Response of serum biochemical markers of bone metabolism totraining in the juvenile racehorse

Sixteen Quarter Horse-type geldings were used to examine the response of biochemical markers of bone metabolism to forced exercise prior to and during race training. The study began when the average age of the horses was 15 months. Horses were exercised on a high-speed treadmill for 14 weeks, and were subsequently placed into race training. Serum was collected and assayed for concentrations of osteocalcin (BGP), the carboxyterminal telopeptide of type I collagen (ICTP) and the carboxyterminal propeptide of type I procollagen (PICP). When data were normalized from the onset of race training, ICTP and PICP concentrations were higher in the pre-exercised horses (P<.05 and P<.1, respectively) indicating higher rates of bone turnover. Overall, bone turnover appeared to be decreased during race training, as concentrations of PICP and ICTP were lower when compared to values seen during the pre-training Phase.     

Serum osteocalcin and CTX-MMP concentration in young exercising thoroughbred racehorses

Bone responds to exercise with changes in bone (re-)modelling, which might be monitored non-invasively with biochemical bone markers. The aim of this study was to evaluate the influence of exercise on serum osteocalcin and serum carboxy-terminal cross-linked telopeptide of type I collagen generated by matrix metalloproteinases (CTX-MMP) concentration in young racehorses. Seventy-one 2 to 4-year-old Thoroughbreds were included in this prospective infield study. Blood sampling was performed six times (i.e. six sampling cycles) during a 9-month period. Serum samples were analysed with commercial osteocalcin and CTX-MMP radioimmunoassays. Two-year-old racehorses had higher serum osteocalcin and CTX-MMP values than 3-year-old horses. Gender and training amplitude did not significantly influence serum osteocalcin and CTX-MMP values. Two-year-old horses showed an increase in osteocalcin values between cycles 2 and 3 and an increase in serum CTX-MMP values between cycles 1 and 2. Serum osteocalcin and CTX-MMP concentrations decreased between cycles 4 and 5, and 5 and 6. Three-year-old horses showed an increase in serum osteocalcin levels between cycles 3 and 4 and an increase in serum CTX-MMP concentrations between cycles 1 and 2, and 3 and 4. Serum osteocalcin levels decreased between cycles 5 and 6, whereas serum CTX-MMP levels decreased between cycles 4 and 5, and 5 and 6. Two- and three-year-old horses showed a decreased osteocalcin/CTX-MMP ratio between cycles 1 and 2. Moreover, 2-year-old horses showed an increase in the osteocalcin/CTX-MMP ratio between cycles 2 and 3. Sore shin formation did not significantly influence serum osteocalcin and CTX-MMP values. Serum osteocalcin and CTX-MMP are promising bone markers for monitoring exercise induced changes in equine bone metabolism.     

Biochemical markers of bone activity in young standardbred horses during different types of exercise and training

Seven untrained Standardbred horses were used in a training programme of 6 weeks to evaluate the effects of exercise and training on bone metabolism. The horses were exercised on a treadmill according to a standardized exercise test (SET 1: six incremental steps, 5 min duration each; start 5 m/s, increase 1 m/s). SET 1 was followed by a training programme of 6 weeks. In alternating order: high-speed exercise (HSE): 15 min duration, start at VLa4, continuous increase in speed every 60 s by 0.3 m/s (14 incremental steps); low-speed exercise (LSE): constant velocity at VLa2.5, duration: approximately 60-90 min (total training programme: eight HSE and eight LSE sessions). SET 2 finished the training programme and a deconditioning period of 12 weeks followed. Blood samples for lactate, total plasma protein (TPP), osteocalcin, and ICTP (cross-linked C-telopeptide of type I collagen) were collected. ICTP increased during SET 1 and SET 2, whereas osteocalcin decreased to below resting concentration 24 h after SET 1. A rise in ICTP was observed during LSE 1 and LSE 8, which was followed by a drop 24 h after exercise. No changes in osteocalcin were noted during LSE 1, but 24 h after LSE 1 osteocalcin dropped to below pre-exercise levels. LSE 8 resulted in an increase in osteocalcin, followed by a drop 24 h after LSE 8. Osteocalcin and ICTP were not affected by HSE. Baseline osteocalcin levels dropped during the course of training. The acute response of biochemical bone markers indicates a direct influence of a single bout of exercise on bone metabolism.     

