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.     

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.     

Effect of increasing levels of undegradable intake protein on metabolic and endocrine factors in estrous cycling beef heifers

To determine the influence of three levels of undegradable intake protein (UIP) supplementation on metabolic and endocrine factors that influence reproduction, 23 yearling crossbred heifers (body condition score = 4.5 +/- 0.5; initial BW = 362 +/- 12 kg) were stratified by BW and assigned randomly to one of three supplements: 1) low UIP (1,135 g x heifer(-1) x d(-1); 30% CP, 115 g UIP, n = 7); 2) mid UIP (1,135 g x heifer(-1) x d(-1); 38% CP, 216 g UIP, n = 8); or 3) high UIP (1,135 g x heifer(-1) x d(-1); 46% CP, 321 g UIP, n = 8). Heifers were estrually synchronized before initiation of supplementation. Supplement was individually fed daily for 30 to 32 d, at which time heifers were slaughtered (d 12 to 14 of the estrous cycle) and tissues collected. Heifers were fed a basal diet of sudan grass hay (6.0% CP) ad libitum. On d 28 of supplementation (d 10 of the estrous cycle), no differences were observed (P > 0.10) in serum insulin or IGF-I among treatments. At slaughter (d 10 to 12 of the estrous cycle), treatments did not influence corpus luteum weight, cerebral spinal fluid leptin, or IGFBP; serum estradiol-17beta, progesterone, leptin, IGF-I, and IGFBP; or anterior pituitary content of IGFBP (P > 0.10). Follicular fluid IGFBP-2 and IGFBP-4 were greater in high-UIP heifers than low- or mid-UIP heifers on d 12 to 14 of the estrous cycle (P < 0.05). Basal serum LH concentrations and LH area under the curve (every 15 min for 240 min) did not differ (P > 0.10) following 28 d of supplementation (d 10 of the estrous cycle); however, basal serum FSH concentrations were greater (P = 0.06) in low- and mid- vs. high-UIP heifers (5.2 and 5.2 vs. 4.6 ng/mL, respectively), and FSH area under the curve was greater (P = 0.03) in low- vs. high-UIP heifers. At slaughter (d 12 to 14 of the estrous cycle), anterior pituitary LH and FSH content and steady-state mRNA encoding alpha, LHbeta, and GnRH receptor did not differ (P > 0.10) among treatments. However, FSHbeta mRNA was increased approximately twofold (P = 0.03) in mid vs. low UIP. In summary, low and mid levels of UIP supplements fed to estrous cycling beef heifers seemed to enhance pituitary expression and/or secretion of FSH relative to high levels of UIP. Moreover, high-UIP supplementation was associated with increased low-molecular-weight IGFBP compared with supplementation of low and mid levels of UIP. These data suggest that differing levels of UIP supplementation may alter pituitary and ovarian function, thereby influencing reproductive performance in beef heifers.     

Measurement of synovial fluid and serum concentrations of the 846 epitope of chondroitin sulfate and of carboxy propeptides of type II procollagen for diagnosis of osteochondral fragmentation in horses

OBJECTIVE: To determine whether serum or synovial fluid concentrations of chondroitin sulfate epitope 846 and carboxy propeptides of type II collagen (CPII) can be used to diagnose osteochondral fragmentation (OC) in horses. ANIMALS: 38 horses with unilateral OC of the radiocarpal (n = 31) or intercarpal (33) joints and 8 clinically and radiographically normal horses. Procedures-For horses with OC, serum and synovial fluid concentrations of epitope 846, CPII, and keratan sulfate (KS) were determined, along with synovial fluid WBC counts and total protein concentrations. Serum epitope 846, CPII, and KS concentrations were measured in control horses. RESULTS: Synovial fluid epitope 846 and total protein concentrations were significantly higher in the joints with OC than in unaffected joints, but CPII and KS concentrations and WBC counts were not. Synovial fluid total protein and 846 epitope concentrations were linearly related to grade of OC. Serum epitope 846 and CPII concentrations were significantly higher in horses with OC than in control horses. Discriminant analysis allowed 27 of 34 (79%) horses to be correctly classified as having or not having OC on the basis of serum epitope 846 and CPII concentrations. CONCLUSIONS: Results suggest that serum and synovial fluid concentrations of epitope 846 and CPII are associated with OC. Increases in concentrations of epitope 846 and CPII suggest that increased synthesis of cartilage aggrecan and type II procollagen may be associated with OC. CLINICAL RELEVANCE: Measurement of serum epitope 846 and CPII concentrations may be useful in the diagnosis of OC in horses.     

