Concentrations of trace minerals in the spinal cord of horses with equine motor neuron disease

OBJECTIVE: To compare concentrations of trace minerals in the spinal cord of horses with equine motor neuron disease (EMND) with those of horses without neurologic disease (control horses). ANIMALS: 24 horses with EMND and 22 control horses. PROCEDURE: Spinal cord trace mineral concentrations in horses with EMND and control horses were analyzed by use of inductively coupled plasma atomic emission spectroscopy (calcium, phosphorus, sodium, potassium, magnesium, copper, iron, manganese, nickel, zinc, aluminum, cobalt, and chromium), atomic absorption spectrophotometry (lead and cadmium), flameless atomic absorption (mercury), and fluorometry (selenium). RESULTS: Copper concentration was significantly higher in the spinal cord of horses with EMND, compared with control horses; spinal cord concentrations of all other trace minerals were similar between groups. CONCLUSION AND CLINICAL RELEVANCE: Among spinal cord trace minerals investigated in the study, only copper concentrations were significantly different between horses with EMND and horses without neurologic disease, which suggests that copper may be involved in the pathogenesis of EMND. An hypothesis of oxidative injury in this disease is supported by the finding of increased copper concentrations in the spinal cord and by low vitamin E concentrations reported by other researchers.     

Gliomatosis cerebri in two dogs

A 3.5 yr old Saint Bernard was evaluated for nonambulatory tetraparesis and cranial nerve dysfunction, and a 7 yr old rottweiler was evaluated for progressive paraparesis. Clinical signs of left-sided vestibular and general proprioceptive ataxia and cranial nerve VII dysfunction in the Saint Bernard suggested a lesion affecting the brain stem. Signs in the rottweiler consisted of general proprioceptive/upper motor neuron paraparesis, suggesting a lesion involving the third thoracic (T3) to third lumbar (L3) spinal cord segments. MRI was normal in the Saint Bernard, but an intra-axial lesion involving the T13-L2 spinal cord segments was observed in the rottweiler. In both dogs, the central nervous system (CNS) contained neoplastic cells with features consistent with gliomatosis cerebri (GC). In the Saint Bernard, neoplastic cells were present in the medulla oblongata and cranial cervical spinal cord. In the rottweiler, neoplastic cells were only present in the spinal cord. Immunohistochemistry disclosed two distinct patterns of CD18, nestin, and vimentin staining. GC is a rarely reported tumor of the CNS. Although GC typically involves the cerebrum, clinical signs in these two dogs reflected caudal brainstem and spinal cord involvement.     

A case of equine motor neuron disease (EMND)

We report a case of EMND in a heavy horse that was bred and trained in Hokkaido, Japan. Clinical symptoms included severe ataxia of all four limbs, tilted head, lethargy, and flaccid lips. Numerous axonal degenerations and swellings were observed in nuclei, mostly in the cerebellar dentate nucleus and the nucleus of the hypoglossal nerve, and in the ventral horn of the spinal cord. In the ventral horn of the spinal cord, neuronal degeneration, swelling, and/or necrosis were observed sporadically. The case was diagnosed as EMND from the clinical symptoms and pathological findings.     