Biochemical markers of bone metabolism in growing thoroughbreds: a longitudinal study

This study describes longitudinal changes in serum levels of biochemical markers of bone cell activity in a group of 24 thoroughbred foals from birth to 18 months of age. The markers of bone formation included the type I collagen carboxy-terminal propeptide (PICP), the bone-specific isoenzyme of alkaline phosphatase (BAP), and osteocalcin (OC). Levels of the cross-linked telopeptide of type I collagen (ICTP), a marker of bone resorption, and the N-terminal propeptide of type III collagen (PNIIIP), a marker of soft tissue turnover, were also measured. Levels of all markers fell significantly between birth and 18 months of age (70-80 per cent); this decrease being most marked between 0 and 6 months. However, a transient increase in levels of the markers then occurred between 6 and 14 months of age. The timing of this increase was specific for each parameter. ICTP and OC concentrations increased between October and December. PICP concentrations increased between December and April whereas the increase in PIIINP was coincident with the peak in weight gain between April and June. Changes in BAP concentration were less distinct at this time. Season was shown to have significant effects on the biochemical markers independent from the effect of age. Concentrations of all markers decreased with increasing body weight and at any given age heavier horses had lower marker levels. These results show that biochemical markers of bone cell activity and soft tissue turnover follow characteristic patterns of change in growing thoroughbreds influenced by age, season and bodyweight. The demonstration that the reference ranges for the biochemical markers change from month to month means that single samples from individuals are of little value for monitoring bone cell activity in growing thoroughbreds.     

Relationship between stages of the estrous cycle and bone cell activity in Thoroughbreds

OBJECTIVE: To investigate the relationship between stage of estrous cycle and bone cell activity in Thoroughbreds. SAMPLE POPULATION: Blood samples collected from forty-seven 2-year-old Thoroughbred mares in training for racing. PROCEDURES: Blood samples were collected monthly (in April through September) from the mares. Stage of estrus was determined by assessing serum progesterone concentration. Bone cell activity was determined by measuring concentrations of 2 markers of bone formation (osteocalcin and the carboxy-terminal propeptide of type I collagen [PICP]) and a marker of bone resorption (the cross-linked carboxy-terminal telopeptide of type I collagen [ICTP]) in sera. RESULTS: When the relationship between stage of the estrous cycle and markers of bone cell activity was examined, serum concentrations of both osteocalcin and ICTP were significantly higher in mares that were in the luteal phase, compared with mares that were at other stages of the estrous cycle. Stage of estrus did not affect serum PICP concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that bone cell activity in Thoroughbred mares fluctuates during the estrous cycle; serum concentrations of markers of bone formation and bone resorption are increased during the luteal phase. Further studies are required to determine whether these changes are of clinical importance and increase the risk of injury for mares in training during the breeding season. As in humans, stage of estrus must be considered as a source of uncontrollable variability in serum bone marker concentrations in horses.     

Effect of Treadmill Exercise and Hydrogen-rich Water Intake on Serum Oxidative and Anti-oxidative Metabolites in Serum of Thoroughbred Horses

The present study aimed to clarify changes of oxidative stress and antioxidative functions in treadmill-exercised Thoroughbred horses (n=5, 3 to 7 years old), using recently developed techniques for measurement of serum d-ROMs for oxidative stress, and BAP for antioxidative markers. Also, the effect of nasogastric administration of hydrogen-rich water (HW) or placebo water preceding the treadmill exercise on these parameters was examined. Each horse was subjected to a maximum level of treadmill exercise in which the horses were exhausted at an average speed of 13.2 ± 0.84 m/sec. Blood samples were taken 4 times, immediately before the intake of HW or placebo water at 30 min preceding the treadmill exercise, immediately before the exercise (pre-exercise), immediately after the exercise (post-exercise) and at 30 min following the exercise. In all horses, both d-ROMs and BAP values significantly increased at post-exercise. The increase in d-ROMs tended to be lower in the HW trial, as compared to the placebo trial at pre-exercise. The increase in BAP was considerable at approximately 150% of the pre-exercise values in both the HW and placebo treatment trials. The BAP/d-ROMs ratio was significantly elevated at post-exercise in both treatment trials, while a significant elevation was also observed at pre-exercise in the HW trial. BAP, d-ROM, and the BAP/d-ROM ratio tended to decline at 30 min after the exercise, except BAP and BAP/d-ROMs in the placebo trial. These results demonstrate that the marked elevation of oxidative stress and anitioxidative functions occurred simultaneously in the intensively exercised horses, and suggest a possibility that HW has some antioxidative efficacy.     