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.     

Association of maximum voluntary dietary intake of freeze-dried garlic with Heinz body anemia in horses

OBJECTIVE: To characterize hematologic and clinical consequences of chronic dietary consumption of freeze-dried garlic at maximum voluntary intake in horses. ANIMALS: 4 healthy sex- and age-matched horses. PROCEDURE: An initial garlic dose (0.05 g/kg, twice daily) was fed to 2 horses in a molasses carrier as part of their normal ration and was gradually increased to maximum voluntary intake (0.25 g/kg, twice daily) over 41 days. Dietary supplementation then continued for a total of 71 days. Two control horses were fed molasses with no garlic with their ration. Blood samples were collected weekly and analyzed for hematologic and biochemical changes, including the presence of Heinz bodies. Recovery of affected blood values was followed for 5 weeks after termination of dietary supplementation with garlic. RESULTS: At a daily dose of > 0.2 g/kg, horses fed garlic developed hematologic and biochemical indications of Heinz body anemia, as characterized by increases in Heinz body score (HBS), mean corpuscular volume (MCV), mean corpuscular hemoglobin, platelet count, and serum unconjugated and total bilirubin concentrations and decreases in RBC count, blood hemoglobin concentration, mean corpuscular hemoglobin concentration, and serum haptoglobin concentration. Recovery from anemia was largely complete within 5 weeks after termination of dietary supplementation with garlic. Heinz body score and MCV remained high at the end of the 5-week recovery period. CONCLUSIONS AND CLINICAL RELEVANCE: Horses will voluntarily consume sufficient quantities of garlic to cause Heinz body anemia. The potential for garlic toxicosis exists when horses are chronically fed garlic. Further study is required to determine the safe dietary dose of garlic in horses.     

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.     

Chlortetracycline supplementation of yearling dairy heifers

Chlortetracycline is an antibiotic that is used to increase weight gain, efficiency of gain, carcass grade, and conception rates. The objective of this experiment was to evaluate the effects of supplementation of 350 mg/d of chlortetracycline on ADG, G:F, BCS, thyroxine, and systemic glucose concentrations in yearling dairy heifers. Forty 12-mo-old Holstein heifers (initial BW = 363 +/- 21 kg) were housed in a free-stall barn with ad libitum access to feed and water for 104 d. A transition period was begun 14 d before the age of 12 mo to acclimate the heifers to the diet. The chlortetracycline-fed group (n = 20) consumed 328 +/- 8.2 mg of chlortetracycline/heifer daily. Measurements for BW, withers and hip heights, BCS, and health score were recorded weekly. Dry matter intake was measured daily. Blood was sampled every 4 d to determine plasma thyroxine and glucose concentrations and every 2 d to determine progesterone concentrations. Heifers were artificially inseminated on the first observed standing heat after 13 mo of age. There were no effects of chlortetracycline on ADG, G:F, withers and hip heights, BCS, blood glucose concentrations, peak progesterone concentrations, health, or conception rate. There was an interaction between treatment and time for chlortetracycline on serum thyroxine concentration. In the beginning of the experiment, serum thyroxine concentration was lower in heifers supplemented with chlortetracycline. There was no difference between treatments in thyroxine concentration at the end of the experiment. Chlortetracycline supplementation was not beneficial for yearling dairy heifers.     

