Dietary-induced metabolic acidosis decreases bone mineral density in mature ovariectomized ewes.

Dietary-induced metabolic acidosis (DIMA) may be a significant confounder in the development of osteoporosis. Diets that are acidifying are typically rich in proteins and grains and relatively poor in fruits and vegetables. Previous studies have not examined whether an interaction between estrogen depletion and DIMA have a compounded affect on bone mineral density loss. Sheep have been used successfully in previous studies to examine the interaction of bone turnover and ovariectomy. Therefore, the goal of this pilot study was to determine if bone mineral density (BMD) loss could be induced using DIMA in skeletally mature ovariectomized (OVX) ewes.

Assessment of subchondral bone mineral density in equine metacarpophalangeal and stifle joints.

Functional relationships between articular cartilage and underlying subchondral bone have been shown to be associated with the progression of osteoarthritis (OA). However, quantifiable factors substantiating this relationship do not exist. Therefore, the study objective involved quantifying subchondral bone mineral density (BMD) of the equine metacarpophalangeal (MCP) and stifle joints as a step in determining if regional subchondral BMD may be associated with OA changes. BMD was bilaterally quantified using dual energy xray absorptiometry (DEXA) for four subchondral regions: palmer and dorsal aspect of the medial 3rd metacarpal (MC3P, MC3D), medial femoral condyle (MFC), and the medial trochlear ridge (MTR). BMD (g/cm2) was measured using a 5.2 x 5.2 mm region of interest in the subchondral bone area. To account for the differences in thickness, BMD (g/cm2) was divided by sample thickness providing volumetric BMD (g/cm3). No significant effect of side on BMD values was found (p > 0.72). However, there were significant differences in BMD found between all regions (p < 0.05). The volumetric BMD of the MC3P region was significantly greater than the MC3D and the MC3D was greater than the MFC, while the MFC was greater than the MTR (MC3P > MC3D > MFC > MTR). The MCP regions had a higher volumetric BMD than the stifle regions possibly due to higher weight distribution, smaller articular surface area, and joint geometry. The adaptive bone structural variations between the regions studied were shown to be significant. Mechanical properties of the overlying cartilage are currently being analyzed to correlate with these results and provide a possible diagnostic method to investigate OA progression.

Central and peripheral temperature changes in sheep following ovariectomy.

OBJECTIVES: To determine if ovariectomized ewes undergo periodic body temperature rises (hot flashes) similar to women at menopause. METHODS: Eighteen mature ewes were assigned to ovariectomy (OVX), ovariectomy+17 beta-estradiol implant (OVXE) or Sham. Electronic temperature loggers placed subcutaneously over the carotid artery and within the abdomen (core) and subcutaneously in the thigh and axilla (peripheral) were programmed to record body temperatures every 2.5 min for 59 days. Circadian rhythm changes were avoided by dividing readings into 1 h intervals. Hot flashes were defined as a 0.2 or 0.4 degrees C increase over the minimum temperature recorded for a 1 h interval for each sheep. RESULTS: Logger placement did not reflect core and peripheral temperatures. The carotid and abdominal sites were most useful. The percentage of readings considered HF at the carotid site was 63% OVX, 54% OVXE and 37% Sham (P<0.001), and at the abdominal site were 32% OVX, 15% OVE and 17% Sham (P<0.001). When only the first 7 days after ovariectomy were analyzed, the percentage of readings considered to be HF at the carotid site was 75% OVX, 63% OVXE, and 49% Sham (P<0.001), and at the abdominal site was 35% OVX, 15% OVXE and 17% Sham (P<0.001). CONCLUSIONS: Ovariectomy in the ewe does illicit changes in body temperature compared with control ewes, which may be interpreted as HF. However, shifts in the circadian rhythm were not apparent. Estradiol treatment led to milder and less frequent HF. Periodic HF in species other than rats have heretofore not been reported.

Neonatal isoerythrolysis involving the Qc and Db antigens in a foal.

