Didelphis Virginiana Research Articles

We are not the problem, cars are: effect of different human activities in an urban area on opossums health and population size

AbstractUnderstanding the health of urban animals is key for preventing and controlling zoonotic diseases. To achieve this goal, it’s necessary to understand what factors could have an impact on those animals’ health. Therefore, the aim of this article is to compare some health characteristics of opossums (Didelphis virginiana) in urban environments with and without human presence. For this, we trapped opossums in two urbanized areas: one dominated by buildings and one by roads and parking lots (plus two control locations in natural areas). During two periods: normal activities and under lockdown due to COVID-19 conditions (when the presence of people was minimal). We found that in the building area, human presence has almost no effect on the opossums’ health (with only minor changes in erythrocyte counts and tar coverage); however, in the parking lot, the opossum population dropped drastically because of human activities (use of vehicles). This suggests that human presence is not harmful for those animals, but cars are. Therefore, the use of vehicles has a negative impact on opossums’ health and could increase the spreading of zoonotic diseases.

Human-derived food shrinks home ranges and alters resource selection of mammals at the urban-wild interface

Protected lands are an important source of food, shelter, and reproductive opportunities for wildlife, especially in urban landscapes. When urban development abuts the edges of undisturbed ecosystems, synanthropic species can alter their foraging behaviors and movement to utilize human-supplemented resources throughout the urban-wild interface. Therefore, urban development on the edges of protected lands can have pronounced effects on animal movement and ecosystem functions. Iconic urban adaptive mesopredators such as northern raccoons (Procyon lotor) and Virginia opossums (Didelphis virginiana) often benefit from human-supplemented food sources such as unsecured garbage, pet food, and fresh water when available. To investigate how urban edges affect the movements of urban-adapted omnivores within conservation lands, we estimated home ranges and third-order resource selection of 27 raccoons and 12 opossums with GPS collars throughout the protected areas of northern Key Largo, FL, USA between April 2022–October 2023. The proportion of urban development in an individual's home range was the most influential factor associated with home range size, followed by species and sex. Individuals with greater proportions of residential neighborhoods and commercial areas in their home ranges exhibited smaller home ranges. Third-order resource selection functions identified both mesopredator species using residential and commercial land use areas more than they were available on the landscape. These results indicate that urban areas attract urban-adapted mesopredators from protected areas and result in smaller home ranges in the face of abundant human-derived food. Reduced home ranges on edges can support higher densities of animals, which may increase rates of disease transmission, especially when the urban borders support populations of feral domestic species. Shifting foraging behaviors from the protected areas to urban edges could have cascading downward effects if seed-dispersing roles are diluted. As urbanization increases and the distance between wild lands and human disturbance decreases, it is increasingly important to study the mechanisms of how urban development on the edges of protected areas affect the movement of wildlife.

A new biomechanical model of the mammal jaw based on load path analysis.

The primary function of the tetrapod jaw is to transmit jaw muscle forces to bite points. The routes of force transfer in the jaw have never been studied but can be quantified using load paths - the shortest, stiffest routes from regions of force application to support constraints. Here, we use load path analysis to map force transfer from muscle attachments to bite point and jaw joint, and to evaluate how different configurations of trabecular and cortical bone affect load paths. We created three models of the mandible of the Virginia opossum, Didelphis virginiana, each with a cortical bone shell, but with different material properties for the internal spaces: (1) a cortical-trabecular model, in which the interior space is modeled with bulk properties of trabecular bone; (2) a cortical-hollow model, in which trabeculae and mandibular canal are modeled as hollow; and (3) a solid-cortical model, in which the interior is modeled as cortical bone. The models were compared with published in vivo bite force and bone strain data, and the load paths calculated for each model. The trabecular model, which is preferred because it most closely approximates the actual morphology, was best validated by in vivo data. In all three models, the load path was confined to cortical bone, although its route within the cortex varied depending on the material properties of the inner model. Our analysis shows that most of the force is transferred through the cortical, rather than trabecular bone, and highlights the potential of load path analysis for understanding form-function relationships in the skeleton.

