Measuring V̇O2 in hypoxic and hyperoxic conditions using dynamic gas mixing with a flow-through indirect calorimeter.

Measurements of gas exchange while breathing gases of different O2 concentrations are useful in respiratory and exercise physiology. High bias flows required in flow-through indirect calorimetry systems for large animals like exercising horses necessitate the use of inconveniently large reservoirs of mixed gases for making such measurements and can limit the amount of equilibration time that is adequate for steady-state measurements. We obviated the need to use a pre-mixed reservoir of gas in a semi-open flow-through indirect calorimeter by dynamically mixing gases and verified the theoretical accuracy and utility of making such measurements using the mass-balance N2-dilution method. We evaluated the accuracy of the technique at different inspired oxygen fractions by measuring exercising oxygen consumption (V̇O2) at two fully aerobic submaximal exercise intensities in Thoroughbred horses. Horses exercised at 24% and 50% maximum oxygen consumption (V̇O2 max) of each horse while breathing different O2 concentrations (19.5%, 21% and 25% O2). The N2-dilution technique was used to calculate V̇O2. Repeated-measures ANOVA was used to tested for differences in V̇O2 between different inspired O2 concentrations. The specific V̇O2 of the horses trotting at 24%V̇O2max and cantering at 50%V̇O2max were not significantly different among the three different inspired oxygen fractions. These findings demonstrate that reliable measurements of V̇O2 can be obtained at various inspired oxygen fractions using dynamic gas mixing and the N2-dilution technique to calibrate semi-open-circuit gas flow systems.

The Effects of Equine Assisted Therapy on Plasma Cortisol and Oxytocin Concentrations and Heart Rate Variability in Horses and Measures of Symptoms of Post-Traumatic Stress Disorder in Veterans.

With the increase in the number of horses being used in Equine-Assisted Activities and Therapies (EAAT) programs and with the increasing concern for animal welfare, it is important to understand the impact of such interventions on the stress level and quality of life for the horses involved. The purpose of the present pilot study was to test the hypothesis that participation in EAAT would acutely alter physiological markers of stress and well-being, including plasma cortisol, plasma oxytocin, and heart rate variability (HRV), in horses and that symptoms of posttraumatic stress disorder (PTSD) would be reduced after five sessions of EAAT in veterans who had previously been diagnosed with PTSD. Nine healthy geldings, of various breeds, ages 10-23 years, conditioned and experienced as therapeutic riding horses, were selected to participate in the study. Of these, seven were selected at random to wear electrocardiogram units, and all nine were used for blood sampling to measure plasma cortisol and oxytocin. Each horse was randomly assigned to partner with a veteran for five EAAT sessions, 1 hour in duration. A standing control was conducted on a later date on which horses did not participate in EAAT. Measurement after 5 days of EAAT was conducted immediately after the end of the last session on day 5 using the Brief Symptom Inventory and the PCL-5 (a 20 item self-report measure of the Diagnostic and Statistical Manual of Mental Disorders - 5 for symptoms of PTSD). Two way repeated measure analysis of variance showed no significant day by time interactions for plasma cortisol (P = .821) or oxytocin (P = .861). There was a significant day by time interaction (P = .006) for heart rate (HR); where on day 1, HR (bpm) was significantly lower during the interaction with the veterans. There were no significant differences in HRV variables. Posttherapy measures in PTSD symptoms in veterans were significantly reduced except for interpersonal sensitivity (P = .08) and phobic anxiety (P = .17). There was an effect of EAAT on HR which was significantly reduced on day 2 during the actual EAAT session. Equine-Assisted Activities and Therapies had no effect on respiration rate and systolic or diastolic blood pressure in veterans involved in five sessions of EAAT, lasting 60 minutes in duration over the course of 5 days. Stress levels, as demonstrated by plasma cortisol concentrations and HRV, did not change in horses involved in EAAT sessions with veterans who had been previously diagnosed with PTSD. Furthermore, the horses used in this study did not demonstrate increased levels of well-being as demonstrated by the lack of change in plasma oxytocin concentrations after EAAT sessions. Symptoms of PTSD did change significantly in the veterans who participated in this study.

Comparison of echocardiographic measurements in elite and nonelite Arabian endurance horses.

