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)

Comparison of maximal lactate steady state with V2, V4, individual anaerobic threshold and lactate minimum speed in horses / Comparação da máxima fase estável do lactato com a V2, V4, o limiar anaeróbio individual e lactato mínimo em equinos

The anaerobic threshold is a physiologic event studied in various species. There are various methods for its assessment, recognized in the human and equine exercise physiology literature, several of these involving the relationship between blood lactate concentration (LAC) and exercise load, measured in a standardized exercise test. The aim of this study was to compare four of these methods: V2, V4, individual anaerobic threshold (IAT) and lactate minimum speed (LMS) with the method recognized as the gold standard for the assessment of anaerobic threshold, maximal lactate steady-state (MLSS). The five tests were carried out in thirteen trained Arabian horses, in which velocities and associated LAC could be measured. The mean velocities and the LAC associated with the anaerobic threshold for the five methods were respectively: V2 = 9.67±0.54; V4 = 10.98±0.47; V IAT = 9.81±0.72; V LMS = 7.50±0.57 and V MLSS = 6.14±0.45m.s-1 and LAC IAT = 2.17±0.93; LAC LMS = 1.17±0.62 and LAC MLSS = 0.84±0.21mmol.L-1. None of the velocities were statistically equivalent to V MLSS (P<0.05). V2, V4 and V LMS showed a good correlation with V MLSS , respectively: r = 0.74; r = 0.78 and r = 0.83, and V IAT did not significantly correlate with V MLSS. Concordance between the protocols was relatively poor, i.e., 3.28±1.00, 4.84±0.30 and 1.43±0.32m.s-1 in terms of bias and 95% agreement limits for V2, V4 and LMS methods when compared to MLSS. Only LAC LMS did not differ statistically from LAC MLSS. Various authors have reported the possibility of the assessment of anaerobic threshold using rapid protocols such as V4 and LMS for humans and horses. This study corroborates the use of these tests, but reveals that adjustments in the protocols are necessary to obtain a better concordance between the tests and the MLSS.

O limiar anaeróbio é um evento fisiológico estudado em várias espécies. Sua mensuração possui vários métodos reconhecidos na literatura da fisiologia do exercício humano e equino, muitos deles envolvendo a relação entre a concentração sanguínea de lactato (LAC) e a carga de exercício. O objetivo do presente estudo foi comparar quatro desses métodos: V2 , V4 , limiar anaeróbio individual (LAI) e o teste do lactato mínino (LM) com o método reconhecido na literatura como o padrão ouro para a mensuração do limiar anaeróbio, a máxima fase estável do lactato (MFEL). Os cinco testes foram realizados em treze equinos árabes treinados, nos quais as velocidades e suas respectivas LAC puderam ser quantificadas. As velocidades médias e LAC associadas ao limiar anaeróbio aferido pelos cinco métodos foram respectivamente: V2 = 9,67±0,54; V4 = 10,98±0,47; V LAI = 9.81±0.72; V LM = 7,50±0,57 e V MFEL = 6,14±0,45m.s-1 ; e LAC LAI = 2,17±0,93; LAC LM = 1,17±0,62 e LAC MFEL = 0,84±0,21mmol.L-1. Nenhuma dessas velocidades foi estatisticamente igual à V MFEL (P<0,05). A V2 , a V4 e a V LM mostraram uma boa correlação com a V MFEL , respectivamente r = 0,74; r = 0,78 e r = 0,83, e a V LAI não se correlacionou significativamente com a V MFEL. A concordância entre os protocolos foi relativamente fraca, sendo 3,28±1,00; 4,84±0,30 e 1,43±0,32m.s-1 em termos de viés e limites de concordância a 95% para os métodos V2 , V4 e LM comparados à MFEL. Muitos autores relataram a possibilidade da mensuração do limiar anaeróbio pelo uso de protocolos rápidos, como a V4 e o LM, para humanos e equinos. O presente estudo corrobora a utilização desses testes, mas revela que ajustes nos protocolos são necessários para se obter uma melhor concordância entre os mesmos e a MFEL.

