Astragalus species are major causes of livestock poisoning, traditionally attributed to swainsonine (SW). This study compared Astragalus pehuenches and Astragalus illini to determine whether cytotoxicity differences could be explained solely by SW or might involve other constituents. Alkaloid-enriched extracts (AEEs) were evaluated using phytochemical analyses and SW-normalised cytotoxicity assays, followed by A. illini extract fractionation. SW was quantified by HPLC-MS/MS, selenium and nitro compounds were assessed, and saponin-equivalent content was estimated. Although A. pehuenches contained more SW than A. illini (4.70 vs. 1.49 μg SW/mg AEE), the A. illini extract caused a greater reduction in C6 glioma viability under SW-normalised exposure. CC50 values were 802.79 and 447.74 μM SW equivalents for A. pehuenches and A. illini AEEs, respectively. Neither extract induced detectable LDH release. Qualitative AO/EB and lectin staining showed changes in acidic vesicular staining patterns and intracellular glycoconjugate staining patterns consistent with SW-associated cellular alterations. However, because equivalent SW exposure required different total extract concentrations, their intrinsic cytotoxic potencies could not be directly compared. Fractionation of the A. illini extract showed that cytotoxicity did not parallel SW concentration, while some of the most cytotoxic fractions contained abundant saponin-related compounds. Non-SW metabolites, potentially including saponins, may be associated with the observed cytotoxicity; however, their specific contributions remain unconfirmed. These findings support integrated phytochemical and biological analyses for investigating cytotoxicity associated with constituents other than SW in Astragalus extracts. Future studies should structurally characterise candidate compounds and test their individual effects to establish their causal roles in the observed cytotoxicity.

Conspecific social competence - the capacity of domestic dogs to engage in contextually appropriate interactions with other dogs - is a clinically significant yet systematically underaddressed construct in canine behavioural science. Existing approaches to reactivity and fear-based behaviour reduce problematic responses effectively, but are oriented toward managing conspecific exposure rather than developing genuine communicative and regulatory competence, and no framework currently exists for developing these capacities in adult dogs who have passed the early socialisation window. This paper proposes a definition of conspecific social competence, distinguishing it from human-directed social competence and from the absence of reactive behaviour, and presents the Canine Social Competence Development Framework. Each component was retained only where it could be grounded in published findings on canine social learning, communication, or welfare (Pongrácz et al., 2001; Bradshaw and Rooney, 2016; Fugazza et al., 2018), while its formulation as a distinct and applicable component of practice draws on clinical observation. The framework identifies one foundational prerequisite - allowing direct conspecific contact - and three interdependent pillars: experiencing social consequences, interacting with conspecifics of varying behavioural profiles, and minimising human interference in interaction dynamics. Clinical implications are discussed, including partner selection, session structure, indicators of progress, and the management of safety risks inherent in facilitating direct conspecific contact. The limitations of the framework are acknowledged, including its current reliance on clinical observation rather than empirical testing, and directions are outlined for the empirical validation it requires.

The trail plate is a modular system consisting of a trail bar and synthesis modules made of stainless steel, originally developed for humans. This study aimed to evaluate the applicability and effectiveness of a modified trail plate for the treatment of long bone fractures in dogs. Initially, the trail plate was designed by using FEA to set the final model. Three trail plate sizes were designed according to corresponding locking screw diameters. The designs were created using computer-aided design software, and 3D models were produced from the commercial Ti-6Al-4 V powder via direct metal laser sintering (DMLS). The trail plate application included radius/ulna (n = 5), femur (n = 7), and tibia/fibula (n = 7). Radiographic bone healing occurred within 60 days in 100% of radius/ulna fractures, within 90 days in 70% of tibia/fibula fractures, and between 30 and 120 days postoperatively in femoral fractures. Radius/ulna fractures exhibited minimal callus formation, and all animals were free of lameness by 60 days. Femoral fractures showed mostly moderate callus formation, with lameness resolved in 57% of dogs by 60 days. In tibia/fibula fractures, callus ranged from minimal to abundant, with lameness resolved in six dogs by 60 days. Complications occurred in 36.8% of cases, primarily due to trail bar displacement of the module; only one case involved implant failure. In conclusion, this preliminary case series suggests that modified DMLS-manufactured titanium trail plate was clinically feasible for selected canine diaphyseal fractures. However, fixation-related complications indicate that further design optimization and controlled biomechanical studies are necessary before broader recommendation.

Acute Hepatopancreatic Necrosis Disease (AHPND) is one of the most destructive bacterial diseases in shrimp aquaculture and is caused by virulent Vibrio parahaemolyticus strains carrying the pVA1 plasmid encoding the binary toxins PirA and PirB, resulting in severe economic losses and threatening sustainable production worldwide. Although lytic bacteriophages have emerged as promising alternatives to antibiotics because of their specificity and self-amplifying properties, their long-term effectiveness is constrained by the rapid evolution of bacterial resistance through multiple mechanisms, including surface receptor modification. Recent evidence indicates that membrane vesicles (MVs) can modulate phage-bacterium interactions through mechanisms that vary among bacterial systems. In Bacillus subtilis, MVs can transfer phage attachment molecules and transiently restore phage sensitivity, whereas in Vibrio parahaemolyticus, MVs have been reported to enhance phage efficacy, although the underlying mechanism remains unresolved. This review synthesizes current knowledge on AHPND epidemiology, phage infection and resistance, MV biology, phage cocktail design, formulation, and delivery strategies. It further evaluates the potential of MV-assisted phage approaches as a complementary strategy for sustainable AHPND management, while highlighting the need to validate functional receptor restoration in resistant Vibrio and address challenges in MV standardization, biosafety, and field application.

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.