Using ELISA to detect pathogen antibodies in wild mammal carcasses: a systematic literature review

Carcasses of wild mammals provide convenient samples for wildlife disease investigators. Specifically, antibodies can persist in the blood and tissues of animals up to a few days after death and can provide information on exposure to specific pathogens, including microorganisms with zoonotic implications, such as Toxoplasma gondii, Trichinella spp., Brucella spp., and Mycobacterium spp. Although screening samples from decaying wildlife for antibodies is not a new method, an overview of the literature with clear recommendations on best practices is lacking. Here, we present results from a systematic review of 115 studies using enzyme-linked immunosorbent assay (ELISA) to detect pathogen antibodies in wild mammals across 298 individual research projects. We discuss the range of mammalian hosts and the pathogens most represented in the literature and suggest improvements to consider when using samples from decaying animals for seroinvestigation. Our results indicate that the most examined mammals were artiodactyl ungulates and that antibodies against the bacteria Brucella spp. are those most examined to date, followed by the protozoan Toxoplasma gondii, malignant catarrhal fever virus, Schmallenberg orthobunyavirus, and the coccidian Neospora caninum. The effect of haemolysis in serum samples on ELISA results was considered in only 3.7% of the research projects we reviewed, sample size decisions were not outlined in 83% of research projects, and half of the projects did not examine seroprevalence in different age classes. Nevertheless, these details are important for replicability and accurate interpretation of results. We highlight the benefits and limitations of detecting antibodies against pathogens in carcasses from wild mammalian groups. Wild mammals are reservoirs for a wide range of pathogens including those with zoonotic potential and investigating their carcasses can help wildlife investigators to better understand the epidemiology of many diseases with implications for wildlife, livestock, and human health.