Spatiotemporal partitioning between the endangered San Joaquin kit fox and a novel mesocarnivore community in the urban environment as revealed by camera traps

Urbanization has led to the rapid formation of novel biotic communities, which via interspecific interactions, may affect niche partitioning dynamics of native, sensitive species. The endangered San Joaquin kit fox (Vulpes macrotis mutica; hereafter ‘kit fox’) is a small mesocarnivore inhabiting urban areas of the southern Central Valley of California alongside other urban mesocarnivores. These other species may pose challenges for kit foxes due to a potential lack of evolved mechanisms of defense or avoidance. To address this notion, we evaluated evidence of temporal and spatial partitioning between the kit fox and its potential predators and competitors in an urban environment. We predicted that spatial and temporal overlap would be lower between kit foxes and their predators than with their competitors. We used camera traps with scent lures at high school and college campuses across the southern San Joaquin Valley to monitor the visitation patterns of urban mesocarnivores from 2020 to 2022. We used generalized linear mixed models and AIC model selection to evaluate spatial relationships between kit foxes and other mesocarnivores and we used temporal overlap coefficients and peak activity analysis to evaluate temporal overlap. Kit foxes exhibited low spatial overlap (negative association) with Virginia opossums (Didelphis virginiana) and high spatial overlap (positive association) with domestic cats (Felis catus), domestic dogs (Canis lupus familiaris), northern raccoons (Procyon lotor), and striped skunks (Mephitis mephitis). Kit foxes exhibited low to moderate levels of temporal overlap with other species, the least of which occurring with potential predators (i.e., coyotes (C. latrans), domestic dogs). Our findings support the temporal partitioning and partially support the spatial partitioning hypothesis as potential mechanisms to promote coexistence among mesocarnivores in an urban environment.

BLOOD LEAD CONCENTRATION ANALYSIS BETWEEN REHABILITATION FACILITY AND PERI-URBAN ENVIRONMENT VIRGINIA OPOSSUMS (DIDELPHIS VIRGINIANA) OF ILLINOIS.

The Virginia opossum (Didelphis virginiana) is the only marsupial in North America with a natural range north of Mexico. Its range is widespread throughout the United States, including Illinois. Virginia opossums are opportunistic omnivores that will eat a variety of fruits, grains, insects, and even carrion. They are a ground-foraging species, with frequent soil contact allowing for potential exposure to heavy metal contaminants in terrestrial environments. They are also opportunists that thrive in human-altered ecosystems, including peri-urban environments. Lead is a naturally occurring element and highly toxic metal that is abundant in the soil, primarily due to anthropogenic factors such as fossil fuel use, paint, industrial production waste, and battery recycling. Furthermore, carrion and offal piles containing lead bullet fragments remaining from hunting result in environmental contamination of this heavy metal. Few studies exist on lead exposure in the Virginia opossum. This pilot study evaluates the blood lead burden of 16 free-ranging Virginia opossums presented to a wildlife rehabilitation facility, compared with 16 wild-caught Virginia opossums trapped in grassland, maintained forest, or mixed ecotones. In other species, lead toxicosis has been associated with cognitive dysfunction that may increase the likelihood of trauma or injury in a free-ranging setting. In addition, a high incidence of Virginia opossums presenting for wildlife rehabilitation do so as result of human-animal conflict. Therefore, it was anticipated that individuals presenting for wildlife rehabilitation would have significantly higher blood lead concentrations than their free-roaming counterparts. In this study, every Virginia opossum had measurable blood lead concentrations and the average blood lead concentration of the rehabilitated group was 2.7 times higher than that measured in wild-caught Virginia opossums. These results suggest that Virginia opossums are exposed and susceptible to lead toxicosis due to their diet and foraging behaviors, making them more prone to subsequent injury and presentation to wildlife rehabilitation centers.

Widespread exposure to anticoagulant rodenticides among common urban mesopredators in Chicago

