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We investigated the landscape epidemiology of a globally distributed mammal, the wild pig (Sus scrofa), in Florida (U.S.), where it is considered an invasive species and reservoir to pathogens that impact the health of people, domestic animals, and wildlife. Specifically, we tested the hypothesis that two commonly cited factors in disease transmission, connectivity among populations and abundant resources, would increase the likelihood of exposure to both pseudorabies virus (PrV) and Brucella spp. (bacterial agent of brucellosis) in wild pigs across the Kissimmee Valley of Florida. Using DNA from 348 wild pigs and sera from 320 individuals at 24 sites, we employed population genetic techniques to infer individual dispersal, and an Akaike information criterion framework to compare candidate logistic regression models that incorporated both dispersal and land cover composition. Our findings suggested that recent dispersal conferred higher odds of exposure to PrV, but not Brucella spp., among wild pigs throughout the Kissimmee Valley region. Odds of exposure also increased in association with agriculture and open canopy pine, prairie, and scrub habitats, likely because of highly localized resources within those land cover types. Because the effect of open canopy on PrV exposure reversed when agricultural cover was available, we suggest that small-scale resource distribution may be more important than overall resource abundance. Our results underscore the importance of studying and managing disease dynamics through multiple processes and spatial scales, particularly for non-native pathogens that threaten wildlife conservation, economy, and public health.

Rationale/background: Though much smaller than the bovine industry, the porcine sector in Argentina involves a large number of farms and represents a significant economic sector. In recent years Argentina has implemented a national registry of swine movements amongst other measures, in an effort to control and eventually eradicate endemic Aujesky's disease. Such information can prove valuable in assessing the risk of transmission between farms for endemic diseases but also for other diseases at risk of emergence., Methods: Shipment data from 2011 to 2016 were analyzed in an effort to define strategic locations and times at which control and surveillance efforts should be focused to provide cost-effective interventions. Social network analysis (SNA) was used to characterize the network as a whole and at the individual farm and market level to help identify important nodes. Spatio-temporal trends of pig movements were also analyzed. Finally, in an attempt to classify farms and markets in different groups based on their SNA metrics, we used factor analysis for mixed data (FAMD) and hierarchical clustering., Results: The network involved approximate 136,000 shipments for a total of 6 million pigs. Over 350 markets and 17,800 production units participated in shipments with another 83,500 not participating. Temporal data of shipments and network metrics showed peaks in shipments in September and October. Most shipments where within provinces, with Buenos Aires, Cordoba and Santa Fe concentrating 61% of shipments. Network analysis showed that markets are involved in relatively few shipments but hold strategic positions with much higher betweenness compared to farms. Hierarchical clustering yielded four groups based on SNA metrics and node characteristics which can be broadly described as: 1. small and backyard farms; 2. industrial farms; 3. markets; and 4. a single outlying market with extreme centrality values., Conclusion: Characterizing the network structure and spatio-temporal characteristics of Argentine swine shipments provides valuable information that can guide targeted and more cost-effective surveillance and control programs. We located key nodes where efforts should be prioritized. Pig network characteristics and patterns can be used to create dynamic disease transmission models, which can both be used in assessing the impact of emerging diseases and guiding efforts to eradicate endemic ones., Competing Interests: The authors have declared that no competing interests exist.

Direct contact with swine infected by Aujeszky's disease virus (ADV) represents a potential risk of transmission to carnivore species, in which the infection is normally fatal. We assessed exposure to ADV in hunting dogs in an area where the virus is highly endemic in wild boar populations. Two out of 466 (0.43%; 95% CI 0.00-1.02%) hunting dogs analyzed were positive by gE-bELISA, gB-bELISA and the virus neutralization test. The seroprevalence levels detected, as well as the absence of reports of clinical cases in the hunting dog groups tested, indicate limited contact of this species with ADV in the study area. Further studies are warranted to assess the pathogenicity of Aujeszky's disease virus strains infecting wild boar.

