Veterinary Medicine International最新文献

{"title":"Polyomavirus Infection in Two Young Parrots: Assessment of Viral Distribution in Organs and Affected Tissues.","authors":"Gaia Casalino, Davide Messina, Clara Tolini, Giancarlo Bozzo, Antonio Lavazza, Francesco D'Amico, Antonella Bove, Francesco Pellegrini, Francesco D'Onghia, Diana Romito, Vito Martella, Canio Buonavoglia, Antonio Camarda, Elena Circella","doi":"10.1155/vmi/2135685","DOIUrl":"10.1155/vmi/2135685","url":null,"abstract":"<p><p>Avian polyomavirus (APyV) is responsible for diseases in birds belonging to several avian orders, including Psittaciformes, Anseriformes, and other susceptible taxa. APyVs appear to be able to replicate in various organs of susceptible bird species. This study aimed to investigate (i) the distribution of APyV in the organs of two parrot chicks belonging to different species, Moluccan eclectus (<i>Eclectus roratus</i>) and blue-winged macaw (<i>Primolius maracana</i>), which died due to APyV infection and (ii) the lesions associated with the infection. By negative-staining electron microscopy, well-defined viral particles morphologically compatible with polyomavirus and polyomavirus-like particles were identified in organs. Detection of APyV was confirmed by real-time PCR, and BLAST analysis of the complete genome revealed 99.96% identity with a budgerigar fledging disease polyomavirus strain. The amount of DNA copy number/ng of DNA ranged from 2.47 × 10³ found in the myocardium to 3.90 × 10⁶ found in the feather quills of the eclectus and from 1.07 × 10¹ to 1.82 × 10⁷ detected in the brain and intestine of the blue-winged macaw, respectively. According to the findings of inclusion bodies, viral particles, and DNA, the lung appeared as one of the most severely affected tissues. Even if inclusion bodies were not detected, severe lesions were found in the myocardium, bursa of Fabricius, and spleen. The results of this study highlight the ability of APyV to spread widely, leading to severe lesions in the organs of infected birds. The most significant amount of viral DNA detected in the gut and feathers highlights the relevance of feces and feathers as useful samples for the in vivo diagnosis.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"2135685"},"PeriodicalIF":2.3,"publicationDate":"2026-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13500891/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148813868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Histopathological Features and Clinical Findings of Feline Chronic Gingivostomatitis (FCGS) in Cats (<i>Felis catus</i>) at Gloriavet Clinic, Bandung.","authors":"Febriani Rohmatul Azizah, Grace Julianty Hutomo, Chrysanti Murad, Shafia Khairani","doi":"10.1155/vmi/2438106","DOIUrl":"10.1155/vmi/2438106","url":null,"abstract":"<p><p>Feline chronic gingivostomatitis (FCGS) is a chronic inflammatory condition of the oral cavity characterized by distinctive lesions involving various areas of the mucosa. Histopathological examination was performed to differentiate FCGS from other inflammatory conditions and to assess its severity for diagnostic purposes. The aim of this study was to evaluate the clinical findings and histopathological features of oral lesions in cats diagnosed with FCGS and to examine their association with patient-related factors, including breed, sex, age, and housing conditions. A semiquantitative approach was used to assess clinical findings using the Stomatitis Disease Activity Index (SDAI) and histopathological lesion scoring. The results revealed pathological findings such as erythema, swelling, hemorrhage, and ulceration, with the most severe lesions commonly observed in the gingival region associated with periodontal problems. Histopathological findings included epithelial hyperplasia, intraepithelial inflammation cell infiltration, spongiosis, and ulceration as indicators of epithelial damage, along with inflammatory cell infiltration within the lamina propria and submucosa, with varying degrees of severity. Statistical analysis showed that patient-related variables had no significant influence on the severity of either clinical or histopathological findings (<i>p</i> > 0.05). These findings suggest that the progression and severity of FCGS are closely associated with immune responses to multiple pathogens within the oral mucosa rather than by individual patient factors.