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Topic:Urine Analysis
Urine analysis in horses involves the examination of urine to assess the health and physiological status of the animal. This diagnostic tool provides insights into the renal function, hydration status, and potential presence of systemic diseases. Key parameters evaluated in equine urine analysis include pH, specific gravity, protein levels, glucose, ketones, and the presence of blood or sediment. Variations in these parameters can indicate metabolic disorders, infections, or other health issues. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, findings, and clinical implications of urine analysis in equine veterinary practice.
Nephrogenic diabetes insipidus in a 14-year-old gelding. A 14-year-old Cleveland Bay cross gelding was presented with severe urinary incontinence that had been present for 1 year, and chronic polydipsia and polyuria over 4 years. Water intake had been recorded as 240 L over a 24-hour period. Results: The horse had marked urinary incontinence and polyuria and polydipsia. The urine was markedly hyposthenuric, but no abnormalities on urinalysis were detected. There were no other abnormal clinical or neurological signs. Haematological and serum biochemical examinations showed no abnormalities and ultrasonographic and endoscopic examination of the urinar...
Metabolic studies of 1-testosterone in horses. 1-Testosterone (17β-hydroxy-5α-androst-1-en-3-one), a synthetic anabolic steroid, has been described as one of the most effective muscle-building supplements currently on the market. It has an anabolic potency of 200 as compared to 26 for testosterone. Apart from its abuse in human sports, it can also be a doping agent in racehorses. Metabolic studies on 1-testosterone have only been reported for human in the early seventies, whereas little is known about its metabolic fate in horses. This paper describes the studies of in vitro and in vivo metabolism of 1-testosterone in horses, with the ai...
An alternative dietary strategy to make weight while improving mood, decreasing body fat, and not dehydrating: a case study of a professional jockey. Professional jockeys are unique among weight-making athletes, as they are often required to make weight daily and, in many cases, all year-round. Common methods employed by jockeys include dehydration, severe calorie restriction, and sporadic eating, all of which have adverse health effects. In contrast, this article outlines a structured diet and exercise plan, employed by a 22-yr-old professional National Hunt jockey in an attempt to reduce weight from 70.3 to 62.6 kg, that does not rely on any of the aforementioned techniques. Before the intervention, the client's typical daily energy intak...
p-Cresol: a sex pheromone component identified from the estrous urine of mares. Previously it was shown that m- and p-cresols in the urine of mares exhibits a temporally reproducible pattern that is dependent on ovarian activity and, thus, provides information about the timing of ovulation. New behavioral data demonstrate 1) that stallions spend significantly more time sniffing p-cresol as compared to o-, and m-cresols, and, 2) that the extent of stallions' erections differ significantly in response to different types of samples. The lowest erection level was recorded for the pure-water control, a moderate erection level was elicited by the urine of diestrous mares, and t...
Structural elucidation of phase I and II metabolites of bupivacaine in horse urine and fungi of the Cunninghamella species using liquid chromatography/multi-stage mass spectrometry. Bupivacaine is a local anaesthetic prohibited in equine sports. It is highly metabolized in the horse but a thorough description of its metabolite profile is lacking. An administration study should find appropriate analytical targets for doping control. Furthermore, knowledge of an in vitro system for production of metabolites would be beneficial. Methods: Marcain® (bupivacaine hydrochloride) was administered subcutaneously to a horse and urine samples were collected. In vitro metabolic systems consisting of the fungi Cunninghamella elegans and Cunninghamella blakesleeana were incubated with ...
Evaluation of a modification of the McKinnon technique to correct urine pooling in mares. The urethral fold of 30 mares was split transversely into dorsal and ventral shelves, and the ventral shelf was used to help create a urethral extension. The dorsal shelf was stretched caudally and sutured to the roof of the extension so that it covered at least the cranial half of the extension. For 20 mares, a relaxing, vaginal incision was created cranial to the external urethral orifice to enable the dorsal shelf to be retracted further caudally. Ten of the 30 mares (33.3 per cent) developed a defect, but none developed a defect in that portion covered by the dorsal shelf of the urethral f...
Acepromazine pharmacokinetics: a forensic perspective. Acepromazine (ACP) is a useful therapeutic drug, but is a prohibited substance in competition horses. The illicit use of ACP is difficult to detect due to its rapid metabolism, so this study investigated the ACP metabolite 2-(1-hydroxyethyl)promazine sulphoxide (HEPS) as a potential forensic marker. Acepromazine maleate, equivalent to 30mg of ACP, was given IV to 12 racing-bred geldings. Blood and urine were collected for 7days post-administration and analysed for ACP and HEPS by liquid chromatography-mass spectrometry (LC-MS). Acepromazine was quantifiable in plasma for up to 3h with little r...
