Genetic parameters of biokinematic variables of the trot in Spanish Purebred horses under experimental treadmill conditions.

The purpose of this study was to estimate the genetic parameters of biokinematic variables in Spanish Purebred (SPB) horses in order to select those of sufficient interest to be measured in the pre-selection of the animals for possible inclusion in the breeding programme. Kinematic analysis of 130 SPB horses 4.6+/-1.5 years old were recorded at the trot (4m/s) on a treadmill. Genetic parameters were estimated using VCE software and a bivariate mixed animal model including age and stud as fixed effects and animal additive genetic effect and residual error as random effects. In general, heritabilities were high (0.33-0.88). The angular variables presented the lowest heritabilities, whereas the maximum height of the fore-hoof and the duration of swing phase in the hindlimb gave the highest scores. Genetic correlations were also very high, so it was possible to reduce the number of breeding programme characteristics to stride duration, hindlimb swing phase duration, range of stifle and elbow angles, minimal angle of carpus, and minimal retraction-protraction angle of the hindlimb.

Degree of correspondence between contractile and oxidative capacities in horse muscle fibres: a histochemical study.

Samples taken from the middle gluteal muscle of 95 untrained adult horses of different ages and sex were subjected to histochemical analysis using the myosin adenosine triphosphatase (m-ATPase) and nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR) staining techniques. Fibres were classified into types I, IIA and IIB according to m-ATPase activity after preincubation at pH 4.4. The percentage of FT (Fast-Twitch Glycolytic) fibres and the proportion of IIB fibres with "high" and "low" oxidative capacity were determined in serial sections stained for NADH-TR. Statistical analysis revealed a significantly higher proportion of IIB fibres than FT fibres (P less than 0.001), though both percentages were correlated. Thus, 72.2 +/- 17.6% of type IIB fibres showed low oxidative capacity, but the remaining 27.8 +/- 17.6% showed high aerobic potential, and thus did not correspond to FT fibres. These results confirm that the contractile capacity of a muscle fibre does not determine its oxidative profile. The different types of muscle fibre should thus be classified solely according to m-ATPase activity, since this characteristic is related to the molecular structure of contractile proteins. Oxidative capacity should be assessed separately, and not be used as a criterion for fibre classification in horses.

Skeletal muscle histochemistry in male and female Andalusian and Arabian horses of different ages.

Muscle biopsies were taken from the middle gluteal muscle of 143 untrained horses (83 Andalusians [AN] and 60 Arabians [AR]) ranging from 10 days to 24 years old. The horses were separated according to breed and sex and allotted to five age groups: A, 0 to three months; B, yearlings; C, two to three years; D, five to 10 years; and E, 11 to 24 years. There was an increase in the percentage of type I fibres (about 100 per cent) as well as a decrease in the percentage of type IIB fibres (AN, 50 per cent; AR, 40 per cent) over the five age groups. The percentage of type IIA fibres rose significantly over the first two or three age groups. The overall decrease in the subgroup IIB fibres with age was proportionally greater for IIB oxidative fibres (AN, 72 per cent; AR, 68 per cent) than for IIB non-oxidative fibres (AN, 5 per cent; AR, 34 per cent). The mean cross-sectional area of all three fibre types increased significantly with age. In any given age group, the mean relative cross-sectional area occupied by IIA fibres in the biopsy specimens was significantly greater in stallions than in mares, at the expense of IIB fibres.

Kinematic characteristics of Andalusian, Arabian and Anglo-Arabian horses: a comparative study.

