Preliminary report into the function of the shoulder using a novel imaging and motion capture approach.

REASONS FOR PERFORMING STUDY: The function of the forelimb is fundamental to understanding both sound and pathological locomotion. The precise movements of the equine shoulder are hidden by layers of skin and muscle and hence the shoulder is normally modelled as a simple pivot during locomotion which assumes that any translational motion is negligible. OBJECTIVES: To record and quantify the sliding motion of the scapula during locomotion, using a novel imaging technique. METHODS: Scapula motion during locomotion in the horse was calculated by tracking the ripple of the shoulder blade's movement under an array of markers placed over the soft tissue. RESULTS: Interstride variability was low. Sliding of up to 80 mm in the plane of progression (cranio-caudal) was observed; however, the limits of motion varied by <5 mm in the gaits examined, despite variations in stride length. Stride length appeared to be increased by scapula rotation in the plane of progression, and this flexion-extension was largest in trot and was not significantly different between walk and canter. This was in agreement with the distance travelled by the trunk whilst the hoof was on the ground. Substantial sliding in a dorsal-ventral direction was shown and varied with the gait used, both in magnitude and timing, possibly providing a shock absorption mechanism. The sliding did not increase as much as would be expected in canter and this coincided with a more lateral positioning of the scapula and increased impact on the ribcage. CONCLUSIONS: It has been assumed that scapula-thoracic sliding increases stride length and hence economically increases locomotor speed. The extra motion of the scapula recorded appeared to absorb shock from forelimb impact and maintain the economy of locomotion, but did not increase with speed and the muscular pretensioning implied could actually impair ventilation.

Efficacy of thermal cautery for intermittent dorsal displacement of the soft palate as compared to conservatively treated horses: results from 78 treadmill diagnosed horses.

REASONS FOR PERFORMING STUDY: Previously, objective comparisons of surgical procedures to relieve dorsal displacement of the soft palate (DDSP) have been limited by the presumptive basis of the diagnostic measures applied. OBJECTIVES: To assess and compare the efficacy of thermal cautery surgery to conservatively treated controls in racehorses definitively diagnosed with idiopathic intermittent DDSP. HYPOTHESIS: Both conservative and surgical treatments have a beneficial result on racing performance in racehorses affected with DDSP. METHODS: Race records were obtained for Thoroughbred racehorses definitively diagnosed with DDSP using high-speed treadmill endoscopy. Racing performance was assessed based on prize money won. RESULTS: Forty-eight horses that underwent thermal cautery and 30 conservatively treated controls were included. Pretreatment earnings significantly decreased in the race immediately prior to diagnosis. A high proportion of previously raced horses returned to racing after both treatments (90-96%). Intrahorse comparison of earnings in 3 races pre- vs. post treatment showed that 53% of conservatively treated horses and 36% of the thermal cautery group had improved performance. Although the difference between these 2 groups may be interpreted as being clinically significant, it was not statistically significant. CONCLUSIONS AND POTENTIAL RELEVANCE: A higher percentage of conservatively treated controls had improved individual performance compared to horses treated with thermal cautery. Thermal cautery appears less effective than other previously published surgical treatments for DDSP. Comparison of the 2 treatment methods should be interpreted cautiously because treatments were not randomised, resulting in baseline variability between groups.

Organic dust exposure increases mast cell tryptase in bronchoalveolar lavage fluid and airway epithelium of heaves horses.

BACKGROUND: Mast cell degranulation is believed to act as a key event in initiating and maintaining airway response to allergen challenge in human asthma. It is hypothesized that the mast cell may play a similar role in equine heaves, which shares many similarities with occupational dust-induced asthma. OBJECTIVE: The aim of this study was to quantify the mast cell proteinase tryptase in bronchoalveolar lavage fluid (BALF) from control and heaves-susceptible horses and to investigate tryptase mRNA and protein expression in pulmonary mast cells. METHODS: Equine BALF tryptase concentrations were determined by ELISA from control and heaves-susceptible horses pre and post 24 h hay/straw challenge (HSC). Tryptase mRNA and protein expression were investigated by quantitative PCR and immunohistochemistry in bronchial and bronchiolar tissue samples of control and heaves-susceptible horses. RESULTS: Both control and heaves-susceptible horses had significantly increased BALF tryptase concentrations following HSC (P=0.003 and 0.034, respectively). Increased numbers of tryptase-expressing intra-epithelial mast cells were demonstrated in heaves horses, but not controls, following challenge (P=0.02). Bronchiolar tissue from heaves horses removed from challenge contained significantly lower tryptase transcripts than that from control horses (P=0.02). CONCLUSION: Mast cell degranulation and tryptase release into the airways occur following HSC of control and heaves-susceptible horses. The greater number of mast cells available in the bronchiolar epithelium of heaves horses may be clinically significant in the pulmonary inflammatory response of heaves.

