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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.
Determination of succinonitrile in horse urine by gas chromatography-nitrogen-phosphorus detector and gas chromatography-mass spectrometry. A chromatographic method was developed to detect and confirm the presence of succinonitrile (SDN) in horse urine samples, for antidoping control. The urine samples (5 ml) were extracted with diethyl ether and screened by gas chromatography-nitrogen-phosphorus detector and the confirmation of the drug's presence was accomplished by using gas chromatography-mass selective detection. The recovery of extraction was 78 and 81% for 1.0 and 2.0 micrograms ml-1 (relative standard deviation, < 10%), respectively. Urine samples collected after the administration of Energisan were positive for SDN (1-30 ...
Preliminary study of the metabolism of 17 alpha-methyltestosterone in horses utilizing gas chromatography-mass spectrometric techniques. Little is known about the metabolism of 17 alpha-alkyl anabolic steroids in horses. In this study, the metabolism of 17 alpha-methyltestosterone is investigated by oral administration of a (1 + 1) mixture of the steroid and its deuteriated analogue. Both compounds were synthesized from dehydroisoandrosterone (DHA), using a Grignard reaction followed by an Oppenauer oxidation. Post-administration urine extracts were analysed by gas chromatography--mass spectrometry (GC-MS) using both electron impact (IE) and chemical ionization (CI). Interpretation of the data was facilitated by observation of ...
Generic immunoassay of corticosteroids with minimum pre-treatment of urine samples. A generic, rapid and sensitive enzyme linked immunosorbent assay (ELISA) test has been developed which allows large-scale simultaneous testing of synthetic corticosteroids viz., flumethasone, dexamethasone and betamethasone. This assay can be directly applied to diluted urine samples (1 + 9) without hydrolysis of glucuronide or sulfate conjugates or any other treatment of samples. The polyclonal antibody was obtained by immunizing sheep with a flumethasone derivative linked to human serum albumin. This polyclonal antibody displayed high-reactivity with several synthetic corticosteroids whilst ...
Screening of non-steroidal anti-inflammatory drugs, barbiturates and methyl xanthines in equine urine by gas chromatography-mass spectrometry. A gas chromatographic-mass spectrometric (GC-MS) screening procedure for 23 acidic drugs in equine urine is described. With the GC-MS method fifteen anti-inflammatory drugs, five barbiturates and three methyl xanthines can be detected with good sensitivity and selectivity. The method consists of alkaline hydrolysis, extraction with organic solvent using salting-out, clean-up extraction, methylation and screening with GC-MS in selected-ion monitoring mode. The limit of detection is 10 micrograms 1(-1) or lower, for most drugs.
Endogenic nortestosterone in cattle? When residues of nortestosterone (NT) were found in the urine of cattle, racehorses or bodybuilders, exogenic administration was thought to be proven. In previous literature, no records were found of the endogenic presence of this molecule. In the horse-racing world, Houghton and Courthot found that NT is normally present in the urine of the stallion. Belgian and Dutch researchers found that NT is also present in the urine and edible parts of the intact boar. Vandenbroeck et al. (1991) suggested the endogenous presence of NT (in the beta form) in the pregnant cow. Meyer (1992) reported the pre...
[Renal tubular acidosis (type 2) in a Friesian mare]. A 2-year-old Friesian mare was examined because she had been sluggish for a few weeks. Blood biochemistry showed that concentrations of creatinine (136 mumol.L-1) and urea nitrogen (6.1 mmol.L-1) in plasma were within their normal limits. Severe metabolic acidosis was apparent; the venous pH was 7.154 and the bicarbonate concentration was 13 mmol.L-1. The base excess amounted to -15.3 mmol.L-1 and the venous pCO2 measured 5.1 kPa. Plasma sodium was normal (135 mmol.L-1) and plasma chloride (113 mmol.L-1) was slightly elevated, but there was a hypokalaemia (2.9 mmol.L-1). The urine had a specif...
Disposition of human drug preparations in the horse. III. Orally administered alclofenac. Concentrations of the non-steroidal anti-inflammatory drug (NSAID) alclofenac were determined by a sensitive high performance liquid chromatographic procedure in plasma and urine of horses following oral administration of a dose of 3 g. In plasma, alclofenac was present in detectable concentrations for 72 h. The plasma disposition in individual horses was best described by a bi-compartmental model with two successive rate constants ka1 = 0.05 +/- 0.06 h-1 and ka2 = 0.06 +/- 0.01 h-1. Alclofenac half-lives t1/2 alpha and t1/2 beta were 1.0 +/- 0.8 h and 6.9 +/- 1.5 h, respectively. Maximal conc...