The aim of this study was to compare the kinematic trot characteristics of three different breeds of horse: Andalusian (AN, n = 15), Arabian (AR, n = 7) and Anglo-Arabian (AA, n = 5) using standard computer-assisted videography (25 Hz). Linear, temporal and angular parameters in fore- and hind limbs were analysed in six randomly selected strides per horse. Normalised angle-time diagrams along the complete stride were obtained for all joints angles in each breed and specific kinematic characteristics were detected graphically. AA horses displayed longer swing durations in both limbs ans a shorter angular range of motion (ARM) in scapula and pelvis inclination and in shoulder, hip and forelimb retraction-protraction angles. At lift off, stifle and tarsal joint angles were more flexed. In general, only small differences were observed in AR horse kinematics when compared with the other 2 breeds. AN horses presented negative overtracking length, which was positive in AR and AA. In AN horses the elbow and carpal joints were more flexed at the moment of maximal elevation, elbow and fore-fetlock joints also exhibited a larger ARM due to a smaller angle at maximal flexion. In the hind limbs, tarsal, hind fetlock and retraction-protraction angles presented a larger ARM in AN horses due to greater maximal flexion in the tarsal and hind fetlock joints. Fore- and hind fetlocks were also more flexed in horses from this breed. In conclusion, differences between kinematic variables at the trot were observed in the three breeds studied here, mainly in forelimb joints. The most outstanding feature was the greater forelimb flexion recorded in AN horses than in the other breeds which is consistent with the elevated movements in this breed. In AA horses, the ARM of proximal joints involved in retraction protraction in both fore- and hind limbs was smaller. All the differences observed highlighted the idiosyncratic nature of the trot in each breed; this may influence the functional capacity of each breed.

Muscle fibre type composition in untrained and endurance-trained Andalusian and Arab horses.

Muscle biopsies were taken from the middle gluteal muscle of 68 stallions (52 Andalusian [AN] and 16 Arab [AR]) ranging from six to 12 years of age. Seventeen AN horses and eight AR horses were untrained, while the remainder underwent active endurance training for 6 months. Fifteen AN horses were moderately endurance-trained while the other 20 AN horses and eight AR horses were strongly endurance-trained. Percentages of type I and type II fibres were similar in all groups (P greater than 0.05). The group of horses with the hardest training had a higher percentage of type IIA fibres (AN P less than 0.01; AR P less than 0.02) and a lower percentage of type IIB fibres (P less than 0.05 for AN and AR) than the untrained horses. All trained horses had a higher percentage of type IIB oxidative fibres and a lower percentage of type IIB non-oxidative fibres than the untrained horses (P less than 0.01 for both). These results suggest that the proportions of type I and type II fibres are highly stable within a given breed, but the stimulus of training facilitates changes both in contractile properties and, particularly, in the oxidative capacity of type II subgroups.

Angular kinematic patterns of limbs in elite and riding horses at trot.

Normal speed videography was used to determine the angular parameters of 28 Spanish Thoroughbreds at trot. Horses were divided into 3 groups: Group UT, comprising 9 animals (provided by the VII National Stud, Cordoba, Spain) which had undergone no specific training programme and which were hand led at the trot; Group T, formed by 19 horses considered to be highly bred and trained, and which were also hand led; and Group RT, comprising the same horses as the latter group but this time trotted by a rider. Each animal was filmed 6 times from the right-hand side, using a Hi8 (25 Hz) video camera. Angular parameters for fore- and hindlimb joints were measured in each stride from computer-grabbed frames and entered into a spreadsheet for calculation; parameters included maximum and minimum angles, range of motion, and angles at landing, lift off and maximum hoof height; the times at which maximum angle, minimum angle, lift off and maximum hoof height occurred were calculated as percentages of total stride duration. Stride velocity (mean [s.d.]) was 4.01 (0.62), 3.60 (0.34) and 3.07 (0.36) m/s for Groups UT, T and RT, respectively. Data were then compared between Groups UT-T and Groups T-RT. Compared with Group UT, horses from Group T featured a shorter stance percentage (P<0.001) in both fore- and hindlimbs. The range of motion in forelimbs was smaller (P<0.05), due to lower retraction (P<0.001); moreover, maximum retraction appeared earlier (P<0.05). Greater scapular inclination was in evidence (P<0.05) and the shoulder joint extended further (P<0.05). Fore- and hind fetlock joints revealed a relatively shorter hyperextension period during the stance phase (P<0.01). Compared with Group T, horses from Group RT had a longer stance percentage, with belated maximum retraction of the fore- and hindlimbs. The range of movement in scapular inclination was greater (P<0.05), due to a smaller minimum angle (P<0.01), and the shoulder joint flexed more (P<0.05). The elbow joint extended more and for longer during the stance phase. Initial extension of the hip joint (P<0.05) and tarsus (P<0.001) lasted longer. The carpal and fore and hind fetlock joints recorded relatively longer hyperextension times, in addition to greater hyperextension during the stance phase. The results from the present study suggest that rider-effect must be taken in consideration when well gaited horses are selected for dressage purposes.

