Comparison of gluteus medius muscle activity in Haflinger and Noriker horses with polysaccharide storage myopathy

Type 1 polysaccharide storage myopathy caused by genetic mutation in the glycogen synthase 1 gene is present in many breeds including the Noriker and Haflinger horses. In humans, EMG has already been used to document changes in the muscle activity patterns of patients affected by human glycogen storage disorders. Therefore, the aim of the present study was to describe gluteus muscle activity with surface electromyography (sEMG) in Haflinger and Noriker horses with known GYS1 mutation status during walk and trot. Thirty-two horses (11 Haflinger and 21 Noriker horses) with homozygous non-affected (GG), heterozygous affected (GA) and homozygous affected (AA) status of GYS1 mutation without overt clinical signs of any myopathy were selected for the current study. Using surface electromyography gluteus medius muscle activity at walk and at trot was measured, and muscle activity was described in relation to the maximum observed value at the same sensor and the same gait. In order to further describe the signals in detail comprising both frequencies and amplitudes, the crossings through the baseline and the 25, 50 and 75 percentile lines were determined. The result of the relative muscle activity did not show a consistent difference between affected and non-affected horses. Genetically affected (GA and AA) horses showed significantly less density of muscle activity for both gaits and horse breeds except for the crossings per second at the baseline and 75 percentile at walk in the Haflinger horses and 75 percentile at trot in the Noriker horses. The medians of all calculated density values were significantly lower in the GA Haflingers compared to the GG Haflingers (p = 0.012) and also in the AA Norikers compared to the GG Norikers (p = 0.011). Results indicate that the GYS1 mutation reduces the number of functional muscle fibres detected by sEMG measurements even in the absence of overt clinical signs.     

Cricothyroid muscle function and vocal fold stability in exercising horses

OBJECTIVES: To determine (1) if the cricothyroid muscle had respiratory-related electromyographic (EMG) activity that increased with respiratory effort and (2) if bilateral cricothyroid myotomy resulted in vocal fold instability and collapse in exercising horses. STUDY DESIGN: Experimental. ANIMALS: Seven (3 EMG; 4 cricothyroid myotomy) Standardbred horses. METHODS: Three horses exercised on a treadmill at speeds corresponding to the speed that produced maximum heart rate (HR(max)), 75% of maximum heart rate (HR(75%max)), and 50% of maximum heart rate (HR(50%max)) for 60 seconds at each speed while EMG activity of the cricothyroid muscle and nasopharyngeal pressures were measured. Another 4 normal horses were exercised on the treadmill at HR(max) and HR(75%max) for 60 seconds at each speed before and after bilateral cricothyroid myotomy. Upper airway pressures were measured and videoendoscopic examinations were performed and videotaped at each speed. RESULTS: Peak phasic EMBG activity of the cricothyroid muscle was coincident with inspiration and increased with treadmill speed. Bilateral cricothyroid myotomy resulted in vocal fold collapse in all horses. Mean peak inspiratory pressures were significantly more negative compared with control values at both HR(max) and HR(75%max). CONCLUSIONS: Cricothyroid muscle dysfunction may be implicated in vocal fold collapse and likely causes inspiratory airway obstruction in exercising horses. CLINICAL RELEVANCE: Conditions compromising cricothyroid muscle function or motor innervation could result in vocal fold collapse.     

Correlation Between Serum Activity of Muscle Enzymes and Stage of the Estrous Cycle in Italian Standardbred Horses Susceptible to Exertional Rhabdomyolysis

Equine exertional rhabdomyolysis (ER) is a well-recognized clinical syndrome affecting racehorses. Prevalence analysis of ER showed that female sex was a significant risk factor. The aim of this research was to evaluate the differences and correlations in the serum activity of muscle enzymes and the stage of the estrous cycle in ER-susceptible and control (C) mares. Serum muscle enzyme activity before and after exercise and sex hormones were analyzed in the two groups of mares. Ten cyclic ER and 10 cyclic C mares were examined weekly for 4 weeks. During diestrus, ER horses had significantly higher resting and postexercise aspartate aminotransferase (AST) activity, but not creatine kinase (CK) activity, compared with controls; only postexercise AST activity was significantly higher during estrus compared with activity levels in controls. During estrus, 17β-estradiol and AST activity were significantly negatively correlated in the control but not ER mares. Based on our results, further studies should be performed to characterize the presumptive different roles played by sexual hormones in horses susceptible to ER compared with healthy mares.     

