Effects of Vacuum-Heat-Assisted Sample Desiccation on Microbiome Surveys.

IF 5.5 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Stilianos Louca, Claire E Mullin
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引用次数: 0

Abstract

Sample preservation in the field and during transport can be a logistical challenge for microbiome surveys, particularly in remote areas. Sample desiccation eliminates the need for complicated cold chains and dangerous preservatives. However, the effects of desiccation on modern microbiome workflows such as gene-centric metagenomic profiling and metagenome-assembled genome (MAG) binning, remain poorly understood. In addition, most common desiccation tools such as lyophilisation cannot easily be deployed in the field. Here, we describe a proof-of-principle sample desiccator using vacuum and heat, specifically built for deployment in the field and exhibiting low power consumption and cost. We then test the effects of vacuum-heat-assisted sample desiccation followed by storage at room temperature, in comparison to conventional freezing, on multiple soil and animal faecal samples, via metagenomic and 16S rRNA amplicon sequencing. We consider multiple metrics related to the success of DNA extraction, sequencing, contig assembly, OTU clustering, gene annotation and MAG construction, as well as effects on inferred microbial community composition. We find that the impact of drying on considered success metrics was almost always either minor, non-significant or positive. For a subset of source materials we observed moderate but statistically significant differences in terms of inferred microbial taxonomic and genetic composition. We conclude that vacuum- and heat-assisted desiccation can be a useful, practical and cost-effective tool for microbiome field surveys, when a high consistency with frozen samples is not required.

真空-热辅助样品干燥对微生物组调查的影响。
野外和运输过程中的样品保存可能是微生物组调查的后勤挑战,特别是在偏远地区。样品干燥消除了复杂的冷链和危险的防腐剂的需要。然而,干燥对现代微生物组工作流程的影响,如以基因为中心的宏基因组分析和宏基因组组装基因组(MAG)分类,仍然知之甚少。此外,大多数常见的干燥工具,如冻干,不容易在现场部署。在这里,我们描述了一种原理验证的样品干燥器,使用真空和热量,专门用于现场部署,具有低功耗和低成本。然后,我们通过宏基因组和16S rRNA扩增子测序,测试了真空热辅助样品干燥后在室温下储存的效果,与传统冷冻相比,对多种土壤和动物粪便样品进行了测试。我们考虑了与DNA提取、测序、序列组装、OTU聚类、基因注释和MAG构建的成功相关的多个指标,以及对推断微生物群落组成的影响。我们发现干燥对考虑的成功度量的影响几乎总是次要的、不显著的或积极的。对于源材料的一个子集,我们在推断的微生物分类和遗传组成方面观察到适度但统计上显着的差异。我们得出的结论是,当不需要与冷冻样品高度一致时,真空和热辅助干燥可以成为微生物组现场调查的有用,实用和具有成本效益的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Molecular Ecology Resources
Molecular Ecology Resources 生物-进化生物学
CiteScore
15.60
自引率
5.20%
发文量
170
审稿时长
3 months
期刊介绍: Molecular Ecology Resources promotes the creation of comprehensive resources for the scientific community, encompassing computer programs, statistical and molecular advancements, and a diverse array of molecular tools. Serving as a conduit for disseminating these resources, the journal targets a broad audience of researchers in the fields of evolution, ecology, and conservation. Articles in Molecular Ecology Resources are crafted to support investigations tackling significant questions within these disciplines. In addition to original resource articles, Molecular Ecology Resources features Reviews, Opinions, and Comments relevant to the field. The journal also periodically releases Special Issues focusing on resource development within specific areas.
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