A morphometric and stereological study on cervical spinal cord segment of goose

In this study, volume density of white matter and grey matter areas of cervical segment of spinal cord in adult geese weighing 3–4 kg was examined using stereological methods. 10 geese were used as material without sex discrimination. All animals underwent perfusion with 10% buffered formaldehyde. Following the perfusion, animals were kept in 10% formaldehyde for 1 week. Geese were then dissected. Cervical area of spinal cord was revealed removing cervical spine. Tissue samples were obtained from each segment of cervical area. 5 μm thick cross‐sections were taken from these tissue samples via microtome. Series of cross‐sections were obtained by sampling in the ratio of 1/250 including 12 cross‐sections from each cervical segment of every animal. Cross‐sections were stained by haematoxylin eosin. They were photographed under microscope. Volume density (volume fractions) of both whole tissue and white matter and grey matter parts in each cervical segment of spinal cord were calculated using Cavalieri’s Principle. In the study, total volume of cervical segment, volume of white matter and grey matter, and ratios of these volumes one another were assessed in goose.     

A stereological study on calculation of volume values regarding lumbosacral segments of quails

In the study, total segment, grey matter and white matter volume values of lumbosacral segment of spinal cord in quails (Coturnix coturnix japonica) were stereologically examined. Regardless of sex, six adult quails were used as material. After the materials were perfused, they were dissected and their spinal cords were uncovered. Tissue samples were taken from each lumbosacral segment of spinal cord. After the tissue fixation process, 6–7 sections having a 5‐µm thickness were separated in every 50th section as from the first section, where the tissue was seen, among tissue samples of each segment. These segments were stained with haematoxylin–eosin staining. They were photographed via a microscope. By using the Cavalieri's principle, the volume values of both the total segment, grey matter (GM) and white matter (WM) in each lumbosacral segment were separately calculated. All these calculations were performed by using SHTEREOM 1.5 program. As a result, total volume, grey matter and white matter volume values of lumbosacral segments and white matter/total volume, grey matter/total volume, and grey matter/white matter volume fractions were obtained.     

A Morphological and Stereological Study on Cervical Segment of Spinal Cord of Quails

In this study, volume densities of white and grey matters of cervical segments of spinal cords of quail were investigated stereologically. In both sexes, mature, six quails were used as material of this study. All animals were fixed by perfusing in 10% buffered formalin. Tissue specimens were obtained from cervical spinal cords. These tissue specimens were cut every fiftieth section at 5 μm thickness by a microtome. And mean six or seven sections were examined from every block by this method at microscope. After that, these sections were stained by haematoxylin eosin and photographed. Densities of volumes of all tissues of cervical segments of whole spinal cords and white and grey matters were calculated with principle of Cavalieri. As a result, total volume of spinal cord, volumes of white and grey matters of cervical segment and volume rates of white and grey matters were calculated.     

Morphometry of cervical spinal cord in cat using design-based stereology

The spinal cord harbours nerve fibres that facilitate reflex actions and that transmit impulses to and from the brain. The cervical spinal cord is an area of particular interest in medicine and veterinary due to frequent pathologic alterations in this region. This study describes the morphometric features of the cervical spinal cord in cat using design-unbiased stereological methods. The cervical spinal cords of four male cats were dissected and samples were taken according to systematic uniform random sampling. Each sample was embedded in agar and cut into 60-µm thick sections and stained with cresyl violet 0.1% for stereological estimations. The total cervical spinal cord volume obtained by the Cavalieri estimator was 2,321.21 ± 285.5 mm³. The relative volume of grey matter and white matter was 23.8 ± 1.3% and 76.1 ± 1.3%. The dorsal horn and ventral horn volume were 12.3 ± 1.2% and 11.4 ± 0.7% of the whole cervical spinal cord. The volume of central canal was estimated to 3.8 ± 1 mm³. The total number of neurons was accounted 3,405,366.2 ± 267,469.4 using the optical disector/fractionator method. The number of motoneurons and interneurons was estimated to be 1,120,433.2 ± 174,796.7 and 2,284,932.9 ± 127,261.5, respectively. The average volume of the motoneurons and interneurons was estimated to 1980 µm³ and 680 µm³, respectively, using the spatial rotator method. This knowledge of cat spinal cord findings may serve as a foundation as a translational model in spinal cord experimental research and provide basic findings for diagnosis and treatment of spinal cord disorders.     

