Evaluation of the risk of motor neuron disease in horses fed a diet low in vitamin E and high in copper and iron

OBJECTIVE: To determine whether equine motor neuron disease (EMND) could be induced in adult horses fed a diet low in vitamin E and high in copper and iron. ANIMALS: 59 healthy adult horses. PROCEDURE: Horses in the experimental group (n = 8) were confined to a dirt lot and fed a concentrate low in vitamin E and high in iron and copper in addition to free-choice grass hay that had been stored for 1 year. Control horses (n = 51) were fed a concentrate containing National Research Council-recommended amounts of copper, iron, and vitamin E. The hay fed to control horses was the same as that fed to experimental horses, but it had not been subjected to prolonged storage. Control horses had seasonal access to pasture, whereas experimental horses had no access to pasture. Horses that developed clinical signs of EMND were euthanatized along with an age-matched control horse to determine differences in hepatic concentrations of vitamin E, vitamin A, copper, iron, and selenium. RESULTS: 4 experimental horses developed clinical signs of EMND. Plasma concentrations of vitamin E decreased in all 8 experimental horses. There were no significant changes in plasma concentrations of vitamin A, selenium, and copper or serum concentrations of ferritin. There were no significant differences in those analytes between experimental horses with EMND and experimental horses that did not develop EMND. No control horses developed EMND. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that lack of access to pasture, dietary deficiency of vitamin E, or excessive dietary copper are likely risk factors for EMND.     

Differential expression of TAR DNA-binding protein (TDP-43) in the central nervous system of horses afflicted with equine motor neuron disease (EMND): a preliminary study of a potential pathologic marker

Equine motor neuron disease (EMND) is a neurodegenerative disorder of unknown etiology affecting horses worldwide. Trans-Active Response DNA Binding Protein of 43?kDa (TDP-43) has been reported in the central nervous system (CNS) of several neurodegenerative conditions in humans including Amyotrophic Lateral Sclerosis (ALS) and assumed to play role in the disease. We examined whether horses afflicted with EMND express the TDP-43 in CNS. Ten horses with EMND and 6 controls of different ages and breed we enrolled. Detection of presence of TDP-43 protein in the CNS was analyzed by immunohistochemical staining using rabbit anti-human TARDBP (TDP-43) polyclonal antibody. Formalin fixed neuronal tissues from medulla, cervical, and lumbar spinal cord were harvested from EMND and from control horses. Sections were assigned randomly to TDP-43 treated or rabbit anti-IgG as control. Nuclear staining of TDP-43 was detected in one of the neural tissues of 75?% of EMND-positive and 0 of 0?% of control horses in the central nervous system (medulla, and/or cervical spinal cord and/or lumbar spinal cord). TDP-43 antibody was detected in the nucleus of EMND horses and no cytoplasmic staining was noted. As in ALS, there was no pattern of age clustering associated with the detection of TDP-43. This is the first report on the staining of TDP-43 in neuronal tissues of horses and suggests that TDP-43 may play a role in the pathogenesis of EMND. Further studies are needed to elucidate the etiologic role of this protein in the diseases.     

Evaluation of glucose tolerance and intestinal luminal membrane glucose transporter function in horses with equine motor neuron disease

