Frontiers in Veterinary Science

BackgroundLymphedema, a chronic and incurable condition with limited therapeutic options, has limited options to quantitatively assess functional changes during its development; as a result, a deeper understanding of its pathophysiology remains hindered. To characterize lymphatic alterations and their association with disease pathology in a clinically relevant model in the rat, we developed a longitudinal iPhone-based volumetry method combined with non-invasive NIR analysis of lymphatic function. Material and methodsSecondary lymphedema was induced by surgery and single-dose irradiation. iPhone volumetry provided longitudinal measurements of hindlimb volume, while NIR imaging quantified the pumping function of major lymphatic vessels in the popliteal area. ResultsAmong 30 rats, lymphedema developed in 80%, defined as interlimb volume differences exceeding 5% and persisting through 14 days. In all rats with lymphedema, disease persisted until the end of the study at postoperative day 42 (P = 0.0015). NIR imaging revealed lymphatic dilation, dye extravasation, and lymphangiogenesis in affected limbs. Lymphatics in limbs with lymphedema exhibited increased contraction frequency, reduced amplitude, and diminished transport compared to baseline and contralateral controls (all P < 0.05). In contrast, rats that did no develop lymphedema showed no postoperative functional changes, although at baseline they displayed higher frequency and lower amplitude and transport compared with LE rats (all P < 0.001). Baseline transport values correlated negatively with swelling (r = -0.44, P = 0.002), as determined by ROC analysis, which yielded an AUC of 0.83, a sensitivity of 83.3%, and a specificity of 82.6%. Histopathology at day 42 confirmed significant dermal thickening and fat deposition in LE limbs (P < 0.001 and P = 0.002, respectively). ConclusionLongitudinal volumetry and NIR imaging applied to a clinically relevant animal model suggest a strong association between swelling and lymphatic function, which could provide deeper insight into lymphedema pathophysiology and represent valuable tools for future research and therapeutic development.

BackgroundWorking equids are fundamental to the socioeconomic structure of Mexicos small-scale agricultural sector, which accounts for 71.2% of the countrys active Agricultural Production Units (APUs). Despite their critical role in human rural livelihoods, food security, and sustainable development, these animals face systemic "statistical invisibility" within national and international productive frameworks. This study evaluates the long-term population dynamics and geographical distribution of working equids to analyze their current status amidst agricultural modernization. MethodsA retrospective analysis was conducted using national census data from 1970 to 2022 provided by the National Institute of Statistics, Geography, and Informatics (INEGI). Population trends for horses, donkeys, and mules were calculated using the Average Annual Variation Rate (AAVR). The severity of population declines was classified according to an adaptation of the International Union for Conservation of Nature (IUCN) criteria. Finally, national census records from INEGI, Agri-food and Fisheries Information Service (SIAP) and The Ministry of Agriculture and Rural Development (SADER) were contrasted with FAOSTAT database estimates to identify reporting discrepancies. ResultsBetween 1970 and 2022, the total equine population in Mexico decreased by 76.5%, falling from 6.8 to 1.6 million. However, a "paradox of modernization" was identified: while total numbers plummeted, the proportion of equids used specifically for work reached a historical peak of 81% in 2022, effectively having doubled from the 44% recorded in 2007. While donkeys and mules have suffered drastic total reductions (87% and 88%, respectively), working horses experienced a resilient 37% recovery between 2007 and 2022 (+3.71% AAVR). Furthermore, a staggering 710.8% discrepancy was found between national census data and FAOSTAT estimates, representing an overestimation of 11.3 million animals in international records. ConclusionsThe persistence and recent recovery of working equids reflect a "resilience of necessity" for approximately 500,000 APUs that depend exclusively on animal traction and packing due to economic constraints and complex topography. These findings challenge the narrative of total agricultural mechanization and highlight an urgent need for evidence-based public policies that address the statistical invisibility of working equids as indispensable drivers of rural sustainability and food security.

ABSTRACT Objective: This study aimed to systematically analyze the correlation between body weight and body measurement traits and the growth pattern of female Jiangyue donkeys. Methods: A total of 484 female Jiangyue donkeys were selected to determine their body weight (Y) and 11 body measurement traits, including withers height (X1), body length (X2), chest circumference (X3), cannon circumference (X4), head length (X5), neck length (X6), chest width (X7), chest depth (X8), rump height (X9), rump length (X10) and rump width (X11). SPSS 27.0 statistical software was used to conduct descriptive statistics, correlation and regression analysis on body weight and body measurement traits, and the optimal regression equation between them was established by the stepwise method and validated. Furthermore, four growth curve models (Logistic, Gompertz, Brody and Von Bertalanffy) were used to fit the body weight of 241 female Jiangyue donkeys at different months of age. Results: The body weight of female Jiangyue donkeys was significantly positively correlated with all body measurement traits (p<0.01), with the highest correlation coefficient observed for chest circumference. The optimal regression equations for body weight established by stepwise regression yielded R2 values of 0.918 and 0.844 for growing and adult donkeys, respectively (p<0.01). Among the four growth curve models, the Von Bertalanffy model exhibited the best fitting effect (R2=0.99992), with an estimated asymptotic body weight of 184.41 kg, which was close to the measured values of growing female donkeys. Conclusion: The regression models for body weight estimation and the Von Bertalanffy growth curve model for growth pattern evaluation established in this study can serve as effective tools and production management of female Jiangyue donkeys.