Comparison of the lactate minimum speed and the maximal lactate steady state to determine aerobic capacity in purebred Arabian horses.

AIM: To compare five different protocols for estimating the lactate minimum speed (LMS) with that for estimating the maximal lactate steady state (MLSS) in Arabian horses, in order to obtain a more rapid method for monitoring aerobic capacity and prescribing training schedules. METHODS: Eight purebred Arabian horses were conditioned to exercise on a treadmill for 12 days then submitted to three to five exercise sessions to determine the MLSS. Blood samples were collected from a jugular catheter at specific intervals for measurement of lactate concentrations. The MLSS was the velocity maintained during the last 20 minutes of constant submaximal exercise, at which the concentration of lactate increased by no more than 1.0 mmol/L. The LMS test protocols (P1 - P5) included a warm-up period followed by a high-intensity gallop. The speed was then reduced to 4 m/s, and the incremental portion of the test was initiated. In P1, P2, and P3, the velocity increment was 0.5 m/s, and the duration of each incremental stage was three, five and seven minutes, respectively. In P4 and P5, the velocity increments were 1.0 and 1.5 m/s, respectively, and the duration of the stages was fixed at five minutes each. A second-degree polynomial function was fitted to the lactate-velocity curve, and the velocity corresponding to the lowest concentration of lactate was the LMS. RESULTS: Only the mean LMS determined by P1 and P2 did not differ from the velocity determined by the MLSS test (p > 0.1). There was a strong correlation (r >0.6) between P1 and the MLSS velocity. A limits of agreement plot revealed that the best agreement occurred between the MLSS test and P1 (mean bias = 0.14 m/s), followed by P2 (bias = -0.22 m/s). The lactate concentrations associated with the various LMS protocols did not differ. CONCLUSIONS: This study shows the variation between protocols of the LMS test for determining the onset of blood lactate accumulation but also reveals that, at least for Arabian horses, the P1 protocol of the LMS has good agreement with the MLSS.

Descrição do movimento da articulação metacarpofalangiana de equinos pela metodologia baseada em videogrametria / Equine metacarpophalangeal joint movement using videogrammetry method

Caracterizou-se, mediante análise cinemática tridimensional baseada na videogrametria, o ângulo dorsal da articulação metacarpofalangiana de equinos em sete equinos da raça Puro Sangue Árabe. A análise tridimensional do movimento foi realizada em esteira rolante. O programa Dvideo foi utilizado para a obtenção das coordenadas tridimensionais do sistema de calibração e dos marcadores reflexivos posicionados na extremidade proximal do terceiro osso metacárpico, articulação metacarpofalangiana e extremidade distal da primeira falange. A articulação metacarpofalangiana apresentou extensão máxima durante o momento de apoio, no qual a face lateral do terceiro osso metacarpiano se apresentava de forma perpendicular ao solo. Foram observados dois picos de flexão durante a fase de elevação. Concluiu-se que a instrumentação utilizada para a análise cinemática tridimensional permitiu a investigação quantitativa da variação angular do movimento de extensão e flexão da articulação metacarpofalangiana de equinos por meio de imagens digitalizadas.

The kinematic pattern of the metacarpophalangeal joint of equine using videogrammetry was evaluated. The three dimensional kinematic analysis were performed in a treadmill using seven Arabian horses. The Dvideo program was used to obtain the three dimensional coordinates from a calibration system and three reflective markers placed on the third metacarpus, metacarpophalangeal joint and proximal phalanx of the left forelimb. After the landing of the hoof, the fetlock extends to maximal loading at mid stance. During the swing phase the joint shows two flexion peaks. This method allows the determination of the flexion and extension angle of the metacarpophalangeal joint during locomotion.

The workload and plasma ion concentration in a training match session of high-goal (elite) polo ponies.

