Microbiota in Exhaled Breath Condensate and the Lung.
Microbiota in Exhaled Breath Condensate and the Lung.
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DOI：
10.1128/aem.00515-17
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发表时间：
2017-06-15
影响因子：
4.4
通讯作者：
McLachlan G
中科院分区：
文献类型：
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作者：
Glendinning L;Wright S;Tennant P;Gill AC;Collie D;McLachlan G
肺部微生物区系通常使用相对侵入性的支气管镜检查程序进行采样。呼气冷凝液(EBC)采集可能为肺微生物区系采样提供了一种侵入性较小的替代方案。我们比较了保护性标本刷法(PSB)和呼气冷凝液收集法(PSB)提取的肺微生物区系样本。我们还试图评估雾化抗生素治疗是否会影响肺部微生物区系，以及这种变化是否可以在EBC中检测到。收集清醒绵羊6只，麻醉绵羊6只，在机械通气时采集EBC。在后一次EBC采集之后，从每个绵羊肺内的不同部位采集了PSB样本。第二天，每只羊都被雾化吸入氨基甲酸酯钠。雾化两天后，再次采集EBC和PSB样本。用16S rRNA基因定量聚合酶链式反应检测细菌DNA。用聚合酶链式反应扩增16S rRNA基因的V2-V3区，并用Illumina MiSeq测序。用MOTHUR进行质量控制和操作分类单元(OTU)聚类。EBC样品的细菌DNA含量明显低于PSB样品。麻醉动物的EBC样本与PSB样本的细菌群落组成不同地聚在一起，37个细菌OTU被鉴定为两种样本之间的差异丰富。尽管在支气管肺泡灌洗液中只检测到低浓度的粘菌素，但在氨甲酸酯钠治疗前后，PSB样本的细菌组成有所不同。我们的发现表明，EBC样品和PSB样品中的微生物区系不是相同的。对肺部微生物区系进行重要采样通常需要进行支气管镜检查程序，包括对人类参与者进行医院访问和使用训练有素的工作人员。参与这类研究的不便和感觉上的不适可能会阻止健康的志愿者，对于晚期肺部疾病患者来说可能不是一个安全的选择。这项研究旨在评估一种侵入性较小的肺微生物区系样本收集方法，方法是比较通过受保护的样本刷子(PSB)和呼气冷凝液(EBC)收集的样本。我们发现，EBC样品中的细菌DNA少于PSB样品中的细菌DNA，并且两种样品中的细菌群落存在差异。我们的结论是，虽然EBC和PSB样品不能产生同等的微生物区系样品，但EBC微生物区系的研究可能仍然有意义。
The lung microbiota is commonly sampled using relatively invasive bronchoscopic procedures. Exhaled breath condensate (EBC) collection potentially offers a less invasive alternative for lung microbiota sampling. We compared lung microbiota samples retrieved by protected specimen brushings (PSB) and exhaled breath condensate collection. We also sought to assess whether aerosolized antibiotic treatment would influence the lung microbiota and whether this change could be detected in EBC. EBC was collected from 6 conscious sheep and then from the same anesthetized sheep during mechanical ventilation. Following the latter EBC collection, PSB samples were collected from separate sites within each sheep lung. On the subsequent day, each sheep was then treated with nebulized colistimethate sodium. Two days after nebulization, EBC and PSB samples were again collected. Bacterial DNA was quantified using 16S rRNA gene quantitative PCR. The V2-V3 region of the 16S rRNA gene was amplified by PCR and sequenced using Illumina MiSeq. Quality control and operational taxonomic unit (OTU) clustering were performed with mothur. The EBC samples contained significantly less bacterial DNA than the PSB samples. The EBC samples from anesthetized animals clustered separately by their bacterial community compositions in comparison to the PSB samples, and 37 bacterial OTUs were identified as differentially abundant between the two sample types. Despite only low concentrations of colistin being detected in bronchoalveolar lavage fluid, PSB samples were found to differ by their bacterial compositions before and after colistimethate sodium treatment. Our findings indicate that microbiota in EBC samples and PSB samples are not equivalent. IMPORTANCE Sampling of the lung microbiota usually necessitates performing bronchoscopic procedures that involve a hospital visit for human participants and the use of trained staff. The inconvenience and perceived discomfort of participating in this kind of research may deter healthy volunteers and may not be a safe option for patients with advanced lung disease. This study set out to evaluate a less invasive method for collecting lung microbiota samples by comparing samples taken via protected specimen brushings (PSB) to those taken via exhaled breath condensate (EBC) collection. We found that there was less bacterial DNA in EBC samples compared with that in PSB samples and that there were differences between the bacterial communities in the two sample types. We conclude that while EBC and PSB samples do not produce equivalent microbiota samples, the study of the EBC microbiota may still be of interest.