Digestible energy intake, dry matter digestibility and mineral status of grazing New Zealand Thoroughbred yearlings

AIM: To measure the nutritive value of pasture in terms of digestible energy intake (DEI) and dry matter (DM) digestibility, and to determine the effect of Ca, P, Cu, Zn, and Se supplementation on growth rate and degree of physeal swelling in Thoroughbred yearlings grazed on pasture. METHODS: Fourteen yearling horses were randomly allocated to 1 of 2 groups and rotationally grazed on a ryegrass/white clover pasture for 7 months. One group was supplemented daily with a mineral mix. Liveweight changes were recorded at fortnightly intervals and pasture mineral composition determined at monthly intervals. The DM intake (DMI) was determined from daily faecal DM outputs divided by the indigestible DM fraction (1 - digestible DM) determined from a digestibility study. The DEI was determined from the difference between the gross energy intake and the gross energy faecal output. The DM, gross energy content, crude protein (CP), soluble carbohydrate, acid detergent fibre (ADF), neutral detergent fibre (NDF), lipid and mineral composition of pasture offered and faeces were analysed and their digestibility or apparent absorption calculated. RESULTS: The DM digestibility of pasture averaged 0.64 while the daily DMI and DEI of a 350 kg yearling, gaining 0.6 kg liveweight/day, averaged 6.9 kg DM/day and 78 MJ DE/day, respectively. The mineral supplement had no significant effect on the growth of yearlings grazing pasture that had a mineral composition of (g/kg DM) Na 0.17, K 32.8, Ca 3.6, P 3.4, Mg 2.1, S 3.0, and (mg/kg DM) Cu 7.9, Fe 177, Mn 83, Zn 28 and Se 0.016. Plasma Se was elevated (e.g. 40-218 microg/l) by Se supplementation while Cu supplementation had no effect on plasma and liver Cu concentrations. Physeal swelling was observed in all horses, regardless of treatment. CONCLUSION: Good quality ryegrass/white clover pasture that had a DE content of 11.3 MJ/kg DM and adequate mineral composition (as observed in this study, with the possible exception of Se) will provide an adequate intake of nutrients to ensure good growth in yearling 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 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.     

Dietary supplementation of a Saccharomyces cerevisiae fermentation product attenuates exercise-induced stress markers in young horses

Mitigation of exercise-induced stress is of key interest in determining ways to optimize performance horse health. To test the hypothesis that dietary supplementation of a Saccharomyces cerevisiae fermentation product would decrease markers of exercise-induced stress and inflammation in young horses, Quarter Horse yearlings (mean ± SD; 9 ± 1 mo) were randomly assigned to receive either no supplementation (CON; n = 8) or 21 g/d S. cerevisiae fermentation product (10.5 g/feeding twice daily; SCFP; n = 10) top-dressed on a basal diet of custom-formulated grain as well as ad libitum Coastal bermudagrass hay. After 8 wk of dietary treatments, horses underwent a 2-h submaximal exercise test (SET) on a free-stall mechanical exerciser. Serum was collected before dietary treatment supplementation (week 0), at week 8 pre-SET, and 0, 1, and 6 h post-SET and analyzed for concentrations of cortisol and serum amyloid A (SAA) by commercial enzyme-linked immunosorbent assay (ELISA) and for cytokine concentrations by commercial bead-based ELISA. Data were analyzed using linear models with repeated measures in SAS v9.4. From week 0 to 8 (pre-SET), serum cortisol decreased (P = 0.01) and SAA did not change, but neither were affected by diet. Serum concentrations of all cytokines decreased from week 0 to 8 (P ≤ 0.008), but granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor, and interleukin-8 (IL-8) decreased to a greater extent in CON than in SCFP horses (P ≤0.003). In response to the week 8 SET, serum cortisol increased in all horses (P < 0.0001) but returned to pre-SET levels by 1 h post-SET in horses receiving SCFP. At 6 h post-SET, cortisol concentrations in CON horses returned to pre-SET concentrations, whereas cortisol declined further in SCFP horses to below pre-SET levels (P = 0.0002) and lower than CON (P = 0.003) at that time point. SAA increased at 6 h post-SET in CON (P < 0.0001) but was unchanged through 6 h in SCFP horses. All cytokines except G-CSF increased in response to the SET (P < 0.0001) but showed differing response patterns. Concentrations of IL-1β, IL-6, and tumor necrosis factor-alpha were lesser (P ≤ 0.05), and concentrations of G-CSF and IL-18 tended to be lesser (P ≤ 0.09) in SCFP compared with CON horses throughout recovery from the SET. In summary, 8 wk of dietary supplementation with 21 g/d of SCFP may mitigate cellular stress following a single, prolonged submaximal exercise bout in young horses.     

