Pigs in southern Italy are exposed to three ruminant pathogens: an analysis of seroprevalence and risk factors analysis study.

BACKGROUND: Pigs are susceptible to several ruminant pathogens, including Coxiella burnetti, Schmallenberg virus (SBV) and bovine viral diarrhea virus (BVDV). These pathogens have already been described in the pig population, although the dynamics of the infection and the impact on pig farms are currently unclear. The aim of this work was to evaluate the presence of these infections in the pig population of the Campania region, southern Italy, and to evaluate the risk factors associated with a greater risk of exposure. RESULTS: A total of 414 serum samples belonging to 32 herds were tested for the presence of antibodies against SBV, Coxiella, and BVD using commercial multispecies ELISA kits. SBV (5.3%) was the most prevalent pathogen, followed by Coxiella (4.1%) and BVD (3%). The risk factors included in the study (age, sex, province, farming system, ruminant density and major ruminant species) had no influence on the probability of being exposed to BVD and Coxiella, except for the location, in fact more pigs seropositive to Coxiella were found in the province of Caserta. However, the univariate analysis highlighted the influence of age, location, and sex on exposure to SBV. The subsequent multivariate analysis statistically confirmed the importance of these factors. The presence of neutralizing antibodies for SBV and BVDV, or antibodies directed towards a specific phase of infection for Coxiella was further confirmed with virus-neutralization assays and phase-specific ELISAs in a large proportion of positive samples. The presence of high neutralizing antibody titers (especially for SBV) could indicate recent exposures. Twelve of the 17 positive samples tested positive for antibodies against Coxiella phase I or II antigens, indicating the presence of both acute and chronic infections (one animal tested positive for both phases antibodies). CONCLUSIONS: Our study indicates a non-negligible exposure of pigs from southern Italy to the above pathogens. Further studies are necessary to fully understand the dynamics of these infections in pigs, the impact on productivity, and the public health consequences in the case of Coxiella.

Detection and molecular analysis of Pseudorabies virus from free-ranging Italian wolves (Canis lupus italicus) in Italy - a case report.

BACKGROUND: The only natural hosts of Pseudorabies virus (PRV) are members of the family Suidae (Sus scrofa scrofa). In mammals, the infection is usually fatal and typically causes serious neurologic disease. This study describes four Aujeszky's disease cases in free-ranging Italian wolves (Canis lupus italicus). In Italy, the wolf is a strictly protected species and is in demographic expansion. CASE PRESENTATION: Three wolves (Wolf A, B, and C) were found in a regional park in Northern Italy, and one (Wolf D) was found in Central Italy. Wolf A and D were alive at the time of the finding and exhibited a fatal infection with epileptic seizures and dyspnoea, dying after a few hours. Wolf B presented scratching lesions under the chin and a detachment of the right earlobe, whilst Wolf C was partially eaten. The wolves showed hepatic congestion, diffuse enteritis, moderate pericardial effusion, severe bilateral pneumonia, and diffuse hyperaemia in the brain. The diagnostic examinations included virological analyses and detection of toxic molecules able to cause serious neurological signs. All four wolves tested positive for pseudorabies virus (PrV). The analysed sequences were placed in Italian clade 1, which is divided into two subclades, "a" and "b". The sequences of Wolf A, B, and C were closely related to other Italian sequences in the subclade b, originally obtained from wild boars and hunting dogs. The sequence from Wolf D was located within the same clade and was closely related to the French hunting dog sequences belonging to group 4. CONCLUSION: Results showed the presence of PrV strains currently circulating in wild boars and free-ranging Italian wolves. The genetic characterisation of the PrV UL44 sequences from the four wolves confirmed the close relationship with the sequences from wild boars and hunting dogs. This fact supports a possible epidemiological link with the high PrV presence in wild boars and the possibility of infection in wolves through consumption of infected wild boar carcasses or indirect transmission. To the best of our knowledge, this study is the first detection of Pseudorabies virus in free-ranging Italian wolves in northern and central Italy.

Molecular epidemiological and genetic characterization of pseudorabies virus in Guangxi, China.