Comparison of the antioxidant status in tracheal and bronchoalveolar epithelial lining fluids in recurrent airway obstruction.

REASONS FOR PERFORMING STUDY: Following a period of airway inflammation the clearance of inflammatory cells along the mucociliary escalator may impose a considerable oxidant load on the trachea. OBJECTIVES: To determine the degree of oxidative stress in tracheal epithelial lining fluid (ELF) in comparison to that present in peripheral airways after an acute exposure to organic dust. METHODS: Tracheal wash fluid and bronchoalveolar lavage fluid (BALF) were collected for cytology and antioxidant analyses from 6 recurrent airway obstruction (RAO)-affected horses and 6 healthy control horses before and after stabling on straw bedding for 24 h. RESULTS: In RAO-affected horses, organic dust exposure resulted in a significant decrease in ascorbic acid concentration in tracheal ELF (P<0.0001), which was greater than the decrease in bronchoalveolar ELF (P = 0.0003). The percentage decrease in tracheal ELF ascorbic acid correlated with the percentage decrease in bronchoalveolar ELF ascorbic acid (r = 0.76; P = 0.004) following exposure. CONCLUSIONS: Acute organic dust exposure results in significant antioxidant depletion in the trachea, which may reflect inflammation and oxidative processes in peripheral airways. POTENTIAL RELEVANCE: Further work is required to evaluate the role of ascorbic acid depletion in the pathogenesis of RAO.

Changes in selected physiological and laboratory measurements in elite horses competing in a 160 km endurance ride.

REASON FOR PERFORMING STUDY: Limited information exists about the physiological changes and clinical problems that occur in elite horses competing in high-speed 160 km endurance races. OBJECTIVES: To provide initial data describing changes in physiological and laboratory measurements in horses competing in a high-speed, 160 km endurance race under temperate conditions and to compare data between horses that successfully completed the race and those that failed to finish. METHODS: Body mass (BM) was measured, blood samples were collected, and veterinary examinations performed on horses before, during, and at the finish of a CEI*** 160 km endurance race. RESULTS: Of 36 horses participating in the study, 22 (61%) completed the race. Twelve horses were eliminated for lameness and 2 for persistent heart rate elevation. Mean speed of finishers was 15.2 km/h. Mean +/- s.d. BM loss of finishers at the end of the race (5.7 +/- 2.6%) was not different (P = 0.58) from BM loss of nonfinishers at elimination (6.7 +/- 34%). Similarly, there were no significant differences in heart rate or veterinary assessment of hydration at the race end for finishers as compared to the elimination point for nonfinishers. PCV increased while sodium, chloride and potassium concentrations decreased with exercise but differences between finishers and nonfinishers were not detected. In contrast, both total and ionised calcium concentrations decreased in successful horses but remained unchanged in nonfinishers. CONCLUSIONS: Elite endurance horses are more likely to be eliminated from competition for lameness than metabolic problems; however, it remains unclear whether these conditions are entirely distinct. The magnitude of the decrease in sodium concentration in both finishers and nonfinishers was greater than in previous reports of 160 km rides. POTENTIAL RELEVANCE: These data should be of use for both organisers and participants in elite 160 km endurance races. The tendency toward hyponatraemia as well as the difference in calcium concentrations between finishers and nonfinishers warrant further study.

Retrospective study of predictive variables for maximal heart rate (HRmax) in horses undergoing strenuous treadmill exercise.

