Arterial Blood Supply of the Stifle Joint in Horses.

The vascularization pattern of the equine stifle joint is insufficiently described in the literature, even though there is a growing need for knowledge of the exact blood supply, as (i) arthroscopy and endoscopic surgery techniques are increasingly performed in horses and (ii) ex vivo models of menisci need nutrient supply that mimic the in vivo situation. The aim of this study was to describe the vessels involved in the stifle joint supply and the exact branching pattern of the popliteal artery. Colored latex was injected into the arteries of nine pelvic limbs of equine cadavers (n = 6) to evaluate the occurrences, variations and approximate diameters of vessels that supplied the stifle joints. Next to a branch of the saphenous and descending genicular arteries, eleven branches of the popliteal artery could be described in horses that feed the vascular network of the stifle joint. With a focus on the blood supply of the menisci, a vascularization map was created to show the main influx to these intra-articular structures in detail. These findings are potentially of great importance to both clinicians in preparation of best-suited incisions for arthroscopy and researchers designing new approaches for meniscal studies and choosing suitable animal models.

Assessment of a Teaching Module for Cardiac Auscultation of Horses by Veterinary Students.

Auscultation of heart sounds is an important veterinary skill requiring an understanding of anatomy, physiology, pathophysiology and pattern recognition. This cross-sectional study was developed to evaluate a targeted, audio-visual training resource for veterinary students to improve their understanding and auscultation of common heart conditions in horses. Fourth- and fifth-year 2021 and 2022 Bachelor of Veterinary Science students at the University of Queensland (UQ) were provided the learning resource and surveyed via online pre- and post-intervention surveys. Results were quantitatively analyzed using descriptive statistics and Mann-Whitney U tests. Open-ended survey questions were qualitatively analyzed by thematic analysis and Leximancer™ Version 4 program software analysis. Over the two-year period, 231 fourth-year and 222 fifth-year veterinary students had access to the resource; 89 completed the pre-intervention survey and 57 completed the post-intervention survey. Quantitative results showed the resource helped students prepare for practicals and their perception of competency and confidence when auscultating equine cardiac sounds improved (p < 0.05). Compared to fifth-year students, fourth-year students felt less competent at identifying murmurs and arrythmias prior to accessing the learning resource (p < 0.05). Fourth-year and fifth-year students' familiarity with detection of murmurs improved after completing the learning resource (p < 0.001). Qualitative analysis demonstrated a limited number of opportunities to practice equine cardiac auscultation throughout the veterinary degree, especially during the COVID-19 pandemic, and that integrated audio-visual resources are an effective means of teaching auscultation.

The Detection of Thoracolumbar Spine Injuries in Horses with Chronic Laminitis Using a Novel Clinical-Assessment Protocol and Ultrasonographic Examination.

Postural adaptation is a prominent feature in horses affected by laminitis. Laminitis induces intense pain, especially in the forelimbs, prompting affected horses to assume a caudally displaced trunk posture, resulting in the hyperflexion of the thoracolumbar spine. This study assessed the nature and prevalence of thoracolumbar injuries in horses with chronic laminitis compared to horses without it. Sixty horses were used (thirty laminitic and thirty non-laminitic) of different athletic purposes and ages (2-20 years). The experimental protocol entailed a single assessment of horses' thoracolumbar spines, utilizing physical examination by MACCTORE, a scoring system developed specifically for this study. Additional evaluations included the Grimace Equine Pain Scale (HGS) and ultrasound exams. Statistical tests were used to compare values (Mann-Whitney or t-test) and lesions prevalences (Fisher) between groups (p < 0.05). The results showed a higher pain manifestation (HGS and heart rate, p < 0.0001) and thoracolumbar-spine-injury levels in chronic laminitis horses, both in MACCTORE clinical examinations (11.7 ± 4.8 vs. 4.2 ± 3.3, p < 0.0001) and general ultrasonographic indices (39.6 ± 12.0 vs. 20.7 ± 7.1, p < 0.0001), including specific examination approaches for various spinal elements. Horses with laminitis presented with a 14-fold higher prevalence of ultrasound-relevant lesions in the thoracolumbar spine (CI: 4.4 to 50.6, p < 0.0001) compared to controls. These findings constitute new evidence of an association between chronic laminitis and the presence of thoracolumbar spine injuries in horses, which may be confirmed by more sophisticated study designs.