In 1992, a multiparous 13-year-old Thoroughbred mare and her 48-hour-old colt were examined because of possible neonatal isoerythrolysis (NI). Supportive treatment was administered, and the foal recovered without requiring a transfusion. According to the owners, the mare had delivered foals without incident during 1987 and 1991. The mare was barren during 1993, but in 1994, delivered a filly that developed severe NI. The foal was given 3 transfusions and eventually recovered without complications. Blood typing analysis of the mare and its foals indicated that all 4 foals were positive for the Qc, Db, and Dq antigens, and the 3 most recently born foals were positive for the Ua antigen; however, the marc was negative for the Qc, Db, Dq, and Ua antigens. The mare did not have alloantibodies against Ua and did not react to the Dq antigen. However, in 1994, the mare reacted against the Db (the reaction was characterized by strong agglutination and an increase in titer at the time of parturition and a subsequent decrease) and Qc (the reaction was characterized by weak lysis and an increase in titer at the time of parturition and a subsequent decrease) antigens. Results of testing in this mare and foals suggested that although approximately 90% of all cases of NI in horses are attributable to the Aa or Qa antigen, other antigens may be involved.

Effect of ration and exercise on plasma creatine kinase activity and lactate concentration in Thoroughbred horses with recurrent exertional rhabdomyolysis.

OBJECTIVE: To determine the effects of 3 rations (low grain, fat, high grain) on plasma creatine kinase (CK) activity and lactate concentration in Thoroughbred horses with recurrent exertional rhabdomyolysis (RER). ANIMALS: 5 Thoroughbreds with RER and 3 healthy Thoroughbreds (control horses). PROCEDURES: Rations were formulated to meet (low-grain and fat rations) or exceed (high-grain ration) daily energy requirements. Each ration was fed to horses in a crossover design for 3 weeks. Horses were exercised on a treadmill Monday through Friday; maximum speed on Monday and Friday was 11 m/s (6% slope), on Tuesday and Thursday was 9 m/s, and on Wednesday was 4.5 m/s. Plasma CK activity and lactate concentration were determined before and after exercise. RESULTS: Horses with RER fed the high-grain ration had significantly greater CK activity and change in CK activity 4 hours after exercise, compared with those fed the low-grain ration. Horses with RER exercised at the trot or canter had significantly greater increases in CK activity, compared with those exercised at the gallop. Plasma lactate concentrations after exercise were similar in control and affected horses. Lactate concentration and CK activity were not correlated in horses with RER. CONCLUSIONS AND CLINICAL RELEVANCE: Rations high in grain and formulated to exceed daily energy requirements may increase episodes of rhabdomyolysis in thoroughbred horses susceptible to RER.

Glucose uptake in horses with polysaccharide storage myopathy.

OBJECTIVE: To determine whether excessive glycogen accumulation in skeletal muscle of Quarter Horses with polysaccharide storage myopathy (PSSM) is a result of enhanced cellular uptake of glucose. ANIMALS: 6 horses with PSSM and 10 healthy (control) horses. PROCEDURE: Intravenous glucose tolerance tests (IVGTT), oral glucose tolerance tests (OGTT), and modified insulin tolerance tests (MITT) were performed. Plasma glucose and insulin concentrations were measured in blood samples collected before and for up to 8 hours after glucose or insulin administration. RESULTS: Peak glucose concentrations during IVGTT were similar for both groups of horses, but rate of glucose clearance was 1.5 times faster in horses with PSSM than in controls. Moreover, circulating concentrations of insulin before and after glucose injection were lower in the PSSM group. Blood glucose concentrations from minute 90 to minute 300 of the OGTT were lower in horses with PSSM than in controls. The MITT resulted in acute decreases in blood glucose concentrations in both groups of horses; however, horses with PSSM sustained low blood glucose concentrations for more than 3 hours after insulin injection, whereas blood glucose concentrations in controls returned to baseline values within 2 hours. CONCLUSIONS: Quarter Horses with PSSM have enhanced cellular uptake of glucose that may be, in part, caused by an increased sensitivity to insulin. CLINICAL RELEVANCE: Horses with PSSM have an increased rate of glucose clearance in response to insulin secretion. Thus, diets low in soluble carbohydrate may be the most effective way to decrease glycogen accumulation in skeletal muscle of these horses.

Heritability of recurrent exertional rhabdomyolysis in Thoroughbred racehorses.

