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Gross morphology of healthy and degenerated intervertebral discs (IVDs) is largely similar in horses as in dogs and humans. For further comparison, the biochemical composition and the histological and biochemical changes with age and degeneration were analyzed in 41 warmblood horses. From 33 horses, 139 discs and 2 fetal vertebral columns were evaluated and scored histologically. From 13 horses, 73 IVDs were assessed for hydration, DNA, glycosaminoglycans, total collagen, hydroxyl-lysyl-pyridinoline, hydroxylysine, and advanced glycation end-product (AGE) content. From 7 horses, 20 discs were assessed for aggrecan, fibronectin, and collagen type 1 and 2 content. Histologically, tearing of the nucleus pulposus (NP) and cervical annulus fibrosus (AF), and total histological score (tearing and vascular proliferation of the AF, and chondroid metaplasia, chondrocyte-like cell proliferation, presence of notochordal cells, matrix staining, and tearing of the NP) correlated with gross degeneration. Notochordal cells were not seen in IVDs of horses. Age and gross degeneration were positively correlated with AGEs and a fibrotic phenotype, explaining gross degenerative changes. In contrast to dogs and humans, there was no consistent difference in glycosaminoglycan content and hydration between AF and NP, nor decrease of these variables with age or degeneration. Hydroxylysine decrease and collagen 1 and AGEs increase were most prominent in the NP, suggesting degeneration started in the AP. In caudal cervical NPs, AGE deposition was significantly increased in grossly normal IVDs and total collagen significantly increased with age, suggesting increased biomechanical stress and likelihood for spinal disease in this part of the vertebral column.

Background: Changes in cardiovascular parameters, including blood pressure (BP) and cardiac anatomical dimensions, are an inconsistent feature of the equine metabolic syndrome. The order in which these changes arise is unknown., Objectives: Determine the order in which EMS-associated changes in cardiovascular parameters arise., Animals: Twenty Shetland pony mares., Methods: High-energy (HE) diet mares were fed 200% of net energy requirements for 1 (n = 3) or 2 (n = 7) consecutive diet-years, with 17 weeks of hay-only between years. Noninvasive BP measurements and echocardiograms were performed during both years. Resting 24-hour ECGs and measurements of autonomic tone (splenic volume and packed cell volume [PCV]) were performed at the end of diet-year 1. Results were compared to control mares receiving a maintenance diet for 1 (n = 7) or 2 (n = 3) consecutive years., Results: In year 1, HE mares had significantly higher values than control mares for mean relative left ventricular wall thickness (P = .001). After 2 diet-years, mean systolic (P = .003), diastolic (P < .001) and mean arterial BP (P = .001), heart rate (HR; P < .001), and mean left ventricular wall thickness (P = .001) also were significantly increased in HE compared to control mares. No pathological arrhythmias or differences in splenic volume or PCV were detected., Conclusions and Clinical Importance: Ingesting a HE diet first induced minor changes in BP, and progressed to left-sided cardiac hypertrophy in Shetland pony mares. These findings are of interest given the increasing incidence of obesity in horses., (© 2021 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.)

Saliva is a matrix which may act as a vector for pathogen transmission and may serve as a possible proxy for SARS-CoV-2 contagiousness. Therefore, the possibility of detection of intracellular SARS-CoV-2 in saliva by means of fluorescence in situ hybridization is tested, utilizing probes targeting the antisense or sense genomic RNA of SARS-CoV-2. This method was applied in a pilot study with saliva samples collected from healthy persons and those presenting with mild or moderate COVID-19 symptoms. In all participants, saliva appeared a suitable matrix for the detection of SARS-CoV-2. Among the healthy, mild COVID-19-symptomatic and moderate COVID-19-symptomatic persons, 0%, 90% and 100% tested positive for SARS-CoV-2, respectively. Moreover, the procedure allows for simultaneous measurement of viral load ('presence', sense genomic SARS-CoV-2 RNA) and viral replication ('activity', antisense genomic SARS-CoV-2 RNA) and may yield qualitative results. In addition, the visualization of DNA in the cells in saliva provides an additional cytological context to the validity and interpretability of the test results. The method described in this pilot study may be a valuable diagnostic tool for detection of SARS-CoV-2, distinguishing between 'presence' (viral load) and 'activity' (viral replication) of the virus. Moreover, the method potentially gives more information about possible contagiousness., Competing Interests: WJBvW and IDW have declared that no competing interests exist. GJJ and MW have read the journal’s policy and these authors of this manuscript have the following competing interests: employment by the commercial company Biotrack and patents issued (WO 2010/040371 A1, EP 08874964.3). This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Background: In Warmblood horses, degenerative joint disease is involved in cervical malformation and malarticulation (CVM). The degree of contribution of articular process joint (APJ) osteochondrosis (OC) is not clear., Objectives: (a) To explore the presence of predilection sites for APJ OC in cervical and cranial thoracic vertebral columns of Warmblood foals and (b) to examine the correlation of such a site with the predilection site of CVM., Study Design: Case series., Methods: Seven hundred APJ facets of C2 to T2 of 29 foals (11 months gestation to 12 months [median age 7 days; range 365 days; 95% confidence interval [95% CI] 2-47 days]) were examined for OC and prevalence between joints, and the predilection site for CVM and the cranial cervical vertebral column were evaluated., Results: About 20.6% of facets revealed OC. There was no predilection site. Prevalence decreased with age up to 1 year (odds ratio [OR] 0.997; (95% CI 0.975-0.998)) but not up to 5 months. Severity increased with age in all age ranges (up to 1 year OR 1.023; 95% CI 1.005-1.049; >1-5 months, OR 1.203; 95% CI 1.014e+00-1.921; up to 1 month, OR 1.114; 95% CI 1.041-1.228). Highest prevalence was in cranial facets of the cervical and cervical-thoracic joints and in caudal facets of the thoracic joint up to 1 year and up to 1 month (OR 0.364; 95% CI 0.170-0.745, OR 0.434; 95% CI: 0.235-0.782, OR 7.665; 95% CI: 1.615-66.553 and OR 0.400; 95% CI 0.170-0.880, OR 0.351; 95% CI 0.172-0.700, OR 5.317; 95% CI 1.098-44.344 respectively)., Main Limitations: Two-thirds of the foals were less than 1 month of age., Conclusions: Articular process joint OC in Warmblood foals is common and is not more prevalent at CVM predilection sites, suggesting that abnormalities of enchondral ossification may not be major contributors to CVM., (© 2020 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.)

Background: Variation in equine caudal cervical spine morphology at C6 and C7 has high prevalence in Warmblood horses and is suspected to be associated with pain in a large mixed-breed group of horses. At present no data exist on the relationship between radiographic phenotype and clinical presentation in Warmblood horses in a case-control study., Objectives: To establish the frequency of radiographically visible morphologic variation in a large group of Warmblood horses with clinical signs and compare this with a group without clinical signs. We hypothesised that occurrence of morphologic variation in the case group would not differ from the control group, indicating there is no association between clinical signs and morphologic variation., Study Design: Retrospective case-control., Methods: Radiographic presence or absence of morphologic variation of cervical vertebrae C6 and C7 was recorded in case (n = 245) and control horses (n = 132). Case and control groups were compared by univariable Pearson's Chi-square and multivariable logistic regression for measurement variables age, sex, breed, degenerative joint disease and morphologic variation at C6 and C7. Odds ratio and confidence intervals were obtained. A P≤0.05 was considered statistically significant., Results: Morphologic variation at C6 and C7 (n = 108/377 = 28.6%; Cases 58/245 = 23.7%; Control 50/132 = 38%) was less frequent in horses with clinical signs in univariable testing (OR 0.48, 95% CI 0.3-0.8, P = 0.001). Age, sex, breed and degenerative joint disease were not retained in the final multivariable logistic regression step whereas morphologic variation remained significantly less present in horses with clinical signs., Main Limitations: Possible demographic differences between equine clinics., Conclusions: Morphologic variation in the caudal cervical spine was detected more frequently in horses without clinical signs. Therefore, radiographic presence of such variation does not necessarily implicate the presence of clinical signs., (© 2019 The Authors Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.)

