Histologic evaluation of nerve muscle pedicle graft used as a treatment for left laryngeal hemiplegia in standardbreds.

A nerve muscle pedicle (NMP) graft was placed in the cricoarytenoideus dorsalis (CAD) muscle of 6 horses with induced left laryngeal hemiplegia. The NMP graft was created by use of the first cervical nerve and omohyoideus muscle. In 1 horse (control), the first cervical nerve was transected after placement of the NMP graft. One year after the surgical procedure, horses were examined endoscopically and then anesthetized. While the larynx was observed endoscopically, the first cervical nerve was stimulated. Horses were subsequently euthanatized, and the larynx was harvested. Prior to anesthesia, the endoscopic appearance of the larynx of all horses was typical of laryngeal hemiplegia. During anesthesia, stimulation of the first cervical nerve produced vigorous abduction of the left arytenoid in principal horses but not in the control horse. The right cricoarytenoideus lateralis and CAD muscles were grossly and histologically normal. Also, the left cricoarytenoideus lateralis was atrophic in all horses as was the left CAD muscle of the control horse. In contrast, the left CAD muscle harvested from principal horses had evidence of reinnervation with type 1 or type 2 fiber grouping. One year after the NMP graft procedure, horses with left laryngeal hemiplegia had reinnervation of the left CAD muscle. In another study, reinnervation was sufficient to allow normal laryngeal function during exercise. Combined, these data suggest that the NMP graft procedure is a viable technique for the treatment of left laryngeal hemiplegia in horses.     

Attempts to restore abduction of the paralyzed equine arytenoid cartilage. I. Nerve-muscle pedicle transplants.

The purpose of this project was to adapt a surgical technique from humans and dogs to horses in which a portion of an accessory muscle of respiration and its nerve supply is transplanted to a denervated dorsal cricoarytenoid muscle. Anatomical dissections in seven horses revealed two possible donor nerve-pedicle grafts: the omohyoid and the sternothyrohyoid, both innervated by a branch of the first and second cervical nerves. Histochemical evaluations in two ponies of the dorsal cricoarytenoid, omohyoid and sternothyrohyoid muscles revealed similar proportions of fiber types 1 and 2 in all three muscles. Electromyographic studies in these two ponies revealed that the omohyoid and sternothyrohyoid muscles contract synchronously with respirations during forced inspiration under general anesthesia. Based on surgical ease of access, a 1 cm2 portion of the omohyoid muscle at the point of penetration of the second cervical nerve was used as a nerve-muscle pedicle graft in an attempt to reinnervate the left dorsal cricoarytenoid muscle in four ponies. These four ponies (as well as three others which served as controls) had previously undergone left recurrent laryngeal nerve transection. All seven ponies endoscopically showed signs of complete left laryngeal hemiplegia immediately postoperatively. Animals were monitored endoscopically for 30 weeks after surgery. The three control ponies showed no abduction of the arytenoid cartilage. In addition, in these three ponies, histological and histochemical expected changes of muscle fiber atrophy and fibrosis were present in the dorsal cricoarytenoid muscle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Equine laryngeal hemiplegia. Part I. A light microscopic study of peripheral nerves

This light microscopic investigation of 15 Thoroughbred horses provided substantial evidence for the classification of equine laryngeal hemiplegia as a distal axonopathy. Morphologic and morphometric examinations were performed on resin embedded recurrent laryngeal nerves from control, subclinical and clinical laryngeal hemiplegic animals. In the latter group of animals some distal hindlimb nerves were also examined. A distally graded loss of myelinated fibres selectively affecting those of large diameter was demonstrated in both left and right recurrent laryngeal nerves. Morphologic evidence of similar pathological changes in long hindlimb nerves was also present. An explanation for the early involvement of the cricoarytenoideus lateralis muscle in the course of laryngeal hemiplegia, was offered by the demonstration of more large diameter fibres in the branch of the recurrent laryngeal nerve innervating it, thus making it more susceptible to the disease process.     

