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With their remarkable flight capabilities, wild and captive birds play a pivotal role in the global dissemination of zoonotic pathogens including , Avian Influenza viruses (AIV), Chikungunya virus (CHIKV), Usutu virus (USUV), and West Nile virus (WNV). They function both as hosts and reservoirs responsible for transporting the mentioned infectious agents across vast geographic regions. Additionally, captive birds and birds inhabiting urban environments, particularly in tourist destinations, present significant public health concerns due to facilitated close interactions with humans. A total of 358 samples originating from fifteen bird species were collected across 21 locations in Sarajevo Canton, over three consecutive years (2022-2024). Upon collection, the samples were subjected to molecular analysis to detect the presence of zoonotic pathogens. For detection of spp., and , real-time PCRs (qPCR) were used following established protocols. Additionally, reverse transcriptase real-time PCR (RT-qPCR) were utilized for the detection of emergent viral pathogens including avian influenza viruses, Chikungunya, Usutu, and West Nile virus. spp. was detected in 29.9% (95% CI: 25.2-34.9) of samples. Further, was identified in 10.3% (95% CI: 5.2-17.7) of positive samples originating from captive birds and birds inhabiting urban environments. One sample (0.3%) originating from a wild bird was positive to West Nile Virus. None of the samples tested positive for Avian Influenza viruses, Chikungunya and Usutu virus. The identification of and West Nile virus highlights the increased likelihood of zoonotic transmission. This underscores the imperative for bolstered biosecurity measures and public health strategies aimed at mitigating the risk associated with both environmental exposure and direct contact, especially in areas characterized by substantial tourist activity.

Influenza A virus continues to represent a growing problem affecting mainly birds but with an increasing number of mammal transmission events reported each year. Nevertheless, molecular characterization and phylogenetic analysis of influenza A viruses originating from all confirmed cases have not been systematically performed in all parts of the world. In this study, we investigated a sample originating from a mute swan that died in November 2021 in the northern part of Bosnia and Herzegovina with RT-qPCR and whole genome sequencing using ONT MinION. It was diagnosed as a highly pathogenic Influenza A virus, subtype H5N1 of 2.3.4.4b clade, and phylogenetic analysis revealed high sequence homology with other European HPAI H5N1 sequences at the time. The notable detected mutations in HA (N110S and T139P) and NA genes (H155Y), that facilitate the host specificity shift and enable the resistance to some antiviral drugs respectively, underscore the necessity of virus evolution surveillance. Therefore, the rapid dissemination of information, including virological and molecular data, is essential for the introduction of tailored prevention measures for infected animals, providing clearer insight and better awareness of a potential public health threat.

Bats are a natural host for a number of viruses, many of which are zoonotic and thus present a threat to human health. RNA viruses of the family Filoviridae, many of which cause disease in humans, have been associated with specific bat hosts. Lloviu virus is a Filovirus which has been connected to mass mortality events in Miniopterus schreibersii colonies in Spain and Hungary, and some studies have indicated its immense zoonotic potential. A die-off has been recorded among Miniopterus schreibersii in eastern Bosnia and Herzegovina for the first time, prompting the investigation to determine the causative agent. Bat carcasses were collected and subjected to pathological examination, after which the lung samples with notable histopathological changes, lung samples with no changes and guano were analyzed using metagenomic sequencing and RT-PCR. A partial Lloviu virus genome was sequenced from lung samples with histopathological changes and found to be closely related to Hungarian and Italian virus sequences. Further accumulation of mutations on the GP gene, coding the glycoprotein responsible for cell tropism and host preference, enhances the need for further characterization and monitoring of this virus to prevent spillover events and protect human health.

Rift Valley fever (RVF) is an arboviral zoonosis that primarily affects ruminants but can also cause illness in humans. The increasing impact of RVF in Africa and Middle East and the risk of expansion to other areas such as Europe, where competent mosquitos are already established, require the implementation of efficient surveillance programs in animal populations. For that, it is pivotal to regularly assess the performance of existing diagnostic tests and to evaluate the capacity of veterinary labs of endemic and non-endemic countries to detect the infection in an accurate and timely manner. In this context, the animal virology network of the MediLabSecure project organized between October 2016 and March 2017 an external quality assessment (EQA) to evaluate the RVF diagnostic capacities of beneficiary veterinary labs. This EQA was conceived as the last step of a training curriculum that included 2 diagnostic workshops that were organized by INIA-CISA (Spain) in 2015 and 2016. Seventeen veterinary diagnostic labs from 17 countries in the Mediterranean and Black Sea regions participated in this EQA. The exercise consisted of two panels of samples for molecular and serological detection of the virus. The laboratories were also provided with positive controls and all the kits and reagents necessary to perform the recommended diagnostic techniques. All the labs were able to apply the different protocols and to provide the results on time. The performance was good in the molecular panel with 70.6% of participants reporting 100% correct results, and excellent in the serological panel with 100% correct results reported by 94.1% of the labs. This EQA provided a good overview of the RVFV diagnostic capacities of the involved labs and demonstrated that most of them were able to correctly identify the virus genome and antibodies in different animal samples.

