Yoshinobu Inoue

Effect of exercise on iron metabolism in horses

We investigated the effect of exercise on iron metabolism in horses. Four horses were walked on a mechanical walker for 1 wk (pre-exercise). They then performed moderate exercise on a high-speed treadmill in the first week of the exercise and relative high in the second week and high in the third week. Serum iron was significantly lower in the third week of exercise than in the pre-exercise. Transferrin saturation (TS) was significantly lower in the first and third weeks of exercise than in the pre-exercise. Serum haptoglobin was significantly lower in the first week of exercise than in the pre-exercise and further significantly lower in the second and third weeks than in the first. The packed cell volume did not change during the experiment. The exercise significantly increased the apparent absorption of iron. Urinary iron excretion did not change throughout the experiment. Sweat iron loss did not change during the exercise. The exercise significantly increased iron balance. We considered that hemolysis is induced by moderate exercise and is further enhanced by heavy exercise, which decreases serum iron and TS. However, the increase in iron absorption compensates for the adverse effect of exercise on iron status. Therefore, exercise does not induce anemia in horses.

In vivo effects of intra-articular injection of gelatin hydrogen microspheres containing basic fibroblast growth factor on experimentally induced defects in third metacarpal bones of horses

OBJECTIVE To evaluate the effect of intra-articular injection of gelatin hydrogel microspheres containing basic fibroblast growth factor (bFGF) on experimentally induced defects in third metacarpal bones (MC3s) of horses, in vivo. ANIMALS 6 healthy adult Thoroughbreds. PROCEDURES Horses were anesthetized, and a hole (diameter, 4.5 mm) was drilled into the medial condyle of both MC3s of each horse. One milliliter (100 microg) of a solution of gelatin hydrogel microspheres (2 mg) containing bFGF was injected into the joint capsule of the right metacarpophalangeal joint of each horse (bFGF joint). One milliliter of saline (0.9% NaCl) solution was injected into the left metacarpophalangeal joint (control joint). Radiography was performed 1 day and 4, 8, 12, and 16 weeks after surgery to evaluate bone defect refilling. Sixteen weeks after surgery, multidetector-row computed tomography (MDRCT) was performed to determine the degree of refilling at the bone defect site. RESULTS Radiography revealed healing of bone defects at 4 to 12 weeks after surgery in bFGF joints and at 8 to 16 weeks after surgery in control joints. In addition, MDRCT revealed a higher degree of healing in bFGF versus control joints. Mean+/-SD MDRCT score for bFGF joints (411.7+/-135.6 Hounsfield units) was significantly higher than that for control joints (240.8+/-133.1 Hounsfield units). CONCLUSIONS AND CLINICAL RELEVANCE Treatment of horses with gelatin hydrogel microspheres that contained bFGF enhanced bone regeneration and healing of experimentally induced defects. This treatment strategy may be useful for treating horses with fractures.

Empirical Growth Curve Estimation Using Sigmoid Sub-functions that Adjust Seasonal Compensatory Growth for Male Body Weight of Thoroughbred Horses.

Thoroughbred horses are seasonal mating animals, raised in northern regions or countries. Foals born yearly in spring generally show a typical seasonal compensatory growth pattern, in which their growth rate declines in the first winter and increases in the next spring. In this study, a new empirical adjustment approach is proposed to adjust for this compensatory growth when growth curve equations are estimated, by using 1,633 male body weights of Thoroughbreds as an illustrating example. Based on general Richards growth curve equation, a new growth curve equation was developed and fit to the weight-age data. The new growth curve equation had a sigmoid sub-function that can adjust the compensatory growth, combined with the Richards biological parameter responsible for the maturity of animals. The unknown parameters included in the equations were estimated by SAS NLMIXED procedure. The goodness of fit was examined by using Akaike’s Information Criterion (AIC). The AIC values decreased from 13,053 (general Richards equation) to 12,794 (the newly developed equation), indicating the better fit of the new equation to the weight-age data. The shape of the growth curve was improved during the period of compensatory growth. The proposed method is one of the useful approaches for adjusting seasonal compensatory growth in growth curve estimations for Thoroughbreds, and for their management during the compensatory period. Based on this approach, the optimal growth curve equations can be estimated also for female body weight of Thoroughbreds or other growth traits affected by seasonal compensatory growth.

Anti-inflammatory and Intestinal Barrier-protective Activities of Commensal Lactobacilli and Bifidobacteria in Thoroughbreds: Role of Probiotics in Diarrhea Prevention in Neonatal Thoroughbreds.

ABSTRACT We previously isolated the commensal bacteria lactobacilli and bifidobacteria from the Thoroughbred intestine and prepared the horse probiotics LacFiTM, consisting of Lactobacillus ruminis KK14, L. equi KK 15, L. reuteri KK18, L. johnsonii KK21, and Bifidobacterium boum HU. Here, we found that the five LacFiTM constituent strains remarkably suppressed pro-inflammatory interleukin-17 production in mouse splenocytes stimulated with interleukin-6 and transforming growth factor-β. The protective effects of the probiotic on impaired intestinal barrier function were evaluated in Caco-2 cells treated with tumor necrosis factor-α. Evaluation of transepithelial resistance showed that all the strains exhibited intestinal barrier protective activity, with significant suppression of barrier impairment by L. reuteri KK18. The LacFiTM constituent strains were detected in neonatal LacFiTM-administered Thoroughbred feces using polymerase chain reaction denaturing gradient gel electrophoresis and culture methods. These five strains were found to be the predominant lactobacilli and bifidobacteria in the intestinal microbiota of LacFiTM-administered Thoroughbreds. Administration of LacFiTM to neonatal Thoroughbreds decreased diarrhea incidence from 75.9% in the control group (n=29 neonatal Thoroughbreds) to 30.7% in the LacFiTM-administered group (n=101 neonatal Thoroughbreds) immediately after birth to 20 weeks after birth. LacFiTM treatment also prevented diarrhea especially at and around 4 weeks and from 10 to 16 weeks. The duration of diarrhea was also shorter in the probiotics-administered group (7.4 ± 0.8 days) than in the control group (14.0 ± 3.2 days). These results indicate that the LacFiTM probiotics regulates intestinal function and contributes to diarrhea prevention.

