Atrial, SA nodal, and AV nodal electrophysiology in standing horses: normal findings and electrophysiologic effects of quinidine and diltiazem

BACKGROUND: Although atrial arrhythmias are clinically important in horses, atrial electrophysiology has been incompletely studied. HYPOTHESES: Standard electrophysiologic methods can be used to study drug effects in horses. Specifically, the effects of diltiazem on atrioventricular (AV) nodal conduction are rate-dependent and allow control of ventricular response rate during rapid atrial pacing in horses undergoing quinidine treatment. ANIMALS: Fourteen healthy horses. METHODS: Arterial blood pressure, surface electrocardiogram, and right atrial electrogram were recorded during sinus rhythm and during programmed electrical stimulation at baseline, after administration of quinidine gluconate (10 mg/kg IV over 30 minutes, n = 7; and 12 mg/kg IV over 5 minutes followed by 5 mg/kg/h constant rate infusion for the remaining duration of the study, n = 7), and after coadministration of diltiazem (0.125 mg/kg IV over 2 minutes repeated every 12 minutes to effect). RESULTS: Quinidine significantly prolonged the atrial effective refractory period, shortened the functional refractory period (FRP) of the AV node, and increased the ventricular response rate during atrial pacing. Diltiazem increased the FRP, controlled ventricular rate in a rate-dependent manner, caused dose-dependent suppression of the sinoatrial node and produced a significant, but well tolerated decrease in blood pressure. Effective doses of diltiazem ranged from 0.125 to 1.125 mg/kg. CONCLUSIONS AND CLINICAL IMPORTANCE: Standard electrophysiologic techniques allow characterization of drug effects in standing horses. Diltiazem is effective for ventricular rate control in this pacing model of supraventricular tachycardia. The use of diltiazem for rate control in horses with atrial fibrillation merits further investigation.     

Effect of Transvenous Electrical Cardioversion on Plasma Cardiac Troponin I Concentrations in Horses with Atrial Fibrillation

Background: Whether electrical cardioversion of cardiac arrhythmias results in cardiomyocyte damage is unknown.
Objective: To describe effect of transvenous electrical cardioversion (TVEC) on plasma cardiac troponin I (cTnI) concentration in horses.
Animals: All horses presented to the Cornell University Hospital for Animals for cardioversion of atrial fibrillation between May 2006 and October 2008 were eligible for inclusion in the study. Owners of 14 horses elected for TVEC and each horse was then enrolled (16 procedures).
Methods: Prospective observational study measuring concentrations of plasma cTnI before and after TVEC.
Results: Median cTnI concentration increased from 0.045 ng/mL at baseline (range 0.0–0.20 ng/mL) to 0.11 ng/mL after TVEC (range 0.0–3.73 ng/mL) ( P = .036). This increase was not associated with the number of shocks delivered, maximal energy delivered, cumulative energy delivered, chronicity of atrial fibrillation before cardioversion, or positioning of the pulmonary artery catheter.
Conclusions: The increase in cTnI is unlikely to be clinically important. The increase might be correlated with persistent atrial dysfunction after TVEC, suggesting that a longer convalescent period after the procedure could be warranted.     

Plasma Procalcitonin Concentration in Healthy Horses and Horses Affected by Systemic Inflammatory Response Syndrome

Background

The diseases most frequent associated with SIRS in adult horses are those involving the gastrointestinal tract. An early diagnosis should be the goal in the management of horses with SIRS.

Objective

The objective of this study was to evaluate the plasma procalcitonin (PCT) concentration in healthy and SIRS horses to assess differences between the two groups.

Animals

Seventy‐eight horses (30 healthy and 48 SIRS).

Methods

Prospective in vivo multicentric study. Horses were classified as SIRS if at least 2 of the following criteria were met: abnormal leukocyte count or distribution, hyperthermia or hypothermia, tachycardia, tachypnea. Healthy horses showed no clinical or laboratory signs of SIRS. Plasma PCT concentrations were measured with a commercial ELISA assay for equine species. Results were expressed as mean±standard deviation. T‐test for unpaired data was performed between healthy and SIRS group. SIRS group was divided in 4 subgroups and t‐test was performed between healthy versus each subgroup.

Results

PCT concentrations in healthy and SIRS horses were 18.28 ± 20.32 and 197.0 ± 117.0 pg/mL, respectively. T‐test showed statistical differences between healthy versus SIRS group and between healthy versus all subgroups.

Conclusions and Clinical Importance

Results showed an increase in PCT concentration in SIRS horses as previously reported in humans and dogs. PCT could be used as a single assay in equine practice for detection of SIRS.     