Although wild boar can act as a persistent Aujeszky's disease (AD) reservoir, limited data are available on long-term epidemiology in free-ranging wild boar living in areas where industrial swine herds are limited. Hence, this study provides crucial information, which fills this knowledge gap, on the natural dynamics of AD infection. From 3260 sera sampled during eight hunting seasons, 162 (4.97%) were tested positive. Factors, including the animal's age class, and the sampling year, had significant effects on the probability of the wild boar being seropositive, while wild boar mean abundance per area, yearly abundance and the total number of pig farms, as well as interactions among age, year and sex, were not significant. In particular, a positive trend of seroprevalence was observed over the years, with values ranging from 2.1 to 10.8%. This long-term surveillance showed an increase in seroprevalence with a higher probability of being seropositive in older individuals and the independence of wild boar seropositivity from the likelihood of contact with pigs in the area.

Despite North Carolina's long history with feral swine, populations were low or absent in eastern counties until the 1990s. Feral swine populations have since grown in these counties which also contain a high density of commercial production swine (CPS) facilities. Sixteen of the highest swine producing U.S. counties also populated with feral swine are in North Carolina. Disconcertingly, since 2009, positive tests for exposure to swine brucellosis or pseudorabies virus have been found for feral swine. We surveyed 120 CSP facilities across four eastern counties to document the level and perception of feral swine activity around CSP facilities and to identify disease transmission potential to commercial stock. Nearly all facility operators (97%) recognized feral swine were in their counties. Far fewer said they had feral swine activity nearby (18%). Our inspections found higher presence than perceived with feral swine sign at 19% of facilities where operators said they had never observed feral swine or their sign. Nearly 90% expressed concern about feral to domestic disease transmission, yet only two facilities had grain bins or feeders fenced against wildlife access. Due to increasing feral swine populations, recent evidence of disease in feral populations, the importance of swine production to North Carolina's economy and the national pork industry, and potential for feral-domestic contact, we believe feral swine pose an increasing disease transmission threat warranting a stringent look at biosecurity and feral swine management at North Carolina CPS facilities.

In 2007, two new populations of feral swine were discovered in Nance and Valley counties, Nebraska, USA. Necropsies and serologic testing was done on two individuals from the Nance County herd. Results indicated that a lactating sow had positive antibodies for pseudorabies virus (PRV). Investigations conducted by Nebraska Game and Parks Commission Law Enforcement division confirmed that the infected individual was transported illegally to Nebraska, USA, from Texas, USA. All domestic swine herds located within an 8 km radius of the infected individual tested negative for antibodies to PRV. Our results provide a clear example of how diseases can spread because of anthropogenic activities and highlight the need for disease surveillance and monitoring in the import of invasive species.

Feral swine (Sus scrofa) are present in 38 of the 50 United States, and their populations continue to expand. Domestic swine are widely regarded as vulnerable to diseases harbored by feral swine. Our objectives were to determine antibody prevalence for selected pathogens in Texas feral swine populations and identify contact events between feral and domestic swine. Overall prevalence of antibodies against brucellosis and pseudorabies virus was 11% and 30%, respectively. Antibodies to porcine reproductive and respiratory disease virus were detected in 3% of feral swine from southern Texas. All samples tested negative for antibodies to classical swine fever virus. To determine the frequency of contact events between feral swine and domestic swine in neighboring facilities, we analyzed movement data from 37 adult feral swine that were trapped < or =10 km from domestic swine facilities and equipped with geographic positioning system collars. Seven of the 37 feral swine had contact (relocated within 100 m) with domestic swine. We found that contact between feral swine and domestic swine occurred predominantly at night. Additionally, we analyzed 60 consecutive days of experimental track plots around pens that contained domestic swine and empty control pens, and found greater visitation by feral swine to the domestic swine pens. Our data demonstrate that feral swine have direct contact with domestic swine, which presents opportunity for disease transmission.