OBJECTIVE: To determine whether echocardiographic variables differed between successful (elite) and less successful (nonelite) Arabian endurance horses. ANIMALS: 34 Arabian horses that competed in endurance racing. PROCEDURES: Horses were assigned to either an elite or nonelite group on the basis of results of a previous competition, and a standardized echocardiographic examination was performed on each horse within 1 to 4 weeks after that competition. Multivariable logistic regression with backward stepwise elimination was used to create a prediction model for the determination of horse status (elite or nonelite) as a function of the measured echocardiographic variables. RESULTS: The elite and nonelite groups consisted of 23 and 11 horses, respectively. One horse in the nonelite group had a frequent ventricular dysrhythmia that could have negatively affected its performance and rider's safety, whereas none of the horses in the elite group had remarkable cardiac abnormalities. The left ventricular internal diameter during systole and diastole and left ventricular mass and stroke volume were significantly greater for horses in the elite group, compared with those for horses in the nonelite group. The final logistic regression model correctly predicted the horse status for all of the horses in the elite group and 8 of 11 horses in the nonelite group. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that heart size was significantly associated with performance for Arabian endurance horses in a manner similar to findings for Thoroughbred and Standardbred racehorses in active competition.

Cardiac troponin I concentrations in horses with colic.

OBJECTIVE: To determine prevalence of myocardial injury in horses with colic on the basis of high concentrations of cardiac troponin I (cTnI), frequency of cardiac arrhythmias within the first 24 to 48 hours after hospital admission or surgery because of colic, and associations between high cTnI concentrations and cardiac arrhythmias, clinical course, and outcome (survival to discharge from hospital vs nonsurvival [death or euthanasia]). DESIGN: Prospective observational study. ANIMALS: 111 horses with colic. PROCEDURES: Blood was drawn at admission and 12 and 24 hours after admission if horses were treated medically or 12 and 24 hours after surgery if treated surgically. A 24-hour ambulatory ECG was recorded beginning the morning after admission in medically treated cases or after surgery and evaluated for arrhythmias. Clinical and clinicopathologic data and outcome were obtained. Associations between cTnI concentrations and other variables were determined. RESULTS: An abnormal cTnI concentration (≥ 0.10 ng/mL) at admission was significantly associated with the occurrence of ventricular arrhythmias, outcome, and surgical treatment. CONCLUSIONS AND CLINICAL RELEVANCE: The data suggested that horses with colic and high cTnI concentrations at admission were more likely to have ventricular arrhythmias and have a less favorable prognosis for recovery. High cTnI concentrations in horses with colic were suggestive of myocardial damage.

A modified laryngoplasty approach promoting ankylosis of the cricoarytenoid joint.

OBJECTIVE: To perform a modification to the standard laryngoplasty procedure in vivo that would result in ankylosis of the cricoarytenoid (CA) joint, and determine the stability provided to the abducted arytenoid in vitro. STUDY DESIGN: Experimental study. ANIMALS: Horses (n=8). METHODS: Horses were assigned to either control laryngoplasty (n=3) or modified laryngoplasty (5) procedure. Endoscopic upper airway evaluations were used to measure right:left quotients 1 day and 3 months postoperatively to assess maintenance of abduction. Horses were euthanatized 3 months after surgery and larynges collected for measurement of translaryngeal impedance and histologic evaluation of CA joint ankylosis. Each specimen was exposed to increasing negative pressure with the sutures intact or cut while translaryngeal impedance was recorded. Data were analyzed using ANOVA with significance set at P<.05. RESULTS: Loss of left arytenoid cartilage abduction at 3 months was greater in the control laryngoplasty group. Overall, impedance was significantly lower for the modified laryngoplasty group compared with the control laryngoplasty group and lower with the sutures intact than cut. Histologic evaluation of the joints confirmed fibrous bridging of the left CA joints of the modified laryngoplasty group. CONCLUSIONS: A modified laryngoplasty approach promotes ankylosis of the CA joint and decreases the loss of abduction of the arytenoid.

Cardiac output determination by use of lithium dilution during exercise in horses.

OBJECTIVE: To compare cardiac output (CO) obtained by the lithium dilution method (LiDCO) with CO calculated from the Fick principle (FickCO), in horses maximally exercising on a high-speed treadmill. ANIMALS: 13 Thoroughbreds. PROCEDURES: In part 1 of the study, 5 horses performed a warm-up (walk, trot, and canter) and exercise test (walk, trot, canter, and gallop [90% to 100% maximum oxygen consumption [{VO(2)max}]) with measurements of LiDCO and FickCO obtained simultaneously after 60 seconds at each exercise level, for a total of 7 measurements. In part 2 of the study, 8 horses performed a warm-up (walk, trot, and canter) followed by an exercise test (walk and gallop [90% to 100% [VO(2)max], repeated twice). Measurements of LiDCO and FickCO were obtained 60 seconds into the first walk and each gallop of the exercise tests, for a total of 3 measurements. RESULTS: Cardiac output increased significantly with increasing speeds by use of both methods. In part 1, lithium dilution significantly overestimated CO, compared with the Fick principle, during the exercise test (as both injection number and exercise intensity increased). Mean +/- SD bias was 246 +/- 264 mL of blood/min/kg in part 1 and 67 +/- 100mL of blood/kg/min in part 2. Three injections of lithium (part 2) did not result in the same degree of overestimation of LiDCO that was observed with 7 injections (part 1). CONCLUSIONS AND CLINICAL RELEVANCE: Lithium dilution may be an acceptable substitute for the Fick principle as a means to measure CO in maximally exercising client-owned horses.