Morphology of the equine cervical intervertebral disc is different from that in humans and small companion animals and published imaging data are scarcely available. The objectives of this exploratory, methods comparison study were (a) to describe MRI features of macroscopically nondegenerated and degenerated intervertebral discs (b) to test associations between spinal location and macroscopic degeneration or MRI-detected annular protrusion and between MRI-detected annular protrusion and macroscopic degeneration, and (c) to define MRI sequences for characterizing equine cervical intervertebral disc degeneration. Ex vivo MRI of intervertebral discs was performed in 11 horses with clinical signs related to the cervical region prior to macroscopic assessment. Mixed-effect logistic regression modeling included spinal location, MRI-detected annular protrusion, and presence of macroscopic degeneration with "horse" as random effect. Odds ratio and 95% confidence interval were determined. Reduced signal intensity in proton density turbo SE represented intervertebral disc degeneration. Signal voids due to presence of gas and/or hemorrhage were seen in gradient echo sequences. Presence of macroscopic intervertebral disc degeneration was significantly associated with spinal location with odds being higher in the caudal (C5 to T1) versus cranial (C2 to C5) part of the cervical vertebral column. Intervertebral discs with MRI-detected annular protrusion grades 2-4 did have higher odds than with grade 1 to have macroscopic degeneration. It was concluded that MRI findings corresponded well with gross macroscopic data. Magnetic resonance imaging of the equine cervical intervertebral disc seems to be a promising technique, but its potential clinical value for live horses needs to be explored further in a larger and more diverse population of horses., (© 2019 The Authors. Veterinary Radiology & Ultrasound published by Wiley Periodicals, Inc. on behalf of American College of Veterinary Radiology.)

Congenital portosystemic shunts in foals are rare and only a small number of cases have been described. Detailed description of the course of the shunt is lacking in earlier reports. This is the first detailed description of a computed tomography angiography (CTA) displaying an extra-hepatic splenocaval shunt. A 1-month old colt showing increasing signs of dullness, ataxia, circling, lip-smacking and coordination problems was presented. Hyperammonemia was detected and abdominal CTA revealed an extra-hepatic portocaval shunt. During surgery, ligation of the abnormal vessel could not be achieved, and the foal was euthanized because of complications during surgery. CTA provided a detailed overview of portal vasculature. If a portosystemic shunt is suspected in a foal, CTA can be used to confirm the diagnosis and for surgical planning.

Phosphorus (P) deficiency and hypophosphatemia are believed to be associated with muscle function disturbances in dairy cows, particularly around parturition. The objective of this study was to determine the effect of dietary P deprivation during late gestation and early lactation on muscle P homeostasis and muscle function in periparturient dairy cows. Thirty-six multiparous dairy cows in late gestation were randomly assigned either to undergo dietary P depletion or to be offered a diet with adequate P content from 4 wk before to 4 wk after parturition. Phosphorus-deficient rations for dry and lactating cows contained 0.15 and 0.20% P on a dry matter basis, respectively. Blood and muscle tissue for biopsy were obtained and electromyographic examinations were conducted on biceps femoris and intercostal muscles in regular intervals throughout the study. Muscle tissue was analyzed for the total P, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, creatine phosphate, and tissue water content. Dietary P deprivation resulted in a pronounced and sustained decline of the plasma phosphate concentration, reaching a nadir at calving with mean values below 1.5 mg/dL and remaining below 2.0 mg/dL during the first 4 wk of lactation. Hypophosphatemia was not associated with signs of clinically apparent muscle weakness or disturbed muscle function and was not associated with a decline in the content of any of the studied P-containing compounds in muscle tissue. Accordingly, no association between plasma phosphate concentration and muscle tissue P content was found. Electromyographic examination identified subclinical effects on motor unit action potentials that are indicative of disturbed neuromuscular functionality. Increasing occurrence of pathologic spontaneous activity possibly resulting from membrane instability of nerve or muscle cells and suggestive of myopathy was also recorded as P deprivation progressed. These effects were predominantly observed in intercostal and to a lesser degree biceps femoris muscles. Electromyographic parameters affected by P deprivation were found to be associated primarily with the plasma phosphate and to a lesser extent with the amounts of energy storing P-containing compounds contained in muscle tissue. These results indicate that prolonged and pronounced dietary P deprivation in transition dairy cows leads to marked sustained hypophosphatemia without altering the muscle tissue P homeostasis or causing clinically apparent muscle function disturbances., (The Authors. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

Equine intervertebral disc degeneration is thought to be rare and of limited clinical relevance, although research is lacking. To objectively assess pathological changes of the equine intervertebral disc and their clinical relevance, description of the normal morphology and a practical, biologically credible grading scheme are needed. The objectives of this study are to describe the gross and histological appearance of the equine intervertebral discs and to propose a grading scheme for macroscopic degeneration. Spinal units from 33 warmblood horses were grossly analyzed and scored. Of the 286 intervertebral discs analyzed, 107 (37%) were assigned grade 1 and grade 2 (considered normal) and were analyzed histologically. A nucleus pulposus and an annulus fibrosus could be identified macroscopically and histologically. Histologically, the nucleus pulposus was composed of a cartilaginous matrix and the annulus fibrosus of parallel collagenous bands. A transition zone was also histologically visible. Intra- and inter-observer reliability scores were high for all observers. Higher grades were associated with greater age. Gross changes associated with equine intervertebral disc degeneration (grades 3-5)-that is, yellow discoloration, cleft formation (tearing), and changes in consistency of the nucleus pulposus-were largely similar to those in humans and dogs and were most prevalent in the caudal cervical spine. Equine intervertebral disc degeneration was not associated with osteophyte formation. Changes of the vertebral bone were most common in the thoracolumbar spine but were not correlated with higher grades of intervertebral disc degeneration. Thus, changes of the vertebral bone should be excluded from grading for equine intervertebral disc degeneration.

Reasons for Performing Study: Reference values for quantitative electromyography (QEMG) in shoulder and hindlimb muscles of horses are limited., Objectives: To determine normative data on QEMG analysis of supraspinatus (SS), infraspinatus (IS), deltoideus (DT) and biceps femoris (BF) muscles., Study Design: Experimental observational study and retrospective case series., Methods: Seven adult healthy Royal Dutch sport horses underwent quantitative motor unit action potential analysis of each muscle using commercial electromyography equipment. Measurements were made according to published methods. One-way ANOVA was used to compare quantitative motor unit action potential variables between muscles, with post hoc testing according to Bonferroni, with significance set at P<0.05. The QEMG and clinical information from horses with lower motor neuron disorders (n = 7) or myopathy (n = 4) were summarised retrospectively., Results: The 95% confidence intervals of duration, amplitude, phases, turns, area and size index of quantitative motor unit action potential were 8.7-10.4 ms, 651-867 μV, 3.2-3.7, 3.7-4.7, 1054-1457 μV·ms and 1.1-1.5 for SS, 9.6-11.0 ms, 779-1082 μV, 3.3-3.7, 3.8-4.7, 1349-2204 μV·ms and 1.4-1.9 for IS, 6.0-9.1 ms, 370-691 μV, 2.9-3.7, 2.8-4.5, 380-1374 μV·ms and 0.3-1.3 for DT and 5.7-7.8 ms, 265-385 μV, 2.7-3.2, 2.6-3.1, 296-484 μV·ms and 0.2-0.5 for BF, respectively. Mean duration, amplitude, number of phases and turns, area and size index were significantly (P<0.01) higher in SS and IS than in DT and BF muscles. In addition, 4 of 7 normal horses had >15% polyphasic motor unit action potentials in SS and IS muscles., Conclusions: Differences between muscles should be taken into account when performing QEMG in order to be able to distinguish normal horses from horses with suspected neurogenic or myogenic disorders. These normal data provide the basis for objective QEMG assessment of shoulder and hindlimb muscles. Quantitative electromyography appears to be helpful in diagnosing neuropathies and discriminating these from myopathies., (© 2015 EVJ Ltd.)