The influence of speed and height at the withers on the kinematics of sound horses at the hand-led trot.

The influence of speed and height at the withers on some biokinematic stride parameters (linear, temporal and angular) was measured in 15 Spanish Thoroughbreds (Andalusian Purebred) trotted hand-led along a track; analysis was made of the correlation between speed and height at the withers and of some biokinematic parameters of equine locomotion. Both height at the withers and speed were positively and significantly correlated to the linear parameters examined. Temporal parameters did not reveal a significant correlation with the height at the withers. The correlations with the angular parameters were variable, often being significant for both height at withers and speed, although in most cases the coefficient was relatively low, probably owing to the narrow ranges of speed and height at the withers and their low variability. It was concluded that horses at the hand-led trot tend to have low variability in speed during successive trials, although the speed at which they move does have a significant influence on both linear and temporal stride parameters, and also on several angular parameters, while height at the withers has a major effect on linear parameters but less on angular parameters, and does not affect the temporal parameters.

Influence of training on the biokinematics in trotting Andalusian horses.

The aim of this study was to determine the influence of a 10-month training programme on the linear, temporal and angular characteristics of the fore and hind limbs at the trot in the Andalusian horse, using standard computer-aided videography. Sixteen male Andalusian horses were observed before and after training. Six strides were randomly selected for analysis in each horse and linear, temporal and angular parameters were calculated for fore and hind limbs. The training programme used here produced significant changes in kinematic parameters, such as shortening of stride length, and increase in swing duration and a decrease in hind limb stance percentage. No significant differences were recorded in the angular values for the forelimb joints. In trained horses, the more proximal joints of the hind limb, especially the hip and stifle, had a greater flexion while the fetlock showed a smaller extension angle. At the beginning of the swing phase, hip and stifle joints presented angles that were significantly more flexed. When the hind limbs came into contact with the ground, all the joints presented greater flexion after training.

Morphometric changes in the dog trochlear nerve with growth.

The objective of this work was to analyse changes in morphometric characteristics related to growth in the trochlear nerve in dogs. Twenty beagles, split into four dog age groups (A, 7 days; B, 21 days; C, 35 days; D, 49 days and E, 4 years), were used. The right intracranial portion of the nerve was analysed by light and electron microscopy. The nerve cross-sectional area was calculated. Number, diameter and cross-sectional area of unmyelinated and myelinated fibres were also calculated. In myelinated fibres, the corresponding axon area and diameter and myelin sheath thickness were also calculated. The number of myelinated and unmyelinated fibres was 1070.25±112.07 and 592.25±467.53 in group A, 1367±57.98 and 143.67±54.37 in group B, 1574.20±299.50 and 151.67±51.73 in group C, 1340.33±151 and 127±48.75 in group D and 1476±260.71 and 284±101.82 in group E. The mean diameter for myelinated and unmyelinated fibres was 4.37±0.17 µm and 0.41±0.08 µm for group A; 6.21±0.12 µm and 0.30±0.03 µm for B; 6.90±0.91 µm and 0.32±0.03 µm for C; 7.86±1.19 µm and 0.32±0.02 µm for D; 10.63±0.50 µm and 0.30±0.01 µm for E, respectively. This nerve possesses similar structural and ultrastructural features to the same nerve in other species and modifies its morphometry with growth. Results could enhance the understanding of pathological disorders.

Evaluation of conformation against traits associated with dressage ability in unridden Iberian horses at the trot.