Monitoring furosemide in racehorses participating in an EIPH program. Analytical procedures were developed to monitor furosemide concentrations in post-race serum and urine samples obtained from horses participating in an exercise-induced pulmonary haemorrhage (EIPH) program. High performance liquid chromatography with ultraviolet light detection proved a reliable, sensitive method for measuring urinary furosemide concentrations up to 12 h after administration of either 150 or 250 mg of the drug to race horses. However, this method was unreliable for determination of serum furosemide concentration. High performance liquid chromatography with fluorescence detecti...
Immunoassay detection of drugs in racing horses: detection of ethacrynic acid and bumetanide in equine urine by ELISA. We have raised antibodies and developed one-step enzyme-linked immunosorbent assays (ELISA) for the diuretics ethacrynic acid and bumetanide as part of a panel of pre- and post-race tests for high potency drugs in racing horses. These ELISA tests are rapid (completed within one hour), sensitive, and can be read by eye. The ELISA detects ethacrynic acid at a drug concentration for half-maximal inhibition (I-50) of about 2.5 ng/mL for the parent drug. After dosing horses intravenously with 5 mg ethacrynic acid per horse, the parent drug or its metabolites are detectable in urine for at least 8 h...
Serological and microbiological findings on 3 farms with equine leptospiral abortions. Blood and urine samples from horses on 3 central Kentucky horse farms with prior histories of leptospiral abortions were analysed. Blood samples were obtained from all available horses on each farm and tested for antibodies to 6 leptospira serovars. Urine samples were collected from non-gravid mares with serum antibody titres > or = 1:800 and examined for leptospires by dark-field microscopy, fluorescent antibody testing and culture. Adult horses had the greatest serological evidence of exposure to leptospira, followed by yearlings, then foals. Of horses with anti-leptospiral antibodies, 76...
Identification and verification of the anabolic steroid boldenone in equine blood and urine by HPLC/ELISA. An enzyme linked immunosorbent assay (ELISA) was developed to detect the anabolic steroid boldenone in equine blood and urine. The polyclonal antiserum was raised in rabbits, employing boldenone-17-hemisuccinate-bovine serum albumin as antigen. Boldenone-17-hemisuccinate-horseradish peroxidase served as enzyme conjugate. Sensitivity of the assay was 26.0 +/- 3.0 pg/well. Among the endogenous steroids tested only progesterone and testosterone exhibited moderate cross-reactivities, 3.4 and 2.5%, respectively. These cross-reactivities are of no importance for the boldenone assay. For the reductio...
Investigation of the metabolism of azaperone in the horse. Urine samples collected from a horse after intramuscular administration of 40 mg of azaperone were extracted at pH 10 before and after acid hydrolysis. The extracts were concentrated and analysed by LC-MS-MS. Two N-dealkylated metabolites, N-despyridinylazaperol and N-despyridinylazaperone, and a low concentration of azaperone were detected in the unhydrolysed urine. Six metabolites; hydroxyazaperol, two hydroxyazaperones, azaperol, N-despyridinylazaperol and N-despyridinylazaperone were detected in the hydrolysed urine extracts. Using XAD-2 resin extraction, three glucuronide conjugated azape...
Measurements of urethral pressure profiles in the male horse. Urinary tract pressure profiles were evaluated in 6 clinically normal geldings over 3 consecutive days. This was performed by introducing a 1.3 m-long cuffed catheter into the urinary tract, under xylazine sedation (0.8 mg/kg, iv). The method was reproducible. The mean (+/- sd) intra-vesicular pressure (IVP) and maximal urethral closure pressures (MUCP) were 10.3 (+/- 1.7) and 129.8 (+/- 19.6) cmH2O, respectively, and the ratio between MUCP and IVP was 13.2 (+/- 2.5). A gelding with urinary incontinence showed a significantly lower MUCP (73.4 cmH2O), and an MUCP to IVP ratio of 8.0. It was con...
A liquid chromatographic method for the determination of fenoprofen in equine plasma and urine. A high performance liquid chromatographic method to measure plasma and urine fenoprofen levels in equine biofluids is described. Liquid-liquid extraction with diethylether was used to isolate the drug from plasma and urine. The accuracy and reproducibility of the method were within acceptable limits over the concentration range 0-10 micrograms/mL and 0-20 micrograms/mL respectively from plasma and urine. Detection limits were 0.05 microgram/mL (2 mL plasma) and 0.2 microgram/mL (0.5 mL urine). This procedure was applied to ascertain the pharmacokinetics of a 3 g dose of fenoprofen calcium in a...
Determination of alclofenac in equine plasma and urine by high-performance liquid chromatography. A high-performance liquid chromatographic method to measure plasma and urinary alclofenac levels in equine biofluids is described. Isolation of the drug from plasma is achieved using liquid-liquid extraction with diethyl ether. Reversed-phase C18 solid phase extraction is used for the extraction of free and conjugated alclofenac from urine. The reproducibility and accuracy of the method were well within acceptable limits over the concentration ranges 0-10 and 0-20 micrograms/ml, respectively, for plasma and urine. Starting with 2 ml of plasma, a concentration of 0.1 microgram/ml could easily b...
