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Objective: To compare, in horses, estimates of insulin sensitivity obtained from minimal model analysis (MMA) of a frequently sampled IV glucose tolerance test (FSIGTT) with estimates from the euglycemic-hyperinsulinemic clamp (EHC) and to evaluate the validity of surrogate estimates of insulin sensitivity derived from an oral glucose tolerance test (OGTT)., Animals: 18 mature Standardbreds (mean ± SD body weight, 428.9 ± 35.9 kg; mean ± SD body condition score, 4.4 ± 1.0 [on a scale of 1 to 9])., Procedures: All horses underwent at least 2 of the 3 procedures (EHC [n = 15], insulin-modified FSIGTT [18], and OGTT [18]) within a 10-day time frame to evaluate insulin sensitivity., Results: Insulin sensitivity variables derived from the EHC and FSIGTT were strongly correlated (r = 0.88). When standardized to the same units of measure, these measures were still strongly correlated (r = 0.86) but were not equivalent. Area under the curve, peak insulin concentration, insulin concentration at 120 minutes, and 2 calculated indices from glucose and insulin data from the OGTT were significantly correlated with the EHC- and FSIGTT-derived estimates of insulin sensitivity., Conclusions and Clinical Relevance: In healthy Standardbreds with moderate body condition score, insulin sensitivities from the EHC and FSIGTT were strongly correlated but not equivalent. Estimates derived from an OGTT also may be useful to estimate insulin sensitivity.

Objective: To characterize the effects of pregnancy on insulin sensitivity (SI) and glucose dynamics in pasture-maintained mares fed supplemental feeds of differing energy composition., Animals: Pregnant (n = 22) and nonpregnant (10) healthy Thoroughbred mares., Procedures: Pregnant and nonpregnant mares underwent frequently sampled intravenous glucose tolerance tests at 2 times (period 1, 25 to 31 weeks of gestation; period 2, 47 weeks of gestation). Following period 1 measurements, mares were provided a high-starch (HS; 39% starch) or high-fat and -fiber (14% fat and 70% fiber) supplemental feed. From a subset of mares (n = 12), blood samples were collected hourly for 24 hours to assess glycemic and insulinemic response to feeding while pastured. The minimal model of glucose and insulin dynamics was used to estimate SI, glucose effectiveness, and acute insulin response to glucose from tolerance testing data., Results: Pregnant mares during period 1 had a lower SI and glucose effectiveness and higher acute insulin response to glucose than did nonpregnant mares. The SI value decreased in nonpregnant but not pregnant mares from periods 1 to 2. Pregnant mares fed HS feed had a greater glycemic and insulinemic response to feeding than did any other group., Conclusions and Clinical Relevance: Pregnant mares had slower glucose clearance and greater insulin secretion at 28 weeks of gestation than did nonpregnant mares. Glucose and insulin responses to meal feeding, particularly with HS feed, were greater in pregnant mares, indicating that pregnancy enhanced the postprandial glycemic and insulinemic effects of starch-rich feed supplements.

Objective: To investigate the effects of a continuous rate infusion (CRI) of dextrose solution or dextrose solution and insulin on glucose and insulin concentrations in healthy and endotoxin-exposed horses., Animals: 9 adult mares., Procedures: During phase 1, treatments consisted of saline (0.9% NaCl) solution (control group; n = 4) or 20% dextrose solution (group 1; 4) administered IV as a 360-minute CRI. During phase 2, treatments consisted of 360-minute CRIs of 20% dextrose solution and insulin administered simultaneously at 367.6 mg/kg/h (30 kcal/kg/d) and 0.07 U/kg/h, respectively, in healthy horses (group 2; n = 4) or horses administered 35 ng of lipopolysaccharide/kg, IV, 24 hours before starting the dextrose solution and insulin CRIs (group 3; 4). A balanced crossover study design was used in both phases. Blood samples were collected for measurement of plasma glucose and insulin concentrations., Results: Infusion of dextrose solution alone resulted in hyperglycemia for most of the 360-minute CRI. Insulin concentration increased significantly in group 1, compared with that in the control group. Mean insulin concentration of group 2 was significantly higher throughout most of the infusion period, compared with concentrations of the control group and group 1. Mean glucose concentration did not differ significantly between groups 2 and 3., Conclusions and Clinical Relevance: Insulin infusion at a rate of 0.07 U/kg/h was found to be effective for the prevention of hyperglycemia when administered concurrently with dextrose solution. This rate was considered to be safe because horses did not become hypoglycemic during infusions of dextrose solution.

