Pilot: Center for Appalachian Research in Environmental Sciences: Wastewater Surveillance of Water and Vector-Borne Pathogens Following Climate Disasters in Eastern Kentucky

ABSTRACT Climate change is expected to increase the temperature of surface water reservoirs and weather patterns in Kentucky. Many regions of eastern Kentucky (EKY) are susceptible to severe flooding which may require weeks of emergency service operations to restore basic needs for families. During the initial storm surge, overwhelmed sewage infrastructure contaminates the local environment while exposing emergency personnel and fleeing survivors to various pathogens. Gastrointestinal pathogens, such as Vibrio cholerae, Vibrio vulnificus, and Salmonella enterica, replicate faster in warm brackish waters following extreme temperature and rainfall events. Endemic rodent and mosquito-borne diseases are known to increase in the following weeks as flood waters recede. Microbial and chemical contamination following heavy rain is not limited to coastal regions, as shown by a 2011 study along the Ohio River that found higher levels of various pathogens in surface water after a flood. However, given that many microbes (e.g., Vibrio, Escherichia) exist in both pathogenic and non-pathogenic states, additional assay specificity is required to identify toxin genes related to disease6 and to better understand the lasting environmental impact of more frequent natural disasters. We aim to identify these pathogens during a flood to better inform public health responses to inevitable climate-disasters. Further, given the dynamic and unpredictable nature of post-disaster settings, assays must be easy-to-deploy, robust, and rapid. To overcome these challenges, we plan to adapt a suite of technologies/workflows that we developed for simple and rapid wastewater-based epidemiology (WBE), with the goal of enabling simple/rapid analyses of pathogens and pathogen biomarkers in flood waters. Our WBE work involves a van-based laboratory that circulates throughout Appalachia to provide rapid, point-of-sample measurements, with the aid of citizen scientists to accurately collect samples and perform assays. Our mobile van can collect, concentrate, and analyze wastewater using either PCR and/or long-read Oxford Nanopore Technologies (ONT) sequencing. Metagenomic ONT sequencing will be our primary data analysis endpoint due to the breadth of metagenomic analyses (i.e., pathogens need not be known a priori as with PCR) and the simple/low-cost nature of our ONT-based workflow. Our preliminary ONT sequencing data suggests that measurable pathogen biomarkers change over time and are highly region specific. We hypothesize that low-cost sample collection and sequencing are capable of identifying pathogen load in water reservoirs immediately following a flood and these levels can be temporally tracked over the course of the disaster.