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INTRODUCTION: Adipose tissue plays complex role(s) in metabolic and endocrine control. To date, little work has been done in the horse regarding adipocytokines. PURPOSE: This study was conducted to determine whether therapeutic levels of chronic beta-agonist administration, exercise, or both could alter their concentrations. METHODS: A total of 23 standard-bred mares were divided into four experimental groups: clenbuterol (2.4 mug x kg[-1] bw twice daily for 8 wk) plus exercise (8 wk, 20 min x d[-1] at 50% VO[2max]; CLENEX; N = 6), clenbuterol only (CLEN; N = 6), exercise only (EX; N = 5), and control (CON; N = 6). Rump fat thickness was measured using B-mode ultrasound and percent body fat (%fat) was calculated. Plasma adiponectin and leptin concentrations were measured using radioimmunoassay (RIA). In the absence of purified equine adiponectin or leptin, results were expressed as human equivalents of immunoreactive adipocytokines. RESULTS: The change in plasma immunoreactive (ir)-adiponectin HE concentration was negatively correlated (r = -0.520; P = 0.01) to the change in fat mass and positively correlated (r = 0.446; P = 0.03) to the change in fat-free mass. The change in plasma ir-leptin HE concentration was positively correlated (r = 0.550; P = 0.02) to the change in fat mass and negatively correlated (r = -0.473; P < 0.05) to the change in fat-free mass. CONCLUSION: These data demonstrate that a chronic clenbuterol administration alters the concentrations of the adipocytokines adiponectin and leptin in horses. These changes may play a role in previously reported repartitioning effects of clenbuterol. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine if calcium propionate is an alkalizing agent in exercising Standardbreds and if it alters plasma glucose and serum insulin concentrations. This study used a randomized crossover design to test the hypotheses that calcium propionate alters total CO2 (tCO 2 ), Ca ++ , pH, strong ion difference (SID), glucose, and insulin in Standardbreds completing a simulated race test (SRT) on a high-speed equine treadmill. Blood was collected from eight horses (mean age ± SD = 16 ± 2.7 years; range = 13 - 21 years) at 10 and 5 min prior to treatment or control administration, just prior to the subsequent SRT, directly after the SRT, and at 60- and 90-min post SRT. Plasma pH and plasma concentrations of tCO 2, glucose, Ca ++ , and Na + , K + , Cl - , Lac - (for SID calculation) were measured in duplicate by blood gas analyzer and serum insulin by radioimmunoassay. Data were analyzed by two-way repeated-measures ANOVA. For plasma pH, Na + , K + , Cl - , Lac - , SID, insulin and glucose, no effects of treatment or treatment by sampling-timepoint interaction were found (p>0.05). Plasma tCO 2 , Ca ++ , pH, Na + , SID, and glucose concentrations were significantly lower, and Lac - was significantly higher, directly after the SRT compared to all other timepoints (p<0.05). Plasma tCO2, HCO 3 - and Ca ++ were significantly higher in treated than in control horses at multiple sampling timepoints after treatment/control administration (p<0.05). Serum insulin concentration, measured only pre and 30 min post treatment/control administration, was unaffected by treatment. Calcium propionate is an alkalizing agent in horses., Competing Interests: Declaration of competing interest None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of this article., (Copyright © 2024. Published by Elsevier Inc.)

Introduction: Equine-assisted services (EAS) has received attention as a potential treatment strategy for post-traumatic stress disorder (PTSD), as existing literature indicates that symptoms may decrease following EAS. Relatively little is known about the mechanisms at play during lessons and if physiological measures are impacted. The objectives of this pilot study were to 1) explore the effects of adaptive horsemanship (AH) lessons on symptoms of PTSD, hormone concentrations, and social motor synchrony; 2) determine if physiological changes occur as veterans interact with horses; and 3) explore if the interaction between veteran and horse changes over the 8-week session., Methods: Veterans with PTSD were randomly assigned to control (CON, n = 3) or AH (n = 6) groups for an 8-week period (clinical trial; NCT04850573; clinicaltrials.gov). Veterans completed the PTSD Checklist (PCL-5) and Brief Symptom Inventory (BSI) at pre-, post-, and 2- and 6-month follow-up time points. They also completed a social motor synchrony test (pendulum swinging) and blood draw at pre- and post-time points. In weeks 1, 4, and 8, blood samples were drawn at 0 min, 3 min, 5 min, 25 min, and 30 min during the 30-min AH lessons. Veterans completed the Human-Animal Interaction Scale (HAIS) after each lesson. Blood samples were assayed for plasma cortisol, epinephrine, norepinephrine, and oxytocin. Data were analyzed with repeated measure ANOVAs. Changes in PTSD symptoms from pre- to post-time point were analyzed with paired t-tests., Results: Changes in PCL-5 scores tended to differ ( p = 0.0989), and global BSI scores differed ( p = 0.0266) between AH (-11.5 ± 5.5, mean ± SE; -0.5 ± 0.2) and CON (5.3 ± 5.4; 0.4 ± 0.2) groups. Social motor synchrony and hormone concentrations did not differ between groups or time points ( p > 0.05). Cortisol, norepinephrine, and oxytocin concentrations did not differ across sessions ( p > 0.05). Epinephrine concentrations tended ( p = 0.0744) to decrease from week 1 to 4 of sessions. HAIS scores increased ( p ≥ 0.0437) in week 3 and remained elevated as compared to week 1., Discussion: Participant recruitment was the greatest challenge. These preliminary results agree with the literature suggesting that EAS can reduce symptoms of PTSD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Rankins, Quinn, McKeever and Malinowski.)

Little literature exists on horses in adaptive horsemanship (AH) despite concerns about their well-being. The study objective was to evaluate behavioral and physiological responses of horses to ground-based AH lessons for veterans with post-traumatic stress disorder (PTSD). Lessons were expected to alter horses' hormone concentrations, behavior, and muscle activity. Geldings were assigned to AH (n=6; 20.3 ± 1.9 yrs., mean ± SE) or control (CON; stall in arena, n=6; 13.8 ± 1.7 yrs.) conditions for 8-week sessions based on current occupation (AH = equine-assisted services; CON = recreational riding). Plasma cortisol, epinephrine, norepinephrine, and oxytocin concentrations from samples at 0 (start of lesson), 3, 5, 25, and 30 (end) min were determined using assays validated in horses. Surface electromyography (sEMG) (masseter and brachiocephalic; Noraxon, Scottsdale, AZ, USA) and video were recorded continuously. Average rectified values (ARV) and median frequency (MF) were calculated (100 ms) after sEMG data were normalized, rectified, and filtered. The number, number of unique, and duration of stress related behaviors (ethogram) were recorded by three trained (ĸ ≥ 0.7) observers. Data were analyzed with repeated measures ANOVAs (significance P ≤ 0.05) with fixed effects of treatment, time point, week, and their interactions as appropriate and random effect of horse. CON horses had elevated cortisol concentrations (P = 0.0023) at 25 and 30 min. AH horses displayed fewer (P ≤ 0.0491) stress related and unique behaviors. CON horses were described as more (P < 0.0001) anxious, nervous, and stressed than AH horses (calm, comfortable, patient, and relaxed) in qualitative behavior analysis (22 observers). AH horses were less stressed than CON horses., Competing Interests: Declaration of competing interest None of the authors has any financial or personal relationships that could inappropriately influence or bias the content of the paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Equid welfare in equine assisted services (EAS) is an area that has received attention, but less attention than the documentation of human outcomes in response to EAS. To safeguard the well-being of equids and minimize human risk of injury, continued research on the effects of EAS programming and participants on equids needs to occur. The aims of this systematic scoping review were to identify the approaches taken for describing and understanding equids in EAS and the methods employed in evaluating equids' responses to EAS programming, participants, or both. Literature searches were performed in relevant databases to identify titles and abstracts for screening. Fifty-three articles were identified for full-text review. Fifty-one articles met the inclusion criteria and were retained for information and data extraction. The qualitative grouping of articles by study aim resulted in four categories: (1) characterization and description of equids in EAS; (2) the acute responses of equids to EAS programming, participants, or both; (3) the effects of management practices; and (4) the chronic responses of equids to EAS programming and participants. The latter three areas are in need of more research, especially as it relates to differentiating acute and chronic effects of EAS on the equids involved. Detailed reporting of information on study design, programming and participant characteristics, equid demographics, and workload are needed to facilitate comparison among studies and permit eventual meta-analysis of studies. Multi-faceted approaches including an array of measurements as well as relevant and informative control groups or conditions are required to identify the complex effects of EAS work on equids, their welfare, well-being, and affective states., Competing Interests: Declaration of Competing Interest There were no conflicts of concern when conducting this literature review., (Copyright © 2023 Elsevier Inc. All rights reserved.)

