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中国 2 家大型奶牛场分离的肺炎克雷伯菌的毒力特征。

Virulence profiles of Klebsiella pneumoniae isolated from 2 large dairy farms in China.

机构信息

Department of Clinical Veterinary Medicine, College of Veterinary Medicine, China Agricultural University, Yuanmingyuan West Road, Beijing 100193, China.

Department of Population Medicine, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada, N1G 2W1.

出版信息

J Dairy Sci. 2021 Aug;104(8):9027-9036. doi: 10.3168/jds.2020-20042. Epub 2021 May 10.

Abstract

We recently reported on the diversity of Klebsiella pneumoniae isolated from dairy herds in China. In our previous work, isolates from subclinical mastitis (SCM) had lower indices of diversity when compared with bacteria from other sources, possibly due to a contagious-like spread of udder adapted strains. Here we explored the virulence profile and capsular types of K. pneumoniae isolated from different sources on 2 dairy farms in China. Our overarching goal was to gain insights on the role of virulence genes toward the severity of mastitis caused by K. pneumoniae. A total of 1,484 samples were collected from clinical mastitis (CM; n = 355), SCM (n = 561), bulk tank milk (BTM; n = 130), and environmental and extramammary (EE) sites (n = 438). From those, 431 K. pneumoniae isolates were obtained, including 129, 77, 66, and 159 isolates from CM, SCM, BTM, and EE samples, respectively. Polymerase chain reactions were used to determine the capsular types and to detect potential virulence genes in all isolates. No significant farm effects were observed when comparing the distribution of most virulence genes in K. pneumoniae isolated from each source. K57 was the most prevalent capsular type in K. pneumoniae from all sources, but with increased detection rate in isolates from CM. entB, kfu, fimH1, mrkD, and β-d-lacZ were frequently detected in K. pneumoniae from all sources. β-d-lacZ, entB, and ituA were more prevalent in isolates from CM, whereas kfu, allS, and nif were more frequently detected in isolates from SCM. ybtS, aerobactin, and rpmA had increased prevalence in K. pneumoniae from BTM when compared with bacteria from other sources. No association was detected between virulence genes and the severity of CM. K57 and the nif gene had the highest discriminatory power to classify isolates from CM and SCM, respectively. Based on our findings, it is likely that K57 is the dominant capsular type in K. pneumoniae causing CM in large Chinese dairy herds. Likewise, we demonstrated that β-d-lacZ is disseminated in K. pneumoniae isolated from large Chinese dairy farms, irrespectively of the source of bacteria. Our results also suggest a low contribution of the virulence profile of K. pneumoniae toward CM severity. Finally, the role of nif in increasing the adaptability to the udder and promoting a contagious-like spread of K. pneumoniae warrants further investigation.

摘要

我们最近报道了中国奶牛场分离的肺炎克雷伯菌的多样性。在我们之前的工作中,与来自其他来源的细菌相比,来自隐性乳腺炎(SCM)的分离株的多样性指数较低,这可能是由于与传染性相关的乳区适应株的传播。在这里,我们研究了在中国的两个奶牛场不同来源的肺炎克雷伯菌的毒力谱和荚膜型。我们的总体目标是了解毒力基因在肺炎克雷伯菌引起的乳腺炎严重程度中的作用。从临床乳腺炎(CM;n=355)、隐性乳腺炎(SCM;n=561)、奶罐奶(BTM;n=130)和环境及非乳腺(EE)部位(n=438)共采集了 1484 个样本。从这些样本中,我们获得了 431 株肺炎克雷伯菌分离株,包括分别来自 CM、SCM、BTM 和 EE 样本的 129、77、66 和 159 株分离株。聚合酶链反应用于确定荚膜型,并检测所有分离株中潜在的毒力基因。当比较从每个来源分离的肺炎克雷伯菌中大多数毒力基因的分布时,没有观察到显著的农场效应。K57 是所有来源的肺炎克雷伯菌中最常见的荚膜型,但在 CM 分离株中的检测率增加。entB、kfu、fimH1、mrkD 和β-d-lacZ 经常在所有来源的肺炎克雷伯菌中检测到。β-d-lacZ、entB 和 ituA 在 CM 分离株中更为常见,而 kfu、allS 和 nif 在 SCM 分离株中更为常见。ybtS、aerobactin 和 rpmA 在 BTM 分离株中的检出率高于其他来源的细菌。未检测到毒力基因与 CM 严重程度之间存在相关性。K57 和 nif 基因对 CM 和 SCM 分离株的分类具有最高的鉴别力。基于我们的发现,K57 很可能是中国大型奶牛场引起 CM 的主要荚膜型。同样,我们证明了β-d-lacZ 在中国大型奶牛场分离的肺炎克雷伯菌中传播,而与细菌来源无关。我们的结果还表明,肺炎克雷伯菌的毒力谱对 CM 的严重程度影响不大。最后,nif 基因在提高对乳区的适应性和促进肺炎克雷伯菌的传染性传播方面的作用值得进一步研究。

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