Post-thaw quality of ram sperm frozen with different concentrations of low-density lipoproteins associated with non-enzymatic antioxidants

The cryopreservation reduces ram sperm quality, decreasing the pregnancy rate of ewes inseminated with thawed sperm. Hence, we aimed to improve the post-thaw quality of ram sperm replacing egg yolk on Tris-Glucose extender with different concentrations of LDL (2 or 8%), associated with the addition of 10 mM non-enzymatic antioxidants (ascorbic acid, hydroxytoluene butylate, ascorbyl palmitate, and trehalose). Semen samples were collected from six rams, split into different treatments, and frozen. After thawing, kinematic (CASA), structural (propidium iodide and carboxyfluorescein diacetate) and functional (hypoosmotic test) sperm membrane integrity was assessed. Total motility, VCL, and LIN were also assessed in thawed samples during 3 h of incubation (38 °C). The results showed that hydroxytoluene butylate at 10 mM in Tris-Glucose extender with 8% LDL improved velocity parameters immediately post-thaw compared with Tris-Glucose egg yolk extender, as well as prevented the reduction of total motility and VCL after incubation. There was no benefit of adding ascorbic acid and trehalose. Moreover, for the first time, it was shown the motility impairment promoted by ascorbyl palmitate to ram sperm.(AU)

Influence of thawing temperature on sperm motility, structure, and metabolism of frozen bovine semen / Influência da temperatura de descongelamento na motilidade, estrutura e metabolismo do sêmen bovino congelado

The heating rate used during semen thawing plays an important role in reducing structural and functional damage to spermatozoa. In this study, we evaluated the influence of thawing temperature on semen quality, reactive oxygen species (ROS) production, and mitochondrial activity of cryopreserved bovine semen. A total of 195 straws of 0.5 mL from five Holstein Friesian bulls were used (39 straws per bull). Samples underwent 8 to 22 years of storage; they were processed under a standard protocol with tris-egg yolk and stored in liquid nitrogen. Samples were thawed for 30 seconds in a water bath at T1: 36 °C, T2: 38 °C or T3: 40 °C. Sperm motility and kinematics, morphology, structural membrane integrity (SMI), functional membrane integrity (FMI), acrosome integrity (AI), ROS, and mitochondrial membrane potential (ΔΨM) of post-thawing bovine sperm were evaluated. Generalized linear models were fitted to the data. Each model included the effects of bull, storage time, and treatment. The Shapiro-Wilk test was used to assess data normality, and means were compared using the Tukey test. T2 and T3 showed better results for sperm motility and kinematic parameters, SMI (%) (T1 41.9 ± 2.3; T2 45.7 ± 1.9; T3 47.4 ± 2.8), ROS (RFU/min) (T1 0.026 ± 0.007; T2 0.032 ± 0.001; T3 0.031 ± 0.001) and high-ΔΨM (RFU x 103) (67.1± 0,4; 71.3 ± 0.4; 74.2 ± 0.4) (P < 0.05). However, T1 had higher FMI (39.3 ± 2.3) than T2 (34.0 ± 1.9) (P < 0.05), though not significantly (P > 0.05) different from T3 (38.4 ± 2.2). Thawing temperatures of 38 °C and 40 °C increases motility, kinetics, membrane integrity, mitochondrial activity and ROS of cryopreserved bovine semen, compared with more conventional thawing at 36 °C.

A taxa de aquecimento usada durante o descongelamento do sêmen desempenha um papel importante na redução dos danos estruturais e funcionais nos espermatozóides. O objetivo desta pesquisa foi avaliar a influência da temperatura de descongelamento na qualidade do sêmen, produção de espécies reativas de oxigênio (ROS) e atividade mitocondrial do sêmen bovino criopreservado. Foram utilizados 195 palhetas de 0,5 mL de cinco touros Holstein Friesian (39 palhetas por touro). As amostras passaram por oito a 22 anos de armazenamento e foram processadas sob protocolo padrão com Tris-gema de ovo e armazenadas em nitrogênio líquido. As temperaturas de descongelamento foram T1: 36 °C, T2: 38 °C, T3: 40 °C, cada uma por 30 segundos em banho-maria. Pós-descongelamento, a motilidade e cinética dos espermatozoides, morfologia, integridade estrutural da membrana (SMI), integridade funcional da membrana (FMI), integridade acrossomal (AI), ROS e potencial de membrana mitocondrial (ΔΨM) foram avaliados. Modelos lineares generalizados foram ajustados. Cada modelo incluiu os efeitos de touro, tempo de armazenamento e tratamento. A normalidade dos dados foi avaliada pelo teste de Shapiro-Wilk e as médias comparadas pelo teste de Tukey. T2 e T3 apresentaram resultados mais elevados para a maioria dos parâmetros de motilidade e cinemática espermática, SMI (%) (T1 41,9 ± 2,3; T2 45,7 ± 1,9; T3 47,4 ± 2,8), ROS (RFU/min) (T1 0,026 ± 0,007; T2 0,032 ± 0,001; T3 0,031 ± 0,001) e alto ΔΨM (RFU x 103) (67,1 ± 0,4; 71,3 ± 0,4; 74,2 ± 0,4) (P < 0,05). No entanto, T1 apresentou maior FMI (%) (39,3 ± 2,3) em comparação a T2 (34,0 ± 1,9) (P < 0,05), mas não foi diferente do T3 (38,4 ± 2,2) (P > 0,05). Conclui-se que as temperaturas de descongelamento de 38 °C e 40 °C produzem um aumento na motilidade, cinética, integridade de membrana, atividade mitocondrial e ROS do sêmen bovino criopreservado, em comparação com o uso mais convencional de uma temperatura de descongelamento de 36 °C.