Comparison of immunoaffinity chromatography combined with gas chromatography-negative ion chemical ionisation mass spectrometry and radioimmunoassay for screening dexamethasone in equine urine. A comparison of the sensitive analytical methods of radioimmunoassay (RIA) and immunoaffinity chromatography (IAC) combined with gas chromatography-negative ion chemical ionisation mass spectrometry for the specific and reliable screening of dexamethasone in equine post-race urine is presented. Results from analyses of samples collected from a mare during 144 hours post administration of 26 mg of dexamethasone sodium phosphate are described.
The disposition of suxibuzone in the horse. A high performance liquid chromatographic method is described to determine the anti-inflammatory drug suxibuzone (SXB) and its major metabolites phenylbutazone (PBZ) and oxyphenbutazone (OPBZ) in equine plasma and urine. When suxibuzone (6 mg/kg) was administered intravenously (i.v.) or orally (p.o.) no parent drug was detected in plasma or in urine. The disposition of the metabolite PBZ (i.v.) could be described by a 2 compartment model with a beta half-life varying from 7.40 to 8.35 h. Due to severe side effects the use of i.v. suxibuzone should not be encouraged in the horse. PBZ and OPBZ w...
Hematuria and leptospiruria in a foal. A Thoroughbred foal that was born after 305 days of gestation was referred 6 hours after birth. On initial examination, the foal was alert, but weak and unable to stand. Heart rate was 150 beats/min, respiratory rate was 48 breaths/min, and rectal temperature was 33 C. Leukocytosis was detected (26,000 WBC/microliters). Dysuria and passage of red urine was observed approximately 30 hours after admission. Urine collected by free catch had a 3+ reaction for blood on dipstick analysis. Examination of Warthin Starry-stained sections of allantochorion revealed organisms morphologically characterist...
Potassium concentrations in muscle, plasma and erythrocytes and urinary fractional excretion in normal horses and those with chronic intermittent exercise-associated rhabdomyolysis. Potassium concentrations were measured in semimembranosus muscle, plasma and erythrocytes, and the urinary fractional excretion determined in normal horses and those that had chronic intermittent exercise-associated rhabdomyolysis. Muscle from the rhabdomyolysis horses was also evaluated microscopically. The horses with rhabdomyolysis had a lower muscle potassium concentration on a dry weight basis. Although the wet weight potassium content was also lower, the difference was not significant. Urinary fractional excretion of potassium (and also sodium and chloride) did not differ significantly b...
Pharmacokinetics of single intravenous and single and multiple dose oral administration of rifampin in mares. The disposition of rifampin in six healthy mares after single intravenous (i.v.) and oral (p.o.) doses and after seven oral doses of 10 mg/kg administered twice a day was investigated using a high performance liquid chromatographic (HPLC) method. Pharmacokinetic variables for rifampin determined using the HPLC method were comparable to variables reported from earlier studies utilizing a microbiological assay. Desascetylrifampin, a major metabolite of the parent compound, could not be detected in the serum but was detected at low concentrations in urine. Mean trough concentrations of rifampin i...
A liquid chromatographic procedure for the analysis of yohimbine in equine serum and urine. A standardbred mare was dosed with 40 mg yohimbine intravenously. Serum and urine samples were collected and analyzed for yohimbine using solvent extraction and reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Maximum yohimbine concentrations of 45 and 18 ng/mL were observed in serum and urine samples, respectively. Elimination was rapid, with half-lives of approximately 20 and 53 min observed for serum and urine, respectively. The presence of yohimbine in these samples was confirmed by liquid chromatography/mass spectroscopy (LC/MS/MS).
Immunoaffinity chromatography combined with gas chromatography-negative ion chemical ionisation mass spectrometry for the confirmation of flumethasone abuse in the equine. Immunoaffinity chromatography using a synthesised immunosorbent was used to extract tritiated dexamethasone (with dexamethasone carrier) from equine urine at a recovery of 81.7 +/- 8.4% (mean +/- S.D.). A method utilising this procedure coupled to cool on-column injection gas chromatography-negative ion chemical ionisation mass spectrometry is also described for the confirmation of low levels of flumethasone in equine urine samples.
Leptospiral abortion and leptospiruria in horses from the same farm. Leptospirosis was documented as the cause of abortion in a 5-year-old mare. Leptospires were detected in tissue specimens from fetal kidneys and from placenta by histologic evaluation of silver-stained sections. Antibodies against Leptospira interrogans serovar pomona were detected in fetal serum at a titer of 1,600 by use of a microscopic agglutination test. The mare had serum titers of 6,400; 0; 400; 800; 3,200; and 6,400 to L interrogans serovars bratislava, canicola, grippotyphosa, hardjo, icterohaemorrhagiae, and pomona, respectively. A serologic survey identified titers of at least 6,400...