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"2438106"},"PeriodicalIF":2.3,"publicationDate":"2026-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13494511/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Virulence of <i>Mycobacterium avium</i> Subsp. <i>hominissuis</i> Isolated From Feline Thyroid and Pancreatic Tissues in a Murine Infection Model.","authors":"Adeliane Castro da Costa, Fábio Muniz de Oliveira, Fabrício Camargo, Lázaro Moreira Marques-Neto, Ísis Assis Braga, Danieli Brolo Martins, André Kipnis, Ana Paula Junqueira-Kipnis","doi":"10.1155/vmi/7207350","DOIUrl":"10.1155/vmi/7207350","url":null,"abstract":"<p><p>Mycobacteria infection is a serious public health problem and has been described as causing diseases in domestic animals. The objective of this study was to evaluate the immunopathogenesis of <i>Mycobacterium avium</i> subspecies <i>hominissuis</i> isolated from a domestic cat in mice. Mice were intravenously infected with 10⁶ CFU, and the spleen, lung, liver, and submandibular glands were collected for a histopathology study and bacillary load evaluation. Lung cells, such as macrophages, NK1.1 cells, and CD4⁺ and CD8⁺ T lymphocytes were evaluated by flow cytometry. The infection spread to the liver, lungs, lymph nodes, and submandibular salivary gland, causing severe necrotic lesions and inducing increased levels of lung macrophages, NK, and dendritic cells, as well as specific CD4⁺IFN-γ⁺ and CD8⁺IFN-γ⁺ T lymphocytes. Thus, <i>Mycobacterium avium</i> subsp<i>. hominissuis</i> presented here induced pathogenicity and immune responses in the mouse model, as other mycobacterial infections, indicating that this bacterium could induce serious disease in animals.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"7207350"},"PeriodicalIF":2.3,"publicationDate":"2026-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13494856/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Splenic Lesions in Dogs: A Retrospective Epidemiological and Pathological Study.","authors":"Hathali de Las Mercedes Sanchez Mata, Janeth Agudelo Quintero, Diego Alfonso Aranzazu Taborda, David Alzate Velásquez, Julián David Muñoz Duque","doi":"10.1155/vmi/9086933","DOIUrl":"10.1155/vmi/9086933","url":null,"abstract":"<p><p>Splenic neoplastic and non-neoplastic lesions are frequently diagnosed in dogs and often exhibit overlapping clinical signs and gross morphological features, rendering histopathological examination essential for definitive diagnosis, therapeutic planning, and prognostic assessment. In Colombia, epidemiological data regarding these lesions remain limited, whereas international studies report considerable heterogeneity in the relative frequencies of neoplastic and non-neoplastic conditions. This study aimed to characterize the histopathological spectrum of splenic lesions in dogs and provide updated regional epidemiological data. A retrospective analysis was conducted on splenic specimens obtained from dogs subjected to total splenectomy between January 2019 and June 2024. All samples were processed and examined histopathologically at three veterinary pathology laboratories in Medellín, Colombia. Descriptive statistical analyses were performed considering year of submission, patient demographic data, morphological diagnosis, and concurrent histopathological findings. A total of 424 cases were evaluated, of which 282 (66.5%) corresponded to non-neoplastic lesions and 142 (33.5%) to neoplastic processes. The mean age of dogs with non-neoplastic lesions was 9.5 (< 1-16) years, whereas dogs with neoplastic lesions had a mean age of 10.2 (4-17) years. Splenitis was the most frequent non-neoplastic diagnosis, accounting for 54.6% of cases and demonstrating a higher prevalence than reported in prior literature. Among malignant splenic neoplasms, hemangiosarcoma was the predominant diagnosis, representing 69.2% of cases. This study demonstrates a high prevalence of non-neoplastic splenic lesions, particularly splenitis, and a frequent occurrence of multiple concurrent morphological diagnoses in non-neoplastic cases. These findings provide novel regional epidemiological data and contribute original information to international literature, underscoring the importance of histopathological splenic evaluation for accurate diagnosis and clinical decision-making.