Detection of fluticasone propionate in horse plasma and urine following inhaled administration. Fluticasone propionate (FP) is an anti-inflammatory agent with topical and inhaled applications commonly used in the treatment of asthma in steroid-dependent individuals. The drug is used in racehorses to treat Inflammatory Airway Disease; this work was performed in order to advise on its use and detect potential misuse close to racing. Methods were developed for the extraction and analysis of FP from horse plasma and a carboxylic acid metabolite (FP-17βCOOH) from horse urine. The methods utilize ultra high performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) in ...
Detection of recombinant human EPO administered to horses using MAIIA lateral flow isoform test. Doping of horses with recombinant human erythropoietin (rHuEPO) to illegally enhance their endurance capacity in horseracing has been reported during the last years. This leads to increased blood viscosity which can result in sudden death and is of concern for the horse welfare. Additionally, the horse can start production of rHuEPO antibodies, which cross-reacts with endogenous equine EPO and can lead to severe anaemia and even death. In this study, a novel micro-chromatographic method, EPO WGA MAIIA, has been tested for the capability in plasma and urine samples to detect administration of e...
Comparison of the effects of xylazine bolus versus medetomidine constant rate infusion on the stress response, urine production, and anesthetic recovery characteristics in horses anesthetized with isoflurane. To compare the effect of xylazine bolus versus medetomidine constant rate infusion (MCRI) on serum cortisol and glucose concentrations, urine production, and anesthetic recovery characteristics in dorsally recumbent, spontaneously breathing, isoflurane-anesthetized horses. Methods: Prospective, randomized crossover study. Methods: 10 healthy Standardbreds. Methods: Horses were premedicated with xylazine or medetomidine IV. Anesthesia was induced with diazepam and ketamine and maintained with isoflurane for 150 minutes. For the xylazine treatment, end-tidal isoflurane concentration was maintain...
A new analytical method based on anti-EPO monolith column and LC-FAIMS-MS/MS for the detection of rHuEPOs in horse plasma and urine samples. Recombinant human erythropoietin (rHuEPO) is a 30-34 kDa glycoprotein banned by the racing authorities. For some years this molecule has been detected in race horses in USA and in Europe, and even in racing camels. Although direct methods to differentiate horse endogenous EPO and rHuEPO have been developed either by LC-MS/MS or by isoelectric focusing (IEF) with double-blotting, the short confirmation time of such prohibited hormone in plasma remains a problem for horseracing doping control laboratories. In order to improve the rHuEPOs confirmation process in horse plasma or urine in terms of ...
The role of horses in the transmission of leptospirosis in an urban tropical area. The objective of this study was to demonstrate the presence of leptospires in equine urine, as evidence for a potential role of horses in transmission of this organism. Thoroughbred horses (aged 2-5 years, n = 276) from Rio de Janeiro, Brazil, were studied. After a severe storm, the premises of the animals remained flooded for 72 h. Blood samples for serology were collected on days 20 and 35 (day of storm = day 0). On day 20, 132 (47·8%) horses were seroreactive (titre ≥200) and, of these, 23 (31·0%) had increased antibody titres on day 35. Furthermore, 34 urine samples (for PCR and cultur...
Investigation of the presence of endogenous prednisolone in equine urine by high-performance liquid chromatography mass spectrometry and high-resolution mass spectrometry. After the detection of low concentrations of prednisolone in racehorse urine samples collected at Italian racetracks, a study was initiated to investigate the accuracy of the analytical protocol used and the possible endogenous origin of detected prednisolone. Methods: Multiple reaction monitoring (MRM) MS(2) acquisition with a triple quadrupole (n = 780) and full scan MS(2) and MS(3) (n = 180) acquisition with a linear ion trap were checked. As a further confirmation, ten urine samples were analysed by high-resolution mass spectrometry (HRMS). Results: The study showed the difficulty of ident...
Detection of myo-inositol trispyrophosphate in equine urine and plasma by hydrophillic interaction chromatography-tandem mass spectrometry. Myo-inositol trispyrophosphate (ITPP) is a new drug capable of increasing the amount of oxygen in hypoxic tissues. Studies have shown that administration of ITPP increases the maximal exercise capacity in normal mice as well as mice with severe heart failure. The properties of ITPP make it an ideal candidate as a doping agent to enhance performance in racehorses. While there have been speculations in the horseracing industry that the covert use of ITPP is already widespread, no reported method exists for the detection of ITPP in equine biological samples. ITPP is a difficult-to-detect drug due...
Pharmacokinetics and pharmacodynamics of butorphanol following intravenous administration to the horse. Butorphanol is a narcotic analgesic commonly used in horses. Currently, any detectable concentration of butorphanol in biological samples collected from performance horses is considered a violation. The primary goal of the study reported here was to update the pharmacokinetics of butorphanol following intravenous administration, utilizing a highly sensitive liquid chromatography-mass spectrometry (LC-MS) assay that is currently employed in many drug-testing laboratories. An additional objective was to characterize behavioral and cardiac effects following administration of butorphanol. Ten exer...