REASONS FOR PERFORMING STUDY: Heart rate is one of the most commonly measured variables in equine exercise physiology and relative exercise intensity commonly expressed as % of maximal heart rate. A number of influences affect maximal heart rate (HRmax), including age of the horse but other factors have not been described. OBJECTIVES: To determine if fitness, health status, gender, breed, athletic use, body mass, in addition to age, are predictive of HRmax in the horse. METHODS: Maximal heart rate data from 328 horses which underwent treadmill exercise tests at 5 different laboratories were obtained retrospectively. Univariable linear regression analyses were performed on individual variables. Multiple linear regression analysis using a backward elimination modelling procedure was then used to relate the observed HRmax values simultaneously with different predictive variables. Variables were retained in the final regression model if they or any of their categories were significantly predictive of HRmax at P<0.05 and if there was a significant collective contribution to the model from inclusion of each variable, also at P<0.05. RESULTS: Age, fitness status, laboratory, gender and breed/use (combined category) were all statistically significantly predictive of HRmax. Together these variables accounted for 41% of the variance in HRmax. Age alone accounted for only approximately 13% of the variation between horses in HRmax. Neither body mass nor health status were significantly predictive. CONCLUSIONS: HRmax in the horse declines with age but is also influenced by other factors. As the factors investigated accounted for only 41% of the variation between horses, other unidentified variables with a strong influence on HRmax remain to be identified. POTENTIAL RELEVANCE: Factors such as fitness, age, gender, breed and use need to be considered when interpreting estimates or measurements of HRmax.

Cardiac arrhythmias during and after treadmill exercise in poorly performing thoroughbred racehorses.

REASONS FOR PERFORMING STUDY: The prevalence and severity of cardiac arrhythmias during exercise in athletic horses presented for poor performance is not well described. OBJECTIVES: To describe prevalence and severity of ventricular and supraventricular arrhythmias immediately before, during and immediately after standardised incremental treadmill exercise tests (IET) to fatigue in Thoroughbred horses during investigation of poor performance. METHODS: The electrocardiograms (ECG) of 88 Thoroughbred racehorses, judged to be free of significant heart disease or arrhythmia at rest, were used. A modified base-apex ECG was recorded throughout an IET to fatigue. Recordings were analysed independently by 2 observers. Twenty-eight horses were diagnosed with dorsal displacement of the soft palate, 25 had varying degrees of soft palate instability and aryepiglottic fold collapse, 8 had other respiratory problems and, in 27 cases, no definitive diagnosis was reached. RESULTS: Fifty-five horses had at least one ventricular (VPD) or supraventricular (SVPD) depolarisation, 23 had only VPDs, 17 had only SVPDs and 15 had both in at least one exercise period. Premature depolarisations were seen predominantly during the first min of recovery from IET. The range of premature beats after exercise was 1-30 VPDs, and 1-9 SVPDs. No significant associations were observed between age, sex, race type, diagnosis, peak heart rate or run time to fatigue during IET and occurrence of either > or =1 premature beat or of more severe arrhythmias (multiple singles [>5] or pairs or paroxysms of premature depolarisations during peak exercise or immediately after exercise). However, a larger sample size would be required to have greater confidence in these associations. CONCLUSIONS: Isolated VPDs and SVPDs are frequently detected in poor performing racehorses during IET but their clinical relevance remains to be determined. POTENTIAL RELEVANCE: The guidelines for interpretation and clinical relevance of premature depolarisations observed during and immediately after treadmill exercise tests in poor performing Thoroughbred racehorses deserves further evaluation.

Effects of a specific endothelin-1A antagonist on exercise-induced pulmonary haemorrhage (EIPH) in thoroughbred horses.

REASONS FOR PERFORMING STUDY: During high intensity exercise, the very high pulmonary artery pressure (Ppa) experienced by Thoroughbred horses is considered a major factor in the aetiology of exercise-induced pulmonary haemorrhage (EIPH). Recently, endothelin-1 (ET-1), a potent vasoconstrictive hormone, has been found to increase Ppa in horses at rest via binding to its ET-1A receptor subtype. In addition, plasma concentrations of ET-1 are increased in horses during and after high intensity exercise. HYPOTHESIS: If ET-1 increases Ppa during exercise in the horse, administration of a specific ET-1A antagonist would decrease Ppa and therefore EIPH. METHODS: Saline (CON) or an ET-1A receptor antagonist, TBC3214 (3 mg/kg bwt i.v.; ANTAG) was administered to horses 1 h prior to maximal incremental exercise on a high-speed treadmill. Gas exchange measurements were made breath-by-breath and blood samples collected during each 1 min stage to determine blood gases, acid-base status and cardiac output. EIPH was determined via bronchoalveolar lavage (BAL) approximately 30 min after exercise. RESULTS: The time to fatigue, gas exchange and cardiovascular responses were not different between groups (P>0.05). Resting and peak Ppa did not differ significantly between treatments. Most importantly, ANTAG did not decrease EIPH. CONCLUSIONS: These results do not support a deterministic role for ET-1 in the increased Ppa and therefore EIPH, during maximal exercise in the equine athlete. POTENTIAL RELEVANCE: Treatment with an ET-1A receptor antagonist does not appear to be a viable therapeutic intervention in the prevention of EIPH.