Pseudorabies virus (PRV) is an important pathogen that can cause harm to the pig population. Since 2011, there have been a number of large-scale outbreaks of pseudorabies on Chinese farms where animals had been vaccinated with the Bartha-K61 vaccine. In order to understand the epidemiological trend and genetic variations of PRV in Guangxi province, China, 819 tissue samples were collected from swine farms where PRV infection was suspected from 2013 to 2019, and these were tested for infectious wild strains of PRV. The results showed a positive rate of PRV in Guangxi province of 28.21% (231/819). Thirty-six wild-type PRV strains were successfully isolated from PRV-positive tissue samples, and a genetic evolutionary analysis was performed based on the gB, gC, gD, gE, and TK genes. Thirty of the PRV strains were found to be closely related to the Chinese variant strains HeN1-China-2012 and HLJ8-China-2013. In addition, five PRV strains were genetically related to Chinese classical strains, and one isolate was a recombinant of the PRV variant and the vaccine strain Bartha-K61. Amino acid sequence analysis showed that all 36 PRV strains had characteristic variant sites in the amino acid sequences of the gB, gC, gD, and gE proteins. Pathogenicity analysis showed that, compared to classical PRV strains, the PRV variant strains were more pathogenic in mice and had a lower LD50. Taken together, our results show that wild-type PRV infections are common on pig farms in Guangxi province of China and that the dominant prevalent strains were those of the PRV variants. The PRV variant strains also had increased pathogenicity in mice. Our data will provide a useful reference for understanding the prevalence and genetic evolution of PRV in China.

Suid alphaherpesvirus 1 of wild boar origin as a recent source of Aujeszky's disease in carnivores in Germany.

BACKGROUND: The high susceptibility of carnivores to Suid Alphaherpesvirus 1 [SuAHV1, synonymous pseudorabies virus (PrV)], renders them inadvertent sentinels for the possible occurrence of Aujeszky's disease (AD) in domestic and wild swine populations. The aim of this study was to epidemiologically analyse the occurrence of PrV infections in domestic and wild animals in Germany during the last three decades and to genetically characterise the causative PrV isolates. METHODS: PrV in dogs was detected using standard virological techniques including conventional and real time PCR, virus isolation or by immunohistochemistry. Available PrV isolates were characterized by partial sequencing of the open gC reading frame and the genetic traits were compared with those of archived PrV isolates from carnivores and domestic pigs from Germany before the elimination of AD in the domestic pig population. RESULTS: During 1995 and 2022, a total of 38 cases of AD in carnivores, e.g. dogs and red foxes, were laboratory confirmed. Sequencing and subsequent phylogenetic analysis of PrV isolates established a strong connection between AD cases in carnivores and the occurrence of PrV infections in European wild boars in the end phase of and after elimination of AD from the domestic pig population. While PrV infections occur at low numbers but regularly in hunting dogs, interestingly, PrV was not observed in grey wolves in Germany. In none of 682 dead-found grey wolves and wolf-dog hybrids tested from Germany during 2006-2022 could PrV infection be detected by molecular means. CONCLUSIONS: Although PrV has been eliminated from domestic pigs, spillover infections in domestic and wild carnivores should always be expected given the endemic presence of PrV in wild pig populations. Since detection of PrV DNA and virus in carnivores is sporadic even in areas with high seroprevalence of PrV in wild pigs, it may not reflect the full diversity of PrV.

Epidemiological Investigation, Risk Factors, Spatial-Temporal Cluster, and Epidemic Trend Analysis of Pseudorabies Virus Seroprevalence in China (2017 to 2021).