Neonatal and maternal upregulation of antileukoproteinase in horses.

Introduction: The end of gestation, ensuing parturition, and the neonatal period represent highly dynamic phases for immunological changes in both mother and offspring. The regulation of innate immune cells at the maternal-fetal interface during late term pregnancy, after birth, and during microbial colonization of the neonatal gut and other mucosal surfaces, is crucial for controlling inflammation and maintaining homeostasis. Innate immune cells and mucosal epithelial cells express antileukoproteinase (SLPI), which has anti-inflammatory and anti-protease activity that can regulate cellular activation. Methods: Here, we developed and validated new monoclonal antibodies (mAbs) to characterize SLPI for the first time in horses. Peripheral blood and mucosal samples were collected from healthy adults horses and a cohort of mares and their foals directly following parturition to assess this crucial stage. Results: First, we defined the cell types producing SLPI in peripheral blood by flow cytometry, highlighting the neutrophils and a subset of the CD14+ monocytes as SLPI secreting immune cells. A fluorescent bead-based assay was developed with the new SLPI mAbs and used to establish baseline concentrations for secreted SLPI in serum and secretion samples from mucosal surfaces, including saliva, nasal secretion, colostrum, and milk. This demonstrated constitutive secretion of SLPI in a variety of equine tissues, including high colostrum concentrations. Using immunofluorescence, we identified production of SLPI in mucosal tissue. Finally, longitudinal sampling of clinically healthy mares and foals allowed monitoring of serum SLPI concentrations. In neonates and postpartum mares, SLPI peaked on the day of parturition, with mares returning to the adult normal within a week and foals maintaining significantly higher SLPI secretion until three months of age. Conclusion: This demonstrated a physiological systemic change in SLPI in both mares and their foals, particularly at the time around birth, likely contributing to the regulation of innate immune responses during this critical period.

Impact of platelet lysate on immunoregulatory characteristics of equine mesenchymal stromal cells.

Multipotent mesenchymal stromal cells (MSC) play an increasing role in the treatment of immune-mediated diseases and inflammatory processes. They regulate immune cells via cell-cell contacts and by secreting various anti-inflammatory molecules but are in turn influenced by many factors such as cytokines. For MSC culture, platelet lysate (PL), which contains a variety of cytokines, is a promising alternative to fetal bovine serum (FBS). We aimed to analyze if PL with its cytokines improves MSC immunoregulatory characteristics, with the perspective that PL could be useful for priming the MSC prior to therapeutic application. MSC, activated peripheral blood mononuclear cells (PBMC) and indirect co-cultures of both were cultivated in media supplemented with either PL, FBS, FBS+INF-γ or FBS+IL-10. After incubation, cytokine concentrations were measured in supernatants and control media. MSC were analyzed regarding their expression of immunoregulatory genes and PBMC regarding their proliferation and percentage of FoxP3+ cells. Cytokines, particularly IFN-γ and IL-10, remained at high levels in PL control medium without cells but decreased in cytokine-supplemented control FBS media without cells during incubation. PBMC released IFN-γ and IL-10 in various culture conditions. MSC alone only released IFN-γ and overall, cytokine levels in media were lowest when MSC were cultured alone. Stimulation of MSC either by PBMC or by PL resulted in an altered expression of immunoregulatory genes. In co-culture with PBMC, the MSC gene expression of COX2, TNFAIP6, IDO1, CXCR4 and MHC2 was upregulated and VCAM1 was downregulated. In the presence of PL, COX2, TNFAIP6, VCAM1, CXCR4 and HIF1A were upregulated. Functionally, while no consistent changes were found regarding the percentage of FoxP3+ cells, MSC decreased PBMC proliferation in all media, with the strongest effect in FBS media supplemented with IL-10 or IFN-γ. This study provides further evidence that PL supports MSC functionality, including their immunoregulatory mechanisms. The results justify to investigate functional effects of MSC cultured in PL-supplemented medium on different types of immune cells in more detail.