DNA methylation level of gene SIRT1 in ram spermatozoa and relationship with fertilizing ability according to breed and age

Background: Effect of the epigenetic factors on the male fertility is well proofed. Sperm acts as a carrier of genetic material, and its DNA methylome can affect maternal pregnancy rate and offspring phenotype. However, the research on the DNA methylation in the spermatozoids of livestock males, in particular rams, is still limited. To best of our knowledge the data about as a global as well as gene specific DNA methylation in ram spermatozoa from different breeds and ages are missed in the scientific literature. The present study was designed to analyze the relationship between methylation levels of the important for spermatogenesis gene SIRT1 in spermatozoa and fertilizing ability of sperm in rams from different breeds and ages. Materials, Methods & Results: The ejaculates of 16 rams from Lacaune, East Friesian and Assaf breeds at age between 18 to 96 months were evaluated. The kinematic parameters of 2 semen samples from each animal were estimated by CASA. The separated spermatozoa were used for DNA extraction followed by bisulfite conversion. The DNA methylation of SIRT1 was detected through quantitative methylation-specific PCR using 2 sets of primers designed specifically for bisulfite-converted DNA sequences to attach methylated and unmethylated sites. The breed and age effect on the gene SIRT1 methylation by ANOVA was estimated. Experimental females included 393 clinically healthy milk ewes (Lacaune, n = 131; East Friesian sheep, n = 100 and Assaf, n = 162) in breeding season. Reproductive performances (conception rate at lambing, lambing percentage and fecundity) of ewes, inseminated by sperm of the investigated rams, were statistically processed. ANOVA showed that the animal breed influences significantly on the level of DNA methylation of gene SIRT1 in ram spermatozoa (P = 0.002) An average value of DNA methylation of SIRT1 in ram sperm from Lacaune breed was significantly higher than in Assaf and East Friesian (81.21 ± 15.1% vs 36.7 ± 14.2% and 38.3 ± 18.6 respectively, P < 0.01). The highest percent of SIRT1 methylation was observed in old animals compared to the young and middle-age. Moderate and strong correlations (r from 0.44 to 0.71, P < 0.05) between the methylation level of the SIRT1 gene in rams' sperm and reproductive parameters of inseminated ewes in all breeds were established. Discussion: Our data are the first message about the effect of breed on the specificity of DNA methylation of gene SIRT1 in ram spermatozoa. These results demonstrated an existence of the sheep breeds with high and low level of DNA methylation of gene SIRT1 in ram sperm. Although the effect of age on the methylation level in sperm is still discussable, our results showed a moderate correlation between age and methylation level of SIRT1 in spermatozoa of rams. Taking into account that DNA methylation in sperm is stabilized with puberty onset and is a heritable epigenetic modification, it can be a promising marker of sperm quality in animal breeding. In all investigated breeds the rams with relatively high level of DNA methylation of gene SIRT1 in spermatozoa (50-68%) demonstrated a high conception rate at lambing (> 70%). In conclusion, the DNA methylation level of the SIRT1 gene in ram spermatozoa is determined by both the breed and the age of the animals and correlates with fertilizing ability of sperm.

Action of swim-up and caffeine on equine frozen sperm

Cryopreservation of equine semen is crucial to semen commercialization. However, it reduces sperm motility and longevity. Thus, sperm selection methods and addition of motility-activating substances to sperm, such as caffeine, may improve sperm quality of equine frozen semen. The objective of the current work was to evaluate the effects of caffeine on recovery and quality parameters of frozen-thawed sperm subjected to swim-up selection to be used in intracytoplasmic sperm injection (ICSI) in assisted reproductive techniques. Stallion semen were frozen and after thawing different caffeine concentrations were added to the samples performing four treatments control (no caffeine), 3, 5, and 7.5 mM caffeine. Sperm kinematic and motility were assessed by computer-assisted sperm analysis (CASA). Then, the four treated samples were submitted to the swim-up sperm selection, and the number of recovered sperm and morphology were evaluated at four times 20, 40, 60, and 80 min. The swim-up increased the recovery proportion of normal morphology sperm without (80.1±1%) or with caffeine addition (3mM: 81.2±1%, 5mM: 79.9±1% and 7.5 mM 78.9±1%) compared to the thawed semen (70±2%). However, the addition of 5 mM caffeine induced an increase in sperm motility (38.9±2.8 vs. 32.6±3.4%, P<0.05), and sperm recovery after swim-up (7.9x106 vs. 3.4x106 sperm/ml, P<0.05) compared to the control. The addition of 5 mM caffeine to frozen-thawed equine semen before swim-up selection improved sperm motility and increased the sperm recovery rate while not decreasing the percentage of morphologically normal sperm. Thus, caffeine addition to frozen-thawed equine semen before swim-up selection has potential clinical application in improving sperm quality for use in ICSI.(AU)

Two-dimensional and three-dimensional techniques for determining the kinematic patterns for hindlimb obstacle avoidance during sheep locomotion / Técnicas bidimensionais e tridimensionais para determinar os padrões cinemáticos durante a ultrapassagem de obstáculos pelos membros pélvicosdurante a locomoção de ovinos

Analysis of locomotion is often used as a measure for impairment and recovery following experimental peripheral nerve injury. Compared to rodents, sheep offer several advantages for studying peripheral nerve regeneration. In the present study, we compared for the first time, two-dimensional (2D) and three-dimensional (3D) hindlimb kinematics during obstacle avoidance in the ovine model. This study obtained kinematic data to serve as a template for an objective assessment of the ankle joint motion in future studies of common peroneal nerve (CP) injury and repair in the ovine model. The strategy used by the sheep to bring the hindlimb over a moderately high obstacle, set to 10% of its hindlimb length, was pronounced knee, ankle and metatarsophalangeal flexion when approaching and clearing the obstacle. Despite the overall time course kinematic patterns about the hip, knee, ankle, and metatarsophalangeal were identical, we found significant differences between values of the 2D and 3D joint angular motion. Our results showed that the most apparent changes that occurred during the gait cycle were for the ankle (2D-measured STANCEmax: 157±2.4 degrees vs. 3D-measured STANCEmax: 151±1.2 degrees; P .05) and metatarsophalangeal joints (2D-measured STANCEmin: 151±2.2 degrees vs. 3D-measured STANCEmin: 162 ± 2.2 degrees; P .01 and 2D-measured TO: 163±4.9 degrees vs. 3D-measured TO: 177±1.4 degrees; P .05), whereas the hip and knee joints were much less affected. Data and techniques described here are useful for an objective assessment of altered gait after CP injury and repairin an ovine model.(AU)

