Desiccation-based microbial sample preservation in remote regions

偏远地区基于干燥的微生物样本保存

• 批准号: 2243038
• 负责人: Stilianos Louca
• 金额: $ 19.05万
• 依托单位: University of Oregon Eugene
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-10-01 至 2025-09-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2243038&HistoricalAwards=false
• 关键词: Desiccation; based; microbial; sample; preservation

An award is made to the University of Oregon to develop and evaluate non-cryogenic methods for microbial sample preservation in the field based on heat-assisted desiccation, a promising approach that presents several practical and economical advantages over existing sample preservation methods. The methods explored will be designed to have low risk and cost, result in low sample weight, and be practical in remote and less developed regions (e.g., requiring only a vehicle's auxiliary power outlet). The insights gained will enable researchers around the world to survey a substantially broader range of locations with reduced logistical challenges. In addition, because dried material is typically safer, lighter and does not require a cold chain, it is also easier to transport and ship between countries, thus facilitating international collaborations. The portable and economical desiccation methods explored here would greatly facilitate surveys of potentially contaminated water bodies and feces during disease outbreaks, as well as the proactive archival of collected material for future analyses, thus contributing to improved disease management and prevention. In addition, desiccated sample material can be archived over prolonged periods of time at room temperatures, and can thus serve as a cheaper alternative to energy-hungry cryopreservation or as a safety backup in the case of freezer failures. The project will also provide research opportunities to one or more undergraduate students.Modern sequencing methods are revolutionizing microbiome research. However, sample preservation in the field and during transport remains a frequent bottleneck that entails substantial logistical challenges, particularly in remote areas or underdeveloped countries. Indeed, the golden standard in sample preservation, flash-freezing at liquid nitrogen temperatures and subsequent storage in a freezer, is often impractical or outright impossible. While alternative non-cryogenic sample preservation methods exist, each of them comes with a set of limitations. Heat-assisted desiccation is a promising simple and economical approach that has so far received little attention. This project will examine multiple alternative protocols of heat-assisted desiccation in a factorial manner. The project will focus on the ability of the explored methods to yield suitable DNA for 16S rRNA marker gene sequencing and metagenomics (two of the most important techniques in microbiome research), after short-term and long-term sample storage at room temperature. The project will thus provide the first systematic evaluation of heat-assisted sample desiccation in terms of DNA yield, DNA integrity, taxonomic biases and suitability for 16S rRNA and genome-resolved metagenomic sequencing. The project would also serve as an important milestone for the future development of commercial miniaturized heat-based dehydrators optimized for biological research in remote regions.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

授予俄勒冈州大学一项奖项，以开发和评估基于热辅助干燥的现场微生物样品保存的非低温方法，这是一种有前途的方法，与现有的样品保存方法相比具有几个实用和经济的优势。所探讨的方法将设计为风险和成本低，样本重量低，并在偏远和欠发达地区（例如，仅需要车辆的辅助电源插座）。所获得的见解将使世界各地的研究人员能够在减少后勤挑战的情况下调查更广泛的地点。此外，由于干燥材料通常更安全，更轻，不需要冷链，因此更容易在国家之间运输和运输，从而促进国际合作。这里探讨的便携式和经济的干燥方法将大大有助于在疾病爆发期间调查可能受污染的水体和粪便，以及对收集的材料进行主动存档以供未来分析，从而有助于改善疾病管理和预防。此外，干燥的样品材料可以在室温下长时间存档，因此可以作为耗能的低温保存的更便宜的替代方案，或者在冷冻机故障的情况下作为安全备份。该项目还将为一名或多名本科生提供研究机会。现代测序方法正在彻底改变微生物组研究。然而，在实地和运输过程中的样本保存仍然是一个经常遇到的瓶颈，带来了巨大的后勤挑战，特别是在偏远地区或不发达国家。事实上，样品保存的黄金标准，即在液氮温度下快速冷冻并随后储存在冰箱中，通常是不切实际的或完全不可能的。虽然存在替代的非低温样品保存方法，但每种方法都有一系列限制。热辅助干燥是一种有前途的简单而经济的方法，迄今为止很少受到关注。本项目将以析因方式研究热辅助干燥的多种替代方案。该项目将重点关注所探索的方法在室温下短期和长期样品储存后产生适合16S rRNA标记基因测序和宏基因组学（微生物组研究中最重要的两项技术）的DNA的能力。因此，该项目将提供第一个系统的评估热辅助样品干燥的DNA产量，DNA完整性，分类偏差和16S rRNA和基因组分辨宏基因组测序的适用性。该项目也将作为一个重要的里程碑，为未来的商业小型化热为基础的制冷机的发展，优化在偏远地区的生物研究。这一奖项反映了NSF的法定使命，并已被认为是值得的支持，通过评估使用基金会的知识价值和更广泛的影响审查标准。