Plasma firocoxib concentrations after intra-articular injection of autologous conditioned serum prepared from firocoxib positive horses.

Orthobiologics such as autologous conditioned serum (ACS) are often used to treat joint disease in horses. Because ACS is generated from the horse's own blood, any medication administered at the time of preparation would likely be present in stored ACS, which could lead to an inadvertent positive drug test following intra-articular (IA) injection. The main objective of this study was to determine if ACS prepared from firocoxib positive horses could result in detectable plasma concentrations of the drug following IA injection. Firocoxib was administered to six horses at 0.1mg/kg PO twice at a 24h interval. Blood was obtained at 4h following the second dose and transferred to a separate syringe (Arthrex IRAP II) for ACS preparation. Plasma and ACS concentrations of firocoxib were analysed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). When horses were confirmed firocoxib negative, 7.5mL of ACS was injected into both tarsocrural joints. Blood samples were collected at 0, 4, 8, 12, 24, and 48h, and firocoxib concentration was measured. Mean (±standard error of the mean, SEM) plasma concentration of firocoxib 4h following the second dose was 33.3±4.72ng/mL. Mean (±SEM) firocoxib concentration in ACS was 35.4±4.47ng/mL. Fourteen days following the second and last dose of firocoxib, mean plasma concentration was below the lower limit of detection (LOD=1ng/mL) in all horses. Following IA injection of ACS, plasma concentrations of firocoxib remained below LOD at all times in all horses. ACS generated from horses with therapeutic plasma concentrations of firocoxib did not contain sufficient firocoxib to lead to a positive plasma drug test following IA administration.

Absence of cadmium in the blood of horses fed oats grown on municipal sludge-amended soil.

Effluents from a number of industries which are typically treated in municipal sewage plants result in cadmium-containing sludge. Disposal of such sewage sludge by application to agricultural land can result in uptake of cadmium by crops. In this study, oats were grown on soils which had been amended with sewage sludge from Syracuse, New York. The cadmium concentration in the sludge-grown oats was 1.79 ppm dry weight. Horses were fed the sludge-grown oats for 6 weeks during which time blood samples were taken for the determination of cadmium. No significant differences (p greater than 0.05) were found in the concentration of cadmium in the blood of the horses consuming the oats compared with that in the blood before feeding began (control). The well-known efficient accumulation of renal cadmium by horses may account for a lack of significantly increased cadmium in blood during the feeding period.

Qualitative and quantitative analysis of hydrochlorothiazide in equine plasma and urine by high-performance liquid chromatography.

A sensitive, quantitative method has been developed for the determination of hydrochlorothiazide in equine plasma and urine. Thin-layer chromatography is used to screen for the presence of the drug in unknown samples. The TLC screening methods described provide minimum detection limits of 50 ng/mL in plasma and 25 ng/mL in urine. A silica micro chromatography column is used to clean up ethyl acetate extracts for HPLC analysis and mass spectral confirmation. An internal standard, trichloromethiazide, is used to derive quantitative data at concentrations as low as 25 ng/mL for plasma disappearance curves and urinary excretion rates.

Changes in articular cartilage after intra-articular injections of methylprednisolone acetate in horses.

Eight mature horses with no prior signs of joint disease or history of intra-articular therapy were treated with 8 weekly intra-articular injections of methylprednisolone acetate. Treatments were given at a dose of 120 mg/joint into the right radiocarpal and intercarpal joints, with the left joints as untreated controls. Articular cartilage samples were obtained at necropsy 1, 4, and 8 weeks after the last injection. Compared with controls, cartilage from injected joints had a loss of hematoxylin basophilia and decreased intensity of staining in safranin O fast green dye. Chondrocyte necrosis and hypocellularity were observed in all samples of cartilage from treated joints. Proteoglycan content and its rate of synthesis were reduced. There was a progressive loss of proteoglycan content, whereas proteoglycan synthesis increased somewhat 4 and 8 weeks after treatment. Collagen content was unchanged, but its rate of synthesis was markedly inhibited. Collagen synthesis did not recover, but remained decreased at 5 to 15% of the values from untreated cartilage. Water percentage was increased, but fibronectin content was not significantly different. A single injection of methylprednisolone acetate was also given into the right metacarpophalangeal joints of 3 of the 8 horses in this group, with the left joints serving as untreated controls. Sixteen weeks after the treatment, cartilage of the treated joints had a loss of histochemical staining and proteoglycan content was reduced to 50% of control values. The mean rate of proteoglycan synthesis and mean fibronectin content were increased, but the differences were not statistically significant (P greater than 0.05). Other variables were essentially unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)