The effect of inspired gas density on pulmonary artery transmural pressure and exercise induced pulmonary haemorrhage.

REASONS FOR PERFORMING STUDY: Pulmonary capillary stress failure, largely as a result of high pulmonary vascular pressures, has been implicated in the aetiology of EIPH. However, the role of the respiratory system in determining the magnitude of EIPH has received little attention. HYPOTHESIS: Horses breathing a gas of greater density than air will exhibit greater transmural pulmonary arterial pressures (TPAP) and more severe EIPH, and horses breathing a gas of lower density than air will exhibit lower TPAP and less severe EIPH, both compared with horses breathing air. METHODS: Following a warm-up, 8 Thoroughbred horses were exercised for 1 min at 10, 11 and 12 m/sec (5 degrees incline) breathing air or 21% oxygen/79% helium or 21% oxygen/79% argon in a randomised order. Heart rate, respiratory rate, pulmonary arterial pressure and oesophageal pressure were measured during exercise. Bronchoalveolar lavage fluid (BALF) was collected from the dorsocaudal regions of the left and right lungs 40 min post exercise and red blood cell (RBC) counts were performed. RESULTS: The exercise tests induced mild EIPH. Maximum changes in oesophageal pressure were lower on helium-oxygen compared to argon-oxygen (P<0.001). TPAP and median RBC counts did not differ between gas mixtures. BALF RBC counts from the left lung correlated with counts from the right lung (P<0.0001). However BALF RBC counts from the left lung were higher than those from the right lung (P = 0.004). CONCLUSION: As alterations in pulmonary arterial and oesophageal pressure caused by changes in inspired gas density were of similar magnitude, TPAP remained unchanged and there was no significant effect on EIPH severity. POTENTIAL RELEVANCE: Manipulations that decrease swings in intrapleural pressure may only decrease the degree of EIPH in horses severely affected by the condition.

Risk factors for epistaxis on British racecourses: evidence for locomotory impact-induced trauma contributing to the aetiology of exercise-induced pulmonary haemorrhage.

REASONS FOR PERFORMING STUDY: The proposed biological mechanisms for exercise-induced pulmonary haemorrhage (EIPH) are many and varied. Better knowledge of risk factors should lead to achievable measures to reduce the incidence. OBJECTIVES: To identify risk factors associated with epistaxis following racing in UK Thoroughbreds, to gain possible insights into the pathogenesis of the condition and to investigate the association between epistaxis and race finishing position. METHODS: The association of epistaxis occurring on UK racecourses between 1996 and 1998 with a wide range of race-, horse- and start-level variables was examined in multivariable mixed effect logistic regression analyses. Four multivariable analyses were conducted, one for all race types considered collectively and one each for flat, hurdle and steeplechase race types considered separately. RESULTS: Risk of epistaxis was significantly increased for hurdle and steeplechase race types compared to both flat and National Hunt flat races. In 3 of the 4 final models, there was a significant biological trend for increasing risk of epistaxis with increasing ground hardness ('going') and accumulated years spent racing. However, in flat races epistaxis was such a rare outcome (0.33 cases per 1000 starts) that this subset analysis had insufficient power to measure the detectable effect of 'going' as statistically significant. Horses with epistaxis were significantly more likely to have a poorer finishing position than those without blood at the nostrils. CONCLUSIONS: Findings were consistent with the theory that locomotory impact-induced trauma contributes to exercise-induced epistaxis. Further validation of this hypothesis through application of similar methods to endoscopically visible EIPH and through biomechanical studies is warranted. POTENTIAL RELEVANCE: Knowledge of significant risk factors should allow formulation of practical measures, such as track watering, to reduce the risks of EIPH and epistaxis in racehorses.

