Adis Softić

Animal disease outbreaks at the wildlife-livestock interface threaten animal health, ecosystems, food systems, and, for zoonoses, human health, while veterinary authorities must maintain preparedness with limited resources. However, existing preparedness tools characterize capacity, exposure, or risk largely in isolation. This study developed a framework to assess whether preparedness is commensurate with disease-specific challenge. Questionnaire-derived Preparedness capacity for 26 European countries was estimated using Item Response Theory and adjusted for socioeconomic and governance context. Context-adjusted preparedness was then compared with disease-specific Challenge Indices combining livestock population, livestock density, animal imports, and wildlife reservoir abundance, for two diseases at the wildlife-livestock interface: avian influenza (AI) in domestic poultry and African swine fever (ASF) in domestic pigs. Countries were classified into four Preparedness-Challenge profiles and summarized regionally, with uncertainty propagated by Monte Carlo simulation and decomposed by variance-based sensitivity analysis. The Underprepared (high challenge) profile included 9 countries for AI and 10 for ASF and was the most common profile in Southern and Western Europe for both diseases, and in Central Europe for ASF. Livestock weighting sharpened this pattern, with this profile holding 66-91% of poultry and 79-97% of pigs in Central, Southern, and Western Europe. In Northern Europe, 76% of pigs were in the Prepared (high challenge) profile. Preparedness estimation was the largest source of uncertainty in country-level classifications and regional AI results, whereas Challenge Index weighting dominated regional ASF results. Countries with less preparedness relative to their challenge tended to have greater recorded outbreak burden (concordance probability: AI 0.63-0.70 for continuous burden measures, ASF 0.58-0.85 for all measures). Preparedness gaps were therefore disease-specific, and unweighted country summaries obscured where livestock exposure was concentrated. Disease-specific, livestock-weighted comparisons offer veterinary authorities a transparent basis for prioritizing preparedness investment, complementing established One Health tools.

Background/Objectives: Establishing drowning as the cause of death remains one of the most challenging tasks in forensic pathology, particularly when classical autopsy findings are absent, nonspecific, or compromised by decomposition. Molecular biomarkers may provide objective adjunctive evidence in such cases. This study investigated the diagnostic utility of Aquaporin-5 (AQP5) and Surfactant Protein A2 (SP-A2) measured in renal tissue as potential biomarkers of freshwater drowning and evaluated the influence of different freshwater environments on their expression. Methods: A controlled experimental study was performed using 29 adult Wistar rats allocated to freshwater drowning groups (Boračko Lake, n = 11; Jablaničko Lake, n = 11) and non-drowning control groups subjected to mechanical asphyxia followed by one hour of postmortem immersion (n = 7). Kidney tissues were collected within a maximum of 5 min after death and analyzed for AQP5 and SP-A2 concentrations using enzyme-linked immunosorbent assay (ELISA). Group differences were evaluated using Kruskal–Wallis and Dunn’s post hoc tests, while effect sizes were estimated using Cliff’s delta. Receiver operating characteristic (ROC) curve analysis was performed to assess the diagnostic performance of each biomarker in distinguishing drowning from non-drowning deaths. Results: AQP5 concentrations differed significantly among groups (χ² = 16.05, p = 0.00033), with large effect sizes observed in all pairwise comparisons. However, diagnostic performance was poor, as ROC analysis demonstrated no significant discrimination between drowning and non-drowning cases (AUC = 0.534, 95% CI: 0.314–0.754; p = 0.762). In contrast, SP-A2 showed significant intergroup differences (χ² = 6.69, p = 0.035) and moderate diagnostic accuracy for drowning identification (AUC = 0.752, 95% CI: 0.567–0.937; p = 0.008), with a sensitivity of 77.2% and specificity of 81.8% at the optimal threshold. Notably, AQP5 expression appeared suggestive of environmental modulation, whereas SP-A2 demonstrated greater relevance to the drowning process itself. Conclusions: Renal SP-A2 showed moderate diagnostic utility as a supportive biomarker for drowning, whereas AQP5 lacked sufficient discriminatory power for cause-of-death determination. The marked variability of AQP5 across freshwater environments suggests that it reflects environmental modulation rather than drowning-specific biological responses. Although neither marker is suitable as a standalone diagnostic tool, renal SP-A2 may represent a useful adjunct in forensic drowning investigations, particularly when pulmonary tissue is unavailable or unsuitable for analysis. These findings should be considered preliminary and applicable only to freshwater drowning conditions, as saltwater drowning was not investigated in the present study. Further validation in human forensic casework and larger experimental cohorts is warranted.