Pseudorabies virus (PRV) is a double-stranded linear DNA virus capable of infecting various animals, including humans. We collected blood samples from 14 provinces in China between December 2017 and May 2021 to estimate PRV seroprevalence. The PRV gE antibody was detected using the enzyme-linked immunosorbent assay (ELISA). Logistic regression analysis identified potential risk factors associated with PRV gE serological status at the farm level. Spatial-temporal clusters of high PRV gE seroprevalence were explored using SaTScan 9.6 software. Time-series data of PRV gE seroprevalence were modeled using the autoregressive moving average (ARMA) method. A Monte Carlo sampling simulation based on the established model was performed to analyze epidemic trends of PRV gE seroprevalence using @RISK software (version 7.0). A total of 40,024 samples were collected from 545 pig farms across China. The PRV gE antibody positivity rates were 25.04% (95% confidence interval [CI], 24.61% to 25.46%) at the animal level and 55.96% (95% CI, 51.68% to 60.18%) at the pig farm level. Variables such as farm geographical division, farm topography, African swine fever (ASF) outbreak, and porcine reproductive and respiratory syndrome virus (PRRSV) control in pig farms were identified as risk factors for farm-level PRV infection. Five significant high-PRV gE seroprevalence clusters were detected in China for the first time, with a time range of 1 December 2017 to 31 July 2019. The monthly average change value of PRV gE seroprevalence was -0.826%. The probability of a monthly PRV gE seroprevalence decrease was 0.868, while an increase was 0.132. IMPORTANCE PRV is a critical pathogen threatening the global swine industry. Our research fills knowledge gaps regarding PRV prevalence, infection risk factors, spatial-temporal clustering of high PRV gE seroprevalence, and the epidemic trend of PRV gE seroprevalence in China in recent years. These findings are valuable for the clinical prevention and control of PRV infection and suggest that PRV infection is likely to be successfully controlled in China.

Characterization of new strains of Pseudorabies virus in Argentina: Detection of interspecies transmission.

Background: Aujeszky's disease is mainly a swine disease, still endemic worldwide. It can infect other mammalians, including human beings, and it is usually fatal with nervous symptoms. Ever since the disease was detected in 1988 in Argentina, many outbreaks have been reported involving both feral swine and dogs. Aim: At present, in Argentina, Pseudorabies virus (PRV) cases are sporadically reported; however, clinical cases are informed. This study aims to obtain information about the seroprevalence of PRV in wild boars and to isolate and characterize PRV from clinical samples. Methods: From 2018 to 2019, 78 wild boars' serum samples from Bahía de Samborombón natural reserve were analyzed for antibodies to PRV using a virus neutralization test. Clinical samples from 17 pigs, 2 wild boars, 1 dog, and 1 cat were collected from 2013 to 2019 for viral isolation and detection of the presence of the gD gene by PCR. For sequence analysis, the gC partial gene was amplified. Results: Five strains were isolated from the dog, cat, and swine samples. The new PRV strains identified were confirmed by BLAST analysis, which revealed between 99.74% and 100% of similarity to the NIA-3 strain and phylogenetic analysis of the partial gene encoding the gC protein revealed that the PRV strains have divided into two main clades, clade 1 and clade 2. Conclusion: This report informed that most new cases of PRV were detected in the central regions of Argentina, where pig production is concentrated. The study in Bahía de Samborombón revealed a high percentage of detection but, the sampling is not representative of that of the rest of the country. Therefore, a systematic sampling effort of wild boar throughout the country should be included in the national program control. Although in Argentina only the inactivated Bartha vaccine is allowed, recombination risk should not be ignored if attenuated vaccines are incorporated into the National control plan. The two strains, one from the cat and one from the dog sample, are directly related to infected swine. The information about clinical cases and molecular characterization of new strains is important for a better understanding of the dynamics of PRV and to promote preventive measures.

Isolation and identification of two novel pseudorabies viruses with natural recombination or TK gene deletion in China.

Pseudorabies virus (PRV), the causative agent of Aujeszky's disease, has gained increased attention in China in recent years due to outbreaks of emergent pseudorabies. However, there is limited information about the evolution and pathogenicity of emergent PRV field strains in China. In this study, two PRV field strains were isolated from an intensive pig farm with suspected PRV infection. These were named the GXLB-2015 and GXGG-2016 strains and their growth characteristics together with their genome sequences and pathogenicity were determined. Nucleotide homology and phylogenetic analysis revealed the GXLB-2015 stain was relatively close to the foreign PRV isolated strains with respect to the whole genome sequence. However, it formed an independent branch between the foreign PRV isolates and the previous PRV variants isolated in China. Further recombination and genetic evolution analysis showed that the GXLB-2015 strain was a natural recombinant between the Bartha strain and PRV variants. The GXGG-2016 strain was highly homologous with the Chinese classical strains, but it has a natural deletion of 69 aa in the thymidine kinase (TK) gene. Pathogenicity analysis showed that, the GXLB-2015 strain had the strongest pathogenicity to mice with an LD50 of 103.5, while the GXGG-2016 strain with the TK gene deletion was not pathogenic to mice. Taken together, our data provide direct evidence for the genomic recombination and natural TK gene deletion of PRVs, which may provide a reference for a better understanding of PRV evolution in China and contribute to the clinical control of PRV infection in pig farms.