Glycogen synthase 1 (GYS1) mutation in diverse breeds with polysaccharide storage myopathy

Background: A missense mutation in the GYS1 gene was recently described in horses with polysaccharide storage myopathy (PSSM).
Objectives: The first objective was to determine the prevalence of the GYS1 mutation in horses with PSSM from diverse breeds. The second objective was to determine if the prevalence of the GYS1 mutation differed between horses diagnosed with PSSM based on grade 1 (typically amylase-sensitive) or grade 2 (typically amylase-resistant) polysaccharide.
Animals: Eight hundred and thirty-one PSSM horses from 36 breeds.
Procedures: Horses with PSSM diagnosed by histopathology of skeletal muscle biopsy samples were identified from the Neuromuscular Disease Laboratory database. Eight hundred and thirty-one cases had blood or tissue that was available for DNA isolation; these 831 cases were genotyped for the GYS1 mutation by restriction fragment length polymorphism.
Results: The PSSM mutation was identified in horses from 17 different breeds. The prevalence of the GYS1 mutation in PSSM horses was high in Draft- (87%) and Quarter Horse-related breeds (72%) and lower in Warmbloods (18%) and other light horse breeds (24%), when diagnosis was based on grade 2 diagnostic criteria. Overall, the PSSM mutation was present in 16% of grade 1 and 70% of grade 2 PSSM horses.
Conclusions and Clinical Importance: GYS1 mutation causes PSSM in diverse breeds and is the predominant form of PSSM in Draft- and Quarter Horse-related breeds. False-positive diagnosis, as well as the possibility of a second glycogenosis in horses with neuromuscular disease (type 2 PSSM), might explain the absence of the GYS1 mutation in horses diagnosed with excessive glycogen accumulation in muscle.     

Incidence of polysaccharide storage myopathy in draft horse-related breeds: a necropsy study of 37 horses and a mule.

Skeletal muscle samples from 38 draft horse-related animals 1-23 years of age were evaluated for evidence of aggregates of glycogen and complex polysaccharide characteristic of equine polysaccharide storage myopathy (EPSSM). Cardiac muscle from 12 of these horses was also examined. Antemortem serum levels of creatine kinase (CK) and aspartate aminotransferase (AST) from 9 horses with EPSSM and 5 horses without EPSSM were compared. Skeletal muscle from 17 horses contained inclusions of periodic acid-Schiff (PAS)-positive, amylase-resistant complex polysaccharide. Similar inclusions were also present in the cardiac muscle of 1 horse. A vacuolar myopathy with aggregates of PAS-positive, amylase-sensitive glycogen was seen in 8 other horses, and these findings are also considered diagnostic for EPSSM. Antemortem serum activities of CK and AST were often higher in EPSSM horses than in horses without EPSSM. Using the presence of amylase-resistant complex polysaccharide as the criterion for diagnosis of EPSSM, the incidence in this population was 45%. Inclusion of horses with aggregates of glycogen but no amylase-resistant complex polysaccharide as representative of the range of pathologic findings in horses with EPSSM resulted in a 66% incidence in this population.     

Packed cell volume and heart rate to predict medical and surgical cases and their short-term survival in horses with gastrointestinal-induced colic

The objective of this prospective case-control study of 125 horses with gastrointestinal tract-induced colic was to determine whether heart rate (HR) and packed cell volume (PCV) can predict surgical versus medical treatment and its short-term survival (time of discharge of the patient). Sixty-four horses were treated medically and 61 surgically (29 small intestinal and 32 large intestinal cases). At admission, both PCV and HR were higher in horses treated surgically than in horses treated medically; however, with longer duration of colic before presentation, the PCV was higher in the small intestinal surgical group only. In addition, both PCV and HR were higher, and the duration of colic was longer in non-survivors compared to survivors. Binary logistic regression demonstrated a significant association between HR and type of treatment, and both HR and PCV were predictive of survival. Simple parameters such as HR and PCV provide useful information for management of colic cases.     

