The seroprevalence and risk factors for exposure to Neospora caninum and Neospora hughesi in Ontario broodmares.

The seroprevalence and risk factors for exposure to Neospora caninum and Neospora hughesi in broodmares in Ontario were investigated. Sixty of the 219 (27.4%) study broodmares were seropositive for N. caninum and 65/219 (29.7%) for N. hughesi with cut-offs of ≥1:40 and ≥1:160, respectively. Thirty-one of 63 participating farms (49.2%) had at least 1 broodmare seropositive for N. caninum. Thirty-three of the 63 (52.4%) participating farms had at least 1 broodmare positive for N. hughesi. Risk factors for N. caninum included presence of farm dogs (OR = 6.70; 95% CI = 2.14-20.97; p = 0.001), and high stocking density (OR = 2.83; 95% CI = 1.27-6.30; p = 0.011). Presence of livestock, excluding cattle, was associated with reduced risk of exposure (OR = 0.17; 95% CI = 0.06-0.53; p = 0.002). The only risk factor for exposure to N. hughesi was feeding hay on the ground in the paddock (OR = 4.31; 95% CI = 1.65-11.22; p = 0.003). This study demonstrated widespread exposure to Neospora spp. in broodmares in Ontario.

De novo reconstruction of a functional in vivo-like equine endometrium using collagen-based tissue engineering.

To better understand molecular aspects of equine endometrial function, there is a need for advanced in vitro culture systems that more closely imitate the intricate 3-dimensional (3D) in vivo endometrial structure than current techniques. However, development of a 3D in vitro model of this complex tissue is challenging. This study aimed to develop an in vitro 3D endometrial tissue (3D-ET) with an epithelial cell phenotype optimized by treatment with a Rho-associated protein kinase (ROCK) inhibitor. Equine endometrial epithelial (eECs) and mesenchymal stromal (eMSCs) cells were isolated separately, and eECs cultured in various concentrations of Rock inhibitor (0, 5, 10 µmol) in epithelial medium (EC-medium) containing 10% knock-out serum replacement (KSR). The optimal concentration of Rock inhibitor for enhancing eEC proliferation and viability was 10 µM. However, 10 µM Rock inhibitor in the 10% KSR EC-medium was able to maintain mucin1 (Muc1) gene expression for only a short period. In contrast, fetal bovine serum (FBS) was able to maintain Muc1 gene expression for longer culture durations. An in vitro 3D-ET was successfully constructed using a collagen-based scaffold to support the eECs and eMSCs. The 3D-ET closely mimicked in vivo endometrium by displaying gland-like eEC-derived structures positive for the endometrial gland marker, Fork headbox A2 (FOXA2), and by mimicking the 3D morphology of the stromal compartment. In addition, the 3D-ET expressed the secretory protein MUC1 on its glandular epithelial surface and responded to LPS challenge by upregulating the expression of the interleukin-6 (IL6) and prostaglandin F synthase (PGFS) genes (P < 0.01), along with an increase in their secretory products, IL-6 (P < 0.01) and prostaglandin F2alpha (PGF2α) (P < 0.001) respectively. In the future, this culture system can be used to study both normal physiology and pathological processes of the equine endometrium.

Evaluation of peritoneal l-lactate concentration in horses in the early post-partum period.

BACKGROUND: Peritoneal fluid lactate concentration is an important diagnostic tool in horses with abdominal pain. Information on peritoneal lactate concentrations is lacking following parturition in the mare. OBJECTIVES: To compare blood and peritoneal lactate concentrations in a population of mares within 36 h post-partum, report a normal reference range and identify any impact of retained foetal membranes (RFMs). METHODS: This is a retrospective study evaluating healthy mares from which blood and peritoneal samples had been obtained within 36 h of parturition. Exclusion criteria included signs of abdominal pain within this period. Data was interrogated for normality using a Shapiro-Wilk test. Wilcoxon signed-rank test and Bland-Altman analysis were used to compare blood and peritoneal lactate concentrations. Linear regression was used to compare age and breed data with peritoneal lactate concentrations. Significance was defined as p < 0.05. RESULTS: Forty mares met the inclusion criteria. Mean age was 12.6 ± 4.1 years, and most mares were multiparous (65%). Peritoneal lactate ((1.2 (IQR = 0.9-1.6) mmol/L) was increased compared to blood lactate concentration (0.7 (IQR = 0-1.1)mmol/L; p < 0.001). Plasma total protein (TP) concentrations were 68 (IQR = 64-74) g/L and peritoneal protein concentrations 8 (IQR = 4-9.7) g/L. Six mares developed RFM. The median fold-increase in peritoneal lactate concentration compared to blood lactate concentration was 0.9 (IQR: 0.01-1.7; range: 0-2.5). The reference range for peritoneal fluid lactate concentration was 0-2.5 mmol/L. CONCLUSION: Peritoneal lactate concentrations in healthy post-partum mares remained within the normal reference range and were not influenced by RFM or parturition. Increased peritoneal lactate in this group warrants further investigation.