The purpose of this study was to examine the effects of an external nasal strip on thermoregulation during submaximal exercise in Standardbred horses. While several studies have been conducted to determine the effects of the external nasal strip on airway resistance, exercise induced pulmonary hemorrhage, gas exchange, and time to fatigue in maximally exercising horses, the effects of the nasal strip on equine thermoregulation have not yet been examined. It was hypothesized that the application of an external nasal strip would alter central venous temperature (T core ), skin temperature (T skin ), and exercise time to reach a central venous temperature of 40 °C. Eight mature Standardbred horses each performed two submaximal exercise trials, one with a nasal strip (NS), and one without (control), on a high-speed equine treadmill with exercise concluding upon T core reaching 40 °C. There were no significant differences in T core or T skin between the NS and control groups during pre-exercise, exercise or recovery (P > .05), nor were there differences (P > .05) in exercise time to reach a T core of 40 °C (NS: 11.8 ± 1.5 minutes; Control: 11.5 ± 1.1 min). We conclude that the application of an external nasal strip does not affect the equine thermoregulatory response during submaximal exercise., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Clenbuterol (β 2 agonist) is a commonly administered bronchodilator in race and performance horses. While long-term administration can alter exercise performance and muscle properties, little is known about its effects on these parameters following short-term administration. A single dose of clenbuterol (0.80 µg/kg) was expected to alter muscle activity of the extensor carpi radialis, semitendinosus, and longissimus dorsi during submaximal exercise. Eight mature Standardbred horses exercised for 2 minutes at 5 m/s on a high-speed treadmill following clenbuterol dosing (clenbuterol) or no dosing (control) in a crossover experimental design. Surface electromyography (sEMG) data were collected continuously from the muscles of interest and processed to determine average rectified value (ARV) and median frequency (MF) of the signal during peak muscle activation (100 ms period) during 15 strides. ARV data were log transformed. Data were analyzed with a mixed model ANOVA with fixed effects of period and treatment and a random effect of horse. No differences (P > .05) in amplitude (ARV) or frequency (MF) of the EMG signal were detected following clenbuterol administration. Thus, a one-time dose of clenbuterol had no statistically detectable effect on muscle activity during submaximal exercise. Further studies should be undertaken to confirm these results and examine the effects of long-term administration on muscle activity during exercise., (Copyright © 2022. Published by Elsevier Inc.)

Horses' muscular tension during acute stress remains unexplored. Our aim was to assess muscular, behavioral, cortisol, and hematocrit responses to social isolation (ISO), novel object exposure (NOV), and sham clipping (CLIP). Altered stress responses were expected. Eight mature Standardbred horses (four mares and four geldings) were exposed to acute stressors and a control period (CON) in a balanced, replicated 4×4 Latin Square experimental design with 3 min treatment periods and 10 min washout periods. Surface electromyography collected from the masseter, brachiocephalas, cervical trapezius, and longissimus dorsi was processed to derive average rectified value (ARV) and median frequency (MF) during the initial, middle, and final 30 s of treatments. ARV and MF data were log transformed then analyzed using a mixed model, repeated measures ANOVA along with plasma cortisol and hematocrit. Behavior data were analyzed using a negative binomial distribution mixed model ANOVA. CLIP resulted in greater (p < 0.05) log ARV in the masseter (1.5 + 1.5%, mean + SD) and brachiocepahlas (2.2 + 2.0%) than CON (-1.2 + 1.4%, 0.1 + 1.5%). ISO resulted in greater (p < 0.05) log ARV in the masseter (0.2 + 1.3%) and cervical trapezius (0.6 + 1.3%) than CON (-1.2 + 1.4%, -1.0 + 1.7%). ISO increased (p < 0.05) the total number of stress-related behaviors and hematocrit. No changes in cortisol were observed. We suggest that muscular tension can be used as an indicator of acute stress in horses. Incorporating muscle activity into an array of measurements may provide a more nuanced understanding of stress responses., (© 2022 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

Little published information exists on the horses in equine-assisted services (EAS), particularly their selection, longevity, and retirement. The purpose of this study was to characterize horses and procedures used in EAS. A pilot survey was developed using focus group discussions and distributed to Professional Association of Therapeutic Horsemanship, International (PATH Intl) centers in Florida ( n = 45, Part I) before further modification and distribution to members of PATH Intl., American Hippotherapy Association (AHA), eagala, and Certified Horsemanship Association (CHA) ( n = 26,000, Part II). Response rates were 36% (Part I) and 0.7% (Part II). Centers report a median of 10 (Part I) or 9 (Part II) horses and ponies. Selection procedures included initial screening (Part I = 100%, Part II = 96%), pre-purchase or pre-donation exam (I = 64%, II = 60%), acclimation period (I = 100%, II = 84%), trial period (I = 91%, II = 90%), and other (II = 11%). Horses remained active in programs for less than a year to over 20 years with the greatest number working 7-10 (Part I) or 1-6 (Part II) yr. In Part I of the study, behavior (44%) was the leading cause of retirement followed by unsoundness (33%). In Part II, unsoundness was the highest ranked response followed by behavior. Behavior, soundness, and health emerged as key factors in horse selection and retirement. Future work should focus on investigating these issues at an individual horse level.

metabolomics is the high-throughput, multiparametric identification and classification of hundreds of low molecular weight metabolites in a biological sample. Ultimately, metabolites are the downstream readouts of cellular signalling, transcriptomic and proteomic changes that can provide a comprehensive view of tissue and organismal phenotype. The popularity of metabolomics in human sport and exercise has been gaining over the past decade and has provided important insights into the energetic demands and mechanistic underpinnings of exercise and training. To the contrary, metabolomics in the field of equine exercise physiology is lagging despite the horse's superior aerobic and muscular capabilities, as well as its prominence in competitive sport. As such, this narrative review aims to describe metabolomics, its routine implementation, the various analytical methods applied and the state of its use in the equine athlete. Sufficient attention will be paid to methodological considerations, as well as gaps in the equine literature, particularly with regard to the skeletal muscle metabolome. Finally, there will be a brief discussion of the future directions and barriers to metabolomics use in the athletic horse. A thorough understanding of the metabolomics changes that occur in the equine athlete with exercise will undoubtedly help to improve horse management and health across the lifespan., (© 2020 Blackwell Verlag GmbH.)

