RII Track-4: NSF: Insights into Pathogenicity of Chlorine-Stressed Bacteria Using Combined In Vitro Assays and Metatranscriptomics

水中的细菌病原体对全球公共卫生构成重大威胁，特别是对免疫系统较弱的个人。目前，氯化是最广泛使用的水消毒工艺，可以灭活水中的微生物，以确保公众和环境健康。虽然氯化后水中的大多数细菌都可以被有效杀死，但有些细菌可能会因对消毒剂的抵抗力而存活。目前还没有一项全面的研究来调查存活细菌感染宿主并引起疾病的能力。该研究基础设施改进RII Track-4奖学金项目将探索水消毒对细菌致病性的影响，这将有助于克服目前关于水处理后微生物风险评估的知识差距。这一目标将通过使用先进的细胞工具和下一代测序技术来实现，这些技术是在美国环境保护署的生物测量分支开发的。从该项目中获得的基本知识有望为利益攸关方和决策者提供有价值的信息，以重新评估当前的水处理过程并减少健康风险。这项研究还将提供培训机会，为代表性不足的学生在新墨西哥州州立大学，这是一个西班牙裔服务Institution.This研究基础设施改善轨道-4 EPSCoR研究员（RII轨道-4）的建议将提供奖学金，以助理教授和培训研究生在新墨西哥州州立大学。这项工作将与俄亥俄州辛辛那提的美国环境保护局流域生态系统特征描述司生物测量分支的研究人员合作进行。了解消毒后存活的细菌是否仍然保持其初始特征，特别是致病性，是至关重要的。有一种可能性，不适当的氯化可能会刺激存活细菌的致病性，由于诱导转录的基因编码的毒力，活性氧（ROS）的反应，或毒力基因的水平转移。本项目旨在通过研究宿主-细菌相互作用，更好地了解氯化后存活微生物的致病性。具体而言，本项目旨在：1）考察氯应激细菌在模拟胃肠道条件下的耐受性; 2）检测氯应激细菌对人肠上皮细胞的粘附和侵袭; 3）比较细胞在暴露于非应激和氯应激细菌下的炎症反应; 4）使用多重qPCR阵列确定毒力基因表达的变化; 5）通过全基因组转录分析阐明氯胁迫细菌对氯的细胞反应。该研究金项目的预期成果将有助于开发新的消毒策略，以降低存活细菌的致病性，并最大限度地减少具有致病性的细菌在配水系统中的传播。该方法可推广到食品防腐和医用消毒剂等领域。EPSCoR RII Track-4：NSF计划的支持将使PI的机构能够提高其研究能力并与EPA建立合作网络。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Bacterial pathogens in water pose a significant worldwide threat to public health, especially for individuals with weakened immune systems. Currently, chlorination is the most widely used water disinfection process to cause the inactivation of microorganisms in the water to ensure public and environmental health. While most bacteria in water can be effectively killed after chlorination, some may survive due to their resistance to disinfectants. There has yet to be a comprehensive study to investigate the ability of the surviving bacteria to infect a host and cause disease. This Research Infrastructure Improvement RII Track-4 fellowship project will explore the impact of water disinfection on the pathogenicity of bacteria, which will help overcome the current knowledge gap regarding microbial risk assessment after water treatment. This goal will be addressed by using advanced cellular tools and next-generation sequencing techniques developed at the host site, the Biological Measurements Branch of the US Environmental Protection Agency. The fundamental knowledge obtained from this project is expected to provide valuable information to stakeholders and policymakers for reassessing the current water treatment processes and reducing health risks. This research will also offer training opportunities for underrepresented students at New Mexico State University, which is an Hispanic-Serving Institution.This Research Infrastructure Improvement Track-4 EPSCoR Research Fellows (RII Track-4) proposal would provide a fellowship to an Assistant Professor and training for a graduate student at New Mexico State University. This work would be conducted in collaboration with researchers in the Biological Measurements Branch of the Watershed & Ecosystem Characterization Division at the United States Environmental Protection Agency (EPA) in Cincinnati, OH. Knowing if the surviving bacteria after disinfection still hold their initial characteristics, especially pathogenicity, is essential. There is a possibility that inappropriate chlorination may stimulate the pathogenicity of surviving bacteria due to the induced transcription of gene encoding virulence, reactive oxygen species (ROS) response, or horizontal transfer of virulence genes. This project aims to better understand the pathogenicity of surviving microorganisms after chlorination by investigating host-bacteria interactions. Specifically, this project aims to: 1) Investigate the tolerance of chlorine-stressed bacteria under simulated gastrointestinal conditions; 2) Examine the adhesion and invasion of chlorine-stressed bacteria to human intestinal epithelial cells; 3) Compare the inflammatory responses of cells under exposure to non-stressed and chlorine-stressed bacteria; 4) Determine the changes in virulence gene expression using multiple qPCR arrays; and 5) Elucidate cellular response of chlorine-stressed bacteria to chlorine through genome-wide transcriptional analysis. The expected outcomes of this fellowship project will facilitate the development of novel disinfection strategies to reduce the pathogenicity of surviving bacteria and minimize the spread of bacteria with pathogenicity into the water distribution system. The methods developed in the proposed study could be expanded to the fields of food preservation and medical disinfectants. The support from the EPSCoR RII Track-4:NSF program will allow the PI's institution to advance its research capacity and establish collaboration networks with EPA. It also provides a foundation for building the PI's research program into a significant and regular contributor to pathogen removal in water systems.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.