Overexpression of the key metabolic protein Carnitine Palmitoyl Transferase 1A (CPT1A) in equine sarcoid.

The equine sarcoid is the most common skin neoplasia of fibroblastic origin in horses, characterized by an excessive accumulation of extracellular matrix produced by sarcoid fibroblasts under hypoxic condition. Neoplastic cells can adapt to hypoxia by using alternative energy sources, particularly those that arise from fatty acid oxidation (FAO). The Carnitine Palmitoyl Transferase 1A (CPT1A) belongs to Carnitine System (CS) and promotes the entrance of fatty acids into the mitochondria for ß-oxidation. In this study, CPT1A expression was comparatively addressed in 25 equine sarcoids and 5 normal skin samples using immunohistochemistry (IHC). Specificity of CPT1A antibody was validated by Western Blotting (WB). In normal skin samples IHC staining was weak and mainly confined to basal epidermis and few dermal fibroblasts. Sarcoid fibroblast exhibited a strong cytoplasmic and nuclear signal in 60% of the tumor samples. Cytoplasmic CPT1A expression in sarcoid fibroblasts indicates that the protein is actively involved in metabolic reprogramming processes. Nuclear CPT1A expression suggests that the protein may also be involved in the regulation of neoplastic proliferation.

Exploring Histoplasma species seroprevalence and risk factors for seropositivity in The Gambia's working equid population: Baseline analysis of the Tackling Histoplasmosis project dataset.

Introduction: Exposure rates to Histoplasma species, the causative agent of equine epizootic lymphangitis (EL), are unknown amongst working equids in The Gambia. The primary aims of this study were to estimate anti-Histoplasma antibody seroprevalence in the equid population in rural The Gambia and to explore risk factors for seropositivity. Methods: A nationwide cross-sectional study was conducted (February-July 2022), representing baseline measurements of a longitudinal cohort study. Horses (n = 463) and donkeys (n = 92) without EL signs were recruited in 18 study sites. Following informed owner consent, equid clinical and management data were recorded. Blood samples were collected by jugular venepuncture, and sera were subject to the IMMY Latex Agglutination Histoplasma test (LAT). Seropositivity risk factors were explored by multi-level, multivariable logistic regression analysis. Study site and household variance were described using a latent-variable approach. Whole blood DNA extractions were subject to nested ITS-PCR to detect Histoplasma capsulatum var. farciminosum (HCF), and agreement with LAT results was measured using Cohen's kappa statistic. Results: Anti-Histoplasma antibody seroprevalence in horses and donkeys was 79.9% [95% confidence interval (CI) 76.0-83.5%] and 46.7% (95% CI 36.3-57.4%), respectively. In horses, two multivariable models explained the maximum amount of data variability. Model 1 demonstrated increased odds of seropositivity in mares [odds ratio (OR) = 2.90 95% CI 1.70-4.95, p < 0.001] and decreased odds in horses <2.5 years (OR = 0.46 95% CI 0.22-0.95, p = 0.04; reference: ≥4.5 years). Model 2 demonstrated increased odds in horses recruited during the rainy season (OR = 2.03 95% CI 1.08-3.84, p = 0.03) and those owned by farmers reporting previous EL in their equids (OR = 1.87 95% CI 1.04-3.37, p = 0.04). Decreased odds were measured in horses <2.5 years (OR = 0.37 95% CI 0.18-0.78, p = 0.01) and horses reported to transport firewood (OR = 0.45 95% CI 0.28-0.74, p = 0.001). On multivariable analysis of donkeys, decreased odds of seropositivity were demonstrated amongst donkeys owned by households which also owned horses (OR = 0.23 95% CI 0.06-0.85, p = 0.03). HCF infection prevalence in horses and donkeys was 22.0% (n = 102/463, 95% CI 18.3-26.1%) and 5.4% (n = 5/92, 95% CI 1.8-12.2%), respectively. No significant agreement was measured between LAT and nested ITS-PCR results (κ < 0.00). Conclusion: High Histoplasma spp. exposure was demonstrated amongst equids in The Gambia. Investigation of risk factors, including equid husbandry and management strategies, as well as geoclimatic variations, is warranted. Outcomes may inform sustainable and equitable EL control strategies in The Gambia and comparable settings worldwide.

Rabies in equids in Sudan.

