Vascular Wall Mesenchymal Stem Cells (VW-MSC) are vascular-resident stem cells with multipotent differentiation ability, contributing to vascular homeostasis and repair. Their secretome, including growth factors, cytokines, and extracellular vesicles, mediates paracrine effects on angiogenesis, immunity, and tissue regeneration. Given the pivotal role of VW-MSC in vascular repair and their sensitivity to environmental stressors, bioactive plant-derived compounds have emerged as promising modulators of their survival and regenerative potential. In the present study, we investigated the protective potential of microencapsulated extract of Vaccinium floribundum Kunth (VFM) berries on porcine VW-MSC exposed to serum deprivation induced cytotoxicity. Our results demonstrated that VFM extract did not alter pVW-MSC morphology but enhanced cell viability. Serum deprivation induced loss of cell adhesion, cell rounding, membrane blebbing, and reduced pVW-MSC viability by ~50%, whereas VFM treatment restored normal morphology and viability. Gene expression analysis revealed a significant decrease in caspase-3 expression, suggesting a possible attenuation of apoptosis. No significant changes in BAX, BCL2l1 or TP53 were detected. These effects are consistent with VFM bioactive compounds previously associated with cytoprotective and antioxidant activities in a model of porcine Aortic Endothelial Cells. Collectively, our results suggest that VFM supports pVW-MSC survival under stress conditions by modulating apoptosis and highlight its potential as a protective agent for stem cell-based regenerative and cardiovascular applications.

Rodent-associated viruses are important zoonotic agents of increasing relevance to public and veterinary health. Muroid rodents occupy diverse natural and human-modified environments, facilitating interactions with humans, livestock and wildlife. However, knowledge of rodent-associated viral diversity and the ecological processes influencing viral maintenance and transmission remains fragmented. This review synthesizes published literature on viral diversity, host-virus associations, research trends, and ecological drivers of viral occurrence in Asian muroid rodents. PubMed and Scopus were systematically searched using terms related to rodents, viruses, and Asian regions. Extracted data included publication characteristics, diagnostic methods, host and virus taxonomy, geography, and ecological metadata. Descriptive analyses, geospatial mapping, and network approaches were applied to identify patterns and knowledge gaps. Of 1,049 records retrieved, 165 studies published between 1968-2025 from 30 Asian countries met the inclusion criteria. These studies documented 106 viral taxa representing 74 genera and 35 families. Research activity increased markedly after 2000 alongside advances in molecular diagnostics and metagenomic sequencing. Studies were concentrated in East and mainland Southeast Asia. Orthohantavirus spp. were the most studied viral group, whereas many other viruses remained poorly characterized. Network analyses identified core murine genera, particularly Rattus, Apodemus, Niviventer, Bandicota and Mus, sharing extensive viral associations. Host age, sex, population density, habitat disturbance, climate, and biodiversity were commonly linked with viral prevalence and dynamics. Despite substantial progress, gaps remain in ecological integration, longitudinal surveillance, and comparative study design. Future research should prioritize standardized, ecologically informed surveillance within an integrated One Health framework to improve zoonotic risk assessment and prevention.

Toxoplasma gondii is an important zoonotic parasite affecting humans and livestock, and reliable serological tools are needed for disease surveillance in small ruminants. This study developed and evaluated a recombinant T. gondii matrix antigen 1 (TgMAG1) based indirect ELISA (iELISA) for detecting anti T. gondii antibodies in goat sera from northeastern Thailand. A truncated TgMAG1 fragment (amino acids 31-93) was selected to minimize cross-reactivity with Neospora caninum, expressed in Escherichia coli, and confirmed by SDS-PAGE, western blotting, and mass spectrometry. Overall, 213 goat sera were tested using latex agglutination test (LAT), indirect fluorescent antibody test (IFAT), and the TgMAG1-based iELISA. Seroprevalence was 34.7% by LAT and 38.5% by IFAT. Using LAT as the comparator, the iELISA showed 90.5% relative sensitivity (rSe; 95% CI: 81.7-95.3%), 85.6% relative specificity (rSp; 95% CI: 78.8-90.5%), and an area under the curve (AUC) of 0.91 (Cohen's kappa [κ] = 0.73); using IFAT, rSe was 92.7% (95% CI: 84.9-96.6%), rSp was 66.4% (95% CI: 58.0-73.9%), and the AUC was 0.85 (κ = 0.54). These findings suggest the TgMAG1-based iELISA offers high rSe for detecting T. gondii exposure in goats, though further validation with broader serum panels is warranted to confirm its rSp.