Objective: To determine the effect of refeeding following an 18-hour period of feed withholding on the phosphorylation of translation initiation factors in the skeletal muscle of mature horses., Animals: 8 adult horses., Procedures: Following an 18-hour period of feed withholding, horses either continued to have feed withheld (postabsorptive state) or were fed 2 g/kg of a high-protein feed (33% crude protein) at time 0 and 30 minutes (postprandial state). Blood samples were taken throughout the experimental period. At 90 minutes, a biopsy specimen was taken from the middle gluteal muscle to measure the phosphorylation of translation initiation factors and tissue amino acid concentrations. Plasma glucose, insulin, and amino acid concentrations were also measured., Results: Horses in the postprandial state had significantly higher plasma insulin, glucose, and amino acid concentrations than did those in the postabsorptive state at the time of biopsy. Refeeding significantly increased the phosphorylation state of riboprotein S6 and eukaryotic initiation factor 4E binding protein 1., Conclusions and Clinical Relevance: In mature horses, feeding resulted in increased mammalian target of rapamycin signaling and the mechanism appeared to be independent of an increase in Akt phosphorylation at Ser⁴⁷³. Results indicate that adult horses may be able to increase rates of muscle protein synthesis in response to feeding and that dietary amino acids appear to be the main mediators of this effect.

Objective: To determine effects of exercise training without dietary restriction on adiposity, basal hormone and lipid concentrations and glucose and insulin dynamics in overweight or obese, insulin-resistant horses., Animals: 12 overweight or obese (body condition score > or = 7), insulin-resistant (insulin sensitivity < or = 1.2 x 10(-4) L/min/mU) geldings., Procedures: 4 horses remained sedentary, and 8 horses were exercised for 4 weeks at low intensity and 4 weeks at higher intensity, followed by 2 weeks of detraining. Prior to and after each training period, frequently sampled IV glucose tolerance tests with minimal model analysis were performed and baseline plasma insulin, glucose, triglycerides, non-esterified fatty acids, and leptin concentrations were analyzed. Adiposity was assessed by use of morphometrics, ultrasonic subcutaneous fat thickness, and estimation of fat mass from total body water (deuterium dilution method)., Results: Body weight and fat mass decreased by 4% (mean +/- SD, 20 +/- 8 kg) and 34% (32 +/- 9 kg), respectively, compared with pre-exercise values, with similar losses during low- and higher-intensity training. There was no effect of exercise training on subcutaneous fat thickness, plasma hormone and lipid concentrations, or minimal model parameters of glucose and insulin dynamics., Conclusions and Clinical Relevance: Results suggested that moderate exercise training without concurrent dietary restriction does not mitigate insulin resistance in overweight or obese horses. A more pronounced reduction in adiposity or higher volume or intensity of exercise may be necessary for improvement in insulin sensitivity in such horses.

