key: cord-0749848-thmbciel authors: Bosco-Lauth, Angela M.; Root, J. Jeffrey; Porter, Stephanie M.; Walker, Audrey E.; Guilbert, Lauren; Hawvermale, Daphne; Pepper, Aimee; Maison, Rachel M.; Hartwig, Airn E.; Gordy, Paul; Bielefeldt-Ohmann, Helle; Bowen, Richard A. title: Survey of peridomestic mammal susceptibility to SARS-CoV-2 infection date: 2021-01-21 journal: bioRxiv DOI: 10.1101/2021.01.21.427629 sha: 45f5d0ea31e658900e4cba040aaf9b2f29b738aa doc_id: 749848 cord_uid: thmbciel Wild animals have been implicated as the origin of SARS-CoV-2, but it is largely unknown how the virus affects most wildlife species and if wildlife could ultimately serve as a reservoir for maintaining the virus outside the human population. Here we show that several common peridomestic species, including deer mice, bushy-tailed woodrats, and striped skunks, are susceptible to infection and can shed the virus in respiratory secretions. In contrast, we demonstrate that cottontail rabbits, fox squirrels, Wyoming ground squirrels, black-tailed prairie dogs, house mice, and racoons are not susceptible to SARS-CoV-2 infection. Our work expands upon the existing knowledge base of susceptible species and provides evidence that human-wildlife interactions could result in continued transmission of SARS-CoV-2. The rapid global expansion of severe acute respiratory syndrome (SARS) coronavirus 2 35 (SARS-CoV-2) has been unprecedented in modern history. While the original human 36 infection(s) were potentially linked to wild animals in a wet market (1) , human-to-human 37 transmission is currently the dominant mechanism of viral spread. Peridomestic animals, which 38 are represented by wild and feral animals living within close proximity to humans, represent key 39 species to evaluate for SARS-CoV-2 epidemiology for multiple reasons. First, given their 40 common associations with humans and anthropogenically modified habitats, they represent the 41 wildlife species with the greatest chance of exposure to the virus from humans (i.e., reverse 42 zoonosis) or pets such as cats. Second, should select peridomestic wildlife prove to be 43 susceptible to the virus and have the capacity to replicate it to high viral titers, these species 44 would have the potential to maintain the virus among conspecifics. Third, should some species 45 possess the maintenance host criteria mentioned above, they would represent wildlife species 46 that would have the greatest chance (e.g., shedding ability and proximity to humans) to spread 47 the virus back to humans. Wild rodents, cottontail rabbits (Sylvilagus sp.), raccoons (Procyon 48 lotor) and striped skunks (Mephitis mephitis) can exhibit peridomestic tendencies in urban and 49 suburban environments. Members of all these species/taxonomic groups have been shown to 50 shed influenza A viruses following experimental inoculations (2,3,4), suggesting they might 51 harbor productive infections when exposed to other human-pathogenic respiratory viruses. Based upon protein analyses of amino acid residues of ACE2, TMPRSS2 and S protein, 54 species susceptibility analyses suggested that, among other taxonomic groups, both carnivores 55 and wild rodents are potentially high-risk groups (5,6,7). Predicting specific species' 56 susceptibility, however, is more challenging. Looking at protein sequence analysis of ACE2 57 binding with the S protein of SARS-CoV-2, one study indicated that raccoons could be ruled out 58 as potential hosts for SARS-CoV-2 (6) and a different study based upon sequence analysis 59 suggested that the western spotted skunks (Spilogale gracilis) had a very low prediction of 60 SARS-CoV-2 S-binding propensity (7). Similarly, the same study also suggested that American 61 mink (Neovison vison) have a similar prediction as western spotted skunks (7). However, over 62 the last several months, outbreaks of SAR-CoV-2 in commercial mink farms have been noted in 63 Europe and more recently in the U.S. (8, 9) . Respiratory problems, rapid transmission, and/or 64 unusually high mortality have been noted in this species in various regions (9,10), which 65 suggests that the aforementioned analyses have limitations. Rodents are the largest and most diverse order of mammals, so it is unsurprising that the 68 susceptibility of rodents to SARS-CoV-2 varies by species. there are more than 1700 species of rodents world-wide, many of which exist closely at the Further, as with rodents, wild rabbits are highly likely to be predated upon by domestic cats. In this study, we assessed six common peridomestic rodent species for susceptibility to 104 SARS-CoV-2: deer mice, wild-caught house mice (Mus musculus), bushy-tailed woodrats (aka 105 "pack rats"; Neotoma cinerea), fox squirrels (Sciurus niger), Wyoming ground squirrels The following mixed-sex animals were evaluated for susceptibility to SARS-CoV-2: Deer mice, 119 house mice, bushy-tailed woodrats, Wyoming ground squirrels, black-tailed prairie dogs, fox 120 squirrels, cottontail rabbits, striped skunks, and raccoons. Deer mice, house mice and bushy-121 tailed woodrats were trapped using Sherman traps baited with grain. Wyoming ground squirrels, 122 fox squirrels, black-tailed prairie dogs, and cottontails were trapped using Tomahawk live traps 123 (e.g., 7 x 7 x 20 or 7 x 7 x 24). All trapping was done in Northern Colorado (Larimer, Jackson Groups of three animals from each species (two for ground squirrels) were used for preliminary 157 studies to evaluate viral shedding and acute pathological changes. For these animals, oral swabs were obtained pre-challenge and on days 1-3 post-challenge, at which time animals were 159 euthanized and the following tissues harvested for virus isolation and formalin fixation: trachea, 160 nasal turbinates, lung, heart, liver, spleen, kidney, small intestine, and olfactory bulb. The 161 exception to this was raccoons, for which only one animal was euthanized at day 3; the 162 remaining two were kept through day 28 to evaluate serological response. The remaining 3-6 163 animals per select species were swabbed daily from days 0-5 and 7 to further evaluate duration Table 1 illustrates the necropsy scheme for each species. 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