The athletic horse, despite being over 50% muscle mass, remains understudied with regard to the effects of exercise and training on skeletal muscle metabolism. To begin to address this knowledge gap, we employed an untargeted metabolomics approach to characterize the exercise-induced and fitness-related changes in the skeletal muscle of eight unconditioned Standardbred horses (four male, four female) before and after a 12-week training period. Before training, unconditioned horses showed a high degree of individual variation in the skeletal muscle metabolome, resulting in very few differences basally and at 3 and 24 h after acute fatiguing exercise. Training did not alter body composition but did improve maximal aerobic and running capacities ( p < 0.05), and significantly altered the skeletal muscle metabolome ( p < 0.05, q < 0.1). While sex independently influenced body composition and distance run following training ( p < 0.05), sex did not affect the skeletal muscle metabolome. Exercise-induced metabolomic alterations ( p < 0.05, q < 0.1) largely centered on the branched-chain amino acids (BCAA), xenobiotics, and a variety of lipid and nucleotide-related metabolites, particularly in the conditioned state. Further, training increased ( p < 0.05, q < 0.1) the relative abundance of almost every identified lipid species, and this was accompanied by increased plasma BCAAs ( p < 0.0005), phenylalanine ( p = 0.01), and tyrosine ( p < 0.02). Acute exercise in the conditioned state decreased ( p < 0.05, q < 0.1) the relative abundance of almost all lipid-related species in skeletal muscle by 24 h post-exercise, whereas plasma amino acids remained unaltered. These changes occurred alongside increased muscle gene expression ( p < 0.05) related to lipid uptake ( Cd36 ) and lipid ( Cpt1b ) and BCAA ( Bckdk ) utilization. This work suggests that metabolites related to amino acid, lipid, nucleotide and xenobiotic metabolism play pivotal roles in the response of equine skeletal muscle to vigorous exercise and training. Use of these and future data sets could be used to track the impact of training and fitness on equine health and may lead to novel predictors and/or diagnostic biomarkers., (Copyright © 2020 Klein, McKeever, Mirek and Anthony.)

Sturkie's Avian Physiology is a highly regarded textbook for the study of comparative poultry physiology. Less well known, however, is the contribution of Paul D. Sturkie (1909-2002) as a pioneer in the experimental physiology of avian species. His seminal research on the cardiovascular and hemodynamic controls of chickens and egg-laying hens had a notable impact on the poultry industry and breeding practices of farmers. The purpose of this article is to highlight the contributions and practical insights of Paul D. Sturkie to the field of poultry science.

With the increase in the number of horses being used in Equine-Assisted Activities and Therapies (EAAT) programs and with the increasing concern for animal welfare, it is important to understand the impact of such interventions on the stress level and quality of life for the horses involved. The purpose of the present pilot study was to test the hypothesis that participation in EAAT would acutely alter physiological markers of stress and well-being, including plasma cortisol, plasma oxytocin, and heart rate variability (HRV), in horses and that symptoms of posttraumatic stress disorder (PTSD) would be reduced after five sessions of EAAT in veterans who had previously been diagnosed with PTSD. Nine healthy geldings, of various breeds, ages 10-23 years, conditioned and experienced as therapeutic riding horses, were selected to participate in the study. Of these, seven were selected at random to wear electrocardiogram units, and all nine were used for blood sampling to measure plasma cortisol and oxytocin. Each horse was randomly assigned to partner with a veteran for five EAAT sessions, 1 hour in duration. A standing control was conducted on a later date on which horses did not participate in EAAT. Measurement after 5 days of EAAT was conducted immediately after the end of the last session on day 5 using the Brief Symptom Inventory and the PCL-5 (a 20 item self-report measure of the Diagnostic and Statistical Manual of Mental Disorders - 5 for symptoms of PTSD). Two way repeated measure analysis of variance showed no significant day by time interactions for plasma cortisol (P = .821) or oxytocin (P = .861). There was a significant day by time interaction (P = .006) for heart rate (HR); where on day 1, HR (bpm) was significantly lower during the interaction with the veterans. There were no significant differences in HRV variables. Posttherapy measures in PTSD symptoms in veterans were significantly reduced except for interpersonal sensitivity (P = .08) and phobic anxiety (P = .17). There was an effect of EAAT on HR which was significantly reduced on day 2 during the actual EAAT session. Equine-Assisted Activities and Therapies had no effect on respiration rate and systolic or diastolic blood pressure in veterans involved in five sessions of EAAT, lasting 60 minutes in duration over the course of 5 days. Stress levels, as demonstrated by plasma cortisol concentrations and HRV, did not change in horses involved in EAAT sessions with veterans who had been previously diagnosed with PTSD. Furthermore, the horses used in this study did not demonstrate increased levels of well-being as demonstrated by the lack of change in plasma oxytocin concentrations after EAAT sessions. Symptoms of PTSD did change significantly in the veterans who participated in this study., (Copyright © 2018 Rutgers_The State University of New Jersey. Published by Elsevier Inc. All rights reserved.)

There are many choices for methods of extracting bacterial DNA for Next Generation Sequencing (NGS) from fecal samples. Here, we compare our modifications of a phenol/chloroform extraction method plus an inhibitor removal solution (C3) (ph/Chl+C3) to the PowerFecal® DNA Isolation Kit (MoBio-K). DNA quality and quantity coupled to NGS results were used to assess differences in relative abundance, Shannon diversity index, unique species, and principle coordinate analysis (PCoA) between biological replicates. Six replicate samples, taken from a single ball of horse feces manually collected from the rectum, were subjected to each extraction method. The Ph/Chl+C3 method produced 100× higher DNA yields with less shearing than the MoBio-K method. To assess the methods, the two method samples were sent for sequencing of the bacterial V3-V4 region of 16S rRNA gene using the Illumina MiSeq platform. The relative abundance of Bacteroidetes was greater and there were more unique species assigned to this group in MoBio-K than in Ph/Chl+C3 (P<0.05). In contrast, Firmicutes had greater relative abundance and more unique species in Ph/Chl+C3 extracts than in MoBio-K (P<0.05). The other major bacterial phyla were equally abundant in samples using both extraction methods. Alpha diversity and Shannon Weaver indices showed greater evenness of bacterial distribution in Ph/Chl+C3 compared with MoBio-K (P<0.05), but there was no difference in the OTU richness. Principle coordinate analysis (PCoA) indicated a distinct separation between the two methods (P<0.05) and tighter clustering (less variability) in Ph/Chl+C3 than in MoBio-K. These results suggest that the Ph/Chl+C3 may be preferred for research to identify specific Firmicutes taxa such as Clostridium, and Bacillus. However; MoBio-K may be a better choice for projects focusing on Bacteroidetes abundance. The Ph/Chl+C3 method required less time, but has some safety concerns associated with exposure and disposal of phenol and chloroform. While the MoBio-K may be better choice for researchers with less access to safety equipment like a fume hood., (Copyright © 2016 Elsevier B.V. All rights reserved.)

An increasing percentage of the equine population is more than 15 years old, many performing various athletic activities into their 20s. Studies of aged humans have led to a fine tuning of exercise prescription to promote fitness while preventing adverse and potentially dangerous effects of excessive exercise. However, limited data exist regarding the exercise capacity of aged horses. This article presents an overview of published studies on aging-induced decreases in physiologic function and exercise capacity in the horse. The information presented can be used as a guide for exercise prescription for the growing population of active older equine athletes., (Copyright © 2016 Elsevier Inc. All rights reserved.)

To test the hypotheses that erythropoietin (rhuEPO) treatment increases systemic hematocrit, maximal O2 uptake (VO2max, by elevated perfusive and diffusive O2 conductances) and performance five female horses (4-13 years) received 15 IU/kg rhuEPO (erythropoietin) three times per week for three weeks. These horses had been splenectomized over 1 year previously to avoid confounding effects from the mobilization of splenic red blood cell reserves. Each horse performed three maximal exercise tests (one per month) on an inclined (4°) treadmill to the limit of tolerance; two control trials and one following EPO treatment. Measurements of hemoglobin concentration ([Hb] and hematocrit), plasma and blood volume, VO2, cardiac output as well as arterial and mixed venous blood gases were made at rest and during maximal exercise. EPO increased resting [Hb] by 18% from 13.3 ± 0.6 to 15.7 ± 0.8 g/dL (mean ± SD) corresponding to an increased hematocrit from 36 ± 2 to 46 ± 2% concurrent with 23 and 10% reductions in plasma and blood volume, respectively (all P<0.05). EPO elevated VO2max by 20% from 25.7 ± 1.7 to 30.9 ± 3.4 L/min (P<0.05) via a 17% increase in arterial O2 content and 18% greater arteriovenous O2 difference in the face of an unchanged cardiac output. To achieve the greater VO2max after EPO, diffusive O2 conductance increased ∼ 30% (from 580 ± 76 to 752 ± 166 mL O2/mmHg/min, P<0.05) which was substantially greater than the elevation of perfusive O2 conductance. These effects of EPO were associated with an increased exercise performance (total running time: control, 216 ± 72; EPO, 264 ± 48 s, P<0.05). We conclude that EPO substantially increases VO2max and performance in the splenectomized horse via improved perfusive and diffusive O2 transport., (Copyright © 2016 Elsevier B.V. All rights reserved.)