Cerebral and Brainstem Electrophysiologic Activity During Euthanasia with Pentobarbital Sodium in Horses

Background

An overdose of pentobarbital sodium administered IV is the most commonly used method of euthanasia in veterinary medicine. Determining death after the infusion relies on the observation of physical variables. However, it is unknown when cortical electrical activity and brainstem function are lost in a sequence of events before death.

Hypothesis/Objectives

To examine changes in the electrical activity of the cerebral cortex and brainstem during an overdose of pentobarbital sodium solution for euthanasia. Our testing hypothesis is that isoelectric pattern of the brain in support of brain death occurs before absence of electrocardiogram (ECG) activity.

Animals

Fifteen horses requiring euthanasia.

Methods

Prospective observational study. Horses with neurologic, orthopedic, and cardiac illnesses were selected and instrumented for recording of electroencephalogram, electrooculogram, brainstem auditory evoked response (BAER), and ECG. Physical and neurologic (brainstem reflexes) variables were monitored.

Results

Loss of cortical electrical activity occurred during or within 52 seconds after the infusion of euthanasia solution. Cessation of brainstem function as evidenced by a lack of brainstem reflexes and disappearance of the BAER happened subsequently. Despite undetectable heart sounds, palpable arterial pulse, and mean arterial pressure, recordable ECG was the last variable to be lost after the infusion (5.5–16 minutes after end of the infusion).

Conclusions and Clinical Importance

Overdose of pentobarbital sodium solution administered IV is an effective, fast, and humane method of euthanasia. Brain death occurs within 73–261 seconds of the infusion. Although absence of ECG activity takes longer to occur, brain death has already occurred.     

Clinical Feasibility and Airway Deposition of Nebulized Voriconazole in Healthy Horses

Voriconazole (VRC) is a potential treatment for pneumomycosis in horses. The objectives of this study were to determine if the delivery of Vfend using a Flexineb nebulizer produced clinically significant [VRC] in lower airways. The hypothesis was that [VRC] after delivery by nebulization would be greater in the pulmonary epithelial lining fluid than plasma. A secondary objective was to determine [VRC] in upper airways through the collection of nasopharyngeal wash (NPW) samples. Voriconazole solution [Vfend-6.25 mg/mL, 100 (n = 2), 200 (n = 3), 500 (n = 1) mg] was nebulized once in 6 healthy geldings. Clinical responses, duration of nebulization, and [VRC] at various time points (up to 8 hours) in plasma, bronchoalveolar lavage fluid (BALF) supernatant and cell pellet, and NPW samples were recorded. Voriconazole (Vfend-6.25 mg/mL, 200 mg) was nebulized in 5 additional, healthy geldings, and [VRC] was measured in NPW samples pre- and postnebulization at time points up to 8 hours. The antifungal activity of BALF and NPW samples was determined using agar disk diffusion. Concentrations of voriconazole were below detection in plasma, BALF supernatant, and cell pellets for all time points and doses except the BALF cell pellet (0.4 μg/g) immediately after nebulization of 500 mg. For 5 horses, administered 200 mg of Vfend, mean [VCR] in NPW at the end of nebulization and 1, 6, and 8 hours postnebulization were: 30.8 ± 29, 1.0 ± 0.84, 0.2 ± 0.19, and 0.34 ± 0.67 μg/mL, respectively. Only NPW samples obtained immediately postnebulization showed antifungal activity. A nebulized Vfend solution is not recommended for the treatment of pneumomycosis in horses.     

Electrophysiologic Characteristics and Topographic Distribution of Focal Atrial Tachycardias in Dogs

Background: Focal atrial tachycardia (FAT) is a common supraventricular tachycardia in dogs. Objective: To evaluate electrophysiologic characteristics and topographic distribution of FAT. Animals: Sixteen dogs with symptomatic FAT. Methods: Retrospective case series. Electrophysiological studies were performed to test the inducibility of documented and no documented arrhythmias. Once induced for each dog, FAT was analyzed for electrogenic mechanism, endocardial electrogram, and location. Results: Nineteen FATs could be studied in 16 dogs, 12 were automatic, 4 nonautomatic, and 3 incessant. Two dogs had >1 focus. Mean atrial cycle length (CL) was 238.2 ± 69.2 (SD) milliseconds, mean ventricular CL of 292.7 ± 72.5 (SD) milliseconds, with atrioventricular block in 6 cases. Mean presystolic atrial activity recorded at the ectopic focus was –39.9 ± 17.7 (SD) milliseconds. Atrial potentials were fragmented in 11 dogs and were low amplitude in 6 dogs. Sixty‐three percent of ectopic foci were distributed within the right atrium (5 crista terminalis, 3 triangle of Koch, 2 tricuspid valve annulus, 1 interatrial septum, and 1 right auricle) and 37% in the pulmonary veins (PVs) (4 right superior PV, 2 left superior PV, and 1 right inferior PV). Persistent atrial fibrillation (AF) and paroxysmal AF were triggered by FATs in 7 dogs (2 with multiple ectopic foci and 4 with at least one PV focus). Conclusion and Clinical Relevance: According to our findings, dogs have a predominance of right‐sided FAT. The majority of FATs are automatic and can trigger AF, particularly in the case of PV location.     