Pressure pain mapping of equine distal joints: feasibility and reliability.

Background: Osteoarthritis is a prevalent degenerative joint disease initiating chronic pain and lameness in horses. While several objective gait analysis systems have been developed and validated to quantify lameness severity in horses, methods to evaluate whether peripheral sensitization contributes to the pain experienced are missing. Objectives: To evaluate whether periarticular pressure pain mapping could be proposed as an auxiliary assessment tool in horses. Specific aims were to evaluate the feasibility and intra- and inter-rater reliability of pressure pain thresholds (PPT) determination at sites overlying the distal thoracic limb joints of clinically healthy horses. Study design: Prospective, randomized validation study. Methods: For feasibility assessment, PPT were measured with a hand-held digital algometer at six periarticular landmarks (2 sites per joint, 3 joints) bilaterally on the distal thoracic limb of 40 healthy horses (20 warmblood and 20 Freiberger). The joints tested were the metacarpophalangeal, on the latero-palmar and dorsal aspects (L-MCP and D-MCP), the proximal interphalangeal, on the dorsal and palmar aspect (D-PIP and P-PIP) and the distal interphalangeal, on the dorsal and lateral aspect (D-DIP and L-DIP). A feasibility score, ranging from 0 to 5, was attributed to each testing session. For intra- and inter-rater reliability assessment, L-MCP and D-MCP were selected to be tested again at 2 weeks intervals in 20 out of the 40 horses. Data were analyzed using a mixed-effect linear model to test differences in threshold per site and limb. Intra- and inter-rater correlation was calculated. Bland-Altman plots were performed to evaluate the variability of the measures. Results: The procedure was considered feasible (score <2) in 95% of horses (95% CI 88%-100%). Overall, median [interquartile range (IQR)] PPT was 9.4 (7.5-11.3) N. No significant side differences were found. P-PIP and D-DIP recorded significantly lower PPT (p < 0.001 and p = 0.002, respectively) than L-MCP. Median (IQR) were 9.9 (7.3-12.4) N, 8.4 (6.1-10.5) N and 9.0 (7.4-10.6) N for L-MCP, P-PIP and D-DIP, respectively. The intra-rater agreement was 0.68 (95% CI 0.35-0.86) for L-MCP, and 0.50 (95% CI 0.08-0.76) for D-MCP. Inter-rater agreement was 0.85 (95% CI 0.66-0.94) for L-MCP and 0.81 (0.57, 0.92) for D-MCP. Main limitations: Evaluation of feasibility was performed only for distal thoracic limbs joints; no data are provided for hind limbs or proximal joints. Only warmblood and Freiberger horses were included. Intra- and inter-rater reliability assessments were performed exclusively on data collected at the MCP joint. Conclusion: Pressure pain mapping of distal thoracic limb joints was feasible in horses. Local sensitivity differed among sites and no side differences were noticed. Data collected from the MCP joint suggest highly variable, subject dependent intra-rater reliability, ranging from poor to good, and good to excellent inter-rater reliability. Further studies evaluating pathologic vs. healthy joints are needed before recommendations can be made about clinical usability and diagnostic validity.

The use of genomic assisted selection could allow early selection among Thoroughbreds and Polo Argentino horses for practicing polo.

The Polo Argentino (PA) horse is a recognized breed, developed originally by mixing crossbred and Thoroughbred (TB) horses to play polo. Early PA selection is difficult due to unreliable performance estimations. This study investigated the usefulness of genomic markers previously linked to morphological and functional traits as a tool for the early selection of PA. To this, we genotyped 520 PA and 30 TB horses using the Equine GGPArray (Illumina, n=71,778 SNPs). Analyses included a genetic characterization of six genetic markers associated with behavioral (DRD4), muscular development (MSTN), and body size (LCORL, HMGA6, ZFAT, and LASP1) genes. Genetic differences in the DRD4, MSTN, and LCORL SNP were found between the two breeds, in the last two FST index between breeds was 0.13 and 0.6, respectively (P<0.01). In DRD4, G allele was the more prevalent in PA (0.56 vs 0.45 in TB, P<0.05), but no differences were observed between the genotypes associated with phenotypes. In MSTN, heterozygous genotypes were the most common in PA (48%), with a significant decrease in AA (Hardy-Weinberg P<0.05), suggesting a negative selection against it in polo horses. In body size, HMGA2 was monomorphic in all horses, while ZFAT and LASP1 SNP showed higher variability. Interestingly, 99% of PA showed a TT genotype in LCORL (only 66% in TB), demonstrating selection for smaller horses. Our results suggest that empirical selection in PA has generated an incipient genomic differentiation in discrete traits which could be used as a marker-assisted selection tool for early selection of polo horses.

