Kenneth H. McKeever

Exercise training-induced hypervolemia in the horse.

The purpose of this study was to determine if a chronic hypervolemia would accompany endurance exercise training in the horse. Six mature previously inactive horses were utilized for this study. During the 5-wk experiment, five of the horses were trained for 14 d on a treadmill ergometer at a constant treadmill speed of 5.6 km X hr-1 and a constant grade of 12.5% for graduated lengths of time. One horse was trained by lunging at a trotting pace in a round pen. Following training, plasma volume increased by 4.7 1 (29.1%, P less than 0.05). Although the rate of daily water intake did not change during the training period, 24-h urine output decreased by an average of 3.5 1 X d-1 (-24.5%, P less than 0.05). Resting glomerular filtration rate and the rate of sodium clearance were not altered by training. However, urea, potassium, and osmotic clearance were decreased by training (P less than 0.05) while free water clearance was increased (P less than 0.05). Resting plasma aldosterone and arginine vasopressin concentrations were not altered by training. Plasma potassium concentration was significantly decreased (P less than 0.05) following the 2 wk of training. These data would appear to suggest that renal control mechanisms affecting water reabsorption via the re-absorption of urea and osmotically active substances other than sodium provide the primary route for the training-induced hypervolemia seen in horses.

Chronic clenbuterol administration negatively alters cardiac function.

PURPOSE Chronic administration of pharmacological levels of beta2-agonists have been shown to have toxic effects on the heart; however, no data exist on cardiac function after chronic clenbuterol administration. The purpose of this study was to examine the effect of therapeutic levels of clenbuterol on cardiac performance. METHODS Twenty unfit Standardbred mares were divided into four experimental groups: clenbuterol (2.4 microg.kg(-1) twice daily 5 d.wk(-1)) plus exercise (20 min at 50% .VO(2max)) (CLENEX; N = 6), clenbuterol (CLEN; N = 6), exercise (EX; N = 4), and control (CON; N = 4). M-mode and two-dimensional echocardiography (2.5-MHz sector scanner transducer) were used to measure cardiac size and function before and immediately after an incremental exercise test, before and after 8 wk of drug and/or exercise treatments. RESULTS After treatment, CLENEX and CLEN demonstrated significantly higher left ventricular internal dimension (LVD) at end diastole (+23.7 +/- 4.8%; +25.6 +/- 4.1%), LVD at end systole (+29.2 +/- 8.7%; +40.1 +/- 7.9%), interventricular septal wall thickness (IVS) at end diastole (+28.9 +/- 11.0%; +30.7 +/- 7.0%), IVS at end systole (+29.2 +/- 8.7%; +40.1 +/- 7.9%), and left ventricular posterior wall systolic thickness (+43.1 +/- 14.%; +45.8 +/- 14.1%). CLENEX and CLEN had significantly increased aortic root dimensions (+29.9 +/- 6.1%; +24.0 +/- 1.7%), suggesting increased risk of aortic rupture. CONCLUSION Taken together, these data indicate that chronic clenbuterol administration may negatively alter cardiac function.

The effects of theaflavin-enriched black tea extract on muscle soreness, oxidative stress, inflammation, and endocrine responses to acute anaerobic interval training: a randomized, double-blind, crossover study