REASONS FOR PERFORMING STUDY: This study was designed to consider the complexity of the physical effort inherent to horses in polo competitions and the absence of reports in the literature on the effort, intensity and electrolyte changes resulting from a collective team training session aimed at preparing for a polo championship. OBJECTIVES: To determine the effort and ion changes caused by an outdoor polo training match for a 25 goal handicap (elite) based on physiological variables including acid-base status (venous pH, PCO(2) and HCO(3)(-)), packed cell volume (PCV), haemoglobin (Hb), lactate, glucose, sodium, chloride and potassium and strong ion difference (SID) as well as creatine kinase (CK) activity. METHODS: Twenty-three clinically healthy 'high-goal' polo ponies were used, which included 10 geldings and 13 females. The horses performed a training match, as a preparation for a 25 goal tournament, consisting of 6 chukkas of 7 min duration each. Blood samples were collected during resting, and at 5 min, 6 and 12 h after each chukka. Data were analysed using ANOVA for repeated measures followed by Tukey's test. RESULTS: Differences (P < 0.001) were evident mainly in post exercise for all variables studied. There was a reduction in pH, PCO(2) and HCO(3)(-) and SID, together with an increase in PCV and Hb, lactate, glucose, Na(+) and Cl(-). K(+) levels remained constant at all times of collection. The average resting value for CK was 255 ± 9 iu/l, and 6 h after effort there was a 35% increase in enzyme activity. CONCLUSIONS: This study indicates that the horses participating in a training match underwent a high-intensity effort with alterations in electrolytes and acid-base equilibrium. POTENTIAL RELEVANCE: Training matches should be carefully conducted, with a suitable recovery period before the main match.

Influência do treinamento aeróbio sobre o cortisol e glicose plasmáticos em equinos / Influence of the aerobic training on cortisol and glucose levels in horses

Estudou-se a resposta do cortisol e da glicemia em 12 equinos da raça Puro Sangue Árabe destreinados (T0) por oito meses e submetidos a um período de 90 dias de treinamento aeróbio (T90). Para avaliação dos efeitos do treinamento, empregou-se teste ergométrico constituído de exercício progressivo em esteira rolante, acompanhado por colheitas de sangue 15 segundos antes do término de cada etapa de esforço. A velocidade (intensidade) do treino foi definida como sendo 80 por cento da V4 (velocidade na qual a lactacidemia atinge 4mmol/L). Adicionalmente, no último mês de treinamento, foi instituído, uma vez por semana, exercício com velocidades variáveis, chamado "fartlek". Após 90 dias de treinamento, a concentração plasmática de cortisol elevou-se e após o teste de esforço (20min), houve aumento da glicemia. Este resultado reflete a possibilidade de adaptação ao treinamento. Conclui-se que o cortisol plasmático pode ser utilizado como ferramenta na avaliação de um programa de treinamento em equinos.

Cortisol and glucose responses were evaluated in 12 Arabian (PSA) horses submitted to a detraining period of eight months (T0) and to 90 days of aerobic training (T90). For the evaluation of the effect of training, a standardized incremental exercise test in a treadmill was used. Fifteen seconds before the ending of each effort step, blood samples were collected. The speed (intensity) of the training was defined as being 80 percent of the V4 (speed at which the blood lactate concentration reaches 4mmol/L). Additionally, in the last month of training, velocity play, a type of exercise with varying velocities called "fartlek" was instituted, once a week. Results showed that after 90 days of training, the plasmatic concentrations of cortisol and glucose increased when compared to the untrained horses. This result reflects the possibility of adaptation to the training. The blood cortisol levels may be used as a tool for the evaluation of a training program in horses.

