Aujeszky's disease in hunting dogs after the ingestion of wild boar raw meat in Sicily (Italy): clinical, diagnostic and phylogenetic features.

BACKGROUND: Aujeszky's disease is caused by Suid Herpes Virus-1 and species belonging to the genus Sus scrofa are the main reservoir hosts. This virus, however, is capable of infecting and causing severe disease, with an almost constant fatal outcome in other species, both domestic and wild (carnivores, monogastric herbivores and ruminants). Moreover, the possibility of transmission to humans has been demonstrated. This study reports and describes the clinical, diagnostic, pathological and phylogenetic aspects of two cases of Aujeszky's disease in two hunting dogs following the ingestion of infected wild boar raw meat. These cases are contextualized in the province of Messina (Sicily), where a high prevalence of Aujeszky's disease has been recorded (average of 12,20% in the period 2010-2019) in farmed pig, and with evidence of spread to other species. A severe outbreak in cattle has recently been reported in these areas. Nevertheless, cases of Aujeszky's disease in dogs are rarely reported and this study represents the first well-documented report in this species in Sicily. CASE PRESENTATION: After a wild boar hunt, two dogs showed neurological symptoms and intense itching unresponsive to therapy. Diagnosis of Aujeszky's disease was made based on clinical suspicion, anamnestic information and confirmed by the isolation of the virus from the brain of both dogs. In addition, molecular typing, sequencing and phylogenetic analysis of the Real-Time PCR products were performed. The sequences studied were placed in the Italian Clade 1 along with the sequences obtained from wild boars and hunting dogs from Italy and France. CONCLUSIONS: The finding of this disease in non-natural hosts in Sicilian multi-host epidemiological contexts suggests that the risk of inter-species transmission is concrete and that attention should be paid to developing disease control programs in these territories. The data obtained from genome sequencing of the two SuHV-1 isolates contribute to the enrichment of the GenBank with unknown sequences and the phylogenetic analysis implementation.

Evidence for interspecies transmission route of pseudorabies virus via virally contaminated fomites.

Pseudorabies virus (PRV) is a zoonotic agent with a wide host range, causing significant economic losses in animal husbandry and potential public health risk globally. The causative agent has recently gained attention due to the inter-species transmission among different species of animals, even human beings. Although PRV's prevalence is found in many species of animals, regardless of whether the strain involved is a vaccine, classical or variant, few lines of evidence for the viral transmission route are available. Here, we reported that viral contamination is associated with the inter-species transmission of PRV. We found that PRV contamination was widely distributed in the environment of pig farms, that viral distribution in the environment is associated with the implementation of biosecurity measures, and that PRV could transmit from pigs to dogs through virally contaminated fomites. Collectively, our findings provide a basis for understanding the ecology and transmission route of PRV and underscore the importance of implementing biosecurity measures to control this disease.

Characteristics of human encephalitis caused by pseudorabies virus: A case series study.

BACKGROUND: Pseudorabies virus (PRV) has been thought to cause diseases only in animals. However, recent studies have shown that PRV can also cause illnesses in humans. METHODS: This was a case series study. The cases of five patients with clinical symptoms of acute encephalitis, which were confirmed to be caused by PRV infections, were reviewed. CASE PRESENTATION: The five patients all had jobs involving the handling of pigs. They had acute onset and rapid progression of clinical presentations, which were consistent with central nervous system infections. Four of them had respiratory failure, which required ventilator support. Brain magnetic resonance imaging showed abnormal signals in the bilateral temporal lobes and insular cortex in all five patients, bilateral frontal lobes in one patient, and caudate nucleus in one patient. Cerebrospinal fluid analysis results were consistent with a viral infection. Next-generation sequencing of the cerebrospinal fluid confirmed the presence of PRV. All patients received human immunoglobulin, glucocorticoids, antiviral agents, and symptomatic supportive treatments. All patients survived until discharge, but suffered from various sequelae. Pneumonia was the most common complication during the disease course. CONCLUSIONS: PRV encephalitis should be included in the differential diagnosis of patients with a clinical presentation of central nervous system infection, especially for those who have had recent contact with pigs.