The work describes and compares the conformation traits and gait characteristics in the main native Iberian dressage Purebreds (Lusitano, Spanish and Menorca horses). In total, 4 strides belonging to each of 66 males (4-14 years old) were recorded at handled trot, using a 3D semi-automatic movement analysis system. Principal component analysis (PCA) and intraclass Correlations (iC) were computed. The Menorca Purebred showed proportionally higher limbs (23.8 and 15.1cm for the forelimb cannon and pastern lengths), the Lusitano Purebred had larger inclinations (66.1°, 37.7° and 47.9° for the scapula and pastern angles), and the Spanish Purebred presented a longer scapula (41.5 cm) and a larger hip angle (132.1°), apparently showing a better conformation for dressage gaits. However, the three breeds presented undesirable tarsus joint conformation (<155.5°) for dressage performance. In general, all of them showed good features in the kinematics of the forelimbs. Nevertheless, the functional capacity of their hindlimbs and linear parameters probably need to be improved for their competitiveness.

2D versus 3D in the kinematic analysis of the horse at the trot.

The handled trot of three Lusitano Purebred stallions was analyzed by using 2D and 3D kinematical analysis methods. Using the same capture and analysis system, 2D and 3D data of some linear (stride length, maximal height of the hoof trajectories) and angular (angular range of motion, inclination of bone segments) variables were obtained. A paired Student T-test was performed in order to detect statistically significant differences between data resulting from the two methodologies With respect to the angular variables, there were significant differences in scapula inclination, shoulder angle, cannon inclination and protraction-retraction angle in the forelimb variables, but none of them were statistically different in the hind limb. Differences between the two methods were found in most of the linear variables analyzed.

Structural and ultrastructural study of the intracranial portion of the oculomotor, trochlear and abducent nerves in dog.

The right intracranial portion of the oculomotor, trochlear and abducent nerves were removed from six adult German shepherd dogs and analysed by light and electron microscopy. In all cases the nerve sectional area was calculated. Unmyelinated and myelinated fibres were analysed and number, diameter and cross-sectional area were calculated. In myelinated fibres, also calculated were the corresponding axon area and diameter, and myelin sheath thickness. The mean number of myelinated fibres was 8543.50 +/- 1231.85 being the unmyelinated 1402 +/- 241.58 in the oculomotor nerve; 1509 +/- 223.17 and 287.67 +/- 72.28 in the trochlear nerve and 2473.00 +/- 211.41 and 231.25 +/- 92.67 respectively in the abducent. The mean diameter was 10.23 +/- 0.68 microm in myelinated and 0.43 +/- 0.21 for unmyelinated in oculomotor nerve, 10.53 +/- 0.55 microm and 0.33 +/- 0.04 for the trochlear, and 10.45 +/- 1.27 microm and 0.47 +/- 0.09 in the abducent nerve respectively. This study reveals that oculomotor, trochlear and abducent nerves of the dog show structural and ultra-structural features similar to the same nerves in other species.

Variability of muscle fibre composition and fibre size in the horse gluteus medius: an enzyme-histochemical and morphometric study.

To determine the variability in fibre types and fibre sizes in the equine gluteus medius muscle, biopsy specimens were removed from 5 sites, at 4 different depths, within the right and left muscles of 3 Andalusian stallions. The percentage, lesser fibre diameter and cross-sectional area of the various fibre types were measured systematically in myosin ATPase and NADH-tetrazolium reductase-stained, serial cryostat sections of these multiple samples. Significant differences in muscle fibre type composition were recorded, with a lower percentage of type I fibres (high myosin ATPase activity at pH 4.5) being observed towards superficial regions of the muscle and a greater percentage towards the deep areas. Type II B fibres (moderate myosin ATPase activity at pH 4.5), including both II B nonoxidative (low NADH-TR activity) and II B oxidative (moderate NADH-TR activity), displayed the opposite tendency, and the percentage of type II A fibres (low myosin ATPase activity at pH 4.5) did not change with depth. Types I and II A fibres in the deep regions were larger than superficially, whereas the II B fibres in the deep regions were smaller than in the superficial parts of the muscle. The results also imply that type I fibres tend to be larger than type II fibres in the deep regions. The size of type I fibres is more homogeneous in the deep parts than in the superficial regions of the muscle, while II B fibres vary more in size in the peripheral portions than in deep regions. A single biopsy taken from the gluteus medius muscle of the horse is therefore a poor representative of the whole muscle and care should be exercised in sampling and interpreting data obtained from limited biopsy of this muscle. The pattern of variation in fibre types and fibre sizes between the different depths of the muscle probably reflect different functional demands on the gluteus medius muscle.