Toxic and protective constituents in pet foods.

An analytical survey of mutagens, nitrosamines, polychlorinated biphenyls, toxic elements, and gamma-emission, as well as the toxicologically protective constituents zinc, selenium, and vitamin C, in 48 pet foods was conducted. Aside from high concentrations of fluoride and iodide in some samples and the expectedly higher concentrations of mercury and selenium in certain cat foods containing fish, the samples were notably free of the other toxic constituents. Direct-acting and promutagens and nitrosamines were not detectable in any of the samples. gamma-Emission was very low in all of the foods. Polychlorinated biphenyls were only detected in one cat food.

Identification of detomidine carboxylic acid as the major urinary metabolite of detomidine in the horse.

Horse urine was investigated for metabolites by chromatography and mass spectrometry following the oral administration of the large animal analgesic sedative detomidine to two stallions and intravenous administration of [3H]-detomidine to a mare. Detomidine carboxylic acid and hydroxydetomidine glucuronic acid conjugate were identified in the urine after the oral doses. In addition, traces of free hydroxydetomidine were observed. About half of the radioactivity of [3H]-detomidine was excreted in the urine in 12 h after the i.v. dose (80 micrograms/kg). Most of the excretion occurred between 5 and 12 h in contrast to urine output which was highest 2-5 h after the dosing. The major radioactive metabolite in the urine was detomidine carboxylic acid. It comprised more than two thirds of the total metabolites in all the urine fractions collected. Its excretion profile was similar to that of total radioactivity. Hydroxydetomidine glucuronide was also excreted. It contributed 10-20% of the total metabolites in the urine. The free aglycone was only seen in the samples collected during the peak urine flow. A minor metabolite was tentatively characterized as the glucuronide of N-hydroxydetomidine.

Low dose exogenous erythropoietin elicits an ergogenic effect in standardbred horses.

REASONS FOR PERFORMING STUDY: Recombinant human erythropoietin (rhuEPO) causes an increase in red blood cell production and aerobic capacity in other species; however, data are lacking on effects in the horse. HYPOTHESIS: This study tested the hypothesis that rhuEPO administration would alter red cell volume (RCV), aerobic capacity (VO2max) and indices of anaerobic power. METHODS: Eight healthy, unfit mares accustomed to the laboratory and experimental protocols were randomly assigned to either a control (CON, n = 4; 3 ml saline 3 times/week for 3 weeks) or EPO group (EPO, n = 4, 50 iu/kg bwt rhuEPO/3 ml saline 3 times/week for 3 weeks). Exercise tests (GXT) were performed on a treadmill (6% incline), 1 week before and 1 week after treatment. The GXT started at 4 m/sec, with a 1 m/sec increase every 60 sec until the horse reached fatigue. Oxygen uptake was measured via an open flow indirect calorimeter. Blood samples were collected before, during (each step) and 2 and 15 min post GXT to measure packed cell volume (PCV), haemoglobin concentration (Hb), blood lactate concentration (LA) and plasma protein concentration (TP). Plasma volume (PV) was measured using Evans Blue dye. Blood volume (BV) and RCV were calculated using PCV from the 8 m/sec step of the GXT. RESULTS: There were no alterations (P>0.05) in any parameters in CON horses. By week 3, EPO produced increases (P<0.05) in resting PCV (37 +/- 2 vs. 51 +/- 2) and Hb (37%). RCV (26%) and VO2max (19%) increased, but BV did not change (P>0.05) due to decreased PV (-11%, P<0.05). There was a significant increase in velocity at VO2max and LApeak for horses treated with rhuEPO and substantial decrease (P<0.05) in VO2 recovery time when the pretreatment GXT was compared to the post treatment GXT. No differences (P<0.05) were detected for TP, VLA4, run time or Vmax. CONCLUSIONS: Low dose rhuEPO administration increases RCV and aerobic capacity without altering anaerobic power. POTENTIAL RELEVANCE: This study demonstrates that rhuEPO enhances aerobic capacity and exercise performance, a question relevant to racing authorities.

