Phase I SBIR proposals will be accepted. Fast-Track proposals will not be accepted. Phase I clinical trials will not be accepted. Number of anticipated awards: 1-2 Budget (total costs): Phase I: up to $243,500 for up to 6 months; Phase II of up to $1,000,000 and a Phase II duration of up to 2 years PROPOSALS THAT EXCEED THE BUDGET OR PROJECT DURATION LISTED ABOVE MAY NOT BE FUNDED. Background Wastewater surveillance provides a powerful independent approach to complement existing surveillance systems. Wastewater surveillance is currently being used to support the COVID-19 response. SARS-CoV-2 RNA is shed in the feces of individuals with both symptomatic and asymptomatic infections and SARS-CoV-2 RNA in wastewater has been demonstrated to be a leading indicator of reported cases and hospitalizations. There is no evidence to date that anyone has become sick with COVID-19 because of exposure to wastewater, but wastewater is a hazardous material and molecular analyses of samples that could potentially contain multiple human pathogens are severely restricted. This results in significant barriers for laboratories onboarding testing and there is a need for a product or procedure to provide molecular preservation (stabilization) and pathogen inactivation from post-sampling of wastewater to help overcome these barriers. Project Goals The goal of the proposed research is to develop a product to inactivate and stabilize wastewater samples for shipping and transport. This product must be amenable to on-site use by stakeholders, such as State Agencies for wastewater monitoring, etc., and could be physical or chemical inactivation. The product can be an all-encompassing portable sampler that inactivates and provides molecular preservation of pathogens in wastewater or be used sequentially with existing samplers without adding additional biosafety risks to the collection procedure. The product will provide a qualitative indicator that the inactivation process has occurred, and the inactivation of the wastewater sample must not interfere with downstream molecular testing. Phase I Activities and Expected Deliverables The expected deliverables are: 1. Develop or adapt a method to inactivate and molecularly preserve SARS-CoV-2, or the proxy virus controls, bovine coronavirus (BCoV), murine coronavirus (MCoV, e.g., murine hepatitis virus), bacteriophage Phi6, or human coronavirus OC43, in wastewater samples; the method must be able to provide an indicator that the inactivation process has occurred. 2. Quantify molecular detection before and after the molecular preservation and inactivation procedure for SARSCoV-2, or the proxy virus controls bovine coronavirus (BCoV), murine coronavirus (MCoV, e.g., murine hepatitis virus), bacteriophage Phi6, or human coronavirus OC43, in wastewater samples. Method development with surrogate viruses will be considered but final evaluation with SARS-CoV-2 must be included. Page 127 3. Conduct matrix evaluation to understand the assay performance using different wastewater types (e.g., raw wastewater, sludge). Impact The product of this proposed research will allow laboratories to test and monitor their wastewater supplies for SARS-CoV-2 and other emerging pathogens without the need for dedicated containment laboratories, which is currently not possible. Once safe processing is available, laboratory capacity can scale up and expand to other pathogens that pose a public health threat and thereby inform control processes and ultimately reduce the burden of infections. Commercialization Potential This research will lead to the development of new products that inactivate infectious material in wastewater samples and provide molecular preservation to benefit stakeholders at every point in the wastewater surveillance process from collection to testing. Potential products include inactivation and stabilization systems, inactivation indicator kits, storage and transport products, and wastewater sampling devices that provide an all-in-one “sample collection-to-inactivated and stabilized infectious material” transport sample container. These products could be used by water managers, universities, businesses, correctional facilities, and healthcare facilities, as well as federal, state, and local public and environmental health agencies. The market for products for wastewater surveillance sample collection and testing has grown exponentially during the COVID-19 pandemic and is likely to continue to grow as public health departments establish longer-term wastewater-based disease surveillance programs for SARSCoV-2 and other disease targets.