Equine laryngeal hemiplegia. Part II. An electron microscopic study of peripheral nerves

The recurrent laryngeal nerves were examined by electron microscopy in five control, four subclinical and four clinical laryngeal hemiplegic horses. In addition, the peroneal nerve was examined in two horses in the latter group. The distally distributed loss of large myelinated fibres in the left recurrent laryngeal nerve seen by light microscopy was confirmed. In addition, active axonal pathology was found to be more evident than indicated by light microscopic investigations. The onion bulb formations observed indicated the repetitive nature of the damaging influence to nerve fibres. Although the pathological changes were most obvious in the distal left recurrent laryngeal nerve, alterations similar in type and distribution were present in other areas of the left and right nerves, and in the distal hindlimb nerves. The observation of fibres with inappropriately thick myelin sheaths relative to their axonal calibre, was confirmed statistically by determining the regressions of axis cylinder perimeter against the number of myelin lamellae. In conclusion, the peripheral nerve pathology of equine laryngeal hemiplegia was demonstrated to be a distally distributed loss of myelinated fibres, with considerable active axonal damage, in conjunction with axonal atrophy. These features suggest that this disease may be classified as a distal axonopathy.     

Equine laryngeal hemiplegia. Part IV. Muscle pathology

This study confirmed that neurogenic muscle pathology exists in intrinsic laryngeal muscles supplied by the recurrent laryngeal nerves in horses subclinically and clinically affected with laryngeal hemiplegia. An important additional observation was the occurrence in three out of four laryngeal hemiplegic horses of neurogenic muscle changes in a hindlimb muscle, the extensor digitorum longus, a muscle supplied by another long peripheral nerve. This finding suggests that a polynenropathy exists in laryngeal hemiplegic horses, and supports the classification of this disease as a distal axonopathy. Comparison of the degree of pathology in the intrinsic laryngeal muscles and that of the recurrent laryngeal nerves innervating them, demonstrated a strong correlation between the extent of damage in the distal left recurrent laryngeal nerve and the overall degree of muscle pathology. The muscle damage in clinically affected horses is a reflection of the nerve damage present in the most distal portion of the recurrent laryngeal nerve. The more variable pathological changes found in proximal levels of the left and right recurrent laryngeal nerves probably reflects the ongoing nature of the pathological process affecting nerve fibres. The existence of a subclinically affected group of horses, the earliest involvement of an adductor, the left cricoarytenoideus lateralis muscle, and the presence of changes in the right intrinsic laryngeal muscles all confirmed the findings of previous workers.     

Pathological aspects of Australian Stringhalt.

Nine horses with clinical signs of Australian Stringhalt were killed and tissues collected for a detailed pathological study. Lesions were limited to peripheral nerves and muscles. The most severely affected nerves were the superficial and deep peroneal, distal tibial, plantar digital, volar and recurrent laryngeal nerve with changes characterised by a selective loss of large diameter myelinated fibres with various degrees of demyelination, fibrosis, Schwann cell proliferation and onion-bulb formation. A routine evaluation of the brain and spinal cord by light microscopy failed to reveal any consistent abnormalities. Morphometric analysis of deep peroneal and recurrent laryngeal nerves confirmed the reduced number of large diameter myelinated axons. Teased fibre preparations of these nerves did not show any abnormalities in internodal distance. The most severe muscle lesions were in the long and lateral digital extensors, cranial tibial, dorsal cricoarytenoid, gracilis and lateral deep digital flexor with extensive atrophy of fibres and diffuse fibrosis. Histochemical evaluation of the long digital extensor from 3 affected horses showed an abnormally wide distribution in fibre size and a reduction in type II fibres compared with controls. These lesions are consistent with a distal axonopathy leading to neurogenic muscle atrophy. The distribution of neuromuscular lesions in Australian Stringhalt may be explained by the susceptibility of longer, larger myelinated nerve fibres to injury, but the cause for this distal axonopathy remains unknown.     

Preferential denervation of the adductor muscles of the equine larynx. II: Nerve pathology

The terminal branches of the recurrent laryngeal nerve (RLN) of three normal ponies and six horses with sub-clinical laryngeal disease were examined qualitatively and quantitatively in an attempt to explain the preferential denervation of the laryngeal adductor muscles in the neuropathy of idiopathic laryngeal hemiplegia (ILH). The myelinated fibre spectra of all the motor nerve fibres in the left and right abductor and adductor branches of the RLN in three normal ponies were measured. The density of myelinated fibres was also calculated. There was no significant difference between the larger group of myelinated fibres in the adductor or abductor branches. In the six horses with laryngeal hemiparesis, however, there was a marked preferential loss of the medium/large size myelinated fibres in the left adductor branch, although nerve fibre densities were not significantly different. While no simple morphometrical feature was found to explain the selective muscle denervation, the greater loss of large diameter myelinated fibres in the adductor branches confirms the earlier observation of adductor muscle susceptibility in the neuropathy of ILH.     

