The first case of African swine fever in domestic pigs in Serbia was in 2019. The following year, the disease was confirmed in wild boar. Thenceforth, ASF has been continuously reported in both wild and domestic pigs. The outbreaks in domestic pigs could not be linked directly to wild boars, even though wild boars were endemically infected, and reservoirs for ASF. This study aimed to investigate outbreaks and routes of transmission in domestic pigs in a region of central Serbia where no outbreaks in wild boar were reported. Fourteen outbreaks of ASF on backyard farms with low biosecurity were traced back, and no connection to wild boar was found. The epidemic investigation covered 2094 holdings, with 24,368 pigs, out of which 1882 were tested for ASF. In surrounding hunting grounds, field searches were conducted. Dead wild boars were found, and 138 hunted wild boars were negative for ASFV. It was concluded that outbreaks in 2021 were provoked by the illegal trade of live animals and pig products. Even though infective pressure from wild boars is assumed, no positive cases have been found, while the ASFV spreads within the domestic swine population evidenced in four recent outbreaks in 2022.

Introduction: West Nile Virus, an emerging zoonotic pathogen, has been circulating in Serbia for over a decade, with its first detection in mosquitoes in 2010. Since then, the virus has led to increasing cases in both animals and humans, peaking in 2018 with 415 human cases and 36 fatalities. This study aimed to explore the phylogenetic relationships between previously sequenced West Nile virus strains from Serbia and those sequenced in this study, while also identifying possible virulence factors.Materials and methods: Whole genome sequencing was conducted using a targeted approach on the MinION Mk1C platform, following a two-step process involving cDNA synthesis and amplification. Bioinformatics analysis included demultiplexing, primer trimming, and sequence mapping using tools such as iVar, Minimap2, and Samtools. Phylogenetic analysis was performed using MAFFT alignment and the Maximum Likelihood method with the Tamura Nei model in MEGA X software. Virulence factors were assessed in both structural and nonstructural proteins, focusing on key glycosylation motifs and specific mutations. Homology modeling of the E protein was also performed to evaluate potential structural changes due to mutations.Results: Phylogenetic analysis revealed two major sublineages within the E subclade, representing the majority of strains from Western and Central Europe. These sublineages likely originated from Austria, Serbia, and Hungary between 2008 and 2012. The study also identified three distinct sublineages within the D subclade, which includes more diverse strains from Southern Europe. The E protein exhibited significant variations, particularly at the E159 site, which is crucial for virulence. The EI159T aa change has become dominant in recent years, replacing the previously prevalent EI159M. Additionally, changes in the NS1 glycoprotein and NS3 protein, both of which are involved in immune modulation and viral replication, were identified, with potential implications for the virus’s virulence.Conclusion: The study’s findings highlight the Western Balkans and Central Europe as key regions for the mixing and dissemination of West Nile virus strains from both Western-Central and Southern Europe. These results underscore the importance of continuous surveillance and phylogenetic analysis to monitor the evolution and spread of West Nile virus, particularly in light of the frequent mutations observed in virulence-associated sites.

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.

Canine distemper virus (CDV) is a highly contagious and often fatal disease affecting wild and domesticated carnivores. The virus is a single-stranded RNA virus from the genus Morbillivirus and the family Paramyxoviridae. While domestic dogs are the most common hosts, the virus poses a significant threat to endangered wildlife due to its broad host range. This study aimed to characterize the CDV Haemagglutinin (H) gene in golden jackals and explore the molecular evolution of the virus in an underrepresented host. A total of 88 brain samples from hunted golden jackals were tested for the presence of CDV viral nucleic acid, and the H gene of positive samples was amplified and sequenced using the Sanger method. Phylogenetic analysis, conducted using maximum likelihood methods, revealed that all Serbian sequences clustered within the Arctic lineage. Notably, the analysis identified a tyrosine (Y) at position 549 of the H protein, a mutation commonly associated with wildlife hosts, instead of the histidine (H) typically found in domestic strains. Additionally, a mutation at position 310 was observed, which could potentially affect the protein’s function and virus-host interactions. These findings provide valuable insights into the genetic diversity and evolutionary dynamics of CDV in golden jackals, with broader implications for understanding the virus’s adaptability to different hosts. Further research is needed to investigate the functional impact of these mutations, particularly their role in vaccine efficacy and disease transmission across wildlife and domestic species.