Objective: To investigate the effects of dexamethasone or levothyroxine sodium on endotoxin-induced alterations in glucose and insulin dynamics., Animals: 24 horses., Procedures: Horses were randomly allocated to 3 treatment groups and received 48 mg of levothyroxine mixed with 200 g of oats, 20 mg of dexamethasone plus oats, or oats alone (control) for 15 days, followed by IV infusion of lipopolysaccharide (20 ng/kg) while individually housed in stalls. Frequently sampled IV glucose tolerance tests were performed prior to pretreatment, after pretreatment, and 20 hours after lipopolysaccharide administration. Area under the curve for plasma glucose and serum insulin concentrations was calculated, and minimal model analyses were performed., Results: Significant treatment-by-time effects were detected for insulin sensitivity (SI) and area under the curve for glucose and insulin in the 15-day pretreatment period. Insulin sensitivity significantly decreased over time in all treatment groups, with the largest decrease detected in the dexamethasone group. Administration of lipopolysaccharide further decreased mean SI by 71% and 63% in the dexamethasone and control groups, respectively, but did not affect horses in the levothyroxine group. Mean SI was the lowest in the dexamethasone group, but percentage reduction was the same for dexamethasone and control groups., Conclusions and Clinical Relevance: Insulin sensitivity decreased during the pretreatment period in all 3 groups, indicating that hospitalization affected glucose and insulin dynamics. Dexamethasone significantly lowered SI, and endotoxemia further exacerbated insulin resistance. In contrast, there was no additional effect of endotoxemia on SI in horses pretreated with levothyroxine, suggesting that this treatment prevented endotoxemia-induced insulin resistance.

Objective: To determine the effects of diet-induced weight gain on glucose and insulin dynamics and plasma hormone and lipid concentrations in horses., Animals: 13 adult geldings., Procedures: Horses were fed 200% of their digestible energy requirements for maintenance for 16 weeks to induce weight gain. Frequently sampled IV glucose tolerance tests were performed before and after weight gain to evaluate glucose and insulin dynamics. Adiposity (assessed via condition scoring, morphometric measurements, and subcutaneous fat depth) and plasma concentrations of insulin, glucose, nonesterified fatty acids, triglycerides, and leptin were measured on a weekly or biweekly basis., Results: Mean + or - SD body weight increased by 20% from 440 + or - 44 kg to 526 + or - 53 kg, and body condition score (scale, 1 to 9) increased from 6 + or - 1 to 8 + or - 1. Plasma glucose, triglyceride, and nonesterified fatty acid concentrations were similar before and after weight gain. Leptin and insulin concentrations increased with weight gain. Mean + or - SD insulin sensitivity decreased by 71 + or - 28%, accompanied by a 408 + or - 201% increase in acute insulin response to glucose, which resulted in similar disposition index before and after weight gain., Conclusions and Clinical Relevance: Diet-induced weight gain in horses occurred concurrently with decreased insulin sensitivity that was effectively compensated for by an increase in insulin secretory response. Obesity resulted in hyperinsulinemia and hyperleptinemia, compared with baseline values, but no changes in lipid concentrations were apparent. Preventing obesity is a potential strategy to help avoid insulin resistance, hyperinsulinemia, and hyperleptinemia in horses.

Objective: To assess a technique for laparoscopic collection of serial full-thickness small intestinal biopsy specimens in horses., Animals: 13 healthy adult horses., Procedures: In the ex vivo portion of the study, sections of duodenum and jejunum obtained from 6 horses immediately after euthanasia were divided into 3 segments. Each segment was randomly assigned to the control group, the double-layer hand-sewn closure group, or the endoscopic linear stapler (ELS) group. Bursting strength and bursting wall tension were measured and compared among groups; luminal diameter reduction at the biopsy site was compared between the biopsy groups. In the in vivo portion of the study, serial full-thickness small intestinal biopsy specimens were laparoscopically collected with an ELS from the descending duodenum and distal portion of the jejunum at monthly intervals in 7 sedated, standing horses. Biopsy specimens were evaluated for suitability for histologic examination., Results: Mean bursting strength and bursting wall tension were significantly lower in the ELS group than in the hand-sewn and control groups in both the duodenal and jejunal segments. Use of the hand-sewn closure technique at the biopsy site reduced luminal diameter significantly more than use of the stapling technique. In the in vivo part of the study, all 52 biopsy specimens collected during 26 laparoscopic procedures were suitable for histologic examination and no clinically important perioperative complications developed., Conclusions and Clinical Relevance: Laparoscopic collection of serial full-thickness small intestinal biopsy specimens with a 45-mm ELS may be an effective and safe technique for use in healthy adult experimental horses.