This study tested the hypotheses that age-induced alteration in cortisol, ACTH, and glucose concentrations are due to differences in the response of the hypothalamic-pituitary-adrenal axis and that exercise training would attenuate these differences. Six old (22.0±0.7 yr; mean±SE) and 6 young (7.3±0.6 yr) unfit Standardbred mares ran 3 graded exercise tests (GXT): before (GXT1), after 8 wk of training (GXT2), and at study end at 15 wk (GXT3). Mares trained 3 d/wk at 60% maximum heart rate. Each mare underwent 5 endocrine stimulation tests pre- and posttraining: 1) control (CON), 2) adrenocorticotropin hormone (ACTHtest), 3) combined dexamethasone suppression/ACTH (DEX/ACTH), 4) dexamethasone suppression (DEX), and 5) combined DEX/corticotropin releasing factor (DEX/CRF). For CON, there was no difference in plasma cortisol between age groups pretraining (P=0.19), but young mares had a 102% higher mean (P=0.02) plasma cortisol concentration than old mares posttraining. The pretraining ACTHtest showed young mares had a 72% higher (P=0.05) overall plasma cortisol concentration compared to old. There was no overall age difference in cortisol in the posttraining ACTHtest, but old mares still had lower cortisol concentrations at 30 min during the test, suggesting decreased adrenal response to ACTH stimulation. There was no difference in cortisol response between old and young mares in DEX, DEX/ACTH, or DEX/CRF tests. Young mares had higher (P=0.02) overall plasma cortisol concentration posttraining in response to DEX/ACTH, but old mares showed no change. In CON and DEX/CRF, there were no age differences in plasma ACTH concentration, pre- or posttraining. Pretraining, there was no age difference in glucose response to DEX, but posttraining old mares had a 4% (P=0.04) lower overall plasma glucose concentration compared to young. Posttraining, old mares had lower mean plasma glucose concentrations during DEX compared to pretraining (P=0.02), but there was no change pre- vs. posttraining in young mares (P=0.19). Old and young mares had lower plasma glucose concentrations posttraining during DEX/ACTH (P<0.001 and P=0.05, respectively) and DEX/CRF (P<0.001 and P=0.003, respectively) compared to pretraining. Both the pituitary and adrenal glands experience a decline in function with age although the exact mechanisms behind such changes remain unknown. Exercise training facilitates the counteraction of these deficits.

Horses spend most of their day eating, standing, and occasionally exercising. Exercise can range from running in a pasture to athletic training. Under resting conditions, horses easily maintain the internal environment. The performance of work or exercise is a major physiologic challenge, a disturbance to homeostasis that invokes an integrative response from multiple organ systems. The response to exercise involves endocrine and neuroendocrine signaling associated with the short-term and adaptive control of many systems. The coordinated control of multiple physiologic variables is essential for achieving regulation to maintain the integrity of the internal environment of the body., (Copyright © 2011 Elsevier Inc. All rights reserved.)

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., (© 2010 EVJ Ltd.)

Reasons for Performing Study: Mangalarga-Marchador is a popular 4-gaited Brazilian horse breed; however, there is little information about their metabolic and physiological response to exercise., Objectives: To measure physiological and metabolic responses of the Mangalarga-Marchador to a simulated marcha field test and to compare these responses between 2 types of marcha gaits (picada and batida)., Methods: Thirteen horses were used in the study and randomly assigned to either the picada or batida gait for the simulated marcha field test (speed ∼ 3.2 m/s; 30 min; load ∼ 80 kg)., Measurements: Included body composition, heart rate (HR), respiratory rate (RR), glucose (GLUC), lactate (LACT), packed cell volume (PCV), total plasma protein (TPP), albumin, urea, creatinine, total and HDL cholesterol, triglycerides, creatine kinase, alanine, glutamate and glutamine (GLN). Measurements were obtained pretest (control/fasting), immediately after simulation (T(0)), and 15 (T(15)), 30 (T(30)) and 240 (T(240)) min after the simulation. Lactate (LACT) was measured at T(0), T(15) and T(30). Data were analysed using ANOVA, Tukey's test and t tests with significance set at P < 0.05., Results: Significant acute changes were observed in HR, RR, [GLUC], [LACT], [TPP], PCV and [GLN] (P<0.05) relative to control. Heart rate fell below 60 beats/min at T(15) and RR recovered to pretest values by T(240). Significant increases in [GLUC], [LACT], PCV and [TPP] and a decrease in [GLN] were observed at T(0). Treatment and interaction effects were also observed between marcha types and time of sampling for HR, RF, PCV, and [LACT] (P < 0.05). These parameters were large in picada., Conclusion: The simulation of field-test produced changes in some physiological and blood parameters in marcha horses, with some degree of dehydration during recovery period. Also, it was demonstrated that picada horses spend more energy when compared with batida horses at the the same speed., Potential Relevance: Batida horses spend less energy when compared with picada horses, which will need special attention in their training and nutritional management., (© 2010 EVJ Ltd.)

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., (© 2010 EVJ Ltd.)

Background: Muscle 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., Methods: College-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., Results: Compared 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., Conclusions: Consumption 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 purpose of this study was to test the hypothesis that sucralfate, a gastric ulcer medication, would alter plasma concentrations of total carbon dioxide (tCO2), lactate (LA), sodium (Na+), potassium (K+), chloride (Cl-) and total protein (TP), as well as calculated plasma strong ion difference (SID) and packed cell volume (PCV) in horses subjected to a simulated race test (SRT). Six unfit Standardbred mares (approximately 520 kg, 9-18 years) were used in a randomized crossover design with the investigators blinded to the treatment given. The horses were assigned to either a control (40-50 mL apple sauce administered orally (PO)) or a sucralfate (20 mg/kg bodyweight dissolved in 40-50 mL apple sauce administered PO) group. Each horse completed a series of SRTs during which blood samples were taken via jugular venipuncture at five sampling intervals (prior to receiving treatment, prior to SRT, immediately following exercise, and at 60 and 90 min post-SRT). During the SRTs, each horse ran on a treadmill fixed on a 6% grade for 2 min at a warm-up speed (4 m/s) and then for 2 min at a velocity predetermined to produce VO2max. Each horse then walked at 4 m/s for 2 min to complete the SRT. Plasma tCO2, electrolytes, LA, and blood PCV and TP were analysed at all intervals. No differences (P>0.05) were detected between control and sucralfate for any of the measured variables. There were differences (P<0.05) in tCO2, SID, PCV, TP, LA and electrolyte concentrations relative to sampling time. However, these differences were attributable to the physiological pressures associated with acute exercise and were not an effect of the medication. It was concluded that sucralfate did not alter plasma tCO2 concentration in this study., (Copyright (c) 2008 Elsevier Ltd. All rights reserved.)