Chlamydia psittaci (C. psittaci) is a major intracellular pathogen of pigeons that often requires prolonged antimicrobial therapy; however, high-dose oxytetracycline (OTC) may increase toxicity. Glycyrrhizic acid (GA), a bioactive licorice constituent, has anti-inflammatory, antioxidant, and antimicrobial-enhancing properties. This study evaluated whether GA improves the pharmacokinetics, therapeutic efficacy, and safety of OTC against experimental C. psittaci infection in pigeons. C. psittaci was isolated and confirmed via embryonated egg inoculation and real-time PCR. Eighty-four pigeons were allocated to seven groups: negative control (G1), infected control (G2), oral OTC (G3), intravenous OTC (G4), oral GA (G5), oral OTC plus GA (G6), and intravenous OTC plus GA (G7). Pharmacokinetics were assessed using a validated green HPLC-UV method, while antimicrobial efficacy, inflammatory and oxidative stress markers, and hepatic and renal biomarkers were evaluated. GA showed borderline synergism with OTC (fractional inhibitory concentration index, 0.499) against the tested isolate. The HPLC method showed excellent analytical performance, with an AGREE greenness score of 0.82. Compared with OTC alone, GA significantly (P < 0.05) increased Cmax and AUC0-24, improved absolute oral bioavailability from 29% to 38%, increased relative bioavailability 1.5-fold, and reduced apparent oral and systemic clearance. Combination therapy markedly reduced chlamydial burden and inclusion body formation, attenuated IL-6, TNF-α, and malondialdehyde levels, and restored glutathione and hepatic and renal biomarkers compared with monotherapies. These findings support GA as a promising adjunct to OTC therapy for avian chlamydiosis, enhancing OTC exposure and therapeutic efficacy while reducing infection-associated tissue injury, potentially mitigating adverse effects associated with prolonged OTC therapy.

Bovine leukemia virus (BLV) infection in calves may contribute to early dissemination within dairy herds. This study aimed to determine the BLV infection prevalence in pre-colostral and neonatal calves and its association with management-related factors in dairy farms in Uruguay. Two studies were conducted. Study 1 was a retrospective analysis of archived pre-colostral blood samples from 75 calves from one farm. Samples were analyzed by quantitative real-time PCR (qPCR) and ELISA, and previous ELISA and qPCR results from dams of calves positive by qPCR and/or ELISA were reviewed. Study 2 was a cross-sectional study of blood samples from 542 neonatal calves aged 1-30 days from 30 dairy farms, analyzed by qPCR, with management data collected by questionnaire. In Study 1, 6.7% (5/75) of calves were qPCR-positive, and 4.0% (3/75) were ELISA-positive; one calf was positive by both assays. All seven dams of calves positive by qPCR and/or ELISA were ELISA-positive, while five were also qPCR-positive. In Study 2, 8.9% (48/542; farm-level bootstrap 95% CI 3.7-15.1%) of calves were qPCR-positive, within-herd apparent prevalence ranged from 0% to 54.5%, and 43.3% (13/30) of farms had ≥1 qPCR-positive calf. In Study 1, qPCR positivity before colostrum ingestion was consistent with prenatal transmission. Exploratory multivariable analysis suggested possible farm-level associations between herd size, maternal vaccination against neonatal calf diarrhea, rotation of calf-rearing areas, and type of liquid feed and qPCR-positive calves. These findings suggest that BLV infection can be detected during the neonatal period and support investigation of management and biosecurity practices.

Mammary gland tumors (MGTs) are common in female dogs, and are usually not treated with chemotherapy due to the lack of evidence of benefit for the patients. In clinical practice, when a female dog receives neoadjuvant or adjuvant chemotherapy, minimal improvement is achieved. Therefore, chemotherapy is not usually applied for these patients. We hypothesized that some canine MGTs do not respond to chemotherapy due to intrinsic chemoresistance. Chemoresistance can be a challenge and is often associated with overexpression of ABC transporters, such as MDR1, MRP1, MRP2, MRP3 and BCRP, which reduce the intracellular accumulation of chemotherapeutic agents into the cells. This study aimed to induce chemoresistance in canine MGT cells in vitro, identify potential genetic changes linked with resistance and to evaluate the expression of ABC transporter genes (MDR1, MRP1, MRP2, MRP3 and BCRP) using RT-qPCR. We also aimed to verify the relation between chemoresistance and single nucleotide polymorphisms (SNPs). Five cell cultures were used (UNESP-CM1, UNESP-CM4, UNESP-CM5, UNESP-CM70 and UNESP-CMR1). In four cell lines, chemoresistance was induced with doxorubicin (DOX) and one cell line was naturally resistant. Single Nucleotide Polymorphism (SNP) analysis via genotyping and RT-qPCR were performed in cell line cultures, before and after chemoresistance induction. Evaluation by RT-qPCR showed that the UNESP-CMR1 cell presented higher expression of MDR1, BCRP, MRP1 and MRP3 compared to the other cells. Three cells lines treated with DOX had increased expression of MDR1, MRP1 and MRP2 after treatment. SNP analysis identified 35 potentially altered SNPs, with emphasis on SNPs on chromosomes 17 and 24, associated with the ALK and RALGAPA2 genes. Interestingly, ALK gene presents a tyrosine kinase domain and it is considered a therapeutic target. In conclusion, these preliminary data suggest that the induction of chemoresistance in canine mammary carcinoma cells may be associated with variations in the expression of ABC transporter genes.