Anaplastic astrocytoma in the spinal cord of an African pygmy hedgehog (Atelerix albiventris)

A 2-year-old, female hedgehog presented with an 8-month history of progressive, ascending paresis/paralysis and was tentatively diagnosed with wobbly hedgehog syndrome. She died awaiting further diagnostic tests, and the owners consented to postmortem examination. Grossly, the bladder was large and flaccid and the cervical and lumbar spinal cord were regionally enlarged, light grey, and friable with multifocal hemorrhages. The thoracic spinal cord was grossly normal. Microscopically all regions of the spinal cord had similar changes, although the cervical and lumbar sections were most severely affected. These regions were completely effaced by a moderately cellular infiltration of highly pleomorphic polygonal to spindle shaped cells, mineralization, and necrosis, which were most consistent with anaplastic astrocytoma. The thoracic spinal cord white matter was similarly infiltrated by the neoplastic cells, with perivascular extension into the otherwise normal grey matter. A diagnosis of anaplastic astrocytoma was confirmed using immunohistochemical stains that were positive for glial fibrillary acidic protein and S100.     

A morphological and stereological study on brain,cerebral hemispheres and cerebellum of New Zealand rabbits

The aim of this research was to determine brain, cerebral hemispheres and cerebellum volume and volume ratios by using stereological methods and investigate morphological differences between female and male New Zealand rabbits. The study was applied on 14-month old (10 male and 10 female) New Zealand rabbits. The materials removed from the cavum cranii using dorsal approach. After following routine histological procedure, paraffin blocks were prepared and cut every seventieth section at 10 μm thickness. Slides were stained with Crossmon's triple stain and photographed. The sectional images obtained were transferred to ImageJ program to estimate grey and white matter volume on cerebral hemispheres and cerebellum with principle of Cavalieri. According to results, there was no asymmetry on the left and right cerebral hemispheres of New Zealand rabbits. In the total hemisphere volume calculated by Cavalieri principle, grey and white matter ratio was 81.57% and 18.43% in female, 82.80% and 17.20% in male. It was found that the white matter was significantly higher in females than males in cerebral hemispheres (p < .05). Also, it was found that grey and white matter ratio in total cerebellum volume was 67.82% and 32.18% in female, 67.94% and 32.06% in male respectively. It was determined that the females' white matter was larger than male rabbits in cerebellum (p < .05). It is thought that morphometric data obtained from this study will contribute to the existing anatomical knowledge and also considered as reference values in the clinical sciences.     

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.     

Anatomical Studies on the Spinal Cord Segments of the Impala

The anatomy of the spinal cord segments was studied and recorded for the impala. The root attachment lengths were greatest at C₃, T₁₀ and L₃ cord segment levels in the respective regions. As to the root emergence length the greatest lengths were observed at C7, T_]0> L5 and S₁ cord segment levels respectively. The interroot interval was longest at C₂, T₈ and L₁ segments respectively. The longest cord segments were C2, T13, L₂ and S₂ segments. The widest cord segments of their respective regions were C₇, T₁, L5 and S₁ cord segments. As to segment volume C3, T₁₃, L₂ and S₁ were the most voluminous cord segments in the respective cord regions. Statistical analysis revealed a high correlation among all of the study parameters suggesting a high degree of multicolinearity. Gross anatomical relationships concerning the location of the spinal cord segments with respect to the vertebrae were studied. The cord segments C], T_s–T₄ and Li–L₃ were within their vertebral limits. In the impala the spinal cord terminated at the midlevel of S₄ vertebra.     