Electromyographic activity of the stylopharyngeus muscle in exercising horses

REASONS FOR PERFORMING STUDY: There is a need to understand the process which leads to failure of recruitment of the stylopharyngeus muscle in clinical cases of nasopharygeal collapse. We therefore studied the timing and intensity of stylopharyngeus muscle activity during exercise in horses. OBJECTIVE: To measure the electromyographic (EMG) activity of the stylopharyngeus muscle in exercising horses and correlate it with the breathing pattern. METHODS: Five horses were equipped with a bipolar fine wire electrode placed on the stylopharyngeus muscle and a pharyngeal catheter. The horses exercised on a treadmill at speeds corresponding to 50 (HRmax50), 75 and 100% of maximum heart rate, and EMG activity of the stylopharyngeus muscle and upper airway pressures were recorded. The EMG activity of the stylopharyngeus muscle was then correlated to the breathing pattern and the activity quantified and reported as a percentage of the baseline activity measured at HRmax50. RESULTS: There was ongoing activity of the stylopharyngeus muscle throughout the breathing cycle; however, activity increased towards the end of expiration and peaked early during inspiration. Tonic activity was present during expiration. Peak, mean electrical and tonic EMG activity increased significantly (P<0.05) with exercise intensity. CONCLUSION: The stylopharyngeus muscle has inspiratory-related activity and tonic activity that increases with speed. POTENTIAL RELEVANCE: The stylopharyngeus muscle is one of a group of upper airway muscles that function to support and maintain the patency of the nasopharynx during inspiration. Failure of recruitment of the stylopharyngeus muscle during exercise is a potential explanation for clinical cases of dorsal pharyngeal collapse, but further work investigating the activity of the stylopharyngeus muscle in horses affected by this disease is needed.     

The apo-enzyme content of aminotransferases in healthy and diseased domestic animals

We investigated the apo-enzyme content of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in clinically normal and ill canine, feline, equine and bovine patients. Aminotransferase activity was measured with and without the addition of exogenous pyridoxal-5'-phosphate (P5P). The amount of apo-enzyme was expressed as the percentage change in aminotransferase activity with the inclusion of P5P. The results of aminotransferase assays without P5P (holo-enzyme activity) were highly correlated to the results obtained with P5P (total enzyme activity) in all four species (Spearman rank correlations > 0.980). The median apo-aminotransferase percentage in clinically normal patients was: 11% ALT and 0% AST in 115 dogs, 7% ALT and -5% AST in 50 cats, 6% AST in 46 horses and 9% AST in 50 cattle. The amount of apo-enzyme did not increase as holo-enzyme activity increased in any of the species. The apo-aminotransferase content was not significantly different in canine, feline and equine patients with diseases affecting the liver, kidney or musculature compared to clinically normal animals. The apo-enzyme content did not differ significantly between specific liver diseases in canine and feline patients.     

Aquapuncture Improves Metabolic Capacity in Thoroughbred Horses

The aim of this study was to evaluate the effect of aquapuncture on muscle enzymes release and cardiovascular and metabolic variables of Thoroughbred horses during and after exercise. Eighteen healthy Thoroughbred horses, submitted to training for competition, were used. The animals were randomly allotted into three groups: ConG (negative control group), ShG (sham aquapuncture or positive control group), and AcuG (treatment aquapuncture group). The treatments were done twice per week, during a 3-week interval. Physical performance was evaluated under field conditions, with two submaximal incremental velocity tests, separated by a 3-week period. The tests included evaluation of cardiovascular (V₂₀₀) and metabolic (V_La4) capacities and measurement of creatine kinase (CK) and aspartate aminotransferase (AST) before and after exercise. V₂₀₀ values were not different between times or groups. V_La4 values were statistically greater after treatment for the AcuG group compared with pretreatment. CK concentrations were statistically greater after exercise for all groups, and AST concentrations were not different between times or groups. Aquapuncture influenced neither cardiovascular capacity nor the release of muscle enzymes; however, it did improve metabolic capacity of Thoroughbred horses.     