Bone Metabolism Markers in Arabian Horses During the First Two Years of Life

Serum markers of bone metabolism were analyzed in Arabian horses from birth through 2 yr. The marker of bone formation utilized was osteocalcin (OC), and the marker for degradation was carboxy-terminal pyridinoline cross-linked telopeptide region of type I collagen (ICTP). Blood samples were taken via jugular venipuncture the day of birth, d 15, d 30, d 45, d 60, and every 30 d thereafter through d 720. Serum was obtained and analyzed for OC and ICTP. Osteocalcin concentrations increased immediately after birth, were variable, and returned to baseline by d 300. By d 330, concentrations of OC began to decrease from d 0 and stayed at this lower concentration through d 510. From d 540 through 720, OC concentrations were similar to baseline. A decrease from baseline (d 0) in ICTP concentrations was seen on d 60, and ICTP continued to decline in concentration through d 720. Therefore, concentrations of OC and ICTP decreased over time as previously reported, and this study characterizes those changes on a monthly basis. Variability and general concentrations for OC and ICTP obtained in this study will provide valuable information for future experimental design and use of these markers in young horses and will aid researchers in determining treatment effects without being confounded by changes in concentrations caused by growth.     

Biochemical Markers of Bone Modeling and Remodeling in Juvenile Racehorses at Varying Mineral Intakes

In this study, blood-borne biochemical markers were used to track comparative rates of bone turnover in horses fed differing amounts of Ca, P and Mg. Bone turnover was tracked by serum osteocalcin; bone resorption by the carboxyterminal telopeptide of type I collagen (ICTP); and bone formation by the carboxyterminal propeptide of type I procollagen (PICP). Twenty-one longyearling Quarter Horses were blocked by gender and age, randomly assigned to one of four diets and subjected to 128 d of race training. The study was conducted in 32-d periods, each consisting of 28 d of race training followed by a 4-d fecal and urine collection, or a 4-d rest period. Blood samples were taken weekly during the training period. Serum and plasma samples were analyzed for biochemical markers of bone activity using RIA procedures. Onset of training resulted in elevated blood concentrations of ICTP, PICP and osteocalcin. Concentrations of ICTP and PICP were greater during the first 64 d of training, indicating increased bone activity during the first half of the training period. Horses with the greatest intake of minerals exhibited greater concentrations of PICP (bone formation marker) and lesser concentrations of ICTP (bone breakdown marker). Further, ICTP, PICP and osteocalcin concentrations decreased dramatically following 4-d of confinement and relative inactivity. Therefore it appears that feeding minerals at levels greater than current NRC recommendations provided a protective effect on the developing skeleton of the young racehorse. Additionally, the biochemical markers used in this study were sensitive enough to track daily changes in bone activity resulting from daily changes in stress to the skeleton.     

Long-term effects of intermittent equine parathyroid hormone fragment (ePTH-1-37) administration on bone metabolism in healthy horses

Intermittent administration of parathyroid hormone (PTH) is an anabolic therapy for osteoporotic conditions in humans. This study evaluated the effects of equine PTH fragment (ePTH-1-37) administration on bone metabolism in 12 healthy horses. Six horses each were treated once daily for 120days with subcutaneous injections of 0.5μg/kg ePTH-1-37 or placebo. Blood was collected to determine ionized calcium (Ca(++)), total Ca (Ca(T)), inorganic phosphorus, serum equine osteocalcin (eOC), carboxy-terminal telopeptide of type I collagen (ICTP), bone-specific alkaline phosphatase, and carboxy-terminal cross-linked telopeptide of type I collagen. Bone mineral density (BMD) was determined with dual X-ray absorptiometry of the metacarpus and calcaneus. Significantly higher blood Ca(++) and plasma Ca(T) concentrations were measured 5h after ePTH-1-37 administration compared to placebo. Higher serum eOC concentrations were found for ePTH-1-37 treatment at days 90 (P<0.05) and 120 (P=0.05). Significantly higher serum ICTP levels were observed with ePTH-1-37 treatment at days 60 and 90. For both study groups, BMD increased significantly in the calcaneus. Long-term use of ePTH-1-37 seemed to have no negative effects on bone metabolism in healthy horses. The absence of undesirable side effects is the premise to ensure safety for further clinical investigations in horses with increased bone resorption processes.     

