Detection, quantification, and pharmacokinetics of furosemide and its effects on urinary specific gravity following IV administration to horses

Furosemide is a potent loop diuretic used for the prevention of exercise-induced pulmonary hemorrhage in horses. This drug may interfere with the detection of other substances by reducing urinary concentrations, so its use is strictly regulated. The regulation of furosemide in many racing jurisdictions is based on paired limits of urinary SG (<1.010) and serum furosemide concentrations (>100 ng/ml). To validate this regulatory mechanism, a liquid chromatography/mass spectrometry/mass spectrometry method employing a solid-phase extraction procedure and furosemide-d5 as an internal standard was developed. The method was used to determine the pharmacokinetic parameters of furosemide in equine serum samples and its effects on urinary SG after IV administration (250 mg) to 10 horses. Pharmacokinetic analysis showed that serum concentrations of furosemide were well described by a two-compartmental open model. Based on results in this study, it is very unlikely for horses to have serum furosemide concentrations greater than 100 ng/ml or urine SG less than 1.010 at 4 hours after administration (250 mg IV). However, it should be remembered that urine SG is a highly variable measurement in horses, and even without furosemide administration, some horses might naturally have urine SG values less than 1.010.     

Lidocaine in the horse: its pharmacological effects and their relationship to analytical findings

Lidocaine is a local anaesthetic agent that is widely used in equine medicine. It is also an Association of Racing Commissioners International (ARCI) Class 2 foreign substance that may cause regulators to impose substantial penalties if residues are identified in post race urine samples. Therefore, an analytical/pharmacological database was developed for this drug. Using our abaxial sesamoid local anaesthetic model, the highest no-effect dose (HNED) for the local anaesthetic effect of lidocaine was determined to be 4 mg. Using enzyme-linked immunosorbent assay (ELISA) screening, administration of the HNED of lidocaine to eight horses yielded peak serum and urine concentrations of apparent lidocaine of 0.84 ng/mL at 30 min and 72.8 ng/mL at 60 min after injection, respectively. These concentrations of apparent lidocaine are readily detectable by routine ELISA screening tests (LIDOCAINE ELISA, Neogen, Lexington, KY). ELISA screening does not specifically identify lidocaine or its metabolites, which include 3-hydroxylidocaine, dimethylaniline, 4-hydroxydimethylaniline, monoethylglycinexylidine, 3-hydroxymonoethylglycinexylidine, and glycinexylidine. As 3-hydroxylidocaine is the major metabolite recovered from equine urine, it was synthesized, purified and characterized, and a quantitative mass spectrometric method was developed for 3-hydroxylidocaine as recovered from horse urine. Following subcutaneous (s.c.) injection of the HNED of lidocaine, the concentration of 3-hydroxylidocaine recovered from urine reached a peak of about 315 ng/mL at 1 h after administration. The mean pH of the 1 h post dosing urine samples was 7.7, and there was no apparent effect of pH on the amount of 3-hydroxylidocaine recovered. Within the context of these experiments, the data suggests that recovery of less than 315 ng/mL of 3-hydroxylidocaine from a post race urine sample is unlikely to be associated with a recent local anaesthetic effect of lidocaine. Therefore these data may be of assistance to industry professionals in evaluating the significance of small concentrations of lidocaine or its metabolites in postrace urine samples. It should be noted that the quantitative data are based on analytical methods developed specifically for this study, and that methods used by other laboratories may yield different recoveries of urine 3-hydroxylidocaine.     

Feedlot performance of Brahman x Angus versus Angus steers during cold weather

Ten Angus and 10 Brahman x Angus F1 steers were used in a 184-d trial to compare feedlot performance during cold weather (-9 to 26 degrees C). Both groups of steers were exposed to the same environment for the same amount of time. All steers were fed for the same number of days regardless of frame score to avoid frame score x environment interactions. Brahman x Angus steers were 30.7 kg heavier (P less than .05) than Angus steers at the start of the trial. Differences in age (Brahman x Angus 40 d younger) for the two breed groups did not affect final live weight or carcass weight. Brahman x Angus steers consumed .2% less feed (P less than .05) as a percentage of BW than Angus steers; however, there was no difference in overall feed efficiency. Angus steers had a higher yield grade, more fat at the 12th rib (P less than .05), and graded 90% Choice; only 10% of the Brahman x Angus were graded Choice. Brahman x Angus steers were taller at the hip (P less than .05) and longer from first rib to aitch bone (P less than .05) and from thoracic vertebrae (T12/T13) to point of hock (P less than .05). Hide thickness determined at the neck, belly, and rump was found to be similar (7.7 mm) between the two groups. Sample hair weight and diameter did not differ between groups. Fiber, fat, protein, and DM digestibility coefficients were similar between groups but Brahman x Angus feces had a higher DM content.(ABSTRACT TRUNCATED AT 250 WORDS)     

Apparent ELISA detection times for albuterol after administration with the torpex equine inhaler device

Single doses of one, three, and six actuations (120 micro g albuterol/actuation) and multiple daily doses (six actuations per dose four times daily) for 5 days of aerosol albuterol sulfate were sequentially administered to each of six horses using an equine inhaler device (Torpex, 3M Animal Care Products, St. Paul, MN [corrected] and Boehringer Ingleheim Vetmedica, Inc., St. Joseph, MO [corrected]). A 2-week washout period was allowed between each dose. ELISA testing revealed no evidence of albuterol in urine at 24 hours after any single-dose administration. Results indicated that 48 hours or longer should be allowed for albuterol to be cleared from urine after single doses. When given at the maximum recommended rate of six actuations per dose four times a day for 5 days, urine samples tested by ELISA showed no evidence of albuterol at 48 hours after the final dose. Testing of nasal swabs by ELISA demonstrated the presence of albuterol for 8 hours after each single dose, and some horses might have detectable levels of albuterol in nasal swabs for several days following administration of multiple doses. As a guideline for withdrawal time, 72 hours or longer should be allowed after administration of aerosol albuterol sulfate to horses before participation in equestrian competitions that are regulated for detection of certain performance-enhancing substances. However, these recommendations were based on a small sample of horses and the specific ELISA test used and interpreted as described. Factors specific to individual horses may influence these detection times.     