Anticoagulant rodenticides (ARs) are currently the most common method to control rats in cities, but these compounds also cause morbidity and mortality in non-target wildlife. Little attention has been focused on AR exposure among mesopredators despite their ecological role as scavengers and prey for larger carnivores, thus serving as an important bridge in the biomagnification of rodenticides in food webs. In this study, we sampled liver tissue from raccoons (Procyon lotor; n = 37), skunks (Mephitis mephitis; n = 15), and Virginia opossums (Didelphis virginiana; n = 45) euthanized by pest professionals and brown rats (Rattus norvegicus; n = 101) trapped in alleys in Chicago, USA to evaluate how often these species are exposed to ARs. We tested whether mesopredators had a higher prevalence of ARs and to more AR compounds compared to rats and calculated biomagnification factors (mean concentration in mesopredators/rats) as indicators of biomagnification. Of 93 sampled mesopredators, 100 % were exposed to at least one AR compound, mainly brodifacoum (≥80 %), and 79 % were exposed to multiple AR compounds. We also documented teal stomach contents consistent with the consumption of rat bait and altricial young tested positive to the same AR as their mother, suggesting mammary transfer. Of the 101 rats, 74 % tested positive to at least one AR compound and 32 % were exposed to multiple AR compounds. All mesopredator species had biomagnification factors exceeding 1.00 for brodifacoum (6.57–29.07) and bromadiolone (1.08–4.31). Our results suggest widespread exposure to ARs among urban mesopredators and biomagnification of ARs in mesopredators compared to rats. Policies that limit AR availability to non-target species, such as restricting the sale and use of ARs to licensed professionals in indoor settings, education on alternatives, and more emphasis on waste management may reduce health risks for urban wildlife and people in cities around the world.

Seasonal Variation in Mammalian Mesopredator Spatiotemporal Overlap on a Barrier Island Complex.

Due to lack of apex predators in human-dominated landscapes, mesopredator relationships are complex and spatiotemporal niche partitioning strategies can vary, especially when seasonal shifts in resource availability occur. Our objective was to understand spatiotemporal niche overlap across seasons among mesopredators inhabiting a barrier island complex. We placed 19 unbaited cameras throughout Fort De Soto County Park, Florida, USA between February 2021 and July 2023. Of six mesopredator species detected, three species had >75 detections during both the wet and dry seasons (coyote, Canis latrans; Virginia opossum, Didelphis virginiana; and raccoon, Procyon lotor). Using general linear mixed models, we determined that during the wet season coyote-raccoon and raccoon-opossum detections were positively associated with each other (p < 0.05). During the dry season, raccoon-opossum detections were positively associated, and opossums were more likely to be detected around mangroves. After calculating coefficients of overlap, we found all three species varied their temporal activity between seasons. During the dry season exclusively, all three mesopredators occupied different temporal niches. The park's isolated but developed nature has potentially led to a destabilized mesopredator community. Understanding seasonal mesopredator dynamics of Fort De Soto is particularly important because this park supports a high number of nesting shorebirds and sea turtles, which are known food sources for mesopredators.

Cascading effects of mammal host community composition on tick vector occurrence at the urban human–wildlife interface

AbstractHabitat fragmentation and host community composition are implicated as key drivers of changing tick populations and tick‐borne pathogen dynamics, altering infection risk through coupled socioecological pathways that mediate interactions between tick vectors, vertebrate hosts, and humans. Patterns of host diversity may be particularly idiosyncratic across urbanized landscapes, due to trade‐offs between extreme fragmentation that reduces habitat suitability and access, and human activities that artificially increase resource availability for wildlife. We used camera and hair trap surveys and tick sampling to identify links between landscape composition and configuration, the mammalian host community, and the presence of three tick vector species at a human–wildlife interface in New York City, an emerging area within an endemic region for several tick‐borne diseases. We found that human infrastructures, such as the presence of fences in yards, could affect mammal host community composition by changing the “hardness” of edges between urban greenspaces and residential areas. We identified yard‐ and broader landscape‐level features associated with the presence of urban mammal species, and identified cascading effects of host community composition on tick distribution in yards, suggesting management implications for the mitigation of human exposure to tick‐borne pathogens. In particular, we identified a possible role of ubiquitous mesomammals, such as raccoons (Procyon lotor), in transporting Amblyomma americanum ticks between parks and neighboring residential yards, and confirmed the key role of white‐tailed deer (Odocoileus virginianus) for introducing Ixodes scapularis ticks into yards. Our results challenge assumptions that biodiversity loss in human‐modified areas always increases the risk for tick‐borne diseases. Instead, we found many residential sites had higher mammal species richness and higher detection of low reservoir competent (“dilution”) hosts for Borrelia burgdorferi, such as opossums (Didelphis virginiana), than paired forested greenspaces. Our study highlights the importance of disentangling the mechanisms mediating tick‐borne disease hazard as a critical first step toward reducing urban tick‐borne disease risk.