Micro-CT and histological examination of accessory canals in 34 equine cheek teeth.

Accessory canals and apical deltas have been extensively studied in human dentistry. Their clinical role as a difficult to clean reservoir for bacteria during endodontic treatments has been well described. Many papers describe in detail the pulp anatomy of equine dentition but little attention has been given to their apical ramifications. The goal of this paper is to describe the presence and anatomy of these accessory canals and apical deltas in healthy equine cheek teeth and discuss their possible relevance in the light of equine endodontics. To accomplish this, 15 maxillary and 19 mandibular healthy cheek teeth were collected ranging from Triadan 06 s to 11 s with eruption ages from 4 to 9 years. Their root anatomy was documented in detail based on micro computed tomography images. A subset of 3 teeth also underwent histological examination. Accessory canals were found in all but two teeth examined. Up to 44 accessory canals per tooth have been found with locations ranging from the root furcation down to the apex of the root and with highly variable diameters. Apical deltas in different stages of development were found in 84% of the roots. The presence of accessory canals identified on microCT images could be confirmed using histological examination although some of them were obliterated by reparative dentin. Accessory canals can be found in most equine cheek teeth and add complexity to their endodontic anatomy. This could have important implications for their treatment in case of pulp pathology. In humans, failure to remove bacterial biofilm from such canals has been associated with failure of endodontic treatment. Research on diseased equine teeth is required to gain a better understanding of their clinical relevance in horses.

Horizontal GBR with anorganic equine bone combined with a customized titanium mesh.

This case report describes the fixed rehabilitation of the lower left arch in a patient following an horizontal GBR procedure by means of a customized titanium mesh and a new slow resorption bone substitute of equine origin.

Application of the HRE-S to 140 horses with trigeminal-mediated headshaking and the association of clinical signs with diagnosis, therapy, and outcome.

Background: Horses with trigeminal-mediated headshaking (TMHS) exhibit different headshaking patterns (HSPs), electric shock-like jerking, signs of nasal irritation, and painful facial expressions. The History Rest and Exercise Score (HRE-S) was developed to objectively clarify the severity of the condition in affected horses. This score considers the history and severity of clinical signs at rest and exercise. This study aimed to assess the frequency of different clinical signs and their individual associations with diagnosis, treatment, and outcome in horses diagnosed with TMHS.The clinical records of horses presented with headshaking (HS) at the Clinic for Horses, University of Veterinary Medicine Hannover, between 2006 and 2021 were assessed retrospectively for clinical signs, diagnosis, and treatment. A total of 140 horses were included in the study. Video recordings were evaluated using the HRE-S and compared to the score described by Talbot. Following discharge, owners were interviewed via telephone about the outcome. Correlations between the presence and severity of clinical signs, diagnosis, and outcome were evaluated. Results: The following clinical signs were significantly correlated with a higher HRE-S and grade by Talbot: HS at walk (independently of HSP) (52.9%, 74/140), increased total number of demonstrated HSP (independent of the dominant HSP) (more than one HSP per horse in 91.4%, 128/140), signs of nasal irritation (75.9%, 104/137), painful facial expression (67.8%, 80/118), and electric shock-like jerking (77.5%, 107/138). Diagnosis and outcome do not correlate with the presence of the above-mentioned clinical signs. Conclusion: The HRE-S was confirmed as a valid tool to evaluate disease severity in a cohort of 140 horses with HS. Additionally, clinical signs identified as indicators for higher disease severity may have a stronger negative effect on patient welfare, but they do not correlate with diagnosis or outcome.

