Chlamydia abortus is an obligate intracellular Gram-negative pathogenic bacterium widely distributed globally. This pathogen primarily infects humans and various domestic animals, causing abortions in pregnant animals. It poses a significant threat to public health and the animal husbandry industry, leading to substantial economic losses. In this study, specific primers and a Proofman fluorescent probe for LMTIA (Ladder Melting Temperature Isothermal Amplification) were designed based on the genome of the C. abortus S26/3 strain through sequence alignment and screening of specific target sequences. These primers and probes were used to develop a rapid Proofman-LMTIA assay for detecting C. abortus. The reaction system and temperature were optimized, and the specificity, sensitivity, and reproducibility of the assay were evaluated. The optimal amplification temperature for Proofman-LMTIA was 59.5 degrees C, and the reaction time was less than 30 minutes. The established assay had a minimum detection limit of 101 fg/mu L per test, indicating a high analytical sensitivity. The assay also showed excellent stability, with coefficients of variation below 3% in inter-group and intra-group reproducibility tests. In summary, the rapid and effective Proofman-LMTIA method for detecting C. abortus developed in this study provides a novel technical tool for the clinical diagnosis and control of C. abortus infections.
This study included dairy farms registered as milk producers in the entity of Republika Srpska, Bosnia and Herzegovina. A total of 180 milk samples originating from cows were collected from dairy farms across several locations during 2024 and 2025. Out of the 180 tested raw milk samples obtained from dairy cows suspected to suffer from clinical or subclinical mastitis the presence of Staphylococcus aureus was confirmed in 44 samples, representing 24.4% (SD: 3.2%; 95% CI: 18.2% - 30.7%) of all analyzed samples. The presence of one or more se genes was detected in nine isolates (20.5%; 95% CI: 11.0% - 33.7%). Among the total of 44 S. aureus isolates, only one isolate exhibited phenotypic resistance to cefoxitin using ETE ST (R) Cefoxitin, which was molecularly confirmed by detection of the mecA gene. All analyzed S. aureus isolates showed resistance to at least one of the tested antimicrobial agents, while 25% (95% CI: 12.2% - 37.8%) of isolates showed resistance to two antimicrobial agents. A total of 15.9% (95% CI: 6.6% - 30.1%) of isolates displayed antimicrobial multidrug resistance (MDR ), defined as resistance to three or more classes of antimicrobial drugs.
Ascaridia compar (Schrank, 1790) is a nematode parasite of wild galliform birds that has long been surrounded by taxonomic uncertainty due to overlapping morphological features with several historically described species. In this study, we provide molecular evidence supporting the species identity and preliminary phylogenetic placement of A. compar isolated from western capercaillie (Tetrao urogallus) in Bosnia and Herzegovina. Morphological analysis of seven adult nematodes (six females, one male) revealed diagnostic traits consistent with assignment to the genus Ascaridia, however, because morphological characters overlap among several historically described grouseassociated taxa, species-level identification was subsequently supported by molecular analysis of the ITS1, cox1, and cox3 gene regions and comparison with homologous sequences available in GenBank. Phylogenetic analysis supported the identification of the capercaillie-derived specimens as A. compar, with high bootstrap support distinguishing them from A. galli, A. columbae, and A. nymphii. Pairwise distance analysis showed a 1-4% divergence from the Italian A. compar isolate, and significantly greater divergence from congeneric species. This study contributes preliminary molecular evidence relevant to a longstanding taxonomic ambiguity and supports the distinction of A. compar from currently compared congeners, although broader sampling across multiple hosts and populations is needed to confirm these patterns. The presented findings highlight the relevance of integrating morphological and molecular data in nematode systematics and point to the need for future studies on parasite dynamics in declining grouse populations.
The study aimed to assess the potential to reduce antibiotic usage during the dry period of dairy cows and the associated environmental impacts by simulating potential cephapirin (Cefa-Safe) application according to the recording season and udder health status of the dairy cow population in Croatia. Official milk recording data from the Croatian Agency for Agriculture and Food (HAPIH) collected between 2013 and 2022 were used for the analysis. Based on these data, cows were classified according to udder health status: healthy (<200000 cells/mL), subclinical mastitis (200000-400000 cells/mL), and clinical mastitis (>400000 cells/mL). Using these proportions, the total amount of cephapirin, its economic value, and potential environmental emission under the standard blanket dry cow therapy (BDCT) scenario were simulated. The analysis was conducted by season (spring, summer, autumn, winter). The highest estimated antibiotic usage and emissions were observed during summer in Holstein cows and in spring in Simmental cows, whereas the lowest values were recorded in winter for both breeds. The calculated risk quotients (RQ = PEC/PNEC) exceeded the threshold value RQ > 1 in all seasons, indicating a persistent environmental risk. The economic estimations followed the same pattern, with the highest costs recorded during the warmer part of the year. The results confirm that seasonal variation affects udder health status and consequently the amount of antibiotics potentially required during the dry period. The proposed model enables a quantitative assessment of the potential for reducing antibiotic usage and provides a scientific basis for transitioning from standard to selective dry cow therapy (SDCT) within the framework of sustainable dairy production.
Laparoscopic cholecystectomy in dogs induces hemodynamic and respiratory changes due to pneumoperitoneum, thereby making fluid therapy a central element of perioperative management. This retrospective, single-center study evaluated the effect of two crystalloid strategies: restrictive (5 mL/kg/h) and liberal (10 mL/kg/h), in 104 dogs anesthetized between 2014 and 2025 and subjected to laparoscopic cholecystectomy. The primary outcome was perioperative mortality (<= 7 days). Secondary outcomes included cardiovascular instability, hypothermia, vasopressor requirement, transfusion requirement, acute kidney injury, hospital resource utilization and short-term recovery. Analyses included between-group comparisons, multivariate logistic regression, and propensity to score matching (1:1). Mortality was low and similar between strategies. The liberal strategy was associated with higher frequency of hypothermia and lower minimum intraoperative temperature, while the restrictive strategy required higher maximum vasopressor doses, without increased severe events. In multivariate models, longer anesthetic time was associated with cardiovascular instability and hypothermia; higher body weight was protective against hypothermia. Findings remained consistent after matching. Therefore, in canine laparoscopic cholecystectomy, restrictive (5 mL/kg/h) and liberal (10 mL/kg/h) strategies showed similar clinical results; choice should be individualized based on each patient's clinical status.
