Molecular Detection of Porcine Cytomegalovirus, Porcine Parvovirus, Aujeszky Disease Virus and Porcine Reproductive and Respiratory Syndrome Virus in Wild Boars Hunted in Serbia during 2023.

Porcine cytomegalovirus (PCMV) infection is widespread worldwide and has a high prevalence in swine herds, especially in countries with intensive swine production. PCMV is zoonotic and can impact xenotransplants. It is the third swine virus known to be zoonotic, following swine influenza virus (influenza A) and hepatitis E virus genotype 3 (HEVgt3 or HEV-3). Wild boars, serving as reservoirs for various pathogens, including PCMV, pose a risk to both the pig industry and public health. This study aimed to investigate PCMV infection in Serbian wild boars using real-time PCR and assess other viral infections. We also tested samples for the presence of other viral infections: Aujeszky disease virus (ADV), Porcine parvovirus (PPV) and Porcine reproductive respiratory syndrome (PRRSV). Samples from 50 wild boars across 3 districts were tested. Results showed 8% positivity for PCMV DNA, with females showing higher infection rates. Porcine parvovirus (PPV) was detected in 56% of samples, while Porcine reproductive respiratory syndrome virus (PRRSV) was absent. ADV was found in 18% of samples, primarily in younger animals. This research contributes to understanding PCMV prevalence in Serbian wild boars and emphasizes the importance of monitoring viral infections in wild populations, considering the potential zoonotic and economic implications.

Seroprevalence of Viral Enzootic Diseases in Swine Backyard Farms in Serbia.

Contrary to pig farming in developed Western countries, in a large part of the world, pigs are still traditionally kept in small backyard farms, usually for family needs. Their main characteristics are low biosecurity, swill feeding, natural breeding and uncontrolled trade. Given the high number of backyard farms in Serbia and the risk they are thought to pose to intensive pig farming, the main aim of this study was to evaluate the prevalence of major viral diseases of swine among traditionally kept pigs in small holdings with low biosecurity. For this investigation, 222 serum samples from 69 backyard holdings were randomly selected and tested for antibodies to Porcine Reproductive and Respiratory Syndrome virus (PRRSV), Aujeszky's disease virus (ADV), Porcine Parvovirus (PPV) and Swine influenza Virus (SIV) by enzyme-linked immunosorbent assay (ELISA). The herd-level seroprevalence of PRRS, Aujeszky's disease and PPV was 2.9%, 27.5% and 37.7%, respectively. Swine influenza seroconversion was not confirmed in any of the tested holdings. Despite widely distributed PPV and AD in backyard farms in Serbia, almost 50% of them are still negative for all the tested diseases. The backyard farms must be monitored, and owners must be educated as their role in eradication programs and obtaining country-free status may be crucial.

A Salmonella enterica subspecies enterica serovar Choleraesuis outbreak in weaned piglets in Serbia: clinical signs, pathologic changes, and microbiologic features.

Salmonella enterica subspecies enterica serovar Choleraesuis is rarely detected in Europe, but the clinical disease has been reported in wild boars. We describe here the clinical findings, pathologic changes, and microbiologic features of swine salmonellosis caused by S. enterica serovar Choleraesuis in weaned piglets in Serbia. In April 2019, on a large farrow-to-finish pig farm, increased mortality was reported in weaned piglets, marked by lethargy, anorexia, pyrexia, and respiratory distress. Gross pathology revealed dermal cyanosis, mesenteric lymphadenopathy, splenomegaly, hepatomegaly, interstitial pneumonia, and colitis. By direct culturing of lung, liver, spleen, and lymph nodes, S. enterica ser. Choleraesuis variant Kunzendorf was isolated after years of absence of the disease in pig farms in Europe. The source of this salmonellosis outbreak caused by S. enterica ser. Choleraesuis remains unknown.

Investigation of possible aetiological/triggering factors in porcine ear necrosis syndrome at a farrow-to-feeder pig system.

At a commercial farrow-to-feeder pig system with 2,100 sows in Serbia, lesions resembling porcine ear necrosis syndrome were observed in 80% of the weaned pigs at 45-50 days of age. Pathomorphological examinations were carried out on 10 pigs that had been found dead. The gross lesions ranged from mild, superficial dermatitis to severe, deep inflammation with exudation, ulceration and necrosis. Histopathological examination revealed erosive and ulcerative dermatitis of the pinna with neutrophilic and lymphocytic infiltration and bacterial colonies in the crusts. Staphylococcus aureus (MRSA strain), Staphylococcus hyicus and Streptococcus group C were cultivated from eight, S. hyicus from two ear tissue scraping samples. All 10 samples were positive for treponemes and phylogenetic analysis of two polymerase chain reaction products confirmed the relationship to Treponema (T.) medium/vincentii and Treponema pedis. Treponemes were also detected in seven oral swabs that were analysed to obtain evidence of the transmission of this bacterium by ear biting. The contribution of non-infectious factors to this misbehaviour could not be ruled out as the crude protein concentration of the feed was inappropriate and the climate of the pig house was suboptimal. The concentrations of selected mycotoxins in the feed were not elevated. However, the contribution of both infectious and non-infectious factors to the onset of disease was most probable.