A simple method for detecting ethylene glycol in urine by thin layer chromatography.

A thin layer chromatographic method is presented for determining ethylene glycol in urine. The method uses 5% vanillin in sulfuric acid as the chromogenic reagent. Since the method is simple and sensitive, it provides a practical means of diagnosing ethylene glycol poisoning in clinical situations.

Analysis of propionylpromazine and its metabolites in horse urine.

The metabolism of propionylpromazine in the horse was studied. Although propionylpromazine is not currently approved or recommended for use in horses, it has been used illegally to alter their performance. Propionylpromazine hydrochloride was administered intramuscularly at clinical and subclinical doses. Three metabolites were detected in urine. The major metabolite was identified as 2-(1-hydroxypropyl) promazine sulfoxide. The detection of this metabolite in routine drug testing has been described.

Industrial fluoride pollution. Chronic fluoride poisoning in Cornwall Island cattle.

An aluminum plant on the south bank of the St. Lawrence river, southwest of Cornwall Island, Ontario, Canada, has emitted 0.816 metric tons of fluoride daily since 1973; considerably higher amounts were emitted from 1959 to 1973. The plant has been designated as the "major source of fluoride emissions impacting on Cornwall Island." Chronic fluoride poisoning in Cornwall island cattle was manifested clinically by stunted growth and dental fluorosis to a degree of severe interference with drinking and mastication. Cows died at or were slaughtered after the third pregnancy. The deterioration of cows did not allow further pregnancies. Fluoride concentrations in ash of biopsied coccygeal vertebrae increased significantly with age and were dependent on distance from and direction to the aluminum plant. Fluoride in bone ash of a 7-month old-fetus exceeded 500 ppm; fluoride thus was passed transplacentally. Analyses of fluoride in ash of bones obtained at necropsy of cattle from 4 months of age to 4 to 5 years of age showed increased amounts with age. Cancellous bone retained far higher amounts than cortical bone, a reflection of the normally higher metabolic rate of cancellous bone. Concentrations exceeding 10,000 ppm fluoride were recorded in cancellous bone of a 4-to 5-year-old cow. The target cells for fluoride in chronic fluorosis were shown to be the ameloblasts, the dental pulp cells and the odontoblasts and, in bone, primarily the resorbing osteocytes and also the osteoblasts. Atrophy and necrosis of the ameloblasts were responsible for enamel defects. The existing enamel showed brown discoloration from fluoride deposits. The pulp cells underwent fibrous and osseous metaplasia and necrosis of the ectopic bone occurred. The odontoblasts were atrophic and the dentin showed brown discoloration. The resorbing osteocytes were inactive and osteosclerosis resulted. This was especially pronounced in areas of normally great apposition, i.e. in the metaphyses. The epiphyseal plate became squeezed between petrotic bone and growth was stunted. Resorption of alveolar bone surrounding the deciduous teeth was severely retarded or arrested. A delay in eruption of permanent teeth occurred; it was up to 3.5 years in incisor teeth. Interference with the resorbing osteocytes in fluorotic bone was also demonstrated by loss of collagen birefringency in such bone. Failure of bone resorption also caused retention of trabecular bone in the cortices; this was observed even in a 4-t0-5-year-old cow. In areas where modeling into osteonic bone had begun, fluoride deposits were extremely heavy but this bone showed numerous soft osteons in microradiographs. The toxic effect of fluoride on osteocytes also resulted in the death of the cells. Such osteonecrosis occurred mainly in gnathic bone. There was atrophy of the osteoblasts. Osteopenia thus resulted from osteonecrosis and osteoporosis. Subperiosteal exostoses were not observed in long bones. The degree of fluorosis in Cornwall Island cattle was severe...