Clenbuterol is a beta(2)-agonist and potent selective bronchodilator that is used to treat bronchospasm in the horse. The drug is normally administered to horses orally as a syrup formulation. Once absorbed into the systemic circulation, clenbuterol has the potential to cause many side effects, including a repartitioning effect and major alterations in cardiac and skeletal muscle function. Recent studies have also reported that clenbuterol can affect bone and the immune, endocrine and reproductive systems. A great deal of information has been published on the beneficial effects of short term therapeutic doses of clenbuterol on the equine respiratory system, although there is limited information about chronic administration, particularly since this has been associated with adverse physiological effects on other systems. This review summarizes the relevant understanding of clenbuterol for clinicians and horse owners who may administer this drug to pleasure and performance horses.

Glutamine is concentrated within skeletal muscle, where it has been proposed to play a regulatory role in maintaining protein homeostasis. The work presented here addressed the hypothesis that glutamine would be the most abundant free alpha-AA in plasma and skeletal muscle in the foal during the first year of life. Glycine, however, was the most abundant free alpha-AA in plasma at birth and between 3 and 12 mo of age. The concentration of glutamine, the second most abundant AA at birth, increased through the first 7 d (P < 0.05) and then returned to values similar to those at birth. This resulted in glutamine being the most abundant free alpha-AA in plasma from 1 d through 1 mo of age. The most abundant free alpha-AA in skeletal muscle at birth was glutamine, but the concentration fell by more than 50% by d 15 and continued to decrease, reaching about one-third of the original values by 1 yr of age (P < 0.05). Glutamine synthetase was barely detectable in skeletal muscle at birth, but the abundance increased rapidly within 15 d of birth. The concentration of glycine, the second most abundant alpha AA in muscle at birth, decreased by about 40% by d 15 (P < 0.05) and then stabilized at this value throughout the year. In contrast, glutamate, alanine, and serine concentrations, the third, fourth, and fifth most abundant free alpha-AA in muscle at birth, respectively, increased to new stable concentrations between 3 and 6 mo of age (P < 0.05). This resulted in alanine being the most abundant free alpha-AA in skeletal muscle at 12 mo of age, followed by glutamate, glutamine, and glycine. The decrease in intramuscular glutamine content, particularly during the first 2 wk after birth, is not compatible with a regulatory role for glutamine in muscle protein synthesis because it occurred at the time of maximum growth in these animals. The findings that, at certain times of development, glutamine was not the most abundant free alpha-AA in the foal is novel and signifies that intramuscular glutamine may have functions specific to muscle type and mammalian species.

In most mammalian species the developing fetus utilizes large amounts of glutamine derived both from the maternal circulation and synthesized de novo in the placenta. The present study was designed to determine the role of the placenta in glutamine synthesis in the horse. The placentae from eight Standardbred mares were sampled immediately after parturition together with additional tissues obtained at necropsy from three Standbred mares during diestrous. Glutamine synthetase protein was detectable in the non-pregnant horn of the placenta in amounts similar to those seen in gluteus muscle, but the amount in the pregnant horn was two times greater than in the non-pregnant horn. Glutamine was the second most abundant amino acid in amniotic fluid at a concentration of 310 +/- 26 micromole/L with that of glycine being 535 +/- 48 micromole/L. The most abundant amino acids in placental tissue were glycine (3,732 +/- 194 micromole/Kg), glutamate (3,500 +/- 343 micromole/Kg) and glutamine (2,836 +/- 208 micromole/Kg). The results illustrate the importance of glutamine to the equine fetus and establish that the placenta, particularly the pregnant horn, has considerable capacity for glutamine synthesis.

Glutamine is the most abundant free alpha-AA in the mammalian body, and large amounts of glutamine are extracted by both the fetus during pregnancy and the mammary gland during lactation. The work presented here addressed the hypothesis that there would be major changes in glutamine metabolism in the mare during the transition period, the time between late gestation, parturition, and early lactation. Eight foals were born to Standardbred mares provided with energy and protein at 10% above NRC recommendations, and foals remained with mares for 6 mo. During lactation, lean body mass decreased by 1.5% (P < 0.05), whereas fat mass was unchanged throughout gestation and lactation. There was a sharp increase in the concentration of most plasma metabolites and hormones after birth, which was due in part to hemoconcentration because of fluid shifts at parturition. Plasma glutamine concentration, however, was maintained at greater concentrations for up to 2 wk postpartum but then began to decrease, reaching a nadir at approximately 6 wk of lactation. Skeletal muscle glutamine content did not change, but glutamine synthetase expression was decreased at the end of lactation (P < 0.05). Free glutamine was highly abundant in milk early in lactation, but the concentration decreased by more than 50% after 3 mo of lactation and paralleled the decrease in plasma glutamine concentration. Thus, lactation represents a mild catabolic state for the mare in which decreased glutamine concentrations may compromise the availability of glutamine to other tissues such as the intestines and the immune system.

Glutamine plays important roles in the interorgan transport of nitrogen, carbon and energy but little is known about glutamine metabolism in the horse. In this study we determined the tissue distribution of glutamine synthetase expression in three Standardbred mares. Expression of glutamine synthetase was highest in kidney and mammary gland, and relatively high in liver and adipose tissue. Expression was lower in gluteus muscle, thymus, colon and lung, and much lower in small intestine, pancreas and uterus. The pattern of glutamine synthetase expression in the horse is similar to that of other herbivores and it is likely that skeletal muscle, liver, adipose tissue and lungs are the major sites of net glutamine synthesis in this species. Expression did not differ between adipose tissue depots but did vary between different muscles. Expression was highest in gluteus and semimembranous muscles and much lower in diaphragm and heart muscles. The concentration of intramuscular free glutamine was inversely correlated with expression of glutamine synthetase (r=-0.81, p=0.0017). The concentration of free glutamine was much higher in heart muscle (21.6+/-0.9 micromol/g wet wt) than in gluteus muscle (4.19+0.33 micromol/g wet wt), which may indicate novel functions and/or regulatory mechanisms for glutamine in the equine heart.

Eight mature (12 +/- 2 yr; MAT) and 5 older (22 +/- 2 yr; OLD) Standardbred mares were used to test the hypothesis that aging and exercise training would alter apoptosis in white blood cells and antioxidant status. The horses were housed indoors overnight (16 h/d) in 3 m x 3 m stalls and were turned out in a drylot during the day. They were fed a diet consisting of total mixed ration, hay cubes fed ad libitum or an equine senior diet plus grass hay. Horses were trained for 20 to 30 min/d, 3 to 5 d/wk for 8 wk at a submaximal work intensity between 60 to 70% of maximal heart rate. A graded exercise test (GXT; stepwise test until exhaustion) was performed before (GXT1) and after (GXT2) the 8 wk of training. During the GXT, blood samples and heart rate were taken at rest, 6 m/s, fatigue, and at 5 and 60 min postfatigue. Fatigue plasma lactate concentration was greater in MAT (19.3 +/- 1.5 at 10 m/s) compared with the OLD (10.9 +/- 1.2 mmol/L at 9 m/s; P = 0.008) horses. There was no effect of age or training on plasma lipid hydroperoxide (LPO) concentration. However, there was a positive correlation between LPO and plasma lactate concentration (r = 0.27, P = 0.006) during acute exercise. There was a greater concentration of total glutathione after GXT1 than after GXT2 (111.8 +/- 5.0 vs. 98.6 +/- 3.4 microM, respectively; P = 0.0002) for both age groups. Apoptosis was less (P = 0.002) in white blood cells of the MAT vs. the OLD group. These results demonstrate that older horses are under similar amounts of oxidative stress, measured by LPO, and have similar levels of glutathione in their systems compared with mature horses. The observation that more glutathione was needed during GXT1 for both groups of horses indicates that training helps horses adapt their system for the intense post-training exercise tests. The greater level of white blood cell apoptosis also indicates that older horses may be immune-compromised during exercise. However, research still needs to be performed regarding dietary supplementation in the aged horse.