Endoscopic anatomy of the cervical vertebral canal in the horse: a cadaver study

Reason for performing study: Localisation of spinal cord compression in horses with cervical vertebral stenotic myelopathy is inexact. Vertebral canal endoscopy has been used in man to localise spinal cord lesions and has the potential to become a useful diagnostic technique in horses. Objective: To establish a surgical approach via the atlanto‐occipital space to the cervical vertebral canal in equine cadavers and describe the endoscopic anatomy of the cervical epidural and subarachnoid spaces. Methods: The cadavers of 25 mature horses were used to assess 3 surgical methods to approach the cervical vertebral canal, including 2 minimally invasive and one open technique. Once the approach had been made, a flexible videoendoscope was inserted into the epidural space (epiduroscopy) or the subarachnoid space (myeloscopy) and advanced caudally until the intervertebral space between C7 and T1 was reached. Results: The epidural and subarachnoid spaces could not be accessed reliably using the minimally invasive techniques. Furthermore, damage to the nervous tissues was a frequent complication with these procedures. The open approach allowed successful insertion of the videoendoscope into the epidural and subarachnoid spaces in all horses and no inadvertent damage was observed. Anatomical structures that could be seen in the epidural space included the dura mater, nerve roots, fat and the ventral internal vertebral venous plexus. In the subarachnoid space, the spinal cord, nerve roots, blood vessels, denticulate ligaments and external branch of the accessory nerve were seen. Conclusions: Using the open approach, epiduroscopy and myeloscopy over the entire length of the cervical vertebral canal are possible in the mature horse. Potential relevance: Cervical vertebral canal endoscopy may become a valuable tool to localise the site of spinal cord injury in horses with cervical vertebral stenotic myelopathy and could aid in the diagnosis of other diseases of the cervical spinal cord.     

Lesions in the central nervous system associated with perinatal lamb mortality

OBJECTIVE: To identify and describe the occurrence of neurological lesions that could have an effect on lamb mortality. PROCEDURE: The central nervous system was investigated macroscopically (n = 92) and microscopically (n = 72) in lambs dying in the perinatal period during 3 years in flocks of adult Corriedale ewes. The central nervous system was removed intact and samples of cerebral cortex, basal ganglia, thalamus, hippocampus, mesencephalon, cerebellar cortex, medulla oblongata, and cervical spinal cord were scored microscopically for the severity of neuronal dead, cytotoxic and perivascular oedema, and haemorrhage. RESULTS: Neurologic findings between birth and 6 days included haemorrhages in meninges, brain congestion and oedema, neuronal ischemic necrosis, intraparenchymal haemorrhages in medulla oblongata and cervical spinal cord, parasagittal cerebral necrosis, and periventricular leukomalacia. No significant lesions were found in anteparturient deaths or in those aged between 7 and 16 days. Oedema was more severe in the brain than in other regions of the central nervous system. Ischaemic neurons first appeared 24 hours post partum, increased linearly in number between 48 hours and 5 days post partum, and had a laminar distribution in the cerebral cortex, indicating a hypoxic-ischemic encephalopathy. Haemorrhages were most severe in the gray matter of medulla oblongata and cervical spinal cord, suggesting trauma due to instability of atlantoaxialis joint. CONCLUSION: Lesions in the central nervous system can explain most deaths at birth and within 6 days of birth. The lesions were hypoxic-ischemic and appeared to be related to birth injury.     

Diagnostic exercise: astrocytoma with involvement of medulla oblongata, spinal cord, and spinal nerves in a raccoon (Procyon lotor)

Neoplasms affecting the central and peripheral nervous systems of wild animals are extremely rare. Described are clinical signs and pathologic and immunohistochemical findings in an adult female raccoon (Procyon lotor) with an astrocytoma that involved medulla, cervical spinal cord, and roots of the cervical spinal nerves. Microscopically, the neoplastic cells revealed some pleomorphism but generally had fusiform morphology and showed moderate numbers of mitotic figures. Glial fibrillary acidic protein was demonstrated within the neoplastic cells by immunohistochemistry. This appears to be the only reported case of astrocytoma that involved multiple anatomic sites in the central nervous system of this raccoon.     

Ataxia and weakness associated with fourth ventricle vascular anomalies in two horses

Two adult horses with progressive neurologic signs were examined clinically and at necropsy. Both horses had signs of progressive ataxia and weakness, clinically diagnosed as spinal cord in origin. Differential diagnoses for cervical spinal ataxia in horses included cervical vertebral malformation, equine degenerative myeloencephalopathy, equine herpes-virus-I myeloencephalopathy, and equine protozoal myeloencephalopathy. Necropsy findings in both horses were similar and consisted of a large hematoma in the fourth ventricle, with upward compression of the cerebellum and downward compression of the pons and rostral portion of the medulla.     

