Effects of erythropoietin on systemic hematocrit and oxygen transport in the splenectomized horse.

To test the hypotheses that erythropoietin (rhuEPO) treatment increases systemic hematocrit, maximal O2 uptake (VO2max, by elevated perfusive and diffusive O2 conductances) and performance five female horses (4-13 years) received 15 IU/kg rhuEPO (erythropoietin) three times per week for three weeks. These horses had been splenectomized over 1 year previously to avoid confounding effects from the mobilization of splenic red blood cell reserves. Each horse performed three maximal exercise tests (one per month) on an inclined (4°) treadmill to the limit of tolerance; two control trials and one following EPO treatment. Measurements of hemoglobin concentration ([Hb] and hematocrit), plasma and blood volume, VO2, cardiac output as well as arterial and mixed venous blood gases were made at rest and during maximal exercise. EPO increased resting [Hb] by 18% from 13.3 ± 0.6 to 15.7 ± 0.8 g/dL (mean ± SD) corresponding to an increased hematocrit from 36 ± 2 to 46 ± 2% concurrent with 23 and 10% reductions in plasma and blood volume, respectively (all P<0.05). EPO elevated VO2max by 20% from 25.7 ± 1.7 to 30.9 ± 3.4 L/min (P<0.05) via a 17% increase in arterial O2 content and 18% greater arteriovenous O2 difference in the face of an unchanged cardiac output. To achieve the greater VO2max after EPO, diffusive O2 conductance increased ∼ 30% (from 580 ± 76 to 752 ± 166 mL O2/mmHg/min, P<0.05) which was substantially greater than the elevation of perfusive O2 conductance. These effects of EPO were associated with an increased exercise performance (total running time: control, 216 ± 72; EPO, 264 ± 48 s, P<0.05). We conclude that EPO substantially increases VO2max and performance in the splenectomized horse via improved perfusive and diffusive O2 transport.

Clinicopathologic evidence of myocardial injury in horses with acute abdominal disease.

OBJECTIVE: To determine whether there is evidence of myocardial injury in horses with acute abdominal disease. DESIGN: Prospective case series. ANIMALS: 18 healthy horses and 69 horses with acute abdominal disease. PROCEDURES: 18 healthy horses had been admitted to the hospital for investigation and were assigned to group 1. Horses examined for acute abdominal disease were assigned to 3 groups: strangulating obstruction, nonstrangulating obstruction, or inflammatory disease (groups 2, 3, and 4, respectively). Heart rate, Hct, and blood lactate and cardiac troponin I (cTnI) concentrations were measured at initial examination. Myocardial function was assessed by echocardiographic measurement of fractional shortening and left ventricular ejection time (LVET). Heart rhythm was evaluated via ECG. RESULTS: The proportion of horses with high (> 0.03 ng/mL) cTnI concentration was significantly greater among horses with strangulating (9/25 [36%]) or inflammatory (9/19 [47%]) lesions, compared with healthy horses (0/18). The proportion of horses with high cTnI concentration was significantly greater among nonsurvivors (12/24 [50%]) than among survivors (10/45 [22%]). Serum cTnI concentration was positively correlated with Hct, heart rate, and blood lactate concentration and negatively correlated with LVET. CONCLUSIONS AND CLINICAL RELEVANCE: Evidence of myocardial injury was observed in horses with acute abdominal disease, and this injury was associated with severity of illness. Recognition of myocardial injury could improve treatment of acute abdominal disease in horses.

Efficacy of furosemide for prevention of exercise-induced pulmonary hemorrhage in Thoroughbred racehorses.