This review discusses the scope of using (quantitative) electromyography (EMG) in diagnosing myopathies and neuropathies in equine patients. In human medicine, many EMG methods are available for the diagnosis, pathophysiological description and evaluation, monitoring, or rehabilitation of patients, and some of these techniques have also been applied to horses. EMG results are usually combined with other neurophysiological data, ultrasound, histochemistry, biochemistry of muscle biopsies, and clinical signs in order to provide a complete picture of the condition and its clinical course. EMG technology is commonly used in human medicine and has been subject to constant development and refinement since its introduction in 1929, but the usefulness of the technique in equine medicine is not yet widely acknowledged. The possibilities and limitations of some EMG applications for equine use are discussed., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: In equine laminitis, the deep digital flexor muscle (DDFM) appears to have increased muscle force, but evidence-based confirmation is lacking., Objectives: The purpose of this study was to test if the DDFM of laminitic equines has an increased muscle force detectable by needle electromyography interference pattern analysis (IPA)., Animals and Methods: The control group included six Royal Dutch Sport horses, three Shetland ponies and one Welsh pony [10 healthy, sound adults weighing 411 ± 217 kg (mean ± SD) and aged 10 ± 5 years]. The laminitic group included three Royal Dutch Sport horses, one Friesian, one Haflinger, one Icelandic horse, one Welsh pony, one miniature Appaloosa and six Shetland ponies (14 adults, weight 310 ± 178 kg, aged 13 ± 6 years) with acute/chronic laminitis. The electromyography IPA measurements included firing rate, turns/second (T), amplitude/turn (M) and M/T ratio. Statistical analysis used a general linear model with outcomes transformed to geometric means., Results: The firing rate of the total laminitic group was higher than the total control group. This difference was smaller for the ponies compared to the horses; in the horses, the geometric mean difference of the laminitic group was 1.73 [geometric 95% confidence interval (CI) 1.29-2.32], and in the ponies this value was 1.09 (geometric 95% CI 0.82-1.45)., Conclusion and Clinical Relevance: In human medicine, an increased firing rate is characteristic of increased muscle force. Thus, the increased firing rate of the DDFM in the context of laminitis suggests an elevated muscle force. However, this seems to be only a partial effect as in this study, the unchanged turns/second and amplitude/turn failed to prove the recruitment of larger motor units with larger amplitude motor unit potentials in laminitic equids.

Reasons for Performing Study: Atrial fibrillation is a common equine arrhythmia. Quinidine alone, or with digoxin are common treatments. Studies on outcome in Warmblood populations in which duration of the AF is often unknown are limited., Objectives: To identify the factors that are associated with the success of full treatment cardioversion with oral medication, and establish whether there are differences in these factors between institutions., Study Design: Retrospective case series using patient records of Equine University Clinic of Utrecht University and Rossdales Equine Hospital, Newmarket., Methods: Forty-nine horses treated with quinidine were identified (29 Warmbloods, 20 Thorougbreds, 1 Anglo-Arabian). Details of signalment, history, duration physical examination and echocardiography including left atrial size and presence of mitral regurgitation were retrieved. Clinical details including mean weight, age and left atrial size were compared between clinics using independent samples t test. Association between variables and cardioconversion were evaluated in a backwards logistic regression using Akaike's information criterium (AIC) and odds ratios were calculated. Factors were sex, clinic, breed, mitral regurgitation, duration and poor performance. Covariates were age, weight and the size of the left atrium. Significance was set at 0.05., Results: Fifty-one horses (mean age 8.8 s.d. 4.5 years) were treated with quinidine sulfate, 18 also received digoxin. Eighty per cent converted to sinus rhythm. In 8 horses the known duration was less than 3 months. The only factor associated with successful treatment was the use of digoxin in combination with quinidine sulfate (odds ratio 12.4; 95% CI 2.61 and 91.85 according to AIC analysis)., Conclusions: In this retrospective case series, there is much potential for bias in the data; however, the use of digoxin in addition to quinidine was associated with improved conversion rates regardless of breed even though AF duration was unknown in most horses. Ethical animal research: Research ethics committee oversight not currently required by this conference: retrospective study of clinical records. Explicit owner informed consent for inclusion of animals in this study was not stated., Source of Funding: None. Competing interests: None declared., (© 2015 The Author(s). Equine Veterinary Journal © 2015 EVJ Ltd.)

Reasons for Performing Study: In cases of laminitis, an increased muscle force or contracture of the deep digital flexor muscle (DDFM) is suggested, but evidence-based research is lacking., Objectives: To test if the DDFM of laminitic equines shows an increased muscle force detectable by needle-EMG including Interference Pattern Analysis (IPA)., Study Design: Cross-sectional study., Methods: Three groups consisted of Group 0 (control): 6 Royal Dutch Sport horses, 3 Shetland ponies and one Welsh pony (healthy, sound adults, mean ± s.d. weight 411 ± 217 kg). Group 1: 3 Royal Dutch Sport horses, one Friesian, one Haflinger, one Icelandic horse, 2 Welsh ponies, one miniature Appaloosa and 6 Shetland ponies (adults, mean ± s.d. weight 310 ± 172 kg) suffering from acute or chronic laminitis. EMG measurements including firing frequency (F) and IPA parameters Turns/Second (T), Amplitude/Turn (M) and Ratio M/T (R) were performed. ANOVA was used to analyse data. P values of P<0.05 were considered significant., Results: Mean ± s.d. F of Group 0 and Group 1 was 53 ± 11 and 72 ± 21 Hz, mean ± s.d. T was 112 ± 57 and 106 ± 42, mean ± s.d. M was 284 ± 51 and 254 ± 38 μV and mean ± s.d. R was 0.39 ± 0.17 and 0.42 ± 0.16%, respectively. The firing frequency of Group 1 was significantly higher compared to Group 0 (P = 0.02), whereas other differences were not significant., Conclusions: In human medicine, an increased firing frequency is a characteristic of increased muscle force [1,2]. Thus, the increased firing frequency of the DDFM in case of laminitis suggests an elevated muscle force. As all parameters show a high variance, a repeated study including a larger test group is advised. Ethical animal research: Data collection from controls was approved by the Animal Welfare Committee of Utrecht University, approval number 2008.III.07.061 and 2013.III.01.012. Clinical cases were privately owned and written owner consent was obtained., Source of Funding: None. Competing interests: None declared., (© 2015 The Author(s). Equine Veterinary Journal © 2015 EVJ Ltd.)

Hypophosphatemia is a common finding in periparturient and anorectic cattle. Although the clinical relevance of hypophosphatemia in cattle is uncertain, it has been empirically associated with persistent recumbency, specifically in periparturient dairy cows. The objective of the present study was to determine if transient dietary phosphorus (P) deprivation over a course of 5 wk, by feeding an approximately 40% P-deficient ration to lactating dairy cows, would result in altered muscle function or muscle P metabolism severe enough to present a risk for animal health and well-being. In addition, we wanted to determine the association between the plasma phosphate concentration ([Pi]) and muscle tissue P content to assess to what extent intracellular P deprivation of muscle cells could be extrapolated from subnormal plasma [Pi]. Ten healthy multiparous, mid-lactating dairy cows received a ration with a P content of 0.18% over a period of 5 wk. Following the P-deprivation phase, the same ration supplemented with P to obtain a dietary P content of 0.43% was fed for 2 wk. Blood and urine samples were collected regularly and muscle biopsies were obtained repeatedly to determine the P content in muscle tissue. Function of skeletal and heart muscles was evaluated by electrocardiography and electromyography conducted repeatedly throughout the study. Feeding the P-deficient ration resulted in the rapid development of marked hypophosphatemia. The lowest plasma [Pi] were measured after 9 d of P depletion and were, on average, 60% below predepletion values. Plasma [Pi] increased thereafter, despite ongoing dietary P depletion. None of the animals developed clinical signs commonly associated with hypophosphatemia or any other health issues. Urine analysis revealed increasing renal calcium, pyridinoline, and hydroxypyridinoline excretion with ongoing P deprivation. Biochemical muscle tissue analysis showed that dietary P depletion and hypophosphatemia were not associated with a decline in muscle tissue P content. Electromyographic examination revealed increased occurrence of pathological spontaneous activity in striated muscles after 2 wk of dietary P depletion in several cows, which could be suggestive of neuromuscular membrane instability. No effect on heart muscle activity was identified electrocardiographically. These results suggest that counter-regulatory mechanisms were sufficient to maintain normal muscle tissue P content during transient and moderate P deprivation. Muscle function was not grossly affected, although the increased occurrence of pathological spontaneous activity suggests that subclinical neuropathy or myopathy, or both, may have occurred with ongoing P deprivation. The results presented here indicate that plasma [Pi] is unsuitable for assessing muscle tissue P content in cattle., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

