The results of the Serbian national integrated West Nile virus (WNV) surveillance program conducted in 2018 and funded by the Serbian Veterinary Directorate are presented. The WNV surveillance program encompassed the entire territory of Serbia and was conducted by the veterinary service in collaboration with entomologists and ornithologists. The objective of the program was early detection of WNV circulation in the environment and timely reporting to the public health service and local authorities to increase clinical and mosquito control preparedness. The program was based on the detection of WNV presence in wild birds (natural hosts) and mosquitoes (virus vectors) and on serological testing of sentinel horses (WNV-specific IgM antibodies). The season 2018 was confirmed to be the season of the most intensive WNV circulation with the highest number and severity of human cases in Serbia ever reported. The most intense WNV circulation was observed in the northern and central parts of Serbia including Vojvodina Province, the Belgrade City area, and surrounding districts, where most positive samples were detected among sentinel animals, mosquitoes and wild birds. The majority of human cases were preceded by the detection of WNV circulation during the surveillance. The WNV surveillance program in 2018 showed satisfactory results in the capacity to indicate the spatial distribution of the risk for humans and sensitivity to early detection of WNV circulation in the environment.

Equine arteritis virus (EAV) is responsible for infections in equids. It can spread easily within the horse population and has a major impact on the horse breeding industry. No EAV outbreak has ever been reported in Serbia. To determine whether EAV is nonetheless circulating there, especially in the Vojvodina region, 340 horse serum samples were subjected to serology testing to detect EAV antibodies. In parallel, semen samples from three seropositive stallions were collected to evaluate their EAV status, using RT-qPCR and virus isolation on cell culture. Horse sera with EAV antibodies represented 15.88% (54/340) of the tested samples, 83.23% (283/340) being negative, and just three samples (0.89%) being uninterpretable due to cytotoxicity. Only 7.2% (10/138) of horses kept by private owners on their own property were seropositive for EAV, whereas 21.8% (44/202) of horses kept on stud farms had EAV antibodies. Phylogenetic analysis showed that the Serbian EAV isolate was most closely related to isolates from the neighbouring Hungary. EAV is circulating in the Serbian horse population, especially among the breeding population certainly due to the use of EAV shedder stallions since there is no surveillance programme in Serbia and only limited checks on racehorses. Moreover, phylogenetic analysis indicates that the EAV isolated from a Lipizzaner stallion in Serbia is closely related to isolates from Hungary, and together form a new cluster.

During July 2018, the first clinical case of neurological West Nile virus (WNV) infection was reported in a Belgian sports mare in Belgrade, Serbia. Typical symptoms, such as hypersensitive skin reaction, disorientation, weakness, ataxia and the loss of equilibrium were reported. Detection of WNV IgM antibodies by commercial ELISA in the serum samples of the diseased mare strongly indicated acute infection. The ELISA positive results were confirmed by VNT. Hematological and biochemical parameters were in the reference range. The only finding was a minor lymphopenia. WNV RNA was not detected by RT-qPCR in the blood sample extracted seven days after the disease had broken out. The horse improved clinically in two weeks while other horses at the same premises remained asymptomatic. The clinical, serological, biochemical and molecular analyses applied confirmed the first clinical case of neuroinvasive WNV infection in horses in Serbia. The West Nile virus has been circulating in Serbia in the last decade in mosquitoes, birds, and horses, but no evidence of equine WNV clinical cases were registered so far.

The paper presents the findings of specific antibodies in the blood sera of donkeys against the following viruses: equine infectious anemia virus (EIAV), African horse sickness virus (AHSV), equine herpesvirus type 1 (EHV-1), equine influenza virus subtype H3N8 (EIV) and equine arteritis virus (EAV). The analyses were conducted during the year 2022. From a total of 199 donkeys bred in “Zasavica”, blood was sampled from 53 animals (2 male donkeys and 51 female donkeys), aged 3 to 10 years. Specific antibodies against EIAV were not detected in any of the tested animals using the agar-gel immunodiffusion (AGID) assay. No specific antibodies against AHSV, tested by enzyme-linked immunosorbent assay (ELISA), or antibodies against EAV, tested by the virus neutralization test (VNT) and ELISA were detected in any of these animals. A positive serological result for EHV-1 was determined by the VNT in all animals, with antibody titer values ranging from 1:2 to 1:128, while a very low antibody titer value for EIV (subtype H3N8) of 1:16 was determined in 18 donkeys using the hemagglutination inhibition test (HI test).

Hepatitis E virus (HEV) is the causative agent of acute hepatitis E infection in humans. Two epidemiological patterns of the disease exist—endemic and sporadic. Genotypes 1 (HEV-1) and 2 (HEV-2) are transmitted through contaminated water and are responsible for the outbreaks of many large-scale epidemics in developing countries of Asia and Africa. Genotypes 3 (HEV-3) and 4 (HEV-4) have zoonotic potential and cause sporadic cases and small outbreaks in high-income countries. The first case of HEV infection in swine was confirmed in 1997 and later detected in other animal species. The aim of this study was to investigate the seroprevalence of HEV infection in pig farms in Vojvodina province, Serbia. Three hundred blood samples were collected from five different categories of pigs from 3 different farms on the territory of the South Bačka district in Vojvodina (Serbia). The analyses were conducted by in-house ELISA, while the western blot method was used as a confirmatory test for doubtful results. The presence of HEV IgG was detected on all three examined farms. The established seroprevalence in Farm A was 37%, 31% in Farm B, and 54% in Farm C. The mean seroprevalence for all farms was 40.66%. A higher seroprevalence was found in fatteners compared to younger categories of pigs. We concluded that HEV is widespread on pig farms with intensive management. Further analyses should be conducted with the aim of implementing a surveillance program to prevent possible human infection.