Shipping suppresses lymphocyte blastogenic responses in Angus and Brahman X Angus feeder calves

An experiment using 40 Angus or Brahman X Angus preconditioned feeder calves was conducted to evaluate the influence of shipping on cellular immune reactivity. Steers were allotted on the basis of weight and breed to a control or shipped group. Shipped steers were trucked 700 km to a feedlot; control steers remained at the ranch of origin. Total and differential leukocyte counts, phytohemagglutinin skin-test responses, lymphocyte blastogenic responses, monocyte phagocytic function, packed cell volumes and concentrations of plasma cortisol were determined before, immediately after and 1 wk after shipment. At unloading, total leukocytes were increased (P less than .05) in shipped Angus steers. Shipped steers also had higher (P less than .01) numbers of neutrophils. Skin-test responses to phytohemagglutinin were higher (P less than .05) in Angus than in Brahman X Angus steers, but shipping did not influence the reaction. Lymphocyte blastogenic responses were lower (P less than .05) in shipped steers; however, cortisol levels in plasma were not elevated (P greater than .10) in shipped calves. Monocyte phagocytosis and packed cell volume were not influenced by shipping. These data suggest that shipped steers have suppressed lymphocyte blastogenic responses.     

Managing America's wild horses and burros

Fifteen years after passage of the Wild Free-Roaming Horse and Burro Act, die Bureau of Land Management and the Forest Service had made significant progress in the management of this resource, but many aspects of the program remained controversial. Program emphasis centered on control of herd size as prescribed in locally developed resource management plansand disposition of excess animals that were captured and removed. Population trends from 1971 to 1986 indicated a steady increase in numbers through 1984 and a gradual decline since that time,but numbers were still about twice the estimated appropriate management level. Budget trends reflected an increased concern in the Congress about management levels and a rapidly escalating cost of maintaining unadopted excess animals. Proposed solutions to these problems included destruction of unadopted animals or their sale, the latter of which would require legislation. Research conducted on wild horses and burros through 1982 centered on range relationships, census techniques, and foaling rates. A new research effort called for in the 1985 appropriation focused on fertility control methods and strategies for population management. A wild horse and burroadvisory board was established in 1986 to advise the Secretaries of the Interior and Agriculture on matters pertaining to management, protection, and control of wild horses and burros on the Nation's public lands.     

Identification of lidocaine and its metabolites in post-administration equine urine by ELISA and MS/MS

Lidocaine is a local anesthetic drug that is widely used in equine medicine. It has the advantage of giving good local anesthesia and a longer duration of action than procaine. Although approved for use in horses in training by the American Association of Equine Practitioners (AAEP), lidocaine is also an Association of Racing Commissioners International (ARCI) Class 2 drug and its detection in forensic samples can result in significant penalties. Lidocaine was observed as a monoprotonated ion at m/z 235 by ESI+ MS/MS (electrospray ionization-positive ion mode) analysis. The base peak ion at m/z 86, representing the postulated methylenediethylamino fragment [CH2N(CH2CH3)2]+, was characteristic of lidocaine and 3-hydroxylidocaine in both ESI+ and EI (electron impact-positive ion mode) mass spectrometry. In addition, we identified an ion at m/z 427 as the principal parent ion of the ion at m/z 86, consistent with the presence of a protonated analog of 3-hydroxylidocaine-glucuronide. We also sought to establish post-administration ELISA-based 'detection times' for lidocaine and lidocaine-related compounds in urine following single subcutaneous injections of various doses (10, 40, 400 mg). Our findings suggest relatively long ELISA based 'detection times' for lidocaine following higher doses of this drug.     

New therapeutic approaches for equine protozoal myeloencephalitis: pharmacokinetics of diclazuril sodium salts in horses

Diclazuril is a triazine-based antiprotozoal agent which may have clinical application in the treatment of equine protozoal myeloencephalomyelitis (EPM). In this study, the use of the sodium salt diclazuril to increase the apparent bioavailability of diclazuril for the treatment and prophylaxis of EPM and various other Apicomplexan mediated diseases is described. In this study, diclazuril sodium salt was synthesized and administered to horses as diclazuril sodium salt formulations. The absorption, distribution, and clearance of diclazuril sodium salt in the horse are described. Diclazuril was rapidly absorbed, with peak plasma concentrations occurring at 8-24 hours following an oral mucosal administration of diclazuril sodium salt. The mean oral bioavailability of diclazuril as Clinacox was 9.5% relative to oral mucosal administration of diclazuril sodium salt. Additionally, diclazuril in DMSO administered orally was 50% less bioavailable than diclazuril sodium salt following an oral mucosal administration. It was also shown that diclazuril sodium salt has the potential to be used as a feed additive for the treatment and prophylaxis of EPM and various other Apicomplexan mediated diseases.