Neurosurgical injuries associated with recreational horseback riding: a 6-year retrospective analysis.

INTRODUCTION: Horseback riding can cause severe brain and spinal injuries. This study aimed to identify the spectrum of neurosurgical injuries related to recreational horseback riding. METHODS: A retrospective study was performed utilizing the University of Puerto Rico neurosurgery database to identify patients who were consulted to the neurosurgery service between 2018 and 2023 after a horse fall during recreational activities. The outcome upon discharge using the modified Rankin scale (mRS) was documented. Descriptive statistics were used to report frequency and median values. RESULTS: The neurosurgery service evaluated and managed 112 patients with a horseback riding fall-related injury during six years. Ninety-eight (87.5%) patients were male. The patients' median age was 31.5 (IQR 22-40). There were 89 head injuries (79.5%), 19 spinal injuries(17%), and four combined head/spine injuries (3.5%). Forty percent of the patients were admitted to inpatient care with a median length of stay of seven days (IQR 3-17). Twenty-four patients (21%) required surgery. Upon discharge, 86.6% of the patients had a mRS grade of 0-2, 3.6 % had a grade of 3, 1.8% had a grade of 4, and 1.8% had a grade of 5. Seven patients (6%) died (mRS grade 6). CONCLUSIONS: Most neurological injuries involve isolated trauma to the head. Fifteen percent of the riders' falls were caused after the horse was impacted by a motor vehicle. Forty percent of the patients require admission and 21% undergo surgery. Ten percent of the patients had a poor mRS grade of 4-6 when discharged.

Impact of the host immune response on the development of equine herpesvirus myeloencephalopathy in horses.

Herpesviruses establish a well-adapted balance with their host's immune system. Despite this co-evolutionary balance, infections can lead to severe disease including neurological disorders in their natural host. In horses, equine herpesvirus 1 (EHV-1) causes respiratory disease, abortions, neonatal foal death and myeloencephalopathy (EHM) in ~10 % of acute infections worldwide. Many aspects of EHM pathogenesis and protection from EHM are still poorly understood. However, it has been shown that the incidence of EHM increases to >70 % in female horses >20 years of age. In this study we used old mares as an experimental equine EHV-1 model of EHM to identify host-specific factors contributing to EHM. Following experimental infection with the neuropathogenic strain EHV-1 Ab4, old mares and yearling horses were studied for 21 days post-infection. Nasal viral shedding and cell-associated viremia were assessed by quantitative PCR. Cytokine/chemokine responses were evaluated in nasal secretions and cerebrospinal fluid (CSF) by Luminex assay and in whole blood by quantitative real-time PCR. EHV-1-specific IgG sub-isotype responses were measured by ELISA. All young horses developed respiratory disease and a bi-phasic fever post-infection, but only 1/9 horses exhibited ataxia. In contrast, respiratory disease was absent in old mares, but all old mares developed EHM that resulted in euthanasia in 6/9 old mares. Old mares also presented significantly decreased nasal viral shedding but higher viremia coinciding with a single fever peak at the onset of viremia. According to clinical disease manifestation, horses were sorted into an EHM group (nine old horses and one young horse) and a non-EHM group (eight young horses) for assessment of host immune responses. Non-EHM horses showed an early upregulation of IFN-α (nasal secretions), IRF7/IRF9, IL-1ß, CXCL10 and TBET (blood) in addition to an IFN-γ upregulation during viremia (blood). In contrast, IFN-α levels in nasal secretions of EHM horses were low and peak levels of IRF7, IRF9, CXCL10 and TGF-ß (blood) coincided with viremia. Moreover, EHM horses showed significantly higher IL-10 levels in nasal secretions, peripheral blood mononuclear cells and CSF and higher serum IgG3/5 antibody titres compared to non-EHM horses. These results suggest that protection from EHM depends on timely induction of type 1 IFN and upregulation cytokines and chemokines that are representative of cellular immunity. In contrast, induction of regulatory or TH-2 type immunity appeared to correlate with an increased risk for EHM. It is likely that future vaccine development for protection from EHM must target shifting this 'at-risk' immunophenotype.