Objective: To evaluate the effects of endotoxin administered IV on glucose and insulin dynamics in horses., Animals: 16 healthy adult mares., Procedures: Each week of a 2-week randomized crossover study, each horse received an IV injection (duration, 30 minutes) of Escherichia coli O55:B5 lipopolysaccharide (LPS) in 60 mL of sterile saline (0.9% NaCl) solution (20 ng/kg) or sterile saline solution alone (control treatment). Frequently sampled IV glucose tolerance test procedures were performed at 24 hours before (baseline) and 24 and 48 hours after injection; glucose and insulin dynamics were assessed via minimal model analysis., Results: 13 of 16 horses had a clinical response to LPS, which was characterized by mild colic and leukopenia. Before treatment, mean +/- SD insulin sensitivity was 2.9 +/- 1.9 x 10(4) L x min(1) x mU(1); this significantly decreased to 0.9 +/- 0.9 x 10(4) L x min(1) x mU(1) 24 hours after treatment (69% reduction) and was 1.5 +/- 0.9 x 10(4) L x min(1) x mU(1) 48 hours after treatment. At baseline, mean +/- SD acute insulin response to glucose was 520 +/- 196 mU x min x L(1); this significantly increased to 938 +/- 620 mU x min x L(1) (80% increase) and 755 +/- 400 mU x min x L(1) (45% increase) at 24 and 48 hours after LPS treatment, respectively., Conclusions and Clinical Relevance: Compared with baseline values, insulin sensitivity was decreased for 24 hours after IV injection of LPS, and affected horses had a compensatory pancreatic response. These disturbances in glucose and insulin dynamics may contribute to development of laminitis in horses.

Objective: To determine the effects of dexamethasone treatment on selected components of insulin signaling and glucose metabolism in skeletal muscle obtained from horses before and after administration of a euglycemic-hyperinsulinemic clamp (EHC)., Animals: 6 adult Standardbreds., Procedures: In a balanced crossover study, horses received either dexamethasone (0.08 mg/kg, IV, q 48 h) or an equivalent volume of saline (0.9% NaCl) solution, IV, for 21 days. A 2-hour EHC was administered for measurement of insulin sensitivity 1 day after treatment. Muscle biopsy specimens obtained before and after the EHC were analyzed for glucose transporter 4, protein kinase B (PKB), glycogen synthase kinase (GSK)-3alpha/beta protein abundance and phosphorylation state (PKB Ser(473) and GSK-3alpha/beta Ser(21/9)), glycogen synthase and hexokinase enzyme activities, and muscle glycogen concentration., Results: Dexamethasone treatment resulted in resting hyperinsulinemia and a significant decrease (70%) in glucose infusion rate during the EHC. In the dexamethasone group, increased hexokinase activity, abrogation of the insulin-stimulated increase in glycogen synthase fractional velocity, and decreased phosphorylation of GSK-3alpha Ser(21) and GSK-3B Ser(9) were detected, but there was no effect of dexamethasone treatment on glucose transporter 4 content and glycogen concentration or on PKB abundance and phosphorylation state., Conclusions and Clinical Relevance: In horses, 21 days of dexamethasone treatment resulted in substantial insulin resistance and impaired GSK-3 phosphorylation in skeletal muscle, which may have contributed to the decreased glycogen synthase activity seen after insulin stimulation.

