UNS: Persistence of Molecular Markers Used in Fecal Source Tracking: Metagenomic-Based Assessment of Microbial and Human DNA

UNS：粪便来源追踪中使用的分子标记的持久性：基于宏基因组的微生物和人类 DNA 评估

• 批准号: 1511825
• 负责人: Joe Brown
• 金额: $ 33万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1511825&HistoricalAwards=false
• 关键词: UNS; Persistence; Molecular; Markers; Used

1511825BrownWith the relatively new molecular biological tools, the identification of fecal contamination in the environment increasingly makes use of rapid molecular assays to identify DNA sequences specific to fecal indicator bacteria or DNA from host-specific cells such as mitochondrial DNA (mtDNA) shed in stools. Unknowns persist, however, primarily regarding the kinetic relationships between culture-dependent and culture-independent assays, since time-since-contamination information is critical in surveillance as a proxy for fecal pathogen survival and therefore public health risk. The proposed work will elucidate the strengths and limitations of current and novel molecular assays intended to identify fecal contamination in environmental waters.To provide new insights into these issues, the research team will undertake fundamental work on degradation of human mtDNA and human-specific microbial DNA-based targets in parallel with a suite of commonly used culture and qPCR methods for fecal source-tracking applications using laboratory-based mesocosms that simulate well natural surface water and sediments. Mesocosms will be spiked with sources of fecal contamination (healthy and diarrheal stools, primary effluent, isolates) and the fate of the gut microbial community, autochthonous microbial community, and pathogens (when present) will be followed over time (6 months) with advanced metagenomics techniques. Metagenomic approaches can track changes to microbial communities and pathogens with unprecedented accuracy and sensitivity. Hence, the temporal metagenomic datasets will provide an ideal reference to benchmark and compare the efficacy of currently used (fecal indicator bacteria test) and proposed (e.g., human mtDNA) biomarkers for fecal pollution monitoring, revealing comparative kinetic models and potentially new, robust targets for fecal source tracking. Further, the project has the potential to identify promising additional biomarkers for fecal contamination, resulting in improvements in risk assessment tools used by water quality managers. Hence, these studies will likely lead to the development of novel advanced techniques for water quality monitoring as well. Results of this research will be broadly disseminated to the public via a joint workshop with a local partner, Chattahoochee River Keeper, and an interactive exhibit called the Microbe Petting Zoo at the Georgia State Fair.

1511825布朗与相对较新的分子生物学工具，越来越多地利用快速分子测定在环境中鉴定粪便污染，以鉴定粪便指示剂细菌特异性的DNA序列或宿主特异性细胞（例如线粒体DNA（MTDNA）（mtdna））sheed的DNA序列。然而，未知的主要是关于文化依赖性和与文化无关的测定之间的动力学关系的，因为时间 - 污染信息对于监视至关重要，这是粪便病原体生存的代理，因此是公共卫生的风险。拟议的工作将阐明旨在确定环境水域粪便污染的当前和新型分子测定的优势和局限模拟天然地表水和沉积物的中焦。中焦将在粪便污染（健康和腹泻粪便，原发性流出物，分离株）和肠道微生物群落，自我冷思微生物群落和病原体（当下）随着先进的Metagenomics技术的遵循。宏基因组方法可以以前所未有的准确性和敏感性跟踪微生物群落和病原体的变化。因此，时间元基因组数据集将提供基准的理想参考，并比较当前使用的（粪便指标细菌测试）的功效，并提出（例如，人mtDNA）生物标志物对粪便污染监测，揭示了比较的动力学模型以及潜在的新型动力学模型以及对粪便源跟踪的鲁棒目标。此外，该项目有可能确定有希望的粪便污染的其他生物标志物，从而改善了水质经理使用的风险评估工具。因此，这些研究也可能导致开发用于水质监测的新型高级技术。这项研究的结果将通过与当地合作伙伴Chattahoochee River Keeper的联合研讨会广泛传播给公众，以及在佐治亚州博览会上的互动展览，名为Microbe Petting动物园。

项目成果

Toward shotgun metagenomic approaches for microbial source tracking sewage spills based on laboratory mesocosms

• DOI: 10.1016/j.watres.2021.117993
• 发表时间: 2022-02-15
• 期刊: WATER RESEARCH
• 影响因子: 12.8
• 作者: Lindner，Blake G.;Suttner，Brittany;Konstantinidis，Konstantinos T.
• 通讯作者: Konstantinidis，Konstantinos T.