Influence of Exercise and Intra-articular Site on Canals in Articular Calcified Cartilage of Equine Third Carpal Bones.

The third carpal bone (C3) responds to exercise by adaptive modeling of bone and articular calcified cartilage along the dorsal load path. Canals penetrating articular calcified cartilage, thought to contain vascular tissue, are reported in numerous species. Their significance remains unclear. Our objective was to determine if the number of canals was significantly different in strenuously exercised and control young horses and in a site of intermittent high loading compared to sites sustaining lower habitual loads. Volumetric bone mineral density in the radial facet of C3 of strenuously exercised and gently exercised (control) 19-month-old thoroughbred horses (n= 6/group) was determined by peripheral quantitative computed tomography. The hyaline cartilage was corroded to expose the surface of articular calcified cartilage. The number of canals penetrating the articular calcified cartilage surface in en face scanning electron microscopy images was compared in 4 regions. Volumetric bone mineral density of C3 was significantly greater (P= .004) in strenuously exercised horses. There were 2 morphologically distinct groups of canals and significantly fewer (P= .006) large canals in the dorsal than in the palmar aspect of C3 in control but not in exercised horses. Roughly circular depressions in the articular calcified cartilage surface around apparently forming canals were visible in some samples and have not been previously described in the literature. The canals may be evidence of chondroclastic activity reaching the interface of hyaline and calcified cartilage. Further work is needed to elucidate the relationships between presence of canals and the responses to exercise and to joint disease.

Voluntary exercise in pregnant rats improves post-lactation maternal bone parameters but does not affect offspring outcomes in early life.

OBJECTIVES: The objectives of this study were to examine the effects of voluntary exercise during pregnancy on maternal post-lactation bone parameters and offspring growth. METHODS: Pregnant Wistar rats were housed in conventional cages (control), or were housed in raised cages requiring them to rise to an erect, bipedal stance to obtain food/water, throughout pregnancy. Dual energy X-ray absorptiometry and peripheral quantitative computed tomography scans were performed pre-mating and post-weaning. Maternal stress was assessed by fecal corticosterone measurement. Offspring weights were assessed at postnatal days 1 and 25 (weaning). RESULTS: Changes in bone mineral over the pregnancy/lactation period were site-specific. Exercise did not affect loss of bone mineral from the lumbar spine, but did attenuate the loss of trabecular bone mineral from the tibial metaphysis and enhance the strength strain index and cross-sectional moment of inertia at the tibial diaphysis (P≤0.05) in dams in the exercised group. Fecal corticosterone did not differ between dam groups. There were no significant differences in offspring weight between the exercised and control group at either time point. CONCLUSIONS: Voluntary exercise in the pregnant rat can improve some post-lactation bone parameters and does not adversely affect early postnatal outcomes of the offspring.

Combined nanoindentation testing and scanning electron microscopy of bone and articular calcified cartilage in an equine fracture predilection site.

Condylar fracture of the third metacarpal bone (Mc3) is the commonest cause of racetrack fatality in Thoroughbred horses. Linear defects involving hyaline articular cartilage, articular calcified cartilage (ACC) and subchondral bone (SCB) have been associated with the fracture initiation site, which lies in the sagittal grooves of the Mc3 condyle. We discovered areas of thickened and abnormally-mineralised ACC in the sagittal grooves of several normal 18-month-old horses, at the same site that linear defects and condylar fracture occur in older Thoroughbreds and questioned whether this tissue had altered mechanical properties. We embedded bone slices in PMMA, prepared flat surfaces normal to the articular surface and studied ACC and SCB using combined quantitative backscattered electron scanning electron microscopy (qBSE) and nanoindentation testing: this allowed correlation of mineralisation density and tissue stiffness (E) at the micron scale. We studied both normal and affected grooves, and also normal condylar regions. Large arrays of indentations could be visualised as 2-dimensional maps of E with a limit to resolution of indentation spacing, which is much larger than qBSE pixel spacing. ACC was more highly mineralised but less stiff in early linear defects than in control regions, while subchondral bone was more highly mineralised and stiffer in specimens with early linear defects than those without. Thus both ACC and SCB mineralisation may be abnormal in a class of early linear defect in 18-month-old Thoroughbred horses, and this may possibly contribute to later fracture of the Mc3 condyle.

Assessment of bone response to conditioning exercise in the radius and tibia of young thoroughbred horses using pQCT.

