EAGER:COLLABORATIVE RESEARCH: Metagenomic Analysis to ensure water quality and safety

1742869/1742870Huq；Leddy随着水资源短缺的加剧，维持安全的水供应至关重要，包括用于环境排放或再利用的废水。目前，微生物(细菌和病毒)水质是使用标准微生物和生物技术培养方法来测量的，以确定安全的水质。这项研究具有变革性，因为这项研究旨在探索利用新开发的详细和灵敏的元基因组序列分析来了解和鉴定微生物群落，以准确确定水、废水、生物膜和水回用系统中的生物水质。利用拟议的元基因组分析系统，将准确地鉴定微生物，包括细菌、病毒、真菌和寄生虫。此外，还对抗生素耐药性和致病特性进行了鉴定，以现场评估基因和微生物的多样性和丰度。通过评估微生物群落和比较它们的组成，更好地了解微生物群落及其生态，为间接饮用水再利用和直接饮用水再利用的微生物水质评估提供改进的方法。依赖培养的方法和指示性微生物测量已被证明是缓慢和有偏见的，因为它们不能代表所有物种，往往甚至不是深度水和废水处理过程中存在的优势物种。分子技术提供了对微生物群落结构的洞察；然而，这些方法受到严重限制，因为它们针对的是通常不太占优势的单一微生物。因此，通过使用下一代测序进行元基因组分析，可以准确地解析微生物群落及其多样性，从而为环境和公共卫生应用带来显著好处。元基因组全DNA测序包括剪切样本中的所有DNA，然后通过下一代测序进行测序。下一代测序是非歧视性的，识别所有生物体，即使是代表性不足的生物体，同时识别功能和抗性基因、致病性，并提供基于系统发育树的分类结构，所有这些都在一次分析中进行，大约一天内完成。因此，下一代测序提供了更完整的鉴定，对微生物群落及其生态的分辨率更高。这项研究的结果有可能在过程的每个步骤确定水系统中的微生物群落，并将改变理解和设计更好的指标、验证替代品的能力，并定义有利于饮用水和水再利用的可持续微生物群落。长期的社会影响将是至关重要的，因为拟议的项目将为水质和安全供水产生更明确的微生物群落结构，并将展示这一重要的、正在出现的方法对水和水再利用行业的效用。在这项研究的过程中，博士生将获得生物信息学和水再利用领域的基础和应用知识，该领域包括用于水再利用的高级处理技术。学生实习生将有机会学习现场采样技术。这项研究将为实现显著的更广泛的环境影响奠定良好的基础。

1742869/ 1742870Huq; LeddyWith increased water scarcity, it is critical that safe water supplies be maintained, including wastewater for environmental discharge or reuse. Currently microbial (bacterial and viral) water quality is measured using standard microbiological and biotechnological culture methods to establish safe water quality. This proposed EAGER is transformative as the study aims to explore the use of a newly developed detailed and sensitive metagenomic sequence analysis to understand and identify microbial communities for accurate determination of biological water quality in water, wastewater, biofilms, and water reuse systems.With the proposed metagenomic analysis system, microorganisms, including bacteria, viruses, fungi, and parasites, will be accurately identified to species and strains. Additionally, antibiotic resistance and pathogenicity properties are also identified to evaluate diversity and abundance of the genes and microorganisms in-situ. By assessing the microbial communities and comparing their composition, a better understanding of the microbial communities and their ecology is developed to provide improved methods for assessing microbial water quality for indirect potable reuse and direct potable reuse. Culture-dependent methods and indicator microorganism measurements have proven to be slow and biased because they do not represent all species, often not even the dominant species present during advanced water and wastewater treatment. Molecular techniques have provided insights into microbial community structure; however, these methods are seriously limited because they target single microorganisms that are often the less dominant species. Therefore, with metagenomic analysis using next generation sequencing, accurate resolution of the microbial communities and their diversity is achieved, resulting in significant benefits for environmental and public health applications. Metagenomic whole DNA sequencing involves shearing all DNA in a sample that is then sequenced by next generation sequencing. Next generation sequencing is non-discriminatory, identifying all organisms, even the underrepresented, while identifying functional and resistant genes, pathogenicity, and providing taxonomic structure based on phylogenetic trees, all in a single analysis and within approximately one day. Therefore, next generation sequencing provides a more complete identification, with greater resolution of the microbial community and its ecology. The results from this study have the potential of defining microbial communities in water systems at each step of the process and will transform the ability to understand and design better indicators, validate surrogates, and also define sustainable microbial communities beneficial for drinking water and water reuse. The long term societal impact will be paramount as the proposed project will yield a more defined microbial community structure for water quality and safe water supplies and will demonstrate the utility of this important, emerging approach for the water and water reuse industry. During the course of this study, a doctoral student will gain fundamental and applied knowledge in the area of bioinformatics and water reuse that encompasses advanced treatment technologies that are employed in water reuse. Student interns will have the opportunity to learn field sampling techniques. This study will establish a good foundation to achieve notable broader impacts for the environment.