A análise da locomoção é frequentemente usada como uma medida para avaliar a disfunção e sua recuperação após lesão nervosa periférica experimental. Quando comparadas com os roedores, as ovelhas oferecem várias características atrativas como modelo experimental para o estudo da regeneração nervosa periférica. Não existem estudos acerca dos resultados da locomoção após lesão e reparação do nervo periférico no modelo ovino. No presente estudo, realizámos e comparámos a cinemática bidimensional (2D) e, pela primeira vez, tridimensional (3D) do membro pélvico durante a ultrapassagem de obstáculos no modelo ovino. Este estudo teve como objetivo obter dados cinemáticos para servir de modelo para uma avaliação objetiva do movimento articular do tornozelo em estudos futuros de lesão e reparação do nervo fibular comum (FC) no modelo ovino. A estratégia usada pelas ovelhas para elevar o membro pélvico sobre um obstáculo com uma altura moderada, fixado em 10% do seu comprimento, caracteriza-se por uma flexão pronunciada do joelho, tornozelo e metatarso-falangeana ao se aproximar e ultrapassar o obstáculo. Apesar dos padrões cinemáticos do quadril, joelho, tornozelo e metatarso-falangeano terem sido idênticos, foram encontradas diferenças significativas entre os valores do movimento angular das articulações em 2D e 3D. Os nossos resultados mostram que as mudanças mais aparentes que ocorreram durante o ciclo da marcha foram nas articulações do tornozelo (em 2DSTANCEmax: 157±2.4 graus vs. em 3D STANCEmax: 151±1.2 graus; P .05) e metatarso-falangeana (em 2D STANCEmin: 151±2.2 graus vs. em 3D STANCEmin: 162 ± 2.2 graus; P .01 e em 2D TO: 163±4.9 graus vs. em 3D TO: 177±1.4 graus; P .05), enquanto as articulações do quadril e do joelho foram muito menos afetadas. É provável que os dados e técnicas descritas aqui sejam úteis para uma avaliação objetiva das alterações na marcha após lesão e reparação do PC no modelo ovino.(AU)

Low Density Lipoprotein - important player in increasing cryoprotective efficiency of soybean lecithin-based bull semen extenders

Currently, considering cryopreservation of bull semen, there is no clear consensus over the comparability of cryoprotective efficacy of extenders with soybean lecithin and those based on egg yolk. The objective of this study was to prove the use of Low Density Lipoprotein (LDL) extracted from hen-egg yolk as an enhancing factor for soybean lecithin-based extenders. In total, 35 ejaculates of (seven bulls x five ejaculates per bull) were collected and cryopreserved at a commercial insemination centre. The effect of the LDL addition to the extenders AndroMed® and Bioxcell® was tested in a 6% (v/v) concentration on spermatozoa after thawing. Modified extender composition effects were assessed on sperm functional parameters motility, plasma membrane, mitochondrial membrane potential and acrosomal integrity after thawing by CASA, flow cytometry and fluorescent microscopy, respectively. Based on kinematic parameters determined from CASA, k-means cluster analysis was used to classify individual spermatozoon into specific subpopulations (fast, medium fast and slow). A subpopulation of fast spermatozoa was increased in the presence of LDL in both selected extenders (P 0.05). The percentage of sperm with intact acrosome was improved when LDL was added to Bioxcell® extender (P < 0.05). On the other hand, addition of LDL to AndroMed® extender improved mitochondrial intactness after thawing (P < 0.05). In conclusion, our results showed that adding LDL to selected soybean lecithin-based extenders considerably ameliorated the functional parameters of spermatozoa after thawing and thus this lipoprotein could represent an improving agent for soybean lecithin-based extender for bull semen.

Low Density Lipoprotein - important player in increasing cryoprotective efficiency of soybean lecithin-based bull semen extenders

Currently, considering cryopreservation of bull semen, there is no clear consensus over the comparability of cryoprotective efficacy of extenders with soybean lecithin and those based on egg yolk. The objective of this study was to prove the use of Low Density Lipoprotein (LDL) extracted from hen-egg yolk as an enhancing factor for soybean lecithin-based extenders. In total, 35 ejaculates of (seven bulls x five ejaculates per bull) were collected and cryopreserved at a commercial insemination centre. The effect of the LDL addition to the extenders AndroMed® and Bioxcell® was tested in a 6% (v/v) concentration on spermatozoa after thawing. Modified extender composition effects were assessed on sperm functional parameters motility, plasma membrane, mitochondrial membrane potential and acrosomal integrity after thawing by CASA, flow cytometry and fluorescent microscopy, respectively. Based on kinematic parameters determined from CASA, k-means cluster analysis was used to classify individual spermatozoon into specific subpopulations (fast, medium fast and slow). A subpopulation of fast spermatozoa was increased in the presence of LDL in both selected extenders (P < 0.05). Moreover, the positive effect of LDL on sperm motility was confirmed by decreasing the percentage of sperm in slow subpopulation (P < 0.05). The effect of LDL addition on the incidence of spermatozoa with intact plasma membrane was not demonstrated in any case of extender used (P > 0.05). The percentage of sperm with intact acrosome was improved when LDL was added to Bioxcell® extender (P < 0.05). On the other hand, addition of LDL to AndroMed® extender improved mitochondrial intactness after thawing (P < 0.05). In conclusion, our results showed that adding LDL to selected soybean lecithin-based extenders considerably ameliorated the functional parameters of spermatozoa after thawing and thus this lipoprotein could represent an improving agent for soybean lecithin-based extender for bull semen.(AU)

Biomechanical analysis of the masticatory movement before and after adjusting dental occlusion in equine / Análise biomecânica do movimento mastigatório antes e depois do ajuste oclusal dental em equinos

The aim of this study was to evaluate through three-dimensional kinematic analysis the influence of occlusal adjustment by tooth wear in masticatory biomechanics of horses. Seven clinically healthy thoroughbred Arabian horses with strong occlusal irregularities were used, of which seven castrated males and one female of between seven and nineteen years of age. Three digital video cameras and seven spherical reflective markers placed on the horses' face were employed. The animals were filmed twice in succession: while chewing hay before and after the occlusal adjustment by tooth wear. Following that, kinematic analysis was made of the movements obtained by means of the images which were synchronized, segmented and reconstructed three-dimensionally with the help of Dvideow program. Mathematical functions were applied in Matlab environment for obtaining the values of the amplitudes of movements. The results of the biomechanical analysis showed that the occlusal adjustment increases the range of mandibular movements in horses.(AU)

O objetivo deste estudo foi avaliar, através de análise cinemática tridimensional, a influência do ajuste oclusal por desgaste dentário na biomecânica da mastigação de equinos. Foram utilizados sete cavalos puro sangue árabe, clinicamente saudáveis, com irregularidades oclusais relevantes, sendo seis machos castrados e uma fêmea, de sete a dezenove anos de idade. Três câmeras de vídeo digitais e sete marcadores reflexivos esféricos foram colocados na face dos equinos. Os animais foram filmados duas vezes seguidas: durante a mastigação do feno antes e após o ajuste oclusal por desgaste dental. Na sequência, procedeu-se a cinemática dos movimentos obtidos por meio das imagens sincronizadas, segmentadas e reconstruídas em três dimensões, com a ajuda do programa Dvideow. Funções matemáticas foram aplicadas no ambiente Matlab para a obtenção dos valores de amplitudes dos movimentos. Os resultados da análise biomecânica mostraram que o ajuste oclusal aumenta a gama de movimentos mandibulares em equinos.(AU)

