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使用 MinION 纳米孔测序技术检测正常牛奶中的微生物菌群。

Examination of the microbiota of normal cow milk using MinION nanopore sequencing.

机构信息

School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa 252-5201, Japan.

Azabu University Mastitis Research Center, Sagamihara, Kanagawa 252-5201, Japan.

出版信息

J Vet Med Sci. 2021 Oct 31;83(11):1620-1627. doi: 10.1292/jvms.21-0353. Epub 2021 Sep 15.

Abstract

The aim of this study was to evaluate the microbiota of normal milk in dairy cows and their relationship with host factors, such as the age of the cow (Age), somatic cell counts in milk (SCCs), and days in milk (DIM). We investigated 48 milk samples from 22 cows with no systemic or local clinical signs using MinION nanopore sequencing for a 16S rRNA gene amplicon. Bacterial richness was positively correlated with the DIM (P=0.043), and both the Shannon-Wiener Index and Simpson's Index, which are metrics of alpha-diversity, were also significantly positively correlated with the SCC (P<0.001). The composition ratios of both Actinobacteria at the phylum level and Kocuria spp. at the genus level in the milk microbiota were significantly correlated with the SCC (P<0.001 and P<0.001, respectively). In the beta-diversity test, the one-way analysis of similarities test showed a significant difference (P=0.0051) between the low- and high-SCC groups. This study clarified that the composition of the normal milk microbiota in this herd was related to the SCC. It also raised the possibility of variations in bacterial genera in the normal milk microbiota between the low- and high-SCC groups. However, to clarify the actual condition of the milk microbiota and to elucidate the relationship with the SCC, it is necessary to perform further analyses taking into account not only the relative abundance, but also the absolute abundance of microbes.

摘要

本研究旨在评估奶牛正常乳中的微生物群及其与宿主因素(如奶牛年龄(Age)、乳中体细胞计数(SCCs)和泌乳天数(DIM))的关系。我们使用 MinION 纳米孔测序对 22 头无全身性或局部临床症状的奶牛的 48 份乳样进行了研究,共获得 16S rRNA 基因扩增子。细菌丰富度与 DIM 呈正相关(P=0.043),Shannon-Wiener 指数和 Simpson 指数这两个α多样性指标也与 SCC 呈显著正相关(P<0.001)。乳微生物群中厚壁菌门水平的放线菌和乳球菌属水平的科库利亚菌的组成比例与 SCC 显著相关(P<0.001 和 P<0.001)。在β多样性检验中,相似性的单向分析检验(one-way analysis of similarities test)表明低 SCC 组和高 SCC 组之间存在显著差异(P=0.0051)。本研究表明,该牛群的正常乳微生物群组成与 SCC 有关。这也提示了在低 SCC 组和高 SCC 组之间,正常乳微生物群中的细菌属可能存在差异。然而,为了阐明乳微生物群的实际状况,并阐明与 SCC 的关系,有必要在考虑微生物相对丰度的基础上,进一步考虑绝对丰度进行分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bab7/8636880/c670f882a19c/jvms-83-1620-g001.jpg

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