Investigations on the Postnatal Development of the Macroscopic Proportions and the Topographic Anatomy of the Feline Spinal Cord

The aim of this work was to investigate the postnatal development of the feline spinal cord. Our study showed that the main period of growth leading to the cervical and lumbar enlargements begins after birth and is completed at the age of 5–6 months. Comparing the relationship between the length of the spinal cord and the vertebral column, we found that in contrast to the adult cat, in the newborn cat, length, area and volume of segments show similar values along the spinal cord. This also applied to the length of the vertebrae. Due to a heterogeneous growth, not all segments of the spinal cord end up situated cranial to their corresponding vertebrae. As a consequence, the end of the conus medullaris is still located within the sacral canal in animals older than 2 months. These findings strongly propose that injections into the vertebral canal of the cat have to be performed caudal to the sacral vertebrae.     

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.     

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.     

Canine spinal cord neuron and axon myelin sheath morphometry

This inedited morphometric study has been developed from healthy canine spinal cord neuron cytoplasm and nucleus, and white matter axonal myelin sheath, from cervical, thoracic and lumbar regions. For the morphometric study, the parameters were area, perimeter, maximum and minimum diameters and roundness for neurons and myelin thickness for axon. For each parameter, 300 neurons were analysed. The results revealed that lumbar neurons had the highest mean values for the analysed parameters, indicating the presence of large neurons in this region, with large axons as a result of myelin thickness, which is proportional to axon calibre. We conclude that these morphometric results can contribute for the establishment of normal patterns, for canine spinal cord cervical, thoracic and lumbar segments.     

Fibrocartilaginous embolic myelopathy in a calf.

A 5-month-old Angus heifer with a history of acute hindlimb paresis that quickly progressed to lateral recumbency was necropsied. Gross lesions included a 6-cm segment of gray to brown discoloration and softening of the right ventrolateral spinal cord between T2 and T3. Microscopically, there was liquefactive necrosis of ventrolateral white and gray matter, and multiple intravascular emboli partially or completely occluded many intralesional and adjacent spinal and meningeal arteries and veins. Emboli were alcian blue positive, consistent with fibrocartilage of the nucleus pulposus of the intervertebral disk. No gross abnormalities were detected in the vertebrae or intervertebral disks. Fibrocartilaginous embolic myelopathy appears to be very rare in cattle; however, it should be considered in cases of acute, nonprogressive spinal cord dysfunction.     

Morphometric and volumetric analysis of the cervical spinal cord and vertebral canal using computed tomography images in normal Van cats

This study was performed on Van cats to determine the volumetric, morphometric and surface area measurement values of their cervical spinal cords (SC) and vertebral canals (CC) and the ratios between their dimensions using computed tomography (CT) images. The study also aims to reveal any biometric differences in these values between the two sexes. Spinal cord and vertebral canal CT images of 16 healthy adult Van cats (8 males and 8 females) were used in the study. First, three measurement points were selected for each vertebra on their cranial, medial and caudal sections along the sagittal axis. Next, the morphometric values were obtained using the transversal images of these measurement points. The surface areas for the SC and CC were calculated using the stereological planimetry method. The Cavalieri's principle was then used to calculate the relevant anatomic structure volumes. The obtained values were then statistically analysed. SC and CC were found to be larger in males in general, while ‘SC cranial/CC cranial’ and ‘SC medial/CC medial’ dimensions and volume ratios were found to be larger in female cats on average. Bodyweight and age values were found to have a negative correlation with SC and CC ratio, but the correlation was statistically insignificant. All SC and CC surface area and volume measurements were found to be higher in male cats (p < .05). We believe the results obtained by this study will provide valuable insight into veterinary clinicians in evaluating pathological lesions in the cervical spinal cord and vertebral canal CT scans.     

Spinal projections of cat primary afferent fibers innervating caudal facet joints

The spinal projections of afferent fibers innervating the facet joints between caudal vertebrae were examined by the use of anterograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Experiments were performed on 5 adult cats in which spinal dorsal roots below the 2nd sacral segment (S2) on the right side were cut. Injections of WGA-HRP into the caudal facet joints gave rise to extensive cranio-caudal distribution of WGA-HRP positive products along the spinal cord, indicating that many afferent fibers innervating unilateral facet joints terminate bilaterally in laminae I-II, V-VI and X of the thoracic, lumbar, sacral and caudal spinal cord. These afferent fibers may convey a series of sensory information from the caudal facet joints to the spinal cord.     