Objective: To determine effects of dexamethasone on glucose dynamics and insulin sensitivity in healthy horses., Animals: 6 adult Standardbreds., Procedures: In a balanced crossover study, horses received dexamethasone (0.08 mg/ kg, IV, q 48 h) or an equivalent volume of saline (0.9% NaCl) solution (control treatment) during a 21-day period. Horses underwent a 3-hour frequently sampled IV glucose tolerance test (FSIGT) 2 days after treatment. Minimal model analysis of glucose and insulin data from FSIGTs were used to estimate insulin sensitivity (Si), glucose effectiveness (Sg), acute insulin response to glucose (AIRg), and disposition index. Proxies for Si (reciprocal of the inverse square of basal insulin concentration [RISQI]) and beta-cell responsiveness (modified insulin-to-glucose ratio [MIRG]) were calculated from basal plasma glucose and serum insulin concentrations., Results: Mean serum insulin concentration was significantly higher in dexamethasone-treated horses than control horses on days 7, 14, and 21. Similarly, mean plasma glucose concentration was higher in dexamethasone-treated horses on days 7, 14, and 21; this value differed significantly on day 14 but not on days 7 or 21. Minimal model analysis of FSIGT data revealed a significant decrease in Si and a significant increase in AIRg after dexamethasone treatment, with no change in Sg or disposition index. Mean RISQI was significantly lower, whereas MIRG was higher, in dexamethasone-treated horses than control horses on days 7, 14, and 21., Conclusions and Clinical Relevance: The study revealed marked insulin resistance in healthy horses after 21 days of dexamethasone administration. Because insulin resistance has been associated with a predisposition to laminitis, a glucocorticoid-induced decrease in insulin sensitivity may increase risk for development of laminitis in some horses and ponies.

Objective: To compare effects of low and high intensity warm-up exercise on oxygen consumption (VO2) and carbon dioxide production (VCO2) in horses., Animals: 6 moderately conditioned adult Standard-breds., Procedures: Horses ran for 2 minutes at 115% of maximum oxygen consumption (VO2max), 5 minutes after each of the following periods: no warm-up (NoWU); 10 minutes at 50% of VO2max (LoWU); or 7 minutes at 50% VO2max followed by 45-second intervals at 80, 90, and 100% VO2max (HiWU). Oxygen consumption and VCO2 were measured during exercise, and kinetics of VO2 and VCO2 were calculated. Accumulated O2 deficit was also calculated., Results: For both warm-up trials, the time constant for the rapid exponential increase in VO2 was 30% lower than for NoWU. Similarly, the rate of increase in VCO2 was 23% faster in LoWU and HiWU than in NoWU. Peak values for VO2 achieved during the high-speed test were not significantly different among trials (LoWU, 150.2 +/- 3.2 ml/kg/min; HiWU, 151.2 +/- 4.2 ml/kg/min; NoWU, 145.1 +/- 4.1 ml/kg/min). However, accumulated O2 deficit (ml of O2 equivalents/kg) was significantly lower during LoWU (65.3 +/- 5.1) and HiWU (63.4 +/- 3.9) than during NoWU (82.1 +/- 7.3)., Conclusions and Clinical Relevance: Both the low- and high-intensity warm-up, completed 5 minutes before the start of high-intensity exercise, accelerated the kinetics of VO2 and VCO2 and decreased accumulated O2 deficit during 2 minutes of intense exertion in horses that were moderately conditioned.

Objective: To quantify total fluid loss in sweat of Thoroughbreds during >3 hours of low-intensity exercise in controlled conditions and to calculate and compare estimated ion losses in sweat, according to 3 methods., Animals: 6 exercise-trained Thoroughbreds., Procedure: Fluid and ion losses in sweat were measured in 6 horses exercising at 40% of the speed that elicited maximum oxygen consumption for 45 km. Horses were given a 15-minute rest period at the end of three 15-km exercise phases. Horses completed 2 exercise trials. Ion losses in sweat were calculated, using measurements of local sweating rate and sweat ion composition (SWT), change in net exchangeable cation content (CAT), and change in extracellular ion content (PLAS) derived from plasma total solids and ion concentrations., Results: Measurement of SWT revealed a mean (+/- SEM) fluid loss in sweat during 45 km of exercise of 27.5 +/- 1.6 L. Total ion loss in sweat was approximately 241 g or 7.8 mol with higher sodium losses in the second and third phases of exercise compared with the first phase. Losses of sodium and potassium calculated by SWT or CAT were not significantly different from each other, whereas losses of these ions as determined by PLAS were significantly lower., Conclusions and Clinical Relevance: Calculation of ion losses from a mean whole body sweating rate extrapolated from either local sweating rate and sweat ion composition or from change in net exchangeable cation content provide similar results, whereas ion losses determined by changes in extracellular ion content derived from plasma total solids and ion concentration results in underestimation of actual losses.