Identification of mulberry leaf flavonoids and evaluating their protective effects on H2O2-induced oxidative damage in equine skeletal muscle satellite cells.

Introduction: Horses are susceptible to oxidative stress during strenuous endurance exercise, leading to muscle fatigue and damage. Mulberry leaf flavonoids (MLFs) possess significant antioxidant properties. However, the antioxidant efficacy of MLFs can be influenced by the extraction process, and their impact on H2O2-induced oxidative stress in equine skeletal muscle satellite cells (ESMCs) remains unexplored. Methods: Our study employed three extraction methods to obtain MLFs: ultrasound-assisted extraction (CEP), purification with AB-8 macroporous resin (RP), and n-butanol extraction (NB-EP). We assessed the protective effects of these MLFs on H2O2-induced oxidative stress in ESMCs and analyzed the MLF components using metabolomics. Results: The results revealed that pre-treatment with MLFs dose-dependently protected ESMCs against H2O2-induced oxidative stress. The most effective concentrations were 0.8 mg/mL of CEP, 0.6 mg/mL of RP, and 0.6 mg/mL of NB-EP, significantly enhancing EMSC viability (p < 0.05). These optimized MLF concentrations promoted the GSH-Px, SOD and T-AOC activities (p < 0.05), while reducing MDA production (p < 0.05) in H2O2-induced ESMCs. Furthermore, these MLFs enhanced the gene expression, including Nrf2 and its downstream regulatory genes (TrxR1, GPX1, GPX3, SOD1, and SOD2) (p < 0.05). In terms of mitochondrial function, ESMCs pre-treated with MLFs exhibited higher basal respiration, spare respiratory capacity, maximal respiration, ATP-linked respiration compared to H2O2-induced ESMCs (p < 0.05). Additionally, MLFs enhanced cellular basal glycolysis, glycolytic reserve, and maximal glycolytic capacity (p < 0.05). Metabolomics analysis results revealed significant differences in mulberrin, kaempferol 3-O-glucoside [X-Mal], neohesperidin, dihydrokaempferol, and isobavachalcone among the three extraction processes (p < 0.05). Discussion: Our study revealed that MLFs enhance antioxidant enzyme activity, alleviate oxidative damage in ESMCs through the activation of the Nrf2 pathway, and improve mitochondrial respiration and cell energy metabolism. Additionally, we identified five potential antioxidant flavonoid compounds, suggesting their potential incorporation into the equine diet as a strategy to alleviate exercise-induced oxidative stress.

Plasma and synovial fluid extracellular vesicles display altered microRNA profiles in horses with naturally occurring post-traumatic osteoarthritis: an exploratory study.

OBJECTIVE: The objective of this study was to characterize extracellular vesicles (EVs) in plasma and synovial fluid obtained from horses with and without naturally occurring post-traumatic osteoarthritis (PTOA). ANIMALS: EVs were isolated from plasma and synovial fluid from horses with (n = 6) and without (n = 6) PTOA. METHODS: Plasma and synovial fluid EVs were characterized with respect to quantity, size, and surface markers. Small RNA sequencing was performed, and differentially expressed microRNAs (miRNAs) underwent bioinformatic analysis to identify putative targets and to explore potential associations with specific biological processes. RESULTS: Plasma and synovial fluid samples from horses with PTOA had a significantly higher proportion of exosomes and a lower proportion of microvesicles compared to horses without PTOA. Small RNA sequencing revealed several differentially expressed miRNAs, including miR-144, miR-219-3p, and miR-199a-3l in plasma and miR-199a-3p, miR-214, and miR-9094 in synovial fluid EVs. Bioinformatics analysis of the differentially expressed miRNAs highlighted their potential role in fibrosis, differentiation of chondrocytes, apoptosis, and inflammation pathways in PTOA. CLINICAL RELEVANCE: We have identified dynamic molecular changes in the small noncoding signatures of plasma and synovial fluid EVs in horses with naturally occurring PTOA. These findings could serve to identify promising biomarkers in the pathogenesis of PTOA, to facilitate the development of targeted therapies, and to aid in establishing appropriate translational models of PTOA.