Blood samples were collected from wild boar (Sus scrofa) shot during the hunting season from 1999 to 2005 in the Czech Republic. Sera were tested by enzyme-linked immunosorbent assay for the presence of antibodies against classical swine fever virus (CSFV), swine vesicular disease virus (SVDV), Aujeszky's disease virus (ADV), and bovine viral diarrhea virus (BVDV). Indirect fluorescence antibody test was used for detection of antibodies against porcine circovirus type 2 (PCV-2) and transmissible gastroenteritis virus (TGEV). Antibodies against ADV, BVDV, PCV-2, and TGEV were detected in 30% (101 of 338), 1% (2 of 352), 43% (57 of 134), and 1% (1 of 134) of wild boars, respectively. Sera of 6,471 and 362 tested wild boars were negative for the presence of antibodies against CSFV and SVDV, respectively. This is the first survey of TGEV antibodies in wild boars and the first serologic survey of viral diseases in wild boars in the Czech Republic. Wild boars in the Czech Republic may act as a potential reservoir of ADV and thus have a role in the epidemiology of this disease.

Tissue and blood samples were collected from 152 wild boars (Sus scrofa) from the Maremma area (Grosseto district, Central Italy) between November 2002 and January 2003. The presence of pseudorabies virus (PRV) antibodies, antigen, and DNA were confirmed by an enzyme-linked immunosorbent assay (ELISA), immunohistochemistry, and polymerase chain reaction (PCR), respectively. Of 152 animals, 62 (41%) were positive for viral antigen in tonsillar tissue. Of the 80 serum samples that were suitable for testing, 41 (51%) were positive for PRV antibodies. Positive immunohistochemistry results were confirmed by PCR. A significantly higher prevalence of PRV antigen and seroprevalence was detected in older animals. No differences were detected between males and females or for animals coming from different areas sampled. Results confirm that PRV is endemic in this wild boar population with a high prevalence of infection. The results of immunohistochemistry investigations demonstrated that a large number of wild boars harbor PRV in tonsillar tissues and should be considered as an important reservoir of PRV.

Serum samples from 427 hunter-killed wild boar (Sus scrofa) from Slovenia were tested for antibodies to Aujeszky's disease virus (ADV). Samples were collected throughout Slovenia and corresponded to 6.2% of the total harvest. Antibodies against ADV were detected in 111 sera (26%) using a commercial enzyme-linked immunosorbent assay (ELISA). Antibody prevalence increased significantly with age. This report describes the first evidence of ADV infection in wild boar populations in Slovenia.

Serologic surveys for evidence of exposure to pseudorabies virus (PRV) in feral swine were conducted from November 2001 to April 2002 at 10 sites in the southeastern United States, where evidence of previous PRV exposure had been documented during 1979-89. Sera were tested in the field on the day of collection by latex agglutination. Maximum sample size per site was to be 30 animals, but sampling was discontinued before reaching this number when positive results were obtained. Positive results were obtained at all of the study sites, demonstrating long-term persistence of PRV in feral swine populations. Overall, 38 of 100 (38%) animals were positive for antibodies. Consistent results from latex agglutination tests conducted in the field and laboratory demonstrated that this test was useful as a rapid and reliable diagnostic tool when used in the field.