Antioxidant and inflammatory responses of healthy horses and horses affected by recurrent airway obstruction to inhaled ozone.

REASONS FOR PERFORMING STUDY: Inhaled ozone can induce oxidative injury and airway inflammation. Horses affected by recurrent airway obstruction (RAO) have a decreased pulmonary antioxidant capacity, which may render them more susceptible to oxidative challenge. It is currently unknown whether RAO-affected horses are more susceptible to oxidative stress than those unaffected by RAO. OBJECTIVES: To determine whether ozone exposure induces greater oxidative stress and airway inflammation in RAO-affected horses in remission than in healthy horses. METHODS: Seven healthy control horses and 7 RAO-affected horses were exposed to 0.8 ppm ozone for 2 h at rest. RESULTS: At baseline, bronchoalveolar lavage fluid (BALF) ascorbic acid concentrations were lower in RAO-affected horses than healthy controls. Ozone appeared to preferentially oxidise glutathione rather than ascorbic acid 6 h after exposure. Individual healthy and RAO-affected horses demonstrated oxidation of BALF glutathione after ozone exposure. Overall, RAO-affected horses did not demonstrate increased oxidative stress following ozone exposure, compared with healthy horses. Ozone did not induce significant airway inflammation in either group. CONCLUSIONS: RAO-affected horses in remission are not more sensitive to ozone despite a decreased pulmonary antioxidant capacity. Sensitivity to ozone appears to be independent of initial pulmonary antioxidant status. POTENTIAL RELEVANCE: Horses with high susceptibility to oxidative stress may benefit from antioxidant supplementation.

Use of an electronic nose to diagnose Mycobacterium bovis infection in badgers and cattle.

It is estimated that more than 50 million cattle are infected with Mycobacterium bovis worldwide, resulting in severe economic losses. Current diagnosis of tuberculosis (TB) in cattle relies on tuberculin skin testing, and when combined with the slaughter of test-positive animals, it has significantly reduced the incidence of bovine TB. The failure to eradicate bovine TB in Great Britain has been attributed in part to a reservoir of the infection in badgers (Meles meles). Accurate and reliable diagnosis of infection is the cornerstone of TB control. Bacteriological diagnosis has these characteristics, but only with samples collected postmortem. Unlike significant wild animal reservoirs of M. bovis that are considered pests in other countries, such as the brushtail possum (Trichosurus vulpecula) in New Zealand, the badger and its sett are protected under United Kingdom legislation (The Protection of Badgers Act 1992). Therefore, an accurate in vitro test for badgers is needed urgently to determine the extent of the reservoir of infection cheaply and without destroying badgers. For cattle, a rapid on-farm test to complement the existing tests (the skin test and gamma interferon assay) would be highly desirable. To this end, we have investigated the potential of an electronic nose (EN) to diagnose infection of cattle or badgers with M. bovis, using a serum sample. Samples were obtained from both experimentally infected badgers and cattle, as well as naturally infected badgers. Without exception, the EN was able to discriminate infected animals from controls as early as 3 weeks after infection with M. bovis, the earliest time point examined postchallenge. The EN approach described here is a straightforward alternative to conventional methods of TB diagnosis, and it offers considerable potential as a sensitive, rapid, and cost-effective means of diagnosing M. bovis infection in cattle and badgers.

Recording of ECG signals on a portable MiniDisc recorder for time and frequency domain heart rate variability analysis.

Analysis of heart rate variability (HRV) is a non-invasive technique useful for investigating autonomic function in both humans and animals. It has been used for research into both behaviour and physiology. Commercial systems for human HRV analysis are expensive and may not have sufficient flexibility for appropriate analysis in animals. Some heart rate monitors have the facility to provide inter-beat interval (IBI), but verification following collection is not possible as only IBIs are recorded, and not the raw electrocardiogram (ECG) signal. Computer-based data acquisition and analysis systems such as Po-Ne-Mah and Biopac offer greater flexibility and control but have limited portability. Many laboratories and veterinary surgeons have access to ECG machines but do not have equipment to record ECG signals for further analysis. The aim of the present study was to determine whether suitable HRV data could be obtained from ECG signals recorded onto a MiniDisc (MD) and subsequently digitised and analysed using a commercial data acquisition and analysis package. ECG signals were obtained from six Thoroughbred horses by telemetry. A split BNC connecter was used to allow simultaneous digitisation of analogue output from the ECG receiver unit by a computerised data acquisition system (Po-Ne-Mah) and MiniDisc player (MZ-N710, Sony). Following recording, data were played back from the MiniDisc into the same input channel of the data acquisition system as previously used to record the direct ECG. All data were digitised at a sampling rate of 500 Hz. IBI data were analysed in both time and frequency domains and comparisons between direct recorded and MiniDisc data were made using Bland-Altman analysis. Despite some changes in ECG morphology due to loss of low frequency content (primarily below 5 Hz) following MiniDisc recording, there was minimal difference in IBI or time or frequency domain analysis between the two recording methods. The MiniDisc offers a cost-effective approach to intermediate recording of ECG signals for subsequent HRV analysis and also provides greater flexibility than use of human Holter systems.