Widespread exposure to SARS-CoV-2 in wildlife communities

Pervasive SARS-CoV-2 infections in humans have led to multiple transmission events to animals. While SARS-CoV-2 has a potential broad wildlife host range, most documented infections have been in captive animals and a single wildlife species, the white-tailed deer. The full extent of SARS-CoV-2 exposure among wildlife communities and the factors that influence wildlife transmission risk remain unknown. We sampled 23 species of wildlife for SARS-CoV-2 and examined the effects of urbanization and human use on seropositivity. Here, we document positive detections of SARS-CoV-2 RNA in six species, including the deer mouse, Virginia opossum, raccoon, groundhog, Eastern cottontail, and Eastern red bat between May 2022–September 2023 across Virginia and Washington, D.C., USA. In addition, we found that sites with high human activity had three times higher seroprevalence than low human-use areas. We obtained SARS-CoV-2 genomic sequences from nine individuals of six species which were assigned to seven Pango lineages of the Omicron variant. The close match to variants circulating in humans at the time suggests at least seven recent human-to-animal transmission events. Our data support that exposure to SARS-CoV-2 has been widespread in wildlife communities and suggests that areas with high human activity may serve as points of contact for cross-species transmission.

On-host flea phenology and flea-borne pathogen surveillance among mammalian wildlife of the pineywoods of East Texas.

Flea-borne diseases are endemic in Texas, U.S.A., with an increasing incidence of flea-borne typhus and cat scratch disease. Knowledge of flea natural history could provide information to protect public health, yet many knowledge gaps remain outside of plague-endemic regions. Our objective was to characterize seasonal activity patterns of fleas on common mammalian wildlife species and test fleas and wildlife for Rickettsia and Bartonella pathogens. We performed one year of monthly trapping for rodents and medium-sized mammals in a national forest with high recreational use and urban encroachment in East Texas. From 90 mammal captures representing seven species, 101 fleas were collected representing Polygenis spp., Ctenocephalides felis, and Orchopeas species. Virginia opossums (Didelphis virginianus) hosted 99% of the collected fleas (100 fleas) and a single flea was on an eastern woodrat (Neotoma floridana). Flea infestation prevalence of opossums was 79% (23/29). Mean flea abundance was 4.39 fleas, with intensity peaking in spring. One cat flea removed from an opossum was positive for Bartonella henselae. Furthermore, we identified tissue or blood of four raccoons (Procyon lotor) and one golden mouse (Ochrotomys nuttalli) positive for Rickettsia amblyommatis. These findings provide an ecological basis for the maintenance of vectors and pathogens from sylvatic settings.

Serosurveillance and the first detection of Bourbon virus RNA in a wildlife host.

Bourbon virus (BRBV) is an emerging pathogen that can cause severe and fatal disease in humans. BRBV is vectored by Amblyomma americanum (lone star ticks), which are widely distributed across the central, southern, and eastern United States. Wildlife species are potentially important for the maintenance and transmission of BRBV, but little is known about which species are involved, and what other factors play a role in the exposure to BRBV. To assess the exposure risk to BRBV among wildlife in the St. Louis area, we collected sera from 98 individuals, representing 6 different mammalian species from two locations in St. Louis County: Tyson Research Center (TRC) and WildCare Park (WCP) from fall 2021 to spring 2023. The sera were used in a BRBV neutralization assay to detect neutralizing antibodies and RT-qPCR for viral RNA analysis. We also sampled and compared the abundance of A. americanum ticks at the two locations and modeled which factors influenced BRBV seropositivity across species. In TRC, we observed a high rate of seropositivity in raccoons (Procyon lotor, 23/25), and white-tailed deer (Odocoileus virginianus, 18/27), but a low rate in opossums (Didelphis virginiana, 1/18). Neutralizing antibodies were also detected in sampled TRC bobcats (Lynx rufus, 4/4), coyotes (Canis latrans, 3/3), and a red fox (Vulpes vulpes, 1/1). The virological analysis identified BRBV RNA in one of the coyote serum samples. In contrast to TRC, all sera screened from WCP were negative for BRBV-specific neutralizing antibodies, and significantly fewer ticks were collected at WCP (31) compared to TRC (2,316). Collectively, these findings suggest that BRBV circulates in multiple wildlife species in the St. Louis area and that tick density and host community composition may be important factors in BRBV ecology.