Rabies is endemic in Sudan with continuing outbreaks occurring annually, the most common animals affected are dogs, followed by goats and equids. This work focused on equid rabies, to elucidate the current situation of the disease through analysis of reports of equid rabies outbreaks in Sudan during 2010-2022 supported by laboratory confirmation of the disease. During the study period, 66 animals were affected during 35 equid rabies outbreaks. The highest incidences were found in Al Gezira (30.3%), followed by Darfur (24.2%) and Kordofan (15.2%). The highest incidence rate was observed during 2018 (33.3%), followed by 2015 (16.7%). Within seasons, the highest incidence rate was reported during October - December (33.3%), followed by July - September (30.3%). Chi-square analysis revealed a significant correlation between rabid animals and year, season, and state. Wald statistics demonstrated that year and season had a significant association with the disease. Virus antigen was identified (72.2%) in brain tissues using the fluorescent antibody test. Viral nucleic acid was amplified (n = 6) with a reverse transcriptase polymerase chain reaction assay.Contribution: As equids are kept in close contact with humans and other animals in the country, according to the present investigation, equid rabies in Sudan is a potential public health concern, emphasising the importance of implementing effective control measures.

Technique description and outcome evaluation of Thoroughbred racehorses following soft palate thermocautery performed under standing sedation.

BACKGROUND: Palatal dysfunction (PD), which encompasses palatal instability (PI) and intermittent dorsal displacement of the soft palate, is the most common performance-limiting upper respiratory tract obstruction in young Thoroughbred racehorses. Soft palate thermocautery (SPT) performed under general anaesthesia is a routinely performed procedure for PD in some countries, but the procedure for and outcome of SPT performed under standing sedation has not been published. OBJECTIVES: (1) To describe a technique for SPT performed under standing sedation; and (2) to assess post-operative performance in horses compared to controls using the Racing Post rating (RPR), British Horseracing Authority official rating (OR), Performance Index and Earnings. STUDY DESIGN: Retrospective case series. METHODS: Medical records were reviewed for all horses that had SPT performed under standing sedation following topical and local infusion of lidocaine hydrochloride into the rostral soft palate, and that were identified to have PI by overground endoscopy. Two matched controls were identified for each case. The median RPR, OR, Performance Index and Earnings for the three pre-operative and three post-operative races were compared. RESULTS: No significant differences were identified between the SPT (n = 23) and Control groups (n = 46) for baseline characteristics or outcomes. MAIN LIMITATIONS: Retrospective study design, small sample size. CONCLUSIONS: No significant differences in racing performance were identified between horses that had SPT performed under standing sedation and controls.

The prognostic importance of physiological and biochemical parameters in horses afflicted with colic.

Background: Colic, a primary cause of illness and death in horses, necessitates the development of improved prognostic tools. Aim: The aim of this study was to investigate the prognostic significance of physiological and biochemical parameters in horses suffering from colic. Methods: A comprehensive clinical evaluation of 117 horses included assessment of heart rate, mucous membranes, capillary refill time, rectal temperature, respiratory rate, gut motility, reflux, and limb pulse strength. Results: Stomach reflux, absence of gut noises, and increased heart rate (mean increase of 12 bpm) strongly correlate with a poor prognosis. Prolonged capillary refill time (mean increase of 3 seconds), rectal temperature (over 38.5°C), elevated packed cell volume (mean increase of 4%), and blood lactate levels (mean increase of 5 mmol/l) underscore the significance of these markers. Notably, blood lactate (p < 0.001), gut noises (p < 0.05), and heart rate (p < 0.001) demonstrate the highest predictive significance based on statistical analysis. Conclusion: Future research should investigate the prognostic potential of additional parameters and assess the impact of recommended treatments on colic prognosis. This data-driven study emphasizes the critical role of early recognition and thorough assessment in colic cases, offering vital insights into improving equine healthcare and mitigating mortality rates.

Initial development of a rapid, portable, stall-side ELISA for the measurement of equine adrenocorticotropic hormone.