In equine medicine the use of Botox® is experimental. Dosages are determined from human treatment-protocols and limited numbers of equine studies. Determination of target-muscle volume can be helpful to extrapolate human dosages. The aim of the study was to calculate a formula enabling the estimation of the deep digital flexor muscle (DDFM) volume based on distances between anatomical landmarks. Nineteen cadaveric limbs were collected and distance A (top of olecranon to Os carpi accessorium) and B (circumference of limb) were measured. Converting mathematical formulas, C was calculated: π × (((0.5B)/π)(2)) × A. DDFM volume was determined by water displacement. Linear Regression Analysis was used to analyse data. The line best fitting the observed points was: Ln(volume[ml]) = -1.89 + 0.98 × Ln(value C[cm(3)]). Correlation was highest when natural logarithm was applied to both variables and was 0.97. The calculated formula enables estimating DDFM volume of a living horse. This estimated volume can be useful to apply human Botox® treatment-protocols., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Different head-and-neck positions (HNPs) are discussed in relation to potential welfare issues. To evaluate the effect on welfare, seven Royal Dutch Sport horses were studied in five predetermined HNPs: (1) unrestrained (HNP1); (2) neck raised, bridge of nose around the vertical (HNP2); (3) neck lowered and considerably flexed, bridge of nose pointing towards the chest (HNP4); (4) neck raised and extended, bridge of nose in front of the vertical (HNP5), and (5) neck lowered and flexed, bridge of nose pointing towards the carpus (HNP7). A standardised exercise test (SET) of 34 min consisted of trot, canter and walk. Behaviour was recorded with a pre-defined ethogram and R-R intervals measured using telemetry. Cortisol concentrations were taken at the start, 5 and 30 min after the SET. Behaviour around the SET was scored separately. Conflict behaviours increased significantly during HNP2 when compared with HNP1, HNP4 and HNP7 during the SET, and there was significant negative anticipation before HNP2 and HNP7. The heart rate variability (HRV) frequency domain for HNP2 showed a significantly increased low frequency peak (LFpeak) compared with other HNPs, and there was a decrease in very low frequency (VLF%) compared with HNP1. HNP4 showed a significant increase in LF% and decrease in VLF% compared with HNP1. Saliva cortisol concentrations were significantly increased in HNP2 at 5 and 30 min after exercise. Increased conflict behaviour was mostly observed in HNP2, but there was a raised HRV suggesting a sympathetic shift in HNP2 and HNP4, and increased cortisol concentrations during HNP2 indicated a stress response., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: The impact of cervical pathology on performance is of great importance to the horse industry. Accurate diagnosis of cervical disease with imaging modalities, including computed tomography (CT) and magnetic resonance imaging (MRI), requires thorough appreciation of normal cervical anatomy., Objectives: (1) To describe in detail the anatomy of the equine cervical spine by comparing anatomical sections with corresponding MR and contrast-enhanced CT images in the sagittal, dorsal, and transverse plane. (2) To discuss the ability of MR and contrast-enhanced CT imaging to visualize anatomical structures in the cervical spine., Animals and Methods: Three cervical spines of young adults (3-8 years), collected immediately after humane euthanasia, were used. The spine was stabilized on a frame in a natural flexed position with an angle of 20°. MR and contrast-enhanced CT imaging was performed within six hours after euthanasia. Anatomical sections of 1 cm were made in the sagittal, dorsal, and transverse plane and compared with corresponding CT and MR images. The intervertebral disk thickness, facet joint angle, sagittal dural space diameter and ventromedial facet joint projection were quantified., Results: The anatomic location of clinically important structures including the facet joints, spinal cord, cervical nerve roots and intervertebral disks were reliably identified in the anatomical sections and their corresponding MR images. Contrast-enhanced CT images depicted all osseous borders, whereas MR images were superior for soft tissue structures., Conclusion and Clinical Importance: This study enhances our understanding of normal cervical spine anatomy and the diagnostic usefulness of cervical MRI and contrast-enhanced CT in the horse.

In the treatment of laminitis, reducing deep digital flexor muscle (DDFM) activity might diminish its pull on the distal phalanx, thereby preventing displacement and providing pain relief. Injection of Clostridium botulinum toxin type A into the DDFM of horses is potentially therapeutic. However, the effects of C. botulinum toxin type A on the gait characteristics of sound horses at the walk are not known. The aim of this study was to test if a reduced DDFM activity would lead to (1) alterations of the sagittal range of motion of the metacarpus (SROM) and range of motion of the carpal joint (CROM); (2) changes in the force distribution underneath the hoof (toe vs. heel region: balance index); and (3) changes in the force distribution between the treated and untreated limb (symmetry index). The DDFMs of the left forelimbs of seven sound Royal Dutch Sport Horses were injected with 200 IU C. botulinum toxin type A using electromyography and ultrasound guidance. Measurements using an inertial sensor system and dynamically calibrated pressure plate were performed before and after injections. The SROM and CROM of the treated limb were significantly increased after C. botulinum toxin type A injections. No significant changes were detected in the balance index or in the symmetry index, indicating that no lameness was induced. C. botulinum toxin type A injections into the DDFM of sound horses do not appear to result in substantial gait alterations at the walk., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Therapeutic reduction of the activity of the deep digital flexor (DDF) muscle may play a role in treatment of laminitic horses. Clostridium botulinum toxin type A induces reduced muscle activity and has a spasmolytic effect in horses. In this study, the effectiveness of 200 IU C. botulinum toxin type A on reduction of DDF muscle activity was measured in seven healthy, sound, adult Royal Dutch sport horses. C. botulinum toxin type A was injected using ultrasound and electromyographic (EMG) guidance. The effectiveness was assessed by interference pattern analysis (IPA) and motor unit action potential (MUAP) analysis. All needle EMG MUAP variables, along with IPA amplitude/turn and turns/s, were significantly reduced after C. botulinum toxin type A injections. The strongest effect occurred within the first 3 days after injection. The reduced muscle induced by C. botulinum toxin type A may have benefits in the treatment of horses with laminitis., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Empirically, equine distal limb lameness is often linked to hoof imbalance. To objectively quantify dynamic toe-heel and medio-lateral hoof balance of the vertical ground reaction force in sound sport horses, seven Royal Dutch Sport Horses were led at the walk and trot over a dynamically calibrated pressure plate. Forelimb hoof prints were divided into a toe and heel region and a medial and lateral zone. Toe-heel and medio-lateral hoof balance of the vertical ground reaction force were calculated throughout the stance. Toe-heel balance was highly symmetrical between contralateral limbs at both gaits. At the walk, medio-lateral balance of both forelimbs presented higher loading in the lateral part of the hoof throughout the stance. However, at the trot, left medio-lateral balance presented higher loading of the medial part of the hoof at impact, whereas the right limb showed higher loading of the lateral part of the hoof in all horses, and both limbs presented increased lateral loading at the end of the stance. This study provides objective data for toe-heel and medio-lateral hoof balance in sound sport horses., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Training horses improves athletic capabilities by inducing skeletal muscle-specific and systemic adaptations. However, rest is required to recover from exercise or else overtraining may occur and affect performance and welfare. Biomarkers would be useful to identify early chronic overtraining in animals. The objective of the current study was to investigate skeletal muscle gene expression patterns and underlying biological mechanisms related to training of different intensities and detraining. Untrained 20 month-old Standardbred geldings were exercised at varying intensities (endurance and sprint) followed by detraining (n=5 per phase). The results indicated that training mainly affected skeletal muscle-specific protein metabolism and increased CO2 export from the tissues. Intensive training increased energy metabolism and affected heart and adipose tissues, while having an adverse effect on stress, apoptosis and immune capacity. The intensity of the training could be related to decreased expression of extra cellular matrix proteins (ECM), cell-cell contacts and intracellular signalling pathways. During detraining, most mechanisms were reversed, but heart tissue-related changes and increased expression of skeletal muscle-specific proteins were still evident. The study suggested that changes to ECM expression and cell-cell contact mechanisms may be long-lasting and related to multifactorial aspects of training and detraining. These biomarkers may be useful to identify horses in the early stages of chronic overloading or early overtraining., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Background: Short-duration electroencephalography (EEG) recordings in horses are helpful in diagnosing intracranial disorders. Potentially, long-duration ambulatory EEG (AEEG) recordings in horses will enhance the chance of detecting abnormal brain activity independent of the presence of an insult., Objective: The objective of this study was to test if AEEG recordings in unsedated horses can be acquired and benefit diagnosing abnormal brain activity., Animals and Methods: Recordings were taken from 8 adult control horses and 10 patients suspected of intracranial abnormalities. Self-adhesive electrodes and the 'Porti-5' recording system were used. Filter settings were 0.5 Hz high pass and 35 Hz low pass. The records were analysed offline at a 50-200 μV/division and 10 seconds/division scale. Abnormal activity was defined as a spike or sharp wave, a period of generalised slow wave rhythmical activity or a generalised fast rhythmical discharge. The recording time ranged from 5 to 49 hours., Results: In the control group, one horse showed pathological activity. In the patient group, six out of nine horses showed abnormal activity during the recordings. Magnetic resonance imaging confirmed the presence of an intracranial mass in one patient. Long-term recordings of high quality can be obtained in unsedated horses by allowing daily activity using AEEG, resulting in a reasonable chance of recording (inter)ictal abnormal brain activity indicating epileptic or seizure-like activity in the absence of clinical signs or seizures., Conclusions: It is concluded that abnormal behaviour can be expressed intermittently, and with the availability of AEEG a useful tool is added to the diagnostic scenario for horses.