The effect of initiation of exercise training in young horses on vitamin K status

Twelve Quarter Horses, 18 to 24 mo of age and having an average body weight of 460+/-12 kg, were randomly assigned to one of two exercise treatments (control and exercise) to study the effect of initiation of exercise training in young horses on vitamin K status. The control treatment consisted of hand-walking the horses 7 min/d and 5 d/wk for 180 d. Exercise consisted of the following treadmill exercise regimen: 2 min at 4.5 to 5.4 m/s, 3 min at 13.4 to 14.3 m/s, and 2 min at 4.5 to 5.4 m/s on 5 d/wk for 180 d. Both groups were allowed free access to brome grass hay (273 mg of phylloquinone/100 g) individually. Additionally, the exercise group was fed .45 kg of a grain-mix concentrate (40 mg of phylloquinone/100 g) on the days they were exercised. Jugular venous blood samples were collected at d 0, 30, 60, 90, 120, 150, and 180. Blood samples were analyzed for total serum osteocalcin and vitamin K status via the hydroxyapatite binding capacity of serum osteocalcin calculated as follows: Hydroxyapatite binding capacity of serum osteocalcin = [(total serum osteocalcin - serum osteocalcin following extraction of serum with hydroxyapatite)/total serum osteocalcin] x 100. All horses were killed with an overdose of sodium pentobarbital on d 180. Computed tomographic osteoabsorptiometry was used to measure relative bone density distribution on the surface of the distal radial carpal bone, proximal third carpal bone, and the distal third metacarpal condyle. Relative bone density distribution was measured in Hounsfield units and categorized as low-, medium-, and high-density bone corresponding to 800 to 1,199, 1,200 to 1,299, and 1,300 to 3,000 Hounsfield units, respectively. Carpal and metacarpophalangeal joints were assigned a score of 0 (normal) to 3 (severe) that reflected the presence and severity of joint lesions. Hydroxyapatite binding capacity of serum osteocalcin and serum osteocalcin were unchanged over the 180-d period in both groups. Exercised horses had a higher percentage of high density bone (P < .01) and a lower percentage of low density bone (P < .01) on the surface of the third metacarpal condyle and a higher cumulative gross lesion score (P < .01) in the metacarpophalangeal joint than controls. There were no differences between treatments in relative bone density distribution in the carpal bones, or cumulative gross lesion score of the carpal joints. No significant correlation was present between the serum measures, osteocalcin and hydroxyapatite binding capacity of serum osteocalcin, and the bone measures, relative bone density distribution and joint gross lesion score.     

Clinical effects of exercise on subchondral bone of carpal and metacarpophalangeal joints in horses

OBJECTIVE: To determine effects of treadmill exercise on subchondral bone of carpal and metacarpophalangeal joints of 2-year-old horses. ANIMALS: 12 healthy 2-year-old horses. PROCEDURE: Horses were randomly assigned to the control (n = 6) or exercised (6) groups. Horses in the exercised group ran on a high-speed treadmill 5 d/wk for 6 months. Horses in the control group were hand walked for the same amount of time. Results of clinical, radiographic, nuclear scintigraphic, and computed tomographic examinations, and serum and synovial concentrations of biochemical markers of bone metabolism were compared between groups. RESULTS: Exercised horses were significantly lamer at the end of the study than control horses. Radionuclide uptake in the metacarpal condyles, but not in the carpal joints, was greater in exercised horses, compared with control horses. Exercised horses also had a higher subchondral bone density in the metacarpal condyles than control horses, but such differences were not detected in the carpal bones. CONCLUSIONS AND CLINICAL RELEVANCE: None of the diagnostic techniques evaluated was sufficiently sensitive to detect all osteochondral damage. Computed tomography and computed tomographic osteoabsorptiometry were superior to conventional radiography for detecting small osteochondral fragments. Nuclear scintigraphy was a sensitive indicator of subchondral bone change but lacked specificity for describing lesions and discerning normal bone remodeling from damage. Newer techniques such as computed tomography may help clinicians better diagnose early and subtle joint lesions in horses prior to development of gross joint damage.     