Pituitary pars intermedia dysfunction (PPID) is a neurodegenerative disease of senior horses. Loss of dopaminergic inhibition of the melanotropes of the pars intermedia leads to increased concentrations of pro-opiomelanocortin (POMC)-derived peptides. Diagnosis is challenging due to pre-analytical variables, such as sample storage, handling, and time to analysis. Our objective was to develop an ELISA for ACTH measurement, which could ultimately form the basis for a stall-side equine ACTH test. We selected 2 ACTH-specific monoclonal antibodies, CBL57 and EPR20361-248, based on the recognition of separate epitopes, strong and rapid color change, and minimal background interference, including no cross-reactivity with themselves, each other, and the test reagents. CBL57 was chosen as the detection antibody (or secondary antibody). EPR20361-248, functionalized on superparamagnetic iron oxide beads, was chosen as the capture antibody (or primary antibody) to bind ACTH in plasma. The incorporation of magnetic beads marks the initial stage in establishing a platform that could potentially be utilized in the field, similar to other stall-side tests. The concentrations of antibodies, magnetic beads, and incubation durations were optimized. Our immunoassay detected unglycosylated rat recombinant ACTH. Further studies are ongoing to optimize and validate our assay using equine plasma and serum samples.

Equine Doping Controls of Thymosin ß $$ \beta $$ 4: A Population Study and Strategy for Misuse Detection.

Thymosin ß $$ \beta $$ 4 (TB4) is a ubiquitous, highly conserved and abundant peptide among mammals with a critical role in cytoskeleton organization. In spite of its yet non-authorized use as a medicine and being forbidden by the IFHA, the FEI, and the WADA, intelligence and doping control laboratories reported numerous products available online claiming to contain a synthetic acetylated fragment of TB4 or TB4 itself, promoted as a growth factor with regenerative properties. In this article, the first estimation of the endogenous TB4 concentration in racing horses' blood samples was performed through a population study. We reveal that this concentration does not significantly depend on gender, age, nor horse breed. We highlight that the TB4 concentration increases significantly and rapidly in plasma stored at 4°C when not separated from blood cells due to cell lysis. Finally, we also demonstrate that the detection of a non-natural synthesis impurity is possible in equine plasma after a single dose administration of a TB4 containing product to a horse.

Treatment of experimental hyperchloremic metabolic acidosis in horses with enteral electrolyte solution containing sodium acetate.

Introduction: In adult horses, the development and evaluation of enteral electrolyte solutions containing sodium acetate for correcting hyperchloremic metabolic acidosis are still lacking, although these electrolyte and acid-base imbalances are commonly observed. The objective of this study was to evaluate the alkalinizing effect of two enteral electrolyte solutions containing different concentrations of acetate, administered via nasogastric tube in continuous flow, in adult horses with experimental hyperchloremic metabolic acidosis. Methods: Six mares aged between 3 and 10 years were used in a 6×2 crossover design, with each animal receiving both treatments. The horses were subjected to a protocol to induce hyperchloremic metabolic acidosis. They then received one of two treatments: HighAcetate (81.4 mmol/L) and LowAcetate (22.7 mmol/L) at an infusion rate of 15 mL/kg/h for 12 h. Plasma, serum, and urinary biochemical assessments; hematocrit; urinary volume, pH, and specific gravity; and blood gas analysis were measured at the following time points: T-12 h (beginning of the 12-h fast), T0h (end of fasting and beginning of the acidosis induction phase), every 2 h during the hyperchloremic metabolic acidosis induction phase (Ti2h and Ti4h), every 2 h during the 12-h enteral hydration phase (Tt2h, Tt4h, Tt6h, Tt8h, Tt10h, and Tt12h), with one sample taken at T24h (24 h after the start of acidosis induction) and another at T36h (36 h after the start of acidosis induction). Data were analyzed using descriptive statistics and analysis of variance based on a factorial design of repeated measures, with Tukey's post-hoc test or the Kruskal-Wallis test with Dunn's post-hoc test for non-parametric tests. Results: At the end of the induction phase, the animals developed moderate to severe hyperchloremic metabolic acidosis. The HighAcetate solution effectively corrected electrolyte and acid-base imbalances before the end of the treatment phase (Tt12h), while the LowAcetate solution was not effective in correcting those changes. Conclusion: The HighAcetate (81.4 mmol/L) solution is deemed an effective and safe alternative for the treatment of hyperchloremic metabolic acidosis in horses.

Treatment of equine keratomycosis, part 2: reporting common systemic and ocular complications to enhance decision-making for equine clinicians and owners.