OBJECTIVE: To evaluate the efficacy of furosemide for prevention of exercise-induced pulmonary hemorrhage (EIPH) in Thoroughbred racehorses under typical racing conditions. DESIGN: Randomized, placebo-controlled, blinded, crossover field trial. ANIMALS: 167 Thoroughbred racehorses. PROCEDURES: Horses were allocated to race fields of 9 to 16 horses each and raced twice, 1 week apart, with each of the 2 races consisting of the same race field and distance. Each horse received furosemide (500 mg, IV) before one race and a placebo (saline solution) before the other, with the order of treatments randomly determined. Severity of EIPH was scored on a scale from 0 to 4 after each race by means of tracheobronchoscopy. Data were analyzed by means of various methods of multivariable logistic regression. RESULTS: Horses were substantially more likely to develop EIPH (severity score >or= 1; odds ratio, 3.3 to 4.4) or moderate to severe EIPH (severity score >or= 2; odds ratio, 6.9 to 11.0) following administration of saline solution than following administration of furosemide. In addition, 81 of the 120 (67.5%) horses that had EIPH after administration of saline solution had a reduction in EIPH severity score of at least 1 when treated with furosemide. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that prerace administration of furosemide decreased the incidence and severity of EIPH in Thoroughbreds racing under typical conditions in South Africa.

Temporal pattern of skeletal muscle gene expression following endurance exercise in Alaskan sled dogs.

Muscle responses to exercise are complex and include acute responses to exercise-induced injury, as well as longer term adaptive training responses. Using Alaskan sled dogs as an experimental model, changes in muscle gene expression were analyzed to test the hypotheses that important regulatory elements of the muscle's adaptation to exercise could be identified based on the temporal pattern of gene expression. Dogs were randomly assigned to undertake a 160-km run (n=9), or to remain at rest (n=4). Biceps femoris muscle was obtained from the unexercised dogs and two dogs at each of 2, 6, and 12 h after the exercise, and from three dogs 24 h after exercise. RNA was extracted and microarray analysis used to define gene transcriptional changes. The changes in gene expression after exercise occurred in a temporal pattern. Overall, 569, 469, 316, and 223 transcripts were differentially expressed at 2, 6, 12, and 24 h postexercise, respectively, compared with unexercised dogs (based on Por=1.5). Increases in a number of known transcriptional regulators, including peroxisome proliferator-activated receptor-alpha, cAMP-responsive element modulator, and CCAAT enhancer binding protein-delta, and potential signaling molecules, including brain-derived neurotrophic factor, dermokine, and suprabasin, were observed 2 h after exercise. Biological functional analysis suggested changes in expression of genes with known functional relationships, including genes involved in muscle remodeling and growth, intermediary metabolism, and immune regulation. Sustained endurance exercise by Alaskan sled dogs induces coordinated changes in gene expression with a clear temporal pattern. RNA expression profiling has the potential to identify novel regulatory mechanisms and responses to exercise stimuli.

Assessment of alterations in triglyceride and glycogen concentrations in muscle tissue of Alaskan sled dogs during repetitive prolonged exercise.

OBJECTIVE: To assess changes in muscle glycogen (MG) and triglyceride (MT) concentrations in aerobically conditioned sled dogs during prolonged exercise. ANIMALS: 54 Alaskan sled dogs fed a high-fat diet. PROCEDURES: 48 dogs ran 140-km distances on 4 consecutive days (cumulative distance, up to 560 km); 6 dogs remained as nonexercising control animals. Muscle biopsies were performed immediately after running 140, 420, or 560 km (6 dogs each) and subsequently after feeding and 7 hours of rest. Single muscle biopsies were performed during recovery at 28 hours in 7 dogs that completed 560 km and at 50 and 98 hours in 7 and 6 dogs that completed 510 km, respectively. Tissue samples were analyzed for MG and MT concentrations. RESULTS: In control dogs, mean +/- SD MG and MT concentrations were 375 +/- 37 mmol/kg of dry weight (kgDW) and 25.9 +/- 10.3 mmol/kgDW, respectively. Compared with control values, MG concentration was lower after dogs completed 140 and 420 km (137 +/- 36 mmol/kgDW and 203 +/- 30 mmol/kgDW, respectively); MT concentration was lower after dogs completed 140, 420, and 560 km (7.4 +/- 5.4 mmol/kgDW; 9.6 +/- 6.9 mmol/kgDW, and 6.3 +/- 4.9 mmol/kgDW, respectively). Depletion rates during the first run exceeded rates during the final run. Replenishment rates during recovery periods were not different, regardless of distance; only MG concentration at 50 hours was significantly greater than the control value. CONCLUSIONS AND CLINICAL RELEVANCE: Concentration of MG progressively increased in sled dogs undergoing prolonged exercise as a result of attenuated depletion.

