Experimental Susceptibility of North American Raccoons (Procyon lotor) and Striped Skunks (Mephitis mephitis) to SARS-CoV-2

Recent spillback events of SARS-CoV-2 from humans to animals has raised concerns about it becoming endemic in wildlife. A sylvatic cycle of SARS-CoV-2 could present multiple opportunities for repeated spillback into human populations and other susceptible wildlife. Based on their taxonomy and natural history, two native North American wildlife species —the striped skunk (Mephitis mephitis) and the raccoon (Procyon lotor) —represent a high likelihood of susceptibility and ecological opportunity of becoming infected with SARS-CoV-2. Eight skunks and raccoons were each intranasally inoculated with one of two doses of the virus (103 PFU and 105 PFU) and housed in pairs. To evaluate direct transmission, a naïve animal was added to each inoculated pair 48 h post-inoculation. Four control animals of each species were handled like the experimental groups. At predetermined intervals, we collected nasal and rectal swabs to quantify virus shed via virus isolation and detect viral RNA via rRT-PCR and blood for serum neutralization. Lastly, animals were euthanized at staggered intervals to describe disease progression through histopathology and immunohistochemistry. No animals developed clinical disease. All intranasally inoculated animals seroconverted, suggesting both species are susceptible to SARS-CoV-2 infection. The highest titers in skunks and raccoons were 1:128 and 1:64, respectively. Low quantities of virus were isolated from 2/8 inoculated skunks for up to day 5 post-inoculation, however no virus was isolated from inoculated raccoons or direct contacts of either species. Neither species had gross lesions, but recovering mild chronic pneumonia consistent with viral insult was recorded histologically in 5/8 inoculated skunks. Unlike another SARS-CoV-2 infection trial in these species, we detected neutralizing antibodies in inoculated raccoons; thus, future wildlife serologic surveillance results must be interpreted with caution. Due to the inability to isolate virus from raccoons, the lack of evidence of direct transmission between both species, and low amount of virus shed by skunks, it seems unlikely for SARS-CoV-2 to become established in raccoon and skunk populations and for virus to spillback into humans. Continued outbreaks in non-domestic species, wild and captive, highlight that additional research on the susceptibility of SARS-CoV-2 in wildlife, especially musteloidea, and of conservation concern, is needed.

Resistance Patterns, mcr-4 and OXA-48 Genes, and Virulence Factors of Escherichia coli from Apennine Chamois Living in Sympatry with Domestic Species, Italy

The aim of this study was to determine and characterize potential resistance mechanisms against selected Critically Important Antibiotics in Escherichia coli isolates collected from wild and domestic ruminants living in the Maiella National Park, in Central Italy. A total of 38 isolates were obtained from red deer, Apennine chamois, cattle, sheep, and goats grazing in lands with different levels of anthropic pressure. Antimicrobial susceptibility was determined by Minimal Inhibitory Concentration testing, showing phenotypic resistance to colistin, meropenem, or ceftazidime in 9 isolates along with one bacterial strain being resistant to three of the tested antibiotics. In addition, the biomolecular assays allowed the amplification of the genes conferring the colistin (mcr-4), the carbapenems (OXA-48), penicillins and cephalosporins (TEM, SHV, CMY-1, CMY-2) resistance. In order to describe the potential pathogenicity of isolates under study, virulence genes related to Shiga toxin-producing (STEC) and enteropathogenic (EPEC) pathovars were identified. This study is the first report of mcr-4 and OXA-48 genes in resistant E. coli harboring virulence genes in Italian wildlife, with special regard to Apennine chamois and red deer species. The multidisciplinary approach used in this study can improve the early detection of emerging antibiotic resistance determinants in human-animal-environment interfaces by means of wildlife monitoring.

A Novel Terrestrial Rabies Virus Lineage Occurring in South America: Origin, Diversification, and Evidence of Contact between Wild and Domestic Cycles

The rabies virus (RABV) is characterized by a history dominated by host shifts within and among bats and carnivores. One of the main outcomes of long-term RABV maintenance in dogs was the establishment of variants in a wide variety of mesocarnivores. In this study, we present the most comprehensive phylogenetic and phylogeographic analysis, contributing to a better understanding of the origins, diversification, and the role of different host species in the evolution and diffusion of a dog-related variant endemic of South America. A total of 237 complete Nucleoprotein gene sequences were studied, corresponding to wild and domestic species, performing selection analyses, ancestral states reconstructions, and recombination analyses. This variant originated in Brazil and disseminated through Argentina and Paraguay, where a previously unknown lineage was found. A single host shift was identified in the phylogeny, from dog to the crab-eating fox (Cerdocyon thous) in the Northeast of Brazil. Although this process occurred in a background of purifying selection, there is evidence of adaptive evolution -or selection of sub-consensus sequences- in internal branches after the host shift. The interaction of domestic and wild cycles persisted after host switching, as revealed by spillover and putative recombination events.