IntroductionAntimicrobial Resistance (AMR) presents a critical public health challenge, particularly in smallholder broiler farming, where antibiotics are often used preventively in the absence of effective biosecurity measures. ObjectiveThis study investigates the adoption of biosecurity practices as a sustainable alternative to antibiotics through Participatory Systems Mapping and Experimental Games. MethodsA participatory mixed-methods study was conducted in southern Bangladesh (September 2024-June 2025). Causal Loop Diagrams (CLDs) were co-created with farmers, dealers, and veterinary officers. Ten broiler farmers from single village were selected via purposive and snowball sampling. Experimental games simulated four production cycles where farmers chose Option A (biosecurity, adopters) or Option B (antibiotics, non-adopters) after several interactive trainings. Key metrics including biosecurity compliance (0-12 scale), mortality, FCR, antibiotic use, outbreak history, and economic outcomes were recorded. ResultsCLD analysis revealed a reinforcing loop of increased antibiotic reliance driven by fear of mortality, and balancing loops involving training, biosecurity practices, and consumer incentives to reduce use. Five farmers chose Option A, and both groups remained stable until Round 4. Adopters had flock sizes of 800-2000 birds (non-adopters, 600-1000; mean for both = 1000), were younger, and more educated compared to non-adopters. At baseline, both groups had similar biosecurity scores (0). Adopters had higher mean outbreaks (2 vs. 1.4), mortality (5.6 vs. 4.2), antibiotic use (3.6 vs. 3), and FCR (1.8 vs. 1.6) compared to non-adopters. By Round 4, adopters improved biosecurity scores by 125%, eliminated outbreaks, reduced mortality by 52.6%, stopped antibiotic use, improved FCR by 13.3%, and gained 71.72% profit per bird compared to non-adopters. Non-adopters, influenced by adopters, increased biosecurity scores by 25%, reducing outbreaks, mortality, antibiotic use, and FCR. Adopters also increased direct sales to consumers, yielding a 10%-16% profit gain per bird each round. ConclusionThis study highlights the successful adoption of biosecurity practices by farmers, replacing antibiotics and improving production outcomes. Farmer-driven adoption of these practices fosters long-term sustainability and supports a healthier planet within the One Health framework.

Restlessness is a symptom of chronic boredom in humans and a behavioural phenomenon anecdotally described as a concern in bulls raised for fattening purposes, but it has so far not been addressed in research. The two studies presented in this paper aimed to gain first insights into restlessness in bulls. We operationally defined restlessness by the number of transitions between behaviours in a given time period, and quantified restlessness in bulls of different weight classes (300, 400, 500 kg) on farms keeping bulls on fully-slatted floors (n=8, Study 1) as well as across three different husbandry systems (fully-slatted floor (FS, n=4), straw-based (SB, n=4) and organic pasture (OP, n=3), Study 2). All farms were visited twice, and the behaviour of different individuals was continuously recorded for 15 minutes each between 9 a.m. and 5 p.m. (Study 1) and for 8 minutes each between 6 a.m. and 10 p.m. (Study 2). The effects of weight class and husbandry system were analysed using generalised linear mixed-effects models, and we ran a sequence analysis to cluster observations by the sequence, frequency, and duration of bulls behaviours in Study 1. Bulls kept in fully-slatted floor systems in Study 1 changed their behaviour on average 48.3 times per 10 minutes, with high variability both within and across farms. Weight class did not have a statistically supported effect on the number of transitions, and the sequence analysis revealed four clusters that differed in sequence and in the number of transitions. In Study 2, OP bulls showed fewer transitions than SB and FS bulls (X22 = 23.6, p < 0.001), while SB and FS bulls did not differ. While SB pens were more structured and offered more space per animal, both SB and FS systems can be characterised by monotony, which may explain the similar level of restlessness in both systems. Alternatively, or in addition, the high feeding intensity in SB and FS systems may have caused the higher number of transitions compared to the OP system, potentially elicited by subacute ruminal acidosis and/or laminitis and the resulting pain. However, these explanations are speculative and require systematic disentanglement in future studies. This study provides initial insights into restlessness in bulls and lays the groundwork for future research to identify the causes underlying restlessness and investigate its association with bull welfare.

