Validation and comparison of two methods of measuring lactate in equine plasma.

REASONS FOR PERFORMING STUDY: Some methods of lactate (LA) measurement have not been validated appropriately for use in horses. OBJECTIVES: To validate 2 LA analysers (YSI 2300 Stat Plus and TDx Lactic Acid Assay) for use with equine plasma and to compare plasma [LA] determined by the 2 methods. METHODS: Both instruments were evaluated for linearity, parallelism, recovery and precision using serial dilutions of standard LA solutions and equine plasma and then comparing results with linear regression or paired t tests. Plasma [LA] results were compared in 275 blood samples collected from horses exercising at various intensities using Bland-Altman analysis. Level of significance was P < 0.05. RESULTS: YSI exhibited good linearity for both LA standards and equine plasma (P < 0.05) at 0-30 mmol/l. TDx had good linearity at 0-12 mmol/l (P < 0.05); with LA standard solutions >12 mmol/l and with equine plasma, linearity was decreased. YSI exhibited good parallelism between LA standards and equine plasma LA measurements throughout the 0-30 mmol/l range (P > 0.05). Parallelism was poor with TDx (P < 0.05). Mean ± s.d. % recovery was 101.7 ± 3.4% for YSI (acceptable) and 110.6 ± 8.4% for TDx (unacceptable). Within-run and mean between-run coefficients of variation (CV) for plasma samples tested from 3.3-29.5 mmol/l were 0.4-3.0% for YSI. CVs for samples tested from 2.8-8.0 mmol/l were 17.4-24.1% for TDx. In 275 plasma samples, [LA] ranged from 0.1-42.7 mmol/l and 0.3-50.6 mmol/l for the YSI and TDx methods, respectively. The difference in plasma [LA] determined by the 2 methods was -1.0 ± 3.2 mmol/l, documenting that the TDx overestimated the YSI results by a mean value of 1 mmol/l. CONCLUSIONS: It was concluded that the YSI method was a reliable method for measuring equine plasma [LA] from 0-30 mmol/l. The TDx method was found not to be suitable for use with equine plasma due to greater variability in measurements (high CV).

Technical note: using calcium carbonate as an osmolar control treatment for acid-base studies in horses.

The efficacy of using calcium carbonate as an osmolar control treatment for acid-base studies in horses receiving alkalizing compounds was evaluated. Six mares were nasogastrically intubated with isomolar quantities of sodium or calcium as sodium bicarbonate or calcium carbonate or with water during three treatment periods. Doses of the carbonic acid salts were 500 mg/kg sodium bicarbonate mixed with 4 L of distilled water (positive control) and 595 mg/kg calcium carbonate mixed with 2 L of distilled water to yield isoosmolar treatments. Four liters of distilled water served as the negative control. Jugular venous blood samples were drawn before intubation and at hourly intervals for 6 h after intubation. The serum electrolytes Na+ and K+, blood pH, and HCO3- were determined. The sodium bicarbonate treatment increased blood pH and HCO3- (P < 0.01) above both the water and CaCO3 treatments. No differences (P > 0.05) were found between the water and CaCO3 treatments. These data indicate that calcium carbonate may serve as a suitable osmolar control treatment for studying the effects of treatments that affect acid-base status of horses.

Dietary fat supplementation effects on in vitro nutrient disappearance and in vivo nutrient intake and total tract digestibility by horses.

