Pharmacokinetic study of osilodrostat and identification of mono-hydroxylated metabolite in equine plasma for the purpose of doping control.

RATIONALE: Osilodrostat is an inhibitor of 11-beta-hydroxylase (CYP11B) and is used for the treatment of Cushing's disease but also categorized as an anabolic agent. The use of osilodrostat is prohibited in horseracing and equestrian sports. To the best of our knowledge, this is the first metabolic study of osilodrostat in equine plasma. METHODS: Potential metabolites of osilodrostat were identified by differential analysis using data acquired from pre- and post-administration plasma samples after protein precipitation with liquid chromatography electrospray ionization high-resolution mass spectrometry (LC/ESI-HRMS). [Correction added on 27 January 2023, after first online publication: In the preceding sentence, "C-HRMS" was changed to "LC/ESI-HRMS" in this version.] For quantification of osilodrostat, a strong cation exchange solid-phase extraction was employed, and the extracts were analyzed using LC/ESI-triple quadrupole tandem mass spectrometry (LC/ESI-QqQ-MS/MS) to establish its elimination profile. Such extracts were further analyzed using LC/ESI-HRMS to investigate the detectability of osilodrostat and its identified mono-hydroxylated metabolite over a 2-week sampling period. RESULTS: Mono-hydroxylated osilodrostat was identified based on the differential analysis and mass spectrometric interpretations, and it was found to be the most abundant metabolite in plasma. Elimination profile of osilodrostat in plasma was successfully established over the 24-h post-administration period. Both osilodrostat and its mono-hydroxylated metabolite were detected up to the last sampling point at 2 weeks using HRMS, and osilodrostat could be confirmed up to 8-day post-administration with its reference material using HRMS as well. CONCLUSIONS: For doping control, screening of both the parent drug osilodrostat and its mono-hydroxylated metabolite in equine plasma would be recommended due to their extended detection windows of up to 2 weeks. Given the availability of reference material for potential confirmation in forensic samples, osilodrostat is considered the most appropriate monitoring target.

Epidemiology of jockey falls and injuries in flat and jump races in Japan (2003-2017).

Jockey safety is of paramount importance from the standpoint of welfare and public perception. Thus, an understanding of the epidemiology and associated risk factors is necessary to implement measures to reduce the jockey falls (JFs) and jokey injuries (JIs). This descriptive epidemiological study investigated the occurrence of JFs and JIs in 715,210 and 25,183 rides in flat and jump races, respectively, from 2003 to 2017. In flat races, the incidence rates of JFs and JIs were 1.4 and 0.6 per 1,000 rides, respectively. In jump races, they were 44.4 and 18.1 per 1,000 rides, respectively. In flat races, 56.8% of JFs at corners resulted in JIs. In jump races, the major causes of JFs and JIs were lost balance and hampered by a fallen horse at an obstacle. Our findings provide a basis to design a future study analyzing risk factors for JFs.

Prevalence of post-race exertional heat illness in Thoroughbred racehorses and climate conditions at racecourses in Japan.

Despite growing recognition of post-race exertional heat illness (EHI) in the horse racing industry, reports on its prevalence are limited. The purpose of this study was to investigate the prevalence of post-race EHI and climate conditions at racecourses in Japan. The overall prevalence of EHI from 1999 to 2018 was 0.04% (387 cases for 975,247 starters) in races operated by the Japan Racing Association (JRA). The yearly prevalence has been increasing, exceeding 0.07% in the last four years of the studied period. The overall prevalence in summer (May-September) was 0.086% (352 cases for 409,908 starters). The monthly prevalence varied among the 10 JRA racecourses, which are distributed from latitude 34 to 43°N, ranging from no cases to 0.459%. During summer, prevalence of post-race EHI was high when the mean monthly ambient temperature was high at a racecourse. To evaluate climate conditions, we investigated the wet-bulb globe temperature (WBGT, °C) from 9 AM to 5 PM on sunny race days in July and August of 2017 and 2018 at three racecourses with a high prevalence of EHI among the 10 racecourses. The durations of time during which WBGT was between 28 and 33°C at these three courses were 95, 94, and 65% of the minutes measured, respectively. This result indicated that most races on the sunny summer days were held when WBGT was between 28 and 33°C at the three racecourses. These findings could be useful in developing the appropriate countermeasures to be taken during hot weather at each of the studied racecourses.

