Evaluation of serum concentrations of biochemical markers of bone metabolism and insulin-like growth factor I associated with treadmill exercise in young horses

OBJECTIVE: To evaluate changes in serum concentrations of biochemical markers of bone metabolism and insulin-like growth factor I (IGF-I) associated with treadmill exercise in young horses. ANIMALS: 12 two-year-old Thoroughbred mares. PROCEDURE: During a 20-week study period, 6 horses were exercised on a treadmill 3 times a week (exercise group) and 6 horses received walking exercise 6 days a week (controls). Serum concentrations or activity of biochemical markers and IGF-I were assessed biweekly. Bone mineral density and content of the first phalanx were measured by dual-energy X-ray absorbiometry (DEXA) on completion of the study. RESULTS: Compared with values in controls, bone mineral density and content were higher and serum concentrations of osteocalcin (a marker of bone formation) and the carboxy-terminal telopeptide of type I collagen (a marker of bone resorption; ICTP) were lower in exercised horses. Serum concentration and activity of the bone formation markers carboxy-terminal propeptide of type I collagen and bone-specific alkaline phosphatase (BAP) were not different between the 2 groups. Serum IGF-I concentration was lower in the exercise group, compared with control values; there was a significant correlation between change in IGF-I values and changes in osteocalcin, ICTP, and BAP values at the end of the study. CONCLUSIONS AND CLINICAL RELEVANCE: Treadmill exercise over 20 weeks induced adaptive changes in bones of 2-year-old Thoroughbreds; training appears to increase bone mineral density, thereby enhancing mechanical strength of bone, but decreases bone turnover. Results indicated an association between changes in serum IGF-I concentration and bone cell activity in horses.     

Parathyroid hormone (1-34) improves bone mineral density and glucose metabolism in Spontaneously Diabetic Torii-Lepr(fa) rats

The Spontaneously Diabetic Torii-Lepr(fa) (SDT-fa/fa) rat, a model of obese type 2 diabetes, shows obesity, hyperglycaemia and low bone mineral density (BMD). The objective of this study is to evaluate the effects of parathyroid hormone (1-34) [PTH(1-34)] on BMD and glucose metabolism in the SDT-fa/fa rat. SDT-fa/fa rats showed obesity with hyperglycaemia and decreased serum osteocalcin levels and the tibial BMD. A 4-week treatment of PTH(1-34) (20 μg/kg/day) increased the serum osteocalcin levels and the tibial BMD, and decreased the serum glucose levels without changing the serum insulin levels. These findings indicate that PTH(1-34) improved not only BMD but also glucose metabolism in SDT-fa/fa rats. This study suggests that PTH(1-34) is a novel agent for the treatment of diabetic osteoporosis.     

Influence of different calcium concentrations in the diet on bone metabolism in growing dairy goats and sheep

The purpose of this study was to investigate, if different Ca concentrations in diets have an influence on bone mineral metabolism in growing goats and sheep. Twelve growing goats and sheep were divided into two groups. The two control groups received 6.1 g calcium/day (nG) and 6.7 g calcium/day (nS) for goat and sheep respectively. The other two groups were fed 17.7 g calcium/day (hG) and 18.5 g calcium/day (hS). Blood samples were taken 2, 4, 5 and 6 weeks after the start of the experiment. In serum Ca and vitamin D were determined and bone metabolism was measured using crosslinked carboxyterminal telopeptide of type I collagen (ICTP), crosslaps, bone-specific alkaline phosphatase and osteocalcin (OC). Bone mineral density (BMD) was quantified using quantitative computed tomography. Bone resorption marker (ICTP) concentrations were significantly different between both groups control sheep/control goat and hS/hG, but no significant differences were evident in the different feeding groups within one species. OC concentrations showed a similar course to ICTP. The goats had significantly higher concentrations compared with sheep. The 1,25 dihydroxyvitamin D (VITD) concentrations in both hCa groups were significantly lower than in the control groups. BMD increased in the hCa groups compared with the control groups with the time, but significant differences were only evident in sheep in week 2. The hCa diet did not induce differences between the groups within one species for all bone markers. The control Ca diet seems to improve the active Ca absorption via VITD whereas the hCa diet leads to a higher amount of Ca apparently digested. Higher BMD was only observed in group hS compared with nS.     