Addition of fat to the diet of the equine is a popular method of increasing energy density of the diet while reducing feed intake. Reducing feed intake is of interest to race horse trainers because additional feed is seen as additional weight and, therefore, a hindrance to performance. Limited information is available regarding the interactions of fat with other dietary components, particularly fiber, in the equine digestive system. The effect of dietary fat on in vitro nutrient disappearance in equine cecal fluid was studied in Exp. 1 using a split-plot design within a 2 x 2 Latin square. Two ponies were fed alfalfa (ALF) alone or alfalfa plus 100 g/d corn oil. Five substrates were used to determine in vitro DM disappearance, OM disappearance, NDF disappearance, and total dietary fiber (TDF) disappearance. The substrates included: ALF, tall fescue (TF), red clover (RC), soybean hulls (SBH), and rolled oats (RO). Fat supplementation did not affect in vitro DM, OM, or NDF disappearance. Addition of fat to the diet increased (P < 0.05) the disappearance of NDF in RO. Among substrates, in vitro DM and OM disappearance were highest (P < 0.05) for RO, followed by SBH, ALF, RC, and TF. In vitro NDF and TDF disappearance were highest (P < 0.05) for SBH, followed by RO, ALF, RC, and TF. In Exp. 2, the effects of varying levels of fat on nutrient intake and total tract digestibility were examined using a 4 x 4 Latin square design. Four mature mares were fed a 60% forage-40% concentrate diet containing different concentrations of fat: 0% supplemental fat control (C); 5% supplemental corn oil (5% CO); 10% supplemental corn oil (10% CO); or 15% supplemental corn oil (15% CO). Treatment did not affect intake of the concentrate portion of the diet or CP, gross energy, or NDF intake. Mares consuming the C diet had the highest (P < 0.05) intake of alfalfa cubes, DM, and OM, followed by those on the 10, 5, and 15% CO treatments, respectively. Treatment did not affect nutrient digestibility. Mares consuming the 15% CO diet had the highest (P < 0.05) fat digestibility, whereas those consuming C had the lowest fat digestibility. Fat in the form of CO generally had little effect on in vitro and in vivo nutrient digestibilities in horses.

Hemodynamic effects of thyroidectomy in sedentary horses.

OBJECTIVE: To investigate hemodynamic effects of thyroidectomy in horses at rest. ANIMALS: 6 healthy aged Quarter Horse mares. PROCEDURE: Horses were monitored for 5 months before and 4 weeks after thyroidectomy and for an additional 4 weeks after administration of thyroid hormone supplement (2.5 microg of thyroxine/kg of body weight, PO, q 12 h, and 0.6 microg of triiodothyronine/kg, PO, q 12 h). Responses to thyroid-stimulating hormone (TSH) were measured before and 4 weeks after thyroidectomy. Other variables monitored daily were resting rectal temperature (T), heart rate (HR), respiratory rate (RR), and body weight (BW). Monthly cardiac output (Q), blood volume (BV), plasma volume (PV), standard electrocardiographic measures, systolic and right ventricular blood pressure, and HR responses were determined after IV administration of isoproterenol and phenylephrine. Variables were analyzed by use of repeated-measures ANOVA. RESULTS: Complete thyroidectomy was confirmed by minimal response to TSH 4 weeks after surgery. Resting HR, RR, T, Q, and beta-adrenergic responsiveness to isoproterenol decreased significantly after thyroidectomy. Resting T, Q, and beta-adrenergic responsiveness increased after administration of supplement and was not significantly different from euthyroid values. Blood volume and PV increased significantly after thyroidectomy but did not return to euthyroid values despite administration of supplement. Response to phenylephrine was minimally different between treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Thyroidectomy in horses caused decreased resting HR, RR, T, Q, and isoproterenol responsiveness and increased BV, PV, PQ interval, and QT interval corrected for HR. Some of these surgically induced changes appeared to be partially reversed by administration of thyroid hormone supplement.

Effects of prerace exercise, frusemide, sex and ambient temperature on blood sodium, bicarbonate and pH values in standardbred horses.

Analysis of data collected at racetracks showed that temperature, the diuretic drug, frusemide, exercise, temperature/exercise interaction and sex/age had significant (P < 0.05) effects on pH and bicarbonate ion concentration (P < 0.01). Sodium concentrations were significantly (P < 0.001) affected by temperature and frusemide. We suggest that the normal range limits for blood sodium, bicarbonate and pH used in prerace testing procedures should be adjusted for ambient temperature and for horses given frusemide and/or prerace exercise. These adjustments should improve the precision of detecting illegal alkalogenic agents and incidence of false positive and false negative test results.

