Surveillance of wildlife and livestock for novel and emerging coronaviruses

Project Summary There is an urgent need for coronavirus (CoV) surveillance in both wildlife and livestock. As seen repeatedly, CoVs have a propensity to jump species from their bat, rodent and avian reservoirs, often to intermediate species including livestock, before infecting humans. Our knowledge of North American CoVs in wildlife species is sparse and routine sequencing of livestock CoVs are limited. In our first aim, to detect and sequence coronaviruses in wildlife and livestock, we will utilize a pan-coronavirus RT-PCR to screen a unique sample set consisting of bats and wildlife rarely surveilled. The sample set is comprised of ~500 animals submitted to our veterinary diagnostic laboratory for rabies testing over a 12-month period and consists of pooled lung, spleen and intestine tissue homogenate. In parallel, we will screen food animal diagnostic submissions from animals with acute enteric and respiratory disease. Our second aim, to further characterize coronavirus genomes and characterize virus isolates, phylogenetic and recombination analysis will be performed with the assembled CoV genomes. Specifically, assembled genomes will be annotated and submitted to Genbank and made publicly available immediately. Phylogenetic analysis will determine if SARS-CoV-2, CoVs similar to SARS-CoV-2 or novel CoVs have emerged in non-human species. Recombination analysis will next be performed using SARS-CoV-2 and the assembled CoV genomes to determine if SARS-CoV-2 or related CoVs have undergone natural recombination and contributed genetic material to endemic CoV. Another component of this aim is to isolate and propagate the detected CoVs in cell culture. This latter aim is critical for further pathogenesis studies in the natural host and model organisms. This R21 exploratory project meets an urgent need for CoV surveillance at the human-animal interface, both in wildlife and livestock. Results from this study will identify emerging disease threats and develop critical tools, including sequences, detection methodology and cell culture adapted viruses that will enable further studies on possible zoonosis and enable preemptive action.