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.     

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.     

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.     

Diurnal variation and age differences in the biochemical markers of bone turnover in horses

Biochemical markers of bone turnover provide sensitive, rapid, and noninvasive monitoring of bone resorption and formation. Serum concentrations of osteocalcin (OC) reflect rates of bone formation, and urinary concentrations of the pyridinium crosslinks pyridinoline (Pyd) and deoxypyridinoline (Dpd) are specific and sensitive markers of bone resorption. These markers are age-dependent and are used to detect and monitor changes in the rates of bone turnover in a variety of orthopedic diseases in humans and may prove to have similar application in horses. This study examined age differences and diurnal variation in OC, Pyd, and Dpd in eight adult geldings and seven weanling colts. Blood and urine were collected at regular intervals over 24 h. Serum OC and cortisol, and urinary Pyd and Dpd were analyzed. Mean 24-h concentrations of cortisol and all three markers were higher (P<.003) in weanlings than adults. Significant 24-h variation was observed in adult gelding OC, Pyd, and Dpd concentrations (P< .02). Adult OC concentrations were highest between 2400 and 0900; Pyd and Dpd peaked between 0200 and 0800. Similar patterns of bone turnover were observed in weanling values, but they were not significant (P>.17) owing to greater variability between individuals. Cortisol secretion varied (P<.001) over 24 h in both adults and weanlings and, thus, did not seem to be responsible for greater variability in markers of bone turnover between weanlings. These data demonstrate that diurnal rhythms exist for serum OC and urinary Pyd and Dpd in adult horses, as reported in humans, and that sample timing is an important consideration in future equine studies using these markers.     

Serum levels of the immunoglobulins IgG and IgG(T) in horses

Levels of the immunoglobulins IgG and IgG(T) in serum in Norwegian horses of the breeds “Døle” and “Fjord” were determined by the quantitative radial immunodiffusion test.No significant differences were apparent between the 2 Norwegian breeds. The immunoglobulin levels were approximately in the same range as previously reported for Shetland ponies.Immunoglobulins could not be detected in the newborn foal. As early as 24 hrs. after birth the mean immunoglobulin level was within the adult range. After a drop during the first month of life, the immunoglobulins increased. IgG(T) rose more rapidly and to a higher level than IgG.In 2 year old horses, IgG(T) was significantly higher than in adults, while IgG was significantly lower. IgG(T) seems to be a very important immunoglobulin in foals and young 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.     

Neutrophil functions and serum IgG in growing foals.

The aim of this study was to investigate the phagocytic and killing capacities as well as expression of CD18 of neutrophils obtained from healthy foals from birth to age 8 months. Blood was taken from 6 Standardbred foals at 7 time-points between ages 2-56 days and thereafter once a month. For comparison, cells from 16 mature horses were evaluated. Neutrophil phagocytosis of yeast cells was assessed by flow cytometry after opsonisation with mature pooled serum, autologous serum or anti-yeast IgG. The killing capacity of the neutrophils, as indicated by the oxidative burst, was monitored by chemiluminescence. Serum IgG concentration was measured by radial immunodiffusion. In addition to clinical examination, the amount of serum amyloid A and the total leucocyte count were used as markers for infection. The phagocytic ability was impaired until age 3 weeks, when autologous serum was used as opsonin. Killing capacity was also low initially but, from 3 months onwards, chemiluminescence values were equal to or higher than in mature horses. Serum IgG decreased from 10 g/l at 2 days to 5 g/l at 2 months and then increased gradually to 10 g/l at the end of the study. These findings may in part explain the increased susceptibility to bacterial infections in young horses.     

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.     

Articular / epiphyseal osteochondrosis in Thoroughbred foals at 5 months of age: influences of growth of the foal and prenatal copper supplementation of the dam