Effects of oral administration of levothyroxine sodium on serum concentrations of thyroid gland hormones and responses to injections of thyrotropin-releasing hormone in healthy adult mares

OBJECTIVE: To determine the effects of levothyroxine sodium (L-T4) on serum concentrations of thyroid gland hormones and responses to injections of thyrotropin-releasing hormone (TRH) in euthyroid horses. ANIMALS: 12 healthy adult mares. PROCEDURE: 8 horses received an incrementally increasing dosage of L-T4 (24, 48, 72, or 96 mg of L-T4/d) for weeks 1 to 8. Each dose was provided for 2 weeks. Four additional horses remained untreated. Serum concentrations of total triiodothyronine (tT3), total thyroxine (tT4), free T3 (fT3), free T4 (fT4), and thyroid-stimulating hormone (TSH) were measured in samples obtained at weeks 0, 2, 4, 6, and 8; 1.2 mg of TRH was then administered i.v., and serum concentrations of thyroid gland hormones were measured 2 and 4 hours after injection. Serum reverseT3 (rT3) concentration was also measured in the samples collected at weeks 0 and 8. RESULTS: Treated horses lost a significant amount of weight (median, 19 kg). Significant treatment-by-time effects were detected for serum tT3, tT4, fT3, fT4, and TSH concentrations, and serum tT4 concentrations were positively correlated (r, 0.95) with time (and therefore dosage) in treated horses. Mean +/- SD serum rT3 concentration significantly increased in treated horses (3.06 +/- 0.51 nmol/L for week 8 vs 0.74 +/- 0.22 nmol/L for week 0). Serum tT3, tT4, fT3, and TSH concentrations in response to TRH injections differed significantly between treated and untreated horses. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of levothyroxine sodium increased serum tT4 concentrations and blunted responses toTRH injection in healthy euthyroid horses.     

Dietary fish oil supplementation affects serum fatty acid concentrations in horses

Thirteen horses of Thoroughbred or Standardbred breeding were used to study the effect of dietary fish oil supplementation on blood lipid characteristics. Horses were assigned to either fish oil (n = 7) or corn oil (n = 6) treatment groups for 63 d. The fish oil contained 10.8% eicosapentaenoic acid (EPA) and 8% docosahexaenoic acid (DHA). Each horse received timothy hay and a mixed-grain concentrate at rates necessary to maintain BW. Oil (corn or fish) was top-dressed on the concentrate daily at a rate of 324 mg/ kg of BW. The n-6:n-3 ratio was approximately 3.6:1 for horses receiving the corn oil diet and 1.4:1 for horses receiving the fish oil diet. Horses were exercised 5 d/wk during the study. Before supplementation, there was no difference in the concentrations of any serum fatty acids between the 2 treatment groups. The mean basal concentrations of EPA and DHA on d 0 were 0.04 and 0.01 mg/mL, respectively. After 63 d, horses receiving the fish oil treatment, but not those receiving the corn oil treatment, had increased concentrations of EPA and DHA (P <0.05). Fish oil supplementation for 63 d also increased the concentrations of C22:0, C22:1, and C22:5 fatty acids (P <0.05). Overall, horses receiving fish oil had a decreased concentration of n-6 fatty acids (P <0.05) and a greater concentration of n-3 fatty acids (P <0.01), resulting in a lower n-6:n-3 fatty acid ratio after 63 d (P <0.05). Serum cholesterol concentrations increased (P <0.05) during the supplementation period in horses receiving the corn oil but not in horses receiving the fish oil. Compared with horses receiving corn oil, horses receiving fish oil had lower serum triglycerides at d 63 (P <0.05). These results demonstrate that 63 d of fish oil supplementation at 324 mg/kg of BW was sufficient to alter the fatty acid profile and blood lipid properties of horses receiving regular exercise.     