Horner's syndrome and ipsilateral laryngeal hemiplegia in three cats

Horner's syndrome is described in three cats associated with wounds to the ventrolateral neck. In each case, ipsilateral laryngeal hemiplegia was observed on laryngoscopy. This finding provided strong evidence to support a diagnosis of second order Horner's syndrome due to disruption of the cervical sympathetic trunk, as motor fibres innervating laryngeal abductors also traverse the neck; both as descending fibres within the contiguous cervical vagus and as ascending fibres within the recurrent laryngeal nerve. Notably, the ability to vocalise was unimpaired in all cases and, in two cats, neck wounds were not apparent until the neck had been clipped and closely examined. These findings indicate that assessment of laryngeal function is of value when localising the site of the neural defect responsible for selected cases of second order Horner's syndrome.     

Stereological characteristics of the equine accessory nerve

Stereological techniques have been increasingly employed for assessment and characterization of neuromuscular diseases in humans and animals. As an adjunct to histopathology, morphometrical algorithms provide quantitative evidence of the peripheral nerve composition, thereby shedding light on its fibre characteristics and basic electrophysiological properties. In the horse, stereological investigations already have focussed on the recurrent laryngeal, deep peroneal and lateral palmar nerves (LPN). Of these, only the latter is suitable for taking biopsies in clinical settings, however, it does not contain any motor fibres and Ia-afferents. On account of its virtually mixed fibre qualities, most researchers today recommend the cervical branch of the equine accessory nerve (AN) for harvesting diagnostic samples. Thus, the present study was carried out to gain morphometrical proof of the AN composition and to obtain stereological base values in healthy individuals using state-of-the-art technology. All parameters were compared to the common peroneal nerve (CPN), known to harbour all myelinated fibre classes. As this second biopsy site is located farther distally to the neuro-axis, attention was paid to possible length-dependent features. Taken together, digital image analysis could be accurately applied on all AN samples. Stereology supported the histological and clinical evidence that the AN contains all myelinated fibre types. The huge range and scatter of fibre counts and density (3351-17,812/mm(2)) per fascicle were comparable to that measured in the equine common peroneal, deep peroneal, lateral palmar and recurrent laryngeal nerves. Similar to those, fibre diameter distribution was bimodal with slow Abeta- and Agamma-mechanoceptor afferents outnumbering large myelinated Aalpha-fibres by a factor of about 1.5. With a g-ratio at 0.55 +/- 0.001, the overall degree of myelination in the AN is highly consistent and insignificantly ranges between that of the equine common peroneal and LPNs. Apart from this subtle deviation, a statistically relevant difference between the more proximal AN and the distal CPN could not be documented. By obtaining morphometrical standard parameters and even more sophisticated distribution indices, stereology is a valuable tool for detection of subtle changes that are likely to escape from the investigators' eyes. The AN serves as a reliable source for advanced peripheral nerve research and should be accompanied by farther distal nerve probes for assessment of neuropathies that present with a proximodistal gradient.     

Morphometric study of the recurrent laryngeal nerve in young ‘normal’ horses

Quantitative measurements were made on cross-sectional preparations of the distal part of the recurrent laryngeal nerve (RLN) from nine young mixed-breed horses to establish reference values for the total number of myelinated fibres, mean fibre diameter and percentage of thickest fibres (over 9·5 gm) and to delineate diameter distribution curves. The total number of myelinated fibres, mean fibre diameter and percentage of thickest fibres for the left RLN were significantly lower than those of the right RLN (P<0–005). The distribution of fibres was unimodal. The fibre diameter ranged from 1 μm to 17 μm. Approximately, 95 per cent of fibres had a diameter larger than 5 μm.     

