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.
Tang FP, Leung GN, Wan TS.A capillary electrophoresis-mass spectrometry (CE-MS) method for the analysis of quaternary ammonium drugs in equine urine was developed. Quaternary ammonium drugs were first extracted from equine urine by ion-pair extraction and then analysed by CE-MS in the positive electrospray ionization (ESI) mode. Within 12 min, eight quaternary ammonium drugs, each at 1 ng/mL in horse urine, could be detected. The confirmation of these drugs in urine samples was achieved by capillary electrophoresis tandem mass spectrometry (CE-MS/MS). A direct comparison of this method was made with existing liquid chr...
Haines GR, Brown MP, Gronwall RR, Merritt KA, Baltzley LK.Pharmacokinetics and distribution of orbifloxacin into body fluids and endometrium was studied in 6 mares after intragastric (IG) administration at a single dose rate of 7.5 mg/kg body weight. Orbifloxacin concentrations were serially measured in serum, synovial fluid, peritoneal fluid, urine, cerebrospinal fluid, and endometrial tissues over 24 hours. Minimum inhibitory concentrations of orbifloxacin were determined for 120 equine pathogens over an 11-month period. The mean peak serum concentration (Cmax) was 2.41+/-0.30 microg/mL at 1.5 hours after administration and decreased to 0.17+/-0.01...
Harkins JD, Karpiesiuk W, Lehner A, Woods WE, Dirikolu L, Carter WG, Boyles J, Tobin T.This report evaluates the pharmacological responses, urinary detection and mass spectral confirmation of ropivacaine in horses. Ropivacaine, a potent local anesthetic (LA) recently introduced in human medicine, has an estimated highest no-effect dose (HNED) of about 0.4 mg/site as determined in our abaxial sesamoid block model. Apparent ropivacaine equivalents were detectable by ELISA screening using a mepivacaine ELISA test after administration of clinically effective doses. Mass spectral examination of postadministration urine samples showed no detectable parent ropivacaine, but a compound i...
Queiroz-Neto A, Zamur G, Carregaro AB, Mataqueiro MI, Salvadori MC, Azevedo CP, Harkins JD, Tobin T.Caffeine is the legal stimulant consumed most extensively by the human world population and may be found eventually in the urine and/or blood of race horses. The fact that caffeine is in foods led us to determine the highest no-effect dose (HNED) of caffeine on the spontaneous locomotor activity of horses and then to quantify this substance in urine until it disappeared. We built two behavioural stalls equipped with juxtaposed photoelectric sensors that emit infrared beams that divide the stall into nine sectors in a 'tic-tac-toe' fashion. Each time a beam was interrupted by a leg of the horse...
Toribio RE, Kohn CW, Chew DJ, Sams RA, Rosol TJ.To evaluate calcium balance and parathyroid gland function in healthy horses and horses with enterocolitis and compare results of an immunochemiluminometric assay (ICMA) with those of an immunoradiometric assay (IRMA) for determination of serum intact parathyroid hormone (PTH) concentrations in horses. Methods: 64 horses with enterocolitis and 62 healthy horses. Methods: Blood and urine samples were collected for determination of serum total calcium, ionized calcium (Ca2+) and magnesium (Mg2+), phosphorus, BUN, total protein, creatinine, albumin, and PTH concentrations, venous blood gases, and...
Soma LR, Uboh CE, Guan F, Birks EK, Teleis DC, Rudy JA, Tsang DS, Watson AO.To determine pharmacokinetics and excretion of phenytoin in horses. Methods: 6 adult horses. Methods: Using a crossover design, phenytoin was administered (8.8 mg/kg of body weight, IV and PO) to 6 horses to determine bioavailability (F). Phenytoin also was administered orally twice daily for 5 days to those same 6 horses to determine steady-state concentrations and excretion patterns. Blood and urine samples were collected for analysis. Results: Mean (+/- SD) elimination half-life following a single IV or PO administration was 12.6+/-2.8 and 13.9+/-6.3 hours, respectively, and was 11.2+/-4.0 ...
McKinney AR, Ridley DD, Suann CJ.After oral administration to a thoroughbred gelding, the anabolic steroid norethandrolone was converted into a complex mixture of oxygenated metabolites. These metabolites were extracted from the urine, deconjugated by methanolysis and converted to their O-methyloxime trimethylsilyl derivatives. Gas chromatographic/mass spectrometric analysis indicated the major metabolites to be 19-norpregnane-3,16,17-triols, 19-norpregnane-3,17,20-triols and 3,17-dihydroxy-19-norpregnan-21-oic acids. Some minor metabolites were also detected.