AIMS: To determine the influence of copper (Cu) supplementation by injection of mares in late gestation on the frequency and severity of osteochondrosis (OC) lesions in their foals at around 160 days of age. To determine if there was any influence of the concentration of Cu in the liver, growth rate, birthweight, weight at 160 days of age, fatness, sex, or year of birth of the foal on the frequency and severity of OC lesions. To determine the influence of dam's age, and sex and birthweight of the foal on the growth rate from birth to 160 days of age, and weight at 160 days of age. METHODS: Thirty-three Thoroughbred foals, born in two consecutive years, were weighed every 2 weeks from birth. The dams had been supplemented with parenteral Cu or saline during late gestation, and the supplementation regimens were different in each year. Foals had liver biopsies harvested at birth for determination of Cu concentration. Pasture samples were collected every 4-8 weeks for analysis of concentration of Cu and zinc (Zn). At 160 days of age, articular cartilage of long bones was examined. Gross lesions were counted and scored, then sawn and radiographed, and processed for histopathology. Lesions were given radiographic scores and histopathological scores. Maximum scores for each lesion were combined to give a total OC score for each joint and each foal. The fatness of 20 foals (10 each from Years 1 and 2) at 160 days of age was determined chemically. RESULTS: Supplementation of dams with Cu had no significant effect on the concentration of Cu in the liver of foals at birth, or on the frequency or severity of lesions in articular cartilage at 160 days of age. The Cu and Zn concentrations of pasture were similar in Years 1 and 2, and were lower than current recommendations. All foals in Year 2, and 9/10 foals in Year 1 had irregularities in cartilage that was confirmed histologically to be indicative of OC. The average number of lesions per foal was 4.7 (SD 1.1) and 5.7 (SD 1.1) in Years 1 and 2, respectively. However, the severity of the lesions was considered mild, and no foals showed any clinical evidence of OC while alive. The number of lesions in the tarsocrural (TC) joint and the TC OC score at 160 days were positively associated with average daily weight gain (ADG) in the previous 4 weeks (p=0.005 and p=0.001, respectively). There was no significant effect of sex, fatness, birthweight, weight at 160 days of age, or year of birth of the foal on the frequency and severity of OC lesions. CONCLUSIONS: Many of the lesions classified as OC, using classification systems described by other authors, were likely to be normal variations of the process of endochondral ossification. Despite the high frequency of such lesions, they were considered to be of minor significance and none were clinically evident. The distribution of lesions was not typical, and most probably reflected the subtlety of the lesions. These results support the hypothesis that Cu is an over-emphasised factor in the aetiopathogenesis of OC. The relationship between subtle macroscopic lesions and lesions resulting in clinical signs of disease requires further investigation.     

Prevalence of osteochondrosis in the limb joints of South German Coldblood horses

This study aimed to quantify the factors associated with the prevalence of the radiological signs of osteochondrosis (OC) and osteochondrosis dissecans (OCD) in South German Coldblood (SGC) horses. The prevalence of OC and OCD in fetlock and hock joints was analysed in 167 young coldblood horses with a mean age of 14 months. The presence of at least one osteochondrotic lesion (OC) in fetlock or hock joints was documented for 61.7% of the horses and 26.9% of the horses had osseus fragments. Osteochondrotic findings at the dorsal aspect of the sagittal ridge of the third metacarpal/metatarsal bone were seen in 53.9% of the horses and palmar/plantar osseous fragments in fetlock joints in 16.2% of the horses. Hock joint OC was found in 40.1% of the horses and hock OCD in 0.6%. Osteochondrotic findings in the distal part of the tibia were prevalent in 28.1% and in the lateral trochlea tali in 17.4% of the horses. The sex of the investigated horses significantly influenced the prevalence of OC in fetlock and hock joints, as well as the findings in the distal part of the tibia and lateral trochlea tali. Age at radiological examination was significant for the prevalence of OC in hock joints, palmar/plantar osseous fragments in fetlock joints and osteochondrotic findings in the distal part of the tibia. Female horses showed a 2-fold higher risk for OC in fetlock and hock joints than male horses. The distribution of the affected horses by age classes showed that radiographic signs of OC in fetlock and hock joints significantly increased at an age of about 1 year. We can conclude from our study that fetlock and hock OC is a prevalent radiographic finding in more than 1-year-old female and male SGC horses.     

Radiographic evolution of the forelimb digit in Ardenner horses from weaning to 28 months of age and its clinical significance

Ossification of collateral cartilages is a common radiographic finding in draught horses and indication of a high frequency of interphalangeal osteo-arthropathy have been found in Ardenner horses. This study first aimed at following the evolution of the digits of the forelimbs on radiographic images in a group of 32 Ardenner horses from weaning to 852 +/- 19 days and, secondly, at evaluating the clinical significance of the observed radiographic changes. At 250 +/- 21 days, only 23% of the horses presented a small separate ossification centre of collateral cartilages above the level of the distal sesamoid bone or at the level of the proximal half of the middle phalanx. At 852 +/- 19 days, 100% of the horses presented an ossification of collateral cartilages but at different stages and grades. The presence and the extent of the ossification of the lateral cartilages were not significantly associated with lameness. Phalanges changes appeared at the average age of 1 year and they were represented by entheseophytes on the dorsal surface of the middle and distal phalanges. The frequency of affected horses increased with age as well as the size of the entheseophytes. This may reflect mechanical stresses applied to the interphalangeal joints. A significant association with the presence of lameness was pointed out. The presence and the severity of phalanges changes could be of importance, in combination with a clinical examination, in improving the selection of horses able to assume work and/or leisure activities.     