Evaluating continuous blood coagulopathy in assessing the severity of acute colitis in Thoroughbred racehorses.

Although severe blood coagulopathy in horses with acute colitis causes multiple organ failure, which may be fatal, few studies have focused on the correlation between the fluctuations of coagulation parameters and severity of colitis. In this study, we evaluated the fluctuations of coagulation parameters in 14 Thoroughbred racehorses with acute colitis for 5 days from the day of hospitalization and compared them between 5 survivors and 9 non-survivors. Noteworthy features in the non-survivors were that antithrombin activity and fibrin degradation products continuously decreased and increased, respectively, for 4 days or more and that thrombin-antithrombin complexes increased in the last 2 days before death. Thus, these parameters should be continuously monitored to observe these fluctuations in assessing the severity of acute colitis.

Blood glucose is unlikely to be a prognostic biomarker in acute colitis with systemic inflammatory response syndrome in Thoroughbred racehorses.

Although hyperglycemia at admission with colic has been reported to have a poor prognosis, there is no report specifically about acute colitis with systemic inflammatory response syndrome (SIRS) in horses. In this study, we measured blood glucose (Glu), insulin (Ins), and cortisol (Cor) levels in 17 Thoroughbred racehorses diagnosed as having acute colitis with SIRS, and examined the relationship between time-dependent changes in Glu, Ins, and Cor and prognosis. Glu levels were high in 3 horses at admission, but thereafter no horses had persistently high Glu levels. There was no significant difference in Glu, Ins, and Cor levels within 72 hr between surviving and non-surviving horses. In conclusion, the Glu level is unlikely to be a useful prognostic biomarker in acute colitis with SIRS.

Signal changes in standing magnetic resonance imaging of osseous injury at the origin of the suspensory ligament in four Thoroughbred racehorses under tiludronic acid treatment.

Problems associated with the proximal metacarpal region, such as an osseous injury associated with tearing of Sharpey's fibers or an avulsion fracture of the origin of the suspensory ligament (OISL), are important causes of lameness in racehorses. In the present study, four Thoroughbred racehorses (age range, 2-4 years) were diagnosed as having forelimb OISL and assessed over time by using standing magnetic resonance imaging (sMRI). At the first sMRI examination, all horses had 3 characteristic findings, including low signal intensity within the trabecular bone of the third metacarpus on T1-weighted images, intermediate-to-high signal intensity surrounded by a hypointense rim on T2*-weighted images, and high signal intensity on fat-suppressed images. Following the sMRI examination, all horses received 50 mg of tiludronic acid by intravenous regional limb perfusion once weekly for 3 weeks. Attenuation of the high signal intensity on T2*-weighted and fat-suppressed images was observed on follow-up sMRI in 3 horses. Following rest and rehabilitation, these 3 horses successfully returned to racing. In contrast, the other horse that did not show attenuation of the high signal intensity failed to return to racing. To our knowledge, this is the first report of OISL in Thoroughbred racehorses assessed over time by sMRI under tiludronic acid treatment. Our findings support the use of sMRI for examining lameness originating from the proximal metacarpal region to refine the timing of returning to exercise based on follow-up examinations during the recuperation period.

Clinical usefulness of intravenous constant rate infusion of fentanyl and medetomidine under sevoflurane anesthesia in Thoroughbred racehorses undergoing internal fixation surgery.

