Blood analytes of clinically normal and diseased neonatal and weaned farmed white-tailed deer (Odocoileus virginianus) fawns.

Recent research focused on farmed deer has exposed many knowledge gaps regarding health assessment protocols for white-tailed deer (WTD). The objectives of this study were to establish de novo blood analyte reference intervals for farmed WTD fawns at birth (1-2 days of age; n = 84) and again at weaning (76-125 days of age; n = 28), to compare data at birth and at weaning to understand how these analytes are affected by the intrinsic factors age and sex in clinically normal WTD fawns, and to compare between clinically normal and sick WTD weanlings (respiratory disease n = 12; orbivirus-infected n = 6). Reference intervals were established for WTD fawns at birth and weaning. Female WTD neonates had significantly higher red blood cell counts, hematocrit, and hemoglobin compared to males. Most blood analytes were significantly different in clinically normal WTD neonates compared to weanlings, suggesting an effect of age. The observed sex- and age-related variations in WTD highlight the need to establish reference intervals that account for intrinsic factors. The comparison of clinically normal and sick WTD weanlings in this study identified higher MCHC and absolute monocytes in sick weanlings but these findings were presumably not biologically relevant given the small sample size for sick fawns. While the reference interval data presented herein will be useful for the veterinary care of WTD fawns at critical time periods in a high-density farm setting, this study also demonstrates the need to identify more sensitive and specific biomarkers for the assessment of health status in farmed WTD with specific underlying diseases.

Distribution of Baylisascaris procyonis in Raccoons (Procyon lotor) in Florida, USA.

Baylisascaris procyonis, or raccoon roundworm, is an intestinal nematode parasite of raccoons (Procyon lotor) that is important to public and wildlife health. Historically, the parasite was uncommon in the southeastern US; however, the range of B. procyonis has expanded to include Florida, US. From 2010 to 2016, we opportunistically sampled 1,030 raccoons statewide. The overall prevalence was 3.7% (95% confidence interval=2.5-4.8%) of sampled individuals, and infection intensity ranged from 1 to 48 (mean±standard deviation 9.9±4.0). We found raccoon roundworm in 9/56 (16%) counties sampled, and the percent positive ranged from 1.1% to 13.3% of specimens collected per county. Including previously published data, B. procyonis was detected in 11 Florida counties. We used logistic regression to estimate the contribution of raccoon demographic variables and the presence of the endoparasite Macracanthorhynchus ingens to B. procyonis detection in Florida. Following the model selection process we found housing density, M. ingens presence, and urbanicity to be predictive of raccoon roundworm presence. We also found substantial among-county variation. Raccoon sex and age were not useful predictors. Public health officials, wildlife rehabilitators, wildlife managers, and others should consider any Florida raccoon to be potentially infected with B. procyonis, particularly in areas where housing density is high.

PSEUDORABIES (AUJESZKY'S DISEASE) IS AN UNDERDIAGNOSED CAUSE OF DEATH IN THE FLORIDA PANTHER (PUMA CONCOLOR CORYI).

Feral swine (Sus scrofa), an important prey species for the endangered Florida panther (Puma concolor coryi), is the natural host for pseudorabies virus (PRV). Prior to this study, PRV had been detected in just three panthers. To determine the effect of PRV on the panther population, we prospectively necropsied 199 panthers and retrospectively reviewed necropsy and laboratory findings, reexamined histology, and tested archived tissues using real-time PCR from 46 undiagnosed panther mortalities. Seven additional infections (two prospective, five retrospective) were detected for a total of 10 confirmed panther mortalities due to PRV. To further evaluate the effect of PRV, we categorized radio-collared (n=168) and uncollared panther mortalities (n=367) sampled from 1981 to 2018 based on the likelihood of PRV infection as confirmed, probable, suspected, possible, or unlikely/negative. Of 168 radio-collared panthers necropsied, PRV was the cause of death for between eight (confirmed; 4.8%) and 32 (combined confirmed, probable, suspected, and possible categories; 19.0%) panthers. The number of radio-collared panther mortalities due to PRV was estimated to be 15 (95% empirical limits: 12-19), representing 8.9% (confidence interval: 4.6-13.2%) of mortalities. Gross necropsy findings in 10 confirmed cases were nonspecific. Microscopic changes included slight to mild perivascular cuffing and gliosis (primarily in the brain stem), lymphoplasmacytic meningoencephalitis (cerebral cortex), and intranuclear inclusion bodies (adrenal medulla). The PRV glycoprotein C gene sequences from three positive panthers grouped with the sequence from a Florida feral swine. Our findings indicate that PRV may be an important and underdiagnosed cause of death in Florida panthers.