The influence of sex on biochemical markers of bone turnover in dogs

Biochemical markers of bone turnover have been shown to be useful as inexpensive and noninvasive tools for monitoring skeletal health. The reference range for bone markers in dogs has been set by different age groups. However, other sources of biological variations were not fully investigated in dogs. To explore whether sex influences the interpretation of bone marker data we examined serum bone markers in 33 male and 25 female dogs. The bone markers selected for this study were: bone alkaline phosphatase (BALP) and osteocalcin (OC) as indicators of bone formation, and C-terminal telopeptide (CTx) of type I collagen as marker of bone resorption. All concentrations of bone markers were lower, but still within the reference range reported for dogs. We found statistically significant differences of the median OC and CTx serum concentrations between males and females. The results of this study suggest that there are sex differences in biochemical markers of bone turnover in dogs which should be considered in interpretation of bone marker data.     

Early changes in biomarkers of skeletal metabolism and their association to the occurrence of osteochondrosis (OC) in the horse

REASON FOR PERFORMING STUDY: Diagnosis of osteochondrosis (OC) is based on clinical signs and radiography, but alternative methods for detection at an early stage would be useful. OBJECTIVES: To determine in the juvenile horse the relationship between serum concentrations of a number of biomarkers that reflect changes in cartilage and bone turnover and age, feeding level, growth, and the occurrence of OC. METHODS: Foals were assigned to a high (n = 20) or moderate (n = 19) feeding level group from birth to age 1 year. Bodyweight, withers height and cannon width were measured. Osteoarticular status was assessed radiographically at 5.5 and 11 months in all foals, and by necropsy at 12 months for 8 foals/group. Serum biomarkers of bone (osteocalcin, CTX-1) and cartilage (CPII, C2C) metabolism were assayed at 8 time points between ages 2 and 52 weeks. Ratios between biomarkers of tissue formation and degradation were calculated at each time point. RESULTS: Consistent age-related patterns in biomarker serum concentrates were found, indicating a markedly higher metabolism before age 20 weeks but concentrations were not affected by feeding level. Bodyweight was correlated negatively to C2C and CTX-1, and withers height was positively correlated to osteocalcin and the osteocalcin/CTX-1 and CPII/ C2C ratios. Osteocalcin concentration at 2 weeks and CPII/ C2C ratio at 20 weeks had strong positive correlations to OC, as diagnosed radiographically at 5.5 months. Osteocalcin had a strong correlation with radiographically detected OC at 11 months but at that time there was no significant relationship between CPII/C2C ratio and OC. CONCLUSIONS: Occurrence of OC lesions is significantly associated with anabolic changes in bone metabolism during the first weeks post partum, given the strong relation with osteocalcin. POTENTIAL RELEVANCE: Measuring osteocalcin concentrations during the first few weeks post partum may have potential value for the prediction of risk for OC development.     

Clinical pharmacokinetics of oseltamivir and its active metabolite oseltamivir carboxylate after oral administration in horses

The aim of this study was to investigate the pharmacokinetics of oseltamivir carboxylate (OC) in horses (n=6) after oral administration of its prodrug oseltamivir. The binding rate of OC to horse plasma proteins was negligible (<1%). Oral administration of oseltamivir of 2 mg/kg body weight of oseltamivir to horses provided a plasma concentration of OC (mean maximum concentration: 257.9 ng/ml) above the inhibitory concentrations against equine influenza A viruses determined in vitro. However, because OC is rapidly eliminated from horse plasma (mean elimination half-life: 2.5 hr), administration intervals should be less than 10 hr to retain a suitable concentration when using a single dose of 2 mg/kg oseltamivir.     

Prediction of early race starts in Norwegian-Swedish Coldblooded Trotters

Background

Less than a third of Norwegian-Swedish Coldblooded Trotters (NSCTs) have started racing as three year olds since the year 2000 despite the fact that large sums are paid out as price-money in the three year season. Recruitment races are arranged by the Norwegian Trotting Association (NTA) to stimulate early training. The management of young horses varies considerably and a large majority is reared by amateurs. The aim of the present study was to identify predictors of early race starts in young NSCT horses under field conditions.