Schott HC, Patterson KS, Eberhart SW.To determine whether administration of glycerol-containing solutions induces a state of transient hyperhydration in resting euhydrated horses, changes in plasma and urine constituents were measured in four horses for 1 h before and 5 h after nasogastric administration of each of four treatments (Experiment 1). Treatments were applied in a randomized fashion and included: (1) 1.0 g.kg(-)(1)glycerol in 8 L of water (G); (2) 8 L of water (W); (3) 8 L of 0.9% NaCl solution (S); and (4) 1.0 g.kg(-)(1)glycerol in 8 L of 0.9% NaCl solution (GS). In a subsequent study, voluntary water intake was measu...
Brinsko SP.Retrograde ejaculation was diagnosed in a 10-year-old Arabian stallion. Despite behavioral signs consistent with ejaculation, the collection receptacle of an artificial vagina remained devoid of semen on numerous occasions. Catheterization of the urinary bladder yielded large numbers of spermatozoa, even when an ejaculate was obtained, whereas low numbers (< 1 X 10(6)/ml) of spermatozoa are found in the bladder of clinically normal stallions after ejaculation. Endoscopic examination of the urethra, seminal colliculus, and bladder failed to reveal abnormalities. Medical treatment with imipra...
Harkins JD, Robinson NE, Woods WE, Lehner AF, Smith MD, Gates RS, Fisher M, Tobin T.Clenbuterol, a beta2 agonist/antagonist, is the only bronchodilator approved by the US Food and Drug Administration for use in horses. The Association of Racing Commissioners International classifies clenbuterol as a class 3 agent, and, as such, its identification in post-race samples may lead to sanctions. Anecdotal reports suggest that clenbuterol may have been administered by intratracheal (IT) injection to obtain beneficial effects and avoid post-race detection. The objectives of this study were (1) to measure the pharmacological efficacy of IT dose of clenbuterol and (2) to determine the ...
Guan F, Uboh CE, Soma LR, Birks EK, Teleis D, Rudy JA, Watson AO, Tsang DS.A reliable and sensitive method for the extraction and quantification of phenytoin (5,5'-diphenylhydantoin), its major metabolite, 5-(p-hydroxyphenyl)-5-phenylhydantoin (p-HPPH) and minor metabolite, 5-(m-hydroxyphenyl)-5-phenylhydantoin (m-HPPH) in horse urine and plasma is described. The method involves the use of solid-phase extraction (SPE), liquid-liquid extraction (LLE), enzyme hydrolysis (EH) and high-performance liquid chromatography (HPLC). The minor metabolite, 5-(m-hydroxyphenyl)-5-phenylhydantoin (m-HPPH) was not present in a reliably quantifiable concentration in all samples. The ...
Marshall DE, Mortishire-Smith RJ, Houghton E, Gower DB.Oestradiene-3,17-diol and oestratriene-3,17-diol (or the diol of Heard's ketone (3-hydroxy-5(10),6,8-oestratriene-17-one) have been extracted on a large scale from pooled urines and allantoic fluid obtained from pregnant mares. Initial purification was achieved using column chromatography, and further purification by high performance liquid chromatography or silver nitrate (argentation) thin layer chromatography. The steroids were characterised using gas chromatography-mass spectrometry. Positions of the double bonds in ring B of oestradienediol were deduced on the basis of results of ultravio...
Harkins JD, Karpiesiuk W, Tobin T, Dirikolu L, Lehner AF.Ropivacaine is a local anesthetic that has a high potential for abuse in racing horses. It can be recovered from urine collected after administration as a hydroxylated metabolite following beta-glucuronidase treatment of the urine. Based on these findings, it has been inferred that ropivacaine is present in equine urine as a glucuronide metabolite; however, these metabolites have never been directly identified. Using ESI+/MS/MS, the presence of a [M+H]+ molecular ion of m/z 467 was demonstrated in urine corresponding to the calculated mass of a hydroxyropivacaine glucuronide +1. The abundance ...