Dynamic cerebrospinal fluid analyses of severe pseudorabies encephalitis.

We reported a severe human pseudorabies encephalitis case and described a dynamic clinical manifestation with cerebrospinal fluid analyses and cytological and serological evaluation, which may elucidate the mechanism of PRV infection and facilitate clinical diagnosis and treatment in human.

Survey of Aujeszky's Disease Virus in Hunting Dogs from Spain.

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.

Commercial vaccine against pseudorabies virus: A hidden health risk for dogs.

Pseudorabies virus (PRV) is considered as an infectious agent with a wide of host range, causing considerable economic losses in animal husbandry. Although the commercial vaccine against PRV plays an critical role in control of this disease in swine industry, the potential risk of commercial vaccines against PRV for other host is unclear. Here, we report that the commercial vaccine against PRV is a hidden health risk for dogs. We found that different attenuated PRV strains in commercial vaccines possess different tissue tropism, and that the attenuated PRV strains are lethal to dogs, and that the attenuated PRV strain possesses the ability to spread horizontally among the dogs. Collectively, our findings provide clues that the commercial vaccine against PRV is a hidden risk for dogs, even for the owner of pet dogs to take seriously.

Isolation and molecular characterization of a variant of Chinese gC-genotype II pseudorabies virus from a hunting dog infected by biting a wild boar in Japan and its pathogenicity in a mouse model.

We isolated a variant of Chinese pseudorabies virus from a hunting dog with symptoms similar to Aujeszky's disease and designated the isolate MY-1 strain. The dog developed symptoms 6 days after hunting and biting a wild boar and died the day after onset. The Bam HI restriction profile of MY-1 DNA was different from those of the Japanese reference strain Yamagata-S81 and two vaccine strains, Bartha and Begonia, and resembled Bam HI-RFLP (restriction fragment length polymorphism) type IV. Complete nucleotide sequences were determined, and phylogenetic analyses revealed that MY-1 belonged to the same cluster of old Chinese strains and variant strains isolated recently in China, but most of the open reading frames of MY-1 were located on a different branch from those of these Chinese strains. Based on a gC phylogenetic analysis, MY-1 belonged to gC-genotype II composed of those Chinese strains. In mice, the 50% lethal dose (LD50) of MY-1 (103.0 TCID50) was almost the same as those of Yamagata-S81 and Bartha. The LD50 value of Begonia was 10≥4.5 TCID50. The mean survival periods of mice after infection with 104 TCID50 of MY-1, Yamagata-S81 and Bartha were 3.9 days, 2.3 days, and 8.0 days, respectively. The results suggested that the variant of Chinese PRV with slightly weaker pathogenicity than that of wild virulent viruses might be maintained in wild boars in Japan. Furthermore, we would like to propose that old Chinese strains, recent Chinese variant strains, and MY-1 should be grouped as an Asian type PRV.

Interspecies Transmission, Genetic Diversity, and Evolutionary Dynamics of Pseudorabies Virus.

BACKGROUND: Pseudorabies virus (PRV) causes Aujeszky's disease in pigs and can be transmitted to other mammals, including humans. In the current study, we systematically studied the interspecies transmission and evolutionary history of PRV. METHODS: We performed comprehensive analysis on the phylodynamics, selection, and structural biology to summarize the phylogenetic and adaptive evolution of PRV based on all available full-length and major glycoprotein sequences. RESULTS: PRV can be divided into 2 main clades with frequent interclade and intraclade recombination. Clade 2.2 (variant PRV) is currently the most prevalent genotype worldwide, and most commonly involved in cross-species transmission events (including humans). We also found that the population size of clade 2.2 has increased since 2011, and the effective reproduction number was >1 from 2011 to 2016, indicating that PRV may be still circulating in swine herds and is still a risk in relation with cross-species transmission in China. Of note, we identified amino acid sites in some important glycoproteins gB, gC, gD, and gE that may be associated with PRV adaptation to new hosts and immune escape to vaccines. CONCLUSIONS: Our study provides important genetic insight into the interspecies transmission and evolution of PRV within and between different hosts that warrant additional surveillance.

