Electrophysiologic Study of a Method of Euthanasia Using Intrathecal Lidocaine Hydrochloride Administered during Intravenous Anesthesia in Horses

Background

An intravenous (IV) overdose of pentobarbital sodium is the most commonly used method of euthanasia in veterinary medicine. However, this compound is not available in many countries or rural areas resulting in usage of alternative methods such as intrathecal lidocaine administration after IV anesthesia. Its safety and efficacy as a method of euthanasia have not been investigated in the horse.

Hypothesis/Objectives

To investigate changes in mean arterial blood pressure and electrical activity of the cerebral cortex, brainstem, and heart during intrathecal administration of lidocaine. Our hypothesis was that intrathecal lidocaine affects the cerebral cortex and brainstem before affecting cardiovascular function.

Animals

Eleven horses requiring euthanasia for medical reasons.

Methods

Prospective observational study. Horses were anesthetized with xylazine, midazolam, and ketamine; and instrumented for recording of electroencephalogram (EEG), electrooculogram (EOG), brainstem auditory evoked response (BAER), and electrocardiogram (ECG). Physical and neurological (brainstem reflexes) variables were monitored. Mean arterial blood pressure was recorded throughout the study.

Results

Loss of cerebro‐cortical electrical activity occurred up to 226 seconds after the end of the infusion of lidocaine solution. Cessation of brainstem function as evidenced by a lack of brainstem reflexes and disappearance of BAER occurred subsequently. Undetectable heart sounds, nonpalpable arterial pulse, and extremely low mean arterial blood pressure supported cardiac death; a recordable ECG was the last variable to disappear after the infusion (300–1,279 seconds).

Conclusions and Clinical Importance

Intrathecal administration of lidocaine is an effective alternative method of euthanasia in anesthetized horses, during which brain death occurs before cardiac death.     

Brainstem Auditory Evoked Responses in Foals: Reference Values,Effect of Age,Rate of Acoustic Stimulation,and Neurologic Deficits

Background

Age and rate of acoustic stimulation affect peak latencies in brainstem auditory evoked responses (BAER) in humans. Those effects are unknown in foals.

Hypothesis/Objectives

Our goals were to (1) establish reference values for BAER in foals by using 3 different stimulation protocols, (2) evaluate the effects of age and stimulation frequencies on BAER tracing in foals up to 6 months old, and (3) compare the data with BAER obtained from foals with central nervous system (CNS) disorders.

Animals

Thirty‐nine neurologically normal foals and 16 foals with neurologic diseases.

Methods

Prospective observational clinical study. BAER recorded by using 3 protocols of stimulation (11.33 repetitions per second [Hz]/70 decibel normal hearing level [dBNHL]; 11.33 Hz/90 dBNHL; 90 Hz/70 dBNHL).

Results

No effect of age was observed in normal foals (P > .005). No significant difference was observed for latencies and interpeak latencies (IPL) when comparing foals with neurologic diseases and normal foals (P > .05), but 78.6% of foals with neurologic diseases had an asymmetry in their tracing, reflecting a difference in conduction time between the left and right side of the brainstem. Increasing the stimulation rate did not improve detection of CNS disorders.

Conclusions and Clinical Importance

We propose BAER reference values for foals up to 6 months of age by using 3 protocols. Most foals with neurologic deficits had abnormal BAER tracing.     

Prevalence of unilateral and bilateral deafness in border collies and association with phenotype

BACKGROUND: Congenital sensorineural deafness (CSD) occurs in Border Collies, but its prevalence and inheritance are unknown. This study estimated the prevalence of CSD in Border Collies and investigated its association with phenotypic attributes linked to the merle gene, including coat pigmentation and iris color. HYPOTHESIS: Deafness in Border Collies is associated with pigmentation patterns linked to the merle gene. ANIMALS: A total of 2597 Border Collies from the United Kingdom. METHODS: A retrospective study of Border Collies tested, during 1994-2002, by using brainstem auditory evoked responses. Associations between deafness and phenotypic attributes were assessed by using generalized logistic regression. RESULTS: The prevalence of CSD in puppies was estimated as 2.8%. The corresponding rates of unilateral and bilateral CSD were 2.3 and 0.5%, respectively. Adjustment for clustering of hearing status by litter reduced the overall prevalence estimate to 1.6%. There was no association between CSD and sex (P = .2). Deaf Border Collies had higher rates of merle coat pigmentation, blue iris pigment, and excess white on the head than normal hearing Border Collies (all P < .001). The odds of deafness were increased by a factor of 14 for Border Collies with deaf dams, relative to the odds for dogs with normal dams (P = .007), after adjustment for phenotypic attributes. CONCLUSIONS AND CLINICAL IMPORTANCE: Associations between CSD and pigmentation patterns linked to the merle gene were demonstrated for Border Collies. Evidence for an inherited component to CSD in Border Collies supports selective breeding from only tested and normal parents to reduce the prevalence of this disease.     

