Beyond the Host: Evaluating Methods for Environmental Biosurveillance Ofviruses Using Avian Influenza Virus as a Case Study

Environmental DNA and RNA-based virus biosurveillance holds promise as an early warningsystem for detecting pathogen incursions and tracking their evolution across fields. Unliketraditional pathogen monitoring, which typically relies on direct sampling from humans and animalhosts, environmental biosurveillance offers a broader, faster, more accessible, and cost-effectiveapproach. While recent advancements in viral detection methods have expanded their utility, therelative effectiveness and suitability of these methods for point-of-need detection remain unclear.To this end, we evaluate and compare 12 contemporary viral detection methods to assess theirpotential for identifying viruses in environmental samples, with a specific focus on highlypathogenic avian influenza (HPAI) virus. We examine the scalability, practicality, and futurepotential of each method, particularly those that have yet to be applied to environmental samples,while considering the specific requirements for each approach. Our goal is to provide guidance toend-users on selecting the most appropriate methods based on current available approaches anddata. In addition, we outline key requirements for an “optimal” in-field detection method andidentify the steps necessary to enhance environmental virus biosurveillance in the future.