Modeling Abundance of Culicoides stellifer, a Candidate Orbivirus Vector, Indicates Nonrandom Hemorrhagic Disease Risk for White-Tailed Deer (Odocoileus virginianus).

(1) Background: Hemorrhagic diseases in white-tailed deer (Odocoileus virginianus) are caused by orbiviruses and have significant economic impact on the deer ranching industry in the United States. Culicoides stellifer is a suspected vector of epizootic hemorrhagic disease virus (EHDV), with recent field evidence from Florida, but its natural history is poorly understood. Studying the distribution and abundance of C. stellifer across the landscape can inform our knowledge of how virus transmission can occur locally. We may then target vector management strategies in areas where viral transmission can occur. (2) Methods: Here, we used an occupancy modeling approach to estimate abundance of adult C. stellifer females at various physiological states to determine habitat preferences. We then mapped midge abundance during the orbiviral disease transmission period (May-October) in Florida. (3) Results: We found that overall, midge abundance was positively associated with sites in closer proximity to large-animal feeders. Additionally, midges generally preferred mixed bottomland hardwood and agricultural/sand/water habitats. Female C. stellifer with different physiological states preferred different habitats. (4) Conclusions: The differences in habitat preferences between midges across states indicate that disease risk for deer is heterogeneous across this landscape. This can inform how effective vector management strategies should be implemented.

Three New Orbivirus Species Isolated from Farmed White-Tailed Deer (Odocoileus virginianus) in the United States.

We report the detection and gene coding sequences of three novel Orbivirus species found in six dead farmed white-tailed deer in the United States. Phylogenetic analyses indicate that the new orbiviruses are genetically closely related to the Guangxi, Mobuck, Peruvian horse sickness, and Yunnan orbiviruses, which are thought to be solely borne by mosquitos. However, four of the six viruses analyzed in this work were found as co-infecting agents along with a known cervid pathogen, epizootic hemorrhagic disease virus-2 (EHDV-2), raising questions as to whether the new viruses are primary pathogens or secondary pathogens that exacerbate EHDV-2 infections. Moreover, EHDV-2 is known to be a Culicoides-borne virus, raising additional questions as to whether Culicoides species can also serve as vectors for the novel orbiviruses, if mosquitoes can vector EHDV-2, or whether the deer were infected through separate bites by the insects. Our findings expand knowledge of the possible viral pathogens of deer in the United States. Moreover, due to the close genetic relatedness of the three new orbiviruses to viruses that are primary pathogens of cattle and horses, our findings also underscore a crucial need for additional research on the potential role of the three new orbiviruses as pathogens of other animals.

Multi-scale patterns of tick occupancy and abundance across an agricultural landscape in southern Africa.

Land use influences the prevalence and distribution of ticks due to the intimate relationship of ticks with their environment. This relationship occurs because land use alters two essential tick requirements: vertebrate hosts for blood meals and a suitable microclimate when off-host. Given the risks to human and animal health associated with pathogens transmitted by ticks, there is an ongoing need to understand the impact of environmental drivers on tick distributions. Here, we assessed how landscape features, neighborhood effects, and edges influenced tick occupancy and abundance across an agricultural landscape in southern Africa. We found that Rhipicephalus appendiculatus and Rhipicephalus simus increased in abundance closer to protected savanna, while Haemaphysalis elliptica increased in abundance closer to human habitation. The composition of the landscape surrounding savanna patches also differentially influenced the occupancy of each tick species; H. elliptica was more likely to be found in savanna patches surrounded by subsistence agriculture while R. appendiculatus and R. simus were more likely to be found in savanna surrounded by sugarcane monocultures. At the local scale we found that R. appendiculatus and R. simus avoided savanna edges. The availability of hosts and variation in vegetation structure between commercial agriculture, subsistence agriculture, and savanna likely drove the distribution of ticks at the landscape scale. Understanding how anthropogenic land use influences where ticks occur is useful for land use planning and for assessing public and animal health risks associated with ticks and tick-borne diseases.

Molecular characterization of a novel reassortment Mammalian orthoreovirus type 2 isolated from a Florida white-tailed deer fawn.