Objective: To determine whether supplemental i.v. calcium administration would attenuate or prevent gentamicin-induced acute renal failure, defined as an increase in serum creatinine concentration > or = 50% above baseline., Animals: 10 healthy pony mares., Procedure: Pony mares were randomly assigned to receive calcium at a dosage of 20 mg/kg of body weight or saline solution i.v., twice daily for 14 days. All pony mares received gentamicin at a dosage of 20 mg/kg i.v. every 8 hours for 14 days. Gentamicin pharmacokinetic, serum biochemical, and urinalysis data were measured every other day for the 14-day study period. Renal histologic examination was performed, and results were scored at the end of the 14-day period., Results: 4 of 5 mares not receiving calcium supplementation developed acute renal failure. Only 1 of the 5 mares receiving calcium supplementation developed acute renal failure. Over the course of the study, pony mares receiving calcium supplementation had significantly fewer changes in urinalysis variables, and significantly less microscopic renal damage., Conclusion: Daily i.v. administration of calcium attenuated gentamicin-induced acute renal failure., Clinical Relevance: Calcium supplementation may help diminish the risk of acute renal failure associated with aminoglycoside antibiotics.

Objective: To compare dew-point hygrometry, direct sweat collection, and measurement of body water loss as methods for determination of sweating rate (SR) in exercising horses., Animals: 6 exercise-trained Thoroughbreds., Procedure: SR was measured in 6 horses exercising at 40% of the speed that elicited maximum oxygen consumption for 45 km, with a 15-minute rest at the end of each 15-km phase. Each horse completed 2 exercise trials. Dew-point hygrometry, as a method of local SR determination, was validated in vitro by measurement of rate of evaporative water loss. During exercise, local SR was determined every 10 minutes by the following 2 methods: (1) dew-point hygrometry on the neck and lateral area of the thorax, and (2) on the basis of the volume of sweat collected from a sealed plastic pouch attached to the lateral area of the thorax. Mean whole body SR was calculated from total body water loss incurred during exercise., Results: Evaporation rate measured by use of dew-point hygrometry was significantly correlated (r2 = 0.92) with the actual rate of evaporative water loss. There was a similar pattern of change in SR measured by dew-point hygrometry on the neck and lateral area of the thorax during exercise, with a significantly higher SR on the neck. The SR measured on the thorax by direct sweat collection and by dew-point hygrometry were of similar magnitude. Mean whole body SR calculated from total body water loss was not significantly different from mean whole body SR estimated from direct sweat collection or dew-point hygrometry measurements on the thorax., Conclusions: Dew-point hygrometry and direct sweat collection are useful methods for determination of local SR in horses during prolonged, steady-state exercise in moderate ambient conditions. Both methods of local SR determination provide an accurate estimated of whole body SR.

Objectives: To determine whether pentoxifylline treatment altered hematologic, rheologic, electrolyte, or blood gas test results of Thoroughbreds during submaximal treadmill exercise., Animals: 5 healthy Thoroughbreds that had raced within the past year and had no history of exercise-induced pulmonary hemorrhage., Procedure: Mixed venous blood samples were obtained before exercise, at treadmill speeds of 9 and 13 m/s, and 20 minutes after exercise; hematologic, rheologic, electrolyte, and blood gas test results were determined., Results: Pentoxifylline treatment resulted in a 45% reduction in RBC filtration pressures for horses at rest. The improved RBC filterability persisted during treadmill exercise. Horses treated with pentoxifylline had a greater decrease in Po2 values and a lesser increase in plasma lactate concentration during treadmill exercise., Conclusion: Administration of pentoxifylline improved RBC deformability of horses at rest and during treadmill exercise., Clinical Relevance: Improved RBC deformability resulting from pentoxifylline treatment may reduce exercise-associated shear stress in pulmonary capillaries, thereby attenuating exercise-induced pulmonary hemorrhage.