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.     

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.     

Evaluation of serum osteocalcin and CTX-I in Ardenner horses with special reference to juvenile interphalangeal joint disease

The first aim of this study was to establish a profile of age-related normal serum concentrations of osteocalcin (OC) in Ardenner horses. For this first part, blood samples from 49 healthy Ardenner horses were collected. The second aim was to study two biochemical markers of bone metabolism, OC and a carboxy-terminal cross-linking telopeptide of type I collagen (CTX-I), in 30 young Ardenner horses during 1 year. Amongst them, 17 showed lesions of juvenile degenerative joint disease in the distal forelimbs. A specific radioimmunoassay for equine OC was used to measure the serum concentration of the marker. The serum concentration of CTX-I was measured using a commercially available human assay validated for use in the horse. The effect of age, sex, season and health status (with or without lesions) was assessed. Levels of OC fall between birth and the adult stage: this decrease being most marked between birth and 1 year of age. This age-related decrease of OC was confirmed in the 30 young Ardenner horses, but CTX-I levels remained constant in this group. The Levels of the two markers changed significantly with the season with higher concentrations during the winter. No significant difference was shown either between the two sexes or between the two health statuses.     

Validation and clinical utility of a novel immunoradiometric assay exclusively for biologically active whole parathyroid hormone in the horse

REASONS FOR PERFORMING STUDY: Parathyroid hormone (PTH) plays a critical role in the regulation of mineral metabolism in mammals. Until recently, the standard method for PTH measurement has been the 2nd generation intact-PTH (I-PTH) assay. Current evidence indicates that the I-PTH assay binds to the PTH molecule and to an inactive N-terminally truncated PTH fragment that tends to accumulate in the blood of uraemic patients. Therefore, a new 3rd generation PTH assay that detects only the whole PTH molecule (W-PTH; cyclase-activating PTH [CAP]) has been developed. OBJECTIVES: To validate this more specific W-PTH assay for measurement of equine PTH and evaluate its clinical utility. METHODS: W-PTH and I-PTH were measured in plasma samples from normal horses (adults and foals) and horses with nutritional secondary hyperparathyroidism (N2HPT) and with chronic renal failure (CRF). Replicate measurements and dilutional paralellism were used for assay validation. Changes in blood ionized calcium were induced by EDTA and CaCl2 administration. RESULTS: Performance of the W-PTH assay (accuracy, sensitivity, specificity and ability to detect changes in PTH in response to changes in calcium) was similar to that of the I-PTH assay. Surprisingly, the relative W-PTH concentration in normal horses and foals was higher than the relative I-PTH concentration. W-PTH values remained higher than I-PTH during acute hypo- and hypercalcaemia. An increase in both W-PTH and I-PTH concentrations was found in horses with N2HPT. In horses with CRF, W-PTH and I-PTH values were very low and no increase in I-PTH was observed. CONCLUSIONS: The W-PTH assay can be used for measurement of equine PTH. POTENTIAL RELEVANCE: The use of W-PTH assay is likely to improve the diagnosis of mineral metabolism in horses.     

Radioimmunoassay for parathyroid hormone in equids

Radioimmunoassay for parathyroid hormone (PTH) in equids was performed on blood samples from healthy equids and equids with hypercalcemia and hypophosphatemia. The assay was validated for equine carboxy-terminal PTH. Manipulation of serum ionized Ca in healthy equids by infusing Na2 EDTA and CaCl2 produced an expected increase and decrease, respectively, in measurable immunoreactive PTH. Intra-assay and interassay coefficients of variation were 2.6% and 11.7%, respectively. The range of PTH valves for healthy mature horse mares and geldings maintained on pasture was less than 0.27 ng/ml to 0.92 ng/ml and for horse colts fed grain was 0.61 to 1.25 ng/ml. Serum PTH values were measured on 2 equine patients with hypercalcemia, 1 pony with primary hyperparathyroidism and 1 horse with pseudohyperparathyroidism. Both patients had increased serum PTH values.     