Response of biochemical markers of bone metabolism to exercise intensity in thoroughbred horses.

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

Empirical growth curve estimation considering multiple seasonal compensatory growths of body weights in Japanese Thoroughbred colts and fillies1

Thoroughbred horses are seasonal mating animals, and their foals are born yearly in spring seasons. In northern regions or countries, the foals generally show a typical seasonal compensatory growth pattern, where their growth rate declines in winter and increases in the next spring. In this study, a new empirical approach is proposed to adjust for this compensatory growth when growth curve equations are estimated, by using BW of Japanese Thoroughbred colts and fillies raised in Hidaka, Hokkaido. Based on the traditional Richards growth curve equation, new growth curve equations were developed and fit to the weight-age data. The foals generally experience 2 major winter seasons before their debut in horseracing. The new equations had sigmoid subfunctions that can empirically adjust the first and second year compensatory growths, combined with the Richards biological parameter responsible for the maturity of animals. The unknown parameters included in the equations were estimated by SAS NLMIXED procedure. The goodness-of-fit was examined by using several indices of goodness-of-fit (i.e., Akaikes information criterion, Bayesian information criterion, -2 log likelihood, and residual sum of squares) for the multiple applications of the subfunctions. The indices indicated the best fit of the new equations including both subfunctions for the first and second compensatory growths to the weight-age data. The shapes of the growth curves were improved during the periods of compensatory growth. The proposed method is one of the useful approaches for adjusting multiple seasonal compensatory growths in growth curve estimations of Thoroughbreds and for the management of young horses during the compensatory periods.

An Approach of Estimating Individual Growth Curves for Young Thoroughbred Horses Based on Their Birthdays

ABSTRACT We propose an approach of estimating individual growth curves based on the birthday information of Japanese Thoroughbred horses, with considerations of the seasonal compensatory growth that is a typical characteristic of seasonal breeding animals. The compensatory growth patterns appear during only the winter and spring seasons in the life of growing horses, and the meeting point between winter and spring depends on the birthday of each horse. We previously developed new growth curve equations for Japanese Thoroughbreds adjusting for compensatory growth. Based on the equations, a parameter denoting the birthday information was added for the modeling of the individual growth curves for each horse by shifting the meeting points in the compensatory growth periods. A total of 5,594 and 5,680 body weight and age measurements of Thoroughbred colts and fillies, respectively, and 3,770 withers height and age measurements of both sexes were used in the analyses. The results of predicted error difference and Akaike Information Criterion showed that the individual growth curves using birthday information better fit to the body weight and withers height data than not using them. The individual growth curve for each horse would be a useful tool for the feeding managements of young Japanese Thoroughbreds in compensatory growth periods.

Empirical Percentile Growth Curves with Z-scores Considering Seasonal Compensatory Growths for Japanese Thoroughbred Horses

Percentile growth curves are often used as a clinical indicator to evaluate variations of children’s growth status. In this study, we propose empirical percentile growth curves using Z-scores adapted for Japanese Thoroughbred horses, with considerations of the seasonal compensatory growth that is a typical characteristic of seasonal breeding animals. We previously developed new growth curve equations for Japanese Thoroughbreds adjusting for compensatory growth. Individual horses and residual effects were included as random effects in the growth curve equation model and their variance components were estimated. Based on the Z-scores of the estimated variance components, empirical percentile growth curves were constructed. A total of 5,594 and 5,680 body weight and age measurements of male and female Thoroughbreds, respectively, and 3,770 withers height and age measurements were used in the analyses. The developed empirical percentile growth curves using Z-scores are computationally feasible and useful for monitoring individual growth parameters of body weight and withers height of young Thoroughbred horses, especially during compensatory growth periods.

Plasma vitamin K concentration in horses supplemented with several vitamin K homologs1

The effect of several vitamin K homologs on plasma vitamin K concentration was determined to assess their potential as a vitamin K supplement for adult horses. Sixteen Thoroughbred horses consisting of 8 mares and 8 geldings, aged 8.4 ± 3.6 yr and weighing 520.8 ± 36.1 kg, were allocated to 4 groups (n = 4). Each group was given phylloquinone, menaquinone-4, or menadione at 58 µmol/d, or no vitamin K supplement for 7 d. Plasma samples were collected before feeding, and 2, 4, and 8 h after feeding on d 7, and plasma concentrations of phylloquinone and menaquinone-4 were determined. Plasma phylloquinone concentration was greater in the phylloquinone group than in the other groups (P < 0.001). The phylloquinone concentration quadratically increased (P < 0.001) after feeding in the phylloquinone group but no changes in the plasma phylloquinone concentration were observed after feeding in the other groups. Plasma menaquinone-4 concentration was greater (P < 0.001) in the menadione group than the other groups, including the menaquinone-4 group. Menaquinone-4 concentration did not change (P = 0.192) after feeding in each group. Menaquinone-4 has been considered the most potent vitamin K homolog for bone metabolism; therefore, the present experiment indicates that menadione is a good source of vitamin K for bone health in horses because it is the only vitamin K homolog that increased the plasma concentrations of menaquinone-4.