Malignant Hyperthermia Associated with Ryanodine Receptor 1 (C7360G) Mutation in Quarter Horses

Background: Anesthetic‐induced malignant hyperthermia (MH) has been documented in Quarter Horses with a single point mutation in the ryanodine receptor 1 gene (RyR1) at nucleotide C7360G, generating a R2454G amino acid substitution. However, there have been no reports of nonanesthetic manifestations of MH in horses with the C7360G mutation. Objective: To describe clinical manifestations of Quarter Horses with the C7360G mutation. Animals: Eleven Quarter Horses with the RyR1 C7360G mutation. Methods: This prospective study included horses with suspected MH, undetermined etiology of sudden death, death within hours of onset of rhabdomyolysis, muscle rigidity, stiffness, intermittent sweating, and persistent increases in serum muscle enzyme activities. Whole blood in EDTA and skeletal muscle were processed for genetic and histochemical analysis. Medical records and pedigrees were collected when available. Results: Both anesthetic‐ and non–anesthetic‐associated myopathic manifestations of MH occurred in halter Quarter Horses with mutation of RyR1. The disease is inherited as an autosomal dominant trait. Clinical and laboratory abnormalities were similar in both forms. Rhabdomyolysis was a common finding in both groups of horses. Skeletal muscle histochemical findings were nonspecific and compatible with a noninflammatory myopathic process. Conclusions and Clinical Importance: MH is a potentially fatal disease of Quarter Horses that could be triggered by halogenated anesthetics and other nonanesthetic factors that may include exercise, stress, breeding, illnesses, and concurrent myopathies.     

Audiograms estimated from brainstem tone-evoked potentials in dogs from 10 days to 1.5 months of age

The objective of this study was to build audiograms from thresholds of brainstem tone-evoked potentials in dogs and to evaluate age-related change of the audiogram in puppies. Results were obtained from 9 Beagle puppies 10-47 days of age. Vertex to mastoid brainstem auditory-evoked potentials in response to 5.1-millisecond Hanning-gated sine waves with frequencies octave-spaced from 0.5 to 32 kHz were recorded. Three dogs were examined at 10, 13, 19, 25, and 45 days. Four other dogs were examined at 16 days. Data from 7 dogs between 42 and 47 days of age were pooled to obtain audiogram reference values in 1.5-month-old puppies. The best auditory threshold lowered from above 60 dB sound pressure level (SPL) to values close to 0 dB SPL between 13 and 25 days of age and then stabilized. The audible frequency range widened, including 32 kHz in all tested dogs from the 19th day. In the 7 1.5-month-old puppies, the mean auditory threshold decreased by 11 dB per octave from 0.5 to 2 kHz. The auditory threshold was lowest and held the same value from 2 to 8 kHz. The mean auditory threshold increased by 20 dB per octave from 8 to 32 kHz. Near threshold, click-evoked potentials test only a small part of the audible frequency range in dogs. Use of tone-evoked potentials may become a powerful tool in investigating dogs with possible partial hearing loss, including during the auditory system maturation period.     

Antiarrhythmic and Electrophysiologic Effects of Flecainide on Acutely Induced Atrial Fibrillation in Healthy Horses

Background

Only few pharmacologic compounds have been validated for treatment of atrial fibrillation (AF) in horses. Studies investigating the utility and safety of flecainide to treat AF in horses have produced conflicting results, and the antiarrhythmic mechanisms of flecainide are not fully understood.