Methods

Of the 801 registered NSCT horses born in 2005, 144 were randomly selected by stratified sampling with gender and paternal progeny as stratification factors. All horses were examined clinically. Further data were collected from NTA and by interviews of breeders, owners and trainers. The set of dependent variables consisted of "passed recruitment race", "start in regular race by the end of the three year season" and "start in regular race by the end of October in the four year season". Univariate and logistic regression analyses were performed.

Results

Genetic performance potential, as indicated by best linear unbiased prediction (BLUP) indices, was the major predictor of the three dependent variables despite large variation in management. Dam''s index was a better predictor than sire''s index. However, the probability of early race starts in a horse with a low genetic performance potential can be increased by a favourable management. Examples of advantageous management factors in the present study were a flat pasture the first summer and early training. Nearly all horses racing in the three or four year seasons had passed a recruitment race in the two year season.

Conclusions

The results confirm the value of the published BLUP index as an important tool for the NSCT breeding program. Recruitment races stimulate early training.     

The selenium and vitamin E status of horses in Prince Edward Island

Serum selenium (Se), vitamin E, and resting thyroid hormone concentrations were measured in 201 horses in Prince Edward Island (PEI). Selenium concentrations were either marginal (0.0053 to 0.1200 ppm) or deficient (< 0.0053 ppm) in 79% of horses based on current reference ranges for Se in serum. Aged and young adult pleasure horses had a higher prevalence of inadequate Se concentrations compared to racehorses and broodmares (82% and 97% versus 45% and 72%, respectively). Overall, 13% of horses had inadequate (< 200 μg/dL) serum vitamin E concentrations; most of these were young pleasure horses. No horses were hypothyroid and, contrary to findings in other species, there was a positive relationship between serum thyroxine and Se concentrations (P < 0.05). We conclude that Se deficiency is widespread in PEI horse populations, especially in pleasure horses, and vitamin E deficiency is more common in young pleasure horses. Micronutrient supplementation practices employed by PEI horse owners appear inadequate to ensure sufficiency.     

Anatomic variations and degenerative changes in the collateral cartilages and middle and distal phalanges of the forelimb in Ardenner colts

The anatomic variations and the degenerative changes in the collateral cartilages and middle and distal phalanges in 6 Ardenner colts were characterized by radiography, scintigraphy, and magnetic resonance imaging (MRI). The radiographic changes were assessed between the ages of 16.5 and 25 months. An anatomic variation of the middle and distal phalanges was demonstrated in some of the colts. MRI examination of the 6 colts revealed an association between the deep digital flexor tendon cross-sectional area and body weight as well as foot circumference. Also, a thin collateral sesamoidean (suspensory navicular) ligament was observed subjectively. The presence of an extensive ossification of the collateral cartilages of the foot was found in these young horses. The ossification was characterized by the existence of 2 separate ossification centers, which had a tendency to unite. The radiographic interphalangeal degenerative lesions seen appear as bone remodeling of the dorsal edges of the middle and distal phalanx, on the insertion sites of collateral ligaments of the distal interphalangeal joint, the digital extensor tendon, and the distal interphalangeal joint capsule.     

Plasma Concentration of Insulin-like Growth Factor I (IGF-I) in Growing Ardenner Horses Suffering from Juvenile Digital Degenerative Osteoarthropathy

Degenerative osteoarthropathy resulting in a reduced active lifespan was observed in Ardenner horses. In the context of joint biology, insulin-like growth factor I (IGF-I) is a potential candidate to affect the anabolism of cartilage matrix molecules. A group of 30 Ardenner horses reared under standardized conditions from weaning were evaluated periodically from 15 to 28 months of age to detect the early manifestations of the disease. At the end of this period, horses were classified in two pathological groups related to the degree of interphalangeal degenerative osteoarthropathy based on clinical and radiographic evaluations: healthy (46.7%) and pathological (53.3%) horses. Seven sequential blood samples were taken from each horse (during the evaluation period) to study the variation of IGF-I plasma concentration. We tested the variations of the IGF-I plasma concentration during growth, and the effect of sex and of pathological classes. Significant variations were observed during the research period, with a maximum value corresponding to spring and a minimum in autumn. A significant reduction of the IGF-I plasma concentration was also observed in the pathological horses (433.5 +/- 19.5 ng/ml) compared to the healthy horses (493.9 +/- 18.2 ng/ml). An alteration in the level of this growth factor could induce a disregulation of the mechanisms involved in the local control of joint and bone tissue development.     