OBJECTIVE: To confirm whether the plasma glucose concentration curve obtained during oral glucose tolerance tests (OGTTs) in horses with equine motor neuron disease (EMND) is decreased, compared with that obtained in clinically normal horses, and determine whether that decrease is a result of defective glucose metabolism or intestinal glucose transport dysfunction. ANIMALS: 8 horses with EMND and 44 matched control horses. PROCEDURE: Electromyography and OGTTs were performed in all 8 affected horses and 10 control horses. Intravenous GTTs (IVGTTs) were performed in 6 affected horses and another 11 control horses. The activity and levels of jejunal luminal membrane glucose transporter (Na+ / glucose cotransporter isoform 1 [SGLT1]) were measured in 2 affected horses and 23 control horses. RESULTS: In horses with EMND, generalized neuropathy was detected via quantitative electromyography; the mean increase in plasma glucose concentration during the OGTT was significantly decreased, compared with the value in control horses. During the IVGTT the mean increase in plasma glucose concentration was significantly lower than that of control horses. The activity and levels of SGLT1 in 2 affected horses were similar to those of control horses. Diagnosis of EMND was confirmed postmortem in all affected horses. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggest that the decreased plasma glucose curve obtained in horses with EMND during OGTTs (compared with control horses) is a result of overall enhanced glucose metabolism or abnormalities in the facilitated glucose transporters; definitive identification of the underlying mechanisms could aid in the development of appropriate treatments of EMND in horses.     

THE USE OF MAGNETIC RESONANCE IMAGING IN EVALUATING HORSES WITH SPINAL ATAXIA

To determine the accuracy of magnetic resonance imaging for diagnosing cervical stenotic myelopathy in horses, 39 horses with spinal ataxia and 20 control horses underwent clinical and neurologic examinations, cervical radiographs, euthanasia, magnetic resonance (MR) imaging of the cervical spine and necropsy. Twenty‐four horses were diagnosed with cervical stenotic myelopathy, 5 with cervical vertebral stenosis, 7 with idiopathic ataxia, 3 horses had other causes of ataxia, and 20 were controls. The MR images were assessed for spinal cord intensity changes, presence of spinal cord compression, spinal cord compression direction, shape of spinal cord, and the presence of synovial cysts, joint mice, and degenerative joint disease. The height, width, and area of the spinal cord, dural tube and vertebral canal were measured. The identification of spinal cord compression on MR images was significantly different in horses with cervical stenotic myelopathy (P < 0.02), but in the cervical stenotic myelopathy group the identification of spinal cord compression on MR images had poor to slight agreement with histopathologic evidence of compression (κ = 0.05). Horses with cervical stenotic myelopathy were more likely to have a T2 hyperintensity in the spinal cord (P < 0.05). Horses with cervical stenotic myelopathy or cervical vertebral stenosis were more likely to have degenerative joint disease than control horses or horses with other or idiopathic ataxia.     

Cardiovascular changes associated with intravenous administration of fumonisin B1 in horses

OBJECTIVE: To determine whether cardiovascular dysfunction is evident in horses with leukoencephalomalacia experimentally induced by administration of fumonisin B1. ANIMALS: 11 healthy horses of various breeds (body weight, 252 to 367 kg). PROCEDURE: Horses were randomly assigned to 3 groups and administered fumonisin B1 daily. Horses received IV injections of 0 (control horses; n = 4), 0.01 (3), or 0.20 mg (4) of fumonisin B1/kg for 7 to 28 days. Horses were examined daily for evidence of neurologic disease. When neurologic signs consistent with leukoencephalomalacia were evident, horses were anesthetized, and catheters were inserted for evaluation of the cardiovascular system. After recovery from anesthesia, hemodynamic measurements were obtained. RESULTS: Fumonisin-treated horses with clinical signs of neurologic disease had evidence of cardiovascular dysfunction manifested as decreases in heart rate, cardiac output, right ventricular contractility (assessed by measuring the maximal rate of change of right ventricular pressure), coccygeal artery pulse pressure, and pH and base excess in venous blood as well as increases in systemic vascular resistance, compared with values for control horses. Fumonisin-treated horses with and without clinical signs of neurologic disease also had higher serum and right ventricular sphinganine and sphingosine concentrations than control horses. CONCLUSIONS AND CLINICAL RELEVANCE: An association was detected among fumonisin-induced neurologic disease, increased serum and myocardial sphinganine and sphingosine concentrations, and decreased cardiovascular function in horses. Fumonisin-induced decreases in cardiovascular function may contribute to the pathophysiologic development of leukoencephalomalacia in horses.     

Plasma and liver copper values in horses with equine degenerative myeloencephalopathy.

