PAMK are heterogeneous glycans with immunomodulatory potential, but their structural basis and mechanisms in regulating thymic injury remain unclear. Structural characterization revealed that PAMK is mainly composed of arabinose, galactose, rhamnose, galacturonic acid, and glucose, featuring a dominant backbone consisting of →4)-α-D-GalpA-(1→, →4)-β-D-Galp-(1→, →2)-α-L-Rhap-(1→, and →3,4)-α-D-GalpA-(1→ with highly branched side chains. This complex structure provides the biochemical foundation for its bioactivity. To explore its immunoprotective effects, we established a CTX-induced thymic necroptosis model in goslings. CTX markedly disrupted cytokine profiles and activated necroptosis signaling, accompanied by downregulation of novel-miR-2 and upregulation of TRADD and MLKL. Dietary PAMK supplementation significantly restored cytokine homeostasis and inhibited necroptosis pathway activation. Mechanistically, PAMK upregulated novel-miR-2, which negatively regulated TRADD and MLKL expression, thereby attenuating necroptosis signaling. Overexpression of novel-miR-2 suppressed TRADD/MLKL-mediated necroptosis, whereas its inhibition produced the opposite effect. Furthermore, PAMK mitigated oxidative stress and necroptotic cell death induced by TRADD and MLKL overexpression. Collectively, our findings demonstrate that the defined structural features of PAMK underpin its ability to protect against CTX-induced thymic necroptosis by upregulating novel-miR-2 and suppressing the TRADD/MLKL axis. This work links PAMK's composition to its immunoprotective function and highlights its potential as a safe immunoregulatory agent in poultry.
While dogs' behavioural indicators of psychological stress are widely used, their function as reliable stress markers remains debated. Using the Strange Situation Procedure (SSP) as a social-stress paradigm, we evaluated contextual correlates of dog behaviours to explore their validity as stress indicators. In 64 dogs, behaviours commonly used as stress markers were video-coded and analysed via GLMMs to investigate differences across test episodes and attachment styles (secure, anxious, avoidant), while salivary cortisol, oxytocin, and rectal temperature provided a physiological framework. Episode had significant effects on shaking (p < 0.05), lip-licking (p < 0.001), paw-lifting (p < 0.001), whining (p < 0.001), head-turning (p < 0.001), sniffing the door (p < 0.001), and locomotion (p < 0.001), while attachment style significantly affected head-turning only (p = 0.004). Our findings suggest that the observed behaviours were not expressed linearly in relation to expected stress levels across the SSP episodes. This lack of linearity, combined with the divergent physiological correlates found across attachment groups-for instance, oxytocin correlated positively with lip-licking in securely attached dogs (p < 0.05), but negatively in anxious ones (p < 0.05) -, suggests that these behaviours do not function as simple stress indicators. Rather, their meaning appears to be contingent upon the individual's internal state and the socio-environmental context. We discuss our results in light of an alternative semiotic interpretative framework.
Significant causes of morbidity in veterinary medicine are tumourigenic diseases and pathogenic infections, which pose significant threats to animal health. Traditional methods for treating tumours and bacterial infections primarily involve chemotherapy and antibiotic therapy, both of which are associated with drug resistance and adverse effects. Photodynamic therapy (PDT) is gaining increasing interest in veterinary clinical practice because of its distinctive benefits such as spatiotemporal selectivity, low toxicity, fewer negative effects, and reduced risk of drug resistance. This review summarises the fundamental principles of PDT and its applications in combating tumours and microbial infections in veterinary practice involving companion animals, livestock, wildlife, and reptiles. It is worth noting that the existing clinical evidence is limited by very small sample sizes and the absence of comparator groups; however, these studies have provided a preliminary theoretical foundation and technical reference for further clinical development of PDT. Moreover, the opportunities and challenges associated with PDT in veterinary clinical practice are discussed herein.