Objectives

To study the potential of flecainide to terminate acutely induced AF of short duration (≥15 minutes), to examine flecainide‐induced changes in AF duration and AF vulnerability, and to investigate the in vivo effects of flecainide on right atrial effective refractory period, AF cycle length, and ventricular depolarization and repolarization.

Animals

Nine Standardbred horses. Eight received flecainide, 3 were used as time‐matched controls, 2 of which also received flecainide.

Methods

Prospective study. The antiarrhythmic and electrophysiologic effects of flecainide were based on 5 parameters: ability to terminate acute pacing‐induced AF (≥15 minutes), and drug‐induced changes in atrial effective refractory period, AF duration, AF vulnerability, and ventricular depolarization and repolarization times. Parameters were assessed at baseline and after flecainide by programmed electrical stimulation methods.

Results

Flecainide terminated all acutely induced AF episodes (n = 7); (AF duration, 21 ± 5 minutes) and significantly decreased the AF duration, but neither altered atrial effective refractory period nor AF vulnerability significantly. Ventricular repolarization time was prolonged between 8 and 20 minutes after initiation of flecainide infusion, but no ventricular arrhythmias were detected.

Conclusions and Clinical Importance

Flecainide had clear antiarrhythmic properties in terminating acute pacing‐induced AF, but showed no protective properties against immediate reinduction of AF. Flecainide caused temporary prolongation in the ventricular repolarization, which may be a proarrhythmic effect.     

Effects of heparin, citrate, and EDTA on plasma biochemistry of sheep: comparison with serum

This study was carried out to evaluate the features of neurological dysfunction in experimentally-induced bovine spongiform encephalopathy (BSE)-infected cattle using brainstem auditory evoked potentials (BAEP). The progressive prolongation of peak latency of waves III and V was observed right-and-left bilaterally at the onset of neurological symptoms. The peak latency of wave V and the I-V interpeak latency (IPL) in BSE cattle 22 and 24 months after intracerebral inoculation were significantly (P < 0.05) prolonged compared with the control cattle. In addition, the amplitude of the BAEP waves of the BSE cattle were low compared with the control cattle. Hearing loss occurred in the BSE cattle that showed advanced neurological symptoms such as tremor. It is thought that this BAEP data reflects a functional disorder in the central auditory nerve pathways characteristic of experimentally-induced BSE.     

Peritoneal d-Dimer Concentration for Assessing Peritoneal Fibrinolytic Activity in Horses with Colic

Background: Plasma d -dimer concentration is a useful marker to assess systemic coagulation and fibrinolytic activities in humans, dogs, and horses. Peritoneal fibrinolytic activity increases in horses with colic, especially in horses with endotoxin in the peritoneal fluid.
Hypothesis/Objectives: Peritoneal d -dimer concentration can be used to assess peritoneal fibrinolytic activity in horses with severe gastrointestinal (GI) disorders and altered peritoneal fluid.
Animals: Two hundred and twenty-one colic horses and 15 control horses.
Methods: Prospective observational clinical study. Blood and peritoneal fluid were collected on admission. Horses were grouped according to diagnosis, peritoneal fluid analysis, and outcome. Peritoneal d -dimer concentration was determined, together with peritoneal tissue-plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1) activities. Plasma d -dimer concentration also was measured.
Results: Peritoneal d -dimer concentration was significantly higher in all colic groups compared with controls, and in horses with enteritis, peritonitis, and ischemic disorders compared with horses with large intestinal obstructions. Peritoneal d -dimer concentration was significantly higher in horses with altered peritoneal fluid (modified transudate and exudate) compared with horses with normal peritoneal fluid analysis. Plasma d -dimer concentration also was significantly higher in the peritonitis group, and in horses with altered peritoneal fluid analysis. Peritoneal and plasma d -dimer concentrations also were significantly higher in nonsurvivors. Peritoneal d -dimer concentration was significantly correlated with decreased peritoneal t-PA activity and increased peritoneal PAI-1 activity.
Conclusions and Clinical Importance: Peritoneal d -dimer concentration is markedly higher in severe GI disorders, and it can be used to assess peritoneal fibrinolytic activity in horses with colic.     