Chronic cervical compressive myelopathy in horses: patterns of astrocytosis in the spinal cord

The distribution and morphology of fibrous astrocytes in the cervical spinal cord of normal horses and horses with chronic compressive myelopathy were demonstrated using immunohistochemical staining for glial fibrillary acidic protein. In the spinal cord from normal horses, astrocytes with stellate cell bodies and short processes were irregularly distributed in grey matter. In the white matter, their cell bodies were small and angular in areas adjacent to grey matter and larger and more stellate-shaped in the subpial area. Astrocyte processes were fine, and evenly distributed in a predominantly radial pattern in transverse sections of cord. Gliosis was marked in the spinal cords of horses with cervical compressive myelopathy. In the grey matter at the level of compression astrocytes were often enlarged and rounded, with short, blunt processes, but the gliosis was generally mild. In the white matter, gliosis was obvious in areas of nerve fibre swelling and degeneration at the level of compression and in areas of ascending and descending Wallerian degeneration. The fine radial pattern of astrocyte fibres was replaced by a dense, irregular arrangement. Gliosis persisted in the cords of chronically affected horses after active nerve fibre degeneration had subsided. The areas of gliosis coincided with the areas of Marchi staining for degenerating myelin and with areas of myelin loss in osmium tetroxide post-fixed tissue. Histological observations were consistent with astrocytes replacing areas of extracellular space that remained after nerve fibre degeneration. it is concluded that astrocytic gliosis is a prominent and persistent alteration of the spinal cord of horses with chronic cervical compressive myelopathy.     

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.     

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.     

Localisation and expression of a myelin associated neurite inhibitor,Nogo-A and its receptor Nogo-receptor by mammalian CNS cells

Axon regeneration failure in the adult mammalian central nervous system (CNS) is partly due to inhibitory molecules associated with myelin. The Nogo receptor (NgR) plays a role in this process through an extraordinary degree of cross reactivity with three structurally unrelated myelin-associated inhibitory ligands namely; Nogo-A, myelin associated glycoprotein (MAG) and oligodendrocyte myelin glycoprotein (OMgp). The major aim of the study was to investigate and explore the cellular localisation and expression pattern of NgR and Nogo-A in the mammalian nervous system. We therefore generated a rabbit polyclonal anti-NgR antibody from the leucine rich repeat (LRR) No. 9 domain of the NgR polypeptide chain. Together with a commercially available polyclonal antibody specific for NgR, and in conjunction with double labeling immunofluorescence methods on cryosections and cell cultures, NgR immunoreactivity was observed in the CNS and dorsal root ganglia (DRG). In cellular populations, it was confined to neuronal cell bodies and their processes. NgR was also localised on the surface of extending DRG intact axons and growth cones in live staining experiments. Nogo-A, a member of the reticulon family protein, was widely distributed in the mammalian brain, spinal cord, and DRG. Intense Nogo-A immunoreactivity was also detected in oligodendrocyte cell bodies and their myelin sheaths in nerve fibre tracts of the CNS. Furthermore, numerous populations of neurons in the brain and spinal cord expressed Nogo-A to a variable extent in their cell bodies and neurites, suggesting additional, as-yet-unknown, functions of this protein. These results confirm results obtained by other researchers with different sets of antibodies. However, they also raise the question of the mechanism and circumstances under which NgR interacts with Nogo-A, as the latter appears to be confined to the cytoplasm and can therefore not be expected to bind NgR on the axon surface.     

Serological study of an outbreak of paresis due to equid herpesvirus 1 (EHV-1).

Six cases of paresis occurred in a Swedish stud with 48 mares and a stallion. Complement-fixation tests revealed a recent infection with EHV-1 in most horses of the stud. Serumneutralisation tests showed rapid antibody-titre increases during the course of the disease. This type of antibody response was interpreted as induced by reinfection or, possibly, recurrent infection. Two diseased mares were sacrificed. No virus could be isolated from their central nervous system (CNS), liver or spleen, but there is a presumptive evidence for the presence of an antigen specific to EHV-1 in the CNS and liver. Neutralising antibodies to EHV-1 were demonstrated in the liver and kidneys following elution by acidification of the tissues. No such antibodies could be demonstrated in the brain and spinal cord. A possible reason for this failure is discussed.     