OBJECTIVE: To determine the likely mode of inheritance and identify probable foundation horses for recurrent exertional rhabdomyolysis (RER) in Thoroughbred (TB) racehorses. ANIMALS: 4 families of TB racehorses with a high prevalence of RER, consisting of 3 to 53 horses/family, were used to determine mode of inheritance. Sixty-two TB horses with RER and 34 control TB racehorses without RER were used to identify probable foundation horses for the RER trait. PROCEDURE: RER was diagnosed by a veterinarian and verified by detecting high serum creatine kinase activity. Pedigrees dating from 1930 for all horses were entered into a database. Pedigrees of horses in 4 families were visually inspected for a pattern of inheritance and used for calculation of foundation horse contributions and inbreeding coefficients. The Markov chain Monte Carlo technique was used to analyze pedigrees of the 62 affected and 34 control horses for the conditional probability of foundation genotypes. A dominant mode of inheritance with variable expression model was used. RESULTS: Pedigree analysis supported an autosomal dominant mode of inheritance with variable expression. All affected horses from the 4 families shared a common ancestor. This ancestor and 5 other stallions had a conditional probability of 1.00 for being affected. All 6 stallions shared a common male ancestor within 3 to 5 generations. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of this study, the RER trait has been in TB racehorses for more than 70 years and may be inherited as an autosomal dominant trait with variable expression.

Skeletal muscle metabolic response to exercise in horses with 'tying-up' due to polysaccharide storage myopathy.

Polysaccharide storage myopathy (PSSM) is a distinct cause of exertional rhabdomyolysis in Quarter Horses that results in glycogen and abnormal polysaccharide accumulation. The purpose of this study was to determine if excessive glycogen storage in PSSM is due to a glycolytic defect that impairs utilisation of this substrate during exercise. Muscle biopsies, blood lactates and serum CK were obtained 1) at rest from 5 PSSM Quarter Horses, 4 normal Quarter Horses (QH controls) and 6 Thoroughbreds with recurrent exertional rhabdomyolysis (TB RER) and 2) after a maximal treadmill exercise test in PSSM and QH controls. In addition, 3 PSSM horses performed a submaximal exercise test. At rest, muscle glycogen concentrations were 2.4x and 1.9x higher in PSSM vs. QH controls or TB RER, respectively. Muscle lactates at rest were similar between PSSM and QH controls but significantly higher in PSSM vs. TB RER. Muscle glucose-6-phosphate concentrations were also higher in PSSM horses than controls combined. During maximal exercise, mean muscle glycogen concentrations declined 2.7x more and mean lactate increased 2x more in PSSM vs. QH controls; however, differences were not statistically significant. Blood lactate concentrations after maximal exercise did not reflect generally higher muscle lactate in PSSM vs. QH controls. No change in blood lactate concentrations occurred in PSSM horses with submaximal exercise. Serum CK activity increased significantly 4 h after maximal and submaximal exercise and was significantly higher in PSSM vs. QH controls. These results show that during maximal exercise, PSSM horses utilised muscle glycogen and produce lactic acid via a functional glycolytic pathway and that during submaximal exercise oxidative metabolism was unimpaired. The excessive glycogen storage and formation of abnormal polysaccharide in PSSM horses therefore appear to reflect increased glycogen synthesis rather than decreased utilisation. The specific subset of horses with exertional rhabdomyolysis due to PSSM would likely benefit clinically from a diet low in soluble carbohydrates like grain with fat added as well as gradually increasing daily exercise to reduce excessive glycogen accumulation and enhance utilisation.

Epidemiologic analysis of factors influencing exertional rhabdomyolysis in Thoroughbreds.