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.     

Dietary omega-3 fatty acid supplementation does not impair vitamin E status or promote lipid peroxidation in growing horses

Omega-3 (n-3; ω-3) fatty acids (FA) are often included in the diet for their potential health benefits. However, because oxidative potential is increased with the degree of unsaturation in vitro, polyunsaturated FA such as eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3) may be at increased risk of lipid peroxidation. We aimed to determine the effects of dietary n-3 FA supplementation on antioxidant status and lipid peroxidation in yearling horses. Quarter Horses (mean ± SEM; 14.6 ± 0.2 mo) were randomly assigned to receive no n-3 FA supplementation (CON; n = 6) or 60 mg n-3/kg body weight from milled flaxseed (FLAX; n = 6) or encapsulated fish oil (FISH; n = 6). All horses received a basal diet of mixed grain concentrate fed individually at 1.5% body weight (dry matter basis) and ad libitum bahiagrass pasture forage. Blood samples were obtained before and after 70 d of supplementation to evaluate vitamin E, selenium, lipids, antioxidant status, and oxidative stress. Data were analyzed using a mixed model ANOVA with repeated measures. Supplementation with n-3 FA did not reduce serum vitamin E or Se and, in fact, elevated (P ≤ 0.0003) vitamin E status in FISH horses. At day 70, serum triglycerides were lower in FISH and FLAX horses than CON horses (P ≤ 0.02) and F2-isoprostanes were lower in FISH than CON horses (P = 0.0002). Dietary n-3 FA had no effect on cholesterol, reduced and oxidized glutathione, glutathione peroxidase, and thiobarbituric acid-reactive substances. In growing horses fed to meet their vitamin E requirements, supplementation with 60 mg n-3/kg body weight did not negatively affect vitamin E status or promote lipid peroxidation. Elevated vitamin E status in horses fed FISH, coupled with lower serum F2-isoprostanes, further suggest that the longer-chain, highly unsaturated n-3 FA, EPA and DHA, may actually attenuate lipid peroxidation.     

Plasma and serum concentrations of phenylbutazone and oxyphenbutazone in racing Thoroughbreds 24 hours after treatment with various dosage regimens

The plasma and serum concentrations of phenylbutazone (PBZ) and oxyphenbutazone were measured in 158 Thoroughbred horses after various doses of PBZ wer given. All horses were competing or training at racetracks in various parts of the country. All horses used in the study had not been given PBZ 24 hours before they were placed on a specific dosage schedule. Samples were collected 24 hours after the last PBZ administration. Four grams of PBZ were given daily by stomach tube, paste, or tablet for 3 days. On day 4, 24 hours before sample collection, an IV dose of 2 g of PBZ was given, regardless of the dose and method of administration. The 24-hour PBZ plasma concentrations were 3.51, 6.13, and 6.40 micrograms/ml, respectively. After 2 g of PBZ was administered IV daily for 4 days, the plasma PBZ concentration was 4.16 g/ml; after a single 2-g IV administration, the serum concentration was 0.87 g/ml. Concentrations of oxyphenbutazone were 3.35 (stomach tube), 4.29 (paste), 3.60 (tablet), 3.65 (4-day IV), and 1.11 g/ml (single IV). A significant relationship was not found between the serum and the urinary concentrations at this 24-hour measurement. Split samples sent to various laboratories confirmed the stability of high-performance liquid chromatography as a method of analysis.     