Epidemiology of Aujeszky disease in wild boars (Sus scrofa L.) in Croatia.

Aujeszky disease (AD) or pseudorabies is a viral disease of domestic and wild animals caused by the Suid alphaherpesvirus 1. In wild boar infection usually undergo latent phase but under certain conditions reactivation of the virus can result in a disease. Seroprevalence in wild boars ranges from 0.8 to 100%, and is among other influenced by region, type of management, age and sex of the studied animals. In this study we analyzed blood, lungs, olfactory bulbs and spleen from 222 free-living wild boars from different localities in Croatia and compared results obtained by ELISA with PCR, sex, age and locality. Total seroprevalence was 33.78%, ranging from 25.26% in males to 40.15% in females (p = 0.0346; χ2 = 4.47). According to the age categories prevalence was 10% in offspring, 27.53% in subadults, and 66.75% in adults. Seroprevalence in adult males (66.66%) and females (65.30%) was almost identical. In males, significantly lower seroprevalence was detected in offspring compared to subadults (χ2 = 4.07, p < 0.05) and adults (χ2 = 31.04; p < 0.05), and in subadults compared to adults (χ2 = 15.13; p < 0.0001). Among females, adults had a significantly higher prevalence compared to offspring (χ2 = 19.27; p < 0.0001) and subadults (χ2 = 8.62; p < 0.01). Analysis between counties revealed Sisacko-moslavacka county as a hot-spot for AD. None of the samples was positive for ADV antigens. The observed trend in prevalence points to the fact that the main transmission occurs during one part of the year (most probably the mating season). Also, triggers for virus reactivation might be more complex than previously thought, since none of our samples, collected during the mating and hunting season, was PCR positive. Finally, we can conclude that adult males represent the main transmission link between different wild boar groups.

Pseudorabies Virus Associations in Wild Animals: Review of Potential Reservoirs for Cross-Host Transmission.

Pseudorabies virus (PRV) has received widespread attention for its potential health effects on humans, wildlife, domestic animals, and livestock. In this review, we focus on PRV dynamics in wildlife, given the importance of wild-origin PRV transmission to domestic and farm animals. Wild boars, pigs, and raccoons can serve as reservoirs of PRV, with viral transmission to domestic livestock occurring via several routes, such as wild herd exposure, contaminated meat consumption, and insect vector transmission. Many endangered feline and canine species can be infected with PRV, with acute disease and death within 48 h. The first confirmed human case of PRV infection in mainland China was reported in 2017. Thus, PRV exhibits potentially dangerous cross-host transmission, which is likely associated with inappropriate vaccination, poor awareness, and insufficient biosecurity. Currently, no vaccine provides full protection against PRV in all animals. Here, we summarize the epidemiology and pathogenesis of PRV infection in wild, domestic, and farmed animals, which may facilitate the design of novel therapeutics and strategies for controlling PRV infection and improving wildlife protection in China.

The Epidemiology and Variation in Pseudorabies Virus: A Continuing Challenge to Pigs and Humans.

Pseudorabies virus (PRV) can infect most mammals and is well known for causing substantial economic losses in the pig industry. In addition to pigs, PRV infection usually leads to severe itching, central nervous system dysfunction, and 100% mortality in its non-natural hosts. It should be noted that increasing human cases of PRV infection have been reported in China since 2017, and these patients have generally suffered from nervous system damage and even death. Here, we reviewed the current prevalence and variation in PRV worldwide as well as the PRV-caused infections in animals and humans, and briefly summarized the vaccines and diagnostic methods used for pseudorabies control. Most countries, including China, have control programs in place for pseudorabies in domestic pigs, and thus, the disease is on the decline; however, PRV is still globally epizootic and an important pathogen for pigs. In countries where pseudorabies in domestic pigs have already been eliminated, the risk of PRV transmission by infected wild animals should be estimated and prevented. As a member of the alphaherpesviruses, PRV showed protein-coding variation that was relatively higher than that of herpes simplex virus-1 (HSV-1) and varicella-zoster virus (VZV), and its evolution was mainly contributed to by the frequent recombination observed between different genotypes or within the clade. Recombination events have promoted the generation of new variants, such as the variant strains resulting in the outbreak of pseudorabies in pigs in China, 2011. There have been 25 cases of PRV infections in humans reported in China since 2017, and they were considered to be infected by PRV variant strains. Although PRV infections have been sporadically reported in humans, their causal association remains to be determined. This review provided the latest epidemiological information on PRV for the better understanding, prevention, and treatment of pseudorabies.