OBJECTIVE: To determine incidence, effect on performance, and management practices associated with exertional rhabdomyolysis (ER) in Thoroughbreds. SAMPLE POPULATION: Medical records for 984 Thoroughbreds and a survey of trainers of horses with and without ER. PROCEDURES: Medical records for 984 Thoroughbreds stabled at a midwestern racetrack were examined to determine the incidence of ER during the 1995 racing season. A retrospective questionnaire was administered to trainers to determine management practices associated with ER in 59 Thoroughbreds with ER and 47 control Thoroughbreds in training. Multiple logistic regression was used to determine management factors associated with ER. RESULTS: ER affected 48 of 984 (4.9%) Thoroughbreds. Two-year-old females were most frequently affected, and 36 of 96 (37.5%) trainers had > or = 1 horse with ER. Horses with ER were more likely not to race during the racing season, compared with control horses. For horses that raced, differences were not found with respect to racing performance between ER and control horses. Exertional rhabdomyolysis developed frequently in susceptible horses that had > or = 1 day of rest prior to exercise and that galloped during exercise. Horses with ER were commonly fed > 4.5 kg of grain daily. Nervous and extremely nervous horses were 5.4 times more likely, and horses with some form of lameness were 4.2 times more likely, to have ER. CONCLUSIONS AND CLINICAL RELEVANCE: Exertional rhabdomyolysis is common in Thoroughbreds, and ER can be affected by temperament, sex, age, diet, exercise routines, and lameness. Management that minimizes excitability, particularly when tailored to each horse, may be most effective for controlling ER.

Effect of diet on thoroughbred horses with recurrent exertional rhabdomyolysis performing a standardised exercise test.

Previous studies have associated recurrent exertional rhabdomyolysis (RER) with a diet high in soluble carbohydrate (CHO). The purpose of this study was to investigate the effect of 3 diets on clinical and metabolic parameters in 5 Thoroughbred horses with RER and 3 healthy Thoroughbreds performing a standardised exercise test (SET). Two diets were formulated to meet energy requirements for the amount of exercise being performed in the form of CHO or fat (21.4 Mcal DE/day). The third diet was formulated to provide 135% of the DE of the other 2 diets in the form of an excessive amount of carbohydrate (28.8 Mcal DE/day). Diets were fed in a crossover design for 3 week blocks and then horses performed a near maximal SET. Changes in heart rate (HR), plasma lactate, plasma glucose, total plasma solids, packed cell volume (PCV), muscle lactate and muscle glycogen concentration were measured immediately prior to, during, and 5 min after exercise. Serum creatine kinase (CK) activity was measured prior to and 4 h post SET. A 2-way ANOVA was used to examine the effect of group and dietary treatment. When dietary treatments were compared, horses fed the high-CHO diet had a mean pre-SET PCV and pre-SET HR that was higher than horses fed the fat diet (P = 0.06 and P = 0.07, respectively). Pre-SET heart rates were highest in RER horses consuming the high-CHO diet compared to RER horses consuming the low-CHO and fat diets (P = 0.02). Horses with RER had 4 h post SET CK activity greater than 400 u/l in 7/14 (50%) measurements compared to control horses which had CK activity greater than 400 u/l in 2/7 (29%) measurements. This study did not demonstrate a significant effect of diet on rhabdomyolysis, indicated by CK activity, or on the metabolic response to exercise. However, diet may have a calming effect on Thoroughbred horses with RER as manifested by decreased pre-exercise heart rates and decreased pre-exercise PCV in horses fed the fat diet.

Exertional rhabdomyolysis in quarter horses and thoroughbreds: one syndrome, multiple aetiologies.

The purpose of this study was to determine if chronic exertional rhabdomyolysis (ER) in Quarter Horses and Thoroughbreds represents one or several distinct myopathies. Eighteen Quarter Horses and 18 Thoroughbreds with ER were selected from cases presented to the Veterinary Hospital on the basis of a history of ER, assessment of muscle histopathology, and serum CK activity before and 4 h post exercise. In addition, 2 of 3 of the following parameters were evaluated: muscle glycogen concentrations, thyroid hormones (T3, T4), fractional excretion (FE) of sodium, potassium and chloride. The CK response to training, the metabolic response to a near maximal standardised exercise test (SET), blood glucose concentrations after an i.v. glucose challenge and a skeletal muscle in vitro caffeine contracture test were performed on 5 of the Quarter Horses, selected because of polysaccharide storage myopathy (PSSM), and 5 of the Thoroughbreds. Serum T3 and T4 were all within normal limits. Low FE of sodium and potassium were seen in < 20% of Quarter Horses and Thoroughbreds. Four hours post exercise, CK was increased in 77% of Quarter Horses and 72% of Thoroughbreds with ER. Muscle glycogen concentrations in Quarter Horses with ER were significantly higher than in normal Quarter Horses and Thoroughbreds with ER. No Thoroughbreds, but 15/18 Quarter Horses with ER had abnormal polysaccharide accumulation in muscle biopsies consistent with a diagnosis of PSSM. PSSM Quarter Horses had higher CK activity during training than Thoroughbreds and higher glycogen utilisation with the SET. PSSM Quarter Horses also had significantly enhanced glucose clearance compared to normal Quarter Horses and Thoroughbreds with ER. Thoroughbreds with ER had significantly lower thresholds for caffeine-induced contracture than normal horses and PSSM Quarter Horses. It was concluded that there are multiple causes for exertional rhabdomyolysis. In Quarter Horses, rhabdomyolysis is commonly due to a glycogen storage disorder, PSSM, and is readily expressed in untrained horses. In Thoroughbreds, ER is commonly due to an underlying abnormality of muscle contraction. Rhabdomyolysis in Thoroughbreds, however, is only expressed intermittently when key stressors are present.

