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Prevalence of shiga toxin-producing escherichia coli and associated virulence genes in feces of commercial feedlot cattle

机译:商业饲养场牛粪便中产生志贺毒素的大肠杆菌及其相关毒力基因的流行

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摘要

The objective of this study was to determine the prevalence of Shiga toxin-producing Escherichia coli (STEC) serogroups and associated virulence genes in feces of commercial feedlot cattle. During March to May 2011, fecal samples were collected from individual cattle (n=960) in 10 cohorts (cattle subpopulations within a feedlot) comprising 17,148 total steers that originated from 48 backgrounding operations in six U.S. states. Fecal samples were enriched in E. coli broth and subjected to two detection protocols: (1) an 11-gene multiplex polymerase chain reaction (PCR) that identifies seven O serogroups (O26, O45, O103, O111, O121, O145, and O157) and four virulence genes (stx1, stx2, eae, and ehxA) applied to extracted total DNA ("direct PCR"); and (2) cultural procedures that involve immunomagnetic separation (IMS) with O26, O103, and O111 beads, plating on a nondifferential MacConkey agar, followed by the multiplex PCR of pooled colonies ("culture-based method"). Generalized linear mixed models were used to adjust prevalence estimates for clustering. Based on direct PCR detection, O157 (49.9%) was the most prevalent O serogroup followed by O26 (20.3%), O103 (11.8%), O121 (10.7%), O45 (10.4%), O145 (2.8%), and O111 (0.8%). Cumulative adjusted prevalence estimates were 22.3, 24.6, and 0.01% for O26, O103, and O111 serogroups, respectively, based on culture-based methods. However, prevalence varied significantly by cohort (p-values<0.05) for O26, O121, and O157 based on direct PCR, and for O26, O103, and O111 serogroups based on culture-based methods. Results of this study indicate that all seven STEC serogroups were identified in feedlot cattle feces, with O157, O26, and O103 being the most prevalent serogroups. A substantial proportion of serogroup-positive samples did not harbor Shiga toxin genes; thus, additional elucidation of the potential human health risk is required. Further evaluation of diagnostic methods for non-O157 STEC is needed given their impact on prevalence estimation.
机译:这项研究的目的是确定商业饲育牛粪便中产生志贺毒素的大肠杆菌(STEC)血清群和相关毒力基因的患病率。在2011年3月至2011年5月期间,从10个队列(育肥场内的牛亚群)的每头牛(n = 960)中收集了粪便样本,这些粪便样本总共有17,148头,来自美国六个州的48个背景调查。粪便样品富含大肠杆菌肉汤,并接受两种检测方案:(1)11基因多重聚合酶链反应(PCR),可识别七个O血清群(O26,O45,O103,O111,O121,O145和O157 )和应用于提取的总DNA的四个毒力基因(stx1,stx2,eae和ehxA)(“直接PCR”); (2)培养程序,包括用O26,O103和O111磁珠进行免疫磁分离(IMS),在无差异的MacConkey琼脂上铺板,然后对合并的菌落进行多重PCR(“基于培养物的方法”)。广义线性混合模型用于调整患病率估计值。根据直接PCR检测,O157(49.9%)是最普遍的O血清群,其次是O26(20.3%),O103(11.8%),O121(10.7%),O45(10.4%),O145(2.8%)和O111(0.8%)。根据基于文化的方法,O26,O103和O111血清群的调整后患病率累计估计分别为22.3、24.6和0.01%。但是,基于直接PCR的O26,O121和O157血清群和基于培养法的O26,O103和O111血清群的队列患病率差异显着(p值<0.05)。这项研究的结果表明,在饲养场牛粪中鉴定出所有七个STEC血清群,其中O157,O26和O103是最普遍的血清群。血清群阳性样本中有很大一部分没有携带志贺毒素基因。因此,需要进一步阐明潜在的人类健康风险。考虑到非O157 STEC诊断方法对患病率估计的影响,需要进一步评估。

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