Lehner AF, Almeida P, Jacobs J, Harkins JD, Karpiesiuk W, Woods WE, Dirikolu L, Bosken JM, Carter WG, Boyles J, Holtz C, Heller T, Nattrass C....Remifentanil (4-methoxycarbonyl-4-[(1-oxopropyl)phenylamino]-1-piperidinepropionic acid methyl ester) is a mu-opioid receptor agonist with considerable abuse potential in racing horses. The identification of its major equine urinary metabolite, 4-methoxycarbonyl-4-[(1-oxopropyl)phenylamino]-1-piperidinepropionic+ ++ acid, an ester hydrolysis product of remifentanil is reported. Administration of remifentanil HCl (5 mg, intravenous) produced clear-cut locomotor responses, establishing the clinical efficacy of this dose. ELISA analysis of postadministration urine samples readily detected fentany...
Uboh CE, Soma LR, Luo Y, McNamara E, Fennell MA, May L, Teleis DC, Rudy JA, Watson AO.To compare the pharmacokinetics of penicillin G and procaine in racehorses following i.m. administration of penicillin G procaine (PGP) with pharmacokinetics following i.m. administration of penicillin G potassium and procaine hydrochloride (PH). Methods: 6 healthy adult mares. Methods: Horses were treated with PGP (22,000 units of penicillin G/kg of body weight, i.m.) and with penicillin G potassium (22,000 U/kg, i.m.) and PH (1.55 mg/kg, i.m.). A minimum of 3 weeks was allowed to elapse between drug treatments. Plasma and urine penicillin G and procaine concentrations were measured by use of...
Bryant JE, Brown MP, Gronwall RR, Merritt KA.The objectives of this study were to determine the pharmacokinetics and tissue concentrations of doxycycline after repeated intragastric administration, and to determine the minimum inhibitory concentrations (MIC) for equine pathogenic bacteria. In experiment 1, 2 mares received a single intragastric dose of doxycycline hyclate (3 mg/kg bwt). Mean peak serum concentration was 0.22 microg/ml 1 h postadministration. In experiment 2, 5 doses of doxycycline hyclate (10 mg/kg bwt), dissolved in water, were administered to each of 6 mares via nasogastric tube at 12 h intervals. The mean +/- s.e. pea...
Ribeiro Neto LM, Salvadori MC, Spinosa HS.As hydrocortisone is an endogenous substance, it is first necessary to establish its normal concentrations so as to be able to control its use in racing animals. This study was designed to establish the hydrocortisone concentrations in post-race urine samples of horses racing in Brazil and also to evaluate the results in relation to the international threshold set for this drug. Urine samples were analysed by HPLC-UV. The results were evaluated according to the concentration range as well as sex and time of sample collection (afternoon or evening races). The results showed a high degree of var...
Bosken JM, Lehner AF, Hunsucker A, Harkins JD, Woods WE, Karpiesiuk W, Carter WG, Boyles J, Fisher M, Tobin T.Isoxsuprine is routinely recovered from enzymatically-hydrolyzed, post-administration urine samples as parent isoxsuprine in equine forensic science. However, the specific identity of the material in horse urine from which isoxsuprine is recovered has never been established, although it has long been assumed to be a glucuronide conjugate (or conjugates) of isoxsuprine. Using ESI/MS/MS positive mode as an analytical tool, urine samples collected 4-8 h after isoxsuprine administration yielded a major peak at m/z 554 that was absent from control samples and resisted fragmentation to daughter ions...
Stanley SM.The metabolism and urinary excretion of a 100 mg dose of the non-sedating anxiolytic drug buspirone was examined using high-performance liquid chromatography/electrospray ionization mass spectrometry in the positive ion mode. In addition to a significant proportion of unchanged buspirone we were able to detect three major metabolite classes. These were identified as monohydroxy, dihydroxy and dihydroxymethoxy products. Detection of the metabolites and the parent drug was possible in all the urine samples collected (1-12 h) post-administration.
Törneke K, Larsson CI, Appelgren LE.Isoxsuprine is used in veterinary medicine as a vasodilating agent. The drug has been detected in the urine of horses up to 6 weeks after the cessation of administration. In the present study, the distribution pattern of 3H-isoxsuprine was investigated using whole body autoradiography in mice to find a possible site of retention. Melanin was the only place of retention identified. Additional in vitro studies showed an affinity of isoxsuprine to both melanin and keratin. The K(d) values were 0.02 mmol/l and 1 mmol/l, and the B(max) values were 0.2 micromol/mg and 2 micromol/mg, respectively. A ...
van der Kolk JH, Wensing T.The urinary corticoid:creatinine (c:c) ratio was determined in ten pony mares suffering from hyperlipoproteinaemia. The mean (+/- sd) urinary c:c ratio of these ten ponies (47 +/- 31 x 10(-6)) was not significantly different from that of twelve pony mares with a pituitary pars intermedia adenoma (31 +/- 18 x 10(-6). The correlation between the urinary concentration of corticoids and plasma total lipids, and the correlation between the urinary c:c ratio and plasma total lipids in ponies with hyperlipoproteinaemia were not significant (P > 0.05; r = 0.53 and r = 0.008, respectively). Preliminary...