A glycogen synthase 1 mutation associated with equine polysaccharide storage myopathy and exertional rhabdomyolysis occurs in a variety of UK breeds

Reasons for performing study: A glycogen synthase (GYS1) mutation has been described in horses with histopathological evidence of polysaccharide storage myopathy (PSSM) in the USA. It is unknown whether the same mutation is present in horses from the UK. Objectives: To determine whether the GYS1 mutation occurs in UK horses with histopathological evidence of PSSM and exertional rhabdomyolysis. Hypothesis: The R309H GYS1 mutation is present in a variety of UK horse breeds and that the mutation is commonly associated with exertional rhabdomyolysis. Methods: DNA was extracted from 47 muscle or blood samples from UK horses with histories of exertional rhabdomyolysis in which muscle biopsy diagnosis had been pursued. The proportions of GYS1 mutation positive cases were compared among histopathologically defined groups. In addition, breeds that carried the GYS1 mutation were identified from a total of 37 grade 2 (amylase‐resistant) PSSM cases. Results: Of 47 horses with exertional rhabdomyolysis in which a muscle biopsy diagnosis was pursued, 10 (21%) carried the GYS1 mutation. The mutation was only found in horses with grade 2 PSSM (i.e. not in horses with normal, idiopathic myopathy or grade 1 PSSM biopsy samples). In total, the GYS1 mutation was found in 24/37 (65%) of grade 2 PSSM cases. A variety of breeds, including Quarter Horse, Appaloosa, Warmblood, Connemara‐cross, Cob, Polo Pony and Thoroughbred cross carried the mutation. Conclusions: The GYS1 mutation is an important cause of exertional rhabdomyolysis of UK horse breeds but does not account for all forms of PSSM. Potential relevance: Genotyping is recommended in cases of exertional rhabdomyolysis, prior to or in combination with, muscle biopsy. However a significant proportion of horses with histopathological evidence of PSSM and/or exertional rhabdomyolysis have different diseases.     

Sarcoplasmic Reticulum Ca2+-ATPase Activity and Glycogen Content in Various Fiber Types after Intensive Exercise in Thoroughbred Horses

To find a new parameter indicating muscle fitness in Thoroughbred horses, we examined time-dependent recovery of glycogen content and sarcoplasmic reticulum (SR) Ca²⁺-ATPase activity of skeletal muscle after intensive treadmill running. Two repeated 50-sec running sessions (13 m/sec) were performed on a flat treadmill (approximately 90%VO₂max). Muscle samples of the middle gluteal muscle were taken before exercise (pre) and 1 min, 20 min, 60 min, and 24 hr after exercise. Muscle fiber type composition was determined in the pre muscle samples by immunohistochemical staining with monoclonal antibody to myosin heavy chain. SR Ca²⁺-ATPase activity of the muscle and glycogen content of each muscle fiber type were determined with biochemical analysis and quantitative histochemical staining, respectively. As compared to the pre value, the glycogen content of each muscle fiber type was reduced by 15–27% at 1 min, 20 min, and 60 min after the exercise and recovered to the pre value at 24 hr after exercise test. These results indicate that 24 hr is enough time to recover glycogen content after short-term intensive exercise. The mean value of the SR Ca²⁺-ATPase activity showed a slight decrease (not significant) immediately after exercise, and complete recovery at 60 min after exercise. There were no significant relationship between the changes in glycogen content of each muscle fiber type and SR Ca²⁺-ATPase. Although further studies are needed, SR Ca²⁺-ATPase is not a useful parameter to detect muscle fitness, at least in Thoroughbred horses.     

The evaluation of selected parameters of cellular nonspecific immunity in normal and allergic horses

The main aim of this study was to compare selected nonspecific immunity parameters in 14 allergic and 12 healthy horses. Each animal was assessed according to the following parameters: in vitro functional capacity of phagocytic cells using the nitro blue tetrazolium chloride reduction test, both spontaneous (NBT) and zymozan stimulated (NBTs), and ingestion capacity of phagocytic cells using a phagocytic index test (IF) and percentage of phagocytosing neutrophils activity (%KF). Differences were demonstrated between the group of allergic horses, especially with severe allergy symptoms, and healthy horses in NBTs values, with higher values in healthy horses. The values of the phagocytic index were significantly higher in horses with allergy.     