BackgroundThe gut-kidney axis plays a direct role in gastrointestinal and kidney health. Gut-derived metabolites like uremic toxins are associated with the pathophysiology of feline chronic kidney disease (CKD). The aim of the study was to identify novel fecal biomarkers and investigate the roles of gastrointestinal metabolites in feline CKD. ResultsFecal samples from 41 healthy non-CKD (control) and 67 CKD cats, including 5 IRIS stage 1 (CKD1), 37 stage 2a (CKD2a), 18 stage 2b (CKD2b), and 7 stage 3 (CKD3), were subject to fecal untargeted metabolomics and targeted short-chain fatty acid (SCFA) analyses. Multiple linear regression, adjusted for sex, age, body weight and study site, identified 64 differential metabolites between control and across CKD groups (P<0.0001 and FDR<0.10). Approximately 65% of the metabolites were lipids, including polyunsaturated long-chain fatty acids, acylcarnitines, and ceramides. Random Forest algorithm selected N1-methyl-2-pyridone-5-carboxamide (2PY), a uremic toxin from nicotinamide catabolism, as the top fecal marker for classifying feline CKD. Fecal 2PY was increased in CKD1 (P = 0.03), CKD2a, CKD2b, and CKD3 (all P<0.0001) compared to the controls. Data mining revealed serum concentration of 2PY was significantly increased with severity of CKD in cats, possibly due to impaired renal excretion. Cholesterol and arachidonic acid, markers for enterocyte shedding and inflammation, were increased in CKD3 versus control (both P<0.05). In healthy non-CKD cats, evident suggested fecal lipids increased with age (P<0.0001), and were higher in females versus males (P<0.0001). While fecal indole and p-cresol were increased in CKD3 versus control (both P<0.05), no change was observed in indoxyl sulfate (IS) or p-cresol sulfate (PCS). Fecal indole-3-acetic acid (IAA) was decreased in several CKD groups compared to the controls (all P<0.05). Finally, two branched SCFAs, isobutyrate and isovalerate, were increased in CKD3 versus control (both P<0.05). ConclusionsThe study revealed 2PY as a novel marker and unveiled profound alterations in intestinal lipid compositions with a potential link to gut barrier integrity and inflammation in CKD.

BackgroundPeriodontitis (Perio) is a progressive oral disease characterized by inflammation and degradation of the periodontal apparatus and is associated with local and systemic morbidity including loss of teeth, cardiovascular disease, and diabetes mellitus, among others. Perio is highly prevalent in domestic canines and exhibits certain parallels in pathogenesis and pathophysiology to Perio in humans, although standard treatments are less effective. In both species, a complex interplay between oral microbiota and host immune response is implicated in the etiology of Perio but is not fully understood. ResultsUsing shotgun metagenomics and RNA-seq on oral samples from companion dogs, we identify features of the oral microbiome and host transcriptional profile that are associated with Perio and its progression. We observe differences in microbiota composition between Perio and non-Perio animals that are largely consistent with what has been described in humans but also identify several species that are distinctly associated with canine Perio. We observe an abrupt shift in host gene expression related to immune response and tissue structure that is associated with disease severity, specifically the progression from mild periodontal disease (PD) to more severe Perio and the initiation of clinical attachment loss. The gingival plaque microbiota exhibits a parallel dynamic, with distinct compositional profiles in mild, moderate, and severe PD. We then examine several of the known mechanistic components of the keystone pathogen hypothesis of PD, identifying specific commonalities between canine and human pathologies, including the involvement of Porphyromonas species and related virulence factors. Additionally, we show infiltration of gingival tissue by Porphyromonas and Tannerella spp. via fluorescence microscopy. Finally, we assess correlations between host gene expression and microbial metabolic pathways which suggest additional potential virulence factors. ConclusionsThis work elucidates the metagenomic and transcriptomic signatures of Perio in companion dogs with the goals of informing veterinary medicine, evaluating the potential of canines as a model organism for the study of Perio, and clarifying the relationship between Perio development and progression, the oral microbiota, and the localized host response. Our findings provide insight into the etiopathogenesis of canine Perio and its relationship to human Perio and suggest novel targets of potential translational interest.

Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is an important zoonotic pathogen that disrupts intestinal epithelial barrier integrity and induces excessive inflammatory responses, thereby leading to impaired growth performance and intestinal injury. EHEC is also an important cause Hemolytic Uremic Syndrome (HUS) in children and older adults. In pig production, chitosan is considered a promising alternative to antibiotics due to its bioadhesive and antimicrobial properties, but the effects and underlying mechanisms of chitosan (COS) under pathogenic challenge remain to be elucidated. One hundred and eight pigs were randomly divided into three treatments: an unchallenged control group (CON), an EHEC-challenged control group (ECON), and an EHEC-challenged group supplemented with 100 mg/kg COS (ECOS). Results show that EHEC challenge increased the feed conversion ratio (FCR), increased inflammatory cytokine levels, disrupted intestinal morphology, and downregulated tight junction and nutrient transporter gene expression (P<0.05). Dietary COS supplementation significantly improved average daily gain (ADG) and FCR during day 6-14 (P<0.05). Moreover, COS reduced fecal shedding of total E. coli (P = 0.085) and EHEC, attenuated systemic inflammation by decreasing serum TNF- and IL-6 levels, and enhanced humoral immunity as indicated by increased IgA and IgM concentrations (P<0.05). Importantly, COS alleviated EHEC-induced intestinal injury by restoring villus height and villus-to-crypt ratio, with enhanced mucosal digestive enzyme activities, and upregulated expression of tight junction proteins (ZO-1 and occludin) and nutrient transporters (SGLT-1 and PEPT1) (P<0.05). In conclusion, these findings indicate that dietary COS improves growth performance in EHEC-challenged weaned pigs, with enhanced intestinal barrier integrity and nutrient transport capacity.

Equine asthma (EA) is a highly prevalent, chronic, inflammatory disease of the lower airways ranging from mild-to-moderate to severe clinical presentations. Diagnosis currently relies on bronchoalveolar lavage fluid (BALF) cytology, an invasive method associated with interobserver variability, which highlights the need for more reproducible approaches. MicroRNAs (miRNAs) are small noncoding RNAs involved in post-transcriptional gene regulation. They are stable and readily detectable in body fluids and have shown promising results as biomarkers in human asthma. The aim of this study was to characterize miRNA abundance profiles in BALF and serum from horses with distinct EA endotypes to evaluate their biomarker potential and explore their involvement in disease pathogenesis. A total of 43 horses were included and classified as either EA (n=32) or controls (n=11), based on clinical examination and BALF cytology. The EA horses were further divided into three endotypes based on BALF inflammatory cell composition: neutrophilic asthma (n=10), mastocytic asthma (n=15), and mixed asthma (n=7). RNA was isolated from both serum and BALF samples and analyzed by quantitative real-time PCR (qPCR) targeting 103 miRNAs linked to asthma and pulmonary inflammation in humans. Differential miRNA abundance was analyzed across EA endotypes. The most significantly differentially abundant miRNAs were used for in silico target prediction and pathway enrichment analyses. Horses with mixed EA had significantly lower levels of eca-miR-125a-3p and eca-miR-125b-5p in BALF compared to controls. Additionally, eca-miR-146a-5p abundance was significantly increased in BALF from horses with neutrophilic EA compared to mastocytic EA. Target and pathway enrichment analyses for eca-miR-146a-5p identified immune-relevant pathways, such as MAPK and T-cell receptor signaling, supporting its involvement in inflammatory processes associated with asthma. This study identified three promising candidates, eca-miR-125a-3p, eca-miR-125b-5p, and eca-miR-146a-5p, as potential biomarkers associated with different EA endotypes. These miRNAs are interesting candidates for further investigation in an independent cohort.

ObjectiveStall housing of pregnant sows raises welfare concerns, whereas conventional group housing systems often reduce space efficiency. This study evaluated the effects of periodic group housing (PG) on reproductive performance and welfare compared with continuous stall housing (CS). MethodsSows in the CS group (n = 15) were continuously housed in stalls. In the PG group (n = 15), sows were housed in groups of three and allocated 1 day of group housing and 6 days of stall housing per week over 10 weeks. During group housing sessions, the sows had access to a group housing area containing sawdust. Behavioral observations and salivary cortisol measurements were conducted on the first day of the stall housing session in weeks 1, 6, and 10. Behavioral indices were expressed as proportions based on 90 sampling points recorded at 1-min intervals. ResultsThe number of stillbirths was significantly lower in the PG group than in the CS group (0.63 vs. 1.49 piglets per litter). whereas other reproductive outcomes, including total litter size and average birth weight, did not differ. In older parity sows, the PG treatment markedly increased the proportion of time spent lying, suggesting reduced discomfort associated with restricted movement. Furthermore, the proportion of exploratory behavior decreased markedly, and drinking behavior showed a decreasing trend across parity levels in the PG group, suggesting partial satisfaction of motivations for environmental exploration and oral manipulation. The proportion of oral abnormal behavior showed a pronounced interaction between housing treatment and experimental week, increasing from week 1 to week 6 in the PG group. Salivary cortisol concentrations did not differ between the groups. ConclusionPG may improve reproductive performance and partially satisfy the behavioral motivations restricted under continuous stall housing. This system may represent a practical alternative for improving animal welfare while minimizing economic losses.