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which led to the COVID-19 pandemic, has significantly impacted global public health. The proper diagnosis of SARS-CoV-2 infection is essential for the effective control and management of the disease. This study investigated the SARS-CoV-2 infection using RT-qPCR tests from laboratories in Bosnia and Herzegovina. We performed a retrospective study of demographic data and Ct values from 170,828 RT-qPCR tests from April 2020 to April 2023, representing 9.3% of total national testing. Samples were collected from 83,413 individuals across different age groups. Of all tests, 33.4% were positive for SARS-CoV-2, with Ct values and positivity rates varying across demographics and epidemic waves. The distribution was skewed towards older age groups, although lower positivity rates were observed in younger age groups. Ct values, indicative of viral load, ranged from 12.5 to 38. Lower Ct values correlated with higher positive case numbers, while higher Ct values signaled outbreak resolution. Additionally, Ct values decreased during epidemic waves but increased with the dominance of certain variants. Ct value-distribution has changed over time, particularly after the introduction of SARS-CoV-2 variants of interest/concern. Established Ct value trends might, therefore, be used as an early indicator and additional tool for informed decisions by public health authorities in SARS-CoV-2 and future prospective pandemics. Moreover, they should not be overlooked in future epidemiological events.

This study aims to estimate the number of AMY2B gene copies and measure serum amylase activity in several Balkan dog breeds. Additionally, it explores the relationship between these genetic and biochemical parameters. Blood samples from 85 dogs representing eight breeds were collected, DNA was extracted, and AMY2B copy numbers were determined using droplet digital PCR. AMY2B gene copies ranged from 7.7 to 18.4, with a mean of 12.4 ± 2.2. Significant breed-related differences were observed (p = 0.025), with Istrian Wire-Haired Hounds showing the highest mean copy number (13.9 ± 1.5) and Posavatz Hounds the lowest (10.8 ± 1.5). Serum amylase activity ranged from 3.3 to 17.8 µkat/L, with a mean of 8.7 ± 2.6, and showed significant interbreed differences (p = 0.004), with Barak breed displaying the highest activity. Serum glucose levels varied widely, but no significant interbreed differences were detected (p = 0.340). No significant correlation was found between AMY2B copy numbers and serum amylase activity or glucose levels. The study concludes that Balkan dogs have AMY2B copy numbers similar to other European breeds, likely reflecting historical agricultural practices in the region, thereby facilitating better starch digestion. While significant variations exist among breeds, the lack of correlation between gene copy number and amylase activity suggests that other factors influence enzyme levels.

Background Crimean–Congo haemorrhagic fever (CCHF) is a widespread tick‐borne zoonosis with reported detection of virus and/or virus‐specific antibodies from over 57 countries across Africa, Asia, Europe and the Middle East and is endemic in the Balkans. Detection of Crimean–Congo Haemorrhagic Fever Virus (CCHFV) antibodies in domestic ruminants has been important in providing initial evidence of virus circulation and in localising CCHFV high‐risk spots for human infection. Objectives The present study investigated the possible exposure of sheep to CCHFV in Bosnia and Herzegovina (B&H). Methods To investigate the presence of anti‐CCHFV antibodies in sheep, all sera (n = 176) were tested using multi‐species double antigen enzyme‐linked immunosorbent assay (ELISA). Reactive sera were further complementary tested by adapted commercial indirect immunofluorescence assay (IFA) using FITC‐conjugated protein G instead of anti‐human immunoglobulins. Results CCHFV specific antibodies were detected in 17 (9.66%) animals using ELISA test. All negative sera were determined as negative by both tests, while 13 out of 17 ELISA‐positive reactors were also determined as unambiguously positive by IFA test. The age group with the highest proportion of seropositive rectors were the oldest animals. Conclusions This is the first report of anti‐CCHFV antibodies in sheep from B&H providing the evidence of CCHFV circulation in the country's sheep population. So far, these findings indicate the circulation of the virus in the westernmost region of the Balkans and point to the potential CCHFV spread further out of this endemic area. Crimean Congo Hemorrhagic Fever Virus (CCHFV) infection is endemic in Balkans. A serological survey of 176 sheep was conducted to investigate the circulation to CCHFV in Bosnia and Herzegovina. CCHFV specific antibodies were detected in 17 (9.66%) animals providing the early evidence of CCHFV circulation in the country's sheep population.