Bats are one of the most widespread, but also the least researched groups of mammals. Their guano is an important source of DNA because it is often readily available even if bats are not, and it may be used to reliably identify different species of bats. According to the most recent data, there are 31 species of bats recorded in Bosnia and Herzegovina, identified by the morphometric and acoustic methods. This study had the goal of studying bat diversity in Bosnia and Herzegovina by sequencing the COI gene from guano samples and comparing the results with the results of morphometric measurements. A total of 10 bat species were identified through morphometric analysis, while 13 bat species were identified through the sequencing of COI gene from guano samples, including a new species for the country: Rhinolophus mehelyi. Sequencing has increased the number of species recorded on each locality and resolved inconclusive morphological findings except in the case of M. myotis/blythii. The results indicate that the biodiversity of bats in Bosnia and Herzegovina is larger than estimated and that COI sequencing from guano can be used for regular monitoring and for identification of sites of special importance to bats.

Antimicrobial resistance (AMR) in poultry-associated Escherichia coli (E. coli) is a persistent One Health concern, particularly when ESBL/pAmpC determinants co-occur with resistance to multiple antimicrobial classes. Between March and October 2024, we investigated commensal E. coli from three interconnected compartments of the poultry production chain in Bosnia and Herzegovina (parent-breeder flocks, commercial broiler farms, hatchery-associated material). A total of 333 samples were examined, and 99 E. coli isolates were recovered (29.7%). Phenotypic characterization included ESBL confirmation, disk diffusion susceptibility testing, and EUVSEC broth microdilution. Targeted real-time PCR assays were used to screen key ESBL/pAmpC-associated genes and selected carbapenemase and plasmid-mediated colistin resistance targets within the targeted panel. ESBL phenotypes were detected in 52/99 isolates (52.5%), and multidrug resistance was highly prevalent across compartments (93/99; 93.9%). ESBL/pAmpC-associated genes were detected in 91/99 isolates (91.9%), with bla_TEM predominating. Gene pattern analysis indicated that bla_TEM occurred most frequently as a single determinant and as part of the predominant multi-gene combinations, most notably bla_TEM + bla_CMY and bla_TEM + bla_CTX-M, while bla_SHV was sporadic. Carbapenemase genes (bla_KPC, bla_NDM, bla_GES, bla_OXA-48) and mcr-1 to mcr-9 were not detected. Overall, our findings indicate a substantial ESBL/MDR burden throughout the poultry production chain, supporting the need for strengthening antimicrobial stewardship and biosecurity measures across both farms and hatcheries.

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.

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.

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.

Arthropods are a highly diverse group of invertebrates, found in almost every habitat and constituting more than 80% of all recognized animal species. Their diversity is evident in the numerous roles they fulfill in ecosystems, including crucial functions like pollination, decomposing organic matter, and regulating pest populations. Despite their importance in maintaining ecological balance, certain arthropods can become pests in agriculture and storage, leading to crop losses and contamination of stored products. Additionally, some arthropods, such as mosquitoes, ticks, and fleas, transmit microorganisms such as bacteria, viruses, and parasites, resulting in diseases in humans and animals. Their role as vectors of pathogens is a vital research area in veterinary medicine, focusing on understanding disease transmission mechanisms and developing control strategies. Therefore, this chapter emphasizes the importance of arthropod diversity in the veterinary context and highlights various arthropod species and the pathogens they transmit.

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.