Objective: The aim of this study was to examine the solvent conditions suitable for preparing samples of total mixed ration (TMR) silage and ruminal fluid, with or without chitosan inclusion, to achieve more sensitive, specific, and repeatable analyses. Material and Methods: The metabolome contained in the sample was determined using a ultrahigh-performance liquid chromatography-high-resolution mass spectrometry quadrupole using samples in the form of TMR silage (silage with 0.5% inclusion of chitosan, silage without chitosan addition) and rumen fluid (rumen fluid containing 0.5% chitosan, ruminal fluid without chitosan) with and without the addition of chitosan, as well as three different solvent ratios: 50% water MS grade: 50% methanol (MeOH) MS grade (1), 20% water MS grade: 80% MeOH MS grade (2), and 0% water MS grade: 100% MeOH MS grade (3). Results: The TMR silage samples contained 311 metabolites, amino acid compounds predominating, followed by fatty acids and amines. Fatty acids, organic phosphate, and amines dominated the 39 metabolites found in rumen fluid samples. In the TMR silages, 100% MeOH seemed to be able to distinguish samples more effectively than other solvents; moreover, cinnamic acid, D-(+)-proline, and L(+)-ornithine were the three highest variable importance for projection values among prospective metabolite markers for silage samples. Whereas the use of a 50% MeOH to 50% water solvent ratio achieved the best discrimination results in rumen fluid samples, the three highest variable importance for projection values among prospective metabolite markers for ruminal fluid samples were triethyl phosphate, dibenzylamine, and phosphoric acid. Conclusion: 100% MeOH is able to distinguish TMR silage, while 50% MeOH to 50% water is best for ruminal fluid samples. J. Adv. Vet. Anim. Res., 12(2): 589–600, June 2025 http://doi.org/10.5455/javar.2025.l923

Objective: The aim of this study was to examine the solvent conditions suitable for preparing samples of total mixed ration (TMR) silage and ruminal fluid, with or without chitosan inclusion, to achieve more sensitive, specific, and repeatable analyses. Material and Methods: The metabolome contained in the sample was determined using an ultra-high-performance liquid chromatography-high-resolution mass spectrometry quadrupole using samples in the form of TMR silage (silage with 0.5% inclusion of chitosan, silage without chitosan addition) and rumen fluid (rumen fluid containing 0.5% chitosan, ruminal fluid without chitosan) with and without the addition of chitosan, as well as three different solvent ratios: 50% water MS grade: 50% methanol (MeOH) MS grade (1), 20% water MS grade: 80% MeOH MS grade (2), and 0% water MS grade: 100% MeOH MS grade (3). Results: The TMR silage samples contained 311 metabolites, amino acid compounds predominating, followed by fatty acids and amines. Fatty acids, organic phosphate, and amines dominated the 39 metabolites found in rumen fluid samples. In the TMR silages, 100% MeOH seemed to be able to distinguish samples more effectively than other solvents; moreover, cinnamic acid, D-(+)-proline, and L(+)-ornithine were the three highest variable importance for projection values among prospective metabolite markers for silage samples. Whereas the use of a 50% MeOH to 50% water solvent ratio achieved the best discrimination results in rumen fluid samples, the three highest variable importance for projection values among prospective metabolite markers for ruminal fluid samples were triethyl phosphate, dibenzylamine, and phosphoric acid. Conclusion: 100% MeOH is able to distinguish TMR silage, while 50% MeOH to 50% water is best for ruminal fluid samples. [J Adv Vet Anim Res 2025; 12(2.000): 589-600]

Objective: To describe the clinical presentation, diagnostic imaging findings, and management of a rare case of bilateral temporomandibular joint (TMJ) ankylosis combined with dysplasia in a cat. Materials and Methods: A 4-month-old Bombay male cat, normally immunized and dewormed. Radiographic studies: Details of imaging techniques used to diagnose TMJ ankylosis and dysplasia: computed tomography (CT) scans were used to visualize the joint structures in greater detail using CT. Results: Out of the 21 blood biochemical abnormalities found in the laboratory test, creatinine, albumin, albumin/globulin, and total cholesterol all showed a significant decrease. The levels of phosphorus and α-amylase were marginally elevated. A three-dimensional CT scan revealed a malformed fusion of the right TMJ zygomatic arch and bilateral mandibular coronal process. Both TMJ dysplasia and ankylosis were discovered by the biochemical and physical 3-dimensional CT scans. Conclusion: The findings underscore the importance of a thorough clinical examination and imaging studies to assess the extent of the ankylosis and any associated dysplastic changes. This case emphasizes the need for increased awareness of TMJ disorders in felines and encourages further research into effective treatment protocols. J. Adv. Vet. Anim. Res., 12(2): 321–326, June 2025 http://doi.org/10.5455/javar.2025.l899 