Assessment of serum antibody titers against canine distemper virus, canine adenovirus type II, and canine parvovirus in Alaskan sled dogs before and after a long-distance race.

OBJECTIVE: To determine serum antibody titers against canine distemper virus (CDV), canine adenovirus type II (CAV-2), and canine parvovirus (CPV) in trained sled dogs prior to and after completion of a long-distance race. DESIGN: Prospective cohort study. ANIMALS: 195 Alaskan sled dogs (from 18 kennels) that participated in the 2006 Iditarod Trail Race. PROCEDURES: All 1,323 dogs participating in the race had been vaccinated against the 3 viruses at 19 to 286 days prior to initial blood sample collection (obtained within the month preceding the race). Within 12 hours of race completion, blood samples were collected from 195 dogs (convenience sample) and matched with each dog's prerace sample. Serum antibody titers (90% confidence intervals [CIs]) were determined via serum neutralization assays. RESULTS: After racing, geometric mean titers against CDV and CPV were significantly higher (2,495 [90% CI, 321 to 16,384] and 6,323 [90% CI, 512 to 32,768], respectively) than prerace values (82 [90% CI, 11 to 362] and 166 [90% CI, 32 to 1,024], respectively). Sixty-one of 194 (31.4%) dogs had > or = 4-fold increases in anti-CPV antibody titers after racing. Prerace serum antibody titers against CDV, CPV, and CAV-2 varied significantly by sled team but were not associated with time since vaccination. CONCLUSIONS AND CLINICAL RELEVANCE: Postrace increases in serum anti-CDV and anti-CPV antibody titer might reflect exposure of dogs to these agents immediately before or during racing. Dogs had no clinical signs of CDV-, CAV-2-, or CPV-associated disease; therefore, the clinical importance of these titer changes is uncertain.

Acid-base and hormonal abnormalities in dogs with naturally occurring diabetes mellitus.

OBJECTIVE: To examine acid-base and hormonal abnormalities in dogs with diabetes mellitus. DESIGN: Cross-sectional study. ANIMALS: 48 dogs with diabetes mellitus and 17 healthy dogs. PROCEDURES: Blood was collected and serum ketone, glucose, lactate, electrolytes, insulin, glucagon, cortisol, epinephrine, norepinephrine, nonesterified fatty acid, and triglyceride concentrations were measured. Indicators of acid-base status were calculated and compared between groups. RESULTS: Serum ketone and glucose concentrations were significantly higher in diabetic than in healthy dogs, but there was no difference in venous blood pH or base excess between groups. Anion gap and strong ion difference were significantly higher and strong ion gap and serum bicarbonate concentration were significantly lower in the diabetic dogs. There were significant linear relationships between measures of acid-base status and serum ketone concentration, but not between measures of acid-base status and serum lactate concentration. Serum insulin concentration did not differ significantly between groups, but diabetic dogs had a wider range of values. All diabetic dogs with a serum ketone concentration > 1,000 micromol/L had a serum insulin concentration < 5 microU/mL. There were strong relationships between serum ketone concentration and serum glucagon-insulin ratio, serum cortisol concentration, and plasma norepinephrine concentration. Serum beta-hydroxybutyrate concentration, expressed as a percentage of serum ketone concentration, decreased as serum ketone concentration increased. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that ketosis in diabetic dogs was related to the glucagon-insulin ratio with only low concentrations of insulin required to prevent ketosis. Acidosis in ketotic dogs was attributable largely to high serum ketone concentrations.

Effects of training and strenuous exercise on hematologic values and peripheral blood leukocyte subsets in racing sled dogs.