Laryngeal paralysis: a study of 375 cases in a mixed-breed population of horses

Referred cases (n = 375) of laryngeal paralysis (1985-1998) from a mixed-breed equine population included 351 (94%) cases of recurrent laryngeal neuropathy (RLN) (idiopathic laryngeal hemiplegia) and 24 cases (6%) of laryngeal paralysis from causes other than RLN. Laryngeal movements were classified endoscopically into one of 6 grades, in contrast to the usual 4 grades. The RLN cases had a median grade 4 laryngeal paralysis, of which 96% were left-sided, 2% right-sided and 2% bilaterally affected. RLN cases included 204 (58%) Thoroughbred, 96 (27%) Thoroughbred-cross, 23 (7%) draught, 16 (5%) Warmbloods and 10 (3%) other breeds, including only 4 (1%) ponies. The median age of RLN cases at referral was 6 years (range 2-12) and their median height was 170.2 cm. The work of RLN horses included National Hunt racing (42%), flat racing (1%), hunting (19%), eventing (16%) and miscellaneous work (22%). Reported presenting signs in RLN-affected horses included abnormal exercise-related respiratory sounds in 90% and reduced exercise tolerance in only 64%. However, many horses were referred before their exercise tolerance could be fully assessed. Forty percent of the RLN cases had intercurrent disorders, including 10% with additional upper respiratory and 7% with lower respiratory tract diseases. The 24 nonidiopathic RLN cases included 12 with bilateral laryngeal paralysis, 11 (92%) of which were ponies. Bilateral laryngeal paralysis occurred with hepatic encephalopathy in 7 cases and following general anaesthesia in 2 cases. The 12 cases of acquired unilateral laryngeal paralysis included 7 caused by guttural pouch mycosis.     

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 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.     

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.     

A study of an electrodiagnostic technique for the evaluation of equine recurrent laryngeal neuropathy

The electrodiagnostic measurement of the thoracolaryngeal reflex (TLR) ('slap test') latency was compared to 5 other diagnostic techniques used for evaluation of laryngeal function, namely laryngeal muscle palpation, resting and immediately postexercise endoscopic examinations, and palpable and endoscopic responses to the TLR. Compared to resting endoscopy, the electrodiagnostic measurement of TLR latency was not found to be an accurate test for the evaluation of recurrent laryngeal neuropathy (RLN), nor was laryngeal muscle palpation or the endoscopic response to the TLR. Twenty-five (71%) of 35 Clydesdale horses examined were affected by RLN; 16 (46%) had mild, 5 (14%) moderate, 3 (9%) severe hemiparesis and 1 (3%) had total laryngeal paralysis. Within these 35 horses, physical traits such as height or neck length did not correlate with the incidence or severity of RLN. Ten control ponies showed no evidence of abnormal laryngeal function.     

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.     

Myosin heavy chain composition in normal and atrophic equine laryngeal muscle

The myosin heavy chain (MHC) composition of a given muscle determines the contractile properties and, therefore, the fiber type distribution of the muscle. MHC isoform expression in the laryngeal muscle is modulated by neural input and function, and it represents the cellular level changes that occur with denervation and reinnervation of skeletal muscle. The objective of this study was to evaluate the pattern of MHC isoform expression in laryngeal muscle harvested from normal cadavers and cadavers with naturally occurring left laryngeal hemiplegia secondary to recurrent laryngeal neuropathy. Left and right thyroarytenoideus (TA) and cricoarytenoideus dorsalis (CAD) were obtained from 7 horses affected with left-sided intrinsic laryngeal muscle atrophy and from 2 normal horses. Frozen sections were evaluated histologically for degree of atrophy and fiber type composition. MHC isoform expression was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of muscle protein. Histologic atrophy was seen in all atrophic muscles and some right-sided muscles of 3 affected horses, as well as the left TA of 1 normal horse. Fiber type grouping or loss of type I muscle fibers was observed in the left-sided laryngeal muscles in all but 1 affected horse, as well as in the right muscles of 2 affected horses, and the left TA of 1 normal horse. SDS-PAGE showed 2 bands corresponding to the type I and type IIB myosin isoforms in the CAD and TA of the 2 normal horses. Affected horses demonstrated a trend toward increased expression of the type IIB isoform and decreased expression of the type I isoform in atrophic muscles. This study confirmed the presence of histologic abnormalities in grossly normal equine laryngeal muscle, and it demonstrated an increased expression of type IIB MHC with a concurrent decreased expression of type I MHC in affected muscles. Evaluation of muscle fiber changes at the cellular level under denervated and reinnervated conditions may aid in assessing future strategies for reinnervation or regeneration of atrophic laryngeal muscle.     

