Variability of the Sheep Lung Microbiota.
Variability of the Sheep Lung Microbiota.
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DOI：
10.1128/aem.00540-16
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发表时间：
2016-06-01
影响因子：
4.4
通讯作者：
McLachlan G
中科院分区：
文献类型：
--
作者：
Glendinning L;Wright S;Pollock J;Tennant P;Collie D;McLachlan G
测序技术最近促进了健康和疾病期间肺部细菌群落的表征。然而，目前缺乏关于这些健康数据在受试者之间和受试者内部的可变性的信息。本研究试图用健康的成年羊作为我们的模型系统来检验这种变异性。在6只成年羊（年龄，20月龄）的3个不同空间段支气管上采集了受保护的标本刷样（第0天、第1个月和第3个月）。为了进一步探索微生物群的空间变异性，从另一只羊身上采集了更广泛的刷牙样本（n = 16）和咽拭子。利用Illumina MiSeq对细菌16S rRNA基因的V2和V3高变区进行扩增和测序。使用母亲软件包分析DNA序列。定量PCR测定细菌总DNA。一些羊的肺部在不同的采样点含有明显不同的细菌群落，而在其他羊的肺部，气道微生物群似乎相似。在我们的空间变异性研究中，我们观察到与采集样本的肺内深度相关的聚类。肺深是指离声门的距离增加，向尾侧方向发展。我们认为，宿主和当地因素都对绵羊肺微生物群的组成有影响。直到最近，人们还认为肺部是一个无菌的环境，只有在疾病期间微生物才会定植。然而，最近使用测序技术的研究发现，在健康期间，肺部存在一小群细菌，称为“肺部微生物群”。在这项研究中，我们描述了健康绵羊肺微生物群的变异性。羊不仅是经济上重要的动物，而且经常被用作人类呼吸系统疾病的大型动物模型。我们得出的结论是，虽然宿主的影响确实在决定定植在气道上的微生物类型方面发挥了作用，但很明显，当地因素在这方面也起着重要作用。了解这些因素的性质和影响将是了解肺微生物群变异性和功能相关性的关键。
Sequencing technologies have recently facilitated the characterization of bacterial communities present in lungs during health and disease. However, there is currently a dearth of information concerning the variability of such data in health both between and within subjects. This study seeks to examine such variability using healthy adult sheep as our model system. Protected specimen brush samples were collected from three spatially disparate segmental bronchi of six adult sheep (age, 20 months) on three occasions (day 0, 1 month, and 3 months). To further explore the spatial variability of the microbiotas, more-extensive brushing samples (n = 16) and a throat swab were taken from a separate sheep. The V2 and V3 hypervariable regions of the bacterial 16S rRNA genes were amplified and sequenced via Illumina MiSeq. DNA sequences were analyzed using the mothur software package. Quantitative PCR was performed to quantify total bacterial DNA. Some sheep lungs contained dramatically different bacterial communities at different sampling sites, whereas in others, airway microbiotas appeared similar across the lung. In our spatial variability study, we observed clustering related to the depth within the lung from which samples were taken. Lung depth refers to increasing distance from the glottis, progressing in a caudal direction. We conclude that both host influence and local factors have impacts on the composition of the sheep lung microbiota. IMPORTANCE Until recently, it was assumed that the lungs were a sterile environment which was colonized by microbes only during disease. However, recent studies using sequencing technologies have found that there is a small population of bacteria which exists in the lung during health, referred to as the “lung microbiota.” In this study, we characterize the variability of the lung microbiotas of healthy sheep. Sheep not only are economically important animals but also are often used as large animal models of human respiratory disease. We conclude that, while host influence does play a role in dictating the types of microbes which colonize the airways, it is clear that local factors also play an important role in this regard. Understanding the nature and influence of these factors will be key to understanding the variability in, and functional relevance of, the lung microbiota.