Objective: To determine the influence of intensified training and subsequent reduced training on glucose metabolism rate and peripheral insulin sensitivity in horses and identify potential markers indicative of early overtraining., Animals: 12 Standardbred geldings., Procedures: Horses underwent 4 phases of treadmill-based training. In phase 1, horses were habituated to the treadmill. In phase 2, endurance training was alternated with high-intensity exercise training. In phase 3, horses were divided into control and intensified training groups. In the intensified training group, training intensity, duration, and frequency were further increased via a protocol to induce overtraining; in the control group, these factors remained unaltered. In phase 4, training intensity was reduced. Standardized exercise tests were performed after each phase and hyperinsulinemic euglycemic clamp (HEC) tests were performed after phases 2, 3, and 4., Results: 10 of 12 horses completed the study. Dissociation between mean glucose metabolism rate and mean glucose metabolism rate-to-plasma insulin concentration ratio (M:I) was evident in the intensified training group during steady state of HEC testing after phases 3 and 4. After phase 4, mean glucose metabolism rate was significantly decreased (from 31.1 ± 6.8 μmol/kg/min to 18.1 ± 3.4 μmol/kg/min), as was M:I (from 1.05 ± 0.31 to 0.62 ± 0.17) during steady state in the intensified training group, compared with phase 3 values for the same horses., Conclusions and Clinical Relevance: Dissociation between the glucose metabolism rate and M:I in horses that underwent intensified training may reflect non-insulin-dependent increases in glucose metabolism.

A 6-year-old Friesian stallion was examined because of signs of exercise intolerance, stiff gait and symmetrical hind weakness, and increased serum liver enzymes. On presentation, the horse showed muscle atrophy of the hindquarters. Neurological investigation showed no abnormalities. Laboratory findings revealed a prolonged prothrombin time and increased levels of alkaline phosphatase (AF), aspartate aminotransferase (ASAT), gamma-glutamyl-transferase (GGT), lactate dehydrogenase (LDH), and bile acids. Histological evaluation of the liver revealed severe cirrhosis and intracytoplasmic greyish brown granules in almost all hepatocytes, sinusoidal Kuppfer cells, and macrophages. These granules stained strongly for copper. Treatment to slow hepatic fibrosis was advised and included oral prednisolone administration for at least 1 month. A diet to support liver function was formulated by a nutritional specialist, and vitamin E was advised as dietary supplement to support neuromuscular function. Soon after diagnosis, the animal showed signs of intravascular haemolysis, with the presence of Heinz bodies in peripheral blood smears, and haemoglobinuria. On the basis of this haemolytic crisis and the poor prognosis of the chronic hepatic disease, the horse was euthanized at the owners' request. Although we could not establish the cause of the hepatic copper accumulation, this case report highlights that excessive copper in the liver should be considered in the differential diagnosis of hepatic cirrhosis and Heinz body anaemia in the horse.

A 7-month-old New Forest foal presented for episodes of recumbency and stiffness with myotonic discharges on electromyography. The observed phenotype resembled congenital myotonia caused by CLCN1 mutations in goats and humans. Mutation of the CLCN1 gene was considered as possible cause and mutation analysis was performed. The affected foal was homozygous for a missense mutation (c.1775A>C, p.D592A) located in a well conserved domain of the CLCN1 gene. The mutation showed a recessive mode of inheritance within the reported pony family. Therefore, this CLCN1 polymorphism is considered to be a possible cause of congenital myotonia., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Objective: To evaluate the effect of various head and neck positions on intrathoracic pressure and arterial oxygenation during exercise in horses., Animals: 7 healthy Dutch Warmblood riding horses., Procedures: The horses were evaluated with the head and neck in the following predefined positions: position 1, free and unrestrained; position 2, neck raised with the bridge of the nose aligned vertically; position 4, neck lowered and extremely flexed with the nose pointing toward the pectoral muscles; position 5, neck raised and extended with the bridge of the nose in front of a vertical line perpendicular to the ground surface; and position 7, neck lowered and flexed with the nose pointing towards the carpus. The standard exercise protocol consisted of trotting for 10 minutes, cantering for 4 minutes, trotting again for 5 minutes, and walking for 5 minutes. An esophageal balloon catheter was used to indirectly measure intrathoracic pressure. Arterial blood samples were obtained for measurement of Pao(2), Paco(2), and arterial oxygen saturation., Results: Compared with when horses were in the unrestrained position, inspiratory intrathoracic pressure became more negative during the first trot (all positions), canter and second trot (position 4), and walk (positions 4 and 5). Compared with when horses were in position 1, intrathoracic pressure difference increased in positions 4, 2, 7, and 5; Pao(2) increased in position 5; and arterial oxygen saturation increased in positions 4 and 7., Conclusions and Clinical Relevance: Position 4 was particularly influential on intrathoracic pressure during exercise in horses. The effects detected may have been caused by a dynamic upper airway obstruction and may be more profound in horses with upper airway disease.

Reasons for Performing Study: To compare the pharmacokinetics of the fourth generation cephalosporin, cefquinome, in neonatal foals, 6-week-old foals and mature New Forest ponies in order to recommend appropriate dosage regimens for use of this drug., Methods: Cefquinome was administered i.v. at 1 mg/kg bwt twice a day (q. 12 h), 1 mg/kg bwt 3 times a day (q. 8 h) or 4.5 mg/kg bwt q. 12 h to each age group (n = 6). Plasma cefquinome concentrations were analysed using high-performance liquid chromatography combined with electrospray tandem mass spectrometry., Results: Both foal age groups had comparable pharmacokinetic data except for the volume of distribution at a steady-state (Vss), total body clearance (ClB) and mean residence time (MRT). Both ClB and MRT decreased as the age of the foals increased. Values of area under the curve increased, in a dose dependent manner, with significant increases for all age groups following administration of 4.5 mg/kg bwt q. 12 h. Total body clearance did not have comparable dose dependency., Conclusions: Cefquinome can be given at a dose of 1 mg/kg bwt q. 12 h for the treatment of infections caused by susceptible pathogens with MIC < 0.125 microg/ml. A higher dose of 4.5 mg/kg bwt q. 12 h is recommended for the treatment of bacterial pathogens with minimal inhibitory concentration (MIC) 0.125-0.5 microg/ml, Potential Relevance: Commonly used dosing regimens should be critically evaluated in neonatal foals due to the higher volume of distribution of less lipophilic drugs in this age group.