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"9086933"},"PeriodicalIF":2.3,"publicationDate":"2026-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13498858/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phytochemical Supplementation in Grazing Dairy Cows: Effects on Milk Production, Metabolic Status, Hematology, and Postpartum Ovarian Activity.","authors":"Manuela Comesaña, Ana Meikle, Diego Mattiauda, Marisela Arturo-Schaan, Aline C Dall-Orsoletta, Rodrigo Vivián, Gretel Ruprechter, María de Lourdes Adrien","doi":"10.1155/vmi/2399561","DOIUrl":"10.1155/vmi/2399561","url":null,"abstract":"<p><p>This study evaluated the effects of three treatments in dairy cows under pasture-based systems during the transition and early lactation periods: one group supplemented with a phytochemical mix (polyphenols, curcuminoids, piperine, and trans-cinnamaldehyde, PHY), another group supplemented with monensin (MON), and a third group with no additive, used as the control (CTL). Sixty Holstein cows (primiparous and multiparous) were assigned to these treatments from 30 days prepartum to 60 days postpartum. Productive, metabolic, endocrine, hematological, and reproductive parameters were analyzed. Results showed that both PHY and MON increased milk yield in primiparous cows compared to CTL, whereas no treatment differences were observed in multiparous cows. MON supplementation was associated with higher serum cholesterol concentrations, particularly in primiparous cows. Total leukocyte counts did not differ between treatments; however, a treatment effect was observed for monocytes in multiparous cows. Energy balance indicators (nonesterified fatty acids, beta-hydroxybutyrate, and glucose) and calcium and phosphorus were not significantly influenced by treatments and remained within physiological ranges. In conclusion, both MON and the phytochemical mix supported adaptation during the transition period, although their effects differed by parity and physiological response. MON was more closely associated with lipid metabolism, while the phytochemical mix may have contributed to subtle immune-related adjustments, warranting further investigation.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"2399561"},"PeriodicalIF":2.3,"publicationDate":"2026-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13494660/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798313","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of Gastrointestinal Parasites in Grazing Sheep in Mountain, Piedmont, and Savanna Conditions of Casanare, Colombia.","authors":"José Alfredo Cáceres-Díaz, Ronaldo Tarache-Carreño, Daniela Caro-Estrada, Gloria Estefanía Pastrana-Aguirre, Ciro Ortiz-Valdes, Luis Edgar Tarazona-Manrique","doi":"10.1155/vmi/1404541","DOIUrl":"10.1155/vmi/1404541","url":null,"abstract":"<p><p>Gastrointestinal parasitosis represents a significant constraint on sheep productivity in Colombian tropical production systems; however, their regional epidemiological dynamics remain insufficiently characterized. Therefore, the objective of this study was to characterize the prevalence, associations, coinfestations, and health-related correlations of gastrointestinal parasites in grazing sheep from the Mountain, Piedmont, and Savanna ecoregions of the Department of Casanare, Colombia. A cross-sectional observational study was conducted, including quantitative and qualitative coprological examinations, assessments of environmental and production-related factors, and animal-level clinical indicators (FAMACHA score and body condition score). Data were analyzed using bivariate analyses (chi-square or Fisher's exact test) and nonparametric comparative tests (Mann-Whitney <i>U</i> or Kruskal-Wallis test), as appropriate. The overall prevalence was high (78.3%), with higher values in the Mountain ecoregion (90%) compared with the Piedmont (76.7%) and the Savanna (76.1%). A statistically significant association (<i>p</i> < 0.05) was identified between municipality, the water source, and parasitic prevalence. Parasitic burden differed significantly among ecoregions (<i>p</i> < 0.05). No correlation was observed between animal-level clinical indicators and parasite burden (<i>ρ</i> < 0.065). The distribution of parasite load was aggregated, with a predominance of mild infestations and a small subset of animals harboring burdens exceeding 5000 eggs per gram. Frequent coinfestations among nematodes, protozoa, and cestodes were identified, involving low-intensity associations. Despite the high prevalence, no clinically relevant health alterations were detected. Gastrointestinal parasitosis exhibited high prevalence, an aggregated distribution of parasite burden, and frequent low-intensity coinfestations, with variation among ecoregions and no association with clinical indicators. These findings suggest a dynamic process modulated by environmental and management-related factors, indicating host resilience and highlighting the need for targeted control strategies and longitudinal monitoring.