Mammalian orthoreovirus (MRV) is the type species of the genus Orthoreovirus and causes a range of significant respiratory, nervous or enteric diseases in humans and animals. In 2016 a farmed white-tailed deer (Odocoileus virginianus) fawn became ill, displaying clinical signs of lethargy, dehydration, and profuse foul-smelling diarrhea. A necropsy was performed after the three-week-old fawn died and various tissue samples were submitted to the University of Florida's Cervidae Health Research Initiative for diagnostic evaluation. Aliquots of homogenized heart, liver, and spleen tissues were inoculated onto Vero E6 cells. After virus-specific cytopathic effects (CPE) were detected in Vero cells inoculated with spleen homogenate, infected cells were fixed in glutaraldehyde and analyzed by transmission electron microscopy (TEM), which revealed icosahedral virus particles approximately 75 nm in diameter with morphologies consistent with those of reoviruses within the cytoplasm of the infected cells. RNA extracted from virions in the spent media of infected cells with advanced CPE was used to prepare a cDNA library, which was sequenced using an Illumina MiSeq sequencer. Complete coding sequences for ten separate reovirus segments were attained, and these indicated the isolated agent was a MRV. Genetic and phylogenetic analyses based on the outer capsid sigma-1 (σ1) protein gene sequences supported the Florida white-tailed fawn isolate as a type 2 MRV that branched as the sister group to a MRV-2 strain previously characterized from the urine of a moribund lion (Panthera leo) in Japan. However, analyses based on 7/10 genes (L1-L2, M2-M3, S2-S4) supported the white-tailed deer MRV as the closest relative to a type 3 MRV strain isolated from a dead mink in China. These data suggest the white-tailed deer MRV may have resulted from the natural reassortment of MRVs originating from multiple wildlife species. To our knowledge, this is the first detection of MRV-2 infection in a white-tailed deer. Continued surveillance efforts are needed to determine whether this MRV-2 strain poses a health threat to farmed white-tailed deer populations.

Field data implicating Culicoides stellifer and Culicoides venustus (Diptera: Ceratopogonidae) as vectors of epizootic hemorrhagic disease virus.

BACKGROUND: Epizootic hemorrhagic disease virus (EHDV) is an Orbivirus of veterinary importance which is transmitted by biting midges of the genus Culicoides (Diptera: Ceratopogonidae) to ruminants. Culicoides sonorensis Wirth & Jones, the only confirmed vector of EHDV in the USA, is rare in the southeastern states where transmission persists, suggesting that other Culicoides species transmit EHDV in this region. The present study aimed to determine which Culicoides species transmitted EHDV in Florida and Alabama, two states in the southeastern USA. Viral RNA was detected in field-collected midges using molecular methods. These data are presented alongside data on Culicoides blood meal analysis, white-tailed deer (Odocoileus virginianus) aspiration, and seasonality to demonstrate an interaction between potential vector species and EHDV hosts. RESULTS: Out of 661 pools tested, 20 pools were positive for EHDV viral RNA, including six pools from Culicoides stellifer (Coquillett) and 14 pools from Culicoides venustus Hoffman. The overall infection rate was 0.06% for C. stellifer and 2.18% for C. venustus. No positive pools were identified for a further 17 species. Serotypes identified in Culicoides included EHDV-2, EHDV-6, and coinfections of EHDV-2 and EHDV-6 and were identified in similar proportions to serotypes in deer at 3 of 4 deer farms. Viral detections conducted in Alabama also identified one positive pool of C. venustus. Blood meal analysis revealed that both Culicoides species fed on white-tailed deer (verified through aspiration), fallow deer, and elk, species for which EHDV viremia has been documented. Seasonality data indicated that both species were present throughout the period in which viral transmission occurred to EHDV hosts in 2016 in addition to the 2017 epizootic. CONCLUSIONS: Our finding of EHDV positive pools of field-collected C. stellifer and C. venustus and an interaction between these species and EHDV hosts satisfy two of the four criteria for vector incrimination as set by the World Health Organization. Determining the vectors of EHDV is an important step towards developing sound strategies for the control of vector Culicoides and management of EHDV in the southeastern USA.

Epidemiology of Bluetongue Virus and Epizootic Hemorrhagic Disease Virus in Beef Cattle on a Ranch in South-Central Florida.

Bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) infect a variety of wild and domestic ruminant hosts in the United States, with outcomes ranging from subclinical infection to clinical disease resulting in mortality. Because cattle have been suggested as a temporary reservoir for both BTV and EHDV, ongoing national surveillance for these viruses may benefit from inclusion of domestic cattle as a supplement to current programs, such as surveillance of wild white-tailed deer. To better understand the prevalence of BTV and EHDV in cattle, we surveyed for viral RNA (vRNA) in the blood of 1,604 beef cattle on a south-central Florida cattle ranch over 3 years. While overall prevalence of vRNA in blood was low (<2% for either virus), the occurrence of vRNA was much higher in young animals: in 2016, 24% of animals 2 years old were positive by PCR for either BTV or EHDV. Our results suggest that cattle are a likely temporary reservoir for these viruses in Florida, and could provide additional information on the spatial distribution, viral diversity, and timing of emergence of these viruses, particularly if surveillance was restricted to cattle ≤2 years of age.

Genome Sequences of a Novel Strain of Big Cypress Orbivirus Isolated from a Dead Florida White-Tailed Deer (Odocoileus virginianus).

Here, we report the coding sequences of Big Cypress orbivirus isolated from a dead white-tailed deer (Odocoileus virginianus) from Florida in 2017. To our knowledge, this is the first detection of Big Cypress orbivirus in a vertebrate host.

The impact of vector control on the prevalence of Theileria cervi in farmed Florida white-tailed deer, Odocoileus virginianus.

BACKGROUND: Vector-borne diseases exert a global economic impact to the livestock industry. Understanding how agriculture practices and acaricide usage affect the ecology of these diseases is important for making informed management decisions. Theileria cervi is a hemoprotozoan parasite infecting white-tailed deer (Odocoileus virginianus) and is transmitted by the lone star tick, Amblyomma americanum. The purpose of this study was to determine if acaricide treatment decreased hematozoan prevalence in farmed white-tailed deer when compared to geographically-close wild deer or altered the genotypes of T. cervi present. RESULTS: We compared prevalence of T. cervi in 52 farmed adult white-tailed deer which were regularly treated with permethrin and ivermectin, 53 farmed neonates that did not receive treatment for vector control, and 42 wild deer that received no form of chemical vector control. Wild deer had significantly higher prevalence of T. cervi than farmed deer. Additionally, no neonate fawns tested positive for T. cervi, and we found that age was a significant predictor of infection status. We found no difference in genotypic variation in T. cervi isolates between adjacent herds of farmed and wild white-tailed deer, although a divergent genotype X was identified. Chronic infection with T. cervi had no significant effects on mortality in the white-tailed deer. CONCLUSIONS: We found significantly lower prevalence of T. cervi infection in farmed (40%) compared to wild white-tailed deer (98%), which may be due to the inclusion of chemical vector control strategies. More work is needed to determine the implications, if any, of mixed genotypic infections of T. cervi, although we found no significant effect of infection with Theileria on mortality in farmed deer. Theileria infection does sometimes cause disease when an animal is stressed, immunosuppressed, or translocated from non-endemic to endemic regions.

Complete Genome Sequence of Mobuck Virus Isolated from a Florida White-Tailed Deer (Odocoileus virginianus).

Here, we report the complete genome sequence of mobuck virus isolated from a Florida white-tailed deer (Odocoileus virginianus) in 2017. This is the second report of mobuck virus in the United States and expands the known geographic range of this novel orbivirus into Florida.

PREVALENCE OF PARELAPHOSTRONGYLUS ANDERSONI IN WHITE-TAILED DEER, OTHER CERVIDS, AND BOVIDS IN NORTHERN FLORIDA.

Parelaphostrongylus andersoni, the muscleworm, commonly infects white-tailed deer (Odocoileus virginianus) and also infects caribou (Rangifer tarandus [R.t.] groenlanicus, R.t. grantii, R.t. tarandus, and R.t. caribou). Heavy infection with P. andersoni leads to weakness in the hindquarters, abnormal gait, and pulmonary lesions. The geographical range and full host spectrum of this parasite are not fully known. This study aims to understand host specificity better, especially in nonnative cervids and bovids. This study involved the collection of 140 fecal samples from native and nonnative cervid and bovid species, and 34 snail specimens. With the use of real-time PCR, we found 4/47 (8.5%) O. virginianus fecal samples were positive for P. andersoni. No previously undocumented species of cervids or bovids were found to be infected. Further research is warranted to understand P. andersoni range, host distribution, and potential impact on host health.