Validation of a cardiac monitor for measuring heart rate variability in adult female pigs: accuracy, artefacts and editing.

Autonomic regulation of cardiac activity during stress has not been clearly defined in farm animals. In part, this is due to the limited availability of affordable ambulatory cardiac monitors capable of accurately monitoring and storing large amounts of data that meet the criteria necessary for heart rate variability analysis. Our objectives were to measure the accuracy of a 24-h Polar RR monitor using gold standard ECG, to examine and categorise any occurring anomalies and to ascertain their impact on the outcome of heart rate variability analysis. Five 1-year-old female pigs (gilts) were socially isolated from their pen mates and cardiac activity was simultaneously measured using two systems, a 24-h Polar RR Recorder and a Telemetric ECG system. The Polar data were manually assessed both against and in isolation of the ECG data to identify anomalous beats, which were then assigned to one of five identified error categories. The anomalies in the Polar data were corrected and statistical comparisons were performed among the three data sets to evaluate the effects of anomalies on heart rate variability analysis. Bland-Altman analysis was used to measure the level of agreement among the ECG, Uncorrected Polar and Corrected Polar data. No anomalies or ectopies were found in the ECG data but 46 anomalies (0.81% of total interbeat intervals [IBI]) were found in the Polar Uncorrected data. Manual identification and editing procedures reduced this error to 0.018%. Most mean heart rate and IBI parameters were unaffected by error (P>.05). Standard deviation (S.D.) and root mean square of successive differences (RMSSD) were 45% and 50% higher when anomalies were present in the data. Artefacts affected the magnitude of the frequency domain indices and overestimated total and parasympathetic activity and underestimated sympathetic activity. The mean difference between ECG and Uncorrected Polar data was 1.36 ms (limits of agreement -69.03 to 71.74 ms). This was greatly improved to 0.36 ms (limits of agreement -5.37 to 6.10 ms) after editing. Overall, even a small proportion of error biased the outcome of heart rate variability analysis. This bias was greatly reduced by correcting the anomalous beats. Bland-Altman analysis demonstrated that when there was error present in the Polar data, it could not be used interchangeably with the ECG data. However, if there were no anomalies present in the data or if they were classified and corrected using the approach in this study, then the two systems could be used interchangeably.

Spasmogenic action of endothelin-1 on isolated equine pulmonary artery and bronchus.