Measurements of heart scores and heart weights in horses of two different morphic body types.

Heart scores (the arithmetic mean of the QRS durations of the three bipolar limb leads for 10 cycles) and heart weights were determined from five draft type horses and five racing type horses, and evaluated for relationships between heart score and heart size within and between the two different types of horses. Horses were killed using a captive bolt gun, and actual heart weights were recorded using an electronic digital scale. The linear regression equation for heart weight versus heart score had a significant (P < 0.05) negative slope for racing types. The regression coefficients for heart weight versus heart score for draft types were not significantly (P > 0.05) different than zero. There was no significant (P > 0.05) difference found between draft and racing types. Significant (P < 0.01) differences were found between the heart ratios (heart weight as a percentage of live body weight) of racing and draft types. Heart scores were not found to be significantly (P > 0.05) different between racing and draft type horses.

Changes in the metabolic profile of equine muscle from birth through 1 yr of age.

The purpose of this study was to investigate metabolic changes in equine muscle from birth to 1 yr of age. Duplicate biopsies from the middle portion of the gluteus medius were obtained from a depth of 2 cm beneath the superficial fascia at 1 day, 7 days, 1 mo, 3 mo, 6 mo, and 1 yr of age in 11 quarter horses and at 1 day, 3 mo, 6 mo, and 1 yr of age in 5 Standardbreds. Muscle enzyme activities determined were citrate synthase, 3-hydroxyacyl-CoA dehydrogenase, phosphorylase, and lactate dehydrogenase. Percent fast-twitch, fast-twitch high oxidative, and slow-twitch oxidative fiber types were determined using succinate dehydrogenase and myosin adenosinetriphosphatase (pH 9.4) histochemical stains. Histochemically determined muscle fiber-type percents did not change dramatically with increasing age. However, lactate dehydrogenase activity increased threefold in quarter horses and twofold in Standardbreds, and phosphorylase activity increased sixfold in quarter horses and sevenfold in Standardbreds from 1 day to 6 mo of age. Citrate synthase and 3-hydroxyacyl-CoA dehydrogenase activities decreased during the first 3 mo of age in quarter horses.

Changes in the metabolic profile of the equine gluteus medius as a function of sampling depth.

1. Cross sections from the middle of the gluteus medius were removed from 10 adult horses and used to evaluate changes in histochemically determined muscle fiber type and biochemically determined metabolic enzyme activities as a function of sample depth. 2. Muscle fiber types determined using histochemical methods for myosin ATPase (pH 9.4) and succinic dehydrogenase (SDH) activity indicated percent fast-twitch glycolytic (FG) muscle fibers decreased and slow-twitch oxidative (SO) fibers increased as a function of increasing sampling depth. 3. Percent histochemically determined fast-twitch oxidative glycolytic (FOG) fibers decreased slightly only in the deepest region of the gluteus medius. 4. Citrate synthase (CS) enzymatic activity, used as a marker for mitochondrial oxidative potential, increased 2.5-fold in activity per g of muscle protein from 1 to 8 cm sampling depth. 5. 3-hydroxyacyl-CoA dehydrogenase (HAD) enzymatic activity, used as a marker for lipid oxidation potential, increased 3-fold in activity per g of muscle protein when the depth increased from 1 to 8 cm. 6. Phosphorylase (PS) enzymatic activity, used as a marker for potential glycogen utilization, decreased 50% in activity per g of muscle protein when going from 1 to 8 cm. 7. Lactate dehydrogenase (LDH) enzymatic activity, used as a marker for anaerobic glycolytic potential, decreased about 50% in activity as the sampling depth increased from 1 to 8 cm. 8. In summary, the superficial portion of the equine gluteus medius was found to be more glycolytic and less aerobic in its metabolic profile than deeper regions. The muscle became progressively more aerobic and less glycolytic with increasing sampling depth.