Biomechanical analysis of the masticatory movement before and after adjusting dental occlusion in equine / Análise biomecânica do movimento mastigatório antes e depois do ajuste oclusal dental em equinos

The aim of this study was to evaluate through three-dimensional kinematic analysis the influence of occlusal adjustment by tooth wear in masticatory biomechanics of horses. Seven clinically healthy thoroughbred Arabian horses with strong occlusal irregularities were used, of which seven castrated males and one female of between seven and nineteen years of age. Three digital video cameras and seven spherical reflective markers placed on the horses' face were employed. The animals were filmed twice in succession: while chewing hay before and after the occlusal adjustment by tooth wear. Following that, kinematic analysis was made of the movements obtained by means of the images which were synchronized, segmented and reconstructed three-dimensionally with the help of Dvideow program. Mathematical functions were applied in Matlab environment for obtaining the values of the amplitudes of movements. The results of the biomechanical analysis showed that the occlusal adjustment increases the range of mandibular movements in horses.(AU)

O objetivo deste estudo foi avaliar, através de análise cinemática tridimensional, a influência do ajuste oclusal por desgaste dentário na biomecânica da mastigação de equinos. Foram utilizados sete cavalos puro sangue árabe, clinicamente saudáveis, com irregularidades oclusais relevantes, sendo seis machos castrados e uma fêmea, de sete a dezenove anos de idade. Três câmeras de vídeo digitais e sete marcadores reflexivos esféricos foram colocados na face dos equinos. Os animais foram filmados duas vezes seguidas: durante a mastigação do feno antes e após o ajuste oclusal por desgaste dental. Na sequência, procedeu-se a cinemática dos movimentos obtidos por meio das imagens sincronizadas, segmentadas e reconstruídas em três dimensões, com a ajuda do programa Dvideow. Funções matemáticas foram aplicadas no ambiente Matlab para a obtenção dos valores de amplitudes dos movimentos. Os resultados da análise biomecânica mostraram que o ajuste oclusal aumenta a gama de movimentos mandibulares em equinos.(AU)

Analysis and characterization of the normal gait phases of walking Warmblood horses as a tool for the diagnosis of lameness / Análise e caracterização das fases do andamento normal dos cavalos através da combinação de fotogrametria e acelerometria como ferramenta para o diagnóstico da claudicação

Horses with lameness modify gait behavior, but when it is subtle, it may not be possible to identify it clinically. The objective of this research is to characterize the normal gait phases of walking Warmblood horses by combining photogrammetry and accelerometry to monitor lameness to indicate a structural or functional disorder in the extremities. The study was conducted in 23 adult male Warmblood horses. Photogrammetry was used to identify the kinematic variables of the limbs and the markers path over time; triaxial accelerometers were used to capture the orthogonal acceleration components. It was determined that only 10 horses showed a normal gait pattern, there was a 43% correspondence between the expert´s judgment and the diagnostic techniques. According to the Stashak classification of the gait phases, cycle phases to forelimb were 34/4/8/13/41, while for hind limb were 54/11/8/8/19 (% of the stride). The range of motion (ROM) of the neck, knee and fetlock joints was 45.52±5.63°, 196.04±19.7° and 209±11.52° respectively. A combination of experimental methods was used to identify the phases of gait cycle of healthy horses. There was a correspondence in the location of the points of maximum displacement of the limbs with both techniques. More detailed information on the limbs movement was obtained using the accelerometer technology. These methods are applicable to other conditions either outdoors or in the lab.(AU)

Os cavalos com claudicação modificam o comportamento do andamento. Porém, uma leve modificação pode não ser identificada clinicamente. O objetivo desta pesquisa foi caracterizar as fases do andamento normal dos cavalos através da combinação de fotogrametria e acelerometria, para identificar alterações estruturais ou funcionais nos membros. O estudo foi conduzido em 23 cavalos Warmblood machos adultos. A partir da fotogrametria foi possível obter as variáveis cinemáticas das extremidades e a trajetória dos marcadores ao longo do tempo. Por outro lado, os acelerômetros triaxiais foram usados para capturar as componentes ortogonais da aceleração. Determinou-se que apenas 10 cavalos mostraram um padrão de marcha normal, e houve uma correspondência de 43% entre pareceres dos peritos e as técnicas de diagnóstico. De acordo com a classificação das fases da marcha de Stashak, as fases do ciclo da extremidade anterior foram 34/4/8/13/41 enquanto para a extremidade posterior foram 54/11/8/8/19. A amplitude de movimento (ROM) de pescoço, joelho e sesamóide foram 45,52±5,63°, 196,04±19,7° e 209±11,52°, respectivamente. Uma combinação dos métodos experimentais foi utilizada para identificar as fases do ciclo de marcha de cavalos saudáveis. Constatou-se correspondência na localização dos pontos de deslocamento máximo do membro com ambas as técnicas. A informação mais detalhada do movimento dos membros foi obtida usando a acelerometria. Estes métodos são aplicáveis em outras condições, quer a campo ou no laboratório.(AU)

Analysis and characterization of the normal gait phases of walking Warmblood horses as a tool for the diagnosis of lameness / Análise e caracterização das fases do andamento normal dos cavalos através da combinação de fotogrametria e acelerometria como ferramenta para o diagnóstico da claudicação

Horses with lameness modify gait behavior, but when it is subtle, it may not be possible to identify it clinically. The objective of this research is to characterize the normal gait phases of walking Warmblood horses by combining photogrammetry and accelerometry to monitor lameness to indicate a structural or functional disorder in the extremities. The study was conducted in 23 adult male Warmblood horses. Photogrammetry was used to identify the kinematic variables of the limbs and the markers path over time; triaxial accelerometers were used to capture the orthogonal acceleration components. It was determined that only 10 horses showed a normal gait pattern, there was a 43% correspondence between the expert´s judgment and the diagnostic techniques. According to the Stashak classification of the gait phases, cycle phases to forelimb were 34/4/8/13/41, while for hind limb were 54/11/8/8/19 (% of the stride). The range of motion (ROM) of the neck, knee and fetlock joints was 45.52±5.63°, 196.04±19.7° and 209±11.52° respectively. A combination of experimental methods was used to identify the phases of gait cycle of healthy horses. There was a correspondence in the location of the points of maximum displacement of the limbs with both techniques. More detailed information on the limbs movement was obtained using the accelerometer technology. These methods are applicable to other conditions either outdoors or in the lab.(AU)

