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标准猪舍中细菌生物气溶胶多样性的非培养方法及季节效应评估。

Culture-independent approach of the bacterial bioaerosol diversity in the standard swine confinement buildings, and assessment of the seasonal effect.

作者信息

Nehme Benjamin, Létourneau Valérie, Forster Robert J, Veillette Marc, Duchaine Caroline

机构信息

Centre de Recherche, Institut de Recherche en Cardiologie et en Pneumologie, Hôpital Laval, Québec, QC, Canada.

出版信息

Environ Microbiol. 2008 Mar;10(3):665-75. doi: 10.1111/j.1462-2920.2007.01489.x.

DOI:10.1111/j.1462-2920.2007.01489.x
PMID:18237302
Abstract

The bacterial bioaerosol community of eight swine confinement buildings (SCB) was monitored during two visits in the winter, and one during the summer. To our knowledge, culture-independent approaches and molecular biology tools such as biomass quantification and biodiversity analyses have never been applied to swine building bioaerosol analyses. Total DNA of each sample was extracted and analysed by quantitative real-time polymerase chain reaction, denaturing gradient gel electrophoresis (DGGE) and phylogenetic analysis using primers targeting the bacterial 16S rRNA gene. Even though the total bacterial concentration was higher in winter than in summer, the total bacterial concentration for both seasons was 100 to1000 times higher than the total cultural bacteria. The concentration of bioaerosol was influenced by the temperature indoors, which was regulated with an electronic fan system driving warm air and particles outside of the SCB. Comparison of the DGGE profiles showed the same biodiversity in each SCB during both seasons. The phylogenetic analysis revealed a large number of sequences (93.8%) related to Gram-positive anaerobic bacteria, such as Clostridia, and dominated by the Clostridia cluster I (C. disporicum) and the Clostridia cluster XI (C. glycolycum). The bioaerosol diversity also contained also a low proportion of Bacteroidetes and Lactobacillales-Streptococcales sequences. Analyses of the global community and phylotype diversity showed that the main source of bioaerosols could come from the pig manure slurry.

摘要

在冬季的两次走访以及夏季的一次走访期间,对八座猪舍(SCB)中的细菌生物气溶胶群落进行了监测。据我们所知,诸如生物量定量和生物多样性分析等非培养方法和分子生物学工具从未应用于猪舍生物气溶胶分析。提取每个样本的总DNA,并通过定量实时聚合酶链反应、变性梯度凝胶电泳(DGGE)以及使用靶向细菌16S rRNA基因的引物进行系统发育分析。尽管冬季的总细菌浓度高于夏季,但两个季节的总细菌浓度均比总培养细菌高100至1000倍。生物气溶胶的浓度受室内温度影响,室内温度通过电子风扇系统调节,将热空气和颗粒排出猪舍。DGGE图谱的比较显示,两个季节中每个猪舍的生物多样性相同。系统发育分析揭示了大量与革兰氏阳性厌氧菌相关的序列(93.8%),如梭菌属,且以梭菌属第一簇(双孢梭菌)和梭菌属第十一簇(糖解梭菌)为主。生物气溶胶多样性中也包含低比例的拟杆菌门和乳杆菌目-链球菌目序列。对全球群落和系统型多样性的分析表明,生物气溶胶的主要来源可能是猪粪浆。

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