Articular/epiphyseal osteochondrosis in Thoroughbred foals at 5 months of age: Influences of growth of the foal and prenatal copper supplementation of the dam

AIMS: To determine the influence of copper (Cu) supplementation by injection of mares in late gestation on the frequency and severity of osteochondrosis (OC) lesions in their foals at around 160 days of age. To determine if there was any influence of the concentration of Cu in the liver, growth rate, birthweight, weight at 160 days of age, fatness, sex, or year of birth of the foal on the frequency and severity of OC lesions. To determine the influence of dam's age, and sex and birthweight of the foal on the growth rate from birth to 160 days of age, and weight at 160 days of age.

METHODS: Thirty-three Thoroughbred foals, born in two consecutive years, were weighed every 2 weeks from birth. The dams had been supplemented with parenteral Cu or saline during late gestation, and the supplementation regimens were different in each year. Foals had liver biopsies harvested at birth for determination of Cu concentration. Pasture samples were collected every 4–8 weeks for analysis of concentration of Cu and zinc (Zn). At 160 days of age, articular cartilage of long bones was examined. Gross lesions were counted and scored, then sawn and radiographed, and processed for histopathology. Lesions were given radiographic scores and histopathological scores. Maximum scores for each lesion were combined to give a total OC score for each joint and each foal. The fatness of 20 foals (10 each from Years 1 and 2) at 160 days of age was determined chemically.

RESULTS: Supplementation of dams with Cu had no significant effect on the concentration of Cu in the liver of foals at birth, or on the frequency or severity of lesions in articular cartilage at 160 days of age. The Cu and Zn concentrations of pasture were similar in Years 1 and 2, and were lower than current recommendations. All foals in Year 2, and 9/10 foals in Year 1 had irregularities in cartilage that was confirmed histologically to be indicative of OC. The average number of lesions per foal was 4.7 (SD 1.1) and 5.7 (SD 1.1) in Years 1 and 2, respectively. However, the severity of the lesions was considered mild, and no foals showed any clinical evidence of OC while alive. The number of lesions in the tarsocrural (TC) joint and the TC OC score at 160 days were positively associated with average daily weight gain (ADG) in the previous 4 weeks (p=0.005 and p=0.001, respectively). There was no significant effect of sex, fatness, birth-weight, weight at 160 days of age, or year of birth of the foal on the frequency and severity of OC lesions.

CONCLUSIONS: Many of the lesions classified as OC, using classification systems described by other authors, were likely to be normal variations of the process of endochondral ossification. Despite the high frequency of such lesions, they were considered to be of minor significance and none were clinically evident. The distribution of lesions was not typical, and most probably reflected the subtlety of the lesions. These results support the hypothesis that Cu is an over-emphasised factor in the aetiopathogenesis of OC. The relationship between subtle macroscopic lesions and lesions resulting in clinical signs of disease requires further investigation.     

Serum concentrations of keratan sulfate,osteocalcin, and pyridinoline crosslinks after oral administration of glucosamine to standardbred horses during race training

OBJECTIVE: To determine the effects of orally administered glucosamine on concentrations of markers of bone and cartilage metabolism in Standardbred horses during race training. ANIMALS: Twenty 16- to 20-month-old Standardbreds beginning race training. PROCEDURE: Horses were randomly assigned to 2 groups. One group received glucosamine hydrochloride (4 g, PO, q 12 h), and the second (control) group received glucose (4 g, PO, q 12 h). Serum samples were obtained prior to onset of the study (baseline) and at regular intervals for 48 weeks for determination of concentrations of keratan sulfate (KS), osteocalcin (OC), and pyridinoline crosslinks (PYD). RESULTS: Osteocalcin concentrations changed significantly with time; mean serum concentrations were significantly higher than baseline values for samples obtained at 24 to 48 weeks after onset of the study. Although a significant effect of time was observed for mean concentration of KS, concentrations did not differ significantly from baseline values at any time during the study when groups were analyzed separately. However, pooled analysis revealed significant increases of mean serum KS concentration at weeks 24 and 30. Significant changes in serum PYD concentrations were not detected. Oral administration of glucosamine did not significantly affect serum concentrations of any of the markers. CONCLUSIONS AND CLINICAL RELEVANCE: Increased serum OC in clinically normal Standardbreds during race training may reflect bone formation that accompanies adaptive remodeling of the appendicular skeleton. For these experimental conditions, glucosamine did not appear to exert a detectable influence on serum concentrations of these 3 markers of connective tissue metabolism.