Skeletal muscle regeneration is often impaired after acute muscle damage induced by viperid snake venoms, such as that of Bothrops asper, a medically-relevant species in Latin America. It has been shown that traces of venom that remain in the damaged muscle affect myogenic cells in culture, raising the possibility of inhibition of these toxins during the regenerative process as a way to improve regeneration. Using a mouse model of myonecrosis and regeneration, we evaluated the effects of Varespladib (a phospholipase A2 inhibitor) or Marimastat (a metalloproteinase inhibitor) on muscle regeneration when administered intravenously 24 h after the onset of myonecrosis, i.e., after muscle damage has occurred. The regenerative process was evaluated 14 and 28 days after venom injection. Results show that Marimastat, or a combination of both inhibitors, improved the extent of skeletal muscle regeneration and reduced the extent of tissue fibrosis when compared to tissue from mice receiving venom and no inhibitors, as judged by qualitative and quantitative histological assessment. Results underscore the deleterious role of traces of venom components in the damaged muscle during muscle regeneration and suggest that the administration of metalloproteinase inhibitors, or a combination of metalloproteinase and phospholipase A2 inhibitors, even when muscle damage has developed, may be a therapeutic alternative for improving the extent of muscle regeneration.

Cetacean pathology is a cornerstone for population and marine ecosystem health monitoring, allowing clear differentiation among natural and anthropogenic threats. Previous studies in the Canary Islands reported natural causes of death in 59.4% (1999-2005) and 81% (2006-2012) of stranded cetaceans, versus anthropogenic causes in 33.3% and 19%, respectively. This study aimed to determine the causes of death (CD), pathologic findings, and epidemiological patterns of 316 cetaceans stranded in the Canary Islands between 2013 and 2018. The CDs were classified in pathologic entities (PEs) emphasizing natural versus anthropic origins. Of 316 animals, 224 (70.9%) from 18 species were suitable for pathological investigations. Among natural PEE, natural pathology associated with good nutritional status (NP-GNS) and natural pathology associated with significant loss of nutritional status (NP-LNS) represented 43/224 (19.2%) and 36/224 (16%) cases, respectively. Natural pathology with undetermined nutritional status (NP-UNS) occurred in 19/224 (8.5%) animals. Intra- and interspecific traumatic interactions (ITI) represented 30/224 (13.4%) cases, followed by neonatal/perinatal pathology (NPN) 19/224 (8.5%) and live-stranding stress and/or capture myopathy (LS-CM) 18/224 (8%). Infectious and parasitic diseases predominated in natural PEs. Anthropogenic PEs included interaction with fishing activities (IFA) in 17/224 (7.6%) cases, vessel collisions (VC) in 9/22 (4%) cases, and foreign body-associated pathology (FBAP) in 3/224 (1.3%) animals. Overall, anthropogenic causes accounted for 12.9% of deaths, natural causes for 73.6%, and the CD could not be established in 30/194 (13.4%) cases. This study reaffirms the trends concerning recognized PEs (NP-GNS, NP-LNS, NP-UNS, ITI, NPN, LS-CM, IFA, VC, and FBAP), expands the body of knowledge on cetacean pathology in the Canary Islands, and reports novel findings including mixed infections, clostridiosis in uncommon species, uremic syndrome secondary to urethral nematodiasis, gas embolism in unusual species, epibiont stomatitis, congenital musculo-skeletal malformations, or neoplastic processes. These findings advance understanding of cetacean mortality patterns and support conservation and management strategies.