Microvasculature of the suspensory ligament of the equine hind limb.

OBJECTIVE: To describe the microvascular anatomy of the equine hind limb suspensory ligament. ANIMALS: 18 hind limbs harvested from 9 adult horses euthanized for reasons unrelated to lameness. METHODS: A catheter was placed in the transected cranial tibial artery at the level of the mid-distal tibia for each hind limb and used to inject 120 to 150 mL of contrast medium (2 limbs) to identify principal vasculature using contrast-enhanced CT or India ink (11 limbs) to identify microvasculature using the Spalteholz tissue-clearing technique. Routine histologic evaluation was performed on transverse sections from 4 hind limbs. RESULTS: The hind limb suspensory ligament is principally supplied by branches of the medial and lateral plantar metatarsal arteries and, to a lesser extent, the medial and lateral plantar arteries as well as the associated proximal and distal deep plantar arches. A uniformly distributed intraligamentous microvascular supply was observed without relative deficiencies in vascularity between the proximal, midbody, and distal regions. Histologic examination supported these findings, demonstrating a network of connective tissue surrounding and entering the suspensory ligament containing cross-sections of branches of the principal vasculature. CLINICAL RELEVANCE: The equine hind limb suspensory ligament has a uniformly distributed and abundant microvascular supply throughout its length, with no evidence of relative deficiency of vascular supply in any region. A region of hypovascularity does not appear to be a viable explanation for the high rate of injury to and commonality of lameness associated with the proximal hind suspensory ligament in horses.

Welfare assessment of Thoroughbred horses naturally infected with gastrointestinal parasites in Southern Brazil: Quantifying the host-parasite relationship.

Horse welfare assessment (HWA) does not account for individual or herd parasite infection. This study investigated the connection between HWA and individual parasite fecal egg count (FEC) in 90 Thoroughbred horses. All horses were naturally infected with gastrointestinal parasites and were evaluated for individual welfare indicators and FEC monthly, for 12 months. Horses were divided into three groups of 30 mares, 30 foals aged between 13 and 16 months (G2013), and 30 foals aged between two months and one year (G2014). A horse welfare protocol was developed and 1024 assessments were carried out by five trained assessors. FEC ranged from 0 to 5,760 with 98.8 % showing small strongyle eggs. Body condition scores were ideal in 94.4 % of the evaluations (n = 967), and 95.8 % of all horses had good clinical and behavioral indicators. Despite the variation in FEC, the data found no significant association between FEC and the behavioral indicators. The study suggests that FEC alone should not be used as a determinant of welfare when animals are managed with good nutritional and health management practices.

Equine Assisted Occupational Therapy for Children with Adhd (Astride): Protocol Development and Preliminary Study.

Objective: Children with Attention Deficit Hyperactivity Disorder (ADHD) are often referred to Equine-Assisted Services (EAS) for therapy despite lack of validated protocols in the field. This paper reports the development and validation of ASTride (ADHD Skills Therapy): a protocol of Equine-Assisted Occupational Therapy (EAOT) intervention for children aged 6-12 with ADHD. The intervention addresses deficits in cognitive-emotional functions and participation. Method: Phase one of the intervention development includes theoretical framework and core content based on an in-depth review of existing literature. Subsequently, the intervention protocol was revised by a panel of experts. Phase two includes a pilot study, during which five children diagnosed with ADHD (mean age= 10.40 year, SD 2.966) participated in a 12-week EAOT intervention according to the suggested protocol, with pre- and post-assessments conducted. Results: Statistically significant improvements were found in executive functions, as reflected in the Behavioral Regulation Index (BRI) total score. Additionally, hope perception and everyday performance improved following the intervention. Conclusions: Results support the feasibility of ASTride intervention protocol for the improvement of cognitive and emotional functions as well as everyday performance.

Comparison of veterinarians and a deep learning tool in the diagnosis of equine ophthalmic diseases.