Objective: To determine whether furosemide treatment altered the blood flow properties and serum and RBC electrolyte concentrations of Thoroughbreds during submaximal treadmill exercise., Design: Thoroughbreds were subjected to submaximal treadmill exercise with and without treatment with furosemide (1 mg/kg of body weight, IV)., Animals: 5 healthy Throughbreds that had raced within the past year and had no history of exercise-induced pulmonary hemorrhage., Procedure: Venous blood samples were obtained before exercise, at treadmill speeds of 9 and 13 m/s, and 10 minutes after exercise, and hemorheologic and electrolyte test results were determined., Results: Hemorheologic changes 60 minutes after furosemide administration included increased PCV, plasma total protein concentration, whole blood viscosity, mean RBC volume, and RBC potassium concentration, and decreased serum potassium concentration, serum chloride concentration, and RBC chloride concentration. Furosemide treatment attenuated the exercise-associated changes in RBC size, serum sodium concentration, serum potassium concentration, RBC potassium and chloride concentrations, and RBC density; exacerbated exercise-associated increases in whole blood viscosity; and had no effect on RBC filterability., Conclusions: The hemorheologic effects of furosemide probably occurred secondary to total body and transmembrane fluid and electrolyte fluxes and would not improve blood flow properties., Clinical Relevance: The beneficial effects of furosemide treatment in reducing the severity of bleeding in horses with exercise-induced pulmonary hemorrhage cannot be explained by improved blood flow properties.

Hemorheologic alterations induced by incremental treadmill exercise were examined in 5 Thoroughbreds. Blood viscosity; PCV; RBC filterability, density gradient profile, and shape; serum and RBC electrolyte concentrations; and plasma total solids and lactate concentrations were measured before exercise, at treadmill speeds of 9 and 13 m/s, and 10 minutes after exercise. Exercise was associated with significant (P < 0.05) increases in PCV, blood viscosity, and plasma total solids concentration. After adjustment of PCV to 40% by adding or removing each horse's own plasma, blood viscosity remained significantly greater in the sample obtained at 13 m/s, compared with that in samples taken at rest. Filterability of RBC was significantly decreased at 13 m/s, compared with values from other sampling times. During exercise, a significantly greater proportion of the RBC were less dense and were found in the upper layers of the RBC density gradient profile, compared with resting values. This change was associated with a significant increase in RBC mean cell volume. Rapid increases in serum sodium and potassium concentrations during exercise were accompanied by significant increases in RBC potassium and chloride concentrations. This study revealed a consistent pattern of hemorheologic alterations associated with exercise in Thoroughbreds, suggesting that multiple hemorheologic tests are needed to adequately define these complex alterations during exercise in horses.

The hypothesis that equine laminitis is caused by thrombosis of vessels in the laminar corium (dermis) was investigated. Hemostatic alterations were evaluated by determining platelet count, platelet survival, platelet adhesiveness to vascular subendothelium, activated clotting time, and whole blood recalcification time. Thrombosis of vessels in the hoof wall was evaluated by scintigraphic studies of the hoof wall after administration of indium-111 (111In)-labeled platelets, contrast arteriography, and histologic examination. Platelet count remained constant before and at the onset of lameness; however, survival of 111In-labeled platelets was shortened. Scintigraphy of affected feet revealed accumulation of 111In-labeled platelets distal to the coronary band. Arteriography of disarticulated saline-perfused feet revealed marked reduction in blood supply to affected hooves. Histologic examination of the laminar dermis disclosed variable numbers of microthrombi in dermal veins of affected feet from 3 of 4 ponies with laminitis. Whole blood recalcification time was shortened at 8 hours after administration of carbohydrate and was prolonged at the onset of laminitis. Activated clotting time was prolonged at 32 hours after carbohydrate administration and at the onset of lameness. Plasma endotoxin-like activity was detected in 1 of 4 affected ponies. These data confirm that microvascular thrombosis existed at the onset of lameness in ponies with carbohydrate-induced laminitis and indicate that systemic coagulopathy may have preceded development of thrombosis.