Response of Biochemical Markers of Bone Metabolism to Exercise Intensity in Thoroughbred Horses

We studied the response of biochemical markers of bone metabolism to exercise intensity in horses. Four horses were walked on a mechanical walker for one week (pre-exercise). Then they performed low-speed exercise on a high-speed treadmill in the first week and medium-speed exercise in the second week and high-speed exercise in the third week of training. We measured two indices of bone resorption, serum hydroxyproline concentration and the urinary deoxypyridinoline/creatinine ratio, and serum osteocalcin (OC) concentration as an index of bone formation. Both indices of bone resorption gradually decreased during the experiment. Serum OC concentration did not change in the first week but was significantly lower in the second and the third weeks compared to in the pre-exercise period and in the first week. These results suggest that the low-speed exercise decreased bone resorption but did not affect bone formation, which possibly results in increasing bone mineral content and strengthening of bones. The high-speed exercise decreased bone formation and bone resorption, i.e., bone turnover was suppressed. The low-speed exercise may be preferable for increasing bone mineral content.     

Circadian variation in biochemical markers of bone cell activity and insulin-like growth factor-I in two-year-old horses

Studies in humans have found circadian changes to be one of the most important sources of controllable preanalytical variability when evaluating bone cell activity using biochemical markers. It remains unclear whether similar circadian changes influence bone marker concentrations in the horse. The aim of this study was to characterize changes in serum concentrations of three biochemical markers of bone cell activity over a 24-h period in six 2-yr-old Thoroughbred mares, and to determine circadian variability in IGF-I, which regulates bone turnover. Three bone markers were measured in serum: osteocalcin, a marker of bone formation, the carboxy-terminal propeptide of type-I collagen (a marker of bone formation), and the carboxy-terminal telopeptide of type-I collagen (a marker of bone resorption). Data were analyzed using the cosinor technique, which fits a 24-h cycle to each dataset. A significant circadian rhythm was observed for osteocalcin (P = 0.028), with an estimated amplitude of 7.6% of the mean (95% confidence interval 1.3% to 16.3%), and an estimated peak time of 0900. However, the observed rhythm for the carboxy-terminal telopeptide of type-I collagen (amplitude = 7.4%) was not significant (P = 0.067), and there were no significant changes in concentrations of the carboxy-terminal propeptide of type-I collagen over the 24-h study period (P = 0.44). There was a small but significant circadian rhythm for IGF-I (P = 0.04), with an estimated amplitude of 3.4% (95% confidence interval 0.2 to 7.1%) and peak at 1730. Further studies are now required to determine the potential association between circadian changes in IGF-I and osteocalcin in the horse. Although no significant circadian variation was found in concentrations of the car-boxy-terminal propeptide of type-I collagen and the carboxy-terminal telopeptide of type-I collagen, this may in part be a result of the age of the animals that were still skeletally immature. Future studies should aim to determine whether these markers develop a circadian rhythm at a later age when growth is complete. In the meantime, consistency in time of sampling should continue to be considered best practice when measuring biochemical markers of bone turnover in the horse.     

Markers of bone metabolism in dog breeds of different size

Serum and urinary markers of bone turnover may be of value in animals as noninvasive tools for determining the response of the skeleton to disease and injury. Although normal values for bone markers have been reported for the Beagle, concerns remain that breed to breed differences will complicate the interpretation of bone marker data in dogs. To explore this, we examined serum bone markers in two breeds of vastly different size, Pomeranians and Irish Wolfhounds. Our hypothesis was that serum concentrations of bone markers are similar in toy and giant dog breeds and fall within the same range as those reported for Beagles. Bone alkaline phosphatase (BALP) and carboxy-terminal telopeptide of type I collagen (ICTP), respectively markers of bone formation and bone resorption, were measured in age matched Pomeranians (n=14) and Irish Wolfhounds (n=14). No statistically significant differences between the mean BALP and mean ICTP serum concentrations from Pomeranians and Irish Wolfhounds were found. All BALP and ICTP concentrations were within the reference range reported for Beagles. The results of this study suggest that serum BALP and ICTP concentrations in giant and toy breeds are the same as in Beagles and that these assays may be used for dogs of all sizes.     