OBJECTIVE: The primary aims were to provide descriptive data on systemic and ocular complications associated with the treatment of equine keratomycosis. The secondary aims were to determine if complication rates differ between treatment types or are related to surgical and anesthetic factors. METHODS: The study was a retrospective evaluation of 126 cases between 2004 and 2020 with confirmed fungal infection and recorded incidence of complications during hospitalization and surgical intervention if pursued. Additional information recorded included the size of the donor graft if utilized, time under general anesthesia, and prescribed medications. Fisher exact and χ2 tests were used to evaluate complication frequency differences. Logistic regression models determined the effects of donor graft size, anesthesia time, and duration of hospitalization on complication rates. RESULTS: Hypercreatininemia followed by colic were the most frequent systemic complications in the medical treatment group. Colic and conjunctival graft dehiscence were the most frequent systemic and ocular complications in all surgery treatment groups, respectively. The frequency of systemic complications did not vary between treatment groups, although colic rates were significantly higher following general anesthesia. All colic cases were resolved with supportive care. Donor graft size, anesthesia time, and duration of hospitalization did not impact complication frequency. The mainstays of medical therapy included topical fluoroquinolones, topical voriconazole, topical atropine, oral NSAIDS, and oral gastroprotectants. CONCLUSION: Treatment of keratomycosis is associated with a range of complications. Colic remains a significant risk, with increased frequency reported following general anesthesia. CLINICAL RELEVANCE: Reporting common complications associated with treatment aids in decision-making for equine clinicians and owners.

New Transcriptomic Biomarkers for Detection of the Recombinant Human Erythropoietin (rHuEPO) MirCERA in Horses.

Detection and monitoring of biomarkers related to doping is particularly suitable for the development of analytical strategies dedicated to indirect detection of banned substances. Previous studies in horses have already allowed the investigation of transcriptomic biomarkers in equine blood associated with reGH and rHuEPO administrations. Our most recent developments continue to focus on the discovery and monitoring of transcriptomic biomarkers for the control of ESAs, and a collaborative study with WADA-accredited doping control laboratories has recently been initiated to conduct a pilot study. In humans, three mRNAs (ALAS2, CA1, and SLC4A1) were previously observed to be differentially expressed after blood doping and were associated with immature red blood cells, the so-called circulating reticulocytes. In horses, circulating reticulocytes are rarely observed even after rHuEPO administration. With the improved primers that detect the equine orthologues of the human mRNAs from the ALAS2, CA1, and SLC4A1 genes, we can now report the first evidence of the detection of the three biomarkers in equine blood. In addition, an upregulation of the mRNA levels of the three genes was observed after analysis of blood samples collected from MirCERA-treated animals, with kinetics similar to those previously documented in humans. Our data suggest that ALAS2 and CA1 are promising indirect biomarkers for the detection of recombinant EPO abuse in horses, as observed in humans.

Maturational competence of equine oocytes is associated with alterations in their 'cumulome'.

Assisted reproductive technologies are an emerging field in equine reproduction, with species-dependent peculiarities, such as the low success rate of conventional IVF. Here, the 'cumulome' was related to the developmental capacity of its corresponding oocyte. Cumulus-oocyte complexes collected from slaughterhouse ovaries were individually matured, fertilized by ICSI, and cultured. After maturation, the cumulus was collected for proteomics analysis using label-free mass spectrometry (MS)-based protein profiling by nano-HPLC MS/MS and metabolomics analysis by UPLC-nanoESI MS. Overall, a total of 1671 proteins and 612 metabolites were included in the quantifiable 'cumulome'. According to the development of the corresponding oocytes, three groups were compared with each other: not matured (NM; n = 18), cleaved (CV; n = 15), and blastocyst (BL; n = 19). CV and BL were also analyzed together as the matured group (M; n = 34). The dataset revealed a closer connection within the two M groups and a more distinct separation from the NM group. Overrepresentation analysis detected enrichments related to energy metabolism as well as vesicular transport in the M group. Functional enrichment analysis found only the KEGG pathway 'oxidative phosphorylation' as significantly enriched in the NM group. A compound attributed to ATP was observed with significantly higher concentrations in the BL group compared with the NM group. Finally, in the NM group, proteins related to degradation of glycosaminoglycans were lower and components of cumulus extracellular matrix were higher compared to the other groups. In summary, the study revealed novel pathways associated with the maturational and developmental competence of oocytes.

A platform for the rapid screening of equine immunoglobins F (ab)2 derived from single equine memory B cells able to cross-neutralize to influenza virus.