Schott HC, Düsterdieck KF, Eberhart SW, Woody KA, Refsal KR, Coenen M.Incomplete recovery from endurance exercise after an overnight rest period is reflected by persisting weight loss and an elevated plasma aldosterone concentration, even in successful competitors. To determine whether supplementation with high doses of electrolytes, with or without glycerol, enhances recovery, the following were measured in 6 Arabian horses before and after completion of a 60 km treadmill exercise test simulating an endurance ride and after 12, 24, 48, and 72 h of recovery: bodyweight; plasma osmolality; plasma concentrations of protein, electrolytes, aldosterone and cortisol; ...
Bailey J, Colahan P, Kubilis P, Pablo L.This study was designed to test the hypothesis that a specific beta 2 adrenergic receptor agonist, albuterol sulphate, when delivered by metered-dose inhaler (MDI) would affect the performance of Thoroughbred horses during incremental exercise testing. Six conditioned Thoroughbred horses were randomly assigned to one of 2 treatment groups: Group 1 horses received placebo and Group 2 received albuterol. Each horse received both treatments in a crossover design with a 3 week interval between trials. Horses inspired albuterol (900 micrograms) or placebo over a 5 min period followed by an incremen...
Choi MH, Kim JY, Chung BC.A highly specific method is described for measuring the testosterone:epitestosterone ratio in equine urine by gas chromatography-mass spectrometry (GC-MS) with stable isotope internal standards. The procedure was based on Serdolit Pad-1 resin extraction, enzymatic hydrolysis, and chemical derivatisation prior to instrumental analysis. The mixed derivatives, 3-trimethylsilyl-17-pentafluorophenyldimethylsilyl ether (3-TMS-17-flophemesyl) testosterone and epitestosterone, were found to have excellent analytical properties. The specificity of the derivatisation method exploits a unique feature of ...
Todi F, Mendonca M, Ryan M, Herskovits P.The origin of caffeine detections in equine serum and urine after theophylline administrations was examined. Three different preparations containing theophylline were administered to standardbred mares. Both blood and urine samples were collected. Caffeine was detected and quantified in theophylline administration samples by high performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS-MS). Further in vitro analysis showed that caffeine metabolites were not detected when caffeine, or caffeine-containing products, were added to urine. Data derived from ...
Hagedorn HW, Meiser H, Zankl H, Schulz R.The misuse of opiates in racehorses relates to their effect of increasing locomotor activity. Because methadone, a narcotic analgesic, has been suspected of use as a doping compound in the past, it was added to the list of banned drugs and should be considered in doping control. Because the literature fails to provide information on detection of methadone in blood or urine of horses, an enzyme-linked immunosorbent assay was developed to monitor this narcotic in equine body fluids. Combined with high-performance liquid chromatography, the immunoassay also served to confirm positives indicated b...
Jose-Cunilleras E, Hinchcliff KW.Pharmacologic treatment of diseases of the urinary tract of horses is limited to administration of antibiotics for treatment of urinary tract infections (UTIs), administration of drugs that alter urine pH, administration of drugs that alter bladder smooth muscle function or urethral sphincter tone, and treatment of acute renal failure. The indications, mechanisms of action, pharmacokinetic characteristics, and adverse effects of these agents in each of these groups are discussed in this article. The use of the agents is discussed within the context of the pathophysiology of the disease being t...
Freeman DA, Cymbaluk NF, Schott HC, Hinchcliff K, McDonnell SM, Kyle B.To compare health, hydration status, and management of stabled pregnant mares provided drinking water continuously or via 1 of 3 intermittent delivery systems. Methods: 22 Quarter Horse (QH) or QH-crossbred mares and 18 Belgian or Belgian-crossbred mares (study 1); 24 QH or QH-crossbred mares and 18 Belgian or Belgian-crossbred mares (study 2). Methods: Stabled horses were provided water continuously or via 1 of 3 intermittent water delivery systems in 2 study periods during a 2-year period. Body temperature, attitude, appetite, water intake, and urine output were recorded daily. Hygiene of ea...