BackgroundMuscle soreness and decreased performance often follow a bout of high-intensity exercise. By reducing these effects, an athlete can train more frequently and increase long-term performance. The purpose of this study is to examine whether a high-potency, black tea extract (BTE) alters the delayed onset muscle soreness (DOMS), oxidative stress, inflammation, and cortisol (CORT) responses to high-intensity anaerobic exercise.MethodsCollege-age males (N = 18) with 1+ yrs of weight training experience completed a double-blind, placebo-controlled, crossover study. Subjects consumed the BTE (1,760 mg BTE·d-1) or placebo (PLA) for 9 days. Each subject completed two testing sessions (T1 & T2), which occurred on day 7 of the intervention. T1 & T2 consisted of a 30 s Wingate Test plus eight 10 s intervals. Blood samples were obtained before, 0, 30 & 60 min following the interval sessions and were used to analyze the total to oxidized glutathione ratio (GSH:GSSG), 8-isoprostane (8-iso), CORT, and interleukin 6 (IL-6) secretion. DOMS was recorded at 24 & 48 h post-test using a visual analog scale while BTE or PLA continued to be administered. Significance was set at P < 0.05.ResultsCompared to PLA, BTE produced significantly higher average peak power (P = 0.013) and higher average mean power (P = 0.067) across nine WAnT intervals. BTE produced significantly lower DOMS compared to PLA at 24 h post test (P < 0.001) and 48 h post test (P < 0.001). Compared to PLA, BTE had a slightly higher GSH:GSSG ratio at baseline which became significantly higher at 30 and 60 min post test (P < 0.002). AUC analysis revealed BTE to elicit significantly lower GSSG secretion (P = 0.009), significantly higher GSH:GSSG ratio (P = 0.001), and lower CORT secretion (P = 0.078) than PLA. AUC analysis did not reveal a significant difference in total IL-6 response (P = 0.145) between conditions.ConclusionsConsumption of theaflavin-enriched black tea extract led to improved recovery and a reduction in oxidative stress and DOMS responses to acute anaerobic intervals. An improved rate of recovery can benefit all individuals engaging in high intensity, anaerobic exercise as it facilitates increased frequency of exercise.

The endocrine system and the challenge of exercise

Fine-tuning of the response to exercise that lasts longer than a few seconds is reliant on the regulation of several key variables governing the cardiopulmonary, vascular, and metabolic response to exercise. This type of integrative response requires communication between organ systems that relies on the secretion of endocrine and paracrine substances by one tissue or organ that are transported remotely to other tissues or organs to evoke a response to adjust to the disturbance.

Exercise‐induced increases in inflammatory cytokines in muscle and blood of horses

REASONS FOR PERFORMING STUDY Studies have demonstrated increases in mRNA expression for inflammatory cytokines following exercise in horses and have suggested those markers of inflammation may play a role in delayed onset muscle soreness. However, measurement of mRNA expression in white blood cells is an indirect method. No studies to date have documented the cytokine response to exercise directly in muscle in horses. HYPOTHESIS This study tested the hypothesis that exercise increases cytokine markers of inflammation in blood and muscle. METHODS Blood and muscle biopsies were obtained from 4 healthy, unfit Standardbred mares (∼ 500 kg). The randomised crossover experiment was performed with the investigators performing the analysis blind to the treatment. Each horse underwent either incremental exercise test (GXT) or standing parallel control with the trials performed one month apart. During the GXT horses ran on a treadmill (1 m/s increases each min until fatigue, 6% grade). Blood and muscle biopsies were obtained 30 min before exercise, immediately after exercise and at 0.5, 1, 2, 6 and 24 h post GXT or at matched time points during the parallel control trials. Samples were analysed using real time-PCR for measurement of mRNA expression of interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and interleukin-1 (IL-1). Data were analysed using t tests with the null hypothesis rejected when P < 0.10. RESULTS There were no changes (P > 0.10) in IL-1, IL-6, IFN-gamma or TNF-alpha during control. Exercise induced significant increases in IFN-gamma, IL1 and TNF-alpha in blood and significant increases in IFN-gamma, IL-6 and TNF-alpha in muscle. There were no significant changes in mRNA expression of IL-1 in muscle or IL-6 in blood following the GXT. These cytokine markers of inflammation all returned to preGXT levels by 24 h post GXT. CONCLUSION High intensity exercise results in a transient increase in the expression of inflammatory cytokines in muscle and blood.