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.     

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.     

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.     

Effects of continuous oral administration of phenylbutazone on biomarkers of cartilage and bone metabolism in horses

OBJECTIVE: To evaluate the effects of continuous oral administration of phenylbutazone on serum and synovial fluid biomarkers of skeletal matrix metabolism in horses. ANIMALS: 11 adult female horses without clinical or radiographic evidence of joint disease. PROCEDURES: Horses were randomly assigned to control or treatment groups. Phenylbutazone was administered orally twice daily at a dose of 4.4 mg/kg for 3 days to the treatment group and subsequently at a dose of 2.2 mg/kg for 7 days. Serum and radiocarpal synovial fluid samples were obtained at baseline and thereafter at regular intervals for 4 weeks. Biomarkers of cartilage aggrecan synthesis (chondroitin sulfate 846) and type II collagen synthesis (procollagen type II C-propeptide) and degradation (collagen type II cleavage) were assayed. Biomarkers of bone synthesis (osteocalcin) and resorption (C-terminal telopeptide of type I collagen) were also measured. RESULTS: No significant differences were found between control and treatment groups or temporally for the biomarkers chondroitin sulfate 846, procollagen type II C-propeptide, collagen type II cleavage, and C-terminal telopeptide of type I collagen in serum or synovial fluid. A significant increase in osteocalcin concentration occurred in synovial fluid during treatment in the treated group. No treatment effect was detected for serum osteocalcin concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that continuous phenylbutazone administration at recommended doses altered some biomarkers in healthy equine joints after short periods of administration. Increased osteocalcin concentration may indicate an undetermined anabolic effect of phenylbutazone administration on periarticular bone or transient induction of osteogenesis in articular chondrocytes or a mesenchymal subpopulation of synoviocytes.     

Urinary markers of type I collagen degradation in the dog

Urinary assays for type I collagen metabolites provide a non invasive index of bone resorption in humans, and are widely used in the management of patients with metabolic bone diseases. The specific aims of this study were to investigate the feasibility of using commercial human assay kits for quantifying the urinary excretion of type I collagen metabolites in dogs of different ages. Urine and serum samples were collected from 35 beagle dogs in five age groups (0 to 1 years; 1 to 2 years; 2 to 3 years; 3 to 7 years; > 8 years old). Urinary concentrations of pyridinoline (Pyd), deoxypyridinoline (Dpd), and the carboxy- and amino-terminal cross-linked telopeptides of type I collagen (CTx and NTx, respectively) were measured with commercial enzyme-linked immunoassay kits. Serum concentrations of another type I collagen metabolite, the carboxy-terminal cross-linked teloptide of type I collagen (ICTP), were measured with a commercial radioimmunoassay. Dilutional studies indicated that the four urinary assays show specific cross-reactivity with canine urine. Age-related differences in urinary marker excretion were identified, with young dogs excreting the highest concentrations of Pyd, Dpd, NTx and CTx. The correlation between the individual urinary markers was excellent (r = 0.87 to 0.98), while the correlation between serum ICTP and individual urinary markers was weaker (r = 0.52 to 0.64). These results validate the usefulness of the commercial assay kits in monitoring type I collagen metabolism in dogs. Histomorphometric studies have confirmed the relationship between collagen degradation and bone resorption in humans, and similar studies are now needed in dogs.     

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.     