OBJECTIVES: To assess the effect of conditioning exercise on bone parameters at multiple sites in the radius and tibia of young Thoroughbred horses. METHODS: The left and right radius and tibia were obtained from twelve horses, six of which had received conditioning exercise and six which formed the control group. Each bone was scanned at 5% intervals along its entire length using pQCT. RESULTS: Bone strength, bone area and periosteal circumference were significantly greater for the group of conditioned horses in both the radius and tibia. Volumetric bone mineral density was lower while bone mineral content, endocortical circumference and polar moment of inertia were higher in the conditioned group of horses but the significance of these differences varied between the two bones. Cortical thickness was not significantly different between the groups in either bone. CONCLUSIONS: Conditioning exercise stimulated a significant increase in the strength of both bones that could be attributed mainly to an increase in bone size, rather than differences in bone mineral content or density. The radius and tibia exhibited differences in the significance of changes in several bone parameters suggesting that not all bones respond in an identical fashion to imposed exercise.

Radiographic characterisation of spinal curvature development in farmed New Zealand Chinook salmon Oncorhynchus tshawytscha throughout seawater production.

Spinal anomalies are a recognised source of downgrading in finfish aquaculture, but identifying their cause(s) is difficult and often requires extensive knowledge of the underlying pathology. Late-onset spinal curvatures (lordosis, kyphosis, scoliosis) can affect up to 40% of farmed New Zealand Chinook (king) salmon (Oncorhynchus tshawytscha) at harvest, but little is known about their pathogenesis. Curvature development was radiographically documented in two related cohorts of commercially-farmed Chinook salmon throughout seawater production to determine (1) the timing of radiographic onset and relationships between (2) the curvature types, (3) the spinal regions in which they develop and (4) their associations with co-existing vertebral body anomalies (vertebral compression, fusion and vertical shift). Onset of curvature varied between individuals, but initially occurred eight months post-seawater transfer. There were strong associations between the three curvature types and the four recognised spinal regions: lordosis was predominantly observed in regions (R)1 and R3, kyphosis in R2 and R4, manifesting as a distinct pattern of alternating lordosis and kyphosis from head to tail. This was subsequently accompanied by scoliosis, which primarily manifested in spinal regions R2 and R3, where most of the anaerobic musculature is concentrated. Co-existing vertebral body anomalies, of which vertebral compression and vertical shift were most common, appeared to arise either independent of curvature development or as secondary effects. Our results suggest that spinal curvature in farmed New Zealand Chinook salmon constitutes a late-onset, rapidly-developing lordosis-kyphosis-scoliosis (LKS) curvature complex with a possible neuromuscular origin.

Influence of exercise and joint topography on depth-related spatial distribution of proteoglycan and collagen content in immature equine articular cartilage.

REASONS FOR PERFORMING STUDY: There is ample evidence on topographical heterogeneity of the principal biochemical components of articular cartilage over the surface of the joint and the influence of loading thereon, but no information on depth-related zonal variation in horses. OBJECTIVES: To study depth-related zonal variation in proteoglycan (PG) and collagen content in equine articular cartilage. METHODS: Two techniques (safranin-O densitometry and Fourier transform infrared spectroscopy) were applied to sections of articular cartilage from the proximal phalangeal bone of the metacarpophalangeal joint of 18-month-old Thoroughbreds that had been raised at pasture from age 0-18 months without (PASTEX) and with (CONDEX) additional exercise. Two sites were investigated: site 1 at the joint margin that is unloaded at rest or at slow gaits, but subjected to high-intensity loading during athletic activity; and site 2, a continuously, but less intensively, loaded site in the centre of the joint. RESULTS: Proteoglycan values increased from the surface to the deep layers of the cartilage, collagen content showed a reverse pattern. PG content was significantly higher at site 2 in both PASTEX and CONDEX animals without an effect of exercise. In the PASTEX animals collagen content was significantly higher at site 1, but in the CONDEX group the situation was reversed, due to a significant exercise effect on site 1, leading to a reduced collagen content. CONCLUSIONS: Collagen and PG content gradients agree with findings in other species. The observations on PG levels suggest that the exercise level was not strenuous. The collagen results in the PASTEX group confirmed earlier findings, the lower levels at site 1 in the CONDEX group being possibly due to an advancement of the physiological maturation process of collagen remodelling. POTENTIAL RELEVANCE: This study confirms earlier observations that even moderate variations in exercise level in early age may have significant effects on the collagen network of articular cartilage.