The extraction and isolation of dexamethasone-related compounds.

Radiolabeled dexamethasone (9-fluoro-16 alpha-methyl-11 beta, 17, 21-trihydroxy-1,4-pregnadiene-3,20-dione) was utilized to develop a practical and relatively expedient method of detection for dexamethasone and its major metabolite in horse urine. The effects of solvent, pH, salt saturation, and back extraction on the extraction efficiency of dexamethasone-related compounds and the presence of endogenous background were evaluated. Final isolations of dexamethasone and its major urinary metabolite were procured by successive thin-layer chromatography (TLC) in three different developing systems.

Milk production of cows exposed to industrial fluoride pollution.

Milk sales records for a dairy herd showed that milk production did not decrease during the first 4 yr of exposure of cows to fluoride pollution from an aluminum plant. During the next 3 yr milk production decreased, but not significantly. From yr 8 of exposure there was a significant decrease, which persisted through yr 19, when the dairy operation was terminated. From yr 15 through yr 19, the milk yield averaged less than 60% of the expected value.

Drug analysis by direct liquid introduction micro liquid chromatography mass spectrometry.

The analytical capabilities of a micro high performance liquid chromatograph interfaced to an unchanged quadrupole mass spectrometer are presented. Continuous monitoring of the total micro liquid chromatographic effluent allows full scan chemical ionization mass spectra of from one to five nanograms of drugs and their metabolites to be recorded. The interface is a simple, inexpensive device which can be assembled from commercially available components. An eight microliter per minute flow rate of the micro liquid chromatographic eluant allows separation and identification of biologically important substances not amenable to gas chromatography mass spectrometry techniques. The sensitivity of micro liquid chromatography mass spectrometry performed as described is comparable with gas chromatography mass spectrometry and is achieved by introducing the total micro liquid chromatographic effluent into the chemical ionization ion source of the mass spectrometer. Selected ion monitoring provides 20 pg detection limits of phenothiazine tranquilizers injected on column.

Vascular pathology in phenylbutazone intoxicated horses.

Three mature Thoroughbred geldings were given 13.63 mg phenylbutazone/Kg bodyweight intravenously for 3 days and repeated in one horse 4 days later. After 4, 7 and 10 days (double treatment), degeneration of the wall of small veins occurred in all horses. The veins were dilated and/or showed hyalin degeneration. The phlebopathy was interpreted to be paramount in phenylbutazone intoxication. All other manifestations, including erythro- and leukodiapedesis, submucosal edema and ulceration of the gastrointestinal mucosa, phlebothrombosis and significant changes in the hemogram and serum chemistry, were considered secondary to the vein lesions.

New York State and U.S. Federal fluoride pollution standards do not protect cattle health.

Fluoride emissions from an aluminum plant in New York State just west to the bridge to Cornwall, Ontario, Canada, are in compliance with New York State and U.S. Federal standards. Ambient air fluoride virtually never exceeds New York State standards. In a New York State dairy farm, downwind from the aluminum plant about 40% of the time and with the fields within 1300 to 2800 m from the plant, fluoride contamination of forage ranged from 13 to 25 ppm, well below the 40 ppm which is the "tolerance" level by National Academy of Sciences. Sixty-three of 82 dairy cattle on that farm were slaughtered in 1979 because of chronic fluoride poisoning. In the 19 cattle left on the farm in June, 1979, there was no dental fluorosis in calves less than 4 months of age, mild to moderate dental fluorosis in older calves and heifers and severe dental fluorosis in the 4 young adult cattle. Ash fluoride in a stillborn calf was 280 ppm and in the oldest cattle 2800; the increase was significantly correlated to age. It is concluded that New York State and U.S. Federal standards for fluoride emissions, New York State standards for ambient air fluoride and National Academy of Sciences "tolerance" levels for ingestion of fluoride do not protect cattle health.