[Histochemical and morphometric characteristics of some extraocular muscles of the dog]. / Caracteristicas histoquimicas y morfometricas de algunos musculos extraoculares del perro.

This investigation was carried out on retractor bulbi, lateral and medial rectus muscles of six adult dogs. Tissues were collected from near the center of individual muscle bellies. These were stained for m-ATPase at varying pHs during preincubation, NADH-TR, Alpha-GPDH, Modified Mason trichrome and Hematoxyline and Eosin. Muscle fibers were classified as type I and type II, based on their reaction for m-ATPase. The retractor bulbi muscle was composed entirely of type II, oxidative, muscle fibers, with no glycolytic fibers. The rectus muscles presented a stratified composition, with superficial muscle fiber bundles containing a mixture of type II fibers oxidative and glycolytic, and central bundles of type I, mixed with type II. It was observed that there was less interstitial tissue in the center of the muscle bellies. Large diameter nerve fibers were also observed in the central layers.

Comparative biokinematic study of young and adult Andalusian horses at the trot.

The purpose of this study was to determine basic kinematic parameters (linear, temporal and angular) in young and adult Andalusian horses (P.R.E.) at the trot, using a normal computer-aided videography system. The trotting gaits of 16 horses were analysed: seven young horses (3.7 +/- 0.2 years old, height at withers 167.1 +/- 4.1 cm) and nine adult stallions (12.3 +/- 2.9 years old, height at withers 162.9 +/- 3.6 cm) were recorded at least 6 times at the trot using a 25-Hz video-camera filming from the side. Video images were processed with a real-time digital system (SMVD). Speeds averaged 3.84 and 3.75 m/s for young and adult horses, respectively. Differences between age groups for speed and linear and temporal parameters of the stride were not significant. However, variations in angular parameters were detected: adults showed a greater ARM than younger horses for most forelimb joints. In the hind limb, hip, stifle, and, to a lesser degree, the tarsal joint, a smaller degree of extension during the stance phase was observed in adult horses.

Variations in fibre size in the gastrocnemius muscle of rats undergoing training.

Sixty-six Wistar rats of both sexes were divided into six groups. One group was used as control, and the others underwent different training schedules. Muscle samples taken from the red and mixed portions of m. Gastrocnemius (caput lateralis) were stained with m-ATPase in order to identify fibre types I, IIA and IIB. These fibres were measured in order to determine their minimum diameter. The most notable results obtained were: a) the three types of fibre were generally larger in males than in females; b) the mean minimum diameter of fibres from the mixed portion was smaller than those of the red portion, except type IIB fibres, and c) the training schedules involved generally led to slight hypertrophy of all types of fibres, although this finding was more significant in the mixed portion of m. Gastrocnemius.

Effects of training on fibre composition in rat gastrocnemius muscle.

Forty-eight adult Wistar rats were divided into four groups for experimentation. The group I was used as a control and groups II, III and IV underwent progressive treadmill training. Samples of the red and mixed portions of m. gastrocnemius (lateral head) were stained with the histochemical technique of m-ATPase to determine the percentage of type I, IIA and IIB fibres, and with NADH-TR, in order to quantify variations in "low-oxidative" fibre percentages. The results showed that progressive training did not lead to statistical variations in the percentage of type I fibres. However, the proportion of type IIA fibres rose, while that of IIB fibres fell, in both cases significantly. Variations were more marked in mixed than in red Gastrocnemius muscle. A clear decrease was noted in "low-oxidative" fibres, which were virtually absent from red portion. This decrease was more marked, and occurred more rapidly, than in type IIB fibres.

Kinematics in horses at the trot before and after an induced forelimb supporting lameness.