Influence of intensity and changes of physical activity on bone mineral density of immature equine subchondral bone.

REASONS FOR PERFORMING STUDY: Subchondral bone provides structural support to overlying articular cartilage and plays an important biomechanical role in osteochondral diseases. Mechanical features of bone correlate strongly with bone mineral density, which is directed by the loading conditions to which the tissue is subjected. OBJECTIVE: To investigate the influence of physical activity levels on subchondral bone mineral density (sBMD) in foals during early development. METHODS: Three groups of foals were subjected to different physical activity levels from birth until age 5 months. A proportion of these foals were subjected to euthanasia at 5 months while remaining foals were subjected to similar physical activity levels for 6 months until euthanasia at 11 months. Osteochondral specimens were collected for measurement of sBMD with peripheral quantitative computed tomography at 2 differently loaded anatomical sites of the proximal phalangeal bone at 1, 2, 3, 4 and 5 mm depth from the osteochondral junction. RESULTS: Growth significantly increased sBMD but by a different amount depending on anatomical location and physical activity level. Significantly higher sBMD was found at the habitually loaded central area in comparison to the intermittently peak loaded marginal site. Exercise increased sBMD throughout the whole depth of analysed tissue, but changes were generally more obvious at a depth of 2 mm. Interestingly, foals subjected to additional sprint training preserved the exercise-induced sBMD increase at the habitually loaded central area during the 6 months of the second phase of the study. CONCLUSIONS: Habitual low-intensity loading elicits a greater response in sBMD in quantitative terms than high-intensity low-frequency loading at the sites investigated in this study. Future sBMD may be influenced by means of well-tailored exercise regimens at young age. POTENTIAL RELEVANCE: Specific physical activity levels during early development may potentially reduce the prevalence of osteochondral injury later in life.

Exercise does not affect stiffness and mineralisation of third metacarpal condylar subarticular calcified tissues in 2 year old thoroughbred racehorses.

Impact exercise has a profound effect in increasing volumetric density of epiphyseal bone, as clearly shown in 2 year old thoroughbred racehorses from which we derived the tissue studied in the present investigation. Here, we asked the question whether the fabric-level properties of the mineralised tissues immediately below hyaline articular cartilage which transmit the extra loads are themselves altered in consequence. We therefore studied the nanoindentation elastic modulus and its relationship to the concentration of mineral determined by quantitative backscattered electron imaging in the heavily loaded palmar medial and lateral condyles of the distal third metacarpal bone (Mc3) of 4 untrained and 4 trained 2-year old Thoroughbred racehorses. We found no difference between trained and untrained horses in either subchondral bone or calcified cartilage in the mean stiffness or mineral content or their correlation. Thus neither articular calcified cartilage nor the immediately adjacent subchondral bone were affected by exercise, even though they transmitted the higher load associated with athletic training through to the deeper bone, which itself responded floridly to exercise. Under the circumstances of this experiment and at least in the very small regions studied, therefore, the structure of these two tissues was apparently optimised to function.

The acute effect of vibration exercise on concentric muscular characteristics.

This study was designed to compare the acute effect of vibration exercise with a concentric-only activity (arm cranking) on concentric-only muscle action using an upper body isoinertial exercise. Twelve healthy, physically active men, 30.0y+/-6.1 (mean+/-S.D.); height 1.81m+/-0.06; and weight 83.4kg+/-9.7, performed four maximal prone bench pull (PBP) efforts before and after a 5-min period of three different interventions: (1) acute vibration exercise (VBX); (2) arm cranking (AC); and (3) control (no exercise) (NVBX). Electromyography (EMG) activity was assessed from the middle trapezius muscle during PBP. Acute VBX was induced with an electric-powered dumbbell (DB) (frequency 26Hz, amplitude 3mm), with 30-s exposures at five different shoulder positions. NVXB was performed with the participants holding the DB with the machine turned off, and AC was performed at 25W. There was a significant (intervention x pre-post) interaction such that acute VBX and AC enhanced peak power by 4.8% (p<0.001) and 3.0% (p<0.001), respectively, compared to NVBX (-2.7%). However, there was no effect of any treatments on EMG activity compared to the control. In conclusion, acute VBX provides an acute ergogenic effect which potentiates concentric-only muscle performance, though not to a significantly greater extent than concentric (arm cranking) exercise.