Concentrates are typically fed twice daily to supplement forages in horses to facilitate management, especially those with higher energy needs, but high-grain meals can contribute to digestive and metabolic challenges, including insulin dysregulation. This study investigated the effects of feeding frequency on glycemia, insulinemia, and insulin sensitivity measured by an oral sugar test in moderately exercised young horses. Thirty-two healthy Quarter Horses (2-3 years old) were assigned to either traditional feeding (TF; two grain meals/day) or fractioned feeding (FF; automated grain delivery every hour for 10 h/day), with grain comprising 40% of daily dry matter intake and forage 60%. Oral sugar tests (OST) were conducted at baseline and after 30 and 60 days of treatment. On day 45, a subset of horses (n = 10) completed an 8-h postprandial glucose and insulin assessment; plasma basal cortisol was also measured at 0, 30 and 60 days. OST results showed elevated 75-min insulin on day 30 for both treatments (P < 0.001). During the 8-h postprandial test, TF horses exhibited higher glucose and insulin peaks than FF horses(P < 0.030). Area under the curve for insulin (AUC) was higher (P = 0.032) in TF than FF at 60 and 180 mins, indicating greater postprandial insulinemia with larger, less frequent meals. Total AUC for insulin was lower (P = 0.006) in FF horses (1119.04 ± 102.30 μIU·min/mL) compared to TF horses (1406.00 ± 160.8 μIU·min/mL). These findings suggest that fractioned feeding attenuates postprandial insulin responses due to a lower non-structural carbohydrate (NSC) load per meal and therefore warrants direct testing when managing horses at risk for insulin dysregulation or hyperinsulinemia-associated laminitis. This finding could be another tool to manage insulin dysregulated horses.

Fasciolosis is a food-borne disease affecting livestock worldwide. Improved diagnostic methods for detecting fasciolosis have been introduced; however, limited research has compared these methods. The current study aimed to compare the detection of Fasciola in naturally infected slaughtered dairy cattle using four methods. Blood and faecal samples were collected from slaughtered cattle in three high-throughput abattoirs in the Eastern Cape Province, South Africa, from 2020 through 2021. Post-mortem liver examination was conducted to detect and identify Fasciola spp. Blood from slaughtered carcasses was analyzed using an antibody ELISA to detect Fasciola antibodies, while faecal sedimentation and quantitative real-time PCR (qPCR) were used to detect Fasciola eggs and Fasciola DNA, respectively, in faeces. A binomial Generalized Linear Model (GLM) and Cohen's Kappa test were used to compare the detection rate and determine the level of agreement across the four diagnostic methods. Significant differences (P = 0.0001) across the four diagnostic methods were observed. The highest detection rate was observed using the post-mortem liver examination (94.5%), followed by qPCR (90.4%), ELISA (22.6%), and faecal sedimentation (3.53%) in that order. There was a poor agreement between sedimentation and Ab-ELISA (kappa = -0.105, p = 0.128), sedimentation and post-mortem liver examination (kappa = -0.054, p = 0.163), and qPCR and liver examination (kappa = -0.076, p = 0.341). In conclusion, results from the current study revealed significant differences among the four methods in the detection of Fasciola spp. post-mortem liver examination and qPCR were more reliable, sensitive, and accurate than ELISA and sedimentation methods.