Epidemiological Investigation and Genetic Analysis of Pseudorabies Virus in Yunnan Province of China from 2017 to 2021.

In recent years, the prevalence of pseudorabies virus (PRV) has caused huge economic losses to the Chinese pig industry. Meanwhile, PRV infection in humans also sounded the alarm about its cross-species transmission from pigs to humans. To study the regional PRV epidemic, serological and epidemiological investigations of PRV in pig populations from Yunnan Province during 2017-2021 were performed. The results showed that 31.37% (6324/20,158, 95% CI 30.73-32.01) of serum samples were positive for PRV glycoprotein E (gE)-specific antibodies via enzyme-linked immunosorbent assay (ELISA). The risk factors, including the breeding scale and development stage, were significantly associated with PRV seroprevalence among pigs in Yunnan Province. Of the 416 tissue samples collected from PRV-suspected pigs in Yunnan Province, 43 (10.33%, 95% CI 7.41-13.26) samples were positive for PRV-gE nucleic acid in which 15 novel PRV strains from these PRV-positive samples were isolated, whose gC and gE sequences were analyzed. Phylogenetic analysis showed that all 15 isolates obtained in this study belonged to the genotype II. Additionally, the gC gene of one isolate (YuN-YL-2017) was genetically closer to variant PRV strains compared with others, while the gE gene was in the same clade with other classical PRV strains, indicating that this isolate might be a recombinant strain generated from the classical and variant strains. The results revealed the severe PRV epidemic in Yunnan Province and indicated that PRV variants are the major genotypes threatening the pig industry development.

The Genetic Characterization of a Novel Natural Recombinant Pseudorabies Virus in China.

We sequenced the complete genome of the pseudorabies virus (PRV) FJ epidemic strain, and we studied the characteristics and the differences compared with the classical Chinese strain and that of other countries. Third-generation sequencing and second-generation sequencing technology were used to construct, sequence, and annotate an efficient, accurate PRV library. The complete FJ genome was 143,703 bp, the G+C content was 73.67%, and it encoded a total of 70 genes. The genetic evolution of the complete genome and some key gene sequences of the FJ strain and PRV reference strains were analyzed by the maximum likelihood (ML) method of MEGA 7.0 software. According to the ML tree based on the full-length genome sequences, PRV FJ strain was assigned to the branch of genotype II, and it showed a close evolutionary relationship with PRV epidemic variants isolated in China after 2011. The gB, gC, gD, gH, gL, gM, gN, TK, gI, and PK genes of the FJ strain were assigned to the same branch with other Chinese epidemic mutants; its gG gene was assigned to the same branch with the classic Chinese Fa and Ea strains; and its gE gene was assigned to a relatively independent branch. Potential recombination events were predicted by the RDP4 software, which showed that the predicted recombination sites were between 1694 and 1936 bp, 101,113 and 102,660 bp, and 107,964 and 111,481 bp in the non-coding region. This result broke the previously reported general rule that pseudorabies virus recombination events occur in the gene coding region. The major backbone strain of the recombination event was HLJ8 and the minor backbone strain was Ea. Our results allowed us to track and to grasp the recent molecular epidemiological changes of PRV. They also provide background materials for the development of new PRV vaccines, and they lay a foundation for further study of PRV.

Whole-Genome Sequence Analysis of Pseudorabies Virus Clinical Isolates from Pigs in China between 2012 and 2017 in China.