Background. The production of milk and dairy products and their placement on the market represent a constant profit for the farmers/producers in Bosnia and Herzegovina (BH). The profitable operation of the dairy farms is influenced by the reproductive performance of the lactating animals. This study assessed individual animal reproductive characteristics in selected dairy farms and described their reproductive performance indicators. Results. The median age at first insemination was 493 days (5th–95th percentile range 429–840), while the age at first calving was 802 days (5th–95th percentile range 708–1168). The median pregnancy proportion at first insemination was 40% (5th–95th percentile range 17–62), while the cumulative pregnancy rate calculated at day-60, day-80, day-100, and day-120 showed that approximately 64% of all pregnancies happened before day-120. The calculated interservice intervals showed that approximately 69% of the repeat breeding animals came back to the oestrus in the period of 18 to 24 days. This is an indication of very good oestrus detection in selected dairy farms. The mean number of services per pregnancy was 2.61 (range 1–12). The median calving-to-first-insemination interval was 62.5 days (5th–95th percentile range 16–408). The calving-to-conception interval was 101 day (5th–95th percentile range 36–506). Finally, the calving interval was 385 days (5th–95th percentile range 329–773). Conclusions. There is a need for an organised, regular, and more comprehensive recording system for the reproduction of dairy cattle among dairy farms in Una-Sana Canton. The calculated reproductive measures indicated an undulant trend in reproductive performance among selected dairy farms in Una-Sana Canton. Knowing the apparent reproductive indicators described in this study, the farmers and veterinary authorities may identify and correct areas in their management that contribute to the reproductive underperformance.

Anthelmintic resistance in livestock is a growing concern worldwide, with significant implications for animal health and agricultural productivity. This study explores the perceptions of veterinarians and farmers in Bosnia and Herzegovina regarding the factors contributing to anthelmintic resistance in Haemonchus contortus nematodes. Data were collected through structured questionnaires completed by 106 veterinarians and 188 farmers in 2022 and 2023. The analysis focused on self-reported therapeutic practices, farm management and environmental variables. Logistic regression, including Firth’s penalized approach, was used to assess associations between these perceived factors and the reported occurrence of resistance. Notably, combination anthelmintic treatments were perceived as a significant risk factor (OR > 49.3), while higher altitude was seen as potentially protective (OR = 0.10). Routine prophylactic deworming was associated with an increased likelihood of perceived resistance (OR = 173.7), whereas staying informed about newly registered products was perceived as protective (OR = 0.34). Although the findings are based on the self-reported perceptions and practices of veterinarians and farmers, they align with globally recognized trends and offer the first structured insights into factors perceived to contribute to anthelmintic resistance in Bosnia and Herzegovina. This study underscores the importance of awareness and responsible anthelmintic use and the need for improved diagnostics and ongoing education to combat anthelmintic resistance.

Gastrointestinal nematodes, particularly Haemonchus contortus, represent a major threat to ruminant health and productivity worldwide, largely due to the widespread emergence of anthelmintic resistance. In Bosnia and Herzegovina, benzimidazole resistance has previously been confirmed in domestic ruminants; however, data on wildlife remain lacking. Given the frequent spatial and temporal overlap between domestic and wild ruminants on shared pastures, this study aimed to investigate the occurrence of benzimidazole-resistant H. contortus genotypes within a multi-host system. During the 2024/2025 season, a total of 111 abomasal samples were collected from sheep (n = 20), lambs (n = 12), goats (n = 17), roe deer (n = 40) and chamois (n = 22) across four localities in Bosnia and Herzegovina (Laktaši, Banja Luka, Modriča and Višegrad). Adult H. contortus specimens were morphologically identified and confirmed using real-time quantitative PCR (rt-qPCR). Benzimidazole resistance was assessed by allele-specific rt-qPCR targeting the F200Y mutation in the β-tubulin isotype 1 gene. Statistically significant interspecies differences in β-tubulin genotype distribution were observed (p < 0.05), primarily driven by variation in the homozygous resistant (RR) genotype. High RR prevalence was detected in sheep (60%), lambs (50%) and roe deer (52.5%), whereas lower proportions were observed in chamois (27.3%) and goats (23.5%). Overall, 44.1% of all analyzed H. contortus isolates carried homozygous resistant alleles, indicating an advanced stage of benzimidazole resistance within this multi-host system. These findings demonstrate that benzimidazole resistance in H. contortus is not confined to domestic livestock but is also present in wild ruminants sharing the same grazing areas, consistent with circulation of resistant parasites within shared grazing systems.