Os cavalos com claudicação modificam o comportamento do andamento. Porém, uma leve modificação pode não ser identificada clinicamente. O objetivo desta pesquisa foi caracterizar as fases do andamento normal dos cavalos através da combinação de fotogrametria e acelerometria, para identificar alterações estruturais ou funcionais nos membros. O estudo foi conduzido em 23 cavalos Warmblood machos adultos. A partir da fotogrametria foi possível obter as variáveis cinemáticas das extremidades e a trajetória dos marcadores ao longo do tempo. Por outro lado, os acelerômetros triaxiais foram usados para capturar as componentes ortogonais da aceleração. Determinou-se que apenas 10 cavalos mostraram um padrão de marcha normal, e houve uma correspondência de 43% entre pareceres dos peritos e as técnicas de diagnóstico. De acordo com a classificação das fases da marcha de Stashak, as fases do ciclo da extremidade anterior foram 34/4/8/13/41 enquanto para a extremidade posterior foram 54/11/8/8/19. A amplitude de movimento (ROM) de pescoço, joelho e sesamóide foram 45,52±5,63°, 196,04±19,7° e 209±11,52°, respectivamente. Uma combinação dos métodos experimentais foi utilizada para identificar as fases do ciclo de marcha de cavalos saudáveis. Constatou-se correspondência na localização dos pontos de deslocamento máximo do membro com ambas as técnicas. A informação mais detalhada do movimento dos membros foi obtida usando a acelerometria. Estes métodos são aplicáveis em outras condições, quer a campo ou no laboratório.(AU)

Avaliação de andamentos pelo método acelerométrico Equimetrix em cavalos Mangalarga Marchador

O método acelerométrico, para a análise biomecânica quando usado a campo tem mostrado vantagens com relação a videogrametria, permitindo obter dados cinemáticos e cinéticos e possuindo um processamento de dados mais rápido. Objetivou-se avaliar o efeito do crescimento sobre as características biomecânicas do passo e da marcha de potros da raça Mangalarga Marchador a partir dos 2 meses até três anos e meio de idade, comparando-os com adultos da mesma raça, e estabelecer parâmetros de referência para a raça. Os potros foram equipados com o sistema de análise de andamento Equimetrix® e acompanharam a trajetória da mãe, guiada no cabresto pelo treinador, a marcha e ao passo, com velocidade acompanhada por GPS. As variáveis cinemáticas (comprimento e frequência das passadas, regularidade, simetria e deslocamento dorsoventral do animal) e cinética (propulsão, potências dorsoventral e mediolateral e potência total) foram calculadas e obtidas com o equipamento de análise de andamento Equimetrix®. ®. Esta técnica grava os dados das acelerações 3D (longitudinal, dorsoventral and mediolateral) do movimento. Durante o passo e à marcha, a maioria das variáveis cinemáticas e cinéticas apresentaram diferenças entre as duas idades, exceto na regularidade ao passo (0) e à marcha (p= 0,4331), simetria ao passo (p= 0,7511) e marcha (p= 0,8863) e o deslocamento dorsoventral ao passo (p= 0,1593). Conclui-se que a maior parte das características biomecânicas (cinemáticas e cinéticas) avaliadas, se alterou nos potros da raça Mangalarga Marchador, à marcha e ao passo, nos primeiros três anos e meio de idade. Para determinar se este padrão será mantido, sugere-se a avaliação da marcha destes potros durante a idade adulta.

The acelerometric method when used to analyze biomechanical parameters in the field has shown advantages in relation to the videogrammetry, enabling to obtain kinematic and kinetic data, and allowing faster gait data processing. The goals of this study was to evaluate the biomechanical characteristics of 150 foals, Mangalarga Marchador breed, at 2 months to three and a half years old, to establish parameters for the march gait in this breed. Foals were equipped with the gait analysis system Equimetrix®, and were allowed to follow the trajectory of their dams, at speed controlled by GPS. Kinematic (length and frequency of steps, and dorsoventral displacement of the animal) and kinetics (propulsion force) variables were calculated and obtained with the Equimetrix®. This technique is helpful to record the 3D acceleration data (longitudinal, dorsoventral and mediolateral) during the exercise. During step and march, most kinematic and kinetic characteristics did change in the foals. During step, mean values for regularity (R), simmetry (S) and dorsoventral displacement (DDV) were the same and in the march, mean values for regularity (R) and symmetry (S) were statistically the same between the ages. It was concluded that most of the biomechanical characteristics (kinematic and kinetic) evaluated, did change in the Mangalarga Marchador foals, gait and step in the first third and a half years of age. To determine if this pattern will be maintained, we suggest to assess the gait of these foals during adulthood.

ANÁLISE CINEMÁTICA DA ARTICULAÇÃO METACARPOFALANGEANA DURANTE A RECEPÇÃO DO SALTO EM EQUINOS

A cinemática do salto apresenta variações morfológicas entre raças. Fatores específicos dos cavalos Brasileiros de Hipismo motivaram uma investigação buscando identificar fatores que podem influenciar no desempenho e capacidade atlética de animais desta raça. O objetivo foi avaliar, por meio de análise cinemática, a variação do ângulo metacarpofalangeano, durante a recepção do salto em diferentes alturas ou larguras. Para tanto, avaliaram-se 14 equinos da raça Brasileiro de Hipismo com marcadores reflexivos na face lateral esquerda da parede do casco dos membros torácicos, na articulação metacarpofalangeana e no carpo. Foram avaliados saltos verticais de 90 e 120 cm de altura (V 90 e V 120), e saltos do tipo oxer nas mesmas alturas (Ox 90 e Ox 120), com 100 cm de largura. As avaliações tomaram como base os ângulos em dois momentos, primeiro com o animal em estação; o segundo, durante a recepção do salto, no momento em que a articulação metacarpofalagenana atingiu sua maior hiperextensão. Os ângulos em estação variaram entre 140,17° e 162,37°, com média 151,35° ± 6,32°. Para o primeiro membro, a variação de percentual entre as médias, na comparação entre estação e saltos foi de 22,53 para Estação e V90; 25,4% para Estação e V 120; 24,1% para Estação e Ox 90; e 26,3% para Estação e Ox 120. Na comparação entre os saltos, a variação percentual foi de 3,7% entre V 90 e V 120; 2% entre V 90 e Ox 90; 4,8% entre V 90 e Ox 120; 1,8% entre V120 e Ox 90; 1,1% entre V 120 e Ox 120; e de 2,9% entre Ox 90 e Ox 120. Já para o segundo membro, a variação de percentual entre as médias foi de 17,36% para Estação e V 90; 18,23% para Estação e V 120; 18,61% para Estação e Ox 90; 19,56% para estação e Ox 120. Já entre os saltos, os valores encontrados foram de 1,05% entre V 90 e V 120; 1,51% entre V 90 e Ox 90; 2,65% entre V 90 e Ox 120; 0,46% entre V 120 e Ox 90; 1,62% entre V 120 e Ox 120; e 1,15% entre Ox 90 e Ox 120. Os resultados demonstraram que com o aumento da complexidade do salto, não houve diferença significativa na variação do ângulo da articulação durante a recepção.