Background and aimTrauma healing with low-intensity ultrasound is effective for different types of injuries affecting both soft tissues and bones. The work aimed to disclose the healing potential of a new type of ultrasound, ultra-wideband low-intensity mechanical waves (UMUS), and to compare its effects with those of low-intensity pulsed ultrasound (LIPUS) in a model of trauma. Material and methodsThe work was performed on 2-to 3-month-old male Wistar rats. The model of tail amputation was created, and a transducer emitting UMUS (1-7 MHz, 0.22 mW/cm2) was applied daily for 10 days to the surface of the trauma site in animals that were timely immobilized. LIPUS (1.5 mHz, 30.0 mW/cm2) was used in a separate group of animals. Sham-stimulated rats were used as a control. The intensity of collagen expression in the subdermal tissue was assessed in van Gieson-stained sections, whereas in the UMUS group, expression of CD31, CD34, VEGF, and Ki67 was analyzed. ResultsStarting on the 20th day after trauma, UMUS-treated animals demonstrated a statistically significant decrease in the surface area of the traumatic zone compared to the control, whereas LIPUS-treated rats showed this difference on the 30th day of observation. Starting from the 30th day, a significantly greater reduction in the surface of trauma was observed in UMUS, with complete closure achieved in 6 out of 9 rats (P=0.019 vs control), whereas in LIPUS-treated animals, a similar result was observed in 2 out of 8 rats (P>0.05). In UMUS-treated rats, heightened expression of collagen in animals with LIPUS exceeded control data by 7.84% (P=0.034), while the expression in rats with UMUS exceeded data in LIPUS-treated rats by 14.71% (P=0.013). Increased expression of CD31, CD34, VEGF, and Ki67 was observed in UMUS-treated rats. ConclusionsUMUS treatment accelerated healing and reduced wound size, and increased the expression of collagen, CD31, VEGF, CD34, and Ki67, supporting angiogenesis and collagen formation. Effects are more pronounced compared to LIPUS treatment. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=176 HEIGHT=200 SRC="FIGDIR/small/717366v1_ufig1.gif" ALT="Figure 1"> View larger version (75K): org.highwire.dtl.DTLVardef@936775org.highwire.dtl.DTLVardef@16d505eorg.highwire.dtl.DTLVardef@1b75d7dorg.highwire.dtl.DTLVardef@15ae99c_HPS_FORMAT_FIGEXP M_FIG C_FIG

The early evolutionary history of modern domestic horses (Equus caballus/E. ferus caballus), known as the DOM2 lineage, is well documented due to numerous archaeological and ancient DNA (aDNA) studies. Although many uncertainties remain in the domestication timeline, current evidence suggests that the domestication of modern horses began in the Pontic-Caspian steppe at least [~]2700 BCE (before common era), or even earlier. However, it is not known how long remnant wild horse populations survived or when domestic horses were introduced into Northern Europe. In this study, we review the current knowledge of horse domestication, focusing on Northern Europe. We analysed prehistoric horses from western Russia to assess the body sizes of wild horses from the Ivanovskaya site (5900-3800 BCE) in the Pontic-Caspian steppe, and the body weight of one Lithuanian wild horse (4000-3800 BCE). Additionally, we analysed body sizes of Late Bronze Age-Early Roman Age horses (1100 BCE-300 CE; common era) and re-analysed body sizes and estimated rider weights of historic domestic horses from Lithuania (100-1400 CE). We searched for pathological changes and signs of bit wear indicative of bridling. Furthermore, we investigated maternal genetic diversity by sequencing ancient mitochondrial DNA. We found that wild horses from Ivanovskaya were intermediate in body size between earlier and more recent horses of the Eurasian Steppe, and that the Lithuanian wild horse weighed only [~]270 kg and Late Bronze Age-Early Roman Age horses 200-300 kg. Lithuanian domestic horses were pony-sized (< 130 cm on average). Bit wear was confirmed on one tooth, the oldest domestic horse in Lithuania (799-570 cal BCE). Another tooth showed signs of the Equine Odontoclastic Tooth Resorption and Hypercementosis (EOTRH) condition. Mitochondrial DNA was successfully amplified from one Ivanovskaya wild horse along with 25 other ancient samples, including Lithuanias oldest domestic horse. mtDNA diversity was high, revealing several maternal lineages.

Compassion fatigue is a well-documented hazard among healthcare and veterinary professionals, yet the psychological toll on informal caregivers of feral cat colonies, likely numbering several tens of thousands in Portugal, remains largely unexplored. This cross-sectional study examines internal and external factors associated with the secondary traumatic stress component of compassion fatigue among 172 informal caregivers in Portugal. Secondary traumatic stress refers to work-related secondary exposure to individuals who have experienced extremely stressful or traumatic events. Structured telephone interviews assessed sociodemographics, colony management, compassion satisfaction, resilience, spiritual well-being, and perceived social support. Univariate and multivariable linear regression identified predictors of secondary traumatic stress. Results indicate that 47% of participants experienced moderate secondary traumatic stress, and 10% reported high levels. Multivariable analysis revealed that caring for large colonies (more than 25 cats) and being unemployed were significantly associated with higher fatigue. Conversely, older age, higher perceived family support, and the resilience dimension of serenity served as protective factors. Interestingly, finding meaning in life was positively correlated with fatigue, suggesting that caregivers who perceive their role as central to their life purpose may become more emotionally invested, increasing vulnerability to distress when unable to help animals. Official colony registration and formal institutional support did not significantly alleviate fatigue. These findings highlight that institutional support alone is insufficient to mitigate fatigue among informal caregivers, who experience significant distress driven by both practical burdens and profound emotional involvement. The most frequently reported concern among caregivers was the inability to cover the costs of feeding and veterinary care for the cats. Interventions must address both external needs (e.g., support to cover veterinary and feeding expenses for the cats) and internal coping mechanisms. Implementing psychosocial support alongside trap-neuter-return programs may also improve caregiver well-being and foster sustainable urban feral cat management. This underscores a One Health perspective, demonstrating that animal health is closely interconnected with human well-being and environmental health.