Morphological and immunophenotypic characteristics of the liver of swine naturally infected with hepatitis E virus.

Hepatitis E virus (HEV), the zoonotic agent of infectious hepatitis, is present in swine farms in different geographical areas. Little is known about the mechanism of liver damage and type of local immune response by HEV in swine. Therefore, the aim of this study was to determine the morphological and immunophenotypic characteristics of hepatic lesions caused by hepatitis E virus in naturally infected swine. In this study, liver samples of 12 slaughtered 10 weeks old pigs which were RT-PCR positive for HEV RNA in rectal swab samples have been used. Livers were macroscopically examined and samples were taken for histopathological, immunohistochemical (CD3, CD79α and TGF-ß1), semiquantitative, morphometric analysis, RT-nested-PCR, PCR and bacteriological analysis. Microscopically, mild and moderate multifocal lymphoplasmacytic hepatitis was observed. Apoptotic bodies were observed as areas of focal eosinophilic condensation in the cytoplasm of 33.33% liver samples, while in 16.67% liver samples portal fibrosis was detected. Immunohistochemically, portal and lobular lymphocytes in the mononuclear liver infiltrate were predominantly CD3+ T cells (234.80 ± 79.98). An intense TGF-ß1 positive reaction was observed within the mononuclear cell infiltrate as well as polymorphonuclear cells in liver samples with apoptosis of hepatocytes. In all 12 tested liver samples HEV RNA was detected by RT-nested-PCR. HEV is noncytopathic, and this finding provides further evidence for an immune mediated pathogenesis in hepatitis E virus infection in swine. Also, the role of CD3+ cells in hepatocyte damage is clearly demonstrated.

Seroprevalence of hepatitis E in pigs and wild boars in the region of the city Belgrade.

INTRODUCTION: Hepatitis E is considered an emerging human viral disease with many evidences of zoonotic nature of disease, and swine are the main reservoir of HEV. The aim of this study was to determine HEV seroprevalence in commercial pig farms, backyard pigs, slaughtered pigs and wild boars in the region of the city Belgrade. METHODOLOGY: A total of 405 sera samples: 150 samples from 3 commercial pig farms, 70 samples from backyard pigs, 119 samples from slaughtered pigs and 66 samples from wild boars of the region of the city Belgrade, Serbia were analysed by commercial ELISA test. RESULTS: The overall HEV seroprevalence in 3 commercial pig farms was 55.33% (83/150). All tested farms (farm A, B and C) were positive on the presence of anti-HEV antibodies, respectively 58% (29/50), 54% (27/50) and 54% (27/50). From 70 tested backyard pigs, 75.71% (53/70) were tested seropositive. In total, 26 backyard pig holidngs were confirmed as positive to anti-HEV antibodies (81.25%). At slaughterhouse, 25% (8/32) weaned piglets and 20.69% (18/87) fattening pigs were tested positive on anti-HEV antibodies. Overall HEV seroprevalence in tested wild boar population was 52.25% (36/66). CONCLUSIONS: Detected very high seroprevalence of anti-HEV antibodies indicated an active circulation of HEV, being enzootic in the swine population, and wild boars, as well, in the region of the city Belgrade.

Bovine viral diarrhea virus infection in wild boar.

Bovine viral diarrhea (BVD) is one of the most economically important diseases of cattle. With its very high prevalence, cattle kept on pastures become a source of the virus for the wildlife which, due to their susceptibility, then easily can serve as a source for re-infections of cattle. Therefore, we investigated the BVDV infection in Serbian wild boar and assessed the role of wild boar in BVDV epidemiology including possible spreading to domestic species. This study was based on examination of 50 spleen samples which were collected from wild boars located in Eastern Serbia during the hunting season 2016/2017. BVDV genome was detected in 4 of 50 samples (8%). Phylogenetic analysis based on 5'UTR revealed that BVDV strains from wild boars shared 100% identity. Belonging to the BVDV 1f subgenotype, the most common in cattle, we showed that BVDV infections of wild boar occurred as a result of either direct or indirect contact with domestic animals. Therefore, the occurrence of infectious disease in wildlife emphasizes the need to study the pathogens shared by wildlife and domestic animals by investigating the incidence of pathogens and disease patterns of those populations.