gene, but only five strains were positive for the tpeL toxin gene (LLY-TpeL 13, -TpeL 15, -TpeL 17, -TpeL 18, and -TpeL 19, simplified as TpeL 13, TpeL 15, TpeL 17, TpeL 18, and TpeL 19). High levels of TpeL proteins were detected in the concentrated culture supernatant from strains TpeL 13, 15, 17, and 19 by western blotting. Quantitative PCR showed that strains TpeL 13, 15, 17, 18, and 19 harbored a high number of copies of tpeL genes, while TpeL 18 had the highest number of copies of the tpeL gene among all CP strains tested when normalized with copy numbers of 16S rRNA gene as a housekeeping gene marker. The in vivo NE challenge test using multiple oral CP inoculations combined with a high-protein diet showed that TpeL 17 was the most virulent in inducing typical NE lesions, followed by TpeL 19 as the next most virulent, when tested in commercial broiler chickens. Infection with TpeL 17 reduced the growth rate significantly, as shown by reduced relative body weight gain percentage at day 5 postinfection. Availability of the virulent netB(+)tpeL(+) CP strains is essential for the development of a CP-alone NE challenge model that could provide significant tools for understanding CP pathogenesis and for development of alternative to antibiotics.
机译：流产硬螺栓（CP）类型A和新建的G株是诱导坏死肠炎（NE）的关键病因因素，这是一个重要的肠溶病，负责全球禽类工业的60亿美元。发现几种CP毒素基因在鸡的NE发病机制中是至关重要的，但是在CP致死毒素TPEL基因上可获得有限的信息。在本研究中，从Ne-Vised养鸡农场分离的19c株菌株进行了微生物学和分子，并在商业肉鸡鸡中进行了致病性。通过PCR键入的毒素显示，测试的所有菌株对于网状毒素基因是阳性的，但是对于TPEL毒素基因仅有五种菌株（Lly-TPEL 13，-TPEL 15，-TPEL 17，-TPEL 18和-TPEL 19 ，简化为TPEL 13，TPEL 15，TPEL 17，TPEL 18和TPEL 19）。通过蛋白质印迹，在菌株TPEL 13,15,17和19中浓缩培养上清液中检测到高水平的TPEL蛋白。定量PCR显示菌株TPEL 13,15,17,18和19覆盖大量的TPEL基因副本，而TPEL 18在用拷贝数标准化时测试的所有CP菌株中的TPEL基因的副本最高16S RRNA基因作为管家基因标记。使用多个口服CP接种的体内NE挑战试验与高蛋白饮食结合，表明TPEL 17是诱导典型的NE病变中最具疾病的致毒性，其次是在商业肉鸡鸡中测试时作为下一个最具毒性的TPEL 19。用TPEL感染17显着降低了生长速率，如在第5天的第5天的相对体重增加百分比所示。毒性NetB（+）TPEL（+）CP株的可用性对于开发CP-Inone NE挑战模型至关重要，该模型可以为了解CP发病机制和抗生素的替代方案提供重要的工具。

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