ABSTRACT Background Dog bite injuries are a major yet largely preventable public health concern worldwide. They contribute significantly to morbidity, healthcare burden, and economic costs, particularly in emergency department .The present study aims to analyse the demographics and injury pattern of dog bite cases presenting to the emergency department of a tertiary care hospital in Chennai. Methods We conducted a cross-sectional study with dog bite injured participants attending the Causality from November 2025 to April 2026 data was collected using a structured tool including details on demographics (Age, Gender, Education) injury related characteristics , history of pure bite site of dog bite injury type, WHO bitten criteria and information to management etc. We used here non probability statistical analysis and age specific dog bite cases and independent variables were analysed using SPSS (2.0 version). Result A total of Two hundred sixteen dog bite cases were analysed in the study by period of 6 months The majority of participants were 172 (79.6%) were above 18 years and 44 (20.4) were below 18 year, 130 (60%) from rural areas and 86 (39.8%) from urban areas, 136 (63.0%) of Victims presented within a day of the bite, 61( 28.2) next day and 19 (8.8%) in after one week 66 (30.6) were bitten by own dog and 150(69.4%)were bitten by neighbour / friended dog. 124(57.4) were bitten by stray dog 92(42.6) bitten by pet dog. 117(54.2) were vaccinated dog and 99(45.8%) were not vaccinated .110(50.9) victims were injured by laceration. 26(12.0%) were injured by puncture wound.46(21.3) were injured by abrasion 10(4.6) were injured by avulsion 15(6.9%) were injured by crush injury. 156(72.2%) were had minor wound.45(20.8%) victims had moderate wound and 15(6.9%) victims had severe wound. 112(51.9%) victims were taken antibiotics.104(48%) were not taken antibiotics. 185(85.6%) victims received tetanus toxoid, 31(14.4%) were not received tetanus toxoid. CONCLUSION There is a high burden of dog bite injuries from stray dogs in India. Despite early hospital presentation in many cases gaps in first aid practices and rabies post exposure prophylaxis were evident and highlighting inadequate awareness. Key words Rabies immunoglobulin, Dog bite, WHO criteria, Anti rabies vaccine, stray Dog, wound

Disruptions within the microbiome-gut-bone axis are increasingly recognized as key contributors to impaired bone metabolism and leg disorders in broiler chickens. This study investigated the effects of a combined dietary additive containing Bacillus subtilis DSM 29784 and a phytogenic blend of garlic and essential oil components (BsP) on the modulation of microbial communities, intestinal integrity, mineral utilization, and bone-associated immune-osteogenic pathways. Five hundred and sixty-one-day-old male Ross 308 broilers were randomly assigned to a basal control diet or the same diet supplemented with BsP for 42 days, with eight replicates per treatment. Growth performance, cecal microbiome composition, jejunal tight junction expression, pro-inflammatory cytokines, ileal calcium-phosphate transporters, and femoral inflammatory and osteogenic gene expression were evaluated. The results demonstrated that BsP supplementation significantly improved body weight, weight gain, and feed conversion ratio while enhancing intestinal barrier function. Birds receiving BsP displayed upregulated expression of tight junction-related genes (CLDN-1, OCLD-1, TJP-1, MUC-2) and reduced jejunal inflammatory markers (TNF-, NF-{kappa}B). Improved mineral transport capacity was indicated by increased ileal CaSR and NaPi-IIb expression. Microbiome profiling revealed higher species richness (Chao1 and Shannon indices; P<0.05) and diversity (Bray-Curtis, PERMANOVA; P <0.001) on days 21, 35, and 42, with enrichment of beneficial taxa such as Clostridium butyricum, Enterococcus faecium, Lactobacillus salivarius, L. crispatus, and Bifidobacterium longum, accompanied by reduced Escherichia coli, and Enterococcus cecorum. Functional predictions suggested activation of serotonin-, melatonin-, and L-tryptophan-related pathways, indicating engagement of the microbiome-gut-brain axis. At the skeletal level, BsP reduced femoral expression of IL-6, IL-17, TNF-, and NLRP3 and enhanced BMP-2, SMAD-1, RUNX-2, and SPARC, aligning with improved mineral deposition. Network analysis revealed distinct inflammation-, bone-, and microbiota-dominant modules, highlighting the structured interactions linking microbial signals to osteoimmunological responses. Overall, BsP effectively modulated the microbiome-gut-bone axis, supporting intestinal homeostasis, mineral absorption, and bone formation. These findings underscore the potential of BsP as a functional feed additive to promote both intestinal and skeletal health in broilers.