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"1404541"},"PeriodicalIF":2.3,"publicationDate":"2026-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13494515/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798442","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular Insights Into the Phylogenetic Diversity, Virulence Determinants, and Antimicrobial Resistance Pattern of <i>Escherichia coli</i> From Milk of Mastitis-Affected Cows.","authors":"Md Mominul Islam, Ayesha Akter, Md Sadequl Islam, Mahfuzul Islam, S M Harun Ur Rashid, Mirza Mienur Meher, Jahagir Alam","doi":"10.1155/vmi/3499907","DOIUrl":"10.1155/vmi/3499907","url":null,"abstract":"<p><p>This study examined the prevalence, molecular characterization, and antimicrobial resistance of <i>Escherichia coli</i> associated with the bovine clinical mastitis in the northern part of Bangladesh. A total of 120 milk samples from infected cows in Dinajpur, Thakurgaon, and Nilphamari were collected. General bacteriological and biochemical tests were conducted to isolate and identify the bacteria. In addition, PCR assays were performed to molecularly detect and identify virulence and antibiotic resistance genes. Bacteriological, biochemical, and molecular methods confirmed the presence of <i>E. coli</i>, which constituted a total prevalence of 35.0%. Phylogenetic typing revealed two types, A1 (81.0%), and B2 (19.0%) (<i>p</i> < 0.001), suggesting that commensal strains predominating the total <i>E. coli</i> population. Virulence gene (<i>eae, stx1,</i> and <i>stx2</i>) screening was negative, indicating a lack of classical enteropathogenic/enterohemorrhagic characteristics. Antibiotic susceptibility testing showed that the level of antibiotic resistance was high, especially to ampicillin (100.0%), amoxicillin (92.86%), tetracycline (73.81%), erythromycin (61.91%), streptomycin (59.53%), and gentamicin (52.38%). The isolates were mostly sensitive to ceftriaxone (85.71%) and moderately sensitive to chloramphenicol and ciprofloxacin. The most abundant resistance gene was <i>tet</i>A (85.71%), followed by sul2 (59.52%), <i>blaTEM</i> (57.14%), <i>aadA1</i> (50.0%), <i>qnr</i> (23.80%), and <i>blaCTX-M</i> (19.04%) (<i>p</i> < 0.001), which are strongly connected to the phenotypic resistance pattern. In general, the results indicate that highly resistant <i>E. coli</i> are prevalent in the case of bovine mastitis, which highlights the necessity to use antibiotics reasonably, conduct more frequent monitoring, and use biosecurity to limit the spread of resistant pathogens in the dairy industry, as well as the environment.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"3499907"},"PeriodicalIF":2.3,"publicationDate":"2026-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13494350/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798269","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Heat Synchronization Protocols in Dairy Cattle (<i>Bos indicus</i>) and Success Factors in Niger.","authors":"Mahamadou Moussa Garba, Mani Mamman, Hamadou Issa","doi":"10.1155/vmi/4212869","DOIUrl":"10.1155/vmi/4212869","url":null,"abstract":"<p><p>Synchronization and induction of ovulation enable the control and harmonization of femaleestrus cycle facilitating artificial insemination (AI) by mitigating the challenges associated with heat detection. In Niger, progestogens alone or in combination with other hormones such asgonadotropin-releasing hormone (GnRH) and prostaglandin F2alpha (PGF_2α), are frequently employed to manage prolonged postpartum anestrus in cattle. This summary reviews their physiological mechanisms of action and practical application protocols. Additionally, complementary hormonal treatments notably PGF_2α and Equine chorionic gonadotropin (eCG) are commonly integrated intointravaginal device (IVD) based regimens. Understanding the hormonal dynamics of estrus is essential for maximizing the success rate of IVD-based synchronization program.