Mirza MH, Oliver JL, Seahorn TL, Hosgood G, Moore RM.The purpose of this study was to determine whether nitric oxide (NO) is present in clinically normal horses under basal conditions and if it increases secondary to naturally acquired small intestinal strangulation obstruction. Thirty-one horses were used; 20 horses with naturally acquired small intestinal strangulation obstruction and 11 clinically normal horses with no signs of gastrointestinal tract disease. Jugular venous blood, abdominal fluid, and urine were collected for NO quantification. Plasma, abdominal fluid, and urine were stored at -70 degrees C until analyzed for NO using a chemi...
Stevenson AJ, Weber MP, Todi F, Mendonca M, Fenwick JD, Kwong E, Young L, Leavitt R, Nespolo R, Beaumier P.A study of the effects of intravenous administration of either 150 mg or 250 mg of furosemide to standardbred mares pre-treated with other drugs was undertaken to determine whether a unique pattern of drug elimination into urine and from plasma for each compound occurred. Furosemide significantly reduced the plasma concentrations of codeine compared to control 2-6 h after furosemide administration. In contrast, the plasma concentrations of theophylline, phenylbutazone, pentazocine, guaifenesin and flunixin were not markedly altered by furosemide. In the case of acepromazine, clenbuterol and fe...
Zeyner A, Kirbach H, Fürll M.Feeding a high-fat diet may have potential advantages by maintaining pH homeostasis during intense exercise; conversely, effects including elevated PCV and increased plasma concentration of total protein may indicate effects detrimental to performance by reducing perfusion of tissues and organs. Six horses were used to determine the effects of an replacement of starch (diet '0.00') with 0.33, 0.67, 1.00 and 1.33 g soybean oil/kg bwt/day (diets '033', '0.67', '1.00' and '1.33') on venous and renal acid-base status and fractional clearance (FC) of electrolytes in a postprandial state but prior t...
Teale P, Houghton E.A screening procedure for anabolic steroid residues in horse urine has been developed based upon solid-phase extraction and gas chromatographic/mass spectrometric analysis in the selected ion mode. For moderate sample throughput the method provides a viable alternative to radioimmunoassay screening and has advantages over the latter technique due to its flexibility, specificity and ability to detect a number of steroids in a single analysis. Full automation of the gas chromatographic/mass spectrometric analysis is an additional feature of the methodology.
Uberti B, Pressler BM, Alkabes SB, Chang CY, Moore GE, Lescun TB, Sojka JE.To investigate the effects of heparin administration on urine protein excretion during the developmental stages of experimentally induced laminitis in horses. Methods: 13 horses. Procedures-Horses received unfractionated heparin (80 U/kg, SC, q 8 h; n=7) or no treatment (control group; 6) beginning 3 days prior to induction of laminitis. All horses were given 3 oligofructose loading doses (1 g/kg each) at 24-hour intervals and a laminitis induction dose (10 g of oligofructose/kg) 24 hours following the final loading dose (designated as 0 hours) via nasogastric tube. Serum glucose and insulin c...
Knych HK, McKemie DS, Seminoff K, Hartmann P, Hovda L, Benson D.Ethylphenidate is a psychostimulant and analog of the commonly prescribed compound, methylphenidate (Ritalin®). There are a limited number of studies describing the disposition and pharmacologic/toxicological effects of ethylphenidate in any species. The abuse potential in equine athletes along with the limited data available regarding administration in horses necessitates further study. The objectives of the current study were to describe drug concentrations, develop an analytical method that could be used to regulate its use, and describe the pharmacodynamic effects of ethylphenidate in hor...
Neto LM, Andraus MH, Salvadori MC.A method is described for the qualitative and quantitative determination of phenylbutazone and oxyphenbutazone in horse urine and plasma samples viewing antidoping control. A horse was administered intravenously with 3 g of phenylbutazone. For the qualitative determination, a screening by HPLC was performed after acidic extraction of the urine samples and the confirmation process was realized by GC-MS. Using the proposed method it was possible to detect phenylbutazone and oxyphenbutazone in urine for up to 48 and 120 h, respectively. For the quantitation of these drugs the plasma was deprotein...
Fidani M, Pompa G, Mungiguerra F, Casati A, Fracchiolla ML, Arioli F.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...