Biochemical analysis of normal articular cartilage in horses

Articular cartilage specimens from the distal articular surface of 32 radiocarpal bones from 24 2- to 5-year-old horses were analyzed. The total collagen content was determined on the basis of the 4-hydroxyproline content, using a colorimetric method. A method for estimating the proportions of types-I and -II collagen by measuring spectrophotometric densities of specific cyanogen bromide peptide bands from mixtures of types-I and -II collagen on sodium dodecyl sulfate-polyacrylamide gels was used. The cyanogen bromide peptides representative of each collagen types-I and -II were identified. The peptide ratios were then computed for each of several standards of type-I and -II mixtures. A standard curve was derived from the correlation between these ratios and the corresponding proportions of type-II collagen in standard mixtures. Galactosamine and glucosamine content (hexosamines) were measured by ion chromatography. The galactosamine-to-glucosamine ratio, chondroitin sulfate and keratan sulfate values, and total glycosaminoglycan content were derived from the measured hexosamine content. The total collagen content averaged 556 mg/g (55.6 mg/100 mg) of tissue (dry weight, [dw]). Type-II collagen was the major collagen type in normal articular cartilage specimens. The ratio of the area under the alpha 1 (II)CB10 peak to the area under the alpha 1 (I)CB 7,8 + alpha 1 (II)CB11 peak was a second-order polynomial function of the proportion of type-II collagen in the specimens. The mean galactosamine and glucosamine content were 20.6 mg/g and 7.9 mg/g (dw), respectively. The mean galactosamine-to-glucosamine ratio was 3.74 +/- 0.62.(ABSTRACT TRUNCATED AT 250 WORDS)     

Meal size and feeding frequency influence serum leptin concentration in yearling horses

Energy is an essential nutrient for all horses, and it is especially important in performance horses, pregnant and lactating mares, and young growing horses. A negative energy balance in horses such as these may result in unsatisfactory performance, decreased fertility, or slow growth. Therefore, ensuring adequate energy intake is an important aspect of the nutritional management of the equine. This study was undertaken to investigate the effects of feeding large, carbohydrate-rich, concentrate meals on the satiety-inducing hormone, leptin. Three groups of yearling horses were rotated through 3 feeding schedules in a replicated 3x3 Latin square design. Horses were fed 2, 3, or 4 times per day (2x, 3x, and 4xfeeding schedules, respectively), each for a period of 11 d, with the total amount of daily feed held constant. Horses were weighed and BCS was determined on the first day of each period. Blood samples were collected before the morning meal on d 1, 4, and 7 of each period. Additionally, blood was sampled for the last 24 h of the 2xand 4xdietary periods. Neither weight nor BCS changed during the study (P = 0.99 and P = 0.28, respectively). Both mean and peak plasma glucose were greatest in 2xhorses (P < 0.05), as were mean areas under the curve. Serum leptin concentration increased in 2xhorses (P < 0.05), but not in horses fed 3 or 4 times daily. Leptin was elevated in horses with greater BCS (P < 0.05) and increased steadily throughout the study (P < 0.05). Data from the 24-h collection indicated that 2xhorses had fluctuations in leptin production throughout the day (P < 0.05), whereas horses fed 4 times daily did not. Overall, this study indicates that feeding horses 2 large concentrate meals daily can increase mean serum leptin concentrations and may cause fluctuations in leptin production over a 24-h period. This departure from baseline leptin concentration has the potential to affect appetite, along with numerous other physiological processes.     

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.