The kinematics of the jump has morphological variations between horse breeds. Specific characteristics of the Brazilian Equestrian horses motivated an investigation seeking to identify the factors that can influence the performance and athletic capacity of animals of this breed. The objective was to evaluate through kinematic analysis the metacarpophalangeal angle variation during the reception of the jump at different heights or widths. Fourteen Brazilian Equestrian horses were evaluated with reflective markers on the left lateral side of the hoof of the thoracic limbs, in the fetlock joint and the carpus. Vertical jumps of 90 and 120 cm in height (V 90 and V 120), and oxer-type jumps at the same heights (Ox 90 and Ox 120), 100 cm wide, were evaluated. The assessments were based on joint angles in two moments, first with the animal in standing position; and second during the reception of the jump, at the moment when the fetlock joint reached its greatest hyperextension. The standing position angles varied between 140,17° and 162,37°, with an average of 151,35 ± 6,32°. For the first limb, the percentage variation between the averages, comparing standing position and jumping, was: 22,53% for Standing and V90; 25,4% for Standing and V120; 24,1% for Standing and Ox 90; and 26,3% for Standing and Ox 120. In the comparison between jumps, the percentage variation was 3,7% between V90 and V120; 2% between V90 and Ox90; 4,8% between V90 and Ox120; 1,8% between V120 and Ox90; 1.1% between V120 and Ox120; and 2,9% between Ox90 and Ox120. For the second limb, the percentage variation between the averages was 17,36% for Standing and V90; 18,23% for Standing and V120; 18,61% for Standing and Ox90; 19.56% for Standing and Ox120. Among the jumps, the values found were 1,05% between V90 and V120; 1,51% between V90 and Ox90); 2,65% between V90 and Ox120; 0.46% between V120 and Ox90; 1,62% between V120 and Ox120; and 1,15% between Ox90 and Ox120. The results showed that, with the increase in the complexity of the jump, there was no significant difference in the variation of the angle of the joint during reception.

Kinematic and spermatic recovery after selection by centrifugation in colloid solutions of ovine cryopreserved semen / Cinética e recuperação espermática após seleção por centrifugação em soluções coloidais de sêmen ovino criopreservado

Frozen and thawed ovine semen undergo morphological and functional changes that prevent or decrease the efficiency of fertilization. Sperm selection methods seek to improve the quality and viability of the fertilizing materials. Four sperm selection methods were employed, using two silica colloidal solutions coated with silane (silica colloidal-silane) or by polyvinylpyrrolidone (silica colloidal-PVP), and varying the volume of colloidal solution. Sperm kinematic and sperm recovery were evaluated by means of CASA. The protocols using silica colloidal-silane showed higher total motility (TM), progressive motility (PM) and percentage of rapid sperm (%RAP) compared to the methods employing silica colloidal-PVP and to the samples prior to sperm selection. The silica colloidal-PVP had greater sperm recovery compared to the silica colloidal-silane. Only the method using 4mL of silica colloidal-PVP was not efficient in selecting samples with better quality compared to the samples analyzed prior to sperm selection. The methods using lower volumes of colloidal solution did not differ from those using higher volumes and the best results were shown by the method with 1mL silica colloidal-silane. The results found in the study indicated greater efficiency of the silica colloidal-silane solution for sperm selection of thawed ovine semen when compared to selection using silica colloidal-PVP. The method using 1mL of silica colloidal-silane was equally efficient to the method with higher volume, presenting itself as an alternative to process samples with lower sperm concentration.(AU)

O sêmen ovino congelado e descongelado sofre alterações morfofuncionais que impossibilitam ou diminuem a eficiência na fecundação. Os métodos de seleção espermática visam melhorar a qualidade e a viabilidade do material fecundante. Foram utilizados quatro métodos de seleção espermática utilizando duas soluções de sílica coloidal revestida por silano (sílica coloidal-silano) ou por polivinilpirrolidona (sílica coloidal-PVP), variando o volume de solução coloidal. Foram testadas a cinética espermática no CASA e a recuperação espermática. Os protocolos utilizando sílica coloidal-silano apresentaram maior motilidade total (MT), motilidade progressiva (MP) e porcentagem de espermatozoides rápidos (% RAP) quando comparados aos métodos utilizando a sílica coloidal-PVP e às amostras antes da seleção espermática. A sílica coloidal-PVP teve maior recuperação espermática quando comparada à sílica coloidal-silano. Somente o método utilizando 4mL de sílica coloidal-PVP não foi eficiente na seleção de amostras com melhor qualidade quando comparado às amostras analisadas antes da seleção espermática. Os métodos utilizando menores volumes de solução coloidal não diferiram dos métodos de maior volume, sendo a sílica coloidal-silano com 1mL o método que apresentou os melhores resultados. Como conclusão, os resultados encontrados no trabalho apontaram a maior eficiência da sílica coloidal-silano em selecionar sêmen ovino congelado e descongelado quando comparado à seleção em sílica coloidal-PVP. O método utilizando 1mL de sílica coloidal-silano foi igualmente eficiente ao método com maior volume, sendo uma alternativa para processar amostras com baixa concentração espermática.(AU)

Kinematic and spermatic recovery after selection by centrifugation in colloid solutions of ovine cryopreserved semen / Cinética e recuperação espermática após seleção por centrifugação em soluções coloidais de sêmen ovino criopreservado