Evaluation of plasma carboxy-terminal cross-linking telopeptide of type I collagen concentration in horses

OBJECTIVE: To evaluate a human assay for quantification of carboxy-terminal cross-linking telopeptide of type I collagen (CTX-I), assess the influence of age on plasma CTX-I concentration, investigate the relationship between plasma CTX-I and serum osteocalcin concentrations, and determine whether concentrations of plasma CTX-I or serum osteocalcin fluctuate in circadian manner in horses. HORSES: 75 clinically normal horses. PROCEDURE: Cross-reactivity between equine serum CTX-I and CTX-I antibodies in an automated electrochemiluminescent sandwich antibody assay (ECLIA) was evaluated via a specificity test (ie, dilution test) and recovery calculation. Serum osteocalcin concentration was measured with an equine-specific osteocalcin radioimmunoassay. To analyze diurnal variations in plasma CTX-I and serum osteocalcin concentrations, blood samples were obtained hourly during a 24-hour period. RESULTS: Results of the dilution test indicated good correlation (r > 0.99) between expected serum CTX-I concentrations and measured serum CTX-I concentrations. The calculated CTX-I recovery was 97.6% to 109.9%. Plasma CTX-I and serum osteocalcin concentrations were correlated. Plasma CTX-I concentration was inversely correlated with age of the horse. No significant circadian variations in plasma CTX-I and serum osteocalcin concentrations were detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the fully automated CTX-I ECLIA can be used for evaluation of plasma and serum samples from horses and may be a useful tool to monitor bone metabolism changes. Horses in this study did not have notable diurnal fluctuations in serum osteocalcin and plasma CTX-I concentrations.     

Biochemical markers of bone metabolism in draught andwarmblood horses

Concentrations of the cross-linked carboxyterminal telopeptide of type I collagen (ICTP) and osteocalcin (OC) have been determined in the serum of one hundred clinically healthy adult Draught or Warmblood horses. The correlation between these two markers has been evaluated and the influence of gender, age and type of horse described. No significant variations were observed between animals of different sex, but a significant inverse correlation (P<0.0001) with age was observed for both measured parameters. After correction for age, serum levels of OC were found to be lower in Draught [adjusted least square mean (LSM)=6.612μg. L⁻¹] than in Warmblood horses (adjusted LSM=8.596μg.L⁻¹), whereas levels of ICTP were higher in Draughts (adjusted LSM=8.035pg.L⁻¹) than in Warmbloods (adjusted LSM=6.643μg.L⁻¹). A significant correlation (P<0.0001) was observed between OC and ICTP. This correlation was stronger if the type of horse was taken into account in the statistical model. The ratio OC:ICTP was independent of gender and age. A higher OC:ICTP ratio in Warmbloods compared to the Draught horses might reflect a higher bone remodelling level of horses submitted to regular daily work. It was concluded that ICTP and OC are influenced by the type of horse, and probably reflect a physiological difference in bone remodelling between these animals.     

Biochemical markers of bone metabolism and risk of dorsal metacarpal disease in 2-year-old Thoroughbreds

REASONS FOR PERFORMING STUDY: Dorsal metacarpal disease (DMD) is a common problem in 2-year-old racehorses and results in loss of a significant number of days from training. Biochemical markers of bone cell activity measured early in the training season could have value for identifying 2-year-old Thoroughbred racehorses that develop DMD. OBJECTIVES: To determine the association between serum concentrations of osteocalcin, the carboxyterminal propeptide of type I collagen (PICP) and the carboxyterminal cross-linked telopeptide of type I collagen (ICTP) measured early in the training season and the risk of DMD. METHODS: Blood samples were collected from 165 two-year-old Thoroughbreds during late November/early December. Osteocalcin and PICP were measured as markers of bone formation, and ICTP as a marker of bone resorption. Training and veterinary records for each horse were monitored over the following training/racing season (10 months). Cases were defined as an episode where signs of DMD were sufficiently severe for a horse to miss at least 5 consecutive days of training. Classification tree and logistic regression analysis were used to identify the most important factors suitable for prediction of DMD risk. RESULTS: There were 24 cases of DMD during the season (14.6% cumulative incidence), with an average time to recognition of approximately 6 months (May). The earliest recognised case was in February and the latest in September. Osteocalcin and ICTP concentrations in the early stages of the training season were significantly higher in horses that subsequently developed DMD (P = 0.017 and 0.019, respectively). DMD cases were also significantly older compared to noncases (21.04 vs. 20.44 months, P = 0.023). Using a multivariable logistic regression model, it was possible to postulate a set of diagnostic rules to predict the likelihood of DMD injury during the season. This suggested that horses with ICTP concentrations above 12365 ug/l and older than 20.5 months are 2.6 times more likely to develop DMD. CONCLUSIONS: The measurement of the bone resorption marker ICTP could be useful for identification of 2-year-olds at increased risk of developing DMD. POTENTIAL RELEVANCE: These findings, together with other strategies such as modification of training regimens, e.g. early introduction of short distances of high-speed exercise into the training programme, could help reduce the days lost to training as a result of DMD.     