Chronic recombinant equine somatotropin (eST)administration does not affect aerobic capacity or exercise performance in geriatric mares

The purpose of this experiment was to test the hypothesis that chronic (89 days) administration of recombinant equine somatotropin (eST) would increase aerobic capacity and improve exercise performance in old mares. Fifteen, healthy, unfit, aged (20-26 year old) mares were randomly assigned to a treatment (eST, 12.5 mg day-1 in 3 ml glycine/manitol buffer, s.c., n = 7) or control (vehicle, 3 ml day-1, s.c., n = 8) group. Aerobic capacity and exercise performance were measured using a standardized exercise test (SET) performed on a high speed treadmill. Tests were conducted before (-21 days), during (+43 days and +89 days) and after (+127 days) treatment. During the SET, resting data were collected and the horses then ran up a fixed 6% grade, starting at 4 m s-1, with a 1 m s-1 increase every 60 s (omitting 5 m s-1) until fatigue. Oxygen uptake (VO2) was measured using an open flow calorimeter and blood lactate concentration (LA) via a lactate analyser. Venous blood samples (10 ml) were collected at rest, during the last 10 s of each step of the SET, and after exercise and used to measure LA, plasma protein concentration (PP), hematocrit (HCT), and the plasma concentrations of creatine kinase (CK) and aspartamine transferase (AST). There were no differences (P > 0.05) in resting VO2, LA, TPP, or HCT due to treatment or test time. Furthermore, there were no differences (P > 0.05) in maximal oxygen uptake (VO2max), top run velocity, run time, watts at VO2max, velocity to produce a lactate of 4 mmoll-1 (VLA4), watts at VLA4, peak HCT or peak LA. Finally, there were no differences (P > 0.05) in resting or post-exercise CK or AST. These data indicate that chronic eST administration does not affect aerobic capacity or indices of exercise performance in unfit aged mares.

Effect of orange peel and black tea extracts on markers of performance and cytokine markers of inflammation in horses

This study tested the hypothesis that orange peel (O) and decaffeinated black tea (T) extracts would alter markers of exercise performance as well as exercise-induced mRNA expression for the inflammatory cytokines IL-6, TNFalpha and IFN-gamma. Nine healthy, unfit Standardbred mares (age: 10 ^ 4years, ,450kg) were assigned to three treatment groups in a randomized crossover design where each horse was administered one of the following; placebo (O; 2l water), black tea extract in water (T; 2l) or orange peel extract in water (W; 2l), via a nasogastric tube. One hour later the horses completed an incremental graded exercise test (GXT) on a treadmill at a fixed 6% grade with measurements and blood samples obtained at rest, at the end of each 1min step of the GXT and at 2 and 5min post-GXT. An additional set of blood samples for Polymerase Chain Reaction (PCR) measurements of mRNA was obtained before exercise and at 5 and 30min and 1, 2, 4 and 24h post-GXT. The GXTs were conducted between 0700 and 1200h not less than 7days apart. There were no differences (P . 0.05) in VO2max, respiratory exchange ratio, run time, velocity at VO2max, core body temperature, haematocrit, creatine kinase (CK), plasma lactate concentrations, HR, right ventricular pressure (RVP) or pulmonary artery pressure (PAP) across treatments. A major finding was that orange peel extract significantly reduced post-exercise VO2 recovery time (W ¼ 112 ^ 7, O ¼ 86 ^ 6, and T ¼ 120 ^ 11s). There was a significant difference in plasma total protein concentration (TP) in the O runs compared with water and T. TNF-alpha mRNA expression was lower in the T runs compared with water and O trials. IFN-gamma mRNA expression levels appeared to be lower in both the Tand O extract runs compared with the water trials. The mRNA expression of IL-6 was unaltered across treatment groups. These data suggest that orange peel and black tea extracts may modulate the cytokine responses to intense exercise. Orange peel extract reduced post-exercise recovery time and may potentially enhance the ability of horses to perform subsequent bouts of high-intensity exercise.

Myosin heavy chain profiles and body composition are different in old versus young Standardbred mares.