Frozen and thawed ovine semen undergo morphological and functional changes that prevent or decrease the efficiency of fertilization. Sperm selection methods seek to improve the quality and viability of the fertilizing materials. Four sperm selection methods were employed, using two silica colloidal solutions coated with silane (silica colloidal-silane) or by polyvinylpyrrolidone (silica colloidal-PVP), and varying the volume of colloidal solution. Sperm kinematic and sperm recovery were evaluated by means of CASA. The protocols using silica colloidal-silane showed higher total motility (TM), progressive motility (PM) and percentage of rapid sperm (%RAP) compared to the methods employing silica colloidal-PVP and to the samples prior to sperm selection. The silica colloidal-PVP had greater sperm recovery compared to the silica colloidal-silane. Only the method using 4mL of silica colloidal-PVP was not efficient in selecting samples with better quality compared to the samples analyzed prior to sperm selection. The methods using lower volumes of colloidal solution did not differ from those using higher volumes and the best results were shown by the method with 1mL silica colloidal-silane. The results found in the study indicated greater efficiency of the silica colloidal-silane solution for sperm selection of thawed ovine semen when compared to selection using silica colloidal-PVP. The method using 1mL of silica colloidal-silane was equally efficient to the method with higher volume, presenting itself as an alternative to process samples with lower sperm concentration.(AU)

O sêmen ovino congelado e descongelado sofre alterações morfofuncionais que impossibilitam ou diminuem a eficiência na fecundação. Os métodos de seleção espermática visam melhorar a qualidade e a viabilidade do material fecundante. Foram utilizados quatro métodos de seleção espermática utilizando duas soluções de sílica coloidal revestida por silano (sílica coloidal-silano) ou por polivinilpirrolidona (sílica coloidal-PVP), variando o volume de solução coloidal. Foram testadas a cinética espermática no CASA e a recuperação espermática. Os protocolos utilizando sílica coloidal-silano apresentaram maior motilidade total (MT), motilidade progressiva (MP) e porcentagem de espermatozoides rápidos (% RAP) quando comparados aos métodos utilizando a sílica coloidal-PVP e às amostras antes da seleção espermática. A sílica coloidal-PVP teve maior recuperação espermática quando comparada à sílica coloidal-silano. Somente o método utilizando 4mL de sílica coloidal-PVP não foi eficiente na seleção de amostras com melhor qualidade quando comparado às amostras analisadas antes da seleção espermática. Os métodos utilizando menores volumes de solução coloidal não diferiram dos métodos de maior volume, sendo a sílica coloidal-silano com 1mL o método que apresentou os melhores resultados. Como conclusão, os resultados encontrados no trabalho apontaram a maior eficiência da sílica coloidal-silano em selecionar sêmen ovino congelado e descongelado quando comparado à seleção em sílica coloidal-PVP. O método utilizando 1mL de sílica coloidal-silano foi igualmente eficiente ao método com maior volume, sendo uma alternativa para processar amostras com baixa concentração espermática.(AU)

Influence of catalase and pre-freezing equilibration on post-thaw semen quality and conception rate in ewes laparoscopically inseminated

The aim of this study was to investigate the effects of catalase and pre-freezing equilibration during ram sperm cryopreservation on motility and membrane and acrosomal integrity of frozen-thawed semen, as well as conception rate following laparoscopic timedinsemination. Semen was collected from four mature Dorper rams, pooled and diluted in Tris egg-yolk extender basic solution (CON), or this solution supplemented with catalase (CAT; 20 U/100 × 106 sperm). Extended semen was packaged in 0.25 ml mini straws (25 × 106 sperm/straw), chilled (to 5°C), and then either frozen immediately (CON and CAT) or maintained at 5°C for 12 h of pre-freezing equilibration (CON12 and CAT12). Immediately after thawing and at 1 h after incubation at 37°C, kinematic parameters (CASA), plasma membrane integrity (PI-FITC), and acrosomal status (FITC-PNA) of sperm were assessed. There were no significant differences among the four groups on sperm traits evaluated immediately postthaw. However, after 1 h of incubation, total motility (46.7 and 25.0%) and plasma membrane integrity (38.7 and 25.7%) were higher (P < 0.05) in CAT12 than CON. When these two treatments were used for laparoscopic timed artificial insemination of ewes (with synchronized ovulation), conception rate was similar for CAT12 and CON (32.8%, n = 61 vs. 27.3%, n = 55). In conclusion, the combination of catalase and pre-freezing equilibration resulted in significantly improved quality of post-thawed ram semen without affecting conception rate in fixed-time laparoscopically intrauterine inseminated ewes.

Influence of catalase and pre-freezing equilibration on post-thaw semen quality and conception rate in ewes laparoscopically inseminated

The aim of this study was to investigate the effects of catalase and pre-freezing equilibration during ram sperm cryopreservation on motility and membrane and acrosomal integrity of frozen-thawed semen, as well as conception rate following laparoscopic timedinsemination. Semen was collected from four mature Dorper rams, pooled and diluted in Tris egg-yolk extender basic solution (CON), or this solution supplemented with catalase (CAT; 20 U/100 × 106 sperm). Extended semen was packaged in 0.25 ml mini straws (25 × 106 sperm/straw), chilled (to 5°C), and then either frozen immediately (CON and CAT) or maintained at 5°C for 12 h of pre-freezing equilibration (CON12 and CAT12). Immediately after thawing and at 1 h after incubation at 37°C, kinematic parameters (CASA), plasma membrane integrity (PI-FITC), and acrosomal status (FITC-PNA) of sperm were assessed. There were no significant differences among the four groups on sperm traits evaluated immediately postthaw. However, after 1 h of incubation, total motility (46.7 and 25.0%) and plasma membrane integrity (38.7 and 25.7%) were higher (P < 0.05) in CAT12 than CON. When these two treatments were used for laparoscopic timed artificial insemination of ewes (with synchronized ovulation), conception rate was similar for CAT12 and CON (32.8%, n = 61 vs. 27.3%, n = 55). In conclusion, the combination of catalase and pre-freezing equilibration resulted in significantly improved quality of post-thawed ram semen without affecting conception rate in fixed-time laparoscopically intrauterine inseminated ewes.(AU)

Controlled traffic and soil physical quality of an Oxisol under sugarcane cultivation

Machinery traffic in sugarcane (Saccharumsp.) plantations reduces soil physical quality, and hinders both root development and crop yield. We evaluated the physical quality of an Oxisol and the development of sugarcane roots under controlled traffic. The treatments assessed were: without controlled machinery traffic (WCT), controlled traffic by adjusting the tractor and infield wagons to a 3.0 m track width with the operator guiding the machinery (CT1) and the previous treatment using real time kinematic / global positioning system (RTK / GPS) precision auto steer (CT2). Soil samples were collected from the planting rows, seedbed and inter-row center to determine the least limiting water range (LLWR) and soil porosity from scanned 2-D images. The root dry mass was sampled from monoliths, separated from the soil by washing through a 2-mm sieve and dried in an oven. A higher LLWR was observed in the planting row under CT1 and CT2 than under WCT. The planting row had a predominance of complex pores with a diameter > 500 µm in the 0.15-0.27 m depth layer under CT1 and CT2. In the planting rows under WCT, the root dry mass was only 44 % of that measured under CT2. Benefits regarding soil physical quality and growth roots were observed when the tractor-wagon track width was adjusted based on the sugarcane spacing using either precision auto steering or manual operation of the machinery.