A total of 20 racehorses with longitudinal fractures underwent internal fixation surgery under sevoflurane anesthesia combined with infusion of medetomidine (3 µg/kg/hr) alone (10 horses, group M) or medetomidine and fentanyl (7 µg/kg/hr) (10 horses, group FM). In group FM, the end-tidal sevoflurane concentration during surgery was maintained significantly lower than in group M (2.8-2.9% for group M vs. 2.2-2.6% for group FM, P<0.01). The mean arterial blood pressure was maintained over 70 mmHg using dobutamine infusion (group M, 0.36-0.54 µg/kg/min; group FM, 0.27-0.65 µg/kg/min), and the recovery qualities were clinically acceptable in both groups. In conclusion, co-administration of fentanyl and medetomidine by constant rate infusion may be a clinically useful intraoperative anesthetic adjunct for horses to reduce the requirement of sevoflurane when they undergo orthopedic surgery.

Characteristic findings of magnetic resonance imaging (MRI) and computed tomography (CT) for severe chronic laminitis in a Thoroughbred horse.

A Thoroughbred horse with severe chronic laminitis of both forelimbs was evaluated on the same day with magnetic resonance imaging (MRI) and computed tomography (CT). Both MRI and CT revealed loss of the dorsal aspect of the cortical bone of the 3rd phalanx and sclerosis. CT reflected the status of the horny layer and bone of the affected feet, while MRI depicted inflammation of the laminar corium, together with tendon edema. On the 3-dimensional CT venogram, vessels were visualized in both the right and left forelimbs, although there was a difference in the vasculature of the coronary plexus and circumflex vessels between the right and left forelimbs. A combination of both MRI and CT provides detailed information regarding pathological conditions.

Methicillin-resistant Staphylococcus aureus ulcerative keratitis in a Thoroughbred racehorse.

We report the first case of methicillin-resistant Staphylococcus aureus (MRSA) keratitis in a racehorse. A 5-year-old mare developed punctate keratitis after racing. The corneal ulcer continued to expand despite ophthalmic antimicrobial therapy. On day 6, a conjunctival graft surgery was performed. The mare was euthanized, following colitis and laminitis development on day 10. MRSA was isolated from the corneal swab taken at the time of euthanasia. Immunohistochemical analysis demonstrated gram-positive and anti-S. aureus monoclonal antibody-positive cocci infiltration of the corneal stroma; and a diagnosis of MRSA ulcerative keratitis was made. An ophthalmic antimicrobial against the isolated MRSA did not improve the ocular lesion. The MRSA strain was found to be staphylococcal cassette chromosome mec type II, a strain frequently isolated from humans in Japan.

Risk factors for jockey falls in Japanese Thoroughbred jump racing.

Jockey safety is an important subject from a welfare perspective and public perception. This is the first retrospective case-control study that aims to identify risk factors associated with jockey falls (JF) in Thoroughbred jump races held by the Japan Racing Association (JRA). JF in 17,459 maiden-class race starts at eight racecourses from 2003 to 2017 were retrospectively reviewed. Data were extracted from a database and official accident reports maintained by the JRA. Thirteen possible risk factors were evaluated using multivariable logistic regression to identify those that were significantly associated with JF. A total of 724 JF were recorded, with an incidence rate of 41.5 falls per 1,000 starts (95% CI: 38.6-44.5). Final model included stable, horse age, year, season, course, horse sex, horse experience, and jockey experience. No two-way interactions were observed. Six risk factors were significantly associated with JF: Year (2003-2007 or 2008-2012 > 2013-2017; P = .0011), season (spring, autumn, or winter > summer; P = .0006), course type (dual direction > single direction; P < .0001), horse sex (female > male or gelding; P = .0003), horse experience (inexperienced horse > experienced horse; P < .0001) and jockey experience (apprentice jockey > experienced jockey; P = .0332) significantly affected the odds of JF. In agreement with overseas reports, our results suggest that the occurrence of JF is multifactorial and associated with jockey- and horse-related factors as well as environmental factors. To safeguard the welfare of jockeys, implementation of measures according to identified risk factors is recommended.