BACKGROUND/OBJECTIVES: The aim was to compare ophthalmic diagnoses made by veterinarians to a deep learning (artificial intelligence) software tool which was developed to aid in the diagnosis of equine ophthalmic diseases. As equine ophthalmology is a very specialised field in equine medicine, the tool may be able to help in diagnosing equine ophthalmic emergencies such as uveitis. STUDY DESIGN: In silico tool development and assessment of diagnostic performance. METHODS: A deep learning tool which was developed and trained for classification of equine ophthalmic diseases was tested with 40 photographs displaying various equine ophthalmic diseases. The same data set was shown to different groups of veterinarians (equine, small animal, mixed practice, other) using an opinion poll to compare the results and evaluate the performance of the programme. Convolutional Neural Networks (CNN) were trained on 2346 photographs of equine eyes, which were augmented to 9384 images. Two hundred and sixty-one separate unmodified images were used to evaluate the trained network. The trained deep learning tool was used on 40 photographs of equine eyes (10 healthy, 12 uveitis, 18 other diseases). An opinion poll was used to evaluate the diagnostic performance of 148 veterinarians in comparison to the software tool. RESULTS: The probability for the correct answer was 93% for the AI programme. Equine veterinarians answered correctly in 76%, whereas other veterinarians reached 67% probability for the correct diagnosis. MAIN LIMITATIONS: Diagnosis was solely based on images of equine eyes without the possibility to evaluate the inner eye. CONCLUSIONS: The deep learning tool proved to be at least equivalent to veterinarians in assessing ophthalmic diseases in photographs. We therefore conclude that the software tool may be useful in detecting potential emergency cases. In this context, blindness in horses may be prevented as the horse can receive accurate treatment or can be sent to an equine hospital. Furthermore, the tool gives less experienced veterinarians the opportunity to differentiate between uveitis and other ocular anterior segment disease and to support them in their decision-making regarding treatment.

Effect of odontoplasty on intake, nutrient digestibility, and intake time of tifton hay in ponies.

We aimed to evaluate the effects of dental correction on voluntary nutrient intake, time of intake, apparent digestibility and fecal particle size in Brazilian ponies. Twelve mares, with no history of previous dental treatments, housed in individual pens and fed a diet based on tifton 85 were used. The study comprised two consecutive experimental phases, pre and post dental treatment. Each phase consisted of 15 days for adaptation to the pen and treatment and 5 days for data collection, totaling 20 days. To calculate nutrient intake and digestibility coefficients, samples of diets, leftovers, and feces were analyzed for dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), crude fat (CF), mineral matter (MM), organic matter (OM), and non-fiber carbohydrates (NFC). Food intake time was recorded using a digital video recorder. Fecal particles were analyzed by wet sieving. Dental correction did not influence the intake of DM, CP, NDF, CF, and OM by the animals. However, there was an increase in NFC intake from 0.12 kg to 0.14 kg and in food intake time from 654.50 ± 138.98 to 774.95 ± 167.14. There was a decrease in the digestibility of CP from 0.08 kg to 0.04 kg and an increase in the digestibility of NDF from 0.65 kg to 0.82 kg. Although odontoplasty increased the time of hay intake and the digestibility of the fibrous fractions of the feed of ponies, it did not influence the intake of nutrients and the size of the fecal particles.

IL-1ra gene therapy in equine osteoarthritis improves physiological, anatomical, and biological outcomes of joint degeneration.

OBJECTIVE: To evaluate the effects of a gene transfer approach to IL-1ß inhibition in an equine osteochondral chip fragment model of joint injury using a self-complementary adeno-associated virus with interleukin receptor antagonist transgene cassette (scAAVIL-1ra), as posttraumatic osteoarthritis in horses, similar to people, is a significant clinical problem. ANIMALS: 16 horses were utilized for the study. METHODS: All horses had an osteochondral chip fragment induced arthroscopically in one middle carpal joint while the contralateral joint was sham operated. Eight horses received either scAAVIL-1ra or saline in the osteoarthritis joint. Horses were evaluated over 70 days clinically (lameness, imaging, and biomarker analysis) and euthanized at 70 days and evaluated grossly, with imaging and histopathology. RESULTS: The following findings were statistically significant. Injection of scAAVIL-1ra resulted in high synovial fluid levels of IL-1ra (0.5 to 9 µg/mL) throughout the duration of the experiment (70 days). Over the duration, we observed scAAVIL-1ra to improve lameness (lameness score relative improvement of 1.2 on a scale of 0 to 5), cause suppression of prostaglandin E2 (a relative decline of 30 pg/mL), and result in histological improvement in articular cartilage (decreased chondrocyte loss and chondrone formation) and subchondral bone (less osteochondral splitting and osteochondral lesions). Within the synovial membrane of scAAVIL-1ra-treated joints, we also observed perivascular infiltration with CD3-positive WBCs, suggesting lymphocytic T-cell perivascular infiltration commonly observed with viral transduction. CLINICAL RELEVANCE: These data provide support for further evaluation and optimization of scAAVIL-1ra gene therapy to treat equine osteoarthritis.