There are limited data on age-related changes in body composition or skeletal muscle in the horse. Therefore, the purpose of this study was to investigate any differences in muscle myosin heavy chain (MHC) and body composition associated with aging. Twenty-three young (4-8 years) and eight old (20+ years) unfit Standardbred mares were evaluated. Rump fat thickness was measured using B-mode ultrasound and per cent body fat (% fat) was calculated. Needle muscle biopsies were obtained from right gluteus medius muscle. MHC composition was determined via sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Three MHC isoforms were subsequently identified as type I, type IIA, and type IIX and quantified using a scanning and densometric system. There were no significant differences (p>0.05) between old and young mares in fat (%) (19.0+/-6.4 vs 20.5+/-5.4), fat mass (kg) (102.3+/-39.9 vs 106.9+/-37.1), or body weight (kg) (529.4+/-34.9 vs 512.7+/-57.7). However, the old mares had significantly (p<0.05) greater lean body mass than the young mares (427.1+/-24.5 vs 405.7+/-37.9). Aged mares had significantly (p<0.05) less type I (7.8+/-2.9% vs 12.1+/-4.4%) and IIA (27.8+/-7.1% vs 36.1+/-9.5%) fibres than the young group but more type IIX (64.6+/-4.7% vs 51.8+/-11.1%). The MHC data are consistent with the age-related changes seen in other species.

Age related decreases in thermoregulation and cardiovascular function in horses

REASON FOR PERFORMING STUDY Older horses have an increased risk of hyperthermia due to impaired cardiovascular function. While many studies have investigated thermoregulation in horses during exercise, none have investigated the effects of ageing. OBJECTIVE To test the hypothesis that there is a difference in thermoregulation during exercise and plasma volume (PV) in young and old horses. METHODS Study 1: 6 young (Y, 7.7 ± 0.5 years) and 5 old (O, 26.0 ± 0.8 years) unfit Standardbred mares (507 ± 11 kg, mean ± s.e.) ran on a treadmill (6% grade, velocity calculated to generate a work rate of 1625 watts) until core temperature reached 40 °C. Core (CT), skin (ST), rectal temperature (RT) and heart rate (HR) were measured every min until 10 min post exertion. Packed cell volume (HCT), lactate (LA) and plasma protein (TP) were measured in blood samples collected before, at 40 °C and every 5 min until 10 min post exercise. Sweat loss was estimated using bodyweight. Study 2: Plasma volume was measured in 26 young (8.2 ± 0.7 years) and 8 old (26.6 ± 0.7 years) Standardbred mares (515 ± 12 kg) using Evans Blue dye. Pre-exercise blood (rBV) and red cell (rRCV) volumes were calculated using PV and HCT. Data analysis utilised repeated measures ANOVA and t tests and data are expressed as mean ± s.e. RESULTS Old horses reached 40 °C faster (998 ± 113 vs. 1925 ± 259 s; P < 0.05) with a greater HR at 40 °C (184 ± 6 vs. 140 ± 5 beats/min; P < 0.05) and greater sweat losses (P < 0.05). Heart rate did not differ (P > 0.05) post exercise. Age did not alter (P > 0.05) CT, ST, RT, LA, HCT or TP. Plasma volume was greater in Y vs. O horses (P < 0.05, 28.5 ± 1.4 vs. 24.1 ± 1.6 l) as was rBV (41.3 ± 2.0 vs. 35.3 ± 2.3 l) and rRCV (13.3 ± 0.6 vs. 11.1 ± 0.8 l). CONCLUSION Ageing compromises the ability to handle the combined demand of exercise and thermoregulation in part due to decreased absolute pre-exercise PV.

Cross-reactivity between human erythropoietin antibody and horse erythropoietin

Erythropoietin (EPO) is the primary hormone of erythropoiesis. Administration of recombinant human erythropoietin (rhuEPO) to improve racing performance in the horse represents a new form of blood doping, which has been associated with increased mortality. While immunoassay kits have become plentiful, very few commercial hormone assays are made specifically for equine research. There is a strong degree of sequence homology reported for EPO among species, which has allowed antibodes designed for human EPO research to be used to determine EPO concentration in other species. The objective of the present study was to use Western blot analysis to determine whether the antibody to rhuEPO, provided in a commercial radioimmunoassay (RIA) kit, recognizes horse EPO. Western blot analysis of purified rhuEPO and horse plasma was conducted, using the polyclonal goat‐antihuman EPO antibody supplied in the Incstar EPO Trac RIA as the primary antibody. Immunoblot analysis revealed a major band at approximately 52 kDa for both rhuEPO and the horse plasma. Our results demonstrate that a human EPO antibody recognizes equine EPO. These findings show that the Incstar EPO Trac RIA hormone assay system can be used to measure equine EPO.