REASONS FOR PERFORMING STUDY: There is currently little published information about the effects of endothelin-1 (ET-1), a potent endogenous spasmogen of vascular and airway smooth muscle, on pulmonary vasculature and airways or which ET receptor subtypes mediate ET-1-induced vasoconstrictive and bronchoconstrictive action in the horse. OBJECTIVES: To investigate the effect of endothelin-1 (ET-1) on smooth muscle from isolated equine pulmonary artery and bronchus. In addition, the roles of ETA and ETB receptors in ET-1 mediated contraction in these tissues were assessed. METHODS: The force generation of ring segments from pulmonary arteries or third-generation airways (obtained from horses subjected to euthanasia for orthopaedic reasons) were studied in an organ bath at 37 degrees C in response to exogenous endothelin and selective endothelin A (BQ123) or B receptor (BQ788) antagonists. RESULTS: ET-1 produced concentration-dependent contractions of the equine pulmonary artery and bronchus. The threshold for contraction was 10(-10) and 10(-9) mol/l ET-1 for pulmonary artery and bronchus, respectively. The maximal contraction induced by the highest ET-1 concentration (10(-7) mol/l) was 173 and 194% of the contraction obtained with 100 mmol/l KCl in pulmonary artery and bronchus, respectively. ET-1 potency was 25 times greater in equine pulmonary artery than in equine bronchus (concentration of ET-1 producing 50% of maximal contraction [EC50] = 5.6 10(-9) mol/l and 2.2 10(-8) mol/l, respectively). In pulmonary artery, ET-1 induced contractions were significantly inhibited by the ETA receptor antagonist BQ123 (1 micromol/l; dose-response curve to ET-1 was shifted to the right by 5.4-fold), but not by the ETB antagonist BQ788. In bronchus, dose-responses curves to ET-1 were shifted to the right by BQ123 (1 micromol/l; 2.5-fold), but not by BQ788 (1 micromol/l). In the presence of both antagonists, the dose-response curve to ET-1 was shifted to the right by 4.5-fold. CONCLUSIONS: These functional studies demonstrate that ET-1 is a potent spasmogen of equine third generation pulmonary artery and bronchus, and that contractions are mediated via ETA receptors in the former and both ETA and ETB receptors in the latter. POTENTIAL CLINICAL RELEVANCE: Endothelin receptor antagonists may have potential for treating equine pulmonary hypertension or bronchoconstriction.

Antioxidant supplementation and pulmonary function at rest and exercise.

Antioxidants have been implicated in the reduction and prevention of oxidative stress during exercise. We hypothesised that a dietary supplement containing a mixture of natural antioxidants together with vitamins E, C and selenium, given for 4 weeks, would increase the systemic and pulmonary antioxidant capacity leading to a reduction in markers of oxidative damage and an improvement in pulmonary function during exercise. In 6 healthy horses studied, the antioxidant supplement significantly increased plasma concentrations of ascorbic acid (from mean +/- s.d. 16 +/- 7 to 23 +/- 4 micromol/l; P = 0.007) and alpha-tocopherol (from 10 +/- 3 to 14 +/- 3 micromol/l; P = 0.02) and increased the bronchoalveolar lavage pulmonary epithelial lining fluid (ELF) concentration of ascorbic acid compared to a placebo, but not significantly (2.0 +/- 0.9 mmol/l and 1.2 +/- 0.9 mmol/l, respectively; P>0.05). Alpha-tocopherol was not detected in ELF either before or after supplementation or exercise. The mean concentration of malondialdehyde (MDA) in ELF was lower following antioxidant supplementation compared to placebo and control periods, but not significantly. An intermittent exercise test consisting of 2 min at 70, 80 and 90% of the horses' individual maximum oxygen uptake, failed to induce significant systemic or pulmonary oxidative stress (based on the glutathione redox ratio (GRR) and the ascorbic acid redox ratio (ARR)) and lipid peroxidation (based on the concentration of thiobarbituric acid reactive substances in plasma and MDA in ELF) either for placebo or antioxidant treatments. There was a strong correlation between GRR and ARR in the pulmonary epithelial lining fluid (r = 0.89; P<0.0001). In healthy horses on a diet containing adequate levels of antioxidants, additional antioxidant supplementation has no apparent beneficial or detrimental effect on pulmonary function during moderate intensity exercise. The importance of antioxidant supplementation may only become apparent if the diet is deficient in antioxidants, if exercise intensity is higher or more prolonged, or if disease or additional stresses are present.

Effect of dietary lipid on response to exercise: relationship to metabolic adaptation.

The aim of the present study was to relate changes in muscle oxidative capacity and free fatty acid flux in response to oil supplementation to fuel utilisation during subsequent exercise of varying intensities. Following 10 weeks of oil supplementation there was an increased capacity for fat utilisation during low and moderate intensity exercise as indicated by a lower respiratory exchange ratio (RER) (P<0.05). We suggest that this was contributed to by a parallel increase in the oxidative capacity of muscle as indicated by a significant increase in the activity of muscle citrate synthase (CS) (P<0.05) and trend towards an increase in beta-Hydroxy acyl CoA dehydrogenase (beta-HAD), (P>0.05). In addition, low and moderate intensity exercise was associated with an exercise-induced increase in plasma free fatty acids (FFA) and there was an increased facility for uptake of FFA by working muscle from circulating triglycerides, as suggested by an increase in TL activity (P<0.01). The response to oil supplementation varied between individual horses and the magnitude of response, during the low intensity exercise test, in terms of difference in RER was correlated to the increase in CS activity (r2 = 0.95, P<0.05) following oil supplementation. There was no similar significant correlation with respect to FFA, TL or beta-HAD activity (P>0.05). The hypothesis in this study was that the metabolic adaptation to oil supplementation, in terms of exercise response, was related to individual increases in the activities of CS, beta-HAD or TL. However, the relationship between these parameters was unequivocal and requires further investigation, ideally with a larger group of horses.