Regulatory control of glycopyrrolate in performance horses using validated UHPLC/MS-MS methods. We describe a validated, rapid, sensitive, and specific UHPLC-MS/MS method to detect and quantify glycopyrrolate in 0.5 mL of horse urine. Further, we investigated the elimination of glycopyrrolate in urine after both intravenous and oral administration of clinically relevant doses to Thoroughbred horses. Quantification was performed by weighted, linear regression analysis using a deuterated analogue of glycopyrrolate as internal standard (IS). The method was characterized by a linear range of 5-2500 pg/mL, a lower limit of quantification of 5 pg/mL and a limit of detection of 1 pg/mL. The int...
Abnormal reproductive patterns in Przewalski’s mares are associated with a loss in gene diversity. The ex situ population of the Przewalski's horse (Equus ferus przewalskii) is not self-sustaining (20% foaling rate), and the demography is skewed toward aging individuals with low gene diversity. We designed the present study to gain a better understanding of the reproductive biology of the Przewalski's mare and to determine whether age and gene diversity influenced reproductive function. Urine samples were collected 3-7 days/wk from 19 mares from May to September, and ultrasound examinations of follicular structures were performed 3 days/wk for 5 wk from May through July in nine individuals....
Rapid screening of anabolic steroids in horse urine with ultra-high-performance liquid chromatography/tandem mass spectrometry after chemical derivatisation. Liquid chromatography/mass spectrometry (LC/MS) has been successfully applied to the detection of anabolic steroids in biological samples. However, the sensitive detection of saturated hydroxysteroids, such as androstanediols, by electrospray ionisation (ESI) is difficult because of their poor ability to ionise. In view of this, chemical derivatisation has been used to enhance the detection sensitivity of hydroxysteroids by LC/MS. This paper describes the development of a sensitive ultra-high-performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS) method for the screening of a...
Case report: squamous cell carcinoma of the urinary bladder in a horse. A 14-year-old Hanoverian gelding was evaluated after a 10-week history of occasional straining during urination and dark red urine production after exercise. Physical examination findings were within normal limits. After 2 weeks of clinical signs, ultrasonography had found no abnormalities in the kidneys or bladder. A freely voided urine sample revealed an elevated protein concentration (500 mg/dL; reference range: <100 mg/dL) and few red blood cells. Vitamin C supplementation was suggested to acidify the urine, as was provision of a salt block to encourage water intake.
p- and m-Cresols emitted from estrous urine are reliable volatile chemical markers of ovulation in mares. Urine samples from 14 mares, belonging to five breeds, were collected at estrus and diestrus to search for estrous specific volatile compounds which could be used to determine ovulation time. Around 150 volatiles were collected from urine head-space samples by solid phase micro extraction technique, and analyses were conducted by gas chromatography-mass spectrometry methods. Comparison of chromatographic profiles of volatile substances revealed that concentrations of m- and p-cresols were significantly greater during estrus while diethylphthalate was more abundant at diestrus. Monitoring of m-...
The levels of zearalenone and its metabolites in plasma, urine and faeces of horses fed with naturally, Fusarium toxin-contaminated oats. Concentration profile of zearalenone (ZON) and its metabolites in plasma, urine and faeces samples of horses fed with Fusarium toxin-contaminated oats is described. In plasma, β-zearalenol (β-ZOL) was detected at high levels on day 10 of the study (3.21-6.24 μg/l). β-Zearalenol and α-zearalenol were the major metabolites in urine. Zearalenone, α-ZOL and β-ZOL were predominantly found in faeces. Zearalanone could also be detected in urine (1.34-5.79 μg/l) and faeces (1 μg/kg). The degree of glucuronidation was established in all sample types, approximately 100% in urine and plasm...
Detection of singly- and doubly-charged quaternary ammonium drugs in equine urine by liquid chromatography/tandem mass spectrometry. Quaternary ammonium drugs (QADs) are anticholinergic agents some of which are known to have been abused or misused in equine sports. A recent review of literature shows that the screening methods reported thus far for QADs mainly cover singly-charged QADs. Doubly-charged QADs are extremely polar substances which are difficult to be extracted and poorly retained on reversed-phase columns. It would be ideal if a comprehensive method can be developed which can detect both singly- and doubly-charged QADs. This paper describes an efficient liquid chromatography/tandem mass spectrometry (LC/MS/MS) m...