Controlled traffic and soil physical quality of an Oxisol under sugarcane cultivation

Machinery traffic in sugarcane (Saccharumsp.) plantations reduces soil physical quality, and hinders both root development and crop yield. We evaluated the physical quality of an Oxisol and the development of sugarcane roots under controlled traffic. The treatments assessed were: without controlled machinery traffic (WCT), controlled traffic by adjusting the tractor and infield wagons to a 3.0 m track width with the operator guiding the machinery (CT1) and the previous treatment using real time kinematic / global positioning system (RTK / GPS) precision auto steer (CT2). Soil samples were collected from the planting rows, seedbed and inter-row center to determine the least limiting water range (LLWR) and soil porosity from scanned 2-D images. The root dry mass was sampled from monoliths, separated from the soil by washing through a 2-mm sieve and dried in an oven. A higher LLWR was observed in the planting row under CT1 and CT2 than under WCT. The planting row had a predominance of complex pores with a diameter > 500 µm in the 0.15-0.27 m depth layer under CT1 and CT2. In the planting rows under WCT, the root dry mass was only 44 % of that measured under CT2. Benefits regarding soil physical quality and growth roots were observed when the tractor-wagon track width was adjusted based on the sugarcane spacing using either precision auto steering or manual operation of the machinery.(AU)

Influência da posição de cabeça e pescoço na cinemática da pelve e articulação coxofemoral de equinos durante a marcha

No treinamento para esportes equestres ocorre a alteração da posição natural de cabeça e do pescoço para uma posição definida pelo cavaleiro. A melhor posição para o treinamento gera debate constante entre os treinadores, pois existem controvérsias sobre os efeitos dessas posições na cinemática dos cavalos. O objetivo deste trabalho é realizar uma análise cinemática tridimensional da pelve e da articulação coxofemoral de cavalos Mangalarga Marchador de marcha batida durante a marcha, e a influência da posição da cabeça e do pescoço, no padrão do movimento angular e na amplitude média de movimento. Além disso, relatar dados cinemáticos não publicados de cavalos Mangalarga Marchador para entender a marcha batida. Para análise cinemática tridimensional, foi utilizado um sistema optoeletrônico. Dezenove câmeras OptiTrack Prime 17W (360 Hz) foram posicionadas formando um volume de aquisição de 16 × 4,8 × 3 metros. Dez marcadores retrorrefletivos foram fixados bilateralmente na tuberosidade coxal e sacral, no fêmur no trocânter maior, terceiro trocânter e epicôndilo lateral. Cada animal realizou três tentativas na marcha, com no mínimo três passadas completas, nas 4 posições de cabeça e pescoço avaliadas. As coordenadas tridimensionais dos marcadores foram filtradas com filtro Butterworth de segunda ordem e passa-baixo. Foram calculados os ângulos entre a pelve e o segmento lombar da coluna e da articulação coxofemoral a partir de sistemas de coordenadas locais. No movimento de flexão e extensão, a posição Livre apresentou 2,28º de amplitude, com aumento (P <0,05) para 2,72 na posição Vertical. A flexão lateral mostrou redução (P <0,05) na posição vertical, com 1,71º de amplitude. Na articulação coxofemoral, houve redução (P <0,05) na amplitude de flexão e extensão nas posições Invertido e Vertical em relação à posição Livre. A amplitude de adução e abdução aumentou (P <0,05) na articulação coxofemoral esquerda e a rotação diminuiu (P <0,05) na direita na posição invertida. Durante o ciclo da passada a os valores médios do ângulo da pelve e da articulação coxofemoral foram maiores no eixo x do que as demais posições, indicando aumento da inclinação da pelve e do fêmur

Equestrian sports involves training of the horse with the head and neck placed in a position defined by the rider. The best position for training is currently under debate among riders and trainers, but there are few scientific data available to confirm or disprove the different views. There is controversy about the effects of these positions on kinematics of horses, but few quantitative data are available. To provide a analise of 3D kinematics (movements) of the pelvis and hip joint of Mangalarga Marchador horses during the marcha batida gait and the influence of head and neck position, on the pattern of angular movement, and a medium amplitude of movement. In addition, to report unpublished kinematic data of Mangalarga Marchador horses to understand the marcha batida gait. For three-dimensional kinematic analysis, an optoelectronic system was used. Nineteen OptiTrack Prime 17W cameras (360 Hz) were positioned around the animals, and an acquisition volume of 16 × 4.8 × 3 meters was used. Ten retroreflective markers were fixed bilateral on the coxal and sacral tubrerosity, on the femur at cranial greater trochanter, third trochanter and lateral epicondyle. Each animal performed three trials at marcha batida gait with three complete gait cycles each with the head and neck in 4 different head and neck positions. The 3D coordinates of the markers were filtered with a second-order, low-pass, Butterworth filter. We obtained joint angles between the pelvis and the lumbar back segment and the hip joint. In the flexion and extension movement, the Free position presented 2.28º of amplitude, with an increase (P <0.05) to 2.72 in the Vertical position. The lateral flexion showed a reduction (P <0.05) in the Vertical position, with 1.71º of amplitude. In the hip joint, there was a reduction (P <0.05) in the amplitude of flexion and extension in the Inverted and Vertical positions in relation to the Free position. The amplitude of adduction and abduction increased (P <0.05) in the Left hip and the rotation decreased (P <0.05) in the Right hip in the Inverted position. During the stride cycle, the pelvis was more flexed and the hip was more extended in the inverted position. The vertical position increased the range of motion of pelvis flexion and extension. The Inverse position increases the inclination of the pelvis and femur throughout the stride cycle.