Infrared Spectroscopy of Synovial Fluid Shows Accuracy as an Early Biomarker in an Equine Model of Traumatic Osteoarthritis.

Osteoarthritis is a leading cause of lameness and joint disease in horses. A simple, economical, and accurate diagnostic test is required for routine screening for OA. This study aimed to evaluate infrared (IR)-based synovial fluid biomarker profiling to detect early changes associated with a traumatically induced model of equine carpal osteoarthritis (OA). Unilateral carpal OA was induced arthroscopically in 9 of 17 healthy thoroughbred fillies; the remainder served as Sham-operated controls. The median age of both groups was 2 years. Synovial fluid (SF) was obtained before surgical induction of OA (Day 0) and weekly until Day 63. IR absorbance spectra were acquired from dried SF films. Following spectral pre-processing, predictive models using random forests were used to differentiate OA, Sham, and Control samples. The accuracy for distinguishing between OA and any other joint group was 80%. The classification accuracy by sampling day was 87%. For paired classification tasks, the accuracies by joint were 75% for OA vs. OA Control and 70% for OA vs. Sham. The accuracy for separating horses by group (OA vs. Sham) was 68%. In conclusion, SF IR spectroscopy accurately discriminates traumatically induced OA joints from controls.

Conformational Defects in the Limbs of Menorca Purebred Horses and Their Relationship to Functionality.

Limb-conformation defects significantly influence equine performance and welfare, necessitating thorough investigation for effective management. This study examines the prevalence and genetic parameters of 14 limb-conformation defects in Menorca Purebred horses using data from 1120 records (509 animals with an average age of 101.87 ± 1.74 months) collected between 2015 and 2023. Defects were evaluated using a three-class scale by three appraisers, and a Bayesian approach via Gibbs sampling was employed to estimate genetic parameters including gender, birth period, stud selection criteria, evaluation age and appraiser as fixed effects. Splay-footed forelimb and closed hocks were the most prevalent defects (67.20% and 62.53%, respectively). Horses with any of the defects analyzed have been observed to obtain significantly lower scores for both walk and trot. Heritability estimates range from 0.12 (s.d.: 0.025) for closed hock to 0.30 (s.d.: 0.054) for base narrow, confirming the genetic influences on the expression of limb conformation defects. The divergent defect in hind limbs showed the highest genetic correlations with forelimb defects (camped under, -0.69; s.d: 0.32 and camped out, 0.70; s.d: 0.27). The significant genetic correlations between defects highlight the complexity of the relationships, which requires careful consideration.

A Single-Port, Multiple-Access, Custom-Made Device Used in Laparoscopically Assisted Cryptorchidectomy in Standing Horses-A Preliminary Study.

This study evaluates a new multiport device with single access to the abdominal cavity produced with routine hospital supplies that could be applied to laparoscopically assisted cryptorchidectomy in standing horses. Initially, the new device was evaluated on five cadavers of bovine fetuses (n = 5), placed assisted in a minilaparotomy performed in the flank region. Subsequently, the device was evaluated in four cryptorchid horses treated during the hospital routine. During the evaluation of the new device, the possibilities of exploring the abdominal cavity, inspection, and intra-abdominal manipulation with two Babcock forceps were verified. The possibilities were described, and surgical time data were recorded and analyzed using descriptive statistics. In the cadavers, a wide exploration of the abdominal cavity was possible, with a laparoscopic inspection through the right paralumbar fossa and manipulation of intra-abdominal structures with Babcock forceps inserted by the new device. In cryptorchid horses, laparoscopically assisted cryptorchidectomy with a new device was feasible in two patients, and in the others, it allowed the diagnosis of adhesions and ectopic locations in the inguinal region of testicles retained in the cavity. Therefore, the new device was efficient in exploring the inguinal region of cryptorchid horses in the standing position. The present study is preliminary and can support future studies that aim to improve the developed prototype.