Pseudorabies detected in hunting dogs in Alabama and Arkansas after close contact with feral swine (Sus scrofa).

BACKGROUND: Pigs (Sus scrofa) are the natural hosts of pseudorabies virus (PRV), also known as Aujeszky's disease. Infection in mammals, with the exception of humans, typically causes extreme itching, facial swelling, and excessive salivation, followed by death in non-suid species. The risk to susceptible mammals was assumed to decrease when PRV was eliminated from U.S. commercial swine in 2004, though the virus remains endemic in feral swine. Infected feral swine pose a threat to the disease-free status of the commercial swine industry, and to other animals, including dogs, that come in direct or indirect contact with them. Since dogs are commonly used for hunting feral swine, they are at high risk of exposure. CASE PRESENTATION: The following report describes the progression of pseudorabies infection in dogs in two states after exposure to feral swine. The first case occurred in a dog in Alabama after participation in a competitive wild hog rodeo. The second case occurred in multiple dogs in Arkansas after hunting feral swine, and subsequent consumption of the offal. The antibody prevalence of feral swine in the two states where the dogs were exposed is also examined. CONCLUSIONS: Dogs that are used for hunting feral swine are at high risk of exposure to pseudorabies because the disease is considered endemic in feral swine in the U.S.

Detección serológica de Brucella suis, virus de influenza y virus de la enfermedad de Aujeszky en criaderos porcinos familiares de menos de 100 madres en Argentina / Serological detection of Brucella suis, influenza virus and Aujeszky's disease virus in backyard and small swine holders in Argentina

Los criaderos porcinos de menos de 100 madres representan más del 99% de los de todo el país; sin embargo, existen escasos reportes sobre su situación sanitaria y productiva. Se recabó información productiva y se tomaron muestras para detectar anticuerpos contra Brucella suis (Bs), virus de la enfermedad de Aujeszky (VA) y virus de influenza (VI) en 68 establecimientos de menos de 100 madres ubicados en la región norte, centro y sur del país. El 80% de los establecimientos fueron positivos al VI H1 pandémico 2009, el 11% al H3 clúster 2, mientras que el 11,7% presentó anticuerpos contra el VA y el 6% contra Bs. Ninguno de los productores conocía los factores de riesgo para la transmisión de enfermedades del cerdo al humano. El 47% compra sus reproductores a pares o en ferias. En lo que respecta a normas de bioseguridad, solo el 16% de los establecimientos tenía cerco perimetral y el 37% de las granjas contaba con asesoramiento veterinario. Los resultados de este estudio demuestran que la caracterización productiva y el relevamiento sanitario son de suma importancia para mejorar la productividad y reducir el riesgo de transmisión de enfermedades. El conocimiento de la situación sanitaria y de los factores de riesgo es necesario para conseguir un mejor control y la erradicación de enfermedades en sistemas de baja tecnificación. Se deberían llevar a cabo estudios más representativos a nivel país para detectar los agentes circulantes y, sobre la base de esta información, implementar medidas de prevención y control.