OBJECTIVE: To determine the effects of training and sustained submaximal exercise on hematologic values in racing sled dogs. DESIGN: Cohort study. ANIMALS: 39 Alaskan sled dogs bred for endurance racing. Procedures-Blood samples were collected prior to initiation of a 7-month training regimen (n=39), after completion of the training regimen (19), and after completion of an 1,100-mile race (9), and a CBC, differential cell count, and flow cytometry for leukocyte surface antigens were performed. RESULTS: Both training and exercise caused significant decreases in PCV and hemoglobin concentration and significant increases in total WBC count. In contrast, training and exercise were not found to have significant effects on absolute numbers or fractions of CD4+ or CD8+ lymphocytes, other than a significant increase in the fraction of CD8+ lymphocytes associated with training. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that training and exercise induced changes in several hematologic values in racing sled dogs. Extracellular fluid volume expansion was the likely explanation for the training-induced decrease in PCV, and acute blood loss secondary to gastrointestinal tract bleeding was likely responsible for the decrease in PCV associated with acute exercise.

Clinical and clinicopathologic variables in adult horses receiving blood transfusions: 31 cases (1999-2005).

OBJECTIVE: To determine clinical and clinicopathologic abnormalities in horses administered a blood transfusion and evaluate effects of blood transfusion on these variables. DESIGN: Retrospective case series. ANIMALS: 31 adult horses that received > or = 1 blood transfusion. Procedures-Medical records of horses receiving a blood transfusion were reviewed to obtain clinical findings, laboratory test results before and after transfusion, adjunctive treatments, transfusion type and volume, response to transfusion, results of donor-recipient compatibility testing, adverse reactions, and outcome. RESULTS: 31 horses received 44 transfusions for hemorrhagic anemia (HG; n = 18 horses), hemolytic anemia (HL; 8), or anemia attributable to erythropoietic failure (EF; 5). Tachycardia and tachypnea were detected in 31 of 31 (100%) and 22 of 31 (71%) horses, respectively, before transfusion. The PCV and hemoglobin concentration were less than the reference range in 11 of 18 horses with HG, 8 of 8 horses with HL, and 5 of 5 horses with EF. Hyperlactatemia was detected in 16 of 17 recorded values before transfusion. Heart rate, respiratory rate, and PCV improved after transfusion, with differences among the types of anemia. Seventeen (54%) horses were discharged, 9 (29%) were euthanized, and 5 (16%) died of natural causes. Adverse reactions were evident during 7 of 44 (16%) transfusions, varying from urticarial reactions to anaphylactic shock. CONCLUSIONS AND CLINICAL RELEVANCE: Abnormalities in clinical and clinicopathologic variables differed depending on the type of anemia. Colic, cold extremities, signs of depression, lethargy, tachycardia, tachypnea, low PCV, low hemoglobin concentration, and hyperlactatemia were commonly detected before transfusion and resolved after transfusion.

Serum chemistry alterations in Alaskan sled dogs during five successive days of prolonged endurance exercise.

OBJECTIVE: To determine the impact of successive days of endurance exercise on select serum chemistry values in conditioned Alaskan sled dogs. DESIGN: Prospective cohort study. ANIMALS: 10 conditioned Alaskan sled dogs. PROCEDURES: All dogs ran 160 km/d for 5 consecutive days. Serum was obtained prior to exercise and immediately after each exercise run; all samples were obtained before dogs were fed. Serum electrolyte, mineral, protein, total bilirubin, urea nitrogen, creatinine, and cardiac troponin-I concentrations and serum alkaline phosphatase, alanine aminotransferase, creatine kinase, and aspartate aminotransferase activities were measured. Data were analyzed by means of analysis of covariance for a randomized complete block design with dog as a blocking variable, time as a covariate, and distance run as the treatment of interest. Least square mean values were compared with values obtained prior to exercise, and linear and quadratic contrasts were examined. RESULTS: Serum globulin concentration was low prior to exercise (mean +/- SD, 2.2 +/- 0.3g/dL) and progressively decreased as exercise continued. Exercise was associated with increases in serum chloride, urea nitrogen, and cardiac troponin-I concentrations and serum alanine aminotransferase, creatine kinase, and aspartate aminotransferase activities and with progressive decreases in serum potassium, total protein, and albumin concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that multiple successive days of endurance exercise resulted in mild aberrations in serum chemistry variables in conditioned sled dogs. Changes likely reflected the metabolic stresses of prolonged endurance exercise as well as dietary composition. Hypoglobulinemia in resting, conditioned sled dogs may reflect the immunosuppressive or catabolic effects of intense endurance training.