Rapid investigating of phase I metabolites of SR9009 in vitro horse liver microsomes via feature-based molecular networking approach: Potential applications in doping control.

SR9009, a peroxisome proliferator-activated receptor δ (PPARδ) agonist, is known for its potential benefits in energy homeostasis. It failed to receive the United States Food and Drug Administration (USFDA) approval and its illegal distribution has raised concerns. As a result, it has been classified as a prohibited substance by the World Anti-Doping Agency and the International Federation of Horseracing Authorities (IFHA). This study emphasizes the application of the in-silico molecular networking technology to analyze phase I drug metabolites in horses, distinguishing it from conventional methodologies in forensic science. Feature-based molecular networking (FBMN) analysis identified 15 metabolites, with novel major N-dealkylated metabolite (-C8H7NO4S), indicative of diverse metabolic modifications in horse liver microsomes incubation assay. Additionally, a proposed metabolic pathway of SR9009 in the in vitro assay was outlined, including the previously known dehydroxylated metabolite. Finally, the metabolic pathways included in this study were as follows: hydroxylation, dehydrogenation, N-dealkylation dihydroxylation, and combinations. Molecular networking provided insights into MS spectra connectivity, facilitating rapid interpretation and accurate detection of previously undiscovered metabolites. In conclusion, this study contributes to the understanding of SR9009 metabolism in horses and underscores the importance of advanced analytical techniques, such as molecular networking, in enhancing the accuracy and efficiency of metabolite analysis for forensic and doping control purposes.

COVID-19 effects on horses in-transition: A survey analysis of United States equine industry perspectives.

The welfare of unwanted horses presents a significant concern for the equine industry. However, there is a lack of research on how unwanted horses are affected by major crises. The drastic changes that resulted from the COVID-19 pandemic presented ample opportunity to investigate how unwanted horses are impacted by challenging circumstances. Study objectives were to evaluate the COVID-19 pandemic's impact on the unwanted horse population and determine the current perceptions of horses in-transition. A 23-question online survey designed using QualtricsTM was administered electronically to adults living in the United States. Questions pertained to effects on equine ownership, equine management, event participation, and perceptions of unwanted horses. Frequency analysis combined with Chi-squared analyses and analyses of variance identified the impacts of COVID-19 on horse owners, non-horse owners, and equine professionals. From survey results, equine ownership, management practices, and time spent with horses proved to be unaffected (P < 0.001) by the coronavirus pandemic. A decreased ability to participate in equine events was evident across all groups (P ≤ 0.03). Financial hardship, unmanageable behavior, and injury were cited as leading causes for horses becoming "in-transition." Euthanasia was the transitioning method perceived as most accessible, while donation to an equine program was least accessible. Based on results, the COVID-19 pandemic had negligible impact on the number of unwanted horses in the United States. Long-term effects of COVID-19 on equine ownership and management decisions should be considered to provide a deeper base of knowledge for how major crises affect the horse in-transition population.

Occurrence of ultrasonographic assessed placental abnormalities, treatments, pregnancy outcome, and subsequent fertility on a large warmblood stud farm: A retrospective field study.

