An optimally functional musculoskeletal system is crucial for athletic performance and even minor perturbations can limit athletic ability. The introduction of the muscle biopsy technique in the 1970s created a window of opportunity to examine the form and function of equine skeletal muscle. Muscle histochemical and biochemical analyses have allowed characterization of the properties of equine muscle fibres and their influence on, and adaptation to, physical exertion. Analyses of exercise responses during standardized treadmill exercise and field studies have illustrated the role of cellular energetics in determining athletic suitability for specific disciplines, mechanisms of fatigue, adaptations to training and the affect of diet on metabolic responses. This article provides a review of the tools available to study muscle energetics in the horse, discusses the muscular metabolic pathways and summarizes the energetics of exercise.qualitative assessment of the effects of exercise, training and dietary manipulation on substrate metabolism at the level of the muscle and the whole body. Muscle biopsy techniqueThe percutaneous needle muscle biopsy technique was introduced to equine research by Lindholm and Piehl 4 as well as by Snow and Guy 5 . Since that time, it has proved to be an invaluable tool in defining the histological, histochemical and biochemical properties of equine skeletal muscle. Standardization of the site of the muscle biopsy is imperative because equine skeletal muscles have a heterogeneous distribution of muscle fibre types within the muscle 6 . Deeper regions within locomotory muscles have contractile and metabolic characteristics similar to those of postural muscles 7-9 . In addition, fibre types vary among different muscles in the same horses as well as across horses and breeds [10][11][12][13][14][15][16][17] . The selection of the specific muscle to biopsy therefore is of critical importance and will depend on its propulsive or postural role. Many studies of equine athletes utilize the gluteus medius muscle or the semitendinosus muscle because of its importance in locomotion and demonstrated metabolic adaptations to exercise and training [18][19][20][21][22][23][24][25][26] . Other investigators have also used the triceps brachii, along with the masseter 26 (as a non-exercise muscle control sample site).When sampling site and depth are consistent and potentially involve several sites 21,23,24,27 , repeatable results are obtained 28 . Muscle biopsy has provided a wealth of information regarding the histochemical, biochemical and metabolic properties of various muscles within the same horse 9,29,30 , between horses 11 and breeds 15,21 as well as responses to exercise 25,31,32 . A detailed review of the technique for performing percutaneous needle muscle biopsy and sample preparation can be obtained elsewhere [3][4][5]21,22,33 .

Reasons for performing the study: In human exercise physiology, the current gold standard for evaluating aerobic capacity is the measurement of oxygen consumption (VO2) and maximal oxygen uptake (VO2max). The evaluation of VO2 in horses is performed in some laboratories equipped with a treadmill but has only been exceptionally reported in field conditions because of the lack of adapted equipment. Objectives: The aim of this study was (1) to assess the feasibility of VO2 measurement on the track using a recently validated portable breath-by-breath gas analyser system adapted to horses (Cosmed K4b2® and Equimask®), (2) to compare these results with those obtained during a treadmill exercise test and (3) to study correlations between VO2 and physiological parameters usually measured in field condition such as heart rate (HR), lactataemia (LA) and the speed at which HR equals 200 beats per minute (bpm) (V200) or LA 4 mmol l− 1 (VLA4). Methods: Five healthy Standardbred trotters in training were submitted to two stepwise incremental exercise tests, one driven on the racetrack and the other on a high-speed treadmill with a 4% incline. Speed (v), HR, ventilatory parameters and VO2 were continuously recorded throughout the duration of the tests and LA was evaluated after each step. Results: All horses completed the test satisfactorily after an initial acclimatization to the mask. There were marked individual differences in ventilatory strategy, and breathing frequency (Rf) at the higher levels of exercise was noticeably low. The VCO2 measurements were incoherent. There were no significant differences between track and treadmill maximal data obtained during the last step [VO2peak (track: 139.9 ± 8.9 ml kg− 1 min− 1; treadmill: 139.9 ± 13.4 ml kg− 1 min− 1), LAmax (track: 6.5 ± 1.6 mmol l− 1; treadmill: 7.3 ± 3.0 mmol l− 1), HRmax (track: 229 ± 6.2 bpm; treadmill: 222 ± 13 bpm)], although the maximal speed required to reach similar workloads was significantly higher on the track (11.9 ± 0.6 m s− 1vs. 9.7 ± 0.4 m s− 1). The correlation between VO2 and HR (r = 0.87; P < 0.001) and VO2 and LA (r = 0.75; P < 0.0001) during both tests was good but no correlation was found between VO2peak and HRmax, LAmax, V200 or VLA4. Conclusions: This is the first report of a practical portable system to measure VO2 and ventilation continuously during high-speed field exercise tests. However, current mask design markedly influences ventilation and could have prohibited the attainment of VO2max. Furthermore, consistent VCO2 measurements should be implemented by the manufacturers. Potential relevance: Continuous breath-by-breath ventilation and VO2 measurements can be recorded in horses in the field at submaximal levels. With necessary adaptations to the system entailed, this study opens new perspectives in the analysis of physiological and metabolic mechanisms of exercise in the equine species in genuine track conditions.