Classification performance of sEMG and kinematic parameters for distinguishing between non-lame and induced lameness conditions in horses.

Despite its proven research applications, it remains unknown whether surface electromyography (sEMG) can be used clinically to discriminate non-lame from lame conditions in horses. This study compared the classification performance of sEMG absolute value (sEMGabs) and asymmetry (sEMGasym) parameters, alongside validated kinematic upper-body asymmetry parameters, for distinguishing non-lame from induced fore- (iFL) and hindlimb (iHL) lameness. Bilateral sEMG and 3D-kinematic data were collected from clinically non-lame horses (n = 8) during in-hand trot. iFL and iHL (2-3/5 AAEP) were induced on separate days using a modified horseshoe, with baseline data initially collected each day. sEMG signals were DC-offset removed, high-pass filtered (40 Hz), and full-wave rectified. Normalized, average rectified value (ARV) was calculated for each muscle and stride (sEMGabs), with the difference between right and left-side ARV representing sEMGasym. Asymmetry parameters (MinDiff, MaxDiff, Hip Hike) were calculated from poll, withers, and pelvis vertical displacement. Receiver-operating-characteristic (ROC) and area under the curve (AUC) analysis determined the accuracy of each parameter for distinguishing baseline from iFL or iHL. Both sEMG parameters performed better for detecting iHL (0.97 ≥ AUC ≥ 0.48) compared to iFL (0.77 ≥ AUC ≥ 0.49). sEMGabs performed better (0.97 ≥ AUC ≥ 0.49) than sEMGasym (0.76 ≥ AUC ≥ 0.48) for detecting both iFL and iHL. Like previous studies, MinDiff Poll and Pelvis asymmetry parameters (MinDiff, MaxDiff, Hip Hike) demonstrated excellent discrimination for iFL and iHL, respectively (AUC > 0.95). Findings support future development of multivariate lameness-detection approaches that combine kinematics and sEMG. This may provide a more comprehensive approach to diagnosis, treatment, and monitoring of equine lameness, by measuring the underlying functional cause(s) at a neuromuscular level.

Expression of recombinant Florida clade 2 hemagglutinin in baculovirus expression system: A step for subunit vaccine development against H3N8 equine influenza virus.

Background: Equine influenza (EI) is a transmissible viral respiratory sickness of the Equidae family. Two viruses, H7N7 and H3N8 caused EI; however, H7N7 has not been detected for decades. H3N8 has circulated and bifurcated into Eurasian and American lineages. The latter subsequently diversified into Kentucky, South America, and Florida sub-lineages. Florida clade 1 (FC1) and Florida clade 2 (FC2) strains are the only circulating EI viruses (EIVs) in the meantime. Immunization is considered the major means for the prevention and control of EI infection. Using disparate technologies and platforms, several vaccines have been developed and commercialized. According to the recommendations of the World Organization for Animal Health (WOAH), all commercial vaccines shall comprise representatives of both FC1 and FC2 strains. Unfortunately, most of the commercially available vaccines were not updated to incorporate a representative of FC2 strains. Aim: The purpose of this research was to develop a new EI vaccine candidate that incorporates the hemagglutinin (HA) antigen from the currently circulating FC2. Methods: In this study, we report the expression of the full-length recombinant HA gene of FC2 in the baculovirus expression system. Results: The HA recombinant protein has been proven to maintain its biological characteristics by hemadsorption (HAD) and hemagglutination tests. Moreover, using a reference-specific serum, the specificity of the HA has been confirmed through the implementation of immunoperoxidase and western immunoblotting assays. Conclusion: In conclusion, we report the expression of specific biologically active recombinant HA of FC2, which would act as a foundation for the generation of an updated EI subunit or virus vector vaccine candidates.