Effects of echinocytosis on blood rheology and exercise performance were evaluated for 5 Thoroughbreds. Echinocytosis was induced by administration of furosemide (1 mg/kg of body weight, IM, q 12 h) for 4 days. Furosemide treatment resulted in decreases in serum sodium and serum chloride concentrations and in RBC chloride and potassium concentrations. Echinocytosis was associated with increased RBC density as determined by RBC density gradient centrifugation. However, samples containing echinocytes were more filterable than control samples, indicating that echinocytes were not rigid cells. Erythrocyte sedimentation rate was decreased in blood samples containing echinocytes, indicating that cell-to-cell interaction was reduced. Whole blood viscosity was not altered by presence of echinocytes. Echinocytes did not impair the capacity of horses to complete treadmill exercise tests, nor did they alter heart rate or blood gas variables. However, plasma lactate concentration was higher in samples obtained during exercise at a treadmill speed of 9 m/s. Echinocytosis was associated with higher postrace creatine kinase activity. These data indicate that echinocytes may be dense, but not rigid cells, which have decreased tendency to aggregate and do not increase whole blood viscosity. Therefore, echinocytes are unlikely to inhibit or obstruct microvascular blood flow.

A commercially available radioimmunoassay kit for measurement of human osteocalcin was validated for use in horses. For accurate measurement of equine serum osteocalcin, blood samples may be collected at a temperature between 20 and 25 C, then centrifuged within 90 minutes; serum may be stored at -20 C in plastic tubes for up to 26 weeks. Serum may be thawed and refrozen up to 5 times without significant change in measured equine serum osteocalcin concentration. Assay sensitivity was 0.16 ng/ml. Recovery of bovine osteocalcin standard added to equine serum was linear. Intra-assay coefficient of variation (x 100) for 2 equine serum pools was 6.9 (mean +/- SD, 13.9 +/- 1.0 ng/ml) and 7.5 (10.6 +/- 0.8 ng/ml) %. Interassay coefficient of variation for 3 equine serum pools measured in 12 assays was 12.5 (16.1 +/- 2.0 ng/ml), 12.7 (11.5 +/- 1.5 ng/ml), and 24.6 (3.0 +/- 0.7 ng/ml) %. Dilutional parallelism was documented by assaying pooled equine serum at 4 dilutions and correcting the mean result for dilution. Significant change was not observed in equine serum osteocalcin concentration for various time-of-day blood sample collections in horses housed under continuous lighting.

The effects of furosemide and pentoxifylline on blood flow properties in horses were investigated. Hematologic and rheologic changes were examined in 4 horses before and 3 minutes after administration of epinephrine (1 mg, IV). The next day, hemorheologic changes were determined before and 3 hours after administration of furosemide (1 mg/kg of body weight, IM), and after administration of epinephrine at the sampling at 3 hours. Hematologic and rheologic changes were evaluated weekly in 3 horses given pentoxifylline (8.5 mg/kg, q 12 h, PO) for 28 days. In addition, hemorheologic responses to epinephrine were determined on days 0, 14, and 28 of pentoxifylline treatment. Neutrophil filtration studies were also performed 2 hours after IV administration of pentoxifylline (8.5 mg/kg). Postepinephrine values for PCV, RBC and WBC counts, and blood viscosity were greater than preepinephrine values. Erythrocyte sedimentation rates decreased after epinephrine, whereas RBC filterability did not change. Treatment with furosemide was associated with increases in mean RBC hemoglobin concentration and blood viscosity. Filterability of RBC did not change. Treatment with pentoxifyllie resulted in an increase in RBC filterability and erythrocyte sedimentation rate and a decrease in PCV; however, mean values for hematocrit and RBC count did not change. Treatment with pentoxifylline did not result in a change in resting blood viscosity, but markedly reduced the postepinephrine increase in blood viscosity. Neither IV nor orally administered pentoxifylline had an effect on neutrophil filtration. It was concluded that pentoxifylline has beneficial effects on RBC filterability and postepinephrine changes in blood viscosity, which may contribute to improvements of microcirculatory blood flow. In addition, furosemide may exacerbate exercise-associated hyperviscosity in horses.