Ex vivo spontaneous generation of 19-norandrostenedione and nandrolone detected in equine plasma and urine. 19-Norandrostenedione (NAED) and nandrolone are anabolic-androgenic steroids (AASs). Nandrolone was regarded solely as a synthetic AAS until the 1980s when trace concentrations of apparently endogenous nandrolone were detected in urine samples obtained from intact male horses (stallions). Since then, its endogenous origin has been reported in boars and bulls; endogenous NAED and nandrolone have been identified in plasma and urine samples collected from stallions. More recently, however, it was suggested that NAED and nandrolone detected in urine samples from stallions are primarily artifacts d...
Quantitative analysis of a quaternary ammonium drug: ipratropium bromide by LC/ESI-MS(n) in horse plasma and urine. A quantitative method, using LC/ESI-MS(n) with a quadrupole linear ion trap mass analyzer, has been developed for the analysis of ipratropium cation in horse plasma and urine. The method applies solid-phase extraction with WCX cartridges for plasma and MM2 cartridges for urine, prior to analysis by LC/ESI-MS(n). The efficiency of extraction combined with the sensitivity and the selectivity of MS(n) allows for the quantification of ipratropium cation at picogram per milliliter levels. The analytical capabilities of the method have been successfully checked by the quantitative analysis of ipratr...
Pharmacokinetics and pharmacodynamics of detomidine following sublingual administration to horses. To characterize pharmacokinetics and pharmacodynamics of detomidine gel administered sublingually in accordance with label instructions to establish appropriate withdrawal guidelines for horses before competition. Methods: 12 adult racehorses. Methods: Horses received a single sublingual administration of 0.04 mg of detomidine/kg. Blood samples were collected before and up to 72 hours after drug administration. Urine samples were collected for 5 days after detomidine administration. Plasma and urine samples were analyzed via liquid chromatography-mass spectrometry, and resulting data were anal...
Horse metabolism and the photocatalytic process as a tool to identify metabolic products formed from dopant substances: the case of sildenafil. Two horses were treated with sildenafil, and its metabolic products were sought in both urine and plasma samples. Prior to this, a simulative laboratory study had been done using a photocatalytic process, to identify all possible main and secondary transformation products, in a clean matrix; these were then sought in the biological samples. The transformation of sildenafil and the formation of intermediate products were evaluated adopting titanium dioxide as photocatalyst. Several products were formed and characterized using the HPLC/HRMS(n) technique. The main intermediates identified in thes...
Liquid chromatography electrospray ionization tandem mass spectrometry for the detection of mesocarb abuse in horse doping. A method is described for the determination of mesocarb abuse in equestrian sport by combining gradient liquid chromatography and electrospray ionization tandem mass spectrometry. Mesocarb was administrated orally to two horses at a dose of 50 µg/kg. Urine samples were collected up to 120 h post administration. Hydrolyzed and conjugated urine fractions were handled using liquid-liquid extraction (LLE). The identity of the parent drug and metabolites was confirmed using liquid chromatography combined with tandem mass spectrometry (MS/MS). Mesocarb and seven metabolites were detected in horse...
A combination of single-drop microextraction and open tubular capillary electrochromatography with carbon nanotubes as stationary phase for the determination of low concentration of illicit drugs in horse urine. In this study we developed an interesting alternative to HPLC-mass spectrometry for the quantification of seven important drugs of abuse in racehorses. The procedure proposed in this work is a combination of single-drop microextraction (SDME) and an open tubular capillary electrochromatography (OT-CEC) using multi-wall carbon nanotubes (MWCTs) immobilized into a fused-silica capillary as a stationary phase. The SDME showed to be a powerful tool for extraction/preconcentration of the seven drugs analyzed in the study, showing an enrichment factor between 38- and 102-fold depending on the drug. ...
In vitro metabolism of tiletamine, zolazepam and nonbenzodiazepine sedatives: Identification of target metabolites for equine doping control. Within horseracing, the detection of prohibited substance doping often requires urine analysis; hence, it is necessary to understand the metabolism of the drugs in question. Here, the previously unknown equine metabolism of eight sedatives is reported in order to provide information on target metabolites for use in doping control. Phase I metabolite information was provided by incubation with equine liver S9 fraction. In vitro techniques were chosen in order to reduce the ethical and financial issues surrounding the study of so many compounds, none of which are licensed for use in horses in th...
Detection of various performance enhancing substances in specimens collected from race horses in Illinois: a five-year experience. In order to protect the integrity of horse racing in Illinois, a complex testing of urine and blood specimens collected post-race from winning and special designation horses is continuously conducted. The initial screening by immunoassays was followed by the confirmation on presumptive positive samples. Instrumental screening was also conducted. Perimortem and postmortem specimens and special exhibits (syringes, needles, etc.) were also analyzed. The administration of alkalinizing agents was detected by measuring the total plasma carbon dioxide concentration. The laboratory analyzed specimens ...