The objective of this study is to assess the influence of acute exercise, training and intensified training on the plasma amino acid profile. In a 32-week longitudinal study using 10 Standardbred horses, training was divided into four phases, including a phase of intensified training for five horses. At the end of each phase, a standardized exercise test, SET, was performed. Plasma amino acid concentrations before and after each SET were measured. Training significantly reduced mean plasma aspartic acid concentration, whereas exercise significantly increased the plasma concentrations of alanine, taurine, methionine, leucine, tyrosine and phenylalanine and reduced the plasma concentrations of glycine, ornithine, glutamine, citrulline and serine. Normally and intensified trained horses differed not significantly. It is concluded that amino acids should not be regarded as limiting training performance in Standardbreds except for aspartic acid which is the most likely candidate for supplementation., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Hypothesis/objectives: Defining normal Growth Hormone (GH) secretory dynamics in the horse is necessary to understand altered GH dynamics related to issues like welfare and disease., Animals and Methods: Twelve healthy yearlings and two mature Standardbreds were used to quantify GH secretion. Endogenous GH half-life was determined after administration of 1.0 µg/kg BW GH releasing hormone (GHRH). Exogenous GH half-life was determined after administration of 20 µg/kg BW recombinant equine GH (reGH) with and without suppression of endogenous GH secretion by somatostatin infusion (50 µg/m(2)/h). Pulse detection algorithm (Cluster) as well as deconvolution analysis was used to quantify GH secretory dynamics based on GH concentration-time series sampled every 5 min from 22:00 till 06:00 h. In addition, reproducibility, impact of sampling frequency and influence of altering initial GH half-life on parameter estimates were studied., Results: Mean endogenous GH half-life of 17.7 ± 4.4 (SD) min and mean exogenous half-life of 26.0 ± 2.9 min were found. The mean number of GH secretion peaks in 8 h was 12 ± 3.2. Ninety-nine percent of the total amount of GH secreted occurred in pulses, basal secretion was 0.012 ± 0.014 µg/L/min and half-life was 8.9 ± 2.6 min. Compared with a 5-min sampling frequency, 20- and 30-min sampling underestimated the number of secretory events by 45% and 100%, respectively., Conclusions: The deconvolution model used was valid to GH time series in Standardbreds. As in man, the equine pituitary gland secretes GH in volleys consisting of multiple secretory bursts, without measurable intervening tonic secretion. The required GH sampling frequency for the horse should be around 3 min., Clinical Relevance: Defining normal GH secretory dynamics in the horse will make it possible to detect alterations in the GH axis due to pathophysiologic mechanisms as well as abuse of reGH.

Background: Reference values for quantitative electromyography (QEMG) in neck muscles of Royal Dutch Sport horses are lacking., Objective: Determine normative data on quantitative motor unit action potential (QMUP) analysis of serratus ventralis cervicis (SV) and brachiocephalicus (BC) muscle., Animals: Seven adult normal horses (mean age 9.5 standard deviation [SD] ± 2.3 years, mean height 1.64 SD ± 4.5 cm, and mean rectal temperature 37.6 SD ± 0.3°C)., Methods: An observational study on QMUP analysis in 6 segments of each muscle was performed with commercial electromyography equipment. Measurements were made according to formerly published methods. Natural logarithm transformed data were tested with ANOVA and posthoc testing according to Bonferroni., Results: Mean duration, amplitude, phases, turns, area, and size index (SI) did not differ significantly among the 6 segments in each muscle. Mean amplitude, number of phases, and SI were significantly (P < .002) higher in SV than BC, 520 versus 448 μV, 3.0 versus 2.8 μV, and 0.48 versus 0.30 μV, respectively. In SV 95% confidence intervals (CI) for amplitude, duration, number of phases, turns, polyphasia area, and SI were 488-551 μV, 4.3-4.6 ms, 2.9-3.0, 2.4-2.6, 7-12%, 382-448, and 0.26-0.70, respectively; in BC this was 412-483 μV, 4.3-4.7 ms, 2.7-2.8, 2.4-2.6, 4-7%, 393-469, and 0.27-0.34, respectively. Maximal voluntary activity expressed by turns/second did not differ significantly between SV and BC with a 95% CI of 132-173 and 137-198, respectively., Conclusions and Clinical Importance: The establishment of normative data makes objective QEMG of paraspinal muscles in horses suspected of cervical neurogenic disorders possible. Differences between muscles should be taken into account., (Copyright © 2011 by the American College of Veterinary Internal Medicine.)

Reasons for Performing Study: In dressage, the head and neck position has become an issue of concern as certain extreme positions may imply a welfare risk for the horse. In man, extension and flexion of the cervical spine cause a decrease and increase in intervertebral foramina dimensions, respectively. However, in horses, the influence of flexion and extension on foramina dimensions and its possible interference with peripheral nerve functioning remains unknown., Objectives: To determine the effect of ex vivo flexion and extension on intervertebral foramina dimensions in the equine cervical spine., Methods: Computed tomography was performed on 6 cadaver cervical spines from adult Warmblood horses subjected to euthanasia for reasons unrelated to cervical spine abnormalities, in a neutral position, in 20 and 40° extension, and in 20 and 40° flexion. Multiplanar reconstructions were made to obtain transverse images perpendicular to the long axis of each pair of intervertebral foramina from C2-T1. Intervertebral foramina dimensions were measured in the 5 positions., Results: Compared to the neutral position, 40° extension caused a decrease in foramina dimensions at segments C4-C5, C5-C6, C6-C7 (P < 0.001) and C7-T1 (P < 0.002); 20° extension caused a decrease in foramina dimensions at segments C5-C6 (P < 0.02), C6-C7 (P < 0.001) and C7-T1 (P < 0.01); 20° flexion caused an increase in foramen length at segment C6-C7 (P < 0.01)., Conclusions: Ex vivo extension of the cervical spine causes a decrease in intervertebral foramina dimensions at segments C4-T1, similar to that found in man., Potential Relevance: In vivo extension of the cervical spine could possibly interfere with peripheral nerve functioning at segments C4-T1. This effect may be even more profound in patients with a reduced intervertebral foramina space, for example in the presence of facet joint arthrosis., (© 2010 EVJ Ltd.)

Reasons for Performing Study: There has been growing interest in training techniques with respect to the head and neck position (HNP) of the equine athlete. Little is known about the influence of HNP on neuromuscular transmission in neck muscles., Objective: To test the hypothesis that different HNPs have effect on single fibre (SF), quantitative electromyographic (QEMG) examination and muscle enzyme activity directly after moderate exercise., Methods: Seven Warmblood horses were studied using a standard exercise protocol in 5 HNPs: HNP1: unrestrained; HNP2: neck raised; bridge of nose around the vertical; HNP4: neck lowered and considerably flexed, bridge of nose pointing towards the chest; HNP5: neck raised and considerably extended; bridge of nose in front of the vertical; HNP7: neck lowered and flexed; bridge of nose pointing towards the carpus. Mean consecutive difference (MCD) of single muscle fibre potentials and motor unit action potential (MUP) variables (amplitude, duration, area, turns and phases) were recorded in each fixed position directly after exercise at rest using commercial EMG equipment. Muscle enzyme activity was measured before and 4, 6 and 24 h after exercise., Results: Mean consecutive difference in all HNPs was higher than in HNP1 (22 µs, P < 0.001) of which HNP4 was highest with 39 µs compared to 30 µs in HNP2 (P = 0.04); MCD in HNP 5,7 was with 25 µs lower than in HNP 2 and 4 (P < 0.001). Odds ratio for MCD suggestive for conduction delay or block was 13.6 in HNP4 compared to HNP1 (P < 0.001). Motion unit action potential variables followed the same pattern as MCD. Lactate dehydrogenase (LDH) activity increased in HNP4 at 4 h (P = 0.014), 6 h (P = 0.017) and 24 h (P = 0.038) post exercise and in HNP5 and HNP7 at 4 h (P = 0.037; 0.029)., Conclusions and Clinical Relevance: HNP4 in particular leads to a higher rise in LDH activity, MCD and MUP variables, indicating that HNPs have effect on variables characterising neuromuscular functionality., (© 2010 EVJ Ltd.)

Reasons for Performing Study: Head and neck positions (HNP) in sport horses are under debate in the equine community, as they could interfere with equine welfare. HNPs have not been quantified objectively and no information is available on their head and neck loading., Objectives: To quantify in vivo HNPs in sport horses and develop o a model to estimate loading on the cervical vertebrae in these positions., Methods: Videos were taken of 7 Warmbloods at walk on a straight line in 5 positions, representing all HNPs during Warmblood training and competition. Markers were glued at 5 anatomical landmarks. Two-dimensional angles and distances were determined from video frames for the 5 HNPs and statistically compared (P < 0.05). A new simulation model was developed to estimate nuchal ligament cervical loading at these HNPs., Results: The mean angles were significantly different between the 5 HNPs for the line between C1 and T6 with the horizontal and for the line connecting the facial crest (CF) and C1 with the vertical, while the vertical distance from CF to the lateral styloid process of the radius (PS) was significantly different between all 5 positions (P < 0.05). The estimated nuchal ligament loading appeared to be largest at the origin of C2 for all HNPs, except for the 'hyperextended' HNP5; the 'hyperflexed' HNP4 showed the largest loading values on the nuchal ligament origins at all locations., Conclusions: HNPs can be accurately quantified in the sagittal plane from angles and distances based on standard anatomical landmarks and home-video captured images. Nuchal ligament loading showed the largest estimated values at its origin on C2 in hyperflexion (HNP4)., Potential Relevance: Modelling opens further perspectives to eventually estimate loading for individual horses and thus ergonomically optimise their HNP, which may improve the welfare of the sport horse during training and competition., (© 2010 EVJ Ltd.)