Influence of age and sex on serum osteocalcin concentrations in horses at weaning and during physical conditioning

Three experiments were conducted to evaluate serum osteocalcin concentrations in normal weanling and yearling Quarter Horses. In Experiment 1, jugular blood samples were taken at 3 hr intervals for 24 hr to evaluate diurnal changes in serum osteocalcin concentration of foals (n=3) and yearlings (n=5). In Experiment 2, twelve Quarter Horse foals were weaned at 4 months of age to determine the influence of sex, weaning and method of weaning of serum osteocalcin concentration. The third experiment utilized fifteen yearling Quarter Horses (7 geldings, 8 fillies) in a two-phase trial to evaluate normal peripheral osteocalcin concentration in sedentary and exercising horses. In Experiment 1, there was no detectable variation (P>.05) in serum osteocalcin concentration during the 24 hr sample period in either age group. In Experiment 2, colts had greater (P<.05) osteocalcin concentrations than fillies at weaning. Method of weaning did not alter mean serum osteocalcin concentration (P>.1). Serum osteocalcin concentrations declined (P<.05) in all horses following weaning but returned to preweaning levels within one week. In Experiment 3, sedentary horses had similar (P>.1) osteocalcin concentrations on d 0, 45 and 90. Serum osteocalcin concentrations of sedentary horses were not affected by sex (P>.1). During exercise, fillies had greater (P<.05) osteocalcin concentrations than geldings. Serum osteocalcin concentrations of all horses declined linearly (P<.0001) during the 90 d exercise period. Osteocalcin may be useful as a tool to assess bone metabolism during growth and physical conditioning in horses. However, variability in serum osteocalcin concentrations due to age, sex and level of activity suggest that these factors must also be considered.     

Increased plasma silicon concentrations and altered bone resorption in response to sodium zeolite a supplementation in yearling horses

This study examined the effect of supplementation of a bioavailable source of silicon (sodium zeolite A) on altering systemic markers of bone metabolism in horses. Twenty yearlings (ten Quarter Horses and ten Arabians) were randomly grouped as silicon (Si) supplemented (S; n=10), in which yearlings consumed 2% of the total diet as a Si-containing supplement, and a second non-supplemented control group (C; n=10). Blood samples were taken on days 0, 15, 30 and 45. Both plasma and serum were collected; the plasma was analyzed for Si concentrations and serum was analyzed for osteocalcin (OC), carboxy-terminal pyridinoline cross-linked telopeptide region of type I collagen (ICTP), and pyridinoline and deoxypyridinoline crosslinks (PYD). Supplemented yearlings had higher plasma Si concentrations than C yearlings by day 15, and remained higher than C yearlings on days 30 and 45 (P < 0.0001 for all days). There were no differences between treatment groups for OC or PYD concentrations (P > .05); however, ICTP concentrations were lower in S yearlings on day 45 when compared to C yearlings (P = .04). Results indicate that sodium zeolite A supplementation (consumed at 2% of the total diet) increases plasma Si concentrations. Furthermore, results indicate that Si-supplemented yearlings may have decreased bone resorption, which may provide for greater net bone formations, as OC concentrations were not different between groups. Unfortunately, systemic markers give no indication as to the quality of the bone that may be formed, and further research in the area of Si supplementation, bone metabolism and bone strength is required to establish conclusive evidence as to the benefits of supplemental Si to the skeletal system.