Effects of storage on serum ionized calcium and pH from horses with normal and abnormal ionized calcium concentrations

It has been previously shown that Ca(I) concentration is stable in serum collected from healthy horses for 10 days if stored at 40 degrees C. This may not be true for horses with abnormal Ca(I) concentrations. Thus the stability of ionized calcium (Ca(I)) concentration and pH measurement in serum from horses with both normal and abnormal Ca(I) concentrations stored for various times at 40 degrees C and -10 degrees C was evaluated. Our results indicated that serum Ca(I) concentration was stable throughout 7 days of cold or frozen storage, after being received by the Clinical Chemistry Laboratory. Serum Ca(I) concentration showed a significant decrease by 14 days of frozen storage (-10 degrees C). Serum pH showed a statistically significant increase by 7 days of cold storage, and within 3 days of frozen storage. If equine serum is collected, handled and stored anaerobically, and kept cold or frozen, Ca(I) concentration can be accurately measured for approximately 7 days after collection, regardless of the health status of the animal. An accurate measurement of pH may be made within 3 days of cold or 1 day of frozen storage.     

Influence of a vegetarian diet versus a diet with fishmeal on bone in growing pigs

This study was conducted to examine if substantial bone loss occurs in growing pigs fed a vegetarian diet in comparison with a diet containing fishmeal. Twelve 6-week-old weaned pigs were assigned to two groups: group V [vegetarian diet; 0.61% phosphorus (P) in dry matter until 25 kg and 0.46% P until the end of the experiment] and group F (fishmeal diet; 0.61% P in dry matter until 25 kg and 0.46% P until the end of the experiment). Phytase was added to both diets. These two diets were fed to the two groups for a period of 6 weeks. Blood samples were collected weekly, faeces were collected three times a week. Concentrations of osteocalcin (OC) and carboxyterminal telopeptide of type I collagen (ICTP) were measured in serum, using a radioimmunoassay, and bone-specific alkaline phosphatase (bAP) was measured using an enzyme immunoassay. Bone mineral density (BMD) and content (BMC) were determined by peripheral quantitative computer tomography (pQCT) in the tibia and phalanx. In addition, 1,25-dihydroxyvitamin D (VitD) and parathyroid hormone (PTH) were measured in serum. The digestibility of P was significantly decreased in group V. Significant changes in bAP activities and OC concentrations occurred with time during the 6 weeks. ICTP concentrations were significantly higher in group V. Total BMC and BMD in the tibia and BMD in the phalanx significantly decreased in group V. The results show that a vegetarian diet induces a significant loss of bone and a higher bone formation in group V compared with group F, although phytase was added to both diets. The dietary requirements for P in pigs, especially in the context of feeding vegetarian diets and adding an appropriate amount of phytase, should be investigated further.     

The effect of equine recombinant growth hormone on second intention wound healing in horses

OBJECTIVE: To evaluate the effect of intramuscular administration of recombinant equine growth hormone on healing of full thickness skin wounds on equine limbs. STUDY DESIGN: Experimental. ANIMALS: Nine Standardbred horses. METHODS: In study 1, standardized full thickness skin wounds (2.5 x 2.5 cm) were made over the dorsomedial aspect of the mid-cannon bone of 1 forelimb and 1 hindlimb in 9 horses. Wounds were bandaged without treatment (control subjects) and videorecorded twice weekly until healed. Then, in study 2, similar wounds were created on the opposite limbs; 6 horses were administered intramuscular recombinant equine growth hormone (10 microg/kg daily for 7 days, then 20 microg/kg daily for 49 days), and 3 horses (control subjects) were administered equivalent volumes of sterile water. Wounds were videorecorded twice weekly until healed. Wound healing variables were measured from the videorecordings using a computer software package and analyzed as a randomized complete block design factorial analysis of variance; significance was set at P <.05. RESULTS: No differences in the measured variables were detected between wounds in study 1 and the control wounds in study 2. In recombinant equine growth hormone-treated horses, wounds retracted more during treatment and contracted faster after treatment stopped when compared with wounds from untreated horses. No other treatment effects were detected. CONCLUSIONS: Recombinant equine growth hormone seemingly increases wound retraction. After treatment ceases, wound contraction increases. CLINICAL RELEVANCE: Intramuscular administration of recombinant equine growth hormone (10 microg/kg daily for 7 days, then 20 microg/kg daily for 49 days) does not appear to have any beneficial clinical effect on healing of equine limb wounds.     

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.     

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.     

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.