Farmers raising less than 100 sows represent more than 99% of swine producers in Argentina, although little is known about their sanitary status and productive characteristics in the country. Sanitary and productive information was obtained. Furthermore, samples for serological studies were taken to detect antibodies against Brucella suis (Bs), Aujeszky's disease virus (AV) and influenza virus (IV) in 68 backyard and small producers with less than 100 sows located in the north, central and south regions of Argentina. Antibodies against H1 pandemic were detected in 80% of the farms while 11%, 11.7% and 6.0% of the producers were positive to influenza H3 cluster 2, AV and Bs, respectively. None of the producers was aware of the risk factors concerning the transmission of diseases from pigs to humans. A percentage of 47% of them buy pigs for breeding from other farmers and markets. With regard to biosecurity measures, only 16% of the farms had perimeter fences. The results of this study demonstrate that productive characterization and disease surveys are important to improve productivity and to reduce the risk of disease transmission among animals and humans. The study of sanitary status and risk factors is necessary for better control and eradication of diseases in backyard or small producers. More representative studies at country level should be carried out to detect the pathogensthat circulate and, with this knowledge, to implement prevention and control measures.

[Serological detection of Brucella suis, influenza virus and Aujeszky's disease virus in backyard and small swine holders in Argentina]. / Detección serológica de Brucella suis, virus de influenza y virus de la enfermedad de Aujeszky en criaderos porcinos familiares de menos de 100 madres en Argentina.

Farmers raising less than 100 sows represent more than 99% of swine producers in Argentina, although little is known about their sanitary status and productive characteristics in the country. Sanitary and productive information was obtained. Furthermore, samples for serological studies were taken to detect antibodies against Brucella suis (Bs), Aujeszky's disease virus (AV) and influenza virus (IV) in 68 backyard and small producers with less than 100 sows located in the north, central and south regions of Argentina. Antibodies against H1 pandemic were detected in 80% of the farms while 11%, 11.7% and 6.0% of the producers were positive to influenza H3 cluster 2, AV and Bs, respectively. None of the producers was aware of the risk factors concerning the transmission of diseases from pigs to humans. A percentage of 47% of them buy pigs for breeding from other farmers and markets. With regard to biosecurity measures, only 16% of the farms had perimeter fences. The results of this study demonstrate that productive characterization and disease surveys are important to improve productivity and to reduce the risk of disease transmission among animals and humans. The study of sanitary status and risk factors is necessary for better control and eradication of diseases in backyard or small producers. More representative studies at country level should be carried out to detect the pathogensthat circulate and, with this knowledge, to implement prevention and control measures.

Outbreak of severe pseudorabies virus infection in pig-offal-fed farmed mink in Liaoning Province, China.

An outbreak of severe pseudorabies virus (PRV) infection in farmed mink occurred in northern China in late 2014, causing significant economic losses in the local fur industry. Here, we report the first case of a PRV outbreak in mink in northeastern China, caused by feeding farmed mink with raw pork or organs contaminated by PRV. Mink infected with virulent PRV exhibited diarrhea, neurologic signs, and higher mortality, which can be misdiagnosed as highly pathogenic mink enteritis virus (MEV), canine distemper virus (CDV), and food poisoning. However, these were excluded as causative agents by PCR or bacteria isolation. The duration of disease was 3-7 days, and the mortality rate was 80-90%. PRV was characterized using indirect immunofluorescence assays (IFA) and electron microscopy (EM). Phylogenetic analysis based on full-length genome sequences and those of individual genes of this novel virus strain showed that it clustered in an independent branch with several other PRV isolates from China.

Age- and strain-dependent differences in the outcome of experimental infections of domestic pigs with wild boar pseudorabies virus isolates.

Although pseudorabies virus (PRV) has been eradicated in domestic swine in many countries, its presence in wild boars remains a threat for a reintroduction into the currently unprotected swine population. To assess the possible impact of such a reintroduction in a naive herd, an in vivo infection study using two genetically characterized wild boar PRV isolates (BEL24043 and BEL20075) representative for wild boar strains circulating in south-western and central Europe and the virulent NIA3 reference strain was performed in 2- and 15-week-old domestic pigs. Our study revealed an attenuated nature of both wild boar strains in 15-week-old pigs. In contrast, it showed the capacity of strain BEL24043 to induce severe clinical symptoms and mortality in young piglets, thereby confirming that the known age dependency of disease outcome after PRV infection also holds for wild boar isolates. Despite the absence of clinical disease in 15-week-old sows, both wild boar PRV strains were able to induce seroconversion, but to a different extent. Importantly, differences in infection and transmission capacity of both strains were observed in 15-week-old sows. Strain BEL24043 induced a more prolonged and disseminated infection than strain BEL20075 and was able to spread efficiently to contact animals, indicative of its capacity to induce a sustained infection. In conclusion, it was shown that a reintroduction of a wild boar isolate into the domestic swine population could have serious economic consequences due to the induction of clinical symptoms in piglets and by jeopardizing the PRV-negative status.

