Implementation and monitoring of oral rabies vaccination of foxes in Kosovo between 2010 and 2013--an international and intersectorial effort.

The European Union has used instrument for pre-accession (IPA) funds to provide technical assistance and supplies for the eradication, monitoring and control of rabies in several pre-accession countries. As a result, since 2010, multi-annual oral rabies vaccination (ORV) programmes for eliminating fox rabies have been launched in six Western Balkan countries. Here the implementation of the ORV programme in Kosovo, the smallest of the West Balkan countries, is described. Associated challenges under difficult political conditions, potential biases, and the results of rabies surveillance and monitoring of ORV campaigns (bait uptake and immunisation rates) since 2010 are reported.

Emergence and Persistent Circulation of Highly Pathogenic Avian Influenza Virus A (H5N8) in Kosovo, May 2021-May 2022.

In this study, we report the first outbreak of highly pathogenic avian influenza (HPAI) A H5N8, clade 2.3.4.4b in Kosovo on 19 May 2021. The outbreak consisted of three phases: May-June 2021, September-November 2021, and January-May 2022. In total, 32 backyards and 10 commercial holdings tested positive for the virus. Interestingly, the third and last phase of the outbreak coincided with the massive H5N1 clade 2.3.4.4b epidemic in Europe. Phylogenetic analyses of 28 viral strains from Kosovo revealed that they were closely related to the H5N8 clade 2.3.4.4.b viruses that had been circulating in Albania, Bulgaria, Croatia, Hungary, and Russia in early 2021. Whole genome sequencing of the 25 and partial sequencing of three H5N8 viruses from Kosovo showed high nucleotide identity, forming a distinctive cluster and suggesting a single introduction. The results of the network analysis were in accordance with the three epidemic waves and suggested that the viral diffusion could have been caused by secondary spreads among farms and/or different introductions of the same virus from wild birds. The persistent circulation of the same virus over a one-year period highlights the potential risk of the virus becoming endemic, especially in settings with non-adequate biosecurity.

A case report of sporadic ovine listerial meningoencephalitis in Kosovo.

In 2018, a case of neural disease suspected of listeriosis was reported in a flock of sheep in Kosovo with the death of ewes and 5 lambs. Samples from the brain of only three dead animals were subjected to histopathological and bacteriological analysis. MALDI-TOF MS was applied to confirm suspected Listeria spp. isolates from culture and multiplex PCR was applied for molecular serotyping. All isolates were tested for antimicrobial susceptibility by microdilution broth method. The histopathological analysis of the brain specimens showed typical changes for Listeria monocytogenes. Listeria spp. was isolated in brain samples from all three animals, and all the isolates were confirmed using MALDI-TOF MS and PCR down to the species level (Listeria monocytogenes). The molecular characterisation using multiplex PCR revealed all isolates as Listeria monocytogenes serotype 4b. All L. monocytogenes isolates were found to be susceptible to penicillin, erythromycin, tetracycline, streptomycin, trimethoprim/sulfamethosazole, quinupristin/dalfopristin, kanamycin, vancomycin, and gentamicin but resistant to nitrofurantoin and lincomycin. This study shows the emergence of a highly virulent strain in sheep farms in Kosovo and a possible threat to public health.

Seasonal variation in bait uptake and seropositivity during a multi-year biannual oral rabies fox vaccination programme in Kosovo (2010-2015).

The European Union supported programmes for rabies control in Kosovo between 2010 and 2015, including spring and autumn biannual oral vaccination campaigns targeting foxes. Throughout the programmes foxes were obtained to provide samples for monitoring the campaigns. This paper explores the seasonal pattern of bait uptake and seropositivity in the fox population. Bait uptake varied by season and by the phase of the project supporting the programme (the main differences between phases being the number of baits distributed and flight line separation). Seropositivity varied by season and by titre of the vaccine used in the preceding campaign. The analyses also suggested a negative effect of higher daytime temperature on bait uptake, and possible association between geographic location of sampling and concordance between bait uptake and seropositivity, but the dataset was too unbalanced to support robust conclusions on these detailed aspects. Descriptive summaries of the data and the multilevel analyses showed that the proportion of sampled foxes that were positive for bait uptake and the proportion seropositive were both high through winter, following the autumn campaigns, and declined through spring and summer, with a low point in samples collected during the time when juvenile foxes are typically dispersing from their birth dens. The percentage of foxes positive for bait uptake was below 30 % (first project phase) and 40 % (second project phase) from mid-July to mid-October following a spring campaign, compared with around 70 % (first project phase) and 80 % (second project phase) in the periods between autumn and the following spring campaigns. As could be expected, the percent of samples that were seropositive followed a similar seasonal pattern, with some additional variation associated with the titre of vaccine used. This seasonal pattern is likely because the population sampled in the late summer months includes increasing numbers of young foxes that could not have been effectively exposed to the spring vaccination campaign, and would have lost any possible maternal immunity by late summer. The main finding of high levels of bait uptake and seroprevalence through winter, following the autumn campaigns, declining through summer despite the implementation of spring campaigns, supports advice that countries lacking financial resources to support biannual campaigns should focus resources on once per year vaccination in late autumn or early winter. This pattern also indicates that a fox population may rapidly become naïve after cessation of vaccination programmes, therefore strongly coordinated regional programmes and good surveillance will be needed.

Genotyping and phylogenetic analysis of bovine viral diarrhea virus (BVDV) isolates in Kosovo.

Three serum samples positive in Antigen ELISA BVDV have been tested to characterise genetic diversity of bovine viral diarrhea virus (BVDV) in Kosovo. Samples were obtained in 2011 from heifers and were amplified by reverse transcription-polymerase chain reaction, sequenced and analysed by computer-assisted phylogenetic analysis. Amplified products and nucleotide sequence showed that all 3 isolates belonged to BVDV 1 genotype and 1b sub genotype. These results enrich the extant knowledge of BVDV and represent the first documented data about Kosovo BVDV isolates.

Entomological research on the vectors of bluetongue disease and the monitoring of activity of Culicoides in the Prishtinë region of Kosova.

Clinical bluetongue (BT) caused by BT virus serotype 9 (BTV-9) was observed in Kosova in 2001 and, although subsequently no further clinical cases was diagnosed, its continuing presence has been demonstrated by serological tests in cattle, sheep and goats. In this study, light traps were placed in stables near Prishtinë to identify possible vectors of BTV in Kosova. Samples were collected from October 2004 until the end of 2006. Culicoides were identified and speciated and results were plotted against temperature data. Samples contained Obsoletus and Pulicaris Complexes but not C. imicola. The first specimens of Culicoides were collected in April and they continued to be detected until November. Generally, Obsoletus Complex was present in the largest numbers, with the exception of the middle of the year when the Pulicaris Complex predominated. The number of Culicoides trapped was directly linked to temperature (p<0.05) and records indicated that Culicoides activity ceased when minimum temperatures fell below 0°C; activity recommenced when minimum temperatures rose to approximately 6°C. These results indicate that there was a lack of a vector for BTV during winter for a period lasting approximately five months.