Neuronal nitric oxide synthase is heterogeneously distributed in equine myofibers and highly expressed in endurance trained horses

Mammalian skeletal muscle expresses splice variants of neuronal nitric oxide synthase (nNOS). Skeletal muscles have a metabolically heterogeneous population of myofibers, and fiber composition in equine skeletal muscle is correlated with athletic ability in endurance events. In this study, we investigated whether nNOS expression in equine skeletal muscle is related to fiber type and endurance training. Biopsy samples obtained from the gluteus medius of sedentary- (SH) and endurance-trained (TH) horses were examined for the electrophoretic mobility of myosin heavy chain (MHC) and NOS activity. Serial tissue cross-sections were stained for myosin ATPase and nicotinamide adenine dinucleotide (NADH) reductase, and also immunostained for nNOS. The gluteus medius of TH had higher levels of nNOS expression and activity when compared to muscle from SH. In SH, nNOS was restricted to the subsarcolemmal area while in TH nNOS was also present at cytoplasmic sites. A splice variant of nNOS was heterogeneously distributed among the different myofibers, its expression being higher in fast-oxidative-glycolytic type IIA fibers than in fast-glycolytic type IIX fibers and absent in slow-twitch type I fibers. Trained horses had a significantly higher relative content of type IIA fibers, a greater oxidative capacity, and a lower percentage of type IIX fibers when compared with SH. The differences in muscle fiber typing between the 2 groups of horses reflected alterations that probably resulted from the endurance-training program. Overall, these results show that nNOS is differentially expressed and localized in the gluteus medius according to the fiber type and the athletic conditioning of the horses.     

The Effect of Training on Yili Horse 1 600 m Test Results and Heart Rate Variability

【Objective】 The objective of this study was to analyze the effect of different training stages on the performance and heart rate variability(HRV) of Yili horses and to provide data for reference in the conditioning training of 1 600 m distance Yili horses.【Method】 Eight 3-year-old Yili horses (stallions) were selected as test subjects.A 3-month speed specific performance training program was conducted, and a speed test race was organized in the last week of each month of training, and HRV was collected before, immediately after, 0.5 h after, and 1 h after 1 600 m test race.Among them, the time-domain indexes included the mean of all R-R intervals (Mean RR), the standard deviation of all R-R intervals (SDNN), the mean heart rate (Mean HR), the root mean square of the difference between adjacent R-R intervals (RMSSD), the number of adjacent R-R intervals with a difference greater than 50 ms (NN50), and the percentage of adjacent R-R intervals with a difference greater than 50 ms to the total number of heartbeats (pNN50).Frequency domain metrics included very low frequency (VLF), low frequency power (LF), high frequency power (HF) and nonlinear metrics:standard deviation (Y) of all R-R-spacing (SD1), standard deviation (X) of all R-R-spacing (SD2).Consequently, the variability of HRV indexes in horses at different training stages was analyzed.【Result】 The race time in the post training period in 1 600 m speed test race of Yili horses was significantly lower than that in the early training period (P<0.05), and Mean RR, NN50 and pNN50 in the middle and post training periods were significantly lower than that in the early training period (P<0.05).Mean HR in the beginning of training was significantly lower than that in post training (P<0.05).VLF and LF in the end of training were significantly lower than that in the beginning of training (P<0.05).【Conclusion】 Under the conditions of this test, the HRV indexes of 1 600 m test race of Yili horses in different conditioning training stages were analyzed.The outcomes demonstrated that the types of neural activity in Yili horses at the beginning, middle and end of training presented some differences, showing changes of increased sympathetic excitability and decreased parasympathetic excitability, and the horses' athletic performance improved.Therefore, HRV could be considered as an effective tool to evaluate the training load and intensity of Yili horses during training.     

Muscle characteristics in young Norwegian-Swedish Coldblooded Trotters and associations with breeding index, body size and early training