Bilateral arytenoid cartilage paralysis after inhalation anesthesia in a horse

An 8-year-old Quarter Horse gelding was anesthetized for surgical exploration and debridement of a chronic draining wound in the intermandibular space. Anesthesia was without complication other than persistently low PaO2. Severe airway obstruction was evident immediately after extubation, requiring tracheostomy. Endoscopic diagnosis was bilateral arytenoid paralysis, which gradually resolved over the next 7 days. Compression, trauma, or tension of the recurrent laryngeal nerves are the postulated causes of idiopathic laryngeal hemiplegia in horses. The extremely extended position of the head and neck during anesthesia, perhaps compounded by low arterial oxygen content, may have resulted in a hypoxemic insult to the recurrent laryngeal nerves and caused bilateral arytenoid paralysis in this horse.     

Peripheral nerve pathology in two rottweilers with neuronal vacuolation and spinocerebellar degeneration

Neuronal vacuolation and spinocerebellar degeneration in young Rottweiler dogs is a neurodegenerative condition characterized by neuronal vacuolation of several nuclei in the central nervous system and degeneration of the spinal cord white matter. Here, we describe the morphologic and ultrastructural findings in laryngeal muscles and peripheral nerves of a 16-week-old female and a 32-week-old female Rottweiler dog affected by progressive ataxia and tetraparesis associated with laryngeal paralysis. Lesions were characterized by neurogenic muscle atrophy of the intrinsic laryngeal muscles, and a loss of large myelinated fibers in the recurrent laryngeal nerve, accompanied by demyelinating/remyelinating features affecting the small myelinated fibers. No significant changes were detected in the cranial laryngeal, vagus, phrenic, ulnar, or peroneal nerves. These findings were indicative of a selective distal neuropathy of the recurrent laryngeal nerve with early severe axonal degeneration, mainly of the large myelinated fibers.     

Bilateral laryngeal paralysis in the horse

Four two-year-old Thoroughbreds suffered an acute gastrointestinal illness shortly after dosing with mineral oil which was thought to have been contaminated with an organophosphate compound. Three weeks later all four were noted to be dyspnoeic and endoscopic examination showed that they had developed bilateral laryngeal paralysis. Two of the horses died during severe bouts of dyspnoea six and eight months later and the third was killed shortly thereafter. Examination of the left and right recurrent laryngeal nerves from these horses showed a severe loss of myelinated fibres distally, especially in the left nerve. A similar but less severe lesion was seen in other long peripheral nerves, including the phrenic and digital nerves of the third horse. The spinal cord in two horses showed evidence of mild axonal degeneration which was not related to a particular tract or location. The fourth horse had bilateral laryngeal paralysis two years later. The acute clinical signs and delayed neurological sequelae seen in these horses were strongly suggestive of accidental organophosphate toxicity.     

Attempts to restore abduction of the paralyzed equine arytenoid cartilage. III. Nerve anastomosis.

The purpose of this project was to attempt restoration of abduction of a recently denervated left dorsal cricoarytenoid muscle in the horse by anastomosing the first cervical nerve to the abductor branch of the left recurrent laryngeal nerve. Ten horses were used in the study. In six horses the left recurrent laryngeal nerve was transected and ligated while the ventral branch of the left first cervical nerve was anastomosed to the abductor branch of the left recurrent laryngeal nerve. The remaining four horses also had the left recurrent laryngeal nerve transected and ligated but had no nerve anastomosis performed. Each horse was evaluated preoperatively, and at one week, three and six months after surgery, by endoscopy and determination of upper airway resistance. The endoscopy was performed with the horses breathing room air and while breathing 10% carbon dioxide. All ten horses showed endoscopic signs of complete laryngeal hemiplegia immediately postoperatively. Starting at three months postoperatively clonic movements of the left arytenoid cartilage were observed in four of the six reinnervated horses but not in the sham operated horses. At the sixth postoperative month five reinnervated horses had clonic movements of the left arytenoid cartilage. The comparison of upper airway resistance measurements before surgery and at one week, three and six months after surgery showed no significant differences in either control or experimental horses. Following euthanasia at six months postoperatively, the left and right dorsal crioarytenoid muscles were compared for evidence of reinnervation. No significant difference in weight was noted in the reinnervated horses but the left dorsal cricoarytenoid muscle weighed less than the control horses.     

