Introduction With their remarkable flight capabilities, wild and captive birds play a pivotal role in the global dissemination of zoonotic pathogens including Chlamydia psittaci, 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. Methods 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 Chlamydia spp., and C. psittaci, 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. Results Chlamydia spp. was detected in 29.9% (95% CI: 25.2–34.9) of samples. Further, C. psittaci 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. Discussion The identification of C. psittaci 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.

The objective of the study was to assess the genetic diversity of the Tornjak shepherd dog by analyzing 10 micro- satellite loci. The dogs were divided into three main groups: Tornjaks from Bosnia and Herzegovina (TBA), Tornjaks from Croatia (THR), and a control group containing four subgroups: German Shepherd (GS), Belgian Shepherd (BS), crossbreeds of Tornjaks and other breeds (MIXT), and various other dog breeds (MIXA). The average number of genotypes (GN) and alleles (AN) was 19.2 and 8.4 (TBA), 8.2 and 5.4 (THR), 7.9 and 5.1 (BS), 4.4 and 3.6 (GS), 7.6 and 5.5 (MIXT), and 8.6 and 6.0 (MIXA), respectively. The average values of observed (HO) and expected heterozy- gosity (HE) were 0.7261 and 0.7392 (TBA); 0.7625 and 0.7139 (THR); 0.6857 and 0.6837 (BS); 0.4900 and 0.4640 (GS); 0.6786 and 0.6760 (MIXT); and 0.7067 and 0.7160 (MIXA), respectively. In the TBA population, “private” alleles were observed at all ten loci. The average overall inbreeding coefficient (F) value between the Tornjak and the control group was 0.0768. The AMOVA test revealed the highest degree of variation within the TBA group (55.43%), while no significant variations were observed in the control subgroup GS. The smallest differentiation for TBA was found with THR at 1.86% used the pairwise FST (pFST). The constructed Neighbor-Joining (NJ) dendrogram shows clear grouping of TBA and THR in comparison to the breeds of the control group. In relation to the data presented, a high level heterogeneity of has been established in the studied Tornjak population (TBA).

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.

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.

The etiology of transmissible viral proventriculitis (TVP) of broiler chickens has been discussed since its initial recognition 40 years ago. Regardless of its low direct impact on mortality rate, it leads to high economic losses in the broiler industry through reduction of food conversion, weakening of birds, and their increased susceptibility to pathogens. The aim of the present study was to examine the potential presence of TVP on the broiler chicken farms in Bosnia and Herzegovina, to characterize microscopic lesions, and to investigate the viruses implicated in etiology of TVP by PCR-based methods. In total, 143 diseased broiler chickens from 16 farms in Bosnia and Herzegovina were euthanized and subjected to necropsy and subsequent histopathology of proventriculi. A representative number of proventriculi samples (n = 50) that exhibited histopathologic changes were processed for molecular detection of chicken proventricular necrosis virus (CPNV), girovirus (GyV3), chicken anemia virus (CAV), and infectious bursal disease virus (IBDV) by PCR-based methods. In addition, samples of bursa of Fabricius (n = 39) and spleen (n = 50) were tested for IBDV. Histopathology revealed changes consistent with TVP in 39.8% (57/143) and LP (lymphocytic proventriculitis) in 2.1% (3/143) of samples. All 50 proventricular samples showed positivity to CPNV with Ct values ranging between 18 and 26. GyV3 was detected in eight samples (16%), with Ct values ranging from 11.1 to 27.5. The presence of CAV was more prominent (38%), with 19 positive broiler chickens (Ct ranging from 9.6 to 35.6). Pooled samples of spleen, bursa, and proventriculi from three farms were positive for IBDV. The obtained results represent the first documented data on TVP and the first record of CPNV and GyV3 presence in broiler farms from Bosnia and Herzegovina.

The current state of research on the anti‑SARS‑CoV‑2 potential of artemisinin‑related compounds has identified arteannuin B as a potent inhibitor of the nCoV‑2019BetaCov/Wuhan/WiV04/2019 and BetaCov/Italy/CDG1/2020 strains of the virus. The aim of this work was to fractionate the targeted sesquiterpenoid compounds, arteannuin B and artemisinin, from the complex matrix of the crude ethanolic leaf extract of Artemisia annua L. using high‑speed countercurrent chromatography (HSCCC) and to test the simplified or purified fractions against the genomically characterized Alpha SARS‑CoV‑2 variant in vitro. This is the first detailed in vitro anti‑SARS‑CoV‑2 study using an analytically characterized supercritical fluid extract of A. annua L. The preparative HSCCC method enabled the isolation of purified arteannuin B in a single chromatographic step, which was confirmed by LC‑ESI‑QTOF‑MS/MS. The MS data confirmed the selectivity of the HSCCC method for the targeted fractionation of artemisinin from the complex matrix, as it was successfully separated from the EtOH crude extract without co‑elution with arteannuin B. Antiviral activity determined by quantitative real‑time PCR (qRT‑PCR) yielded half‑maximal effective concentrations (EC₅₀) of 93.7 µg/mL (SC‑CO₂ extract), 173.5 µg/mL (EtOH extract), 187.3 µg/mL (artemisinin knockout fraction), 38.1 µg/mL (arteannuin B fraction), and >100 µg/mL (artemisinin). The arteannuin B fraction was highly active at 50 µg/mL (p < 0.0001) and 100 µg/mL (p < 0.0001), and inhibited the amplification of the SARS‑CoV‑2 N and RdRp genes by 84% and 100%, respectively. An important contribution of this study is the demonstration of the antiviral activity of arteannuin B against the Alpha variant of SARS‑CoV‑2, which is known to have increased infectivity and transmissibility.