Effects of exercise on tenocyte cellularity and tenocyte nuclear morphology in immature and mature equine digital tendons.

REASON FOR PERFORMING STUDY: The injury-prone, energy-storing equine superficial digital flexor tendon (SDFT) of the mature performance horse has a limited ability to respond to exercise in contrast with the noninjury-prone, anatomically opposing common digital extensor tendon (CDET). Previous studies have indicated low levels of cellular activity in the mature SDFT, but in foal tendons the tenocytes may still have the ability to adapt positively to increased exercise. OBJECTIVES: To measure tenocyte densities and types in histological sections from the SDFT and CDET of horses from controlled long-term, short-term and foal exercise studies. METHODS: Specimens were collected from mid-metacarpal segments of the CDET and SDFT for each horse and processed for histology; central and peripheral regions of the SDFT cross-section were analysed separately (SDFTc, SDFTp). Tenocyte nuclei were counted in a total area of 1.59 mm(2) for each tendon region in each horse. Each nucleus was classified as type 1 (elongate and thin), type 2 (ovoid and plump) or type 3 (chondrocyte-like); type 1 cells are proposed to be less synthetically active than type 2 cells. RESULTS: No significant differences were noted between exercise and control groups in any of the studies, with the exception of an exercise-related reduction in the proportion of type 1 tenocytes for all tendons combined in the long-term study. There were tendon- and site-specific differences in tenocyte densities and proportions of type 1 and 2 cells in all 3 studies. CONCLUSIONS AND POTENTIAL RELEVANCE: There was no indication that exercise increased tenocyte density or proportions of the (theoretically) more active type 2 cells in immature horses (short-term and foal studies), perhaps because the training regimens did not achieve certain threshold strain levels. In the foal study these findings can still be interpreted positively as evidence that the training regimen did not induce subclinical damage.

The influence of exercise during growth on ultrasonographic parameters of the superficial digital flexor tendon of young Thoroughbred horses.

REASON FOR PERFORMING STUDY: Conditioning by early training may influence the composition of certain musculoskeletal tissues, but very few data exist on its effect during growth on tendon structure and function. OBJECTIVES: To investigate whether conditioning exercise in young foals would lead to any ultrasonographically detectable damage to the superficial digital flexor tendon or an increase in cross-sectional area (CSA). METHODS: Thirty-three Thoroughbred foals reared at pasture were allocated to 2 groups: control (PASTEX) allowed exercise freely at pasture; and CONDEX, also at pasture, began conditioning exercise from mean age 21 days over 1030 m on a purpose-built oval grass track, for 5 days/week until mean age 18 months. Foals were observed daily, and underwent orthopaedic examination monthly. Ultrasonographic images of the superficial digital flexor tendon (SDFT) at the mid-metacarpal level of both forelimbs were obtained in all foals at ages 5, 8, 12, 15 and 18 months. CSA was validated (r(2) = 0.89) by determining CSA from digital photographs of the transected SDFT surface from 12 of the horses necropsied at age 17.1 months. RESULTS: here was no clinical or ultrasonographic evidence of tendonopathy in either group and the greatest increase in mean CSA in both groups occurred between age 5 and 8 months. Across all age categories, there was no significant difference in mean CSA between the left and right limbs, or colts and fillies; there was a trend towards a larger CSA in the CONDEX group (P = 0.058). CONCLUSIONS: There was no conclusive evidence for a structural adaptive hypertrophy of the SDFT, probably because the regimen was insufficiently rigorous or because spontaneous pasture exercise may induce maximal development of energy storing tendons. POTENTIAL RELEVANCE: A moderate amount of early conditioning exercise against a background of constant exercise at pasture is not harmful to the development of the flexor tendons.

Evaluation of a new strategy to modulate skeletal development in racehorses by imposing track-based exercise during growth: the effects on 2- and 3-year-old racing careers.