One of the hallmarks of insulin resistance is a reduction in glucose transporter-4 (Glut-4) expression in adipose tissue but not in skeletal muscle. However, while Glut-4 has been demonstrated in skeletal and cardiac muscles in horses it has not been demonstrated in adipose tissue. The initial objectives of the present study were: (1) to test the hypothesis that Glut-4 expression would vary between selected key skeletal muscles; (2) to test the hypothesis that it would also vary between representative adipose tissue depots, and (3) to see whether expression would be greater in adipose tissue compared to muscle. Glut-4 expression was determined by Western blot using samples obtained from post mortem biopsies obtained from four muscles (gluteus medius, semitendinosus, heart, and diaphragm), and four adipose tissues (subcutaneous, retroperitoneal, mesenteric, and omental) in three horses. There were no differences (P>0.05) in Glut-4 protein expression between the muscles sampled. Likewise there were no differences (P>0.05) in Glut-4 protein expression between fat depots. There was a significant difference (P=0.03) when pooled means for Glut-4 expression in muscle (58.8+/-2.5 densitometry units) were compared with adipose tissue (115.8+/-15.7). This difference in Glut-4 expression in these two tissues with distinctly different metabolic reasons for taking up glucose may warrant further investigation to see if there are more pronounced differences in Glut-4 expression in muscle and adipose tissue in various populations of horses.

Six Standardbred (STB) mares (11+/-2 years, 521+/-77 kg; means+/-SD) performed an exercise trial (EX) where they underwent an incremental exercise test (GXT) as well as a parallel control trial (CON) to test the hypothesis that short-term, high intensity exercise would alter plasma concentrations of glucose, leptin, adiponectin, ghrelin, insulin and cortisol. Plasma samples were taken before (0 min), during (last 10s at 6, 8m/s, and the velocity eliciting VO(2max)), and after exercise (2, 10, 30, 60 min; 12 and 24h post-GXT). A second set of blood samples was collected before and after an afternoon meal given at 1515 h (at 1500, 1514, 1530, and 1545 h). Data were analyzed using ANOVA for repeated measures and Tukey's test. During the GXT, there were no changes (P>0.05) in the plasma concentrations of glucose, leptin, adiponectin or ghrelin. However, there was a 29% increase (P<0.05) in mean plasma cortisol concentration and a 35% decrease (P<0.05) in mean plasma insulin concentration. Substantial increases (P<0.05) in the mean plasma concentrations of glucose and cortisol of 36% and 102%, respectively, were seen in the EX trial during the first 60 min post-GXT. Plasma leptin concentration, measured at the 24h post-GXT time point, was 20% lower (P<0.05) during the EX trial compared with the parallel time point in the standing control (CON) trial. Plasma ghrelin concentration was 37% lower (P<0.05) in the EX trial compared with CON before and after the afternoon meal, but was 43% higher (P<0.05) 12h post-GXT. There were no differences between EX and CON for plasma concentrations of insulin or adiponectin during recovery. It was concluded that short-term high intensity exercise alters plasma leptin and ghrelin concentrations in STB mares post-exercise, which may signal the exercised animals to alter energy intake.

Concentrations of hormones related to energy homeostasis may differ between populations with varied body compositions, acting as signals to increase or decrease energy intake and/or expenditure. How these parameters correlate with body composition in horses and how they vary in fit (F) versus unfit (UF) Standardbred racehorses is unclear. The purpose of this study was to test the hypothesis that plasma concentrations of glucose (GLU), insulin (INS), cortisol (CORT), ghrelin (GHRL), adiponectin (ADIP) and leptin (LEP) would be correlated with body composition and differ in fit (F) versus unfit (UF) horses. Fasting plasma samples were taken from 12 unfit (11 +/- 2 years, 521 +/- 77 kg; mean +/- SD) and 34 fit (4 +/- 2 years, 475 +/- 83 kg) Standardbred horses. GHRL, LEP, ADIP, INS and CORT concentrations were measured using radioimmunoassay. GLU concentration was measured using colorometric kits. Body composition data included body weight, body condition score (BCS), and percent fat (%fat) calculated using rump fat thickness measured ultrasonically and the Westervelt equation. Data were analyzed using Pearson Product moment and Student's t tests. There were no differences (P>0.05) between F and UF horses for the plasma concentrations of CORT (69 +/- 14 versus 76 +/- 23 microg/dL), INS (7.2 +/- 3.5 versus 7.1 +/- 1.8 microIU/mL) or GLU (90 +/- 6 versus 86 +/- 7 mg/dL). Plasma GHRL and ADIP concentrations were greater (P<0.05) in F versus UF horses (54 +/- 27 versus 33 +/- 17 pg/mL and 1820 +/- 276 versus 1333 +/- 249 ng/mL, respectively), while plasma LEP was lower in F versus UF (1.0 +/- 0.6 versus 4.4 +/- 2.4 ng/mL, P<0.001). BCS and %fat were lower in F versus UF horses (4.8 +/- 0.3 versus 6.7 +/- 0.5 and 11.9 +/- 1.6 versus 15.4 +/- 2.5%, respectively), with no correlation between %fat and GHRL (-0.12, P>0.05), although there was a positive correlation between %fat and LEP (+0.72, P<0.05), and a negative correlation between %fat and ADIP (-0.40, P<0.05). The data show that in comparing fit and unfit horses, there are variations in body composition as well as concurrent and substantial differences in the concentrations of hormones, cytokines, and other parameters related to the control of appetite and feed intake.

Plasma concentrations of adiponectin and leptin were measured in 23 mature Standardbred mares (age: 10+/-3 years) and 12 weanling fillies (10 Quarter Horse/Belgian crossbreds and two Quarter Horses; aged: 4+/-3 months) to test the hypothesis that adipocytokines are proportional to adiposity in horses. Rump fat thickness was measured using B-mode ultrasound and percent body fat (% fat) calculated using a published formula for the estimation of fatness in horses. Plasma adiponectin and leptin were determined using radioimmunoassay. In the absence of purified equine adiponectin or leptin, results were expressed as human equivalents (HE) of immunoreactive (ir) adipocytokines. Plasma ir-leptin HE concentration was positively correlated (r=0.543; P<0.001) with percent body fat and (r=0.556; P<0.001) to fat mass in all horses. The plasma ir-leptin HE concentration was lower (P=0.03) in weanlings (1.90+/-0.34 ng/mL HE) than in mature mares (3.47+/-0.50 ng/mL HE). The ratio of ir-adiponectin HE to ir-leptin HE was negatively correlated (r=-0.621; P<0.001) to percent fat and (r=-631; P<0.001) to fat mass in all horses. The ratio of ir-adiponectin HE to ir-leptin HE was higher (P<0.001) in weanlings (3.04+/-0.51) than mature mares (1.03+/-0.13). These data suggest that leptin is proportional while adiponectin is inversely proportional to adiposity in horses.

Objective: The efficacy of KAATSU training has been demonstrated in human athletes, both as a therapeutic method as well as a training aid. The purpose of this study was to investigate the effects of slow walk training combined with restriction of muscle blood flow (KAATSU) on muscle and tendon size., Methods: Six healthy, unfit Standardbred mares performed walking (240 m/min for 10 min and then 5 min recovery) with KAATSU, and 6 mares performed walking without KAATSU. A specially designed elastic cuff1 was placed at the most proximal position of the forelegs and inflated to a pressure of 200-230 mmHg throughout the walking and recovery sessions. The training was conducted once a day, 6 days/week for 2 weeks. Skeletal muscle thickness and tendon thickness were measured using B-mode ultrasound at baseline and after 2 weeks of training. Venous blood samples were obtained before the first acute exercise and 5, 15 and 60 min afterwards. Serum somatotropin concentration was determined using a commercially available equine-specific ELISA kit., Results: The acute increase in plasma somatotropin was 40% greater (P<0.05) in the KAATSU-walk group than in the Control-walk group 5 min after exercise and remained elevated (P<0.05) at 15 and 60 min post exercise compared with the Control-walk group. After 2 weeks of training, muscle thickness increased (P<0.05) 3.5% in the KAATSU-walk group but did not change in the Control-walk group (0.7%). Tendon thickness did not change (P>0.05) in either group., Conclusions: These data demonstrate that KAATSU training can induce muscle hypertrophy in horses and suggest that KAATSU training may provide significant therapeutic/ rehabilitative value in horses, as has been shown in man.