Cardiac output technologies with special reference to the horse.

Critical illness, anesthesia, primary cardiovascular disease, and exercise may result in marked hemodynamic alterations. Measuring cardiac output (CO) is central to defining these alterations for both clinician and researcher. In the past 10 years, several new methods of measuring CO have been developed for the human medical market. Some of these methods are now validated in the horse and are in clinical use. The Fick method has been used in equine research for more than a century. It depends on simultaneous measurement of mixed venous (pulmonary arterial) and peripheral arterial oxygen content and oxygen uptake by the lungs. The technique is technically demanding, which restricts its clinical use. Indicator dilution techniques, with indocyanine green, cold (thermodilution), or lithium as the marker, have also been widely used in the horse. The indocyanine technique is cumbersome, and thermodilution requires right heart catheterization, which is not a benign procedure, making both of these methods less than ideal for clinical use. Lithium dilution requires catheterization of a peripheral artery and a jugular vein. It has recently been validated in anesthetized adult horses and neonatal foals. Doppler echocardiography is a noninvasive ultrasound-based technique. More accurate measurements are obtained with transesophageal than with transthoracic measurements; however, both methods require considerable technical expertise. Bioimpedance and pulse contour analysis are 2 new methods that have yet to be validated in the horse. With the currently available technology, lithium dilution appears to be the method of measuring CO best suited to the equine clinic.

Exercise-induced pulmonary hemorrhage.

EIPH is a condition affecting virtually all horses during intense exercise worldwide. The hemorrhage originates from the pulmonary vasculature and is distributed predominantly bilaterally in the dorsocaudal lung lobes. As the condition progresses, the lung abnormalities extend cranially along the dorsal portions of the lung. An inflammatory response occurs in association with the hemorrhage and may contribute to the chronic sequela. Although conflicting opinions exist as to its affect on performance, it is a syndrome that is thought to increase in severity with age. The most commonly performed method to diagnose EIPH at the present time is endoscopy of the upper airway alone or in combination with tracheal wash analysis for the presence of erythrocytes and hemosiderophages. Because horses may not bleed to the same extent every time and the bleeding may originate from slightly different locations, these diagnostic procedures may not be extremely sensitive or quantitative. At this time, there is no treatment that is considered a panacea, and the currently allowed treatments have not proven to be effective in preventing EIPH. Future directions for therapeutic intervention may need to include limiting inflammatory responses to blood remaining within the lungs after EIPH.

Quantification of clenbuterol in equine plasma, urine and tissue by liquid chromatography coupled on-line with quadrupole time-of-flight mass spectrometry.

Clenbuterol (CBL) is a potent beta(2)-adrenoceptor agonist used for the management of respiratory disorders in the horse. The detection and quantification of CBL can pose a problem due to its potency, the relatively low dose administered to the horse, its slow clearance and low plasma concentrations. Thus, a sensitive method for the quantification and confirmation of CBL in racehorses is required to study its distribution and elimination. A sensitive and fast method was developed for quantification and confirmation of the presence of CBL in equine plasma, urine and tissue samples. The method involved liquid-liquid extraction (LLE), separation by liquid chromatography (LC) on a short cyano column, and pseudo multiple reaction monitoring (pseudo-MRM) by electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-QTOF-MS/MS). At very low concentrations (picograms of CBL/mL), LLE produced better extraction efficiency and calibration curves than solid-phase extraction (SPE). The operating parameters for electrospray QTOF and yield of the product ion in MRM were optimized to enhance sensitivity for the detection and quantification of CBL. The quantification range of the method was 0.013-10 ng of CBL/mL plasma, 0.05-20 ng/0.1 mL of urine, and 0.025-10 ng/g tissue. The detection limit of the method was 13 pg/mL of plasma, 50 pg/0.1 mL of urine, and 25 pg/g of tissue. The method was successfully applied to the analysis of CBL in plasma, urine and various tissue samples, and in pharmacokinetic (PK) studies of CBL in the horse. CBL was quantified for 96 h in plasma and 288 h in urine post-administration of CLB (1.6 micro g/kg, 2 x daily x 7 days). This method is useful for the detection and quantification of very low concentrations of CBL in urine, plasma and tissue samples.