Characterization of mule deerpox virus in Florida white-tailed deer fawns expands the known host and geographic range of this emerging pathogen.

Infections caused by mule deerpox virus (MDPV) have been sporadically reported in North American cervids. White-tailed deer (Odocoileus virginianus) fawns from a farm located in South Central Florida presented with ulcerative and crusting lesions on the coronary band as well as the mucocutaneous tissues of the head. Evaluation of the crusted skin lesions was undertaken using microscopic pathology and molecular techniques. A crusted skin sample was processed for virus isolation in four mammalian cell lines. The resulting isolate was characterized by negative staining electron microscopy and deep sequencing. Histopathologic evaluation of the skin lesions from the fawns revealed a hyperplastic and proliferative epidermis with ballooning degeneration of epidermal and follicular keratinocytes with intracytoplasmic eosinophilic inclusions. Electron microscopy of cell culture supernatant demonstrated numerous large brick-shaped particles typical of most poxviruses. Polymerase chain reaction assays followed by Sanger sequencing revealed a poxvirus gene sequence nearly identical to that of previous strains of MDPV. The full genome was recovered by deep sequencing and genetic analyses supported the Florida white-tailed deer isolate (MDPV-F) as a strain of MDPV. Herein, we report the first genome sequence of MDPV from a farmed white-tailed deer fawn in the South Central Florida, expanding the number of locations and geographic range in which MDPV has been identified.

Estimating the Geographic Distribution of Host-Seeking Adult Amblyomma americanum (Acari: Ixodidae) in Florida.

The lone star tick, Amblyomma americanum, is the most commonly reported human-biting tick in the southeastern United States and is a vector for several human and livestock pathogens. Although it is endemic to Florida, little is known about the ecological preferences and current spatial distribution within the state. Using occurrence records of adult A. americanum collected between August 2015 and September 2016, a logistic regression model was used to estimate environmental associations, as well as to predict the distribution of the tick at a one hectare resolution. Occurrence of adult lone star ticks was associated with land cover and bioclimatic variables, namely the presence of forested areas and precipitation seasonality. The estimated spatial distribution indicated that central and northern regions show greater suitability than the southern half of the state. Furthermore, areas predicted to be suitable for the species decreases from north to south with very little area deemed suitable in the far southern reaches of the state. High heterogeneity in the distribution of suitable habitat has implications for the distribution of tick-borne disease cases in the state. The subcounty resolution of the estimated distribution is an improvement over distributions currently published and may better inform the public and state or federal agencies of potential risk of exposure to A. americanum and its associated pathogens.

Wild pigs as sentinels for hard ticks: A case study from south-central Florida.

As a result of shifts in the habitable range of ticks due to climate change and the ongoing threat of exotic tick species introductions, efficient surveillance tools for these pests and disease vectors are needed. Wild pigs are habitat generalists, distributed throughout most of the United States, and often hunted recreationally or removed as part of management programs, making them potentially useful sentinel hosts for ticks. We compared ticks collected from captured wild pigs and standard tick dragging methods on a south-central Florida cattle ranch from May 2015-August 2017. Three hundred and sixteen wild pigs were surveyed, and 84 km spanning three habitat types (seminative pasture, improved pasture, and hammock) were dragged. In total, 1023 adults of four species (Amblyomma auricularium, Amblyomma maculatum, Dermacentor variabilis, and Ixodes scapularis) were collected from wild pigs, while 39 adults of three species (A. auricularium, A. maculatum, and I. scapularis) were collected from drags. Only one immature specimen, a nymph, was collected from a pig, while dragging collected 2808 larvae and 150 nymphs. Amblyomma maculatum comprised 96% of adults collected from pigs, while A. maculatum, I. scapularis, and A. auricularium comprised 38%, 33%, and 28% of adults collected from drags, respectively. Adults of all tick species found on drags were found on pigs, and wild pig surveillance detected adults of an additional species not found on drags. Dragging was far superior for collection of immatures but not for adults of most species found in this study. These findings suggest wild pigs could be used as a sentinel for the detection of tick species. When combined with ongoing wild pig research, hunting, or management, wild pig surveillance can provide an effective method to survey for adult tick presence of some species of interest and may assist in tracking the range expansion of some tick species.

Complete Genome Sequence of Epizootic hemorrhagic disease virus Serotype 6, Isolated from Florida White-Tailed Deer (Odocoileus virginianus).