Greene EW, Woods WE, Tobin T.Caffeine (4 mg/kg) was given by rapid IV injection to 4 horses. Plasma concentrations of the drug peaked at 10 micrograms/ml and decreased rapidly at first, and then more slowly, with an apparent beta-phase half-life of 18.2 hours. Urinary concentrations of caffeine were remarkably consistent at about 3 times plasma values of the drug. Caffeine was detectable in both plasma and urine of the horses for up to 9 days after dosing. After oral administration, caffeine was absorbed poorly with an apparent bioavailability of 39%. Although blood concentrations of caffeine peaked rapidly after oral adm...
Hagedorn HW, Zuck S, Schulz R.The azaphenothiazine neuroleptic prothipendyl (Dominal) is suspected to be administered illegally at low doses to race-horses to improve their performance. Since for this species pharmacokinetic data of the drug are missing we studied its elimination from blood and urine in a standard-bred mare. At a low (subtherapeutic) dose (i.v., 0.24 mg/kg) the horse is described to be less excited while locomotor activity and attention remain unaffected. In contrast, sedation and ataxia are brought about at 1 mg/kg (therapeutic dose). Identification of prothipendyl given i.v. at subtherapeutic doses was a...
Brown MP, Stover SM, Kelly RH, Farver TB.Ten healthy adult mares were given a single intramuscular dose (2.2 mg/kg) of gentamicin sulfate. Over a 48-h period, gentamicin concentrations were measured serially in the serum of all ten mares and in synovial fluid, peritoneal fluid, and urine of six of the mares. The mean peak serum gentamicin concentration was 5.73 micrograms/ml at 1 h. Gentamicin was detected in synovial fluid and peritoneal fluid, with mean peak gentamicin concentrations of 2.41 micrograms/ml and 3.92 micrograms/ml, respectively, observed at 2 h. These concentrations declined in parallel with serum concentrations and w...
Tang FP, Leung GN, Wan TS.A capillary electrophoresis-mass spectrometry (CE-MS) method for the analysis of quaternary ammonium drugs in equine urine was developed. Quaternary ammonium drugs were first extracted from equine urine by ion-pair extraction and then analysed by CE-MS in the positive electrospray ionization (ESI) mode. Within 12 min, eight quaternary ammonium drugs, each at 1 ng/mL in horse urine, could be detected. The confirmation of these drugs in urine samples was achieved by capillary electrophoresis tandem mass spectrometry (CE-MS/MS). A direct comparison of this method was made with existing liquid chr...
Mizobe F, Mori M, Nagata SI, Yamashita S, Okada J, Kusano K.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 us...
Saikku A, Koskinen E, Sandholm M.IgG levels and tryptic inhibition were investigated in sequentially collected mare's colostrum and milk, foal serum and urine. The colostral trypsin-inhibitor was "transfused" to the newborn foal by the colostral intestinal route in parallel with IgG. However, the trypsin-inhibitor as a small molecular weight inhibitor became excreted into urine peaking at about 20 hours. The physiological proteinuria in foals during the first 2 days is mostly due to immunoglobulin fragments and colostral-derived trypsin-inhibitor. Analysis of urine for IgG light chains or trypsin inhibitor will therefore reve...
Harding C, Viljanto M, Cutler C, Habershon-Butcher J, Biddle S, Scarth J.Oxandrolone is an anabolic-androgenic steroid with favourable anabolic to androgenic ratio, making it an effective anabolic agent with less androgenic side effects. Although its metabolism has been studied in humans, its phase I and II metabolism has not been previously reported in the horse. The purpose of this study was to investigate the in vitro metabolism of oxandrolone (using both equine liver microsomes and S9) and in vivo metabolism following oral administration (three daily doses of 50 mg of oxandrolone to a single Thoroughbred horse), using both gas and liquid chromatography-mass s...
Houston T, Chay S, Woods WE, Combs G, Kamerling S, Blake JW, Edmundson AG, Vessiney R, Tobin T.A survey of plasma and urinary concentrations of phenylbutazone and its metabolites in thoroughbred horses racing in Kentucky was carried out. Post-race blood samples from more than 200 horses running at Latonia Racetrack and Keeneland in the Spring of 1983 were analysed. The modal plasma concentration of phenylbutazone was between 1 and 2 micrograms/ml, the mean concentration was 3.5 micrograms/ml and the range was up to 15 micrograms/ml. Oxyphenbutazone had a modal plasma concentration between 1 and 2 micrograms/ml, a mean concentration of 2.07 micrograms/ml and a range of up to 13 microgram...