Reasons for performing study: The trotting speed of Norwegian‐Swedish Coldblooded Trotters (NSCTs) is, although inferior to Standardbred speeds, gradually increasing. This development is mainly attributed to breeding progresses. Characteristics of the middle gluteal muscle have recently been described in a small number of these horses but the actual spread in muscle parameters within the population is unknown. Objectives: To describe the characteristics of the middle gluteal muscle and their naturally occurring variability in young NSCT horses and investigate possible association with body size, individual breeding index and early training. Methods: Biopsies of the middle gluteal muscle of 144 NSCT horses born in 2005 were collected at age 17–24 months. The horses were randomly selected by stratified sampling with gender and paternal progeny as stratification factors. The muscle samples were analysed for fibre type composition, fibre type areas, capillary supply and the activity of the key metabolic enzymes citrate synthase, 3‐OH‐acyl‐CoA‐dehydrogenase and lactate dehydrogenase. Correlation analysis including muscle parameters and body size, individual breeding index and early training was performed. Results: Large interindividual differences were identified in muscle characteristics. Male horses had higher percentages of type IIA fibres and lower percentages of type IIX fibres than age‐matched females. Fibre type areas followed the order type IIX>IIAX>IIA>I, while the capillary supply per fibre type area increased in the opposite order. No gender‐related differences were found in fibre type areas or capillary supply. Height was correlated to the relative area of type I fibres. Early training was associated with increased oxidative capacity and increased percentage of type IIA fibres. The lowest breeding index was found in untrained horses. Conclusions: Muscle characteristics differ among young NSCT horses, but the innate oxidative capacity seems to be lower than in Standardbreds of comparable age. Early training may influence on these parameters.     

Emotional Response to Novelty and to Expectation of Novelty in Young Race Horses

The aim of this study was to estimate the emotional response to novelty and to expectation of novelty in young race horses. The novelty in this study was the first training on an automated horse walker at a new training center. To estimate the level of emotionality in horses, the telemetric measurement of heart rate (HR) was used. A hypothesis was developed that expectation of novelty can be as exciting for horses as a novelty test. In this study, 40 horses were studied just before and then during their first walk on an automated horse walker. They were divided into four groups, with 10 horses in each group. These groups were as follows: (1) 1.5-year-old Thoroughbred colts, (2) 1.5-year-old Thoroughbred fillies, (3) 2.5-year-old Purebred Arabian colts, and (4) 2.5-year-old Purebred Arabian fillies. HR was measured at rest before exercise, during handling and moving the horse from the stable, while walking on the automated horse walker for about 20 minutes, while moving the horse from the walker to the stable, and at rest after exercise. HR response to the anticipation of novelty was higher in colts than in fillies, particularly in the group of Thoroughbreds.     

Comparison of plasma biochemical parameters in Thoroughbred and Purebred Arabian horses during the same-intensity exercise

The objective of the present study was to compare changes of blood parameters induced by the same work and performed in the same conditions in two racehorse breeds, Thoroughbred and Purebred Arabian. The effect of moderate-intensity exercise was studied in 20 stallions--ten Thoroughbreds, aged 2-3 years and the same number of Purebred Arabians, 3-4 years old. All the horses were administrated the same effort test consisting in 1200 m gallop at a speed typical of the daily training sessions. Three jugular venous blood samples were collected for each horse: at rest, just after the end of the gallop and after 30-minute rest. In the gathered blood, a hemoglobin (Hb) concentration was determined as well as plasma level of glucose (Glc), triacylglycerols (TG), glycerol, free fatty acids (FFA), total plasma proteins (TP) and the activity of creatine kinase (CK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST). In the Arabian horses, an increase in levels of TP, glycerol, FFA and CK activity measured just after exercise was higher than that in Thoroughbreds. Similarly, after a 30-minute rest, a post-exercise rise of TP, AST, glycerol and FFA proved to be higher in the Arabian horses compared to that in the Thoroughbreds. Only TG plasma concentration measured 30 minutes following the effort was significantly lower in the Arabian horses than in Thoroughbreds. It can be concluded that the Thoroughbred horses adapted better to the effort test applied in this study as compared to the Purebred Arabian horses. The parameters related to lipid metabolism proved to be the most sensitive indicators of breed differences in relation to moderate-intensity exercise.     

Preliminary Study on Age- and Sex-Dependent Alterations in the Composition of Skeletal Muscle Fibers of Brasileiro de Hipismo Horses