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.

Simple Summary Ticks are ectoparasites with medical significance. They inhabit diverse environments and maintain close interactions with numerous vertebrate hosts. Ixodes ticks can transmit various pathogens to animals and humans. The aim here was to examine Ixodes ticks from Bosnia and Herzegovina to check for specific pathogens. This study found Rickettsia spp., Babesia spp., Anaplasma phagocytophilum, and Borrelia burgdorferi sensu lato in ticks from domestic animals. These findings highlight the need for the ongoing monitoring of ticks and tick-borne pathogens to protect animal and public health. Additionally, this study provides valuable insights into the occurrence and spread of these pathogens, emphasizing the importance of broader surveillance and control measures. Effective prevention, surveillance, and control of tick-borne diseases require urgent regional and international collaboration. Abstract Limited information is available regarding the presence of tick-borne pathogens and their distribution within Ixodes species in Bosnia and Herzegovina. This study aimed to identify Rickettsia spp., Babesia spp., Anaplasma phagocytophilum, and Borrelia burgdorferi sensu lato (s.l.) in Ixodes ticks collected from domestic and wild animals and vegetation in different regions across Bosnia and Herzegovina. A total of 7438 adult ticks, including 4526 Ixodes ricinus, Ixodes canisuga, and Ixodes hexagonus, were collected. Real-time PCR screening of 450 pooled I. ricinus samples revealed a 22.1% infection rate with at least one pathogen. Rickettsia spp. (6.3%) were found in ticks from dogs, cats, and goats, Babesia spp. (3.1%) in ticks from dogs and cattle, A. phagocytophilum (8.8%) in ticks from dogs, goats, and cattle, and B. burgdorferi s.l. (3.4%) in ticks from dogs and cats. Mixed infections with B. burgdorferi s.l. and A. phagocytophilum, as well as B. burgdorferi s.l. and Rickettsia spp., were found in two pools of I. ricinus from dogs and cats, respectively. Additionally, co-infection with Rickettsia spp. and A. phagocytophilum was confirmed in three tick pools from dogs and goats. Each tick from these pooled samples was individually retested to confirm the presence of pathogens. In the examined pooled samples of I. canisuga (1) and I. hexagonus (6), none of the tested pathogens were detected. Our findings represent the first detection of Rickettsia spp., Babesia spp., A. phagocytophilum, and B. burgdorferi s.l. in I. ricinus collected from domestic animals and vegetation in Bosnia and Herzegovina. Considering the established infection rates, the detection of tick-borne pathogens in adult ticks collected from domestic animals and vegetation enriches the current knowledge of the presence of tick-borne pathogens at the local, regional, national, and broader levels.

Dermacentor (D.) reticulatus ticks carry and transmit a wide range of pathogens to vertebrate hosts. Limited information is available about the existence of emerging tick-borne pathogens and the distribution of D. reticulatus in Bosnia and Herzegovina. The study aimed to investigate the occurrence and distribution of D. reticulatus and to detect the presence of Anaplasma spp., Borrelia (B.) burgdorferi s.l., Rickettsia spp., and Babesia spp. in samples originating from questing ticks and ticks collected from domestic animals in various regions of Bosnia and Herzegovina. A total of 402 collected D. reticulatus ticks were widely distributed throughout the country. Of the 41 pools consisting of 205 individual D. reticulatus ticks, 21 (51.2%) indicated the presence of Rickettsia spp., 17 (41.4%) of Babesia spp., 2 (4.8%) of Anaplasma spp., and 1 (2.4%) of B. burgdorferi s.l. after real-time PCR screening. Our study indicates that D. reticulatus has significantly expanded its distribution and host range in Bosnia and Herzegovina. Moreover, our results represent the first detection of Babesia spp. in D. reticulatus in Bosnia and Herzegovina. Given the demonstrated presence of emerging pathogens in questing and feeding ticks, there is an urge to establish a surveillance system for ticks and tick-borne pathogens in Bosnia and Herzegovina.