Comparison of Insulin-Induced Digital Vessel Ring Responses of Laminitic and Clinically Healthy Horses

Hyperinsulinemia leads to insulin resistance of blood vessels and interferes with circulation of laminae of the equine foot. The objective of the study was to compare the digital vessel responses of clinically healthy and laminitic horses with and without experimental induction of insulin resistance. Vessel segments (artery and vein) collected after euthanasia were cut into 3-mm wide rings and were prepared in tissue baths containing Tyrode solution for response studies. Two rings of each vessel type from each horse were used as nonresistant, and two were made insulin resistant. Insulin resistance was induced by incubating the rings with 10-μM bath concentration of insulin. Then, all rings were contracted with phenylephrine (5 μM). When the contraction reached a plateau, insulin (10 μM) was added, and the response monitored for 30 minutes. The response to insulin was calculated as a percentage of the phenylephrine response. The results showed that the responses of the nonresistant rings differed significantly between the groups. However, when all rings were made insulin resistant, the significance between the groups disappeared. In the laminitic group, responses of resistant and nonresistant rings did not differ, whereas in the healthy group, the vessel responses differed significantly. In all experiments, arterial and venous rings followed the same pattern, but the magnitudes were greater in arterial rings. The findings suggest that the vessel responses to insulin are altered in laminitis which could be due to insulin resistance.     

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.     

Effects of acute gastric distention and recovery on tendency for ventricular arrhythmia in dogs

The gastric distention-volvulus (GDV) syndrome occurs commonly in large-breed dogs and may prove fatal in 15-68% of cases. Approximately 43% of cases with gastric distention (GD) or volvulus develop cardiac arrhythmias that can contribute to mortality. Most of these arrhythmias are ventricular in origin and ventricular fibrillation (VF) may be the cause of death. This study used an iatrogenic model of acute GD to investigate the prevalence of ventricular arrhythmias during acute GD and its recovery, if programmed electrical stimulation (PES) may uncover tendency to VF, if the Q-T interval corrected for heart rate (Q-Tc) of the electrocardiogram (ECG) predicts tendency to VF, and if hemodynamic changes predate VF. Eleven beagles, anesthetized with morphine and alpha-chloralose, and instrumented so that vascular pressures, cardiac output, and PES could be recorded, were used. Five were unperturbed, whereas acute GD to a pressure of 30 mm Hg for 1.5 hours was produced in 6 others. The results were as follows. No dogs with GD developed spontaneously occurring arrhythmias. VF was produced in no dogs by conventional PES, but occurred in all dogs (P < .05) with GD and none of the controls, using accelerated ventricular pacing. The Q-Tc interval of the ECG prolonged minimally in dogs with GD, and shortened (P < .05) in controls. Some hemodynamic changes did predate VF. In conclusion, dogs with acute GD have a tendency for VF, which may be uncovered by accelerated PES. The mechanism for the vulnerability to arrhythmia with GD is unknown.     

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.     

Atrial and Ventricular Electrical and Contractile Remodeling and Reverse Remodeling Owing to Short‐Term Pacing‐Induced Atrial Fibrillation in Horses

Background: In humans, atrial fibrillation (AF) induces electrical, contractile, and structural remodeling leading to AF stabilization. Little is known about AF‐induced atrial remodeling in horses. Hypothesis: Induced AF produces rapid atrial electrical and contractile remodeling in horses. Animals: Six horses, 5 animals completed the study. Methods: Each horse was instrumented with a pulse generator and pacemaker to maintain AF by burst pacing and to study atrial and ventricular electrophysiology (AF cycle length [AFCL], AF duration, and atrial/ventricular effective refractory period [AERP/VERP] at different pacing cycle lengths [PCL]). Left atrial and ventricular contractile remodeling were assessed echocardiographically by calculation of fractional changes in atrial and ventricular dimensions, respectively, during the cardiac cycle. Measurements were performed at baseline, a 7‐day AF period and a 2‐day recovery period. Results: Atrial electrical and contractile remodeling could be demonstrated after 4 and 12 hours of AF, respectively. A progressive shortening of the AERP (261 ± 39–171 ± 18 ms at a PCL of 1,000 ms, P < .0001), an attenuation of the AERP rate adaptation, a decrease in AFCL (239 ± 39–194 ± 7 ms, P < .0001), and a decrease in atrial FS (12 ± 3% to 0 ± 2%, P < .05) occurred. AF duration increased progressively and became persistent in 2 animals. VERP did not change significantly. Upon restoration of sinus rhythm, values returned to baseline within 48 hours. Conclusions and Clinical Importance: Atrial electrical and contractile remodeling appears rapidly. After 7 days of AF, reverse remodeling occurred within 2 days. These observations suggest that early conversion of AF might be beneficial for success rate and early return to training.     