Experimental crush of the equine recurrent laryngeal nerve: a study of normal and aberrant reinnervation

Experimental reinnervation of the equine larynx in healthy ponies was studied after the recurrent laryngeal nerve was crushed, using endoscopic, electromyographic, and microscopic techniques. In 12 ponies, the recurrent laryngeal nerve was crushed unilaterally in the midcervical area. All showed postoperative paralysis of the larynx on the operated side. In 8 ponies, recovery of movement of the vocal folds occurred at different times (2.5 to 8 months) after surgical operation was done. These movements, which were often abnormal, included trembling and asynchronism. In 2 of these ponies, complete return of normal vocal fold function occurred. Return of function was due to reinnervation, as determined by electromyographic examination of the laryngeal muscles and microscopic evaluation of these muscles and the recurrent laryngeal nerves. In the muscles, there was evidence of fiber-type grouping, and the nerves showed regenerative activity. In 3 ponies, there was evidence of aberrant reinnervation, with adductor or abductor muscles discharging during an inappropriate phase of the respiratory cycle. Further evidence that reinnervation occurred in the larynges of these ponies was determined in microscopic sections of the recurrent laryngeal nerves and muscles, which showed regenerative activity and muscle fiber-type grouping, respectively. The endoscopic abnormalities in these ponies were compared with those seen in horses with spontaneous subclinical laryngeal paralysis. However, the possibility that the abnormal endoscopic findings in this latter group of horses are due to aberrant reinnervation is thought to be unlikely.     

Laryngeal Paralysis-Polyneuropathy Complex in Young Rottweilers

Five Rottweiler puppies from 3 unrelated litters developed inspiratory stridor at 11–13 weeks of age. Physical examination disclosed tetraparesis in all dogs, and bilateral lenticular cataracts in 4 dogs. Laryngeal examination under light anesthesia showed laryngeal paralysis in all dogs. Electrodiagnostic testing revealed denervation potentials in the distal appendicular muscles of 4 dogs tested and in the intrinsic laryngeal muscles of 2 dogs tested. Motor nerve conduction velocity was slightly low in 1 dog. Neurogenic muscular atrophy was found in distal appendicular muscles (n = 3) and intrinsic laryngeal muscles (n = 2), and degenerative changes were found in peripheral nerves (n = 3) and recurrent laryngeal nerves (n = 2). No abnormalities were detected in the spinal cord, spinal nerve roots, or ganglia of 3 dogs autopsied. The clinical, electrophysiologic, and histopathologic findings support a diagnosis of polyneuropathy and resemble the finding reported in young Dalmatians. Young dogs with laryngeal paralysis should be evaluated neurologically to rule out a more generalized polyneuropathy. The condition is suspected to be hereditary in nature and the prognosis is poor.     

Interpretation of laryngeal function tests in the horse

Idiopathic left-sided laryngeal paralysis was present in 14 of 169 horses on a thoroughbred horse farm (8.3 per cent). In nine animals, it was evident only after exercise and arytenoid abduction and adduction were normal at rest. Asynchronous movement of the arytenoid cartilages was observed in 94 horses at rest (55.6 per cent), 86 of which were considered to be normal after exercise. Conversely, synchronous movement of the arytenoids was noted when at rest in six of the 14 animals diagnosed as having laryngeal hemiplegia after exercise. An abnormal inspiratory noise during exercise was detectable in 11 of these 14 horses, but not in the remainder. An abnormal noise on inspiration was also produced by nine horses in which laryngeal hemiplegia was not diagnosed.     

Examination of the laryngeal musculature and recurrent laryngeal nerves of zebra (Equus burchelli) for evidence of idiopathic laryngeal hemiplegia

AIMS: This study was undertaken in order to determine whether wild Burchell's Zebra (Equus burchelli) are affected by Idiopathic Laryngeal Hemiplegia (ILH). ILH has a high prevalence in the domestic horse and a negative control population has not been identified. METHODS: The work was conducted in two phases on two large farms in central Kenya during routine culls. The larynges of 42 zebra were removed and the left and right intrinsic laryngeal adductor and abductor muscles and recurrent laryngeal nerves were isolated. The muscles were weighed and biopsy samples of left and right recurrent laryngeal nerve and cricoarytenoideus lateralis muscles from 16 animals were taken for histological examination. RESULTS: Left adductor muscles were observed to be significantly lighter than right adductor muscles (p = 0.001). There was no significant difference between the weights of left and right laryngeal abductors (p = 0.260). There was no light microscopic evidence of pathological changes consistent with ILH in any of the muscle or nerve samples. CONCLUSIONS: Zebra do not appear to suffer from ILH and equids unaffected by ILH can have significantly lighter muscles on the left of the larynx. CLINICAL RELEVANCE: Mild asymmetries of the larynx on palpation may not be a reliable indicator of ILH. Zebra may be a potentially useful negative control animal for observational ILH studies.     