Effect of strenuous exercise on urine concentrations of homovanillic acid, cortisol, and vanillylmandelic acid in sled dogs.

OBJECTIVE: To determine whether prolonged exercise by conditioned sled dogs affects urine concentrations of homovanillic acid (a metabolite of dopamine), vanillylmandelic acid (a metabolite of norepinephrine and epinephrine), and cortisol. ANIMALS: 24 conditioned Alaskan sled dogs (2 to 8.5 years old) that were in training for a multiday endurance race. PROCEDURES: Voided urine samples were collected from 4 groups of dogs (randomly selected from 54 dogs) after no exercise (control group; n = 6 dogs), completion of a 160km run (group A; 3), completion of a 420-km run (group B; 7), and completion of a 560-km run (group C; 6). Urine cortisol concentrations were determined by use of an immunoassay technique; urine vanillylmandelic acid and homovanillic acid concentrations were measured via high-performance liquid chromatography. RESULTS: Compared with the control group, urine cortisol concentration in groups A, B, and C was significantly different (5.33 x 10(4) +/- 2.62 x 10(4) microg/dL vs 1.04 x 10(4) +/- 2.31 x 10(5) microg/dL, 8.88 x 10(4) +/- 5.49 x 10(4) microg/dL, and 6.31 x 10(4) +/- 5.09 x 10(4) microg/dL, respectively). Urine homovanillic acid concentration did not differ among the 4 groups. Vanillylmandelic acid was not detected in any urine samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that prolonged exercise by sled dogs did not affect urine homovanillic acid concentration but did increase urinary cortisol secretion, which is indicative of adrenocortical stimulation. The apparent lack of vanillylmandelic acid in voided urine samples requires further investigation.

Usefulness of a commercial equine IgG test and serum protein concentration as indicators of failure of transfer of passive immunity in hospitalized foals.

Detection of failure of transfer of passive immunity (FTPI) is important in reducing morbidity and mortality in neonatal foals. We investigated the performance of a commercial equine IgG test (SNAP Foal IgG Test Kit) to diagnose FTPI in hospitalized foals. Furthermore, we evaluated the usefulness of serum total protein (STP) and serum globulin (SG) concentrations as indicators of FTPI. Serum IgG concentration was measured by means of the SNAP test and single radial immunodiffusion, and SG and STP concentrations were determined by means of a clinical chemistry analyzer. Subjects were 67 hospitalized foals <19 days old. The SNAP test was repeated on 37 samples from 29 foals, with identical results for 24 samples (kappa statistic, 0.64; 95% confidence interval [CI], 0.46-0.82). The sensitivity of the SNAP test to detect serum IgG concentration [IgG] < or =400 and < or =800 mg/dl was 90% (95% CI, 71-98%) and 95% (85-99%), respectively, and the specificity was 79% (71-82%) and 52% (39-57%), respectively. Sensitivity for detection of [IgG] < or =400 mg/dl was not affected (P > .05) by plasma fibrinogen concentration, sepsis score, or bacteremia. Specificity for detection of [IgG] < or = 800 mg/dl was lower (P < .05) in foals with sepsis score < or =11 (50% [31-60%] versus 100% [8-100%]) and bacteremia (25% [5-56%] versus 62% [45-62%]). Sensitivity and specificity of [STP] < or = 5.0 g/dl for [IgG] < or =800 mg/dl was 94% (83-99%) and 47% (30-56%), respectively. Performance of the SNAP test in hospitalized foals is impaired because of low specificity, but can have usefulness provided that the properties of the test and characteristics of the foal being examined are considered when interpreting the results. The STP and SG concentrations are poor sole indicators of FTPI in hospitalized foals, but may be useful adjunctive tests.

Ingestion of starch-rich meals after exercise increases glucose kinetics but fails to enhance muscle glycogen replenishment in horses.