Morphological and molecular data on helminths of Didelphis virginiana and Philander vossi (Mammalia: Didelphidae) from the Yucatán Peninsula, southeast Mexico

In the present study, helminths from six Didelphis virginiana and one Philander vossi are reported using morphological techniques (clearing, staining, and scanning electron microscopy). Additionally, the 28S rRNA sequences of individuals from nine helminth taxa are provided. Phylogenetic analyses were performed with the new 28S rRNA sequences to confirm the identification and the genealogical relationships of the parasites. Thirteen helminth taxa were identified, comprising the trematodes Brachylaima sp. and Platynosomum illiciens, the cestode Mathevotaenia sp., the nematodes Cruzia americana, Cruzia tentaculata, Viannaia arriaguensis, Viannaia sp., Travassostrongylus sp., Strongyloides sp., Turgida turgida, Trichuris minuta, and Trichuris sp., and the acanthocephalan Oligacanthorhynchus microcephalus. All opossums were infected with at least four helminth taxa. In total, 17 new 28S rRNA sequences from nine helminth taxa were provided. These data, combined with previous records in Mexico, increase the number of helminth taxa parasitizing D. virginiana and P. vossi to 41 and 29, respectively. However, these reports are incompletes and concentered in localities of some states. It is possible that new surveys in the Nearctic and even Neotropical regions will reveal a higher helminth diversity in these mammals in the country.

Detection of macropodid alphaherpesvirus 2 infection and lesions in sudden death of a captive Virginia opossum and a water opossum.

Macropodid alphaherpesvirus 2 (MaAHV2) is best described in macropods and has been implicated in outbreaks among captive marsupial populations in Australia. Natural disease caused by herpesviruses has not been reported previously in opossum species, to our knowledge. One Virginia opossum (Didelphis virginiana) and 1 water opossum (Chironectes minimus) were submitted for postmortem examination from a zoo that housed 6 opossums, all of which died within several weeks. Red kangaroos (Macropus rufus) and red-necked wallabies (Macropus rufogriseus) were also present at the facility. Liver samples from both opossums were submitted for transmission electron microscopy and whole-genome sequencing. Microscopically, both opossums had multifocal necrosis in the liver and lung, with intranuclear inclusion bodies within hepatocytes and pneumocytes. Another significant finding in the Virginia opossum was sepsis, with isolation of Streptococcus didelphis from various organs. Ultrastructural analysis of formalin-fixed liver tissue identified herpesviral replication complexes in both opossums; negative-stain electron microscopy of unfixed liver tissue repeatedly yielded a negative result. The herpesvirus had >99% nucleotide identity with MaAHV2. These 2 cases indicate that both opossum species are susceptible to MaAHV2 infection, and the outbreak has implications for mixed-species facilities that house macropods.

Raccoons and opossums respond similarly to high and low development in the East Texas Pineywoods

Over the last half-century, cities across the southeastern United States have experienced consistent human population growth, which has increased the rate of urban sprawl and subsequent habitat fragmentation with resource subsidies from development. The northern raccoon (Procyon lotor) and Virginia opossum (Didelphis virginiana) are mesopredators commonly associated with and benefiting from urbanized areas. We conducted a small-scale study of raccoons and opossums within the Nacogdoches, Texas, in January 2014. We captured and marked individual raccoons and opossums and used occupancy and spatial-capture-recapture models to examine habitat use, detection, and population density in high- and low-density urban study areas. We predicted to find high densities of each mesopredator, relative to other studies in larger metropolitan areas. We also expected to find a response to landscape features in the high-development study area to be closely associated with anthropogenic features and proximity to water, but less pronounced in the low-development study area. Contrary to our predictions, the detectability of each species in the high-development study area had a positive association with areas further from buildings, and raccoon habitat use had a positive association farther from natural water. Similarly, in the low-development study area, we found raccoon detectability had a positive association with areas farther from the urban edge and opossums had a positive association with areas with smaller forest patches (i.e., edge habitat). We were able to estimate 13.66 raccoons/km2 (95% CI 7.10–26.30) in the high-development study area and 12.55 opossums/km2 (95% CI 04.36–36.15) in the low-development study area. Our results indicate high densities of each species, and the first urban density of Virginia opossum using spatial-capture-recapture methods which further supports that these mesopredators are adapted to and benefit from urban-derived resources in urban areas. This study highlights the adaptability of these small mesopredators and the need for managers to consider adaptive strategies across different urban intensities as the ecology of these species may differ as compared to larger cities.