Little is known about the incidence and outcome of high-risk pregnancies in equine practice and clinical studies on spontaneous occurring placentitis cases and treatments are missing. Therefore, the aims of this retrospective field study were to (1) describe the incidence and severity of ultrasonographic assessed placental abnormalities (UPA) in 4,192 pregnancies on a large commercial warmblood stud farm in 2017 - 2019 and (2) characterize these UPA cases and their pregnancy outcome. UPA severity (Placental abnormality score (PSc) 1-3; low to high), nine treatment regimens (TM1-9) used in UPA cases and treatment duration as well as subsequent fertility were analyzed in the group of UPA mares. The proportion of pregnancies affected by UPA was 4.2 % (n=177/4192). Placental abnormality severity was scored as PSc1 (51.4 %), PSc2 (32.8 %) and PSc3 (15.8 %). The generalized mixed model revealed PSc was affected by mare age and mare status (own pregnancy (OP) or embryo transfer recipient (ER)) (P=0.035) with ER mares having increased PSc compared with mares having their own pregnancy. Abortion occurred in 17/177 (9.6 %) UPA pregnancies. Overall, at the end of the next season, 61.1 % of UPA mares were pregnant, 32.0 % barren, and 6.9 % open (n=175). Pregnancy was established in 62/91(68.1 %) of mares with PSc1, 31/58 (53.4 %) with PSc2 and 14/26 (53.8 %) with PSc3. Most pregnancies were achieved in the first 81/107 (75.7 %) or second 18/107 (16.8 %) inseminated cycle. In conclusion, early detection and treatment of ultrasonographic assessed placental abnormalities can save high-risk pregnancies in > 90 % of cases with a satisfying subsequent fertility.

Short-term equine interaction for reducing test anxiety and facilitating coping skill development in college students during examination periods: A preliminary study.

Mental health concerns are common among college students and more prevalent during examination periods when stress and anxiety are heightened. While the benefits of equine interaction have been reported, the impact of short-term equine interaction on coping skills valuable for managing test anxiety have yet to be studied. Therefore, the purpose of this preliminary study was to evaluate the impact of short-term equine interaction during the college examination period on development of coping skills and anxiety levels associated with testing, both self-reported and those reflected in physiological measures. Full-time college students engaged in a one-hour equine interaction session held on campus at the onset of final exam week. Participants completed a survey targeting perception of anxiety levels and coping skills. Heart rate and salivary cortisol measurements were collected pre- and post-session. Paired t-tests were performed between pre- and post- session measures and correlations between survey answers and physiological measures determined. Significant reductions were found in all areas of anxiety (Overall Academics P = 0.0003, Final Exam Preparation P = 0.0003, Taking Final Exam P = 0.0003). The reduction in exam-related anxiety was positively correlated to cortisol concentrations (r = 0.69, P = 0.03) with lower concentrations (P = 0.05) and increased heart rate (P = 0.003) post-session. Regarding coping skills, improvements were observed post-session in assertiveness (P = 0.008). Skill development coupled with reduced anxiety supports the potential benefits short-term equine interaction can offer students during the high stress periods, but these findings should be confirmed with more in-depth study.

Motor development of stable born healthy foals during the first 24 hours.

The motor development of 14 healthy foals was observed using continuous video-recording from birth to 24 hours of age. An ethogram was made of behaviours of interest with behaviour quantification using CowLog software. Behaviours were divided into six main classes: main activities, attempts to get up or lie down, nursing, playing and other skills, being helped by a human, and the foal not being visible. First-time behaviours (mean, range) of early motor development after birth included going into sternal position (5.4 min, 0-34.5 min), attempting to get up (7.6 min, 0.5-34.6 min), successfully getting up (56.4 min, 27.7 min - 1 h 43.3 min), walking (1 h 1.9 min, 28.1 min - 1 h 43.4 min), nursing (1 h 49.1 min, 1 h 10.3 min - 2 h 29.7 min), shaking (31.9 min, 0.2 min - 2 h 32.7 min), running (2 h 55.6 min, 1h 33.2 min - 6 h 12.1 min), walking backwards (4 h 12.8 min, 56.9 min - 12 h 50.6 min), frolicking (4 h 52.5 min, 2 h 3 min - 15 h 18.8 min), and autogrooming (7 h 30.3 min, 43.3 min - 14 h 40.1 min). All foals made several attempts before they were able to get up for the first time (61.9, 14-103). During the first 24 hours the overall duration of lying down was highest, followed by standing and walking. This information adds to the basic information for assessing normal motor development in these animals, with the potential to identify delayed development.