AIM (apoptosis inhibitor of macrophage), also known as CD5L, is a macrophage-derived scavenger protein with broad immunomodulatory roles in mammals. Secreted by tissue macrophages, AIM circulates bound to IgM pentamers, which stabilize the protein in blood; free AIM is released during inflammation to influence immune signaling and cellular homeostasis. (Yang et al. 2023). In cats, AIM exhibits distinctive structural and biochemical properties, existing as both a three-domain (37 kDa) and a four-domain (45 kDa) variant generated through exon 3 duplication (Evangelista et al. 2025), and binding IgM approximately 1000-fold more tightly than murine AIM due to a species-specific positively charged cluster in its third SRCR domain (Miyazaki et al., 2018). These differences suggest feline AIM may function distinctly from human/mouse AIM. Feline injection-site sarcomas (FISS) arise in the context of chronic inflammation and frequently contain abundant tumor-associated macrophages (TAMs)associated with tumor aggressiveness and progression (Gomes et al., 2025). Given AIM’s macrophage origin and its established roles in inflammatory regulation, autophagy signaling, and complement-mediated tumor targeting, we propose a directional hypothesis: that the high-affinity IgM-binding of feline AIM constrains free AIM availability in vivo, shifting the functional balance away from complement-mediated tumor cytotoxicity and toward macrophage-intrinsic M2-polarizing pathways, thereby promoting an immunosuppressive tumor microenvironment that contributes to FISS progression. Exploring AIM in this context may provide new perspectives for investigation in feline tumor immunology.
Veterinary forensic science is a specialized discipline that investigates non-natural animal deaths and injuries. Determining the cause of death in free-roaming cats often presents challenges due to underdeveloped pattern recognition methods and the intricate interplay of events and contributing factors. The integration of natural language processing into forensic investigations may offer potential for the computerized automation of document analysis, with the goal of supporting investigational decision-making. The objective of this study was to develop an AI framework leveraging a transformer-based large language model (LLM) to classify veterinary forensic autopsy reports derived from 271 feline autopsies and to identify the causes of death. We first identified discriminative lexical patterns between vehicular trauma (VT; n = 111) and non-VT cases (n = 160) using Term Frequency-Inverse Document Frequency analysis. The best-performing model, based on Clinical-Longformer, achieved an accuracy of 0.945, precision of 0.885, recall of 1.000, F1 score of 0.939, and AUC-ROC of 0.950. Model interpretability using Integrated Gradients revealed that VT cases were characterized by terms associated with traumatic physical injuries, whereas non-VT cases exhibited a broader range of diagnostic descriptions. These findings demonstrate the feasibility of applying LLMs to veterinary forensic pathological texts and underscore the importance of domain-specific token interpretation for model explainability.
This study compared liquid chromatography-mass spectrometry (LC-MS), micro-bioassay (MBa), and enzyme-linked immunosorbent assay (ELISA) for quantification of lincomycin (10 mg/kg BW) in porcine serum and evaluated lincomycin pharmacokinetic (PK) data in healthy piglets also co-infected with Actinobacillus pleuropneumoniae and Pasteurella multocida. All methods satisfied standard bioanalytical validation criteria, including low limits of detection and quantification, acceptable intra-assay imprecision and error, and strong linearity across clinically relevant concentration ranges. Serum concentrations measured by MBa and ELISA showed high correlation with LC-MS in both healthy and infected models (Pearson r ≥ 0.97, low bias on Bland-Altman analysis); they produced comparable PK parameters (Cmax, Tmax, area under the curve [AUC], clearance, and mean residence time), indicating suitability for PK/pharmacodynamic (PD)-based dose optimization. PK analysis demonstrated rapid absorption after intramuscular administration and disease-associated alterations in selected distribution and disposition parameters. AUC and clearance were nearly similar across analytical methods and largely consistent between software platforms (WinNonlin and PKSolver). Thus, lower-cost MBa and ELISA-combined with freely available PKSolver-can generate robust, decision-relevant PK/PD data for lincomycin in swine, providing a practical framework for antimicrobial dose refinement and stewardship in resource-limited veterinary and farm settings.
Current vaccines against paratuberculosis offer incomplete and variable protection that raises questions on the ruminant immune responses to Mycobacterium avium subspecies paratuberculosis (Map) infection, particularly at the mucosal level. Thus, this study examined the immune responses established in the jejunum of goats vaccinated with oral, intradermal, or subcutaneous inactivated vaccines 11 months post-vaccination and 10 months after experimental Map challenge. In general terms, the mucosal response of non-challenged animals was not affected by vaccination. However, the quantification of immune cell populations, cytokine expression, recall responses of tissue-isolated mononuclear leukocytes, and secretory IgA levels in vaccinated and challenged animals with focal lesions suggested a local Th1 polarization. In contrast, those with diffuse lesions exhibited features compatible with the early development of a more Th2-biased response. The vaccination route played a lesser role in the immune response than lesion severity. Orally vaccinated and challenged goats, showed reduced local IL-1β and IL-17A expression, which may explain the milder lesions observed in these animals, despite their high bacterial burden, as these cytokines participate in macrophage recruitment, granuloma formation, and bacterial clearance. Nevertheless, owing to the high variability among challenged animals, larger sample sizes would allow robust conclusions to be drawn in further studies.