Single B cells-based antibody platforms offer an effective approach for the discovery of useful antibodies for therapeutic or research purposes. Here we present a method for screening equine immunoglobins F(ab)2, which offers the potential advantage of reacting with multiple epitopes on the virus. Using equine influenza virus (EIV) as model, a hemagglutinin (HA) trimer was constructed to bait B cells in vaccinated horses. We screened 370 HA-specific B cells from 1 × 106 PBMCs and identified a diverse set of equine variable region gene sequences of heavy and light chains and then recombined with humanized Ig Fc. Recombinant equine Ig was then self-assembled in co-transfected 293 T cells, and subsequently optimized to obtain HA binding B-cell receptor (s). The recombinant antibodies exhibited a high binding affinity to the HA protein. Antibody H81 exhibited the highest cross neutralizing activities against EIV strains in vitro. Furthermore, it effectively protected EIV-challenged mice, resulting in significantly improved survival, reduced pulmonary inflammation and decreased viral titers. In silico predication identified a functional region of H81 comprising 27 key amino acids cross the main circulating EIV strains. The 12 amino acid residues in this region with the highest binding affinities were screened. Notably, the predicted epitopes of H81 encompassed the documented equine HA receptor binding site, validating its cross-neutralization. In summary, a rapid platform was successfully established to investigate the profiling of equine antigen-recognizing receptors (BCRs) following infection. This platform has the potential to optimize the screening of virus-neutralizing antibodies and aid in vaccine design.

Comparison of distal extremity magnetic resonance imaging findings in Australian racing Standardbreds and Thoroughbreds.

INTRODUCTION: Foot-related lameness is common in Standardbred and Thoroughbred racehorses. Magnetic resonance imaging (MRI) provides information on both the soft tissue and osseous structures of the distal extremity. The aim of this study was to determine whether abnormalities identified on distal limb MRI differed between these two racehorse populations. MATERIALS AND METHODS: The records of Standardbred and Thoroughbred racehorse distal extremity MRI examinations conducted at Camden Equine Centre and Ballarat Veterinary Practice between 2013 and 2023 were retrieved. Horses were included if pain causing lameness had been localised to the foot using perineural or intra-articular analgesia. MRI abnormalities of the structures within the distal extremity were recorded and comparisons made between the two groups. RESULTS: There was a significant difference in the rate of occurrence of certain MRI abnormalities between the Thoroughbred and Standardbred racehorse populations, including distal phalangeal fractures, proximal phalangeal short tau inversion recovery (STIR) hyperintensity (distal aspect), the presence and severity of ungular cartilage ossification, abnormalities of the navicular bone distal border synovial invaginations/vascular channels and navicular bone distal border fragments. The presence of distal extremity STIR hyperintensity and fractures was significantly higher in Standardbred compared to Thoroughbred racehorses when these findings were grouped and considered in combination. CONCLUSION: There were many similarities in the distal extremity MRI abnormalities identified in lame Australian Standardbred and Thoroughbred racehorses. However, distal phalangeal fractures, ungular cartilage ossification and distal extremity STIR hyperintensity were identified in significantly more in Standardbred than Thoroughbred racehorses.

Quadricuspid aortic valve in a 16-year-old Quarter horse.

A 16-year-old horse, 400 kg, male, Quarter horse gelding was examined for a cardiac murmur that had not been previously heard. Physical examination revealed a regularly irregular pulse and a grade III/VI, decrescendo, diastolic murmur with a point of maximum intensity over the left heart base. Base-apex standard electrocardiographic examination at rest showed sinus rhythm with second-degree atrio-ventricular blocks. Echocardiography identified the presence of four aortic valve cusps, two equal larger and two unequal smaller cusps. Color flow Doppler examination showed a diastolic regurgitant jet emerging from the central region of closed aortic valve. Based on clinical and echocardiographic findings a diagnosis of mild to moderate aortic valve insufficiency secondary to quadricuspid aortic valve was made. This report describes a rare congenital heart defect that can be detected by transthoracic echocardiography in the horse.

Fecal microbiome and functional prediction profiles of horses with and without crib-biting behavior: A comparative study.