The aim of this study was to analyse the biokinematic alterations caused by an induced lameness in the right forelimb of Dutch Warmblood (DWB) horses using a system of computer-aided normal videography. Five mature DWB were recorded with a videocamera (frame rate 1/25) from a lateral view before and after an induced lameness. Before videotaping, passive markers were placed on the skin, over easily identifiable anatomical references to determine the joint angles in the forelimb (always on the flexor side). Lameness was induced using special horseshoes. The lameness was evident at the trot and mild at walk. The images were analysed using a real time digitalising system combined with a previously designed spreadsheet. Linear, temporal and angular parameters (maximum, minimum and angular range of motion) along the stride were calculated as well as the moments of highest extension (Pmax) and flexion (Pmin), expressed as a percentage of the whole stride. Results before and after the induced lameness were compared by a paired Student's t test at a significance level of P<0.05. No differences in speeds before and after the induced lameness were found. Stride length was significantly shorter in the lameness condition. Stride duration was slightly shorter in lameness. The diagonal stance phase increased, while the swing phase decreased. Angular parameters changed mainly in elbow, carpus, fetlock and retraction-protraction angles. This indicated that the angular range of motion in the elbow and carpal joints decreased, and the elbow Pmin occurred later in the stride. The results are useful in the development of video-based equine lameness diagnostics.

Inter-breed differences in equine forelimb kinematics at the walk.

Linear, temporal and angular biokinematic characteristics of the forelimb at the walk in different breeds were determined, highlighting inter-breed differences. Twenty-three healthy stallions were used: ten Andalusians (AN), seven Arabs (AR) and six Anglo-Arabs (AA). Height at the withers was significantly different between groups (P < 0.001). Six trials per horse were recorded using a levelled video camera (sampling frame rate 25 frames/s), digitized and analysed using a semi-automatic movement analysis system. No statistically significant differences in speeds were recorded between breeds (P > 0.05). The only temporal parameter which was similar in the three breeds was the moment at which the hoof reached the highest point in its trajectory. The variables presenting the most significant differences were the percentages of deceleration and propulsion within the stance phase. ANOVA for angular variables showed that the greatest difference was in the range of angular movement of the carpal joint, being higher in AN, due to a lower minimum value. In the fetlock joint, the greatest difference was observed in minimum values, which differed in all three breeds. Significant inter-breed differences were also observed for maximum limb retraction, being lowest in the AN group, followed by the AA and AR groups. This finding was reflected in the angular range of motion, despite smaller differences in the degree of limb protraction; very similar values were reported in all three breeds. As regards the elbow joint, no inter-breed differences were observed in terms of minimum values, whereas differences were recorded for maximum and angular range of motion, higher values being displayed by the AR and AN groups than by AA animals. In conclusion, inter-breed differences may be determined in equine forelimb biokinematics at the walk. This study distinguished between AN, AR and AA horses.

Changes due to age in the kinematics of trotting Andalusian foals.

The purpose of this study was to evaluate the effect of growth on biokinematic characteristics at the trot in a group of 9 Andalusian foals from age 12 to 36 months in order to identify which stride variables change or remain invariable. Biokinematic analysis was performed by using a computer-assisted videography system (25 Hz frame rate). An increase in stride length as well as the fore- and hindlimb stride duration was found. A tendency to increase flexion of the shoulder, elbow and carpal joints was observed while the forelimb fetlock increased its maximal extension. In the hindlimb, most of the differences were exhibited by the hip and stifle joints, which showed a tendency to a lesser flexion with age. However, at landing the extension was bigger. The hindlimb fetlock extended more in older foals, whereas flexion decreased. Significant differences in the maximal extension and flexion instants in the forelimb joints were not detected (except in the elbow joint). The same was found in the angular range of motion (ARM) of the scapula inclination and the shoulder and forelimb fetlock joints. Differences in retraction-protraction angles in both the fore- and hindlimbs were very slight, and ARM values for these parameters were similar in all age groups. It can be concluded that Andalusian horses modified their locomotor pattern at the trot between age 12 and 36 months, particularly up to 24 months, by increasing stride length as well as shoulder, elbow and carpus flexion, while hip and stifle flexion decreased and hindlimb joints were more extended during the stance phase. One of the most striking features was that maximal retraction and protraction angles did not change in foals age 12-36 months in either fore- or hindlimbs. These results could be useful for the prediction of some kinematic parameters of mature horses from data obtained at younger ages.