Potential zoonotic role of the tick Amblyomma cf. oblongoguttatum (Ixodida: Ixodidae) in the bacterial transmission of Ehrlichia chaffeensis (Rickettsiales: Anaplasmataceae) in a deciduous tropical forest in Mexico.

Ehrlichia chaffeensis is a bacterium belonging to the Anaplasmataceae family. In Mexico, only 2 species have been recorded in association with tick species and humans. The objective of the present study was to detect the presence of bacteria of the genus Ehrlichia in ticks collected from the Chamela-Cuixmala Biosphere Reserve, Jalisco, Mexico. The collected ticks were identified and analyzed individually by polymerase chain reaction to amplify a fragment of the Anaplasmataceae 16S rRNA gene and the Ehrlichia-specific dsb gene. A total of 204 ticks, corresponding to 5 species of Ixodidae and 1 of Argasidae, were collected from 147 mammals of 6 species and 4 orders; 57 ticks collected from vegetation were also included. Among the total ticks collected, 1.47% (3/204) was positive for Ehrlichia sp. DNA was obtained using the primers EHR 16SD and EHR 16SR for 16S rRNA and DSB-330 and DSB-728 for dsb. The positive samples corresponded to a larva (Amblyomma sp.) associated with Didelphis virginiana and 2 nymphs (Amblyomma cf. oblongoguttatum) infesting Nasua narica. None of the ticks collected from the vegetation tested positive for Ehrlichia sp. DNA on the basis of the 16S rRNA and dsb genes. The sequences from the larvae of Amblyomma sp. and the nymphs of A. cf. oblongoguttatum were similar to those of E. chaffeensis. The phylogenetic analysis inferred with maximum likelihood corroborated the identity as E. chaffeensis. Although the role of these tick species as vectors of E. chaffeensis is still undetermined, the presence of infected ticks in the area indicates a potential zoonotic risk.

Otterly diverse - A high diversity of Dracunculus species (Spirurida: Dracunculoidea) in North American river otters (Lontra canadensis)

The genus Dracunculus contains numerous species of subcutaneous parasites of mammals and reptiles. In North America, there are at least three mammal-infecting species of Dracunculus. Reports of Dracunculus infections have been reported from river otters (Lontra canadensis) since the early 1900s; however, little is known about the species infecting otters or their ecology. Most reports of Dracunculus do not have a definitive species identified because females, the most common sex found due to their larger size and location in the extremities of the host, lack distinguishing morphological characteristics, and few studies have used molecular methods to confirm identifications. Thus, outside of Ontario, Canada, where both D. insignis and D. lutrae have been confirmed in otters, the species of Dracunculus in river otters is unknown. In the current study, molecular characterization of nematodes from river otters revealed a high diversity of Dracunculus species. In addition to confirming D. insignis infections, two new clades were detected. One clade was a novel species in any host and the other was a clade previously detected in Virginia opossums (Didelphis virginiana) from the USA and a domestic dog from Spain. No infections with D. lutrae were detected and neither new lineage was genetically similar to D. jaguape, which was recently described from a neotropical otter (Lontra longicaudis) from Argentina. These data also indicate that Dracunculus spp. infections in otters are widespread throughout Eastern North America. Currently the life cycles for most of the Dracunculus spp. infecting otters are unknown. Studies on the diversity, life cycle, and natural history of Dracunculidae parasites in wildlife are important because the related parasite, D. medinensis (human Guinea worm) is the subject of an international eradication campaign and there are increasing reports of these parasites in new geographic locations and new hosts, including new species in humans and domestic dogs.

Ontology-based taxonomical analysis of experimentally verified natural and laboratory human coronavirus hosts and its implication for COVID-19 virus origination and transmission.