Here, we report the complete genome sequence of Epizootic hemorrhagic disease virus (EHDV) serotype 6 (EHDV-6), isolated from a Florida white-tailed deer (Odocoileus virginianus) in 2016. To our knowledge, this is the first full genome sequence determined for an EHDV-6 isolate from Florida.

Natural History of Plasmodium odocoilei Malaria Infection in Farmed White-Tailed Deer.

White-tailed deer (Odocoileus virginianus), an ecologically and economically important species, are the most widely distributed large animals in North America. A recent study indicated that up to 25% of all white-tailed deer may be infected with Plasmodium odocoilei, a malaria parasite belonging to the distinct clade of ungulate-infecting Plasmodium spp. Because the clinical impact of P. odocoilei on deer health and survival is unknown, we undertook a retrospective longitudinal study of farmed Floridian O. virginianus fawns. We found that a substantial proportion (21%) of fawns acquire malaria infection during the first 8 months of life. Some animals naturally clear P. odocoilei infection, while other animals remain persistently positive. Importantly, we found that animals that acquire malaria parasites very early in life have poor survival compared to animals that remain uninfected. Our report thus provides the first evidence of a clinically significant impact of malaria infection in young deer.IMPORTANCE Malaria parasites of the genus Plasmodium are known to infect a variety of vertebrate hosts, including ungulates (hoofed mammals). A recent study found that up to a quarter of white-tailed deer (Odocoileus virginianus) in North America are infected with the parasite Plasmodium odocoilei In addition to occupying an important ecological niche, white-tailed deer are popular game animals and deer farming represents a rapidly growing industry. However, the effect of P. odocoilei infection in this ecologically and economically important ungulate species is unknown. Our work is significant because (i) we identified a high prevalence of P. odocoilei in farmed deer and (ii) we found evidence for both cleared and persistent infection, as well as an association with decreased survival of young fawns.

Macacine Herpesvirus 1 Antibody Prevalence and DNA Shedding among Invasive Rhesus Macaques, Silver Springs State Park, Florida, USA.

We compiled records on macacine herpesvirus 1 (McHV-1) seroprevalence and, during 2015-2016, collected saliva and fecal samples from the free-ranging rhesus macaques of Silver Springs State Park, a popular public park in central Florida, USA, to determine viral DNA shedding and perform sequencing. Phylogenetic analysis of the US5 and US5-US6 intragenic sequence from free-ranging and laboratory McHV-1 variants did not reveal genomic differences. In animals captured during 2000-2012, average annual seroprevalence was 25% ± 9 (mean ± SD). We found 4%-14% (95% CI 2%-29%) of macaques passively sampled during the fall 2015 mating season shed McHV-1 DNA orally. We did not observe viral shedding during the spring or summer or from fecal samples. We conclude that these macaques can shed McHV-1, putting humans at risk for exposure to this potentially fatal pathogen. Management plans should be put in place to limit transmission of McHV-1 from these macaques.

EVIDENCE OF PSEUDORABIES VIRUS SHEDDING IN FERAL SWINE ( SUS SCROFA) POPULATIONS OF FLORIDA, USA.

: Feral swine ( Sus scrofa) are a pathogen reservoir for pseudorabies virus (PrV). The virus can be fatal to wildlife and contributes to economic losses in the swine industry worldwide. National surveillance efforts in the US use serology to detect PrV-specific antibodies in feral swine populations, but PrV exposure is not a direct indicator of pathogen transmission among conspecifics or to non-suid wildlife species. We measured antibody production and the presence of PrV DNA in four tissue types from feral swine populations of Florida, US. We sampled blood, nasal, oral, and genital swabs from 551 individuals at 39 sites during 2014-16. Of the animals tested for antibody production, 224 of 436 (51%) feral swine were antibody positive while 38 of 549 feral swine (7%) tested for viral shedding were quantitative polymerase chain reaction (qPCR)-positive for PrV. The detection of PrV DNA across all the collected sample types (blood, nasal, oral, and genital [vaginal] swabs) suggested viral shedding via direct (oronasal or venereal), and potentially indirect (through carcass consumption), routes of transmission among infected and susceptible animals. Fourteen of 212 seronegative feral swine were qPCR-positive, indicating 7% false negatives in the serologic assay. Our findings suggest that serology may underestimate the actual infection risk posed by feral swine to other species and that feral swine populations in Florida are capable of shedding the virus through multiple routes.