REASON FOR PERFORMING STUDY: No data exist on the effects of conditioning exercise at foal age on workload and subsequent clinical injury rate during their 2- and 3-year-old racing careers. OBJECTIVES: To investigate the effects of subjecting TB foals to conditioning exercise prior to the start of race training on: the workload required to reach a level of fitness sufficient to compete; and the prevalence of orthopaedic injury during the first 2 seasons of their racing careers. METHODS: Twenty 18-month-old TBs, 12 subjected to conditioning exercise at foal age (CONDEX) and 8 exercised spontaneously at pasture only (PASTEX) were trained and entered in competitive events. Workload was quantified using the cumulative workload index (the product of average velocity and distance at a specific gait) and the animals were monitored clinically and radiographically for signs of musculoskeletal disorders. RESULTS: Workload to reach the desired fitness level was similar for CONDEX and PASTEX. CONDEX performed more prerace training sessions as 2-year-olds (P<0.05). The incidence of orthopaedic injuries was low in both groups and there were no differences in the occurrence of orthopaedic ailments. PASTEX animals tended to show signs of musculoskeletal disorders earlier than CONDEX animals. This time difference was significant for metacarpophalangeal joint pain on flexion, reduced carpal flexion and hindlimb lameness (P<0.05). CONCLUSIONS: Subjecting TB foals to conditioning exercise early in life does not have adverse effects on racing careers at ages 2 and 3 years, and does not influence the workload needed to reach a fitness level that is sufficient for racing. POTENTIAL RELEVANCE: The lack of negative effects and the indications of some positive effects of early conditioning exercise in the Thoroughbred encourage further large-scale comparative studies.

Evaluation of a new strategy to modulate skeletal development in Thoroughbred performance horses by imposing track-based exercise during growth.

REASON FOR PERFORMING STUDY: No data exist on the intensity of exercise required or on possible harmful effects of increasing exercise in foals over the natural level when free at pasture. OBJECTIVES: To investigate whether an increase in workload over free pasture exercise in the period from directly after birth to the start of training is tolerated by Thoroughbred (TB) foals without increasing injury rate or producing other undesired side effects. METHODS: Thirty-three TB foals were allocated to one of 2 exercise groups directly after birth. One group (PASTEX) was raised on pasture and the other (CONDEX) kept under identical circumstances, but was additionally subjected to an exercise protocol of gradually increasing intensity. Foals were monitored periodically and scored for the presence of clinical signs related to the musculoskeletal system (joint effusion, pain at flexion, occurrence of physeal swelling), and radiographs taken at the end of the conditioning phase. Also, behavioural studies were performed to detect any changes in behaviour related to the exercise programme. Cortisol levels were measured in both groups, to assess the level of stress. RESULTS: Workload in the CONDEX group was significantly higher than in the PASTEX group (approximately 30%). Conditioning increased the likelihood for joint effusion in the antebrachiocarpal joint, but reduced tarsocrural effusion and physeal swelling at the lateral distal radius, the third metacarpal bone (medial aspect) and lateral and medial aspects of the third metatarsal bone. CONCLUSIONS: The 30% increase in workload did not affect the animals' welfare, effects of conditioning exercise on clinical musculoskeletal health were few and there were no adverse effects. POTENTIAL RELEVANCE: This study supports the feasibility of imposing early conditioning exercise in horses and is a benchmark for its effects on the development of equine musculoskeletal tissues.

Early exercise advances the maturation of glycosaminoglycans and collagen in the extracellular matrix of articular cartilage in the horse.

REASON FOR PERFORMING STUDY: Training at a very young age may influence the characteristics of the collagen network of articular cartilage extracellular matrix (ECM) in horses. OBJECTIVES: To investigate whether increasing workload of foals results in significant changes in the biochemical composition of articular cartilage ECM. METHODS: Thoroughbred foals (n = 33) were divided into 2 different exercise groups from age 10 days-18 months. One group (PASTEX; n = 15) was reared at pasture; the other (CONDEX; n = 18) underwent a specific additional training programme that increased workload by 30%. At mean age 18 months, 6 animals from each group were subjected to euthanasia. The proximal articular surface of the proximal phalanx of the right hindlimb was examined for the presence of damage using the cartilage degeneration index (CDI). Samples were taken from 2 sites with known different loading patterns. Slices were analysed for DNA, glycosaminoglycans (GAG), collagen and post translational modifications of collagen (formation of hydroxylysylpyridinoline [HP] and pentosidine crosslinks, and hydroxylysine [Hyl]), and exercise groups and different sites compared. RESULTS: There were no differences in CDI between PASTEX and CONDEX animals, indicating the absence of extra joint damage due to the exercise regimen. There were site-related differences for most biochemical variables, corroborating earlier reports. All biochemical variables showed differences between PASTEX and CONDEX groups at one of the sites, and some at both. GAG and collagen levels were lower in the CONDEX group whereas Hyl, HP crosslinks and pentosidine crosslinks were higher. CONCLUSIONS AND POTENTIAL RELEVANCE: A measurable effect of the conditioning exercise was demonstrated. The margin between too much and too little work when training foals may be narrower than intuitively presumed.