Reasons for Performing Study: A large percentage of performance horses develop gastric ulcers and many of those horses are treated with omeprazole. Unfortunately, no data have been published on the effects of the drug on markers of performance in animals without ulcers., Hypothesis: Omeprazole would alter markers of aerobic and anaerobic performance., Methods: Ten unfit, healthy, ulcer free, Standardbred mares were administered either control (CON; oral apple sauce, 20 ml) or omeprazole (OP; oral paste, 4 mg/kg bwt s.i.d.) in a random crossover fashion with the investigators blind to the treatment. Treatments were administered for 7 days prior to performing an incremental exercise test (GXT) on a high-speed treadmill. Endoscopic examinations were performed just prior to the trial to verify that the mares were ulcer-free. During the GXT, the mares ran on a treadmill up a 6% grade to measure maximal oxygen consumption (VO2max), run time (RT), velocity at VO2max, maximal velocity (Vmax), packed cell volume (PCV), plasma lactate concentration (LA) and plasma protein concentration (TP). Measurements were recorded at rest, at the end of each 1 min step of the GXT and at 2 and 5 min post GXT. Data were analysed using ANOVA for repeated measures and t tests for paired comparisons., Results: There was no effect (P>0.05) of omeprazole on VO2max; velocity at VO2max; RT; Vmax; 2 min recovery plasma LA. Nor were there any changes (P>0.05) in the relationship between treadmill speed and VO2, PCV, TP, or plasma LA., Conclusions: Omeprazole does not appear to improve physiological markers of performance in healthy, ulcer free horses., Potential Relevance: These data may benefit various authorities responsible for deciding administration and timing policies of omeprazole as well as clinicians and horse owners.

Reasons for Performing Study: Ageing appears to affect immune and neuroendocirne function in horses and response to acute exercise. No studies have examined the combined effects of training and ageing on immune and neuroendocirne function in horses., Hypothesis: To ascertain whether training and age would affect the plasma beta-endorphin (BE) and cortisol (C) as well as immune function responses to acute exercise in Standardbred mares., Methods: Graded exercise tests (GXT) and simulated race tests (SRT) were performed before and after 12 weeks training at 60 % HRmax. BE and C were measured at rest and at 5, 10, 20, 40, 60 and 120 min post GXT. Leucocyte cell number, CD4+ and CD8+ lymphocyte subsets, and mitogen stimulated lymphoproliferative response (LPR), were measured in jugular blood before and after the SRTs., Results: Cortisol rose by 5 min post GXT in young (Y) and middle-age (MA) mares (P<0.05) and remained elevated until 40 and 60 min post GXT, respectively during both pre- and post training GXT. There was no rise in C in old (0) mares after either GXT (P>0.05). Pretraining BE rose (P<0.05) by 5 min post GXT in all mares. After training, BE was higher in Y and O vs. MA (P<0.05) at 5 min post GXT. Post training BE was higher at 5 min post GXT in Y and O vs. pretraining (P<0.05). After SRT, lymphocyte number rose in all mares (P<0.05); however, lower lymphocyte numbers (P<0.05) were seen in MA vs. Y and O vs. MA (P<0.05). The O had reduced LPR to Con A and PHA stimulation (P<0.05) compared to Y and MA after the SRT after both pre- and post training SRT. LPR to PWM was lower (P<0.05) in O vs. Y and MA after the pretraining SRT. Training caused an increase in resting LPR to PWM in MA only (P<0.05)., Conclusion: Both age and training altered the plasma beta-endorphin and cortisol responses as well as and immune responses to acute exercise., Potential Relevance: This study provides important information on the effects of ageing and training that will aid in the management and care of an increasing number of active older horses.

Reasons for Performing Study: Heart rate is one of the most commonly measured variables in equine exercise physiology and relative exercise intensity commonly expressed as % of maximal heart rate. A number of influences affect maximal heart rate (HRmax), including age of the horse but other factors have not been described., Objectives: To determine if fitness, health status, gender, breed, athletic use, body mass, in addition to age, are predictive of HRmax in the horse., Methods: Maximal heart rate data from 328 horses which underwent treadmill exercise tests at 5 different laboratories were obtained retrospectively. Univariable linear regression analyses were performed on individual variables. Multiple linear regression analysis using a backward elimination modelling procedure was then used to relate the observed HRmax values simultaneously with different predictive variables. Variables were retained in the final regression model if they or any of their categories were significantly predictive of HRmax at P<0.05 and if there was a significant collective contribution to the model from inclusion of each variable, also at P<0.05., Results: Age, fitness status, laboratory, gender and breed/use (combined category) were all statistically significantly predictive of HRmax. Together these variables accounted for 41% of the variance in HRmax. Age alone accounted for only approximately 13% of the variation between horses in HRmax. Neither body mass nor health status were significantly predictive., Conclusions: HRmax in the horse declines with age but is also influenced by other factors. As the factors investigated accounted for only 41% of the variation between horses, other unidentified variables with a strong influence on HRmax remain to be identified., Potential Relevance: Factors such as fitness, age, gender, breed and use need to be considered when interpreting estimates or measurements of HRmax.

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.

Reasons for Performing Study: Horses in training tend to become inappetant; however, the mechanism responsible for this training-induced inappetance is not known., Hypothesis: Training and/or ulcers alter the feed intake (FI) and hormonal and/or biochemical (active ghrelin, leptin, glucose, insulin and cortisol) responses to acute high intensity exercise., Methods: Eight Standardbred mares underwent 3 interval exercise tests (IET) and 3 parallel control tests (CON) before (IET1) and after 8 weeks of training (IET2) and after treatment for gastric ulcers (IET3). Plasma samples were taken before (0 min), during (last 10 sec of velocities eliciting 40, 100 and 20% VO2max), and after (30 min, 60 min, 24 h) exercise (EX) or CON tests for RIA and colorimetric measurement of the concentrations of the above parameters. Samples were also collected before and after feeding. Horses were trained at a work intensity of 70% HRmax for 30 min/day, 5 days per week with FI measured daily., Results: There were no changes (P>0.05) in any variable during the parallel control trials. However, there was a mismatch between FI and digestible energy (DE) requirements (P<0.05) with EX horses not meeting their DE requirements during the post training IETs. During all IETs, ghrelin, glucose and cortisol increased (P<0.05) during EX. Leptin only increased (P<0.05) during EX in the post training IETs. Insulin remained low during EX, but increased (P<0.05) post EX., Conclusion: High intensity exercise appeared to be associated with decreases in FI and alterations of leptin and ghrelin., Potential Relevance: More research is needed to determine if there is a relationship between alterations of these hormones and changes in FI in horses that lose weight while in training.