Dogs with oliguric acute kidney injury (AKI) of infectious etiology are frequently managed with renal replacement therapy (RRT) and β-lactam antibiotics. Ampicillin and sulbactam are small, hydrophilic, renally eliminated molecules susceptible to extracorporeal clearance during RRT, yet the magnitude of their extracorporeal removal during RRT has not been quantified in dogs. Nine client-owned dogs with oliguric AKI of IRIS grade IV or greater that received ampicillin/sulbactam (22 mg/kg IV q8h) prior to undergoing low-efficiency hemodiafiltration (LE-HDF). Prospective, single-center observational study. Blood samples were collected immediately before and after the first LE-HDF session. Plasma ampicillin and sulbactam concentrations were quantified by validated LC-MS/MS and compared using a Wilcoxon signed-rank test; the relationship between pre- and post-dialysis concentrations was also characterized. Concentrations were interpreted against published minimum inhibitory concentration (MIC) thresholds for E. coli and Leptospira spp. LE-HDF reduced median ampicillin concentrations from 81.2 μg/mL (Interquartile range (IQR) 68.2-103.9) to 19.5 μg/mL (IQR 15.2-41.5), a median reduction of 75.6% (geometric mean ratio 0.23, 95% confidence interval (CI) 0.19-0.29), and sulbactam from 64.6 μg/mL (50.6-108.5) to 18.7 μg/mL (15.4-42.4; reduction 73.2%, geometric mean ratio 0.29, 95% CI 0.22-0.38). Post-dialysis ampicillin and sulbactam concentrations were strongly correlated with pre-dialysis concentrations (Spearman's ρ = 0.93 and 0.75, respectively). Despite this marked reduction, post-dialysis ampicillin concentrations exceeded all relevant MIC thresholds in all 9 dogs. A single LE-HDF session markedly reduces ampicillin and sulbactam concentrations in oliguric dogs, yet supratherapeutic baseline exposure maintained pharmacodynamic target attainment post-dialysis.

The routine removal of seminal plasma during processing of canine semen remains controversial. This study compared standard centrifugation (750 ×g, 5 min, 22 °C), cushion centrifugation (1000 ×g, 20 min, 22 °C), and syringe-filter filtration (Millex-GP 0.22 μm, retrograde flushing with 1 ml TRIS-citric acid-fructose buffer) for their effects on canine semen quality. Sperm concentration, sperm recovery rate, total motility, progressive motility, membrane integrity, morphology and kinematics were evaluated. All semen processing methods resulted in a considerable sperm loss with sperm recovery rate being less reduced after standard centrifugation relative to cushion centrifugation and filtration (Standard 80.0 ± 3.1[a]; Cushion 55.1 ± 3.1[b]; Filter 59.8 ± 4.1[b] %; ab: p < 0.001). Sperm concentration was considerably reduced after cushion centrifugation and filtration (Control 409.4 ± 27.9[a]; Standard 738.6 ± 69.1[ac]; Cushion 275.4 ± 29.8[ad]; Filter 229.0 ± 34.8[b] x 10[6]/ml; ab, cd: p < 0.01). Total and progressive motility were significantly decreased after filtration vs. both centrifugation methods. Membrane integrity was most reduced in filtered semen, standard centrifugation caused a numerically minor but significant decrease in membrane integrity vs. control and cushion centrifugation. Filtration decreased kinematic parameters in comparison to standard and cushion centrifugation. Filtration significantly increased the percentage of morphologically abnormal sperm that did not differ among control, standard centrifugation and cushion centrifugation. In conclusion, processing methods investigated in this study decreased semen characteristics compared to raw semen of healthy dogs. Effects of filtration clearly exclude the here tested method from further use and investigations in dogs. Cushion centrifugation offered limited benefits, but without improvement of sperm recovery by modifications of the method its use is not of interest in commercial dog semen freezing.

This preliminary, exploratory study evaluated animal welfare in four structurally different sheep housing systems in Eastern Anatolia, Türkiye, under cold winter conditions, using the AWIN welfare protocol. A total of 1920 observations were collected across the four systems during six weekly visits, covering body condition score (BCS), fleece contamination and loss, hoof length, lesions, lameness, mucous membrane colour, eye-nasal discharge, faecal contamination, drinking-water cleanliness, behavioural tests, stocking density, and lamb mortality. Comparisons used Firth bias-reduced logistic regression with cluster-bootstrap P-values to account for the repeated-visit sampling structure. Enclosed concrete-wall housing with a wooden slatted floor (CC-WB) showed the most favourable overall welfare profile, with the highest BCS, lowest lamb mortality, and significantly better lesion scores and mucous membrane colour than the traditional stone-walled system (CS-SB); however, its BCS remained below the adequacy threshold and its water hygiene was inferior to CS-SB, so it is best regarded as the least inadequate rather than an optimal system. Enclosed concrete-wall, concrete-based housing (CC-CB) showed the most negative, statistically robust profile, with significantly higher fleece loss, lameness, and body lesions; it also had the lowest stocking density, so flooring type and space allowance could not be disentangled. Because each housing system was represented by a single commercial farm and the design was exploratory, housing type was associated with, but cannot be shown to cause, the differences observed; these findings are preliminary and hypothesis-generating, and larger, replicated, multi-farm studies are needed before they inform practical adoption of any housing design.