Reasons for Performing Study: Gastric ulceration can be caused by different pathophysiological mechanisms including dietary factors, psychological stress and exercise. Overtraining is a medical syndrome in performance horses associated with altered hormone levels, altered feed intake, altered behaviour and decreased performance. These components might lead to a higher incidence of gastric ulceration in overtrained horses., Objectives: To investigate whether the incidence of gastric ulceration is increased in overtrained compared to control horses., Methods: A longitudinal training study with twelve 1.5 years old Standardbred horses was performed on a treadmill for a total of 32 weeks. Training was divided into 4 periods: (1) acclimatisation (2) training (3) intensified training, and (4) detraining. In period 3, the horses were randomly divided into 2 groups: control (C) and intensified trained group (IT). At the end of each period, gastroscopy was performed in conscious horses after withholding feed for 12 h and water for 6 h using a 3.5 m video gastroendoscope. Lesion scores were assigned to areas of the stomach and graded 1-4. Logistic regression was used for statistical calculations., Results: Evaluation of the stomach revealed only minor changes (grades 1 or 2) on each occasion. There were no significant differences in gastric lesion scores between groups or periods. Most lesions (70%) were found around the minor curvature. After detraining no lesions (0%) were found in contrast to periods 1 (40%, P = 0.056), 2 (30%) and 3 (30%)., Conclusions: Experimentally-induced overtraining does not increase the incidence of gastric ulceration in normally fed Standardbred horses and detraining appears to reduce gastric ulceration., (© 2010 EVJ Ltd.)

This case-series describes fourteen horses suspected of equine acquired multiple acyl-CoA dehydrogenase deficiency (MADD) also known as atypical myopathy of which seven cases were confirmed biochemically with all horses having had access to leaves of the Maple tree (Acer pseudoplatanus) covered with European tar spot (Rhytisma acerinum). Assessment of organic acids, glycine conjugates, and acylcarnitines in urine was regarded as gold standard in the biochemical diagnosis of equine acquired multiple acyl-CoA dehydrogenase deficiency., (Copyright © 2010 Elsevier Inc. All rights reserved.)

The major aim of the present study was to investigate the proteome of standardbred horses at different stages of training and intensified training. We searched for biomarkers using small skeletal muscle biopsies of live animals. 2D gel electrophoresis and mass spectrometry were successfully applied to investigate training-induced differential expression of equine muscle biopsy proteins. Despite the poor resolution of the equine genome and proteome, we were able to identify the proteins of 20 differential spots representing 16 different proteins. Evaluation of those proteins complies with adaptation of the skeletal muscle after normal training involving structural changes towards a higher oxidative capacity, an increased capacity to take up long-chain fatty acids, and to store energy in the form of glycogen. Intensified training leads to additional changed spots. Alpha-1-antitrypsin was found increased after intensified training but not after normal training. This protein may thus be considered as a marker for overtraining in horses and also linked to overtraining in human athletes., (Copyright 2009 Elsevier Inc. All rights reserved.)

The influence of intensified and reduced training on nocturnal growth hormone (GH) secretion and elimination dynamics was studied in young (1.5 yr) Standardbred geldings to detect potential markers indicative for early overtraining. Ten horses trained on a treadmill for 32 wk in age-, breed-, and gender-matched fixed pairs. Training was divided into four phases (4, 18, 6, and 4 wk, respectively): 1) habituation to high-speed treadmill trotting, 2) normal training, in which speed and duration of training sessions were gradually increased, 3) in this phase, the horses were divided into 2 groups: control (C) and intensified trained (IT) group. In IT, training intensity, duration, and frequency were further increased, whereas in control these remained unaltered, and 4) reduced training (RT). At the end of phases 2, 3, and 4, blood was sampled overnight every 5 min for 8 h for assessment of GH secretory dynamics using pulse detection, deconvolution analysis, and approximate entropy (ApEn). Intensified training induced overtraining (performance decreased by 19% compared with C), which was associated with an increase in concentration peaks number (3.6 vs. 2.0, respectively), a smaller peak secretion pattern with a prolonged half-life (15.2 vs. 7.3 min, respectively), and an increased ApEn (0.89 vs. 0.49, respectively). RT did not lead to full recovery for the overtrained horses. The increased irregularity of nocturnal GH pulsatility pattern is indicative of a loss of coordinated control of GH regulation. Longer phases of somatostatin withdrawal are hypothesized to be the underlying mechanism for the observed changes in GH pulsatility pattern.

Objective: To investigate the effects of acute exercise and long-term training on Na(+),K(+)-ATPase content, mRNA isoforms, and protein concentration in equine muscle., Animals: 6 Standardbreds., Procedures: Horses performed a bout of exercise on a treadmill before and after 18 weeks of combined interval and endurance training. Muscle biopsy specimens were obtained from vastus lateralis muscle (VLM) and pectoralis descendens muscle (PDM) before and after exercise. The Na(+),K(+)-ATPase content, mRNA isoforms, and protein concentrations were determined by use of [(3)H]ouabain binding, real-time PCR assay, and western blotting, respectively., Results: 6 Na(+),K(+)-ATPase mRNA isoforms were present in equine muscle, but only A2 and B1 proteins were detected. Exercise before training resulted in increases of mRNA isoforms A1, A2, A3, and B2 in VLM and A1 and B3 in PDM. Training increased resting values for mRNA isoforms A3 and B1 in VLM and B3 in PDM. The Na(+),K(+)-ATPase, [(3)H]ouabain binding, and proteins of mRNA A2 and B1 increased in VLM, whereas in PDM, only A2 protein increased as a result of training. After training, effects of strenuous exercise on mRNA expression were no longer detectable., Conclusions and Clinical Relevance: Equine muscle contained all Na(+),K(+)-ATPase mRNA isoforms, but only A2 and B1 proteins could be detected. Expression of these isoforms changed as a result of strenuous exercise and long-term training, representing an adaptive response. Determination of Na(+),K(+)-ATPase gene expression may be relevant for understanding alterations in excitability during neuromuscular diseases.

Reasons for Performing the Study: Therapeutic options for stringhalt in horses are limited, whereas medical experiences with botulinum toxin type A (Botox) have been positive. To evaluate its effectiveness in horses, surface electromyography (sEMG) signals before and after injection need to be quantified., Hypothesis: Treatment of healthy ponies and cases with Botox should reduce muscle activity in injected muscles and reduce spastic movements without adverse side effects., Methods: Unilaterally, the extensor digitorum longus, extensor digitorum lateralis and lateral vastus muscles of 6 healthy mature Shetland ponies and 2 talented Dutch Warmblood dressage horses with stringhalt were injected (maximum of 400 iu per pony and 700 iu per case; 100 iu in 5 ml NaCl divided into 5 injections) with Botox under needle EMG guidance. Surface EMG data were evaluated using customised software, and in the individuals gait was analysed using Proreflex. Statistical analysis was performed using mixed models and independent sample t test (P < 0.05)., Results: Surface EMG signals were quantified using customised software. The area under the curve (integrated EMG) in time was used as variable. It became significantly reduced in injected muscles after injection of Botox in normal ponies (P < 0.05). This effect was present from Day 1 until Day 84 after injection. In the 2 cases, after injection of 3 muscles, the integrated EMG in time became significantly reduced in all 3 muscles. Kinematic measurements confirmed reduction of frequency and amplitude of hyperflexing or hyperabducting strides of the affected hindlimbs. The duration of effect was also seen in the cases until around 12 weeks after injection., Conclusions and Potential Relevance: After EMG guided injections of Botox, sEMG signals recorded from injected muscle were reduced, which proves this to be a useful tool in statistically evaluating a treatment effect. The positive results of this pilot study encourage further research with a larger group of clinical cases.