Objective: To describe the clinical presentation, diagnostic imaging findings, and management of a rare case of bilateral temporomandibular joint (TMJ) ankylosis combined with dysplasia in a cat. Materials and Methods: A 4-month-old Bombay male cat, normally immunized and dewormed. Radiographic studies: Details of imaging techniques used to diagnose TMJ ankylosis and dysplasia: computed tomography (CT) scans were used to visualize the joint structures in greater detail using CT. Results: Out of the 21 blood biochemical abnormalities found in the laboratory test, creatinine, albumin, albumin/globulin, and total cholesterol all showed a significant decrease. The levels of phosphorus and α-amylase were marginally elevated. A three-dimensional CT scan revealed a malformed fusion of the right TMJ zygomatic arch and bilateral mandibular coronal process. Both TMJ dysplasia and ankylosis were discovered by the biochemical and physical 3-dimensional CT scans. Conclusion: The findings underscore the importance of a thorough clinical examination and imaging studies to assess the extent of the ankylosis and any associated dysplastic changes. This case emphasizes the need for increased awareness of TMJ disorders in felines and encourages further research into effective treatment protocols. [J Adv Vet Anim Res 2025; 12(2.000): 321-326]

African swine fever (ASF) is a highly fatal viral haemorrhagic disease affecting pigs. This study uses official surveillance data to investigate the persistence and spread of ASF in wild boar populations in Serbia from 2020 to 2024. A total of 480 ASF cases were analysed, with spatiotemporal clustering conducted via SaTScan™ and proximity analyses performed in the Quantum Geographic Information System. The yearly prevalence of ASF in wild boar rose steadily from 0.66% in 2020 to 1.47% in 2023. Seasonal trends showed a significant increase in cases during winter (68%) and spring (24%). Five distinct clusters were identified, predominantly near the borders with North Macedonia and Bulgaria, suggesting potential cross-border transmission. Proximity to major roads was significantly associated with ASF case distribution (p  0.01), while proximity to water bodies and elevation showed no consistent pattern. The findings highlight critical gaps in current passive surveillance systems, which likely underestimate ASF prevalence. The study underscores the need for enhanced surveillance in remote and wooded areas and proposes strategies to improve ASF monitoring and control in wild boar populations.Contribution: This study highlights the feasibility of cost-effective, non-invasive surveillance methods for ASF detection, offering critical insights for low-income countries where resources for intensive disease monitoring are limited. By demonstrating how environmental and anthropogenic factors drive ASF dynamics, this research provides actionable strategies for improving regional and global ASF control efforts.

African swine fever (ASF) is a highly fatal viral haemorrhagic disease affecting pigs. This study uses official surveillance data to investigate the persistence and spread of ASF in wild boar populations in Serbia from 2020 to 2024. A total of 480 ASF cases were analysed, with spatiotemporal clustering conducted via SaTScan™ and proximity analyses performed in the Quantum Geographic Information System. The yearly prevalence of ASF in wild boar rose steadily from 0.66% in 2020 to 1.47% in 2023. Seasonal trends showed a significant increase in cases during winter (68%) and spring (24%). Five distinct clusters were identified, predominantly near the borders with North Macedonia and Bulgaria, suggesting potential cross-border transmission. Proximity to major roads was significantly associated with ASF case distribution (p  0.01), while proximity to water bodies and elevation showed no consistent pattern. The findings highlight critical gaps in current passive surveillance systems, which likely underestimate ASF prevalence. The study underscores the need for enhanced surveillance in remote and wooded areas and proposes strategies to improve ASF monitoring and control in wild boar populations. Contribution: This study highlights the feasibility of cost-effective, non-invasive surveillance methods for ASF detection, offering critical insights for low-income countries where resources for intensive disease monitoring are limited. By demonstrating how environmental and anthropogenic factors drive ASF dynamics, this research provides actionable strategies for improving regional and global ASF control efforts.

The widespread use of parabens has led to their accumulation in aquatic environments. This study examined the effects of methylparaben on rainbow trout, dividing 96 fish into control and treatment groups (1 mg/L, 5 mg/L, and 8 mg/L). Results showed dose-dependent weight loss, altered hepatosomatic indices, increased serum urea, uric acid, and Malondialdehyde (MDA) levels, and decreased Glutathione Peroxidase (GSH-Px) activity. Histopathological analysis revealed liver and kidney abnormalities in treated groups, including hepatocyte degeneration, proliferation in the bile duct, glomerular atrophy, reduced haematopoietic tissue, increased melanomacrophage centres, necrosis and fibrosis.Contribution: These findings highlight methylparaben’s toxic effects, emphasising the need for stricter regulations and further research to safeguard aquatic ecosystems and understand its impact on aquatic organisms.

The widespread use of parabens has led to their accumulation in aquatic environments. This study examined the effects of methylparaben on rainbow trout, dividing 96 fish into control and treatment groups (1 mg/L, 5 mg/L, and 8 mg/L). Results showed dose-dependent weight loss, altered hepatosomatic indices, increased serum urea, uric acid, and Malondialdehyde (MDA) levels, and decreased Glutathione Peroxidase (GSH-Px) activity. Histopathological analysis revealed liver and kidney abnormalities in treated groups, including hepatocyte degeneration, proliferation in the bile duct, glomerular atrophy, reduced haematopoietic tissue, increased melanomacrophage centres, necrosis and fibrosis. Contribution: These findings highlight methylparaben’s toxic effects, emphasising the need for stricter regulations and further research to safeguard aquatic ecosystems and understand its impact on aquatic organisms.