Glucocorticoid Receptor Density and Binding Affinity in Healthy Horses and Horses with Systemic Inflammatory Response Syndrome

Background

Dysregulation of the hypothalamic‐pituitary‐adrenal (HPA) axis occurs in horses with systemic inflammatory response syndrome (SIRS). Peripheral resistance to glucocorticoids has not been investigated in horses.

Objective

To determine if glucocorticoid receptor (GR) function in horses can be measured using flow cytometry, and to use this information to evaluate HPA axis dynamics.

Animals

Eleven healthy adult horses in parts 1 and 2. Ten horses with SIRS and 10 age and sex matched controls in part 3.

Methods

Flow cytometry was used to evaluate GR density and binding affinity (BA) in 3 healthy horses in part 1. In part 2, exogenous ACTH was administered to eight healthy horses. Their cortisol response and GR properties were measured. In part 3, CBC, serum biochemistry, cortisol and ACTH, and GR properties were compared between controls without SIRS (n = 10) and horses with SIRS (n = 10), and between survivors and nonsurvivors (n = 4 and n = 6 respectively).

Results

Flow cytometry can be used to measure GR properties in equine PBMCs. No correlation was observed between plasma cortisol concentration and GR density or BA in healthy horses (r = −0.145, P = .428 and r = 0.046, P = .802 respectively). Nonsurvivors with SIRS had significantly decreased GR BA (P = .008). Horses with triglyceride concentration > 28.5 mg/dL had increased odds of nonsurvival (OR=117; 95% CI, 1.94–7,060). GR BA <35.79% was associated with nonsurvival (OR = 30.33; 95% CI, 0.96–960.5).

Conclusions and Clinical Importance

Tissue resistance to glucocorticoids contributes to HPA axis dysfunction in adult horses with SIRS. These horses might benefit from treatment with exogenous glucocorticoids.     

Tumor Necrosis Factor and Interleukin-6 Activity and Endotoxin Concentration in Peritoneal Fluid and Blood of Horses with Acute Abdominal Disease

The purpose of this study was to evaluate the diagnostic and prognostic significance of tumor necrosis factor-alpha (TNF) and interleukin-6 (IL-6) activities and endotoxin concentration in blood and peritoneal fluid of 155 adult horses with acute abdominal disease (colic). Samples also were obtained from 20 healthy adult horses. Blood and peritoneal fluid supernatant TNF and IL-6 activities and endotoxin concentration were significantly greater in horses with colic, compared with healthy horses. In horses with colic, the peritoneal fluid endotoxin concentration and TNF and IL-6 activities were significantly greater than those in blood. Within the colic group, peritoneal fluid IL-6 activity was the analyte that was most frequently increased. Blood and peritoneal fluid supernatant TNF and IL-6 activities were significantly greater when endotoxin was detected in the same sample. Blood and peritoneal fluid IL-6 activity was significantly greater in horses with inflammatory or strangulating lesions, compared with horses having nonstrangulating or noninflammatory lesions. Compared with all other data categories, diagnostic accuracy for nonsurvival was greatest (80%) when blood IL-6 activity exceeded 60 units/mL. The results of this study indicate that endotoxin was present in the peritoneal cavity of at least one third of horses with any acute disease of the abdomen. In horses presented for colic, blood or peritoneal fluid IL-6 activity was more useful than either TNF activity or endotoxin concentration for distinguishing lesion type. Although diagnostic accuracy for the prediction of nonsurvival was good for all of the analytes, negative values were more useful in the prediction of a favorable outcome than were abnormally increased values in the prediction of mortality.     

Haptoglobin Quantitation in Serum Samples from Clinically Normal and Clinically Abnormal Horses

Haptoglobin (HP) is a common minor to moderate acute-phase protein in mammals. It has been described as increasing under a variety of conditions in horses, with the use of different assay methods. The goal of the current study was to provide updated information regarding this biomarker by using a commonly available automated assay. In the present study, reference intervals for HP were generated using 43 serum samples from clinically normal horses. The reference interval was determined via the robust method as 0.29-2.26 mg/ml. A statistically significant 3.3-fold mean increase was observed in HP levels from a clinically abnormal group of horses (n = 33). A weak but significant correlation was found between total white blood cell count and HP levels (r = 0.37, P < .05). A significantly higher level of HP expression was observed in samples acquired from patients whose clinical signs lasted for 7 days or longer than from those whose signs were 4 days or less.     