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"4212869"},"PeriodicalIF":2.3,"publicationDate":"2026-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13479802/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148798450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dietary Inclusion of Casimiroa (<i>Casimiroa edulis</i> L) Seed Kernel Meal on Meat Qualities, Blood Profiles, and Cecal Microbiota of Broiler Chicken.","authors":"Mohammedjuhar Musa Adame, Mengistu Urge Letta, Biazen Abrar Bedru, Sileshi Gadissa Terfa, Meseret Girma Abebe, Misba Alewi Abdu","doi":"10.1155/vmi/2733198","DOIUrl":"10.1155/vmi/2733198","url":null,"abstract":"<p><p>The purpose of this study was to evaluate the effects of dietary inclusion of soaked Casimiroa seed kernel (CSK) meal on broiler meat quality, blood parameters, immunological response, and cecal microbiota. A total of 192 Cobb-500 broilers were assigned to four dietary treatments (0%, 1.5%, 3.0%, and 4.5% soaked CSK) in a completely randomized design with four replicates and 12 birds per replicate. The birds were fed isocaloric and isonitrogenous diets for 42 days. Majority of the optimal results were observed at the 1.5% and 3.0% soaked CSK inclusion levels, as evidenced by significant quadratic trends. Breast and thigh meat quality showed quadratic improvements in protein content, fat reduction, drip and cooking loss reduction, redness, and sensory scores (juiciness and tenderness). Key blood parameters (red blood cells, hemoglobin, total protein, immunoglobulins, and globulins) were improved quadratically, while serum triglycerides and cholesterol decreased linearly. Additionally, all soaked CSK inclusion levels quadratically reduced pathogenic E. coli and coliforms without impacting beneficial Lactobacilli and Enterococcus spp. Based on these results, dietary inclusion of soaked CSK meal up to 3.0% is a safe and effective strategy for simultaneously enhancing broiler meat physicochemical quality, improving metabolic and immune status, and promoting a beneficial gut microbiota balance.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"2733198"},"PeriodicalIF":2.3,"publicationDate":"2026-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13470872/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148766593","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of the Freezing Diluent Tonicity and Prefreezing Cooling Rates on Post-Thaw Sperm Quality Markers in the Pure-Bred Arabian Stallion.","authors":"Leili Mavalizadeh, Mohammad Reza Divar, Asghar Mogheiseh, Maryam Emami","doi":"10.1155/vmi/4438005","DOIUrl":"10.1155/vmi/4438005","url":null,"abstract":"<p><p>Stallion semen cryopreservation remains limited by inconsistent post-thaw sperm quality and fertility. This study evaluated the effects of freezing extender effective tonicity and prefreezing cooling rate on post-thaw sperm characteristics in pure-bred Arabian stallions. In a two-phase design, ejaculates (<i>n</i> = 40) from 10 stallions were processed using extenders with different osmotic conditions (330, 390, and 450 mOsm/kg) and subjected to cooling durations of 45 min, 1.5 h, or 3 h. Sperm quality was assessed using CASA, functional assays, flow cytometry, transcriptomic analysis, and a pilot fertility trial. Moderate hypertonicity (∼356 mOsm/kg effective tonicity) significantly improved sperm function, including higher proportions of live/nonapoptotic cells, enhanced mitochondrial activity, improved acrosome integrity, and reduced DNA fragmentation (<i>p</i> < 0.05). In contrast, excessive hypertonicity (∼383 mOsm/kg) increased necrosis and acrosome damage and tended to elevate the Bax/Bcl2 ratio although this effect was not statistically significant. Sperm kinematic parameters were not affected by osmolarity. Extending prefreezing cooling from 45 min to 1.5-3 h significantly improved motility, viability, mitochondrial function, acrosome stability, and DNA integrity, while reducing apoptosis and total cell death (<i>p</i> < 0.05), with no consistent additional benefit beyond 1.5 h. Oxidative stress (MDA) was not significantly affected. The optimized protocol resulted in pregnancy rates of 75.0% at Day 21 and 62.5% at Day 60. In conclusion, moderate hypertonicity combined with controlled slow cooling enhances stallion sperm cryosurvival and provides a practical strategy to improve fertility outcomes.</p>","PeriodicalId":23503,"journal":{"name":"Veterinary Medicine International","volume":"2026 ","pages":"4438005"},"PeriodicalIF":2.3,"publicationDate":"2026-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13470873/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148761491","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}