The goal of this project was to develop simple and rapid methods for virus detection in water to aid US marine mammal program veterinarians in caring for bottlenose dolphins (Tursiops truncates) and California sea lions (Zalophus californianus). These marine mammals need to be protected from viral diseases while they stay in a netted enclosure in San Diego Bay, California. The purpose of this study was to evaluate the Innovaprep Concentrating Pipette Select (CP Select) for the rapid concentration of viruses in environmental water samples. Bacteriophages MS2, P22, Phi6, and PhiX-174 were seeded in artificial seawater and tested for recovery after using the CP Select. These bacteriophages vary in characteristics and were selected to be surrogates for animal viruses (e.g., Levivirdae, Podoviridae, Cystoviridae, and Microviridae). Recovery for all bacteriophages was evaluated either using a plaque assay and/or Droplet Digital PCR (ddPCR). After a method was established using phages, three animal viruses: adenovirus (Adenoviridae), coronavirus OC43 (Coronaviridae), and morbillivirus canine distemper virus (CDV, Paramyxovirus) were included in this study using artificial seawater as well as water from the John Shedd Aquarium and San Diego Bay. These animal viruses were quantified using ddPCR. The culture assay average recovery for each phage was: 82.73 ±27.3% for P22 (n=6), 4.84 ±3.8% for MS2 (n=3), 32.77 ±19.3% for PhiX174 (n=6), and 71.48 ±42.5% for Phi6 (n=6). The ddPCR assay average recovery was 7.82 ±8.1% for MS2 (n=3), 34.72 ±19.7% for Phi6 (n=6), 39.31 ±26.6% for Adenovirus (n=9), 19.04 ±11.6% for OC43 (n=9), and 19.84 ±13.6% for CDV (n=9). In the future, this method will be shared with the US marine mammal program so that they may implement surveillance in their netted enclosure. This method could also be implemented in aquaria or at public beaches for water quality testing.
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