In this study, Salmonella Typhimurium (ST) vaccine diluent effects on caecal tonsils gene regulation in layer chicks were investigated. A live attenuated ST vaccine was reconstituted in Marek's and/or water diluents, chicks vaccinated and caecal tonsils were assessed for global gene expression at days 7 and 14 post-vaccination. Differentially expressed genes (DEGs) involved in the activation of the intestinal immune network for IgA production were upregulated earlier (e.g. day 7 post-vaccination) in the water diluent group, while in the Marek's diluent group, this pathway was enriched by the upregulated DEGs at day 14 post-vaccination. The commonly upregulated biological pathways between the two diluents were cytokine-cytokine receptor interaction, toll-like receptor signaling pathway and cytosolic DNA-sensing pathway. At day 7 post-vaccination, 28.5% and 40.2% of the significantly upregulated genes were unique to Marek's and water diluents, respectively, whereas 31.3% were common between the Marek's and water diluent groups, showing the role of the diluent in priming the vaccine for host immune system modulation. At day 14 post-vaccination, 52.7% and 15.4% of the upregulated DEGs were unique to Marek's and water diluents, respectively. Overall, the data showed that the live attenuated ST vaccine reconstituted in Marek's diluent affected the regulation of immune system-related genes in the caecal tonsils of chicks, with a difference in pattern when water was used as a diluent.
Prohibitive costs associated with high-field MRI systems have resulted in reduced access in resource-limited areas with consequential healthcare inequities. This has renewed interest in purposefully designed low-field (LF) systems that, despite inherently lower signal-to-noise ratios, have advantages in terms of cost, portability, and accessibility. This pilot study successfully translated a previously developed LF (0.05 T) MRI canine cadaver brain protocol for in vivo application and compared the acquired images with paired 1.5 T images from 21 different canine patients. The visibility of 15 anatomic features was ordinarily scored (scale 1-3) on transverse T1-weighted post-contrast (21/21 patients) and T2-weighted (17/21 patients) sequences. Lateral ventricles were easily visualized (89.3%-100% scored 3/3) across all sequence‒system combinations, with T2-weighted sequences also providing good identification of the mesencephalic aqueduct (56.3%-100%), fourth ventricle (77.1%-100%), and thalamus (68.8%-100%). Absolute visual grading characteristics (VGC) analysis confirmed comparable performance of the LF system to the 1.5 T MRI for these features, a particularly relevant finding given the system's originally intended application for pediatric hydrocephalus neuroimaging in resource-limited areas. This study represents the first in vivo usage and evaluation of a 0.05 T MRI in a clinical veterinary context.
The objective was to evaluate the potential of machine learning algorithms utilizing thermal imaging and/or milk-based parameters for the identification of subclinical mastitis(SCM) in dairy cows. Holstein dairy cows (n = 194) were enrolled, and milk samples were collected from each quarter separately (n = 776 quarter-level samples) and analyzed for milk-based parameters. Four images per cow, representing all four quarters, were captured using a handheld infrared camera. Skin temperatures were extracted using Fluke Connect software, yielding 776 images for analysis. SCM was defined at the quarter level as SCC > 200,000 cells/mL, identifying 139 infected and 637 healthy quarters. The data were analyzed to test Random Forest Classifier(RFC), Logistic Regression Classifier, AdaBoost Classifier(ABC), and Naïve Bayes Classifier algorithms. The RFC models using milk features and temperature features achieved accuracy and precision scores of 0.88 and 0.88, respectively, yielding 0.90 AUC values, with high feature importance for milk lactose and SNF. Using only thermal imaging features, the ABC model yielded accuracy and precision of 0.82 and 0.67, respectively, with 0.56 AUC. We conclude that SCM prediction using machine learning algorithms is most promising when combined with thermal images and milk-based parameters. Future studies with larger datasets and refined methods for thermal image capture and analysis are warranted to validate these findings.