Verminous (Strongylus vulgaris) myelitis in a donkey

A fifth stage Strongylus vulgaris migrated through the spinal cord of a 2-year-old, male donkey resulting in progressive paraparesis and then tetraplegia. A profound neutrophilic pleocytosis was detected on analysis of cerebrospinal fluid. The parasite appeared to have entered the mid-lumbar spinal cord, migrated to the cranial thoracic segments, exited, then re-entered the spinal cord a few segments craniad. It then traveled further cranially and was found in the third cervical spinal cord segment. Some parts of the lesion were remarkably free from tissue necrosis, hemorrhage and inflammation. Severe granulomatous myelitis with hemorrhage and necrosis were seen at other sites. The latter were quite similar to lesions seen in equine protozoal myeloencephalitis.     

The Effects of Aortic Occlusion on Transcranially Induced Evoked Potentials in the Dog

Evoked potentials were produced by anodal stimulation over the motor cortex in six dogs. Potentials were recovered from the cranial thoracic and caudal lumbar portions of the spinal cord, and the radial and sciatic nerves. Evoked potential averages were recorded every 1.5 minutes during 40 minutes of aortic occlusion and during 40 minutes of reperfusion. Mean amplitudes of evoked potentials recovered from the caudal lumbar spinal cord decreased to 50% of original values at minute 12.2. Upon release of occlusion, the evoked potentials returned to baseline levels and remained there throughout the period of reperfusion. Sciatic nerve amplitudes decreased to 50% of original values at minute 4.5. In no subject could wave forms be recovered after minute 9.0. Upon release of occlusion, the evoked potentials returned to baseline levels and above, then deteriorated to 29 +/- 12% after 40 minutes of reperfusion. We concluded that transcranially induced evoked potentials were highly sensitive to spinal cord ischemia. Evoked potentials recovered from the sciatic nerve were consistent with functional grey matter immediately upon reperfusion, but deteriorated during reperfusion.     

An immunohistochemical and ultrastructural study on age-related astrocytic gliosis in the central nervous system of dogs.

The pattern of astrocytic gliosis (AG) was examined in 2-month-old to 18-year-old dogs using glial fibrillary acidic protein (GFAP) immunohistochemistry and electron microscopy. Coronal sections from various levels of the central nervous system (CNS) were stained with hematoxylin & eosin, Luxol Fast Blue, Nissl, and Bodian in addition to GFAP. A consistent pattern of age-related AG was observed in the dogs. The white matter, cortico-medullary junction, and subcortical nuclei in the cerebrum, central nuclei in the cerebellum, various nuclei in the brain stem, and grey matter of the spinal cord showed even and intense GFAP staining. AG was also prominent in the cerebral and cerebellar cortices and thalamus. Moderate AG was observed in the hippocampus and white matter of the cerebellum and spinal cord. Electron microscopy demonstrated increased number of profiles of degenerative neural components in the vicinity of hypertrophic astrocytes in the cerebral cortex of the aged dogs. Moderate to severe AG was consistently shown in the CNS of the aged dogs. In contrast, young normal dogs showed minimum amounts of GFAP-positive astrocytes in the CNS. These findings suggest that the observed AG in the CNS of the dogs is a morphological expression of aging.     

Morphometry of the thoracic spine in German shepherd dog: a computed tomographic study

Computed tomographic images of the thoracic spine of 13 German shepherd dogs were examined in order to determine the thoracic spine morphometry. Examinations were carried out in the transverse plane both intervertebral and mid-vertebral levels of the each thoracic vertebrae. The dorsoventral and interpedicular diameters of the spinal canal, the dorsoventral and transverse diameters of the vertebral body, the dorsoventral and transverse diameters of the spinal cord and also the cross-section area of the spinal canal were measured. The maximum values were found to be at the level of C7-T1. The shapes of the spinal canal and cord were circular in middle part, the shape became transverse oval in the cranial and caudal parts of the thoracic spine. The most significant correlation between the diameters was found to be in male dogs, except between dorsoventral diameters of the spinal canal and that of the vertebral body and between dorsoventral diameters of the spinal canal and transverse diameters of the vertebral body.