Risk Factors for Jockey Falls in Japanese Thoroughbred Flat Racing.

Jockey safety is of paramount importance from welfare perspective and public perception. This retrospective case-control study aims to identify risk factors associated with jockey falls (JF) in flat races of Japan Racing Association (JRA). JF in 715,210 race starts by 74,328 horses at 10 racecourses from 2003 to 2017 were reviewed. Data were extracted from a database maintained by JRA and from official accident reports issued by race stewards. Seventeen possible risk factors were evaluated using multivariable logistic regression, to identify those significantly associated with JF. A total of 992 JF incidents were recorded, with an incidence rate of 1.39 falls per 1,000 starts (95% CI: 1.30-1.48). 6 risk factors were significantly associated with JF. Odds increased with horses that sustained catastrophic musculoskeletal injury (CMI) (OR: 203; CI: 169-241; P < 0.001). Increased odds were also associated with dirt track surfaces (OR: 1.99; CI: 1.74-2.29; P < 0.001), apprentice jockeys (OR: 1.43; CI: 1.21-1.68; P < 0.001), smaller track sizes (OR: 1.41; CI: 1.24-1.61; P < 0.001), larger fields (OR: 1.25; CI: 1.07-1.47; P = 0.005), and longer race distances (OR per 200 m: 1.05; CI: 1.01-1.09; P = 0.02). Since CMI was identified as a major contributing factor for JF, measures to minimize CMI may lead to improvement of jockey safety. The increased odds associated with apprentice jockeys may indicate the importance of jockey education and training. For jockey safety, proper staffing of medical professionals especially for races on dirt, smaller track, larger fields, and longer distances is recommended.

Presence of Antimicrobials in Postrace Samples in Japanese Thoroughbred Racing.

Ever since 'One Health' concept was introduced in early 2000s, judicious use of antimicrobials by veterinarians has become an issue of great concern. Recently, findings of anti-inflammatory effects in certain types of antimicrobials have raised a subject for discussion among racing authorities. Regulatory framework of antimicrobials in racing should be based on best interest of horse welfare and doping control perspective, but basic data on prevalence of antimicrobials are lacking. Analysis of 100 postrace urinary samples collected from 10 Japanese racecourses by targeting 21 antimicrobials using ultra performance liquid chromatography-tandem mass spectrometry resulted in detection of ceftiofur, cefalotin, cefalotin metabolite, dihydrostreptomycin, gentamicin, kanamycin, and oxytetracycline. Detection of antimicrobials critically important for resistance in human medicine was limited to a single sample. Oxytetracycline, which is known to possess anti-inflammatory effects, was detected in three samples. This may suggest the need for establishing a regulatory framework from doping control perspective and further studies to clarify pharmacologically relevant concentration of antimicrobials with such properties.

Bone marrow oedema-type signal in the proximal phalanx of Thoroughbred racehorses.

This study focused on 8 Thoroughbred racehorses showing bone marrow oedema-type signal in the proximal sagittal groove of the proximal phalanx, with the aim of understanding its clinical significance. Standing magnetic resonance imaging played an important role in assessing osseous abnormalities that were not radiographically identifiable. Further, a histopathological result from one of the cases showed there was oedema surrounding adipose tissues with increase in density of trabecular scaffolding. This may indicate presence of osseous injury within the area of decreased elasticity due to subchondral bone modeling. This study suggests that detection of osseous abnormality based on bone marrow oedema-type signal, and application of appropriate care following injury would contribute to prevent deterioration of stress-related fractures of the proximal phalanx.

Standing magnetic resonance imaging of distal phalanx fractures in 6 cases of Thoroughbred racehorse.