Acer pseudoplatanus is a worldwide-distributed tree which contains toxins, among them hypoglycin A (HGA). This toxin is known to be responsible for poisoning in various species, including humans, equids, Père David’s deer and two-humped camels. We hypothesized that any herbivore pasturing with A. pseudoplatanus in their vicinity may be at risk for HGA poisoning. To test this hypothesis, we surveyed the HGA exposure from A. pseudoplatanus in species not yet described as being at risk. Animals in zoological parks were the major focus, as they are at high probability to be exposed to A. pseudoplatanus in enclosures. We also searched for a toxic metabolite of HGA (i.e., methylenecyclopropylacetyl-carnitine; MCPA-carnitine) in blood and an alteration of the acylcarnitines profile in HGA-positive animals to document the potential risk of declaring clinical signs. We describe for the first instance cases of HGA poisoning in Bovidae. Two gnus (Connochaetes taurinus taurinus) exposed to A. pseudoplatanus in their enclosure presented severe clinical signs, serum HGA and MCPA-carnitine and a marked modification of the acylcarnitines profile. In this study, even though all herbivores were exposed to A. pseudoplatanus, proximal fermenters species seemed less susceptible to HGA poisoning. Therefore, a ruminal transformation of HGA is hypothesized. Additionally, we suggest a gradual alteration of the fatty acid metabolism in case of HGA poisoning and thus the existence of subclinical cases.

Background: Within the animal kingdom, horses are among the most powerful aerobic athletic mammals. Determination of muscle respiratory capacity and control improves our knowledge of mitochondrial physiology in horses and high aerobic performance in general.Methodology/Principal Findings: We applied high-resolution respirometry and multiple substrate-uncoupler-inhibitor titration protocols to study mitochondrial physiology in small (1.0-2.5 mg) permeabilized muscle fibres sampled from triceps brachii of healthy horses. Oxidative phosphorylation (OXPHOS) capacity (pmol O 2 Ns 21 Nmg 21 wet weight) with combined Complex I and II (CI+II) substrate supply (malate+glutamate+succinate) increased from 77618 in overweight horses to 103618, 122615, and 129612 in untrained, trained and competitive horses (N = 3, 8, 16, and 5, respectively). Similar to human muscle mitochondria, equine OXPHOS capacity was limited by the phosphorylation system to 0.8560.10 (N = 32) of electron transfer capacity, independent of fitness level. In 15 trained horses, OXPHOS capacity increased from 119612 to 134637 when pyruvate was included in the CI+II substrate cocktail. Relative to this maximum OXPHOS capacity, Complex I (CI)-linked OXPHOS capacities were only 50% with glutamate+malate, 64% with pyruvate+malate, and 68% with pyruvate+malate+glutamate, and ,78% with CII-linked succinate+rotenone. OXPHOS capacity with glutamate+malate increased with fitness relative to CI+II-supported ETS capacity from a flux control ratio of 0.38 to 0.40, 0.41 and 0.46 in overweight to competitive horses, whereas the CII/CI+II substrate control ratio remained constant at 0.70. Therefore, the apparent deficit of the CI-over CII-linked pathway capacity was reduced with physical fitness.Conclusions/Significance: The scope of mitochondrial density-dependent OXPHOS capacity and the density-independent (qualitative) increase of CI-linked respiratory capacity with increased fitness open up new perspectives of integrative and comparative mitochondrial respiratory physiology.

Introduction:The impact of eccentric exercise on mitochondrial function has only been poorly investigated and remains unclear. This study aimed to identify the changes in skeletal muscle mitochondrial respiration, specifically triggered by a single bout of eccentric treadmill exercise. Methods: Male adult mice were randomly divided into eccentric (ECC; downhill running), concentric (CON; uphill running), and unexercised control groups (n = 5/group). Running groups performed 18 bouts of 5 min at 20 cm•s −1 on an inclined treadmill (±15°to 20°). Mice were sacrificed 48 h after exercise for blood and quadriceps muscles collection. Deep proximal (red) and superficial distal (white) muscle portions were used for high-resolution respirometric measurements. Results: Plasma creatine kinase activity was significantly higher in the ECC compared with CON group, reflecting exercise-induced muscle damage (P < 0.01). The ECC exercise induced a significant decrease in oxidative phosphorylation capacity in both quadriceps femoris parts (P = 0.032 in proximal portion, P = 0.010 in distal portion) in comparison with the CON group. This observation was only made for the nicotinamide adenine dinucleotide (NADH) pathway using pyruvate + malate as substrates. When expressed as a flux control ratio, indicating a change related to mitochondrial quality rather than quantity, this change seemed more prominent in distal compared with proximal portion of quadriceps muscle. No significant difference between groups was found for the NADH pathway with glutamate or glutamate + malate as substrates, for the succinate pathway or for fatty acid oxidation. Conclusions: Our data suggest that ECC exercise specifically affects pyruvate mitochondrial transport and/or oxidation 48 h after exercise, and this alteration mainly concerns the distal white muscle portion. This study provides new perspectives to improve our understanding of the mitochondrial adaptation associated with ECC exercise.