Equine degenerative myeloencephalopathy (EDM) is a common spinal cord disease in the horse. The etiology of EDM currently is unknown. In other species, there are similarities in the clinical signs and neuropathological changes observed in EDM and in copper deficiency. The objective of this study was to determine if horses affected with EDM had low levels of plasma or liver copper. Plasma copper values were determined in 25 EDM affected horses and 35 normal horses. Liver copper levels were determined on 13 EDM affected horses and 22 normal horses. Plasma and liver copper values were not significantly lower in EDM affected horses than in control horses.     

Association between plasma vitamin E concentration and the risk of equine motor neuron disease

Equine motor neuron disease (EMND) is a neurodegenerative disorder of the somatic lower motorneurons that results in a syndrome of diffuse neuromuscular disease in the adult horse. The aetiology of this disorder is unknown, although prior studies have suggested that a deficiency in the lipid antioxidant vitamin E (α-tocopherol) contributes to the development of EMND. This paper describes a case-control study designed to investigate the association between plasma vitamin E levels and the risk of EMND for horses. Signalment, plasma vitamin E levels at the time of referral, and information relative to dietary and management practices were collected from 53 horses diagnosed with EMND and 69 controls. The mean plasma vitamin E concentration in EMND cases was significantly lower than that of control horses. After controlling for other risk factors of EMND, there was a statistically significant association between plasma vitamin E levels and EMND, with the likelihood of the disease increasing as the vitamin E concentration decreased. These findings support the reported role of vitamin E deficiency as one of the risk factors for EMND.     

Horses on pasture may be affected by equine motor neuron disease

REASONS FOR PERFORMING STUDY: Equine motor neuron disease (EMND) was diagnosed in 3 horses maintained on lush, grass-based pasture. This contrasted with North American studies which identified limited or no access to green herbage as an important risk factor for EMND. HYPOTHESIS: Grazing horses that have an apparently adequate intake of pasture herbage to meet normal equine vitamin E requirements can develop EMND. METHODS: Owners of 32 European horses diagnosed with EMND completed a questionnaire regarding intrinsic, managemental, nutritional and environmental factors that could potentially be risk factors for EMND, and also regarding clinical signs, treatments and case outcome. Plasma/serum vitamin E data for these horses were supplied by the veterinarians. No control population was studied. RESULTS: Thirteen of 32 horses (termed the 'grazing' group) had part- or full-time access to grass-based pasture at the onset of EMND (median duration at pasture 12 h/day, range 3-24 h). Five of these horses were at pasture for at least 235 h/day at the onset of EMND, 2 of which were at pasture for at least 23.5 h/day throughout the year. Despite grazing, all these horses had a low vitamin E status. The remaining 19 horses resembled those cases reported from North America, in that they had no or limited access to pasture. CONCLUSIONS AND POTENTIAL RELEVANCE: A diagnosis of EMND should not be discounted on the basis that a horse has access, even full-time, to lush grass-based pasture. Inadequate vitamin E intake was probably not the sole cause of either the EMND or the low vitamin E status in the grazing horses; the latter was probably the result of abnormal bioavailability or excessive utilisation of vitamin E.     

Assessment of copper and zinc status of farm horses and training thoroughbreds in south-east Queensland