Equine talocalcaneal subluxation.

A Quarter horse filly was evaluated for a two-week old wound of the right hind cannon bone after the mare stepped on her. A sequestrum was noted radiographically and was surgically debrided. After continued follow-up a left hind varus deformity was noted and was corrected with a lateral transphyseal screw in the distal left tibia. After the varus deformity was improved, a lateromedial radiograph was taken of the distal left tibia and tarsus and talocalcaneal subluxation was seen. The same view was taken of the right tarsus and the same deformity was noted. After finding no case reports on the condition with information on treatment or surgical correction, humane euthanasia was elected. A few months later, the same owner brought a two-month-old colt out of the same mare to the hospital for an elective procedure. Tarsal radiographs were taken due to the owner's previous experience, although the colt had no clinical history or any traumatic incidents. A lateromedial view of the right tarsus revealed talocalcaneal subluxation.

Establishment of a new equine embryo brain primary cell culture with long-term expansion.

Primary cell cultures derived from human embryo lung play a crucial role in virology by aiding virus propagation and vaccine development. These cultures exhibit a notable ability to undergo multiple subcultures, often reaching up to 70 passages. However, finding alternative primary cell cultures with similar longevity and usefulness is challenging. In this study, we introduce a novel primary culture cells derived from equine embryo brain (FEB), which cells exhibited remarkable long-term cultivation potential. The FEB was established and maintained using Sumitomo Nerve-Cell Culture System Comparison studies were conducted with fetal equine kidney cell line (FEK-Tc13) to assess growth rates and subculture longevity. Immunological characterization was performed using neuronal markers to confirm the neural nature of FEB cells. Viral growth assessments were conducted using equine herpesviruses (EHV-1 and EHV-4) to evaluate infectivity and cytopathic effects in FEB cells. PCR analysis and real-time PCR assays were employed to detect viral genomic DNA and transcription activity of EHVs in infected FEB cells. FEB cells demonstrated faster growth rates compared to fetal equine kidney cell line (FEK-Tc13 cells) and exhibited sustained subculture capability exceeding 50 passages. Immunostaining confirmed the glial identity of FEB cells. Both equine herpesviruses 1 and 4 EHV-1 and EHV-4 viruses efficiently replicated in FEB cells, resulting in clear cytopathic effects. PCR analysis detected genomic DNA of EHVs in infected FEB cells, indicating successful viral infection. The establishment of FEB cells with extended subculture capability highlights their potential utility as a model system for studying neural cell biology and viral infections.

Evaluation of gastrointestinal tract lesions and serum malondialdehyde levels after repeated oral administration of phenylbutazone in horses.

Phenylbutazone (PBZ) is a widely used nonsteroidal anti-inflammatory drug for horses. However, because of its gastrointestinal side effects, its administration requires careful attention in veterinary practice. Malondialdehyde (MDA) is a serum biomarker associated with increased damage to the equine gastrointestinal system. This study investigated the hematological effects and alterations in the gastrointestinal tract and assessed serum MDA concentrations following repeated oral PBZ administration at clinical doses. Fourteen horses were randomly divided into control and treatment groups. All horses in the treatment group were administered 4.4 milligrams per kilogram of body weight of PBZ syrup orally twice a day for 7 days, whereas the control group received syrup as a placebo. The development of gastrointestinal side effects was investigated using gastroscopy, abdominal ultrasound, and fecal pH; serum MDA concentrations were assessed using a commercially available enzyme-linked immunosorbent assay kit. Data were compared between PBZ-treated and control horses before and after the treatment period. The treatment group exhibited decreased albumin and total protein concentrations. Moreover, this group exhibited a higher thickness of the right dorsal colon wall (p = 0.03) and had higher scores for squamous gastric ulcers (p = 0.01). Fecal pH was lower in the treatment group than in the control group after PBZ administration (p < 0.01). Although MDA concentrations were higher in the treatment group after PBZ administration, they did not differ significantly from those of the control group. This study highlighted the changes in hematological and gastrointestinal lesions resulting from PBZ administration in horses at clinical doses, even without clinical signs. However, MDA may not be an optimal biomarker for the early detection of gastrointestinal damage due to PBZ treatment in horses.