Antimicrobial resistance (AMR) in Staphylococcus spp. associated with canine otitis externa (COE) represents an increasing challenge for veterinary practice and public health. We conducted a systematic review and meta-analysis following PRISMA guidelines to estimate global AMR prevalence in Staphylococcus spp. from COE and to examine variation by antimicrobial class, agent, species, and continent. A literature search was conducted in four electronic databases to identify observational studies reporting antimicrobial susceptibility of Staphylococcus isolates from COE. Twenty-eight studies published between 1995 and 2024, comprising 22,864 isolates, met the inclusion criteria. Random-effects meta-analyses were performed. The overall pooled AMR prevalence among Staphylococcus spp. was 33.83% with high heterogeneity. Beta-lactams showed the highest pooled resistance (60.32%; 95% CI: 47.85,71.58), with a significant difference between methicillin resistance and third-generation cephalosporin resistance. Species-level analyses revealed significant interspecies differences for aminoglycosides and fluoroquinolones. Continent-level analysis revealed variation in specific antibiotic classes (p < 0.01), with resistance consistently peaking in South America and remaining lowest in Africa, whereas beta-lactam resistance showed striking global uniformity (p = 0.70). Meta-regression showed a significant positive association between publication year and AMR (QM = 18.95, p < 0.0001; coefficient = 0.0561 per year). In conclusion, our findings indicate that AMR among Staphylococcus spp. from COE is substantial, varies by class and species, and shows an increasing temporal trend alongside marked regional heterogeneity. These findings underscore the need for local surveillance networks, strengthened antimicrobial stewardship, standardized diagnostic protocols, and a One Health approach to mitigate AMR risks to animal and human health.

Enzootic bovine leukosis, paratuberculosis, and bovine tuberculosis are chronic infectious diseases with important implications for dairy productivity and animal health. Evidence integrating the three agents into the same animals is scarce in tropical highland dairy systems and almost absent for herds enrolled in official bovine tuberculosis certification programs. An analytical observational study evaluated 733 cattle from 37 specialized dairy herds in a highland dairy region of Colombia using serological (ELISA) and molecular methods (qPCR, ddPCR) to detect bovine leukemia virus (BLV), Mycobacterium avium subsp. paratuberculosis (MAP), and Mycobacterium bovis. Mixed-effects logistic regression with a random intercept for herds accounted for the clustering of animals within herds, and agreement between diagnostic tests was quantified using Cohen's kappa. At the herd level, BLV was detected in 70.3% of the herds by ddPCR and 67.6% by ELISA, MAP was detected in 27.0% by ELISA and 18.9% by fecal qPCR, and M. bovis was not detected in any herd. At the animal level, BLV proviral DNA was detected in 34.8% of the cattle, and BLV antibodies were detected in 31.9%; MAP fecal DNA was detected in 1.2%, and MAP antibodies were detected in 1.9%, and no animal tested positive for M. bovis by any method. Agreement between the two BLV definitions was substantial (kappa = 0.79), whereas the two MAP definitions identified essentially nonoverlapping animals (kappa = 0.07). The odds of MAP fecal shedding did not differ between BLV-positive and BLV-negative animals, but the estimate was imprecise and compatible with a wide range of effects (OR = 0.26; 95% CI: 0.03-2.43; p = 0.24, from the mixed-effects model), reflecting only nine MAP-positive animals and a single coinfected animal. Age was associated with BLV proviral detection in the mixed-effects model, although the association was less precise and not statistically significant when cluster-robust standard errors were used. BLV was detected in a high proportion of animals and herds, whereas MAP detection was limited, and no evidence of current M. bovis infection was detected using the diagnostic approaches applied in this selected population. The absence of a detectable BLV-MAP association does not imply biological independence, given the small number of MAP-positive animals. The marked clustering of BLV detection within herds (intraclass correlation coefficient of 0.45) indicates substantial unmeasured herd-level heterogeneity and has direct implications for the design of surveillance in these systems. These findings support pathogen-specific surveillance and control strategies.