Equine laryngeal hemiplegia. Part V. Central nervous system pathology

Evidence of long central nerve fibre degeneration (axonal spheroids) in the lateral cuneate nuclei was found in all eight Thoroughbreds affected clinically and subclinically with equine laryngeal hemiplegia, but in only one of six control animals. It was considered that these spheroids may signify a central nervous component of the disease process of laryngeal hemiplegia although until further investigations are performed no firm conclusions regarding the relationship of these findings with laryngeal hemiplegia could be made. Examination of the left and right nucleus ambiguus of clinical and subclinical laryngeal hemiplegic horses revealed no pathological alterations.     

Use of the forced oscillating airflow technique to measure the resistance of the equine upper airway: effects of laryngoventriculectomy and laryngoplasty

Investigations were carried out to determine whether measurements of total respiratory resistance (TRR) made in resting animals could detect changes due to laryngeal hemiplegia. Control values of TRR were obtained in eight ponies and in six of these the measurements were repeated after division of the left recurrent laryngeal nerve in the mid-cervical region; two were retained as controls. A further set of measurements were made before two of the operated animals were subjected to left laryngoventriculectomy (Hobday operation). A laryngoplasty ('tie-back' operation) was performed on another two animals. Measurements of TRR were made at three-monthly intervals on all eight ponies for the next two years. Endoscopic examination of the larynx was carried out at the time of the measurements. All ponies showed a marked, but regular, variation in TRR over the two-year observation period. Nerve section was shown to be associated with an increase in TRR in five ponies. The Hobday operation decreased TRR in one animal and had no marked effect in the second. The tie-back operation was associated with a decreased TRR in one pony and an increase in the second. Autopsies were carried out on all six operated ponies and measurements of TRR and lower respiratory resistance (LRR) were made immediately before the animals were killed. Evidence of regeneration of the recurrent laryngeal nerve was seen in one of the ponies which underwent the Hobday operation. Histological evidence of regeneration was found in another pony.(ABSTRACT TRUNCATED AT 250 WORDS)     

Morphologic and morphometric studies of the vagus and recurrent laryngeal nerves in clinically normal adult dogs

Morphologic and morphometric studies were performed on the vagus nerve and its major branch, the recurrent laryngeal nerve (RLN): (1) to determine normal histologic data in myelinated fibers of clinically normal young adult dogs, (2) to establish reference values for mean fiber diameter in the vagus nerve and in the proximal and distal portions of the RLN, and (3) to delineate relative frequency distribution curves for each nerve. Few degenerative changes were observed in single teased-nerve fiber preparations. There was no statistical difference between left and right sides of the vagus nerve or between the proximal and distal portions of the RLN (right and left sides). In contrast to the unimodal distribution of fibers in the vagus nerve and the proximal portion of the RLN, the distribution of fibers in the distal portion of the RLN was bimodal. Mean (+/- SD) fiber diameters of the vagus nerve and in the proximal portion of the RLN (3.02 +/- 1.44 microns and 3.63 +/- 1.49 microns, respectively) were not significantly different, despite a shift to large-diameter fibers in the latter. However, mean fiber diameters of distal and proximal portions of the RLN (5.56 +/- 1.88 microns and 3.63 +/- 1.49 microns, respectively) were significantly (P less than 0.001) different. Approximately 86% of fibers in the vagus nerve and 76% of fibers in the proximal portion of the RLN, had a diameter less than 5 microns, and about 70% of fibers in the distal portion of the RLN had a diameter greater than 5 microns.(ABSTRACT TRUNCATED AT 250 WORDS)     

Preferential denervation of the adductor muscles of the equine larynx. I: Muscle pathology

The laryngeal muscles of 18 horses were examined histologically. The neurogenic changes found in each muscle were scored by four reviewers and the results evaluated statistically. Fifteen of these horses had endoscopic evidence of abnormal laryngeal function, three of which were defined as having adductor paralysis. Measurement of muscle fibre area in two horses with idiopathic laryngeal hemiplegia (ILH) was performed. In the quantitative study of neurogenic change, the adductor muscles were more significantly affected than the abductor muscle. This was also true in the clinical cases of ILH where measurement of muscle fibre area demonstrated that the lateral cricoarytenoid (adductor) muscles showed a wider range of pathological changes than the dorsal cricoarytenoid muscle (abductor). Those horses with the most severe muscle pathology also had the most abnormal endoscopic findings. The propensity for denervation of the adductor muscles should provide clues as to the pathogenesis and natural history of horses with sub-clinical laryngeal disease and ILH.