Giraffes, unlike other large mammals, typically pose minimal risk to humans, their animals, and crops, so they are traditionally not involved in human-wildlife conflict. Tree crops, however, are expanding across Africa, resulting in crop raiding by giraffes and retaliatory snaring, poaching, and killing of giraffes in response. The dynamics of giraffe crop raiding, however, are poorly understood, making effective interventions difficult to implement. To better understand key factors for humans and giraffes that mediate crop raiding, we used a multi-method approach to estimate giraffe abundance and activity, understand farmers perceptions and decisions, and test countermeasures around Garissa Giraffe Sanctuary in eastern Kenya. We hypothesized that 1) giraffe farm invasion would occur in dry seasons, 2) farms growing mangoes would be more likely to be invaded, 3) reducing invasion with only physical barriers would be less effective than adding behavior-based countermeasures, 4) perceptions would match giraffe activity and 5) countermeasure adoption would be driven by cost. We found that invasion and crop raiding primarily occur during the dry season and are associated with mangoes. Farmers are using many countermeasures. Effective countermeasures target giraffe behavior combined with physical barriers. Countermeasures are most effective when negative associations with humans are reinforced. Floodlights and speakers that play predator calls both reduce invasion, but only if used consistently. Overall, farmers perceptions matched giraffe dynamics. Availability was the most important factor in farmers willingness to try a countermeasure. Our results suggest that conflict can be reduced and there is interest from farmers in doing so, but use of countermeasures should be consistently applied and supported by making necessary equipment and instructions available.

ObjectiveAnterior cruciate ligament (ACL) tears increase the risk for developing posttraumatic osteoarthritis (PTOA). Females have greater risk for both. However, studies defining sex-specific protein responses in human cartilage after ACL injury are lacking. We hypothesize that articular cartilages response to an injurious environment differs depending on sex. DesignWe compared the proteomic profiles of normal cartilage with injured cartilage harvested from the intercondylar area during ACL surgery. Sex-specific injury effects were estimated through contrasts between Injured Male and Normal Male and between Injured Female and Normal Female. Pathway enrichment analysis was done using gene ontology (GO) and compared against the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Extracellular matrix (ECM) proteins were further analyzed using the Matrisome AnalyzeR. ResultsFrom the 2,188 proteins identified, males and females shared 1,121 upregulated and 23 downregulated proteins in injured compared to normal cartilage. Analysis of ECM proteins and enriched pathways revealed mostly similar male and female responses to an injurious environment, with evidence of early cartilage remodeling in both sexes. Nevertheless, more than 240 proteins were affected specifically by sex, and significant sex differences were found in inflammation, ECM-related, and metabolic pathways. Males were enriched mostly in "ECM-receptor interaction", while females were enriched in "Citrate cycle (TCA cycle)", "Fatty acid degradation", and "Fatty acid metabolism" pathways. ConclusionArticular cartilage shows signs of remodeling soon after ACL injury, even when only exposed to an injurious environment rather than being physically impacted. Sex differences were observed in inflammation, metabolic pathways, and ECM synthesis.

Boar semen contains spermatozoa and seminal plasma (SP) that carries extracellular vesicles (EVs) among other components. However, artificial insemination (AI) doses produced by AI companies are highly diluted based solely on sperm concentration. The aim of this study was to evaluate the integrity of EVs isolated from AI doses, characterize the protein and miRNA content from high-fertility (HF) and reduced-fertility (RF) boars, and evaluate their functional impact on spermatozoa after dilution by a coincubation up to 24 hours at 38 {degrees}C. Proteomics identified 108 differentially expressed proteins between HF and RF EVs (97 upregulated in HF, 11 in RF), and transcriptomics revealed 80 differentially expressed miRNAs (DEMs) in EVs, 52 in SP, and 3 in spermatozoa, showing inverse expression in various shared DEMs between fertility rates, suggesting compartment-specific regulation. Functional coincubation demonstrated that EVs remain biologically active after dilution. HF EVs improved sperm quality parameters and reduced oxidative stress, while RF EVs increased total and progressive motility. Overall, our findings show that EVs from AI doses retain structural integrity, carry fertility-associated protein and miRNA signatures, and functionally modulate sperm quality in vitro. These features highlight porcine EVs as promising biomarkers and potential tools to optimize reproductive performance in swine production.