To fully understand COVID-19, it is critical to study all possible hosts of SARS-CoV-2 (the pathogen of COVID-19). In this work, we collected, annotated, and performed ontology-based taxonomical analysis of all the reported and verified hosts for all human coronaviruses including SARS-CoV, MERS-CoV, SARS-CoV-2, HCoV-229E, HCoV-NL63, HCoV-OC43, and HCoV-HKU1. A total of 37 natural hosts and 19 laboratory animal hosts of human coronaviruses were identified based on experimental evidence. Our analysis found that all the verified susceptible natural and laboratory animals belong to therian mammals. Specifically, these 37 natural therian hosts include one wildlife marsupial mammal (i.e., Virginia opossum) and 36 Eutheria mammals (a.k.a. placental mammals). The 19 laboratory animal hosts are also classified as therian mammals. The mouse models with genetically modified human ACE2 or DPP4 were more susceptible to virulent human coronaviruses with clear symptoms, suggesting the critical role of ACE2 and DPP4 to coronavirus virulence. Coronaviruses became more virulent and adaptive in the mouse hosts after a series of viral passages in the mice, providing clue to the possible coronavirus origination. The Huanan Seafood Wholesale Market animals identified early in the COVID-19 outbreak were also systematically analyzed as possible COVID-19 hosts. To support knowledge standardization and query, the annotated host knowledge was modeled and represented in the Coronavirus Infectious Disease Ontology (CIDO). Based on our and others' findings, we further propose a MOVIE model (i.e., Multiple-Organism viral Variations and Immune Evasion) to address how viral variations in therian animal hosts and the host immune evasion might have led to dynamic COVID-19 pandemic outcomes.

Venipuncture and Blood Collection on Conscious Virginia Opossums (Didelphis virginiana).

We developed a venipuncture technique of the ventral caudal vein in conscious Virginia opossums (Didelphis virginiana) in cage traps, enabling blood sampling without anesthesia. Blood samples were successfully collected from all 28 opossums in this study by this technique. Draw volume of <0.1 mL occurred in only four opossums (14%).

Oral Rabies Vaccination of Raccoons (Procyon lotor) across a Development Intensity Gradient in Burlington, Vermont, USA, 2015-2017.

Management of the raccoon rabies virus variant in North America is conducted primarily using oral rabies vaccination (ORV). When a sufficient proportion of the population is vaccinated (∼60%), rabies transmission can be eliminated. To date, ORV programs have successfully controlled and eliminated raccoon rabies in rural areas, but there has been less success in urban areas. We studied the proportions of rabies virus neutralizing antibodies (RVNA) in a raccoon (Procyon lotor) population during a 3-yr ORV trial in developed areas of Burlington, Vermont, US. We used a modified N-mixture model to estimate raccoon abundance, RVNA seroprevalence, and capture rates jointly to examine factors that relate to ORV success to better inform management. We found that raccoon abundance was lower in less-developed areas compared to urban centers. Raccoon RVNA seroprevalence decreased as population abundance increased; it increased as the average age of the population increased. Nontarget opossum (Didelphis virginiana) captures correlated with a decrease in raccoon RVNA seroprevalence in low-development areas, suggesting that they may be competing for baits. The target bait density across the entire study area was 150 baits/km2, but a hand baiting strategy was heavily concentrated on roads, resulting in uneven bait densities within sampling sites (0-484 baits/km2). Uneven bait distribution across the study area may explain low RVNA seroprevalence in some locations. Our results suggest that increases in bait density across the study area may improve RVNA seroprevalence and support annual ORV to account for raccoon population turnover.

Does daily activity overlap of seven mesocarnivores vary based on human development?

Many species of wildlife alter their daily activity patterns in response to co-occurring species as well as the surrounding environment. Often smaller or subordinate species alter their activity patterns to avoid being active at the same time as larger, dominant species to avoid agonistic interactions. Human development can complicate interspecies interactions, as not all wildlife respond to human activity in the same manner. While some species may change the timing of their activity to avoid being active when humans are, others may be unaffected or may benefit from being active at the same time as humans to reduce predation risk or competition. To further explore these patterns, we used data from a coordinated national camera-trapping program (Snapshot USA) to explore how the activity patterns and temporal activity overlap of a suite of seven widely co-occurring mammalian mesocarnivores varied along a gradient of human development. Our focal species ranged in size from the large and often dominant coyote (Canis latrans) to the much smaller and subordinate Virginia opossum (Didelphis virginiana). Some species changed their activity based on surrounding human development. Coyotes were most active at night in areas of high and medium human development. Red fox (Vulpes vulpes) were more active at dusk in areas of high development relative to areas of low or medium development. However, because most species were primarily nocturnal regardless of human development, temporal activity overlap was high between all species. Only opossum and raccoon (Procyon lotor) showed changes in activity overlap with high overlap in areas of low development compared to areas of moderate development. Although we found that coyotes and red fox altered their activity patterns in response to human development, our results showed that competitive and predatory pressures between these seven widespread generalist species were insufficient to cause them to substantially alter their activity patterns.