The copper and zinc concentrations in the blood of stabled thoroughbred horses and in Australian Stock Horses mares at pasture, either late pregnant or lactating were determined by an atomic absorption spectroscopic method. The plasma concentration of the trace elements in these apparently normal horses were generally below the "normal" range. The plasma copper, caeruloplasmin copper, whole blood copper and plasma zinc concentrations in the stabled thoroughbreds were 0.76 +/- 0.19 micrograms/ml (n = 82), 0.56 +/- 0.14 micrograms/ml (n = 83), 0.75 +/- 0.18 micrograms/ml (n = 82) and 0.47 +/- 0.09 micrograms/ml (n = 83) respectively. The plasma copper and zinc concentrations of all the brood mares at pasture (pregnant and lactating) were 0.56 +/- 0.20 micrograms/ml and 0.47 +/- 0.11 micrograms/ml (n = 30). The plasma copper concentration of the pregnant group of mares (0.64 +/- 0.18 micrograms/ml; (n = 14) was greater than that of the lactating mares (0.49 +/- 0.21; (n = 16). Variation in the plasma copper concentration was also identified between stabled and farm horses, between horses of different stables and between horses of different ages. The proportion of plasma copper bound to caeruloplasmin was 73 +/- 11.8%. These low concentrations of copper and zinc in the plasma of apparently normal horses are of clinical significance since recent evidence has indicated that copper deficiency appears to promote the development of skeletal abnormalities in foals. An alternative to the use of a single plasma sample to identify the copper or zinc deficient horse was discussed.     

Elemental composition, water, and total lipid content in peripheral nerves, spinal cord and brain of healthy adult dogs

Peripheral nerves, spinal cord, and brain of healthy adult dogs were analysed biochemically for sodium, potassium, calcium, magnesium, copper, zinc, iron, aluminium, silicon, total chloride, total tissue water and total lipid content. Flame atomic absorption spectrophotometry was used for elemental quantitation of the hydrochloric acid tissue extracts. Cerebrum and spinal cord had similar values for all parameters measured. Total sodium values were higher in nerves compared to central nervous system (CNS) (brain and spinal cord), while respective values for potassium were lower. Tissue levels of calcium, iron and silicon were comparable in brain, spinal cord and nerves. However, magnesium concentrations were two to three times higher in CNS than in nerves; and the reverse was true for aluminium levels. Tissue concentrations of zinc and copper were marginally higher in CNS than in nerves.     

Correlation between severity of clinical signs and motor evoked potentials after transcranial magnetic stimulation in large-breed dogs with cervical spinal cord disease

OBJECTIVE: To evaluate use of transcranial magnetic motor evoked potentials for assessment of the functional integrity of the cervical spinal cord in large-breed dogs with cervical spinal cord disease. DESIGN: Randomized, controlled, masked study. ANIMALS: 10 healthy large-breed control dogs and 25 large-breed dogs with cervical spinal cord diseases. PROCEDURE: Affected dogs were allocated to 3 groups on the basis of neurologic status: signs of neck pain alone, ambulatory with ataxia in all limbs, or nonambulatory. Transcranial magnetic stimulation was performed on each dog with the same standard technique. Motor evoked potentials (MEP) were recorded from electrodes inserted in the tibialis cranialis muscle. Following the procedure, each dog was anesthetized and cervical radiography, CSF analysis, and cervical myelography were performed. The MEP latencies and amplitudes were correlated with neurologic status of the dogs after correction for neuronal path length. RESULTS: Mean MEP latencies and amplitudes were significantly different between control dogs and dogs in each of the 3 neurologic categories, but were not significantly different among dogs in the 3 neurologic categories. A linear association was evident between MEP latencies and amplitudes and severity of neurologic deficits; the more severe the neurologic deficits, the more prolonged the latencies and the more decreased the amplitudes. CONCLUSIONS AND CLINICAL RELEVANCE: Transcranial magnetic MEP are useful to assess severity of cervical spinal cord disease in large-breed dogs. Impairment of the functional integrity of the cervical spinal cord was found even in dogs with neck pain alone.     