Molecular characterization of Belgian pseudorabies virus isolates from domestic swine and wild boar.

Aujeszky's disease is an economically important disease in domestic swine caused by suid herpesvirus 1, also called pseudorabies virus (PRV). In several European countries, including Belgium, the virus has successfully been eradicated from the domestic swine population. The presence of PRV in the wild boar population however poses a risk for possible reintroduction of the virus into the domestic pig population. It is therefore important to assess the genetic relatedness between circulating strains and possible epidemiological links. In this study, nine historical Belgian domestic swine isolates that circulated before 1990 and five recent wild boar isolates obtained since 2006 from Belgium and the Grand Duchy of Luxembourg were genetically characterized by restriction fragment length polymorphism (RFLP) analysis and phylogenetic analysis. While all wild boar isolates were characterized as type I RFLP genotypes, the RFLP patterns of the domestic swine isolates suggest that a shift from genotype I to genotype II might have occurred in the 1980s in the domestic population. By phylogenetic analysis, Belgian wild boar isolates belonging to both clade A and B were observed, while all domestic swine isolates clustered within clade A. The joint phylogenetic analysis of both wild boar and domestic swine strains showed that some isolates with identical sequences were present within both populations, raising the question whether these strains represent an increased risk for reintroduction of the virus into the domestic population.

Serological and molecular investigation of the prevalence of Aujeszky's disease in feral swine (Sus scrofa) in the subregions of the Pantanal wetland, Brazil.

The feral swine (FS) originated from the domestic pig and is present throughout the Brazilian wetland plain (the Pantanal). Aujeszky's disease (AD) was first serologically confirmed in the state of Mato Grosso do Sul (MS) in 2001; however, there was no viral confirmation. The aim of this study was to investigate antibodies against-SuHV-1 in the sera of feral swine in the studied areas, detect SuHV-1 through PCR and classify the viral genome. Among the 218 animals sampled, 186 were analyzed by ELISA, resulting in 88 (47.3%) reactive samples. In the serum neutralization test (SN), 57/179 (31.8%) samples presented antibodies against the AD virus (SuHV-1). By nested PCR, 104 DNA samples were extracted for analysis and confirmed with amplification of a fragment of glycoprotein B (gB) in five samples. The SuHV-1 was detected in 12 samples by using primers for glycoprotein E (gE) and viral genome was classified as Type I by ul44 partial sequencing. The amplification of SuHV-1 glycoprotein fragments in the fetuses of seropositive sows indicate that the vertical transmission contribute to maintain SuHV-1 in a free-living feral swine population. The origin of AD in the feral swine populations of the Pantanal is unknown, however, the determination of viral latency, the vertical transmission of the antigen by the amplification of SuHV-1 glycoprotein fragments in the fetuses of seropositive sows and genome typing contribute to the elucidation of the epidemiology of this disease in the wetlands of MS, Brazil.

Minimization of the impact of Aujeszky's disease outbreaks in The Netherlands: a conceptual framework.