Efficacy and Function of Feathers, Hair, and Glabrous Skin in the Thermoregulation Strategies of Domestic Animals

The objective of this review is to describe and analyze the effect of feathers, hair, and glabrous (hairless) skin on the thermoregulation of domestic and endotherm animals, especially concerning the uses and scope of infrared thermography (IRT), scientific findings on heat and cold stress, and differences among species of domestic animals. Clinical medicine considers thermoregulation a mechanism that allows animals to adapt to varying thermal environmental conditions, a process in which the presence of feathers, hair, or glabrous skin influences heat loss or heat retention, respectively, under hot and cold environmental conditions. Evaluating body temperature provides vital information on an individual’s physiological state and health status since variations in euthermia maintenance in vertebrates reflect a significant cellular metabolism deviation that needs to be assessed and quantified. IRT is a non-invasive tool for evaluating thermal responses under thermal stress conditions in animals, where the presence or absence of feathers, hair, and glabrous skin can affect readings and the differences detected. Therefore, anatomical regions, the characteristics of feathers, hair, glabrous skin such as structure, length, color, and extension, and strategies for dissipating or retaining heat together constitute a broad area of opportunity for future research into the phenomena of dermal thermoregulation in domestic species.

Of Mice, Cattle, and Men: A Review of the Eco-Epidemiology of Leptospira borgpetersenii Serovar Ballum

In New Zealand (NZ), leptospirosis is a mostly occupational zoonosis, with >66% of the recently notified cases being farm or abattoir workers. Livestock species independently maintain Leptospira borgpetersenii serovar Hardjo and L. interrogans serovar Pomona, and both are included in livestock vaccines. The increasing importance in human cases of Ballum, a serovar associated with wildlife, suggests that wildlife may be an overlooked source of infection. Livestock could also act as bridge hosts for humans. Drawing from disease ecology frameworks, we chose five barriers to include in this review based on the hypothesis that cattle act as bridge hosts for Ballum. Using a narrative methodology, we collated published studies pertaining to (a) the distribution and abundance of potential wild maintenance hosts of Ballum, (b) the infection dynamics (prevalence and pathogenesis) in those same hosts, (c) Ballum shedding and survival in the environment, (d) the exposure and competency of cattle as a potential bridge host, and (e) exposure for humans as a target host of Ballum. Mice (Mus musculus), rats (Rattus rattus, R. norvegicus) and hedgehogs (Erinaceus europaeus) were suspected as maintenance hosts of Ballum in NZ in studies conducted in the 1970s–1980s. These introduced species are distributed throughout NZ, and are present on pastures. The role of other wildlife in Ballum (and more broadly Leptospira) transmission remains poorly defined, and has not been thoroughly investigated in NZ. The experimental and natural Ballum infection of cattle suggest a low pathogenicity and the possibility of shedding. The seroprevalence in cattle appears higher in recent serosurveys (3 to 14%) compared with studies from the 1970s (0 to 3%). This review identifies gaps in the knowledge of Ballum, and highlights cattle as a potential spillover host. Further studies are required to ascertain the role that wild and domestic species may play in the eco-epidemiology of Ballum in order to understand its survival in the environment, and to inform control strategies.

Medium-sized and large mammals of the Floresta da Cicuta Area of Relevant Ecological Interest, a protected area in southeastern Brazil

The Atlantic Forest is one of the most biodiverse biomes in the world and has been severely degraded and fragmented, with the extirpation of most medium-sized and large vertebrates from the forest remnants. Here we present the results of a survey of medium-sized and large mammals in an area of protected seasonal semideciduous forest, the Floresta da Cicuta Area of Relevant Ecological Interest (ARIE-FC), Rio de Janeiro, Brazil, part the Atlantic Forest biome. We used camera traps (2,257 camera days) and direct observations over a 23-month period. We recorded 19 species (including two domestic species), seven of which are classified as at-risk, such as Leopardus guttulus (Hensel, 1872), Sylvilagus tapetillus Thomas, 1913, Alouatta clamitans Cabrera, 1940, and Chrysocyon brachyurus (Illiger, 1815). A diverse terrestrial mammal assemblage in the ARIE-FC reinforces the importance of small forest fragments for the conservation of biodiversity in human-modified landscapes of the Brazilian Atlantic Forest.