Lower urinary tract carcinomas in dogs are rare conditions with a poor prognosis. This prospective clinical study evaluated the safety, feasibility, and therapeutic efficacy of transarterial chemoembolization (TACE) using quick-soluble gelatin microparticles (QS-GMP) in 29 client-owned dogs with lower urinary tract carcinomas. Tumors primarily involved the bladder and urethra. Following selective catheterization of tumor-feeding vessels, intra-arterial carboplatin was administered prior to QS-GMP embolization. All patients underwent follow-up computed tomography (CT) scanning 4 weeks post-TACE. Tumor volume decreased in all cases, with a median reduction of 76.41% (95% confidence interval: 65.06-78.41%, p < 0.001). Partial remission was observed in 96.5% of patients; one case showed stable disease. Clinical signs, including pollakiuria, hematuria and stranguria, improved. No major complications occurred during the procedure or within two weeks after the procedure, and minor adverse effects were self-limiting. Repeat TACE was performed in 28/29 patients. The median survival time was 569 days for all patients and 534 days when non-tumor-related deaths were excluded. These findings suggest that TACE with QS-GMP may be an option for dogs with unresectable lower urinary tract carcinomas, with potential for repeated interventions and prolonged survival.
The architecture of bone defects presents a challenge in designing scaffolds that support structural integrity and enhance osteoinduction. To address this, a noninvasive injectable silk fibroin (SF)-based hydrogel incorporating magnetic nanohydroxyapatite (MHAp) and osteogenic growth peptide (OGP) was developed. This study investigated the osteogenic differentiation potential of the hydrogel and its underlying mechanism on canine periodontal ligament stem cells (cPDLSCs). The incorporation of MHAp and/or OGP significantly enhanced osteogenic differentiation as evidenced by increased alkaline phosphatase activity, osteogenic marker expression, and biomineralization. Proteomics analysis revealed that TGF-β, BMP, and Notch signaling pathways were involved in hydrogel-induced osteogenic differentiation, which was further validated by an inhibition assay. Overall, the injectable SF hydrogel containing MHAp and OGP demonstrates strong potential for clinical application in the treatment of alveolar bone defects.
Myxomatous mitral valve disease (MMVD) is leading cause of acquired heart failure in small-breed dogs. Although the American College of Veterinary Internal Medicine (ACVIM) staging is widely used in practice, how each stage and the degree of cardiac remodeling affect survival has not been clearly defined. We aimed to determine whether severe remodeling in dogs with preclinical (stage B2) disease is associated with survival differences compared with dogs with congestive heart failure (CHF; stage C) and less severe remodeling. This retrospective study analyzed 270 dogs with MMVD, stratified by the left atrium-to-aortic root ratio as having mild (<1.9), moderate (1.9-2.2), and severe (≥2.2) left atrial enlargement. Survival for dogs with stage B2 disease with severe LA enlargement did not differ significantly from dogs with stage C disease with mild-to-moderate enlargement. However, within the subgroup with severe LA enlargement, dogs that developed CHF had significantly shorter survival than those that remained in stage B2 (360 vs. 654 days; P = 0.032). Severe LA enlargement in dogs with preclinical disease contributes to a poor prognosis comparable to that of CHF with mild-to-moderate enlargement, indicating that stage alone may be insufficient for determining prognosis and that LA remodeling should also be considered.
The transition from gestation to lactation is a critical period that leads to insulin resistance (IR) in dairy cows. IR detection requires invasive sampling and laborious methods; therefore, novel and non-invasive approaches are essential. Extracellular vesicles (EVs) are nano-sized particles released by cells and are linked to disease pathophysiology. Hence, the present study aimed to explore the potential application of cow milk EVs (MEVs) in detecting IR in dairy cows. An intravenous glucose tolerance test (IVGTT) was performed (n = 12), and the cows were classified as low, moderate, or high IR based on prepartum area under the curve insulin values. MEVs were enriched using double-size exclusion chromatography and physico-chemically characterized. The results revealed that the particle diameter of MEVs was significantly higher (p ≤ 0.05) in the high-IR group compared to other groups. Out of 26 fatty acids (FAs) detected in MEVs, three long-chain FAs differed significantly in abundance (p ≤ 0.05) in high-IR group. Proteomic analysis revealed a correlation of inflammation and the IR-related proteins; Annexin A1 and Rab proteins, to IR severity. Overall, this study identified promising predictors of IR in cows, supporting the development of non-invasive biomarkers for early detection, facilitating improved herd and disease management, and reducing economic losses.