Six Thoroughbred racehorses with palmar process fractures of the distal phalanx were evaluated with standing magnetic resonance imaging (sMRI). In all the cases, the fractures were detectable on T1-weighted images and fat-suppressed images. Furthermore, multi-planar reconstruction images were useful for assessing the articular involvement of the fractures. Follow-up sMRI was obtainable in 3 cases, which revealed that the area of high signal intensity on fat-suppressed images decreased over time as symptoms improved. Our findings support the use of sMRI for the detailed evaluation of distal phalanx fractures including their articular involvement and the healing process.

Pharmacokinetics and pharmacodynamics of olopatadine following administration via nasogastric tube to healthy horses.

OBJECTIVE: To investigate the pharmacokinetics and antihistaminic effects (pharmacodynamics) of olopatadine in a small population of healthy horses after administration via nasogastric tube. ANIMALS: 4 healthy adult Thoroughbreds. PROCEDURES: Olopatadine (0.1 mg/kg, once) was administered via nasogastric tube. Blood samples were collected at predetermined time points for pharmacokinetic analyses of the drug in plasma. Olopatadine effects were investigated by measurement of cutaneous wheals induced by ID histamine injection (0.1 mL [10 µg]/injection) at predetermined time points. Inhibition effect ratios were calculated on the basis of measured wheal size (area) after versus before olopatadine administration. RESULTS: Mean ± SD maximum plasma olopatadine concentration was 48.8 ± 11.0 ng/mL approximately 1.5 hours after administration. Median terminal half-life was 6.11 hours. Mean ± SD maximal effect was 88.2 ± 4.9% inhibition approximately 3.5 hours after drug delivery, and the inhibition effect remained > 80% for 12.5 hours after treatment. No signs of adverse clinical effects were observed. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested olopatadine may have a strong, long-term inhibitory effect against histamine-induced wheals in the skin of horses. Clinical research with a larger number of horses is warranted.

Use of standing low-field magnetic resonance imaging to assess oblique distal sesamoidean ligament desmitis in three Thoroughbred racehorses.

Desmitis of the oblique distal sesamoidean ligaments (ODSL) is caused by hyperextension of the metacarpophalangeal/metatarsophalangeal joint and has been described as a significant cause of lameness in racehorses. In this study, three Thoroughbred racehorses (age range: 3-6 years) were diagnosed with desmitis of the forelimb ODSL using standing low-field magnetic resonance imaging (sMRI). Radiography and ultrasonography were inconclusive with regard to a definitive diagnosis. For all horses, the sMRI characteristics included increased signal intensity within the medial ODSL on T1-weighted gradient echo, T2-weighted fast spin echo and short tau inversion recovery fast spin echo images, which use a fat suppression technique. Effusion of the digital flexor tendon sheath was also clearly visible on sMRI. Following rest and controlled exercise for roughly 3 months, two horses successfully returned to racing within 5 months. Our findings support the use of sMRI for diagnosing ODSL injuries in Thoroughbred racehorses.

Pharmacokinetics and pharmacodynamics of d-chlorpheniramine following intravenous and oral administration in healthy Thoroughbred horses.

The pharmacokinetics of d-chlorpheniramine (CPM), a histamine H1-receptor antagonist, and its ability to inhibit of histamine-induced cutaneous wheal formation, were studied in healthy Thoroughbred horses (n=5). Following an intravenous (IV) dose of 0.5mg/kg bodyweight (BW), plasma drug disposition was very rapid, with the mean terminal half-life and total body clearance calculated as 2.7h and 0.7 L/h/kg, respectively. The observed maximal inhibition of wheal formation following IV doses of 0.1 and 0.5mg/kg BW were 37.8% and 60.6% at 0.5h, respectively. Oral administration of CPM (0.5mg/kg BW) resulted in a bioavailability of 38%, which induced a peak plasma drug concentration at 1h and a maximal inhibition of wheal formation (39%) at 2h. A pharmacokinetic/pharmacodynamic link model showed that CPM in horses has lower efficacy, much lower potency and slightly lower sensitivity than other reported antihistamines. These results indicated that CPM should be administered at frequent intervals or at large dose rates to maintain therapeutic concentrations in horses.