Free-ranging feral swine (Sus scrofa) are known to be present in at least 32 states of the USA and are continuously expanding their range. Infection with pseudorabies virus (PRV) occurs in feral swine and the primary route of transmission in free-living conditions seems to be venereal. Between 1995 and 1999, naturally infected feral swine and experimentally infected hybrid progeny of feral and domestic swine, were kept in isolation and evaluated for occurrence of latent PRV indigenous to feral swine in sacral and trigeminal ganglia and tonsil. Sacral ganglia were shown, by polymerase chain reaction (PCR) amplification of the thymidine kinase (TK) gene of PRV, to be the most frequent sites of latency of PRV. Nine (56%) of 16 sacral ganglia, seven (44%) of 16 trigeminal ganglia, and five (39%) of 13 tonsils from naturally infected feral swine were positive for PCR amplification of TK sequences of PRV. These tissues were negative for PRV when viral isolation was attempted in Vero cells. DNA sequencing of cloned TK fragments from the sacral ganglia of two feral swine, showed only one nucleotide difference between the two fragments and extensive sequence homology to fragment sequences from various domestic swine PRV strains from China, Northern Ireland, and the USA. The hybrid feral domestic swine, experimentally inoculated with an indigenous feral swine PRV isolate by either the genital or respiratory route, acquired the infection but showed no clinical signs of pseudorabies. Virus inoculated into either the genital or respiratory tract could, at times, be isolated from both these sites. The most common latency sites were the sacral ganglia, regardless of the route and dose of infection in these experimentally infected hybrids. Nine of 10 sacral ganglia, six of 10 trigeminal ganglia, and three of 10 tonsils were positive for PCR amplification of TK sequences. No virus was isolated from these tissues in Vero cells. The demonstration of the sacral ganglia as the most common sites of latency of pseudorabies viruses indigenous to feral swine, supports the hypothesis that these viruses are primarily transmitted venereally, and not by the respiratory route as is common in domestic swine, in which the trigeminal ganglia are the predominant sites of virus latency.

Two hundred twenty seven adult (> 8 mo) feral swine (Sus scrofa) trapped from April through July 1999 at three locations on a coastal South Carolina (USA) peninsula with restricted ingress and egress were tested for Brucella suis and pseudorabies virus (PRV) antibodies. Approximately 44% of the animals tested positive for B. suis antibodies and 61% tested positive for antibodies to PRV. Previous surveys (1976 and 1992) of feral swine at the same location with similar methods indicated lower seroprevalences (28% and 18% for B. suis and 0% and 19% for PRV). We also found 39% of feral swine seropositive (n = 179) for Trichinella spiralis and 49% seropositive (n = 181) for Toxoplasma gondii. Results of repeated sampling demonstrated that seroprevalence to pathogens can increase with time in an isolated, unhunted population of feral swine suggesting an increased risk to local domestic livestock and potentially to human health.

The determination of serologic titers to infectious organisms is a valuable tool for quantitating exposure to disease organisms. Raccoons (Procyon lotor) were live-trapped from September 1989 to October 1993 and samples collected from two distinct locations in west-central Illinois (USA); a state recreational facility (Park) and privately owned farming property (Farm). Sera were submitted for testing Leptospira interrogans (serovars bratislava, canicola, grippotyphosa, hardjo, icterohemmorhagiae, and pomona), canine distemper virus (CDV), pseudorabies virus (PV), and Toxoplasma gondii. Two-hundred and twenty-two (48%) of 459 raccoons were seropositive for L. interrogans. Eighty-five (23%) out of 368 raccoons were seropositive for canine distemper virus. Eighty-two (17%) of 479 raccoons raccoons were seropositive for pseudorabies virus. One hundred and eight-four (49%) of 379 raccoons were seropositive for T. gondii. A significant difference (P < 0.05) in seroprevalence for L. interrogans between the park (43%) and farm (52%) areas was found. A correlation between increasing age and seroprevalence was found for L. interrogans, CDV, PV, and T. gondii. Furthermore, there was a significant difference in seroprevalence for T. gondii during the spring trapping seasons (73%), when compared with the fall (33%). This type of information on exposure to infectious agents is important for developing control programs to manage raccoon-human and raccoon-domestic animals interactions.