Cymbaluk NF, Schryver HF, Hintz HF, Smith DF, Lowe JE.The effects of molybdenum ingestion on copper metabolism were investigated in bile duct-cannulated ponies fed diets containing 1.01, 27.4 and 107.3 ppm molybdenum. Copper metabolism was assessed by examination of 64Cu and stable copper excretion in feces, bile and urine. The addition of molybdenum to the diet decreased copper absorption and retention as a consequence of increased excretion of dietary copper in feces and increased excretion of absorbed copper in bile. Only 5% or less of the absorbed copper was excreted in urine regardless of dietary treatment. Biliary molybdenum excretion was o...
Tsitsimpikou C, Spyridaki MH, Georgoulakis I, Kouretas D, Konstantinidou M, Georgakopoulos CG.Flurbiprofen and its main acidic metabolites were detected in equine urine after a single-dose administration of 500 mg flurbiprofen to two 2.5-3.5-years-old mares, in order to be used in equine doping control routine analysis. The urine levels of the parent drug were determined using GC/MS. Five acidic metabolites were found in the urine. The structure of the proposed metabolites was confirmed by HRMS accurate mass measurements. The highest flurbiprofen concentration was 204 mug ml(-1) at 1-3 h post administration. Flurbiprofen could be detected for 24-37 h in urine using the standard screeni...
Gilliam LL, Ownby CL, McFarlane D, Canida A, Holbrook TC, Payton ME, Krehbiel CR.Rattlesnake bites in horses are not uncommon and the clinical outcomes are widely variable. Treatment of horses with anti-venom is often cost prohibitive and could have negative consequences; therefore, the development of a quantitative test to determine if anti-venom therapy is indicated would be valuable. The objective of this study was to develop an ELISA to detect rattlesnake venom in biological samples from clinically bitten horses. Nineteen horses were enrolled in the study. Urine was available from 19 horses and bite site samples were available from 9 horses. A double sandwich fluoresce...
Snow DH, Munro CD, Nimmo MA.The effect of 11 weekly injections of nandrolone phenylpropionate (400 mg) was investigated by a crossover trial (2 training periods) in 6 Thoroughbred geldings undergoing training. A decrease in body weight and flank measurement occurred only during the first training period and was not modified by the anabolic steroid. Urinary nitrogen excretion was lower in the anabolic treated animals only in the first training period. Neither training nor training plus nandrolone phenylpropionate administration caused any marked alteration in blood biochemistry or haematology. A significant decrease in pl...
Menendez MI, Phelps MA, Bertone AL.To the date, no reports exist of the pharmacokinetics (PK) of betamethasone (BTM) sodium phosphate and betamethasone acetate administered intra-articular (IA) into multiple joints in exercising horses. The purpose of the study was to determine the PK of BTM and HYD concentrations in plasma and urine after IA administration of a total of 30 mg BTM. Eight 4 years old Thoroughbred mares were exercised on a treadmill and BTM was administered IA. Plasma and urine BTM and HYD were determined via high performance liquid chromatography spectrometry for 6 weeks. Concentration-time profiles of BTM and H...
Soma LR, Gallis DE, Davis WL, Cochran TA, Woodward CB.Phenylbutazone (PBZ) was administered (8.8 mg/kg of body weight) every 24 hours for 5 consecutive days, orally for the first 4 days and IV on day 5. The half-life (t 1/2) after this daily administration was 6.2 hours and the volume of distribution was 0.152 +/- 0.014 L/kg; the bioavailability after oral administration was 91.8 +/- 2.5%. The plasma concentration of PBZ at experimental hour (EH) 24 (24 hours after the 1st oral dose) was 1.7 +/- 0.39 micrograms/ml and increased to 4.2 +/- 0.29 micrograms/ml at EH 48 (24 hours after the 2nd oral dose). Values at EH 72, 96, and 120 (24 hours after ...
Zwain I, Gaillard JL, Dintinger T, Silberzahn P.A single i.m. injection of testosterone (750 mg of testosterone bexahydrobenzoate) or i.v. injection of human chorionic gonadotrophin (hCG) (10,000 IU) was given to geldings and stallions. Levels of unconjugated and conjugated (after solvolysis) androgens and estrogens were measured in blood and urine samples taken daily from the day of injection (D0) to the tenth day post-injection (D10). In the stallion, both treatments resulted in a sharp increase of plasma estrogens, which peaked one day before the androgen levels. Our results confirmed the testicular localization of a potent aromatase, wh...