Localisation and expression of a myelin associated neurite inhibitor, Nogo-A and its receptor Nogo-receptor by mammalian CNS cells

Axon regeneration failure in the adult mammalian central nervous system (CNS) is partly due to inhibitory molecules associated with myelin. The Nogo receptor (NgR) plays a role in this process through an extraordinary degree of cross reactivity with three structurally unrelated myelin-associated inhibitory ligands namely; Nogo-A, myelin associated glycoprotein (MAG) and oligodendrocyte myelin glycoprotein (OMgp). The major aim of the study was to investigate and explore the cellular localisation and expression pattern of NgR and Nogo-A in the mammalian nervous system. We therefore generated a rabbit polyclonal anti-NgR antibody from the leucine rich repeat (LRR) No. 9 domain of the NgR polypeptide chain. Together with a commercially available polyclonal antibody specific for NgR, and in conjunction with double labeling immunofluorescence methods on cryosections and cell cultures, NgR immunoreactivity was observed in the CNS and dorsal root ganglia (DRG). In cellular populations, it was confined to neuronal cell bodies and their processes. NgR was also localised on the surface of extending DRG intact axons and growth cones in live staining experiments. Nogo-A, a member of the reticulon family protein, was widely distributed in the mammalian brain, spinal cord, and DRG. Intense Nogo-A immunoreactivity was also detected in oligodendrocyte cell bodies and their myelin sheaths in nerve fibre tracts of the CNS. Furthermore, numerous populations of neurons in the brain and spinal cord expressed Nogo-A to a variable extent in their cell bodies and neurites, suggesting additional, as-yet-unknown, functions of this protein. These results confirm results obtained by other researchers with different sets of antibodies. However, they also raise the question of the mechanism and circumstances under which NgR interacts with Nogo-A, as the latter appears to be confined to the cytoplasm and can therefore not be expected to bind NgR on the axon surface.     

Clinical, pathological and toxicological findings of a iatrogenic selenium toxicosis case in feeder pigs

Histopathological and toxicological analyses confirmed a clinical diagnosis of selenium (Se) intoxication in pigs from a farm in Spain. After an initial episode of diarrhoea, animals presented both dermatological and neurological signs; the most obvious sign was a marked hind limb paresis. Cutaneous lesions consisted on diffuse alopecia, multifocal skin necrosis and coronary band necrosis of the hooves. Central nervous system lesions involved the cervical and lumbar intumescences of the spinal cord and consisted of a severe, bilateral symmetrical poliomyelomalacia of the ventral horns; pons and medulla oblongata also presented lesions of polioencephalomalacia. Analyses of feed and sera from clinically affected pigs revealed a marked increase in Se concentration. Clinical investigations indicated that a failure in Se dosage in feed was the cause of the toxicosis.     

兔中枢神经内Orexin B免疫阳性神经纤维的分布

采用免疫组织化学方法研究了青紫蓝兔中枢神经系统（CNS）内orexin B免疫阳性纤维的分布。结果显示，orexin B免疫阳性神经纤维广泛分布于CNS内，在端脑分布于大脑皮质、隔核和杏仁核；在间脑分布于丘脑、下丘脑、上丘脑和垂体，主要分布于丘脑中线核和下丘脑；在中脑分布于中脑中央灰质、前丘、后丘、脚间核和中缝核；在脑桥分布于蓝斑、被盖背侧核和中缝核；在延髓分布于最后区、孤束核和迷走神经背侧运动核；在小脑和脊髓也有分布。兔CNS内orexin B免疫阳性纤维的分布与orexin A的基本相似，本试验结果为理解orexin B在兔体内的作用部位提供了形态学资料。     