To investigate the influence of probiotic supplementation on parameters of renal function, dogs with chronic kidney disease (CKD) received a commercial probiotic formulation containing Enterococcus lactis SF68 (n = 8) or placebo (n = 8) for 60 days. Gut microbiome was investigated by comparing with healthy dogs (n = 10). Blood biochemistry, urinalysis, inflammatory and oxidative markers, uremic toxins and blood pressure were monitored. Higher presence of Lachnospiraceae family, Blautia bacterial species and Ruminococcus gnavus group was observed in dogs with CKD when compared with healthy dogs. Adding the probiotic to the diet decreased the abundance of Ruminococcus gnavus. Probiotic treatment resulted in a significant reduction in plasma concentrations of symmetric dimethylarginine (SDMA), from 1.50 ± 0.18 to 1.35 ± 0.16 µmol/l (p = 0.008), and indoxyl sulfate (IxS), from 19.1 ± 6.8 to 12.8 ± 4.8 µmol/l (p = 0.04). Cytokine inflammatory markers did not show significant changes. An increase in urine protein-to-creatine ratio, 1.5 ± 0.6 vs 1.2 ± 0.5, and in systolic blood pressure, 163 ± 11 vs 144 ± 6 mmHg (p = 0.033), was observed in the placebo group but not in dogs receiving probiotic. In conclusion, feeding Enterococcus lactis SF68 to dogs with CKD results in changes in intestinal microbiota that are associated with a decrease in plasma concentrations of IxS and SDMA and a reduction in proteinuria and systolic blood pressure.
Detailed analysis of stabled horse behaviour can reveal accurate information about its well-being. Advances in deep learning now allow these behaviours to be tracked without being invasive through the use of video data. This study evaluated a convolutional neural network for recognising standing, lying, and drinking behaviours in a horse housed in a wooden stall and recorded continuously over 29 consecutive days. Model predictions were compared with manually annotated ground truth data. Standing was detected with high precision (97.5%) and high recall (89.2%). Lying behaviour was classified with high precision (92.8%) but lower recall (63.1%). Activity patterns showed that standing dominated daily time budgets (>85%), lying accounted for 5-10%, and drinking occurred most often between 04:00 pm and 10:00 pm. These results demonstrate that deep learning can classify common equine behaviours from video, supporting its use in automated welfare monitoring. Future evaluations will explore the recognition of less frequent behaviours.
Staphylococcus aureus (S. aureus) is a Gram-positive bacterium that serves as a major pathogen causing bovine mastitis. One of the key intracellular survival mechanisms of S. aureus involves the induction of autophagy. The capacity of S. aureus to subvert autophagy for intracellular survival and replication is strongly linked to its broad array of virulence determinants, which play pivotal roles in infection establishment and immune evasion. One of these virulence factors is phenol-soluble modulin alpha (PSMα), an amphipathic small peptide that plays a key role in the pathogenesis of S. aureus. Utilizing gene-edited S. aureus strain and bovine mammary epithelial cells (BMECs, MAC-T), this study elucidates the involvement of PSMα in autophagy induction during S. aureus infection. The results demonstrated that PSMα knockout attenuated S. aureus-induced autophagy, weakened the activation of the PI3K/Akt/mTOR pathway, and reduced intracellular bacterial load. Our findings elucidate a PSMα-dependent autophagy induction mechanism in S. aureus infection, which is associated with the intracellular survival of S. aureus.
African swine fever (ASF) continues to expand worldwide. Recent detection in wild boar in Spain highlights the urgent need for effective control tools, with oral vaccination as a key priority. Following previous evaluation of the attenuated Lv17/WB/Rie1 strain, we assess an improved derivative, Lv17/WB/Rie1-ΔCD, lacking EP402R (CD2v) and EP153R, replaced by GFP to abrogate haemadsorption and enable Differentiating-Infected-from-Vaccinated-Animals (DIVA) diagnostics. Vaccinated animals received either a single high dose (104 TCID₅₀) or a prime and re-exposure regimen (102 TCID₅₀ plus a 104 TCID₅₀). Animals were challenged intramuscularly with the virulent Armenia07 genotype II strain. The ΔCD vaccine was well tolerated, inducing only transient low-grade fever. Prime-re-exposure vaccination induced earlier seroconversion (mean 12 ± 4 dpv) and sterilizing immunity in 5/6 animals in the high dose group. Overall protection reached 90%, while all unvaccinated controls died within 7 days. Quantitative PCR revealed >10³-fold reductions in viral genome copies in blood and tissues versus controls. DIVA ELISA reliably distinguished vaccine-induced antibodies from infection-derived responses. These findings identify Lv17/WB/Rie1-ΔCD as a safer oral ASFV vaccine candidate, addressing concerns raised with the parental Lv17/WB/Rie1 by increasing attenuation and supporting multi-gene deletion strategies. Further studies on safety, transmission, genetic stability, and environmental behaviour are required before large-scale field trials.