Fatiguing exercise substantially decreases muscle glycogen concentration in horses, impairing athletic performance in subsequent exercise bouts. Our objective was to determine the effect of ingestion of starch-rich meals after exercise on whole body glucose kinetics and muscle glycogen replenishment. In a randomized, cross-over study seven horses with exercise-induced muscle glycogen depletion were either not fed for 8 h, fed half of the daily energy requirements ( approximately 15 Mcal DE) as hay, or fed an isocaloric amount of corn 15 min and 4 h after exercise. Starch-rich meals fed after exercise, when compared to feed withholding, resulted in mild to moderate hyperglycemia (5.7+/-0.3 vs. 4.7+/-0.3 mM, P<0.01) and hyperinsulinemia (79.9+/-9.3 vs. 39.0+/-1.9 pM, P<0.001), 3-fold greater whole body glucose kinetics (15.5+/-1.4 vs. 5.3+/-0.4 micromol kg(-1)min(-1), P<0.05), but these only minimally enhanced muscle glycogen replenishment (171+/-19 vs. 170+/-56 and 260+/-45 vs. 294+/-29 mmol/kg dry weight immediately and 24 h after exercise, P>0.05). It is concluded that after substantial exercise-induced muscle glycogen depletion, feeding status only minimally affects net muscle glycogen concentrations after exercise, despite marked differences in soluble carbohydrate ingestion and availability of glucose to skeletal muscle.

Association between exercise-induced pulmonary hemorrhage and performance in Thoroughbred racehorses.

OBJECTIVE: To determine whether exercise-induced pulmonary hemorrhage (EIPH) was associated with racing performance inThoroughbred horses not medicated with furosemide and not using nasal dilator strips. DESIGN: Observational cross-sectional study. ANIMALS: 744 two- to 10-year-old Thoroughbred horses racing in Melbourne, Australia. PROCEDURE: Horses were enrolled prior to racing, and a tracheobronchoscopic examination was performed after 1 race. Examinations were recorded on videotape, and presence and severity (grade 0 to 4) of EIPH were subsequently determined by 3 observers blinded to the horses' identity. Race records were abstracted for each horse examined. RESULTS: Overall, 52.1% of horses eligible for participation in the study were examined, and horses that were examined did not differ from horses that were not examined in regard to age, sex distribution, or proportion of horses that won or finished in the first 3 positions. Horses with EIPH grades < 1 were 4.0 times as likely to win, 1.8 times as likely to finish in the first 3 positions, and 3.03 times as likely to be in the 90th percentile or higher for race earnings as were horses with grades > 2. Horses with EIPH grades > 1 finished significantly farther behind the winner than did horses without EIPH. However, odds that horses with grade 1 EIPH would win or finish in the first 3 positions were not significantly different from odds for horses without EIPH. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that EIPH is associated with impaired performance in Thoroughbred racehorses not medicated with furosemide and not using nasal dilator strips.

Evaluation of plasma catecholamine and serum cortisol concentrations in horses with colic.

OBJECTIVE: To evaluate plasma epinephrine and norepinephrine concentrations and serum cortisol concentration in horses with colic and assess the relationship of these variables with clinical signs, routinely measured clinicopathologic variables, and outcome in affected horses. DESIGN: Prospective observational study. ANIMALS: 35 horses with colic. PROCEDURE: Blood samples were collected within 30 minutes of arrival at the veterinary hospital from horses referred because of colic. Plasma and serum samples were analyzed for cortisol, epinephrine, norepinephrine, lactate, and electrolyte concentrations and acid-base variables. Heart rate at admission and outcome (survival or nonsurvival) were recorded. Univariate logistic regression was used to calculate crude (unadjusted) odds ratios and 95% confidence intervals. RESULTS: Of the 35 horses with colic, 26 survived. Higher plasma epinephrine, plasma lactate, and serum cortisol concentrations were significantly associated with increased risk of nonsurvival, but plasma norepinephrine concentration was not associated with outcome. Plasma epinephrine concentration was significantly correlated with heart rate (r = 0.68), plasma lactate concentration (r = 0.87), blood pH (r = -0.83), anion gap (r = 0.74), and base excess (r = -0.81). CONCLUSIONS AND CLINICAL RELEVANCE: The risk of death appears to be greater in colic-affected horses with high circulating concentrations of epinephrine and cortisol. The correlation of epinephrine with other biochemical markers of illness severity and with heart rate indicates that the degree of sympathetic activation in horses with colic can be inferred from routinely measured variables.