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Characterization of fungi in office dust: comparing results of microbial secondary metabolites fungal ITS region sequencing viable culture and other microbial indices

机译:办公室灰尘中真菌的特征:比较微生物次生代谢产物真菌ITS区域测序可行培养和其他微生物指标的结果

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摘要

Recent developments in molecular and chemical methods have enabled the analysis of fungal DNA and secondary metabolites, often produced during fungal growth, in environmental samples. We compared three fungal analytical methods by analyzing floor dust samples collected from an office building for fungi using viable culture, internal transcribed spacer (ITS) sequencing, and secondary metabolites using liquid chromatography-tandem mass spectrometry. Of the 32 metabolites identified, 29 had a potential link to fungi with levels ranging from 0.04 (minimum for alternariol monomethylether) to 5,700 ng/g (maximum for neoechinulin A). The number of fungal metabolites quantified per sample ranged from eight to sixteen (average=13/sample). We identified 216 fungal operational taxonomic units (OTUs) with the number per sample ranging from six to twenty-nine (average=18/sample). We identified 37 fungal species using culture and the number per sample ranged from two to thirteen (average=eight/sample). Agreement in identification between ITS sequencing and culturing was weak (kappa=−0.12–0.27). The number of cultured fungal species poorly correlated with OTUs, which did not correlate with the number of metabolites. These suggest that using multiple measurement methods may provide an improved understanding of fungal exposures in indoor environments and that secondary metabolites may be considered as an additional source of exposure.
机译:分子和化学方法的最新发展使得能够对环境样品中的真菌DNA和次级代谢产物进行分析,这些代谢产物通常是在真菌生长过程中产生的。我们比较了三种真菌分析方法,即通过使用可行的培养,内部转录间隔物(ITS)测序和使用液相色谱-串联质谱的次级代谢产物分析从办公楼收集的地板灰尘样品中的真菌。在鉴定出的32种代谢产物中,有29种与真菌有潜在的联系,其水平范围为0.04(交替糖单甲醚的最小值)至5700 ng / g(新棘球蛋白A的最大值)。每个样品中定量的真菌代谢物的数量为8到16(平均值= 13 /样品)。我们确定了216个真菌操作分类单位(OTU),每个样品的数量范围为6到29(平均值= 18 /样品)。我们使用培养方法鉴定了37种真菌,每个样品的数量在2到13个之间(平均值= 8 /样品)。 ITS测序与培养之间的鉴定一致性较弱(kappa = -0.12-0.27)。培养的真菌种类的数量与OTU的相关性很弱,而OTU与代谢物的数量没有相关性。这些表明,使用多种测量方法可以更好地了解室内环境中的真菌暴露,并且次生代谢产物可以被视为额外的暴露来源。

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