Osteochondrosis and copper: histology of articular cartilage from foals out of copper supplemented and non-supplemented dams.

Copper (Cu) supplementation of dams in late gestation may be protective against articular cartilage abnormalities in foals. Articular cartilage was harvested from 22 Thoroughbred foals at 160 days of age, at sites predisposed to osteochondrosis (OC), and examined for evidence of early cartilage abnormalities and established dyschondroplastic (DCP) lesions to determine if there were any significant differences due to mare Cu supplementation by injection during late gestation, or foal liver Cu concentration. Cu supplemented mares received calcium Cu edetate injections in late gestation (250 mg at around 220, 248, 276 and 304 days gestation, then every two weeks until foaling). Foals were euthanased at 160 days of age and articular cartilage was harvested from four defined sites. Samples were examined for histological appearance of chondrocytes after staining with haematoxylin and eosin, and were also stained with toluidine blue to indicate proteoglycan content. Alkaline phosphatase (ALP) activity was detected by histochemistry, and histocytochemical techniques were used to determine the expression of cathepsin B. Cu supplementation of the dam, or liver Cu concentration of the foal at birth or 160 days of age had no statistically significant effect on the frequency of cartilage irregularities observed grossly, or abnormalities detected histologically at four defined sites. ALP expression was similar in all samples. Cathepsin B expression varied between sites, and was seen in chondrocyte clusters. The intensity of toludine blue staining varied between sites. Minor histological cartilage abnormalities were observed in cartilage from clinically normal animals. These abnormalities might be 'early' dyschondroplastic lesions, which could resolve or progress. The role of Cu in the development, resolution or progression of dyschondroplastic lesions is poorly understood.

Imposed running exercise does not alter cell proliferation in the neurogenic niches of young lambs.

Neurogenesis, the process by which neurons are generated in the brain from progenitor cells, occurs in the subventricular zone (SVZ) and the subgranular zone (SGZ) in the adult human brain. Recently, rodent studies have demonstrated that exercise can increase neurogenesis in the SGZ; however, it is unclear if exercise also has this effect in more complex mammalian brains. The overarching aim of this study was to explore whether exercised-induced neurogenesis occurs in larger mammalian brains more representative of human brains and to explore the use of a model for exercising large animals such as sheep. For these studies, 6 male twin lambs had a structured exercise regime for 4 wk and 6 other twin male lambs were kept in an open field pen. All lambs were injected with bromodeoxyuridine (BrdU), a thymidine analog that is incorporated into the DNA of proliferating cells. Immunoperoxidase was used to visualize and quantify BrdU-positive cells in the SVZ and SGZ. Overall, no significant change in the number or distribution of BrdU-positive cells was observed in the lamb SVZ and SGZ with exercise or colabeling of BrdU with mature neuronal or glial markers in the exercised and nonexercised lamb SVZ and SGZ. Overall, this study provides a novel methodology to investigate the effects of imposed exercise on large animals and exercise-induced neurogenesis in animals with gyrencephalic brains.

The pituitary and testicular responses to GnRH challenge between 4 and 14 months of age in thoroughbred colts born in spring and autumn.

Gonadotropin releasing-hormone analogue (buserelin) challenges were carried out every 8 weeks from 4 to 14 months of age on thoroughbred colts born in the spring (n = 6) or autumn (n = 5) to define the onset of puberty. In all colts, luteinizing hormone (LH) secretion followed a seasonal pattern, with high baseline and maximal concentrations in the spring and summer and low concentrations in the winter. Testosterone concentrations were undetectable before spring and, thus, autumn-born colts were younger than spring-born colts when a testosterone response to buserelin was first observed. Mean weights of the autumn-born colts were 300 kg (282-327 kg) at the time of the first detectable testosterone response in the following spring (October). Spring-born colts had reached this weight in the winter (May and June, before day length had increased) but did not exhibit a significant testosterone response until the spring at a mean weight of 352 kg (327-403 kg). It is proposed that colts must achieve a threshold body weight concurrently with stimulatory photoperiod for onset of puberty to occur.

Registration of confocal scanning laser microscopy and quantitative backscattered electron images for the temporospatial quantification of mineralization density in 18-month old thoroughbred racehorse articular calcified cartilage.