Crib-biting is a stereotyped oral behavior with poorly understood etiology and pathophysiology. The relationship between the gut microbiome and brain function has been described in behavioral disorders such as schizophrenia, depression and anxiety in humans. In horses, studies of behavioral problems and the microbiome are very limited. This study aimed to characterize the fecal microbiome and the predicted functional profile of horses with and without aerophagia. Fecal samples were collected from 12 Colombian Creole Horses of both sexes, divided into two groups: group 1, composed of six horses with crib-biting (3 females and 3 males), average body weight of 330 ± 10 kg, age of 7.0 ± 1.2 years and body condition score (BCS) of 5/9 ± 1 and group 2, consisting of six horses without crib-biting (3 females and 3 males), average body weight of 335 ± 5 kg, age 6.5 ± 1 years and BCS of 6/9 ± 1. From each horse in both groups fecal total DNA was obtained and 16S ribosomal RNA gene amplicons were sequenced to characterize the bacterial community structure. Community structure and differential abundance analyses revealed significant differences between the two conditions (p < 0.05). Specifically, the fecal microbiota at the family level in crib-biting horses, showing a decrease in Bacteroidales and an increase in Bacillota and Clostridia, differed from that of healthy horses without crib-biting, consistent with findings from previous studies. Furthermore, metagenome prediction suggests metabolic profile changes in bacterial communities between both conditions in horses. Further studies are required to validate the role of the microbiota-gut-brain axis in the etiology of crib-biting and other abnormal and stereotyped behaviors.

Significance of Anaplasma phagocytophilum, Babesia caballi, and Theileria equi as etiologic agents in horses with clinical manifestations from the metropolitan area of Rio De Janeiro, Brazil.

Equine Piroplasmosis (EP) and Equine Granulocytic Anaplasmosis (EGA) are diseases that affect horses, transmitted by ixodid ticks, causing a nonspecific febrile syndrome. Equine Piroplasmosis is endemic in Brazil, and most horses are in enzootic stability. Serological and molecular studies carried out on horses in Brazil have shown the presence of Anaplasma phagocytophilum, however, the clinical relevance of this infection has not yet been established. The present study aims to evaluate the importance of Babesia caballi, Theileria equi, and A. phagocytophilum as etiological agents in horses with clinical manifestations suggestive of these diseases in the metropolitan mesoregion of Rio de Janeiro. A total of 45 animals with clinical signs were submitted to DNA extraction followed by qPCR test. Anaplasma phagocytophilum, Neorickettsia risticii and Theileria haneyi were not found in any of the horses with clinical signs, however 62.2% were infected with at least one agent of EP. Theileria equi was the most frequent etiologic agent (35.5%), followed by coinfection (15.5%) and B. caballi (11.2%). These results suggest that A. phagocytophilum has minor clinical importance in the region, while EP is frequently found in symptomatic horses, representing an important differential diagnosis in suspected cases.

Effect of intrafollicular administration of PGE2 or PGF2α in early estrus on ovulation, hemorrhagic anovulatory follicles formation, progesterone secretion and pregnancy outcome in the mare.

This experiment was performed to evaluate whether intrafollicular treatment of PGE2 or PGF2α administered in early estrus would induce normal ovulation, progesterone production (Experiment 1) and pregnancy (Experiment 2). In Experiment 1, mares in estrus after 2 days of endometrial edema were injected in all largest dominant follicles (28-35 mm in diameter) with 0.5 mL of sterile water containing 500 µg PGE2 (n = 6), 125 µg PGF2α (n = 6) or placebo (n = 7) (Hour 0). Ultrasound examinations were performed daily, until ovulation or anovulation was detected, and daily blood samples were taken for 8 days. In Experiment 2, mares with a dominant follicle ≥35 mm after at least three days of slight-to-moderate endometrial edema, were injected with 500 µg PGE2 diluted in 0.5 mL of sterile water for injection in the follicle (PGE2 group; n = 9 mares and 11 dominant follicles). No puncture was performed in the control group (n = 9 mares and 11 dominant follicles). Mares from both groups were inseminated. In Experiment 1, all mares (6/6) in the PGE2 group ovulated within 24 h of treatment. The mean interval from intrafollicular injection to ovulation was shorter (P < 0.001) in PGE2 mares (24 ± 0 h) than in control mares (77 ± 9 h). Mares from the PGF2α group developed hemorrhagic anovulatory follicles (HAF) more often (7/7) than control mares (2/7); P < 0.05). The progesterone concentration in mares from the PGF2α group was lower (P < 0.004) than control mares in the early post-ovulatory period. The first significant increase in post-ovulatory progesterone concentration occurred earlier (P < 0.05) in mares from the control group than in mares from the PGF2α and PGE2 groups. In Experiment 2, more mares from the control group (7/9, 78 %) became pregnant than from the PGE2 group (2/9, 22 %) (P = 0.015). In conclusion, PGE2 alone induced follicle collapse in all treated mares within 24 h of administrations, while PGF2α blocked ovulation and induced formation of HAFs. However, the post-ovulatory rise in progesterone production was delayed and the fertility reduced in mares with ovulation induced by PGE2 compared to control mares.