Pharmacokinetics and concentrations of ceftiofur sodium in body fluids and endometrium after repeated intramuscular injections in mares. Each of 5 healthy mares was given 5 consecutive IM injections of ceftiofur sodium (2 mg/kg of body weight; 50 mg/ml) at 12-hour intervals. Ceftiofur concentrations were measured serially in serum, synovial fluid, peritoneal fluid, and urine, and were measured in CSF and endometrial tissue after the fifth dose. Mean elimination rate constant was 0.354 +/- 0.101 h-1 and elimination half-life was 2.49 +/- 0.49 hour. Mean serum ceftiofur concentrations peaked approximately 1 hour after each injection. The highest mean ceftiofur concentration was 5.09 micrograms/ml at 1 hour after the fifth dose fo...
Nephrogenic diabetes insipidus in sibling colts. Nephrogenic diabetes insipidus was diagnosed in two full sibling Thoroughbred colts. Each colt had a history of excessive urination. Extreme polydipsia (greater than 80 L per day) was documented in both colts. Inability to concentrate urine in response to water deprivation, infusion of hypertonic saline, or exogenous vasopressin administration indicated insensitivity of the collecting duct epithelial cells to vasopressin. A diagnosis of nephrogenic diabetes insipidus was further supported by a normal increase in plasma vasopressin concentration after water deprivation in the one colt in which ...
Alterations of fluid and electrolyte balance in thoroughbred racehorses following strenuous exercise during training. Alterations of acid-base status, and fluid and electrolyte balance subsequent to exercise in Thoroughbred racehorses in North America have not been well-characterized. Des-cribed here are the results of an observational study conducted to characterize changes in fluid and electrolytes following strenuous exercise of 16 Thoroughbreds under routine training conditions. Changes following strenuous exercise were determined for the following variables: serum concentrations of sodium (Na), potassium (K), chloride (Cl) and protein; pH of blood; osmolality of plasma and urine; body weight; and, fracti...
Kinetic analysis of D-xylose distribution after intravenous administration to mares. Multicompartmental analysis was applied to study the kinetics of D-xylose distribution after IV administration to healthy mares deprived of food for 12 and 96 hours. Urinary excretion of D-xylose was measured over a 15-hour period after administration. The plasma D-xylose concentrations in this study were in the range found after oral tolerance testing. The disposition of D-xylose was described by a two-compartment model with linear kinetic characteristics. Total volume of distribution decreased significantly (P < 0.025) from 0.270 L/kg of body weight after the 12-hour period of food depriv...
Identification of a benzhydrolic metabolite of ketoprofen in horses by gas chromatography-mass spectrometry and high-performance liquid chromatography. A benzhydrolic metabolite of ketoprofen, formed by reduction of the keto group of the drug, has been identified by gas chromatography-mass spectrometry in equine plasma and urine. After partial synthesis, its structure has been confirmed by UV, IR and 1H NMR spectroscopy. The kinetics of ketoprofen and this metabolite have been monitored in plasma by high-performance liquid chromatography. The two products were quantified in plasma up to 4 and 3 h, respectively, and were detected in urine up to 72 and 24 h, respectively, after a single intravenous administration to horses at the dose of 2.2 mg...
Renal dysfunction associated with infection of Leptospira interrogans in a horse. Renal failure associated with infection of Leptospira interrogans was detected in a horse. Fever, leukocytosis, pyuria, isosthenuria, and azotemia were suggestive of an inflammatory urinary tract disease. Despite persistent pyuria, no bacteria were found during routine microscopic examinations or bacteriologic culturing of urine. A fluorescent antibody examination of the urine was positive for L interrogans. Serologic testing during a 6-month period, supported an acute infection with L interrogans serovar pomona. Treatment with intravenously administered fluids and antimicrobials resulted in c...
Characterization of metabolites of xylazine produced in vivo and in vitro by LC/MS/MS and by GC/MS. The metabolic fate of xylazine, 2-(2,6-dimethylphenylamino)-5,6-dihydro-4H-1,3-thiazine, in horses is described. The major metabolites identified in the hydrolyzed horse urine were 2-(4'-hydroxy-2',6'-dimethylphenylamino)-5,6-dihydro-4H-1,3-thiazi ne, 2-(3'-hydroxy-2',6'-dimethylphenylamino)-5,6-dihydro-4H-1,3-thiazi ne, N-(2,6-dimethylphenyl)thiourea, and 2-(2',6'-dimethylphenylamino)-4-oxo-5,6-dihydro-1,3-thiazine. These metabolites were also produced by incubating xylazine with rat liver microsomes. The major metabolite produced in vitro by rat liver preparations was found to be the ring op...