Too intensive training may lead to overreaching or overtraining. To study whether quantitative needle electromyography (QEMG) is more sensitive to detect training (mal)adaptation than muscle enzyme activities, 12 standardbred geldings trained for 32 wk in age-, breed-, and sex-matched fixed pairs. After a habituation and normal training (NT) phase (phases 1 and 2, 4 and 18 wk, respectively), with increasing intensity and duration and frequency of training sessions, an intensified training (IT) group (phase 3, 6 wk) and a control group (which continued training as in the last week of phase 2) were formed. Thereafter, all horses entered a reduced training phase (phase 4, 4 wk). One hour before a standardized exercise test (SET; treadmill), QEMG analysis and biochemical enzyme activity were performed in muscle or in biopsies from vastus lateralis and pectoralis descendens muscle in order to identify causes of changes in exercise performance and eventual (mal)adaptation in skeletal muscle. NT resulted in a significant adaptation of QEMG parameters, whereas in muscle biopsies hexokinase activity was significantly decreased. Compared with NT controls, IT induced a stronger adaptation (e.g., higher amplitude, shorter duration, and fewer turns) in QEMG variables resembling potentially synchronization of individual motor unit fiber action potentials. Despite a 19% decrease in performance of the SET after IT, enzyme activities of 3-hydroxyacyl dehydrogenase and citrate synthase displayed similar increases in control and IT animals. We conclude that 1) QEMG analysis is a more sensitive tool to monitor training adaptation than muscle enzyme activities but does not discriminate between overreaching and normal training adaptations at this training level and 2) the decreased performance as noted in this study after IT originates most likely from a central (brain) rather than peripheral level.

Objective: To evaluate alterations in skeletal muscle carnitine metabolism during exercise and training by measuring changes in plasma acylcarnitine concentrations in Standardbreds., Animals: 10 Standardbred geldings with a mean +/- SD age of 20 +/- 2 months and weight of 384 +/- 42 kg., Procedures: In a 32-week longitudinal study, training on a treadmill was divided into 4 phases as follows: phase 1, acclimatization for 4 weeks; phase 2, 18 weeks with alternating endurance and high-intensity exercise training; phase 3, increased training volume and intensity for another 6 weeks; and phase 4, deconditioning for 4 weeks. In phase 3, horses were randomly assigned to 2 groups as follows: control horses (which continued training at the same level as in phase 2) and high-intensity exercise trained horses. At the end of each phase, a standardized exercise test (SET) was performed. Plasma acylcarnitine, fatty acids, and lactic acid and serum beta-hydroxybutyric acid (BHBA) concentrations were assessed before and at different time points after each SET., Results: Plasma lactic acid, total nonesterified fatty acids, 3-hydroxyisobutyric acid, and acetylcarnitine (C2-carnitine) concentrations significantly increased during SETs, whereas serum BHBA, plasma propionylcarnitine (C3-carnitine), and plasma butyryl- and isobutyrylcarnitine (C4-carnitine) concentrations decreased significantly, compared with those before SETs., Conclusions and Clinical Relevance: Our findings indicated that the plasma acylcarnitine profile in horses likely reflects skeletal muscle carnitine metabolism following exercise, thereby providing a possible practical method to investigate potential disorders in carnitine metabolism in horses with myopathy.

The aim of the current study was to assess lipid metabolism in horses with atypical myopathy. Urine samples from 10 cases were subjected to analysis of organic acids, glycine conjugates, and acylcarnitines revealing increased mean excretion of lactic acid, ethylmalonic acid, 2-methylsuccinic acid, butyrylglycine, (iso)valerylglycine, hexanoylglycine, free carnitine, C2-, C3-, C4-, C5-, C6-, C8-, C8:1-, C10:1-, and C10:2-carnitine as compared with 15 control horses (12 healthy and three with acute myopathy due to other causes). Analysis of plasma revealed similar results for these predominantly short-chain acylcarnitines. Furthermore, measurement of dehydrogenase activities in lateral vastus muscle from one horse with atypical myopathy indeed showed deficiencies of short-chain acyl-CoA dehydrogenase (0.66 as compared with 2.27 and 2.48 in two controls), medium-chain acyl-CoA dehydrogenase (0.36 as compared with 4.31 and 4.82 in two controls) and isovaleryl-CoA dehydrogenase (0.74 as compared with 1.43 and 1.61 nmol min(-1) mg(-1) in two controls). A deficiency of several mitochondrial dehydrogenases that utilize flavin adenine dinucleotide as cofactor including the acyl-CoA dehydrogenases of fatty acid beta-oxidation, and enzymes that degrade the CoA-esters of glutaric acid, isovaleric acid, 2-methylbutyric acid, isobutyric acid, and sarcosine was suspected in 10 out of 10 cases as the possible etiology for a highly fatal and prevalent toxic equine muscle disease similar to the combined metabolic derangements seen in human multiple acyl-CoA dehydrogenase deficiency also known as glutaric acidemia type II.

Overtraining is an imbalance between training and recovery leading to symptoms associated with a neuroendocrine dysbalance called the overtraining syndrome, a disease characterized by behavioral, emotional and physical symptoms similar with depression. Although the prevalence of overtraining is high in human and equine athletes, at present no sensitive and specific test is available to prevent or diagnose overtraining. Nowadays, it is believed that combination of different (hormonal) parameters appear to be the best indicators of overtraining. Therefore, this review provides a summary of previous literature examining the response of the hypothalamic-pituitary-adrenal (HPA) axis and the growth hormone-insulin-like growth factor-I (GH-IGF-I) axis to acute and chronic exercise as well as overtraining in humans and horses. The exercise induced hormonal responses seem to be equal for the equine as well as the human athlete, which makes comparisons possible. Repeated bouts of exercise are suggested to provide a way to detect subtle changes in hormonal responses in the individual athlete, which may make them an important tool in detecting early overtraining. This should be combined with corticotropin releasing hormone (CRH) stimulation tests and basal ACTH and GH pulsatility determination. Further research is needed to establish the correct training intensity and rest period for the exercise test in equines.

Two horses (a 7-year-old Groninger warmblood gelding and a six-month-old Trakehner mare) with pathologically confirmed rhabdomyolysis were diagnosed as suffering from multiple acyl-CoA dehydrogenase deficiency (MADD). This disorder has not been recognised in animals before. Clinical signs of both horses were a stiff, insecure gait, myoglobinuria, and finally recumbency. Urine, plasma, and muscle tissues were investigated. Analysis of plasma showed hyperglycemia, lactic acidemia, increased activity of muscle enzymes (ASAT, LDH, CK), and impaired kidney function (increased urea and creatinine). The most remarkable findings of organic acids in urine of both horses were increased lactic acid, ethylmalonic acid (EMA), 2-methylsuccinic acid, butyrylglycine (iso)valerylglycine, and hexanoylglycine. EMA was also increased in plasma of both animals. Furthermore, the profile of acylcarnitines in plasma from both animals showed a substantial elevation of C4-, C5-, C6-, C8-, and C5-DC-carnitine. Concentrations of acylcarnitines in urine of both animals revealed increased excretions of C2-, C3-, C4-, C5-, C6-, C5-OH-, C8-, C10:1-, C10-, and C5-DC-carnitine. In addition, concentrations of free carnitine were also increased. Quantitative biochemical measurement of enzyme activities in muscle tissue showed deficiencies of short-chain acyl-CoA dehydrogenase (SCAD), medium-chain acyl-CoA dehydrogenase (MCAD), and isovaleryl-CoA dehydrogenase (IVD) also indicating MADD. Histology revealed extensive rhabdomyolysis with microvesicular lipidosis predominantly in type 1 muscle fibers and mitochondrial damage. However, the ETF and ETF-QO activities were within normal limits indicating the metabolic disorder to be acquired rather than inherited. To our knowledge, these are the first cases of biochemical MADD reported in equine medicine.

This review gives an overview of the presently known human and equine metabolic myopathies with emphasis on the diagnostic approach. Metabolic myopathies are muscle disorders caused by a biochemical defect of the skeletal muscle energy system, which results in inefficient muscle performance. Myopathies can arise in different levels of the metabolic system. In this review the metabolic myopathies are categorized in disorders of the carbohydrate metabolism, lipid metabolism, mitochondrial myopathies (other than those described in lipid metabolism), disorders of purine metabolism, primary disorders involving ion channels and electrolyte flux and secondary or acquired metabolic myopathies.