Plasma and Pulmonary Fluid Endothelin in Horses with Seasonal Recurrent Airway Obstruction

Background: Summer pasture-associated recurrent airway obstruction (SPA-RAO), a seasonal airway obstructive disease of horses, is characterized by clinical exacerbation after exposure to pasture during warm months of the year. Endothelin (ET)-1, potent bronchoconstrictor, mitogen, secretagogue, and proinflammatory mediator, has been implicated in the pathogenesis of asthma and equine heaves.
Hypothesis: Immunoreactive ET-1 concentrations increase during clinical exacerbation and return to basal values during periods of disease remission.
Animals: Twelve horses, 6 affected with SPA-RAO and 6 nonaffected.
Methods: Prospective, observational study. Bronchoalveolar lavage fluid (BALF), arterial and venous plasma samples, and clinical variables were obtained from affected horses during clinical exacerbation and remission. Samples and data of nonaffected horses were collected during the summer and winter on dates similar to affected horses. Immunoreactive ET-1 was determined using a commercial ELISA.
Results: The median and range ET-1 concentrations (pg/ml) in arterial (1.3, 0.7–1.8) and venous (1.3, 1.2–1.7) plasma and in BALF (0.3, 0.2–0.4), and pulmonary epithelial lining fluid (PELF) (25.5, 21–50) were greater in affected horses during clinical exacerbation compared with remission ( P < .01). The concentrations of immunoreactive ET-1 were greater in affected horses during clinical exacerbation compared with nonaffected horses ( P < .05).
Conclusions and Clinical Importance: During clinical exacerbation of SPA-RAO, ET-1 is increased in circulation and pulmonary secretions. Intervention with ET receptor antagonists should provide further information on the role of ET-1 in SPA-RAO.     

Association between Hyperglycemia and Survival in 228 Horses with Acute Gastrointestinal Disease

Background: Glucose homeostasis is dysregulated in critically ill humans resulting in hyperglycemia and decreased survival. Hyperglycemia is common in horses presenting with abdominal crisis, and this might be associated with a worse prognosis for survival.
Objective: To determine if hyperglycemia in horses with acute abdominal disease is associated with increased odds of failure to survive to hospital discharge.
Animals: Two hundred and twenty-eight adult horses with acute gastrointestinal disease.
Methods: Observational retrospective study. Records of horses > 1 year of age presenting for treatment of colic over a 3-year period were reviewed. Data collected included age, duration of colic, glucose, heart rate, PCV, total protein, anion gap, cost of hospitalization, breed, sex, pain at admission, diagnosis, whether surgery was performed, and life status at hospital discharge. Potential risk factors for nonsurvival were screened by univariable logistic regression and the best-fitting univariable model was used as the basis for multivariable regression modeling.
Results: Mean blood glucose was 155 mg/dL (8.5 mM) with 45% of the population above the reference range; 16.7% (38 of 228) of horses had severe hyperglycemia (>195 mg/dL; 10.7 mM). Factors associated with increased odds of failure to survive included glucose, severity of pain at admission, heart rate, PCV, anion gap, and diagnosis. The best-fitting multivariable model included glucose and diagnosis, with age included as a confounding variable. The model correctly classified outcome for 92.5% of horses.
Conclusions and Clinical Importance: This study has confirmed prior reports that hyperglycemia is common in horses with colic and is associated with a worse prognosis for survival to hospital discharge.     

Colic Surgery in Horses: A Retrospective Study Into Short- and Long-Term Survival Rate,Complications and Rehabilitation toward Sporting Activity