嗅鞘细胞生物学特性及其在脊髓再生中应用的研究

近年来研究证明嗅球及嗅神经组织可以分离出嗅神经鞘细胞(olfactory ensheathing cells,OECs)，OECs具有中枢神经系统（CNS）星形胶质细胞和外周雪旺式细胞的特性，移植后的OCEs能够在宿主损伤的脊髓组织进行迁移，能够引导支持损伤的神经元突起再生、生长、延伸。而且还可以通过转基因技术使其携带某些促进神经元再生的外源性基因，表达一些促神经再生的分子，从而改善神经损伤的内环境，诱导损伤的脊髓神经元再生，使得受损脊髓再生的潜力得以发挥，达到脊髓再生和功能恢复的目的。本文就OECs的生物学特性及其在脊髓损伤和CNS的基因治疗进行综述。     

Equine motor neuron disease; a preliminary report

A spontaneous motor neuron disease or neuronopathy was identified in 10 horses from the northeastern United States. Signs of generalized weakness, muscle fasciculations, muscle atrophy and weight loss progressed over 1 to several months in young and old horses of various breeds. Pathologic studies revealed that degeneration and loss of motor neurons in the spinal cord and brain stem resulted in axonal degeneration in the ventral roots and peripheral and cranial nerves and denervation atrophy of skeletal muscle. Many spinal neurons were swollen, chromatolytic and contained neurofilamentous accumulations. Other cell bodies were shrunken and undergoing neuronophagia and some were lost and replaced by glia. This fatal equine motor neuron disease has not been reported previously and its cause has not been determined. The progressive weakness and wasting and the neuronal degenerative changes in these horses were similar to those described in people with sporadic amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig's disease.     

Use of magnetic motor-evoked potentials in horses with bilateral hind limb ataxia

OBJECTIVE: To determine the usefulness of magnetic motor-evoked potentials (MMEPs) for assessing the integrity of the cervical, thoracic, and thoracolumbar spinal cord in horses with bilateral hind limb ataxia. ANIMALS: 9 horses and 1 donkey with bilateral hind limb ataxia of various degrees. PROCEDURE: The motor cortex was stimulated magnetically, and MMEPs were recorded bilaterally from the extensor carpi radialis and cranial tibial muscles. RESULTS: In 5 horses and 1 donkey, MMEPs with normal onset latencies and peak-to-peak amplitude were recorded from the extensor carpi radialis muscles, whereas abnormal onset latencies and peak-to-peak amplitudes were recorded from the cranial tibial muscles. In these animals, a spinal cord lesion in the thoracic or thoracolumbar segments was suspected. In 4 horses, onset latencies and peak-to-peak amplitude of MMEPs recorded from the extensor carpi radialis and cranial tibial muscles were abnormal. In these horses, a cervical spinal cord lesion was suspected. CONCLUSIONS AND CLINICAL RELEVANCE: Transcranial magnetic stimulation can be considered a valuable diagnostic tool for assessing the integrity of the spinal cord, and MMEPs may be used for differentiating thoracic or thoracolumbar spinal cord lesions from mild cervical spinal cord lesions that cause ataxia in the hind limbs only.     

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.     

Equine herpesvirus type 1 (EHV-1) myeloencephalopathy: a case report

An outbreak of neurological disease occurred in a well-managed riding school. Ataxia and paresis were observed in several horses, five of which became recumbent and were euthanized. Post-mortem analysis revealed scattered haemorrhages along the spinal cord, that were reflected by multiple haemorrhagic foci on formalin-fixed sections, with the thoracic and lumbar segments being the most affected. Pathohistologically, perivascular mononuclear cuffing and axonal swelling, especially in the white matter, were evident. Parallel to the course of disease, alterations in myelin sheets and activation of astrocytes and microglial cells were also observed. Virological findings confirmed an acute equine herpesvirus type 1 infection and virus was isolated from the spinal cord of a 26-year-old mare.