Unveiling directional physiological coupling in human-horse interactions.

This research investigates the human-horse bond, aiming to unveil the physiological mechanisms regulating interspecies interactions. We hypothesized observing a physiological synchronization in human-horse dynamics, akin to human interactions. Through time-frequency Granger causality analysis of heart rate variability (HRV) and behavioral data, this study reveals the establishment of bidirectional synchronization in HRV between humans and horses. The coupling directionality is influenced by behavior and familiarity. In exploration scenarios led by horses, bidirectional interactions occur, particularly with familiar individuals. Conversely, during human-led activities such as grooming, physiological connectivity direction varies based on the familiarity level. In addition, the methodology allows in-depth analysis of sympathetic and parasympathetic nervous system contributions, highlighting their intricate role in the human-horse relationship. Such a physiological coupling estimate, correlated with behavioral data, provides a quantitative tool applicable across contexts and species This holds significant promise for assessing animal-assisted therapies and for applications in sports and various animal-related domains.

High-Throughput Equine Doping Controls on a Trapped Ion Mobility Quadrupole-Time-of-Flight Mass Spectrometer: Technical Considerations of dia/slice/prmPASEF Applied to the Long-Term Detection of Monoclonal Antibodies.

Data-independent acquisition (DIA) methods employing a scanning quadrupole were recently described across multiple platforms. These strategies display remarkable performances in untargeted proteomics studies thanks to rapid duty cycles, leading to ultrashort liquid chromatography gradients while maintaining enough data points per peaks when coupled to fast-scanning mass analyzer. In this article, we perform the evaluation of three data acquisition strategies named diaPASEF,slicePASEF, and prmPASEF on a trapped ion mobility spectrometry quadrupole-time-of-flight (TIMS-Q-TOF) mass spectrometer for high-throughput doping control screening analyses. We report that slicePASEF outperforms diaPASEF and is almost as sensitive as prmPASEF in detecting humanized monoclonal antibodies for several weeks in equine plasma after administration. We observed that diaPASEF is still providing the best performances in untargeted proteomics studies employing high amounts of input materials, which is linked with the high complexity of slicePASEF data and current processing algorithms.

Discovery of Biomarkers of a Recombinant Human Erythropoietin Administration to Thoroughbred Geldings by Label-Free Proteomics.

Erythropoiesis-stimulating agents (ESAs) continue to be a significant threat to the integrity of human and equine sports. Besides conventional direct testing, monitoring the biomarkers associated with the effects of ESAs may provide a complementary approach via indirect detection to enhance doping control. In this study, we applied label-free proteomics to discover plasma protein biomarkers in Thoroughbred geldings after administration with a long-acting form of recombinant human erythropoietin (rhEPO), methoxy polyethylene glycol epoetin beta, Mircera. Increased haematocrit, haemoglobin and red blood cell (RBC) levels were evidenced as early as 4 days post-administration in all three horses to varying extents. Tryptic peptides were obtained from plasma samples and analysed by nanoflow ultra-high-performance liquid chromatography-high-resolution tandem mass spectrometry (nano-UHPLC-HRMSMS) using data-independent acquisition. Differential protein abundance analysis has shortlisted seven protein biomarker candidates that showed significant changes specifically after Mircera administration in the treated but not in the control geldings, which comprised downregulation of two proteins, haptoglobin (HP) and haemopexin (HPX), and upregulation of five proteins, transferrin receptor 1 (TFR1), phospholipid transfer protein (PLTP), tenascin C (TNC), vascular cell adhesion molecule 1 (VCAM1) and galectin 3 binding protein (LGALS3BP). Multivariate analysis of plasma proteome has allowed the classification of control and treated samples. This is the first report on the discovery of plasma protein biomarkers of rhEPO administration to geldings. The results lay a foundation for applications of protein biomarkers for controlling the misuse of ESAs.