Colic surgery in horses impacts both short-term well-being of horses due to possible surgical and anesthetic complications and also long-term return to a sporting career. In this retrospective study, survival and complication rates, as well as functional outcome and behavioral problems in horses that underwent colic surgery were studied. Data from 283 horses that underwent colic surgery at a veterinary teaching hospital were analyzed. Furthermore, owners were contacted and requested to fill out a questionnaire concerning the first year of rehabilitation. Of 283 horses that underwent colic surgery, 167 (59%) were discharged home. After discharge from hospital, 34 horses (12%) were lost to follow-up. Of the remaining 133 horses, 128 were still alive after 1 year (96.2%), while 5 horses were euthanized due to recurrent colic. Of the horses that did not survive the hospitalization period 73 horses (25.8%) were euthanized intraoperatively and 36 horses (12.7%) during intensive care unit (ICU) stay. Survival of horses entering the ICU up to discharge from hospital was 79.5%. During rehabilitation, 49 horses (59.8%) that returned home experienced one or more recurrences of colic. Fifty-two horses (63.4%) that returned home reached at least preoperative level of performance. Altered behavior and gait-related problems during specific elements of riding (for instance during collecting, lateral bending, etc.) were reported in up to 46.2% of horses. Improving veterinary aftercare in collaboration with other disciplines (e.g., physiotherapy and saddle fitting) during rehabilitation could be a means to further improve athletic performance and welfare after recovery from colic surgery.     

Treatment of Age‐Related Hearing Loss in Dogs with the Vibrant Soundbridge Middle Ear Implant: Short‐Term Results in 3 Dogs

Background: Age‐related hearing loss (ARHL), or presbycusis, is the most common form of acquired hearing loss in dogs. Middle ear implants have been used successfully in people with ARHL who cannot benefit from conventional hearing aids. Hypothesis: Audibility improves in dogs with ARHL after implantation of the Vibrant Soundbridge (VSB) middle ear implant. Animals: Three Beagle dogs with ARHL, mean age 11.1 years. Methods: The dogs were assessed pre‐ and postoperatively by brainstem‐evoked response audiometry (BERA), otoscopy, and computed tomography scans of the ears. A VSB middle ear implant was implanted unilaterally. Three months later the functionality of the implants was assessed by auditory steady‐state responses (ASSRs), after which the dogs were euthanized for histopathological examination. Results: The VSB was implanted successfully in all dogs. Recovery from surgery was uneventful, except for transient facial nerve paralysis in 2 dogs. ASSRs showed that hearing improved after activation of the implants with a mean of 20.7, 13, and 16.3 dB at 1, 2, and 4 kHz, respectively. The implantation procedure did not affect residual hearing (with inactive implants) as measured by BERA. Conclusions and Clinical Importance: Implantation of the VSB resulted in lower ASSR thresholds, but only at the higher gain settings of the audioprocessor. As in humans, a more powerful audioprocessor is required to treat sensorineural hearing loss exceeding 20 dB in dogs. A substantial improvement in patient‐owner communication will have to be demonstrated in future studies before the procedure can be recommended in clinical practice.     

Oxidative Stress and Interaction of Endothelin Receptors in Airways of Clinically Healthy Horses

Endothelin-1 (ET-1), a physiological as well as an inflammatory mediator, causes contraction of airway smooth muscles by binding to endothelin-A (ETA) and endothelin-B (ETB) receptors. We investigated the interaction between endothelin receptors on contractions and oxidative stress of bronchial rings of clinically healthy horses. Rings for response studies were set in tissue chambers containing oxygenated Tyrode’s solution. Concentration-response relationships were determined for ET-1 on control rings, rings incubated with ETA antagonist (BQ123), rings incubated with ETB antagonists (IRL 1038 and BQ788), and rings incubated with all antagonists combined. For oxidative stress studies, rings were incubated for 30 minutes separately with ET-1, ETA antagonist + ET-1, ETB antagonists + ET-1, and all three antagonists + ET-1. The control rings were not treated with any agents. Electron paramagnetic resonance (EPR) method was used to measure total reactive oxygen species (ROS), superoxide, and peroxynitrite. Results showed that ET-1 caused dose-dependent contractions which were increased by ETA blockade and decreased by ETB blockade. Combined blockade significantly inhibited the response to ET-1. Regarding the oxidative stress, ET-1 and its antagonists significantly reduced the total ROS. Superoxide production was significantly decreased by ETA antagonist and completely abolished when ETA and ETB antagonists were combined. Peroxynitrite production was not changed significantly. The study suggested that an interaction between ET receptors exists in the expression of contractile responses and oxidative stress. ET-1 attenuated oxidative stress by decreasing total ROS. ETA receptors may be primarily responsible for superoxide production. It appears that ET-1 does not affect peroxynitrite production in normal equine tissues.