Arthrospira platensis enriched with Cr(III), Mg(II), and Mn(II) ions improves insulin sensitivity and reduces systemic inflammation in equine metabolic affected horses.

Equine metabolic syndrome (EMS) is a critical endocrine condition in horses, characterized by hyperinsulinemia, hyperlipidemia, and insulin resistance, posing a significant threat to their health. This study investigates the efficacy of supplementing EMS-affected horses with Arthrospira platensis enriched with Cr(III), Mg(II), and Mn(II) ions using biosorption process in improving insulin sensitivity and glucose tolerance, reducing inflammation, and mitigating obesity-related fat accumulation. Our results demonstrate that Arthrospira supplementation reduces baseline insulin and glucose levels, contributing to decreased adipose tissue inflammation. Furthermore, Arthrospira supplementation results in a decrease in body weight and improvements in overall body condition scores and cresty neck scores. Additionally, administration of Arthrospira leads to reduced levels of triglycerides and aspartate aminotransferase, indicating a decrease in hepatic adiposity and inflammation. These findings suggest that Arthrospira, enriched with essential micro- and macroelements, can be an advanced feed additive to enhance insulin sensitivity, promote weight reduction, and alleviate inflammatory processes, thereby improving the overall condition of horses affected by EMS. The use of Arthrospira as a feed additive has the potential to complement conventional management strategies for EMS.

Radiological measurements of the feet of normal Straight Egyptian Arabian horses in Qatar.

The Arabian horse has been identified as carrying a risk locus for equine metabolic syndrome, predisposing this breed to development of laminitis. Radigraphy of the equine foot is widely considered the main diagnostic imaging technique for evaluation of the laminitic horse. Knowledge of 'normal' breed values allows assessment of the degree and severity of radiological changes associated with laminitis. The objective of this study was to investigate the normal values for radiological measurements of the feet of the Straight Egyptian Arabian horse in Qatar. The design was a clinical prospective study. Radiographs of the fore and hind feet of 10 clinically normal adult Straight Egyptian Arabian horses were taken. On the lateromedial views, 17 measurements were taken (13 distances and four angles). On the horizontal dorsopalmar/plantar views, two measurements were taken. On the dorsal 45 degree, proximo-palmarodistal oblique projections, four measurements were taken. Normal reference ranges were reported for radiological measurements of the feet of the Straight Egyptian Arabian horse. Several variables showed significant differences between fore and hind feet, including hoof angle, distal wall thickness, and two proximal inner layer measurements (p < 0.05). In addition, the Straight Egyptian Arabian horse was found to have a number of measurements which varied from previously published reports. The results reported within provide a useful reference for normal radiographic measurements of the Straight Egyptian Arabian horse with relevance for laminitis.

Uncertainties Surrounding Madariaga Virus, a Member of the Eastern Equine Encephalitis Virus Complex.

Background: Madariaga virus (MADV), a member of the eastern equine encephalitis virus (EEEV) complex, circulates in Latin America and exhibits distinct evolutionary and ecological features compared to the North American EEEV. While published data have shed light on MADV ecology, several key aspects remain unknown. Methods: In this study, we compiled data on virus isolation, vector competence, and animal serology collected over six decades in Latin America to identify critical knowledge gaps on MADV transmission and ecology. Results: Specific vertebrate animals serving as amplifying hosts and the mosquito species acting as enzootic and epizootic vectors have not yet been identified. Other aspects that remain unclear are the virus current geographic distribution, the role of equines as hosts in epizootic cycles, and the full impact of MADV on human health in endemic regions. Conclusions: The numerous knowledge gaps surrounding MADV, its widespread distribution in Latin America, and its potential to cause severe disease in animals and humans emphasize the urgent need for increased research efforts, heightened awareness, and intensified surveillance towards this potential emerging threat.