This study tested the hypothesis that grain and intravenous dextrose challenges would alter plasma concentrations of active ghrelin, adiponectin, leptin, glucose, insulin, and cortisol in Standardbred mares. To deliver 0.5 g of glucose (dextrose solution for the intravenous test)/kg of BW, mares received intravenous dextrose (50% solution) or oral grain administration in 2 trials. In response to the oral grain challenge, plasma glucose and insulin concentrations increased (P < 0.001) by 56 and 802%, respectively. Plasma ghrelin concentration initially decreased (P < 0.001) by 40%, then subsequently increased (P < 0.05) from its nadir by 259%. Plasma leptin concentration decreased (P = 0.002) 17% compared with baseline. There was no change (P = 0.34) in plasma adiponectin concentration in response to oral grain challenge; however, plasma cortisol concentrations decreased (P < 0.001) by 24%. In response to the intravenous dextrose challenge, plasma glucose and insulin concentrations increased (P < 0.001) by 432 and 395%, respectively. Plasma active ghrelin concentration initially decreased (P < 0.001) by 56%, then subsequently increased (P < 0.001) from its nadir by 314%. Plasma leptin concentration also increased (P < 0.001) by 33% compared with baseline. There was no change (P = 0.18) in plasma adiponectin concentration throughout the dextrose challenge. Plasma cortisol concentration increased (P = 0.027) by 20%. Hence, oral grain and intravenous nutrient challenges have the ability to alter variables potentially related to energy metabolism in mares, with acute changes in glucose and insulin possibly modulating changes in ghrelin and leptin.

Twelve Standardbred mares underwent blood sampling for 24 h to test the hypothesis that there is diurnal variation of humoral mediators of peripheral energy balance including active ghrelin, adiponectin, leptin, glucose, insulin, and cortisol. The experiment was conducted under acclimated conditions. Grass hay and pelleted grain were provided at 0730 and 1530. Plasma concentrations of active ghrelin and leptin concentrations both peaked (47.3 +/- 6.5 pg/ mL and 5.9 +/- 1.1 ng/mL, respectively; P < 0.05) at 1550, 20 min after feeding. Active ghrelin decreased (P < 0.05) to 28.9 +/- 4.5 pg/mL overnight. The nadir of leptin (4.6 +/- 0.9 ng/mL) occurred at 0650. Neither hormone showed variation (P > 0.05) after the morning feeding. Plasma glucose and insulin concentrations increased (P < 0.05) in response to feeding; however, the morning responses (glucose = 96.9 +/- 2.6 mg/dL; insulin = 40.6 +/- 7.3 uIU/mL) were greater (P < 0.05) than the afternoon responses (glucose = 89.9 +/- 1.8 mg/dL; insulin = 23.2 +/- 4.3 uIU/mL at 180 and 60 min after feeding, respectively). Cortisol concentrations increased (P < 0.05) during the morning hours, but did not respond to feeding, whereas adiponectin concentrations remained stable throughout the study. Hence, active ghrelin and leptin may be entrained to meal feeding in horses, whereas adiponectin seems unaffected. We concluded that there seems to be a diurnal variation in glucose and insulin response to a meal in horses. Furthermore, elevated glucose and insulin concentrations resulting from the morning feeding may be responsible for the increase in leptin concentration in the afternoon.

Effects of longitudinal exercise training and acute intensive exercise (simulated race test) on immune function have not been reported in horses. Clenbuterol, a beta2-adrenergic agonist, is used to manage inflammatory airway disease in horses. This study investigated the interaction of 8 wk of exercise training with or without 12 wk of clenbuterol administration in horses. Twenty-three untrained standardbred mares (10 +/- 3 yr, Mean +/- SE) were used and divided into four experimental groups. Horses given clenbuterol plus exercise (CLENEX; n = 6) and clenbuterol alone (CLEN; n = 6) received 2.4 microg/kg BW of clenbuterol twice daily (in an average volume of 20 mL) on a schedule of 5 d on and 2 d off for 12 wk. The CLENEX group was also aerobically trained 3 d/wk. Mares given exercise alone (EX; n = 5) were aerobically trained for 3 d/wk, and the control group (CON; n = 6) remained sedentary. Both EX and CON horses were administered similar volumes (approximately 20 mL) of molasses twice daily. A simulated race test (SRT) resulted in an elevation in lymphocyte number postexercise (P < 0.05). There was no significant difference after acute exercise in either monocyte or granulocyte number. Acute exercise resulted in a decrease (P < 0.05) in the percentage of CD4+ and an increase (P < 0.05) in the percentage of CD8+ cells. The SRT resulted in a decreased lymphoproliferative response to pokeweed mitogen (P < 0.05). A SRT had no effect on antibody production in response to equine influenza vaccine. The EX group demonstrated greater cortisol concentrations at rest and at all other time points postexercise after completing the training regimen compared with CLENEX horses (P < 0.05). Preexercise (SRT) peripheral blood monocyte number was lower in CLENEX horses than in other treatment groups (P < 0.05). Clenbuterol and exercise training did not significantly affect post-SRT changes in leukocyte numbers. Exercise training resulted in a decrease (P < 0.05) in the percentage of CD8+ cells post-SRT compared with other groups, but the percentage of CD4+ cells was not altered by either clenbuterol or exercise conditioning. Lymphocyte proliferative response was not affected by clenbuterol or exercise treatment. Horses demonstrated responses to bouts of acute exercise as noted with other species, namely humans and rodents.

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.

The purpose of this study was to examine changes in myosin heavy chain (MHC) composition due to chronic clenbuterol administration with or without exercise in mares. Unfit Standardbred mares (aged 10+/-3 years) were divided into four groups: clenbuterol (2.4 micro/kg BW twice daily) plus exercise (3 days/week for 20 min at 50% VO(2max); CLENEX; n=6), clenbuterol only (CLEN; n=6), exercise only (EX; n=5), and control (CON; n=6). Muscle biopsies were obtained from gluteus medius muscle before and after the eight-week training/administration period. MHC composition was determined via SDS gel electrophoresis and quantified using a scanning and densometric system. CLENEX and CLEN exhibited significant (P<0.05) MHC changes while EX and CON did not. MHC type IIA decreased (29.8+/-6.1 to 19.3+/-4.0%, CLENEX; and 36.8+/-12.4 to 26.4+/-7.9%, CLEN) and MHC type IIX increased (59.4+/-7.2 to 71.8+/-5.8%, CLENEX; and 50.5+/-12.5 to 62.0+/-9.3%, CLEN). Chronic clenbuterol administration with and without exercise resulted in a significant shift in MHC profile in Standardbred mares.

Clenbuterol is a beta2-adrenoceptor agonist primarily used for treating bronchospasm and alleviating the symptoms of chronic obstructive pulmonary disease (COPD) in the horse. In other species (rats, mice, sheep, and cattle), chronic high doses of clenbuterol (typically in the milligram per kilogram body weight range) has been shown to cause a muscle directed protein anabolic response. Clenbuterol can also modify muscle fibre composition and therefore potentially affect muscle function. This has implications for the performance of exercising horses being treated with therapeutic doses of clenbuterol (typically in the microgram per kilogram body weight range) for bronchospasm or COPD. It is not known whether clenbuterol treatment affects muscle fibre function in horses. The purpose of this study was to examine the effects of a therapeutic dose of clenbuterol, with and without exercise, on the contractile activation characteristics of single membrane permeabilized fibres prepared from muscle biopsies. We tested the hypothesis that therapeutic treatment with clenbuterol would not affect muscle fibre function. Unfit Standardbred mares were treated for 8 weeks with; clenbuterol (2.4 microg/kg twice/day, 5 days/week) plus exercise (20 min at 50% VO2(max) 3 d/wk; CLENEX), clenbuterol only (CLEN), or exercise only (EX). Muscle biopsies were taken from the gluteus medius muscle before and after treatment and stored in a glycerol-based solution to prepare permeabilized muscle fibres. The force-pCa relationship for fibres from CLEN horses was steeper (P < 0.05) indicative of greater cooperative interactions within the thin filament, however, fibre sensitivity to Ca2+ was unchanged. In contrast, the steepness of the force-pCa relationship was not changed in fibres from EX and CLENEX horses and Ca2+ sensitivity was also unaffected. Rigor force, activation in the absence of ATP, was not affected by any treatment indicating an approximately equivalent number of participating cross-bridges during activation. The results indicate that a therapeutic dose of clenbuterol to Standardbred horses does not affect the Ca(2+)-activated contractile characteristics of isolated muscle fibres.