Pseudorabies virus (PRV) is an economically significant swine infectious agent. A PRV outbreak took place in China in 2011 with novel virulent variants. Although the association of viral genomic variability with pathogenicity is not fully confirmed, the knowledge concerning PRV genomic diversity and evolution is still limited. Here, we sequenced 54 genomes of novel PRV variants isolated in China from 2012 to 2017. Phylogenetic analysis revealed that China strains and US/Europe strains were classified into two separate genotypes. PRV strains isolated from 2012 to 2017 in China are highly related to each other and genetically close to classic China strains such as Ea, Fa, and SC. RDP analysis revealed 23 recombination events within novel PRV variants, indicating that recombination contributes significantly to the viral evolution. The selection pressure analysis indicated that most ORFs were under evolutionary constraint, and 19 amino acid residue sites in 15 ORFs were identified under positive selection. Additionally, 37 unique mutations were identified in 19 ORFs, which distinguish the novel variants from classic strains. Overall, our study suggested that novel PRV variants might evolve from classical PRV strains through point mutation and recombination mechanisms.

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.

Spatial variation in direct and indirect contact rates at the wildlife-livestock interface for informing disease management.

Little is known about disease transmission relevant contact rates at the wildlife-livestock interface and the factors shaping them. Indirect contact via shared resources is thought to be important but remains unquantified in most systems, making it challenging to evaluate the impact of livestock management practices on contact networks. Free-ranging wild pigs (Sus scrofa) in North America are an invasive, socially-structured species with an expanding distribution that pose a threat to livestock health given their potential to transmit numerous livestock diseases, such as pseudorabies, brucellosis, trichinellosis, and echinococcosis, among many others. Our objective in this study was to quantify the spatial variations in direct and indirect contact rates among wild pigs and cattle on a commercial cow-calf operation in Florida, USA. Using GPS data from 20 wild pigs and 11 cattle and a continuous-time movement model, we extracted three types of spatial contacts between wild pigs and cattle, including direct contact, indirect contact in the pastoral environment (unknown naturally occurring resources), and indirect contact via anthropogenic cattle resources (feed supplements and water supply troughs). We examined the effects of sex, spatial proximity, and cattle supplement availability on contact rates at the species level and characterized wild pig usage of cattle supplements. Our results suggested daily pig-cattle direct contacts occurred only occasionally, while a significant number of pig-cattle indirect contacts occurred via natural resources distributed heterogeneously across the landscape. At cattle supplements, more indirect contacts occurred at liquid molasses than water troughs or molasses-mineral block tubs due to higher visitation rates by wild pigs. Our results can be directly used for parameterizing epidemiological models to inform risk assessment and optimal control strategies for controlling transmission of shared diseases.

AUJESZKY'S DISEASE IN HUNTED WILD BOAR (SUS SCROFA) IN THE IBERIAN PENINSULA.

Aujeszky's disease (AD, pseudorabies) eradication programs in domestic pigs are implemented in several European countries where AD virus (ADV) circulates in local wild boar (Sus scrofa), making studies on ADV infection dynamics in wild boar increasingly relevant. The objective of our study was to characterize ADV dynamics in wild boar at a site in central Portugal and compare this site to three enzootic sites in central Spain. A total of 235 wild boar were sampled during the hunting season 2014-15. We collected serum, tissues (oropharyngeal tonsils and trigeminal and sacral ganglia), and swabs (oral, nasal, and genital) and analyzed these samples to detect ADV antibodies (enzyme-linked immunosorbent assay) and DNA (PCR). An overall seroprevalence of 42.6% was found (range 12.7-57.7%), being highest in adults (54.1%; 72/133). Overall, 2.8% (3/108) oral, 6.4% (7/109) nasal, and 12.8% (12/94) genital swabs were PCR positive. We found 20.4% (20/98) of the wild boar had at least one positive swab and were considered shedders. We found ADV in tissues of five animals; of 111 tonsils, three (2.7%) were PCR positive. Trigeminal (2/48; 4%) and sacral (2/53; 4%) ganglia collected in central Portugal, pertaining to three animals, were positive for ADV DNA. Logistic regression models showed that seroprevalence was influenced by site and age, whereas ADV shedding was influenced by site. Our study describes patterns of ADV infection in wild boar in Portugal and shows that wild boar also pose a risk, albeit lower than that in central Spain, for the eradication of AD from extensively managed domestic pigs in Portugal.