Perception of Equine Practitioners Regarding the Occurrence of Selected Equine Neurologic Diseases in the Northeast Over a 10-Year Period

A survey was developed to examine the perception of equine practitioners regarding the occurrence of five equine neurologic diseases in the northeastern United States over the 10-year period between June 1, 1997 and June 1, 2007. This information was then compared with trends at Cornell University's Equine Hospital during the same time span, which in general agreed with practitioners' opinions. Equine herpes virus-1 (EHV-1) neurologic disease, equine motor neuron disease (EMND), and equine protozoal myelitis (EPM) have historic and current relevance. Results showed that the frequency of EMND and EPM has remained relatively stationary or decreased somewhat, whereas the frequency of the neurologic strain of EHV-1 may have increased slightly over the last decade. Less historical information on clinical disease associated with Borrelia burgdorferi infection (Lyme disease) and Parelaphostrongylus tenuis exists; however, results suggest that P. tenuis in the equine is presently emergent. Opinions regarding the existence and rate of occurrence of clinical borreliosis in horses appear divided. A better understanding of the frequency with which these diseases occur, as well as possible associated positive risk factors, will aid the equine practitioner in making an appropriate diagnosis in cases of neurologic disease in their equine patients.     

Concentrations of toxic metals and essential minerals in the mane hair of healthy racing horses and their relation to age

Concentrations of trace elements (As, Al, Pb, Cd, Hg, Se, Si, P, Na, K, Ca, Mg, Fe, Cu, Zn, Mn, Cr, Ni and Mn) in the mane hair obtained from 9 female and 15 male healthy racing Thoroughbred horses aged 2-5 years were analyzed by the inductively coupled plasma atomic emission spectrometry (ICP-AES) method. No significant differences between the female and male horses were observed in the mean concentrations of those minerals. Significantly positive correlations with age were observed in Cd (r=0.546, p<0.01) and Mo (r=0.733, p<0.001). Significantly negative correlations with age were observed in Hg (r= -0.726, p<0.001), Mn (r= -0.450, p<0.05) and Fe (r=-0.642, p<0.01). This reference range of trace elements in the mane hair of racing horses should be used to assess disease and the nutritional status in equine practice.     

Measurement of superoxide dismutase, diamine oxidase and caeruloplasmin oxidase in the blood of thoroughbreds

Methods were developed for the measurement of superoxide dismutase (SOD), diamine oxidase (DAO) and caeruloplasmin oxidase in the blood of thoroughbred horses. These enzymes were measured in 178 normal thoroughbreds stabled throughout the United Kingdom. The relationships between the activities of SOD, DAO and caeruloplasmin oxidase and the blood concentrations of their associated trace metals (copper, zinc and manganese) were studied in 52 of the thoroughbreds. Trace metals were measured by electrothermal atomic absorption spectrophotometry. No relationships were found between the activities of erythrocyte SOD and serum/whole blood copper, zinc and manganese, or serum DAO and serum copper or zinc concentrations. Caeruloplasmin oxidase in equine blood was found to be correlated to serum copper concentration, r = 0.695 (P less than 0.001) over the normal range. Samples from thoroughbreds with trace metal deficiency or toxicity were not available for study. The observed normal ranges for the activity of these enzymes are as follows: SOD: 50 to 200 units per ml whole blood between 5 and 95 percentiles; DAO: 0.1 to 28.5 units per litre (means = 14.8, SD 7.1) and caeruloplasmin oxidase; 11.6 to 35.8 units per ml (means = 23.7, SD 6.0). For numerical simplicity, the activity of DAO is given in units per litre, compared to units per ml for caeruloplasmin oxidase and SOD.     

Medical management of spinal cord disease

In spinal cord disease of horses, a complete history, neurologic examination, and adjunctive diagnostic procedures are very helpful in establishing a tentative diagnosis; however, a definitive diagnosis may be difficult or impossible to establish antemortem. Medical management should be initiated with full consideration of possible etiologies and knowledge of the effects and consequences of medical therapies. This article discusses the drugs commonly used in the management of spinal cord disease and the rationale for their use.     

Reduced tongue tone associated with degeneration of the hypoglossal nerve nucleus in a horse with equine motor neuron disease

Equine motor neuron disease (EMND) is a condition characterised by generalised weakness and muscle atrophy associated with degeneration of motor neurons in the ventral horns of the spinal cord. Despite the frequent detection of cranial nerve nuclei pathology during post mortem examination, associated clinical signs are rarely reported. This report describes a case of EMND in a pony gelding that presented with clinical signs of diffuse neuromuscular weakness associated with marked flaccidity of the tongue, making differentiation from similar neuromuscular conditions, particularly botulism, extremely challenging.     