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.     

Effects of Blood Contamination on Peritoneal D-Dimer Concentration in Horses with Colic

Background: Peritoneal D-Dimer concentration can be determined to assess peritoneal fibrinolysis activity in horses with gastrointestinal disorders. However, blood contamination of peritoneal fluid may occur during collection and could alter peritoneal D-Dimer concentration.
Hypothesis/Objectives: Blood contamination in peritoneal fluid does not affect interpretation of peritoneal D-Dimer concentration in horses with colic.
Animals: Thirty-four horses with colic and 4 healthy horses.
Methods: Peritoneal fluid and blood samples were simultaneously collected upon admission. Then, peritoneal fluid was serially contaminated with the horse's own blood; final contaminations corresponded to 1, 5, 10, and 20% of blood in peritoneal fluid. D-Dimer concentration was determined in blood, peritoneal fluid, and contaminated peritoneal fluid samples. Data were analyzed using a longitudinal linear model and a generalized estimating equations analysis to assess the quantitative and qualitative variations of the effect of blood contamination on peritoneal D-Dimer concentration.
Results: Peritoneal D-Dimer concentration was only quantitatively affected when peritoneal fluid was contaminated at 20% of blood. However, when using increasing cut-off values of peritoneal D-Dimer concentration (100, 2,000, 8,000, and 16,000 ng/mL), this effect disappeared at the highest cut-off values (8,000 and 16,000 ng/mL). When peritoneal fluid contamination was grouped as "minimally contaminated" (≤1% of blood) and "highly contaminated" (≥5% of blood), no significant differences on D-Dimer concentration between both groups at each cut-off value were observed.
Conclusions and Clinical Importance: Although quantitative results of peritoneal D-Dimer concentration could be affected by high levels of blood contamination (≥20%), interpretation of increased peritoneal fibrinolytic activity was not significantly affected.     

The Use of Ultrasound to Assess the Thrombogenic Properties of Teflon and Polyurethane Catheters for Short-Term Use in Systemically Healthy Horses

The objective of this study was to establish an ultrasound scoring system to assess the degree of fibrin tag and clot formation associated with jugular catheters in horses. A second objective was to test the hypothesis that, in healthy horses undergoing short-term anesthesia, polyurethane (PU) catheters have a different ultrasound score compared with polytetrafluoroethylene (Teflon) catheters. Twenty-three systemically healthy horses requiring anesthesia were randomly assigned PU or Teflon catheters. The mean catheter dwell time was 5 hours. Ultrasound was used to examine the jugular vein before catheter placement, before catheter removal (mean, 4 hours 45 minutes), and approximately 24 hours after catheter insertion (mean, 24 hours 42 minutes). The repeatability of the ultrasound scoring system was assessed by producing a series of randomly ordered selected still images for independent evaluation by four radiologists on two separate occasions. The interobserver and intraobserver repeatability was determined using a weighted kappa statistic, with substantial intraobserver repeatability mean κ .77 (range, .59–.91) and moderate interobserver repeatability mean κ .53 (range, .33–.67). The proportion of positive bacteriologic cultures in the polyurethane (PU) and Teflon groups was compared using Fisher's exact test, with no significant difference (P > .99) between the two groups. The distribution of the scores from the PU and Teflon groups was compared using the Mann-Whitney test, with no evidence of a difference between groups at the time of catheter removal (P = .23) and 24 hours after catheter insertion (P = .20). Further studies with a larger sample of systemically healthy horses are required to substantiate this preliminary finding.     

Results of Quantitative Cultures of Urine by Free Catch and Catheterization from Healthy Adult Horses

Quantitative urine cultures were performed on 11 male and 11 female healthy adult horses. Urine was collected by free catch and catheterization using standard methods. Results showed that all samples collected by free catch contained less than 20,000 colony-forming units (CFU)/mL. All samples collected by catheterization contained 500 CFU/mL or less. A significant difference was found between collection methods ( P < .005), with catheterization having less contamination. In samples collected by free catch, females had significantly greater contamination than did males ( P < .03). Predominant bacterial species isolated included Streptococcus spp., Escherichia coli, Enterohacter sp., Bacillus sp., Staphylococcus spp., Diptheroids sp., Proteus spp., and Enterococcus sp. Many samples contained multiple bacterial species. Bacterial isolates were representative of the normal bacterial flora of the equine urogenital tract. This paper establishes reference values for quantitative urine culture results in healthy adult horses to aid in the diagnosis of urinary tract infections.