Transient alteration in intestinal permeability to technetium Tc99m diethylenetriaminopentaacetate during the prodromal stages of alimentary laminitis in ponies.

OBJECTIVES: To determine whether mucosal permeability is altered during the prodromal stages of alimentary laminitis. ANIMALS: 15 healthy adult ponies. PROCEDURES: intestinal permeability was evaluated for control ponies (n = 5) and for ponies 4 to 12 (n = 5) and 20 to 28 (n = 5) hours after administration of carbohydrate overload. Mucosal permeability was determined by measuring the percentage of orally administered technetium Tc99m diethylenetriaminopentaacetate (99mTc-DTPA) excreted in urine during an 8-hour period, then measuring blood radioactivity at hourly intervals. Plasma endotoxin-like activity was measured by use of a chromogenic Limulus amebocyte assay. RESULTS: Urinary excretion of 99mTc-DTPA was 2.45% of administered dose for control ponies, and was 16.67% of administered dose 4 to 12 hours and 3.57% of administered dose 20 to 28 hours after administration of carbohydrate. CONCLUSIONS: A marked but transient increase in intestinal permeability was observed early in the prodromal stages of alimentary laminitis. CLINICAL RELEVANCE: Absorption of substances from the intestine may be an initiating event in alimentary laminitis.

Type-II renal tubular acidosis and ventricular tachycardia in a horse.

A 14-year-old Arabian mare was admitted for lethargy, anorexia, and low fecal output. On the basis of laboratory, physical examination, and electrocardiographic findings, diagnoses of type-II renal tubular acidosis (RTA), impaction of the large colon, and ventricular tachycardia were made. Diagnosis of type-II RTA was based on measurement of a low fractional excretion value for potassium and fractional excretion value for sodium within the reference range. In contrast, horses with type-I RTA have high fractional excretion values for sodium and fractional excretion values for potassium within reference ranges. Treatment consisted of intravenous and oral administration of sodium bicarbonate, intravenous administration of fluids, and oral administration of mineral oil and docusate sodium. Acidosis improved, and ventricular tachycardia resolved with resolution of acidosis. Oral administration of bicarbonate was continued after discharge. The mare had several relapses, which were associated with anorexia and low intake of supplemental bicarbonate. The mare was found dead 2 months after discharge.

Results of quantitative cultures of urine by free catch and catheterization from healthy adult horses.

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

Acquired B lymphocyte deficiency and chronic enterocolitis in a 3-year-old quarter horse.

This case report describes a 3-year-old American Quarter Horse with acquired immunodeficiency. Clinical signs included chronic diarrhea due to Salmonella typhimurium and bacterial pneumonia. Characterization of the immunodeficiency involved in vivo phytohemagglutinin (PHA) intradermal testing, in vitro lymphocyte proliferation in response to concanavalin A, immunofluorescence flow cytometry data on blood lymphocytes, serum protein electrophoresis and immunoglobulin (Ig) quantification. A diagnosis of B lymphocyte deficiency with resulting deficiencies in serum IgG, IgA and IgM and a concurrent decrease in T cell function was made based on these tests. Postmortem examination revealed no evidence of lymphosarcoma. This case represents a variation of young adult-onset B cell deficiency not previously described in the literature.