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.     

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.     

DIAGNOSTIC SENSITIVITY OF SUBJECTIVE AND QUANTITATIVE LARYNGEAL ULTRASONOGRAPHY FOR RECURRENT LARYNGEAL NEUROPATHY IN HORSES

Recurrent laryngeal neuropathy (RLN) is the most common cause of laryngeal hemiplegia in horses and causes neurogenic atrophy of the intrinsic laryngeal muscles, including the cricoarytenoideus lateralis muscle. Recurrent laryngeal neuropathy results in paresis to paralysis of the vocal fold and arytenoid cartilage, which limits performance through respiratory compromise. Ultrasound has previously been reported to be a useful diagnostic technique in horses with RLN. In this report, the diagnostic sensitivity of subjective and quantitative laryngeal ultrasonography was evaluated in 154 horses presented for poor performance due to suspected upper airway disease. Ultrasonographic parameters recorded were: cricoarytenoideus lateralis echogenicity (subjective and quantitative), cricoarytenoideus lateralis thickness, vocal fold movement, and arytenoid cartilage movement. Ultrasonographic parameters were then compared with laryngeal grades based on resting and exercising upper airway endoscopy. Subjectively increased left cricoarytenoideus lateralis echogenicity yielded a sensitivity of 94.59% and specificity of 94.54% for detecting RLN, based on the reference standard of exercising laryngeal endoscopy. Quantitative left cricoarytenoideus lateralis echogenicity values differed among resting laryngeal grades I–IV. Findings from this study support previously published findings and the utility of subjective and quantitative laryngeal ultrasound as diagnostic tools for horses with poor performance.     

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.     

Histological and ultrastructural evidence that recurrent laryngeal neuropathy is a bilateral mononeuropathy limited to recurrent laryngeal nerves

Reasons for performing study: Recurrent laryngeal neuropathy (RLN) is a common and debilitating peripheral nerve disease of horses, but it remains unclear if this disease is a mono‐ or polyneuropathy. An understanding of the distribution of the neuropathological lesions in RLN affected horses is fundamental to studying the aetiology of this very significant disease of tall horses. Objective: To determine whether RLN should be classified as a mono‐ or polyneuropathy. Methods: Multiple long peripheral nerves and their innervated muscles were examined systematically in 3 clinically affected RLN horses Results: Severe lesions were evident in the left as well as right recurrent laryngeal nerves in all horses, both distally and, in one case, also proximally. No primary axonal lesions were evident in other nerves nor were changes found in their innervated muscles. Conclusions: RLN is not a polyneuropathy but should be classified as a bilateral mononeuropathy. Potential relevance: Genetic and local factors specifically affecting the recurrent laryngeal nerves in RLN‐affected horses should now be investigated further.     

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.     

Partial Laryngectomy in the Dog I. Healing and Swallowing Function in Normal Dogs

Laryngeal paralysis was created in normal dogs by sectioning the recurrent laryngeal nerves. The dogs were treated with bilateral vocal fold resection and unilateral partial arytenoidectomy, and sacrificed from 6 hours to 28 days following surgery. Swallowing abnormalities were not found in any of the dogs. Healing by granulation-epithelialization was complete by 14 days.     

Innervation of the equine intrinsic laryngeal muscles

The nerve supply to the intrinsic laryngeal muscles of the horse was studied by gross dissection and by electromyography which was carried out before, during and after section of various intralaryngeal nerve branches. The anatomical relationships and passage of the laryngeal nerves throughout the larynx were defined. Unlike the dog and man there was no evidence of the passage of motor nerve fibres from one side of the larynx to the other.     

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.     

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.