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牛流产材料中细菌微生物群的扩增子测序。

Amplicon sequencing of bacterial microbiota in abortion material from cattle.

作者信息

Vidal Sara, Kegler Kristel, Posthaus Horst, Perreten Vincent, Rodriguez-Campos Sabrina

机构信息

Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Bern, Laenggassstrasse 122, 3012, Bern, Switzerland.

Graduate School for Cellular and Biomedical Sciences, Theodor Kocher Institute, University of Bern, Freiestrasse 1, 3001, Bern, Switzerland.

出版信息

Vet Res. 2017 Oct 10;48(1):64. doi: 10.1186/s13567-017-0470-1.

Abstract

Abortions in cattle have a significant economic impact on animal husbandry and require prompt diagnosis for surveillance of epizootic infectious agents. Since most abortions are not epizootic but sporadic with often undetected etiologies, this study examined the bacterial community present in the placenta (PL, n = 32) and fetal abomasal content (AC, n = 49) in 64 cases of bovine abortion by next generation sequencing (NGS) of the 16S rRNA gene. The PL and AC from three fetuses of dams that died from non-infectious reasons were included as controls. All samples were analyzed by bacterial culture, and 17 were examined by histopathology. We observed 922 OTUs overall and 267 taxa at the genus level. No detectable bacterial DNA was present in the control samples. The microbial profiles of the PL and AC differed significantly, both in their composition (PERMANOVA), species richness and Chao-1 (Mann-Whitney test). In both organs, Pseudomonas was the most abundant genus. The combination of NGS and culture identified opportunistic pathogens of interest in placentas with lesions, such as Vibrio metschnikovii, Streptococcus uberis, Lactococcus lactis and Escherichia coli. In placentas with lesions where culturing was unsuccessful, Pseudomonas and unidentified Aeromonadaceae were identified by NGS displaying high number of reads. Three cases with multiple possible etiologies and placentas presenting lesions were detected by NGS. Amplicon sequencing has the potential to uncover unknown etiological agents. These new insights on cattle abortion extend our focus to previously understudied opportunistic abortive bacteria.

摘要

牛流产对畜牧业有重大经济影响,需要及时诊断以监测动物流行病病原体。由于大多数流产并非动物流行病而是散发的,病因往往未被发现,本研究通过对16S rRNA基因进行下一代测序(NGS),检查了64例牛流产病例中胎盘(PL,n = 32)和胎儿皱胃内容物(AC,n = 49)中的细菌群落。将死于非感染性原因的母畜所产三个胎儿的PL和AC作为对照。所有样本均进行细菌培养分析,17个样本进行组织病理学检查。我们总共观察到922个操作分类单元(OTU)和267个属水平的分类群。对照样本中未检测到细菌DNA。PL和AC的微生物谱在组成(PERMANOVA)、物种丰富度和Chao-1指数(曼-惠特尼检验)方面均有显著差异。在两个器官中,假单胞菌属是最丰富的属。NGS和培养相结合,在有病变的胎盘中鉴定出了感兴趣的机会性病原体,如梅氏弧菌、乳房链球菌、乳酸乳球菌和大肠杆菌。在培养未成功的有病变的胎盘中,通过NGS鉴定出假单胞菌属和未鉴定的气单胞菌科,其读数很高。通过NGS检测到三例有多种可能病因且胎盘有病变的病例。扩增子测序有潜力发现未知病原体。这些关于牛流产的新见解将我们的关注点扩展到了以前研究较少的机会性流产细菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48da/5633877/02dcb5e5e8ea/13567_2017_470_Fig3_HTML.jpg

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