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Can air microbiota be a novel marker for public health? A sampling model and preliminary data from different environments

机译:空气微生物群能成为公共卫生的新标志吗?来自不同环境的采样模型和初步数据

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Next-generation sequencing (NGS) and advances in microbiota knowledge are opening new perspectives for environmental microbiology and public health. Massive parallel sequencing of bacterial DNA is providing new insights to unravel microbial dissemination in different matrices, including air. Here, we have assessed the air biodiversity in several indoor environments by NGS-based metagenomics and compared results with respect to classical approaches. Environmental sampling was achieved in three exemplificative indoor locations: offices, recreational facilities and public restrooms. Sampling was performed by active (surface air system and high-volume aerosol collection system) and passive (index of microbial air contamination and novel tissue-based passive sampling) techniques. Collected samples were tested by traditional microbiology and DNA-based methods. Biodiversity indices were computed using EstimateS software. Statistical analysis by SPSS and R was applied to determine the variability of bacteria groups after comparing sampling and analysis protocols. In each point (n = 6), air was sampled in duplicate by four methods and tested in parallel by culture-based and DNA-based approaches. A total of over 30 +/- 10 isolates for each point were identified at species level. A total of 3,207,415 sequence reads were generated by NGS analysis, leading to the identification of over 170 OTUs. NGS analyses of air biodiversity were more informative when using active sampling methods (0.729, p < 0.05) and less by passive methods (- 0.897, p < 0.05). NGS allowed a comprehensive and reproducible evaluation of air microbiota and the definition of microbial signatures. Air can be assayed by NGS using different strategies, representing a promising tool for evaluating microbial load and biodiversity. Although NGS requires sophisticated equipment, bioinformatics tools and trained personnel, we propose a model for the evaluation of indoor air quality in occupational and public health settings.
机译:下一代测序(NGS)和微生物群落知识的进步为环境微生物学和公共卫生开辟了新的前景。细菌DNA的大规模并行测序为揭示微生物在不同基质(包括空气)中的传播提供了新的见识。在这里,我们通过基于NGS的宏基因组学评估了几种室内环境中的空气生物多样性,并比较了传统方法的结果。在三个示例性室内位置进行了环境采样:办公室,娱乐设施和公共洗手间。通过主动(地面空气系统和大量气溶胶收集系统)和被动(微生物空气污染指数以及基于新型组织的被动采样)技术进行采样。收集的样品通过传统的微生物学和基于DNA的方法进行测试。使用评估软件计算生物多样性指数。在比较采样和分析方案后,通过SPSS和R进行统计分析,以确定细菌组的变异性。在每个点(n = 6)中,通过四种方法一式两份采样空气,并通过基于培养物和基于DNA的方法并行测试。在物种水平上,每个点总共鉴定出30 +/- 10个分离株。通过NGS分析,总共产生了3,207,415个序列读数,从而鉴定出170多个OTU。使用主动采样方法(0.729,p <0.05)时,对空气生物多样性的NGS分析提供的信息更多,而通过被动方法(-0.897,p <0.05)则较少。 NGS可以对空气微生物群和微生物特征进行全面,可重复的评估。 NGS可以使用不同的策略来分析空气,这是评估微生物负荷和生物多样性的有前途的工具。尽管NGS需要复杂的设备,生物信息学工具和训练有素的人员,但我们提出了一种用于在职业和公共卫生场所评估室内空气质量的模型。

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