The aim of this work was to characterize the distribution of myofibers in the gluteus medius muscle of inactive horses of the Brasileiro de Hipismo (BH) breed at different ages by means of histochemical analyses, according to sex and depth of the biopsy. A total of 78 inactive horses (9 castrated males, 35 stallions, and 34 females) of the BH breed, aged 1 to 4 years, were used. A percutaneous muscle biopsy was obtained with a 6.0-mm Bergström-type needle, which allowed the removal of muscle fragments at depths of 20 and 60 mm. Myofiber types were determined based on myofibrillar adenosine triphosphatase (mATPase) and nicotinamide dinucleotide tetrazolium reductase (NADH-TR) techniques. Morphometry of the fibers was determined based on cross-sectional area (CSA), mean frequency (F), and relative cross-sectional area (RCSA). The current study demonstrated that BH horses 3 and 4 years of age show a greater percentage of, and area occupied by, type IIA fibers and lower percentage of type IIX fibers in the gluteus medius muscle compared with horses 1 and 2 years of age. No difference was found between sexes in the frequency of and area occupied by the different fiber types at any of the depths and ages studied. In this study, females showed a greater CSA for all fibers in comparison with males, at 1 year of age. The results of the current study indicate that the gluteus medius muscle of inactive BH horses shows modifications in its structural and biochemical composition during the growth of the animals, leading to a better oxidative capacity.     

Epidemiological and genetic study of exertional rhabdomyolysis in a Warmblood horse family in Switzerland

Reasons for performing study: Exertional rhabdomyolysis (ER) and its familial basis in Warmblood horses is incompletely understood. Objectives: To describe the case details, clinical signs and management of ER‐affected Warmblood horses from a family with a high prevalence of ER, to determine if histopathological signs of polysaccharide storage myopathy (PSSM) and the glycogen synthase (GYS1) mutation are associated with ER in this family, and to investigate potential risk factors for development of ER. Methods: A family consisting of a sire with ER and 71 of his descendants was investigated. History of episodes of ER, husbandry, feeding and use was assessed by interviewing horse owners using a standardised questionnaire. All horses were genotyped for GYS1. In 10 ER‐affected horses, muscle histopathology was evaluated. Results: Signs of ER were reported in 39% of horses and 51% of the entire family possessed the GYS1 mutation. Horses possessing the GYS1 mutation had a 7.1‐times increased risk for developing ER compared to those with the normal genotype (95% confidence interval [CI] 2.37–21.23, P = 0.0005). All muscle samples from horses in the family with ER showed polysaccharide accumulation typical for PSSM, amylase‐resistant in 9/10 cases. There was evidence (odds ratio 5.6, CI 1.00–31.32, P = 0.05) that fat or oil feeding improved clinical signs of ER. No other effects of environmental factors associated with clinical signs of ER were identified. Conclusion and potential relevance: PSSM associated with the GYS1 mutation is one identifiable cause of ER in Warmblood horses. Signs of ER are not always manifest in GYS1 positive horses and there are also other causes for ER in Warmblood horses. Breeding animals with the GYS1 mutation results in a high prevalence of ER due to its dominant mode of inheritance.     

The Interplay of Genetics,Exercise, and Nutrition in Polysaccharide Storage Myopathy

Polysaccharide storage myopathy (PSSM), identified in 1992 in a subset of horses with exertional rhabdomyolysis, is a glycogenosis characterized by amylase-resistant polysaccharide in a small number of skeletal muscle fibers along with 1.5 to 4 times normal muscle glycogen. Extensive biochemical and physiological analyses failed to identify defects in glycogenolysis and glycolysis. In 2008, a genome-wide association analysis detected a locus on equine chromosome 10 that was strongly associated with the PSSM in Quarter Horses. Glycogen synthase 1 (GYS1), which encodes the skeletal muscle isoform of glycogen synthase (GS), was a strong candidate gene for PSSM based on its location on equine chromosome 10. Sequencing of the GYS1 gene in PSSM and control Quarter Horses identified only one single base-pair change that resulted in an amino acid substitution in the GS enzyme. Mean GS activity was higher in PSSM than control muscle homogenates in both the presence and absence of the allosteric activator glucose 6-phosphate, suggesting that the GS enzyme in horses with PSSM is constitutively active. High-grain diets increase serum insulin concentrations which further act to stimulate GS activity. An restriction fragment length polymorphism assay for the GYS1 mutation showed that 10% of the Quarter Horse breed and a minimum of 20 other breeds have the GYS1 mutation. Muscle biopsies obtained after 20 minutes of aerobic exercise revealed much higher inosine monophosphate concentrations and lower adenosine monophosphate in whole muscle and single fibers from PSSM as compared with control horse muscle. Thus, the GYS1 mutation responsible for PSSM seems to cause an energy imbalance exacerbated by high-grain diets, which results in adenine nucleotide degradation in individual muscle fibers of horses with PSSM during submaximal exercise.