Sera collected from European wild boar (Sus scrofa) shot in Eastern Germany between January 1991 and December 1994 were tested for antibodies to pseudorabies virus (PRV). Of 3,143 sera tested, 281 (8.9%) and 13 (0.4%) were positive and suspect in an enzyme-linked immunosorbent assay (ELISA), respectively. The specificity of the reactions was confirmed by detection of neutralizing antibodies in 220 sera (74.8%) and by immunoblotting. Analysis of host age and sex of the animals, temporal and spatial factors showed significantly higher seroprevalences in older animals than in younger individuals, but no differences between males and females. Pseudorabies virus infections have been endemic in this wild boar population for several years and the extreme eastern part of the study area had significantly higher seroprevalences (< or = 22%) than other areas. In the area covered by this study, pseudorabies virus was eradicated in the domestic animal populations in 1985. Thus, the infections in the wild boar population appear to be endemic and persist completely separately and without affecting the domestic pig population.

Pseudorabies (Aujeszky's disease) was diagnosed in three adult captive coyotes (Canis latrans) from southern Indiana (USA). The coyotes died in their outdoor enclosure within a 48 hr period. Histopathology revealed multifocal, nonsuppurative meningioencephalitis and eosinophilic intranuclear inclusion bodies within neurons. Samples of brain were positive for pseudorabies virus by fluorescent antibody testing and virus isolation. Source of infection was the probable consumption of pseudorabies virus-infected pig carcasses.

An epizootic of Aujeszky's disease (pseudorabies) in four captive European brown bears (Ursus arctos) in November 1994, in the Val di Non, Trentino Region, Italy, was linked to consumption of raw pork. Affected animals had severe pruritus resulting in self-mutilation, and all four died within 24 hr after onset of clinical signs. Aujeszky's disease virus was isolated on first passage from the brain and was characterized by means of restriction endonuclease analysis. Based on these data, we believe that bears are extremely susceptible to the disease, and that wildlife managers should consider pseudorabies as a potential risk for the captive and wild bear populations.

A Gaussian diffusion model was applied to an epizootic of pseudorabies in ten swine herds located in Decatur County, Indiana, USA to test the hypothesis that the virus can be spread via aerosol. The epizootic occurred during January to March, 1988, spreading through ten farms across an area of about 150 km2. The model included a receptor component that provided an estimate of viruses received by the pig within an enclosed barn. Results show that the diffusion model can explain the spread of the virus during the epizootic for all nine farms to which the virus spread.

Blood samples were collected from 108 wild hogs (Sus scrofa) from the Great Smoky Mountains National Park (GSMNP), USA, February to July 1990. We found no antibodies for swine brucellosis, pseudorabies, bovine virus diarrhea virus or porcine rotavirus infection. Antibody titers to porcine parvovirus were found in 15 (14%) samples and antibody to one or more leptospiral serovars was found in 48 (44%) samples. Thirty-nine (89%) of the 44 positive samples reacted to all five leptospiral serovars tested.

Serum samples collected from feral swine (Sus scrofa) throughout Florida (USA) from 1980 to 1989 were tested for antibodies to pseudorabies virus (PRV) by the serum neutralization test, the latex agglutination test, or by the enzyme-linked immunosorbent assay. Seropositive swine were detected at 11 of 13 sites with a composite seroprevalence of 34.8% (579 of 1,662 samples; range = 5.9% to 58.2%) for sites with seropositive swine. Data on age and sex of the swine were available from three sites. Seroprevalence in males and females did not differ significantly (P = 0.62 for the combined data). Seroprevalence in adult (> or = 8 mo) and juvenile swine (< 8 mo) was significantly different at all sites (P < 0.05 for the combined data). From these data, PRV infections appear to occur widely in populations of Florida feral swine and may seriously undermine efforts to eradicate this virus from the domestic swine population of the USA.

Six hundred sixty-one feral swine (Sus scrofa) from Ossabaw Island, Georgia (USA) were captured, bled, and their sera tested for pseudorabies virus (PRV) antibody during a 6 yr period. Prevalence of seroconversion in females was somewhat higher than in males (10% versus 7%), but the difference was not statistically significant. Adults had a significantly higher prevalence than juveniles (29% versus 1%). An important finding in this study was that seroconversion occurred primarily in the adult feral swine.