Yi R, Sandhu J, Zhao S, Lam G, Loganathan D, Morrissey B.Efaproxiral (RSR 13) is an experimental synthetic allosteric modifier of haemoglobin (Hb) that acts by increasing the release of oxygen from Hb to the surrounding tissues. It has been shown to increase maximum oxygen uptake (VO(2max)) in a canine skeletal muscle model. The ability to increase maximal muscle oxygen uptake makes efaproxiral a potential performance-enhancing agent and is therefore prohibited by the World Anti-Doping Agency. In this study, a method for the detection and elimination of efaproxiral in equine plasma and urine after a 2.5 g intravenous administration of efaproxiral ...
Gadaj A, Ventura E, Healy J, Botrè F, Sterk SS, Buckley T, Mooney MH.Selective androgen receptor modulators (SARMs) represent non-steroidal agents commonly abused in human and animal (i.e. equine, canine) sports, with potential for further misuse as growth promoting agents in livestock-based farming. As a direct response to the real and possible implications of illicit application in both sport as well as food production systems, this study incorporated enzymatic hydrolysis (β-glucuronidase/arylsulfatase) into a previously established protocol while maintaining the minimal volume (200 µL) of urine sample required to detect SARMs encompassing various pharmacop...
Bhavnani BR, Woolever CA.[3H]Equilin [3H-labeled 3-hydroxy-1,3,5(10), 7-estratetraen-17-one] was administered iv to a pregnant mare in the 10th month of gestation. Maternal urine was collected for 3 days, and blood samples were taken 35 min and 3, 6, 12, and 24 h after the injection. The half-life of the disappearance of radioactivity from the blood was approximately 2.5 h. Over 90% of the administered dose was excreted in the first 24 h. The urine was extracted, hydrolyzed, and fractionated. The bulk of the radioactive material (75%) was present in the phenolic sulfate fraction from which radiochemically pure equilin...
Houghton E, Dumasia MC, Wellby JK.Negative ion chemical ionization mass spectra of some corticosteroids have been obtained by direct syringe introduction on to the Finnigan moving belt high-performance liquid chromatography-mass spectrometer interface. Proprietary preparations based upon dexamethasone, betamethasone and prednisolone were administered to horses at therapeutic dose level. Urine samples were extracted, the extracts purified by Sephadex LH-20 chromatography and the presence of the parent steroids in the eluates was confirmed by combined high-performance liquid chromatography negative ion chemical ionization mass s...
McGorum BC, Anderson RA.To test the hypothesis that equine grass sickness may be associated with the ingestion of cyanogenic glycosides from white clover (Trifolium repens), the concentrations of whole blood cyanide, and plasma and urinary thiocyanate, the main metabolite of cyanide, were measured in 12 horses with acute grass sickness and 10 horses with subacute grass sickness, and in 43 control horses, of which 21 were co-grazing with cases of acute grass sickness, 12 grazed pastures where grass sickness had not been reported, and 10 were stabled horses. The healthy horses which grazed with cases of acute grass sic...
Bailey J, Colahan P, Kubilis P, Pablo L.This study was designed to test the hypothesis that a specific beta 2 adrenergic receptor agonist, albuterol sulphate, when delivered by metered-dose inhaler (MDI) would affect the performance of Thoroughbred horses during incremental exercise testing. Six conditioned Thoroughbred horses were randomly assigned to one of 2 treatment groups: Group 1 horses received placebo and Group 2 received albuterol. Each horse received both treatments in a crossover design with a 3 week interval between trials. Horses inspired albuterol (900 micrograms) or placebo over a 5 min period followed by an incremen...
Johnson E, van Heemst J, Benavides J, Gray B.Etizolam is a benzodiazepine analogue that is approved for use in Japan, Italy and India but has recently appeared as a nonapproved product on the illicit drug market in Europe and North America. Etizolam was identified in a crystalline material seized at a Kentucky racetrack, raising concerns that this drug may have been used in racing. The aim of this study was to characterize the metabolism and excretion of etizolam in horses to generate information on its disposition and to incorporate the correct urinary and serum target analytes into anti-doping screening procedures. Etizolam was adminis...
McKeever KH, Scali R, Geiser S, Kearns CF.The purpose of the present study was to determine whether the training-induced hypervolaemic response seen in the horse is associated with aldosterone-mediated renal mechanisms affecting sodium conservation during the first days of training. Five healthy, Standardbred mares (weight 450-500 kg, age 4-8 years) that were unfit, but accustomed to running on the treadmill, were used to test the hypothesis that repeated submaximal exercise would alter plasma aldosterone (ALDO) concentration and renal excretion of electrolytes in horses within the first 3 days of training. The experiment consisted of...