258 W112 Reproductive Performance in Domestic Ruminants: 50 Years of accomplishment: a USDA-NIFA Multistate Research Project

This symposium acknowledges the founders of W112 Reproductive Performance in Domestic Ruminants. The contributions they made laid foundation collaborative efforts that continued over the next half century. The W112 really began in 1969 as a rewritten project involving several Agricultural Experiment Stations. That original project was W-49, that had been in existence at least as early as 1958. Upon approval of the project renewal in early 1970, the project was renamed W112 Reproductive Performance in Domestic Ruminants. The list of names of participants is like a who’s who of reproductive physiologists, too long to enumerate here. Increasing knowledge of physiology and endocrinology were paramount during the early years. Development, improvement, and sharing of analytical techniques improved accuracy of results, safety of procedures, and knowledge to be gained from the research. Concurrently, mechanistic basic and applied studies involving oocyte development, ovulation, sperm development, fertilization, pregnancy, and parturition all brought new understanding of the reproductive biology of domestic species. A good deal of what we currently know of the reproductive biology of ruminant animals finds some basis in laboratories involved in W112 collaborations. The unique nature of these scientists resulted from their mutual respect. Many young scientists found mentors in W112 that assisted us in developing our own research programs. And the support has not ended with research collaboration. It has been common for project participants to share teaching resources and serve on graduate student committees for each other’s students, fostering future involvement in the project. The W112 began slightly more than 50 years ago. Those of us who have been part of this group share a hope that the multistate project named Reproductive Performance in Domestic Ruminants will still be meeting the same goals of collaboration and support among scientists working to further animal agriculture for another 50 years and more.

Cranial shape diversification in horses: variation and covariation patterns under the impact of artificial selection

The potential of artificial selection to dramatically impact phenotypic diversity is well known. Large-scale morphological changes in domestic species, emerging over short timescales, offer an accelerated perspective on evolutionary processes. The domestic horse (Equus caballus) provides a striking example of rapid evolution, with major changes in morphology and size likely stemming from artificial selection. However, the microevolutionary mechanisms allowing to generate this variation in a short time interval remain little known. Here, we use 3D geometric morphometrics to quantify skull morphological diversity in the horse, and investigate modularity and integration patterns to understand how morphological associations contribute to cranial evolvability in this taxon. We find that changes in the magnitude of cranial integration contribute to the diversification of the skull morphology in horse breeds. Our results demonstrate that a conserved pattern of modularity does not constrain large-scale morphological variations in horses and that artificial selection has impacted mechanisms underlying phenotypic diversity to facilitate rapid shape changes. More broadly, this study demonstrates that studying microevolutionary processes in domestic species produces important insights into extant phenotypic diversity.

Hepatitis E Virus seroprevalence among cows in a rural area of southern Italy

Hepatitis E virus (HEV) is considered to be an emerging zoonotic disease, which causes numerous cases of hepatitis and deaths worldwide each year. Pigs are a host reservoir of HEV, but numerous other wild and domestic species can be infected. The aim of this study was to carry out serological screening of anti-HEV antibodies in cattle in the Sicily region (Southern Italy). Between April and December 2018, 231 serum samples were collected from cows and analysed with enzyme-linked immunosorbent assay (ELISA), for detection of HEV antibodies (IgG). The overall prevalence of anti-HEV antibodies was 36.36% (84/231; 95% CI 30-43). There were statistically significant differences in the HEV seroprevalence in animals of different ages (<11 months: 11.41%; >12 months: 39.30%; >24 months: 54.10%) (P<0.05). No statistically significant differences were detected between genders (31.37% in male and 37.38% in female) (P >0.05). This study shows that further investigation of HEV in cattle is required to understand better the epidemiology in farm animals and the potential zoonotic risks for humans. This is the first report of HEV seroprevalence for cows in Italy, which provides baseline data for further studies and for control of HEV infection in cattle.