A multicenter case-control study of risk factors for equine protozoal myeloencephalitis

OBJECTIVE: To identify risk factors for equine protozoal myeloencephalitis (EPM) among horses examined at 11 equine referral hospitals. DESIGN: Case-control study. ANIMALS: 183 horses with EPM, 297 horses with neurologic disease other than EPM (neurologic controls), and 168 horses with non-neurologic diseases (non-neurologic controls) examined at 11 equine referral hospitals in the United States. PROCEDURES: A study data form was completed for all horses. Data were compared between the case group and each of the control groups by means of bivariate and multivariate polytomous logistic regression. RESULTS: Relative to neurologic control horses, case horses were more likely to be > or = 2 years old and to have a history of cats residing on the premises. Relative to non-neurologic control horses, case horses were more likely to be used for racing or Western performance. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that cats may play a role in the natural epidemiology of EPM, that the disease is less common among horses < 2 years of age relative to other neurologic diseases, and that horses used for particular types of competition may have an increased risk of developing EPM.     

Neuroaxonal dystrophy of the accessory cuneate nucleus in horses

Data were collected from 37 horses with a neurologic disability and compared to a group of 34 normal horses. Affected horses had neuroaxonal dystrophy, gliosis, vacuoles, and sometimes pigment localized to the accessory cuneate nuclei with minimal or no changes in the spinal cord and no changes in the proximal peripheral nerves. The focal nature of the change and usual absence of significant light microscopic spinal cord or peripheral nerve changes are different than previously described equine neuropathologic conditions.     

Immunoconversion against Sarcocystis neurona in normal and dexamethasone-treated horses challenged with S. neurona sporocysts

Equine protozoal myeloencephalitis is a common neurologic disease of horses in the Americas usually caused by Sarcocystis neurona. To date, the disease has not been induced in horses using characterized sporocysts from Didelphis virginiana, the definitive host. S. neurona sporocysts from 15 naturally infected opossums were fed to horses seronegative for antibodies against S. neurona. Eight horses were given 5x10(5) sporocysts daily for 7 days. Horses were examined for abnormal clinical signs, and blood and cerebrospinal fluid were harvested at intervals for 90 days after the first day of challenge and analyzed both qualitatively (western blot) and quantitatively (anti-17kDa) for anti-S. neurona IgG. Four of the challenged horses were given dexamethasone (0.1mg/kg orally once daily) for the duration of the experiment. All challenged horses immunoconverted against S. neurona in blood within 32 days of challenge and in CSF within 61 days. There was a trend (P = 0.057) for horses given dexamethasone to immunoconvert earlier than horses that were not immunosuppressed. Anti-17kDa was detected in the CSF of all challenged horses by day 61. This response was statistically greater at day 32 in horses given dexamethasone. Control horses remained seronegative throughout the period in which all challenged horses converted. One control horse immunoconverted in blood at day 75 and in CSF at day 89. Signs of neurologic disease were mild to equivocal in challenged horses. Horses given dexamethasone had more severe signs of limb weakness than did horses not given dexamethasone; however, we could not determine whether these signs were due to spinal cord disease or to effects of systemic illness. At necropsy, mild-moderate multifocal gliosis and neurophagia were found histologically in the spinal cords of 7/8 challenged horses. No organisms were seen either in routinely processed sections or by immunohistochemistry. Although neurologic disease comparable to naturally occurring equine protozoal myeloencephalitis (EPM) was not produced, we had clear evidence of an immune response to challenge both systemically and in the CNS. Broad immunosuppression with dexamethasone did not increase the severity of histologic changes in the CNS of challenged horses. Future work must focus on defining the factors that govern progression of inapparent S. neurona infection to EPM.