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Culture-Independent Characterization of Archaeal Biodiversity in Swine Confinement Building Bioaerosols

机译:猪封闭建筑生物气溶胶中古细菌生物多样性的文化独立表征。

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摘要

It was previously demonstrated that microbial communities of pig manure were composed of both bacteria and archaea. Recent studies have shown that bacteria are aerosolized from pig manure, but none have ever focused on the airborne archaeal burden. We sought here to develop and apply molecular ecology approaches to thoroughly characterize airborne archaea from swine confinement buildings (SCBs). Eight swine operations were visited, twice in winter and once during summer. Institute of Occupational Medicine cassettes loaded with 25-mm gelatin filters were used to capture the inhalable microbial biomass. The total genomic DNA was extracted and used as a template for PCR amplification of the archaeal 16S rRNA gene. High concentrations of archaea were found in SCB bioaerosols, being as high as 108 16S rRNA gene copies per cubic meter of air. Construction and sequencing of 16S rRNA gene libraries revealed that all sequences were closely related to methanogenic archaea, such as Methanosphaera stadtmanae (94.7% of the archaeal biodiversity). Archaeal community profiles were compared by 16S rRNA gene denaturing gradient gel electrophoresis. This analysis showed similar fingerprints in each SCB and confirmed the predominance of methanogenic archaea in the bioaerosols. This study sheds new light on the nature of bioaerosols in SCBs and suggests that archaea are also aerosolized from pig manure.
机译:先前已证明,猪粪中的微生物群落由细菌和古细菌组成。最近的研究表明,猪粪中的细菌已被雾化,但从未有过关注空气中古生菌的负担。我们在这里寻求开发和应用分子生态学方法,以彻底表征猪场建筑物(SCB)中的空中古细菌。进行了八次猪场手术,冬季两次,夏季一次。装有25毫米明胶过滤器的职业医学研究所磁带用于捕获可吸入的微生物生物量。提取总的基因组DNA,并将其用作用于PCR扩增古细菌16S rRNA基因的模板。在SCB生物气溶胶中发现高浓度的古细菌,每立方米空气中高达10 8 16S rRNA基因拷贝。 16S rRNA基因文库的构建和测序表明,所有序列均与产甲烷的古生菌密切相关,如产甲烷菌(占甲烷古生物多样性的94.7%)。通过16S rRNA基因变性梯度凝胶电泳比较古细菌群落概况。该分析显示每个SCB中的指纹相似,并证实了甲烷气生古菌在生物气溶胶中的优势。这项研究为SCBs中生物气溶胶的性质提供了新的思路,并表明古细菌也从猪粪中被气雾化。

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