Epizootic situation of Aujeszky's disease within the territory of the Republic of Kazakhstan.

Background: The relevance of the study is determined by the danger that this disease poses to all types of domestic animals in the Republic of Kazakhstan, as well as the significant damage caused by this situation to the entire agriculture of the Republic of Kazakhstan. Timely and competent analysis of the epizootic situation for this disease in the country will reliably determine the development tendencies of this condition and predetermine a set of measures that should be taken to combat this animal disease in agricultural regions of the Republic of Kazakhstan. Aim: This study examines the development of the epizootic situation of Aujeszky's disease within the territory of the Republic of Kazakhstan. Methods: To achieve the set research goal, a combination of quantitative and qualitative research methods are used. Results: The results of this research article can be of significant importance for livestock breeders in the Republic of Kazakhstan, who are working on raising the number of pigs and other types of domestic animals and are interested in a further increase in the number. Conclusion: From a practical standpoint, the significance of this study is conditioned by the possibility of searching and determining the main directions of the spread of the disease under consideration in agricultural lands and regions of the Republic of Kazakhstan, as well as taking a set of measures to eradicate such a disease and prevent its future occurrence in the country.

Seroprevalence investigation and genetic analysis of pseudorabies virus within pig populations in Henan province of China during 2018-2019.

In late 2011, the outbreak of pseudorabies (PR) occurred in Bartha-K61-vaccinated pig farms and spread rapidly to many provinces of China, causing substantial economic losses to the swine industry. A total of 4708 pig serum samples from Henan province during 2018-2019 were collected to screen for the presence of pseudorabies virus (PRV) gE-specific antibodies, and phylogenetic analysis based on the gE gene of PRV was performed. Of the 4708 serum samples tested, 30.14% (1419/4708) were seropositive for PRV antibodies, based on PRV gE-coated enzyme-linked immunosorbent assay (ELISA), with slaughterhouses having the highest seroprevalence. The seropositive rates of PRV also varied with the region and the season. Phylogenetic analysis showed that three PRV isolates from this study were clustered in an independent branch together with the Chinese variant PRV strains (after 2012), and had a closer genetic relationship with the Chinese variant PRV strains, but differed genetically from the 4 early Chinese PRV strains and 4 European-American strains. This study suggests that three PRV isolates may belong to PRV variants, and the development of a novel vaccine against PRV variants is particularly urgent.

Pseudorabies virus infection in hunting dogs in Oita, Japan: Report from a prefecture free from Aujeszky's disease in domestic pigs.

We isolated two pseudorabies virus (PRV) isolates (designated OT-1 and OT-2) from two hunting dogs exhibiting neurological manifestations after eating the flesh of wild boar hunted in Oita prefecture, Kyushu Island, Japan. The isolates corresponded to a previously reported PRV (MY-1 strain) isolated from a hunting dog in neighboring Miyazaki prefecture, and it clustered into genotype II based on the glycoprotein C sequence. Our results suggest that this common PRV strain may have been maintained in wild boars on Kyushu Island even though domestic pigs in this area have attained an Aujeszky's disease-free status.

Current Status and Challenge of Pseudorabies Virus Infection in China.

Pseudorabies (PR), also called Aujeszky's disease, is a highly infectious disease caused by pseudorabies virus (PRV). Without specific host tropism, PRV can infect a wide variety of mammals, including pig, sheep, cattle, etc., thereby causing severe clinical symptoms and acute death. PRV was firstly reported in China in 1950s, while outbreaks of emerging PRV variants have been documented in partial regions since 2011, leading to significant economic losses in swine industry. Although scientists have been devoting to the design of diagnostic approaches and the development of vaccines during the past years, PR remains a vital infectious disease widely prevalent in Chinese pig industry. Especially, its potential threat to human health has also attracted the worldwide attention. In this review, we will provide a summary of current understanding of PRV in China, mainly focusing on PRV history, the existing diagnosis methods, PRV prevalence in pig population and other susceptible mammals, molecular characteristics, and the available vaccines against its infection. Additionally, promising agents including traditional Chinese herbal medicines and novel inhibitors that may be employed to treat this viral infection, are also discussed.