Combined backscattered electron scanning electron microscopy (BSE SEM) and confocal scanning laser microscopy (CSLM) have been used to put tissue mineralization data into the context of soft tissue histology and fluorescent label information. Mineralization density (Dm) and linear accretion rate (LAR) are quantifiable parameters associated with mineralizing fronts within calcified tissues. Quantitative BSE (qBSE) may be used to determine Dm, while CSLM may be used to detect label fluorescence from which LAR is calculated. Eighteen-month old Thoroughbred horses received single calcein injections 19 and 8 days prior to euthanasia, labeling sites of active mineralization with fluorescent bands. Confocal scanning laser microscopy images of articular calcified cartilage (ACC) from distal third metacarpal condyles were registered to qBSE images of the same sites using an in-house program. ImageJ and Sync Windows enabled the simultaneous collection of LAR and Dm data. The repeatability of the registration and measurement protocols was determined. Dm profiles between calcein labels were explored for an association with time. Dm was 119.7 +/- 24.5 (mean +/- standard deviation) gray levels (where 0 = backscattering from monobrominated and 255 from monoiodinated dimethacrylate standards, respectively), while modal and maximum LAR were 0.45 and 3.45 microm/day, respectively. Coefficients of variation (CV) for Dm were 0.70 and 0.77% with and without repeat registration, respectively; CVs for LAR were 1.90 and 2.26% with and without repeat registration, respectively. No relationship was identified between Dm and time in the 11-day interlabel interval. Registration of CSLM to qBSE images is sufficiently repeatable for quantitative studies of equine ACC.

Different short-term mild exercise modalities lead to differential effects on body composition in healthy prepubertal male rats.

Physical activity has a vital role in regulating and improving bone strength. Responsiveness of bone mass to exercise is age dependent with the prepubertal period suggested to be the most effective stage for interventions. There is a paucity of data on the effects of exercise on bone architecture and body composition when studied within the prepubertal period. We examined the effect of two forms of low-impact exercise on prepubertal changes in body composition and bone architecture. Weanling male rats were assigned to control (CON), bipedal stance (BPS), or wheel exercise (WEX) groups for 15 days until the onset of puberty. Distance travelled via WEX was recorded, food intake measured, and body composition quantified. Trabecular and cortical microarchitecture of the femur were determined by microcomputed tomography. WEX led to a higher lean mass and reduced fat mass compared to CON. WEX animals had greater femoral cortical cross-sectional thickness and closed porosity compared to CON. The different exercise modalities had no effect on body weight or food intake, but WEX significantly altered body composition and femoral microarchitecture. These data suggest that short-term mild voluntary exercise in normal prepubertal rats can alter body composition dependent upon the exercise modality.

Effects of racetrack exercise on third metacarpal and carpal bone of New Zealand thoroughbred horses.

The response of equine bone to training has not been quantified in racetrack trained horses, only in treadmill exercised horses. Seven two-year-old thoroughbred fillies were trained on sand and grass at a racetrack, in a typical New Zealand flatrace training regime. The horses were exercised 6 days per week for up to 13 weeks. During the day the horses were confined in 4 x 4m sand yards, and were stalled at night. Another 7 fillies of the same age were allowed free exercise in grass yards. The bones of the animals were available after the 13 week experimental period, and were examined using a Siemens Somatom AR CT scanner. To quantify the response of epiphyseal bone, 3mm thick sagittal plane images of the carpus (through the middle of the medial condyle of distal radius) and the distal third metacarpal bone (Mc3) (immediately lateral and medial to the junction of the condyle and the median sagittal ridge) were studied. Appropriate areas of interest were chosen, and the mean tissue density equivalent (Houndsfield Units) was determined. In the carpus, there was a significant effect of exercise in the dorso-distal aspect of the radius (p<0.01), dorsal aspect of radial and third carpal bones (p<0.01 and p<0.001 respectively). In palmaro-distal subchondral bone of Mc3, there was a significant effect on the medial/lateral site (p<0.01), which differed between right and left legs, probably due to the effect of the horses having been trained in one direction around the training track. The mean tissue density of the Mc3 epiphysis of the exercised group was 36.8% greater than that of the non-exercised group (p<0.001). The study demonstrates that bone response is both rapid and substantial, which should prompt the use of non-invasive diagnostic aids to determine the stage of training in which tissue density changes occur.