In the Netherlands, outbreaks of Aujeszky's Disease (AD) are controlled by vaccination and movement restriction zones (MRZ). Although this strategy avoids the socio-ethical concerns associated with pre-emptive slaughter, it can easily result in animal welfare problems and negative economic consequences. These arise because movement restrictions result in surpluses of live (vaccinated) piglets on farms. The aim is to provide insight into the development of these surpluses and its impact and to describe how measures that allow early transportation of pigs under certain conditions and to specific destinations (channelling) could reduce these problems. For the analysis, a deterministic simulation model was developed, which calculates surpluses of piglets at multiplier farms during AD outbreaks. This is performed on a weekly basis for two areas (with and without piglet surplus), three outbreak durations (minimum, moderate and long) and three strategies for movement restrictions (strict, transports within the MRZ allowed and transports outside the MRZ allowed). The results show that in case of complete movement restrictions, surpluses of piglets varying in age and vaccination status will quickly arise. These surpluses are larger for longer epidemics and can become as large as 180-340 thousand piglets (45-75% of weekly domestic production) for moderate and long epidemics, respectively. Implementation of channelling strategies that allow earlier transportation within the MRZ can reduce surpluses by about 50% to 100-150 thousand piglets maximum. Strategies that also allow transportation outside the MRZ can reduce surpluses even further to below 100 thousand piglets. It was concluded that channelling of live piglets during AD outbreaks results in a drastic reduction of problems with accommodating ready-for-transport piglets. Moreover, it reduces shortages during movement restrictions and peak supply immediately after removing the restrictions. Channelling could therefore be an important instrument to reduce the economic and animal welfare impacts of containment measures.

Wild boar: an increasing concern for Aujeszky's disease control in pigs?

BACKGROUND: The goal of this study was describing the temporal evolution of Aujeszky's disease virus (ADV) contact prevalence among Eurasian wild boar (Sus scrofa) populations under different management regimes and contact likelihoods with domestic pigs. Given the recent increase in wild boar abundance throughout Europe, we hypothesized that wild boar contact with ADV would remain stable in time even after significant reduction of ADV prevalence in domestic pigs. RESULTS: Sera from 1659 wild boar were collected from 2000 to 2010 within 6 areas of the Iberian Peninsula and tested for the presence of antibodies against ADV by ELISA. According to sampling date, wild boar were grouped into three time periods. ADV prevalence was compared through period both globally and by geographic area. Overall seroprevalence for the ten-year study period was 49.6 ± 2.4%. The highest seroprevalence was recorded in areas with intense wild boar management. The annual proportion of positive wild boar sampling sites remained stable through the study period, while the percentage of domestic pig AD positive counties decreased from 70% in 2003 to 1.7% in 2010. CONCLUSIONS: Results presented herein confirmed our hypothesis that ADV would remain almost stable in wild boar populations. This evidences the increasing risk wild boar pose in the final stages of ADV eradication in pigs and for wildlife conservation.

Preferential sexual transmission of pseudorabies virus in feral swine populations may not account for observed seroprevalence in the USA.

This paper compares the behavior of two competing models for the transmission of pseudorabies virus in feral swine in the USA. In first model, horizontal (non-sexual) density dependent transmission is the only transmission modality. In the second model, the only transmission modality is sexual transmission between mature males and females. The comparison of model behavior was carried out to test the hypothesis that preferential sexual transmission of PRV in feral swine can account for the seroprevalence observed in the field. The observed range of seroprevalence of PRV in mature feral swine in the USA is consistent with a preferential sexual transmission only if the feral swine mating system is a random mating system or a polygynous system in which there is a relatively large rate of acquisition of new mates. The observed range of seroprevalence of PRV in mature feral swine in the USA is not consistent with a preferential sexual transmission if there is mate guarding. This is important because the National Pseudorabies Surveillance Plan deems monitoring the risk of PRV introduction from feral swine to be a "minor objective" both in terms of the scope of the plan and with respect to the resources allocated. The rationale for this statement was derived from experimental studies, which suggested that the PRV indigenous to feral swine in the USA is preferentially sexually transmitted.

Making contact: rooting out the potential for exposure of commercial production swine facilities to feral swine in North Carolina.

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.