Movements of thoracic and abdominal compartments during ventilation at rest and during exercise.

The present investigation utilised simultaneous measurements of chest (Ch) and abdominal (Ab) circumferences and respiratory airflow to test the hypothesis that Ch circumferential expansion contributes proportionally little to tidal volume in the running Thoroughbred. During exercise, there were only small changes in Ch and Ab circumference and no increase with increasing tidal volume. At rest, walk and trot, the flow, Ch and Ab signals were in phase. However, during canter and gallop, the Ch and Ab changes were 180 degrees out of phase with each other and both were out of phase with airflow. In contrast to exercise, increase in ventilation at rest achieved by administration of lobeline resulted in a 4-6-fold increase in tidal volume; large excursions of the chest were always in phase with airflow. Furthermore, 3 horses showed an increase in chest circumference, demonstrating that chest stiffness per se does not preclude chest circumferential expansion. In conclusion, in the absence of significant increases in either Ch or Ab expansion during running, elongation of the thoracoabdominal segment may be the main determinant of tidal volume.

Modelling the oxygen cost of transport in competitions over ground of variable slope.

This study provides an objective method for estimating the oxygen consumption of horses while running on variable slopes so that realistic comparisons may be made of the locomotory transport cost involved in 3-day events, particularly the Speed and Endurance Test, at sites of differing terrain. A knowledge of the work profile over a particular course would enable competitors to plan speed and interval times appropriately along its length. We have developed a semi-empirical, but mechanistically based, model to calculate the oxygen cost of transport [COTpath in ml O2/kg/m path] for running on the flat, up or down a slope of given gradient (from -0.3 to +0.3). The model is then used to calculate the overall effort of running on a number of 3-day event courses of differing standard; the model does not assess the energetic cost of jumping. The cost of transport over the range of gradient of -0.3 to +0.3 was modelled using the following equations: On the flat or uphill: COTpath = 0.123 + 1.561(gradient); Downhill: COTpath = 0.123 + 1.591(gradient) + 9.762(gradient)2 + 14.0(gradient)3.

Airway cooling and mucosal injury during cold weather exercise.

In human subjects that exercise strenuously in cold weather, there is evidence that hyperventilation with cold air leads to peripheral airway cooling, desiccation and mucosal injury. Our hypothesis was that hyperventilation with cold air can result in penetration of unconditioned air (air that is not completely warmed and humidified) into the peripheral airways of exercising horses, resulting in peripheral airway mucosal injury. To test this hypothesis, a thermister-tipped catheter was inserted through the midcervical trachea and advanced into a sublobar bronchus in three horses that cantered on a treadmill at 6.6 m/s while breathing cold (5 degrees C) air. The mean (+/- s.e.) intra-airway temperature during cantering was 33.3 +/- 0.4 degrees C, a value comparable to the bronchial lumen temperatures measured in man during maximal exercise while breathing subfreezing dry air. In a second experiment, 6 fit Thoroughbred racehorses with satisfactory performance were used to determine whether strenuous exercise in cold conditions can produce airway injury. Horses were assigned to Exercise (E) or Control (C) groups in a random crossover design. Samples of bronchoalveolar lavage fluid (BALF) in the E treatment were recovered within 30 min of galloping exercise in 4 degrees C, 100% relative humidity (E), while in C BALF samples were obtained when the horses had not performed any exercise for at least 48 h prior. Ciliated epithelial cells in BALF were higher in E than in the C treatment. Similar results have been found in human athletes and laboratory animal models of cold weather exercise. These results support the hypothesis that, similar to man, horses that exercise in cold weather experience peripheral airway mucosal injury due to the penetration of unconditioned air. Furthermore, these results suggest that airway cooling and desiccation may be a factor in airway inflammation commonly found in equine athletes.