Microbe surveillance in the amphibian pet trade: Results from a pilot study

Regional and global trade of live animals can contribute to the spread and emergence of novel pathogens, including several important pathogens of amphibians. However, understanding the spread or even frequency of infections in large, complex amphibian trade networks has been difficult, in part because businesses tend to be reluctant to participate in surveillance programs. Thus, we developed a novel approach to surveillance in which anonymous participating businesses were sent surveillance kits through a trusted trade advocacy partner, samples were returned to researchers via anonymous prepaid envelopes, and results were provided via a secure website with access regulated by a unique personal identification number (PIN) created by the business. We tested samples for the amphibian pathogens, Batrachochytrium salamandrivorans (Bsal), Batrachochytrium dendrobatidis (Bd), and Ranavirus spp. (Rv), as well as the beneficial microbe, Janthinobacterium lividum (Jliv), using quantitative real-time polymerase chain reaction (qPCR). Out of 120 businesses invited to complete an anonymous socioeconomic survey, 24 volunteered to participate in pathogen surveillance, of which 14 were sent surveillance kits. Eight of these businesses returned samples consisting of swabs collected from amphibians in 78 terrestrial habitats and water filters from 49 aquatic habitats. Copies of a highly conserved vertebrate gene (EBF3N), quantified using qPCR, were consistently low (<100 copies) in returned samples, but similar to those collected by researchers, indicating comparable sample quality. Three samples (from two facilities) had detectable levels of Bd DNA; Bsal, Rv, and Jliv were not detected. This pilot study provides evidence that information about pathogens in pet trade networks can be acquired by developing partnerships with industry, and business participation might be enhanced by ensuring anonymity and inclusion of a trade advocacy partner.