Determinação de eletrólitos, gases sanguíneos, osmolalidade, hematócrito, hemoglobina, base titulável e anion gap no sangue venoso de equinos destreinados submetidos a exercício máximo e submáximo em esteira rolante / Determination of electrolytes, hemogasometry, osmalility, hematocrit, hemoglobin, base concentration, and anion gap in detrained equines submitted a maximum and submaximum exercise on treadmill

Estudaram-se as alterações nos eletrólitos, nos gases sanguíneos, na osmolalidade, no hematócrito, na hemoglobina, nas bases tituláveis e no anion gap no sangue venoso de 11 equinos da raça Puro Sangue Árabe, destreinados, submetidos a exercício máximo e submáximo em esteira rolante. Esses animais passaram por período de três dias de adaptação à esteira rolante e posteriormente realizaram dois exercícios testes, um de curta e outro de longa duração. Foram coletadas amostras de sangue venoso antes, imediatamente após e 30 minutos após o término dos exercícios. Após a realização do exercício máximo, observou-se diminuição significativa no pHv, na PvCO2, no HCO3, na cBase além de elevação no AG. Detectou-se também aumento do K+, do Ht e da Hb. Ao final do exercício submáximo, constatou-se somente aumento significativo no pHv, na cBase, na SatvO2 e na PvO2. Conclui-se que os equinos submetidos a exercício máximo desenvolveram acidose metabólica e alcalose respiratória compensatória, hipercalemia e aumento nos valores de hematócrito e hemoglobina. No exercício submáximo, os animais apresentaram alcalose metabólica hipoclorêmica e não ocorreram alterações no equilíbrio hidroeletrolítico.

Changes in electrolytes, blood gas, osmolality, hematocrit, hemoglobin, base concentration, and anion gap in 11 detrained Arabian horses during exercise on a high-speed treadmill were investigated. After a period of three days of adaptation on the rolling mat, the animals were submitted to two exercises: one of short (maximum) and other of long duration (submaximum). Venous blood samples were obtained right before, and 30 minutes after the exercise. After the maximum exercise, it was observed a significative decrease in pHv, PvCO2, HCO3, and cBase and an increase in AG. It was also observed hypercalemia and increase in Ht and Hb. At the final of the submaximum exercise, it was observed significative increase in pH, cBase, SatvO2, and PvO2. So, maximum exercises can lead equines to present metabolic acidosis with respiratory alkalosis as response, hypercalemia and increase in hematocrit and hemoglobin, values. Submaximum exercises can present hypochloremic metabolic alkalosis but no alterations in the hydroelectrolitic balance.

Alterations in muscular enzymes of horses competing long-distance endurance rides under tropical climate / Alterações em enzimas musculares de cavalos em competições de enduro de longa distância em clima tropical

This study tried to monitor the alterations of muscle enzymes activity - creatinokinase (CK), lactate dehydrogenase (LDH), and aspartate aminotransferase (AST) - in a group of horses that participated of 70 and 100km distance rides in five competitions of an annual endurance championship under tropical climate. Pre ride levels (U/l) were 245.13±9.84 for CK, 496.61±14.76 for LDH, and 328.95±8.65 for AST. When compared to these levels, the results revealed a significant decrease in all enzymes activities in the first moment of the rides. Peak levels, significantly different, were reached, immediately after rides by CK (413.591±50.75); 24 hours post rides by LDH (628.61±33.30); and 48 hours after rides by AST (389.89±16.96). Monitoring of recovery period revealed different behavior among enzymes activities with CK values returning to pre ride values (279.61±23.05) 24 hours post rides, while LDH and AST values returned to pre ride values (505.25±33.78 and 359.35±24.90, respectively) 72 hours post rides. These data revealed different alterations in concentration of muscular enzymes in endurance horses directly related to the duration of the effort.

Estudaram-se as alterações de atividade das enzimas musculares creatino quinase (CK), lactato desidrogenase (LDH) e aspartato aminotransferase (AST) em um grupo de cavalos que utilizados em provas de enduro de 70 e 100km de distância, em cinco competições. Os valores (U/l) basais (antes da largada) foram 245,13±9,84 para CK, 496,61±14,76 para LDH e 328,95±8,65 para AST. Todas as atividades das enzimas decresceram no primeiro momento das provas (~30km). Valores de pico, significativamente diferentes, foram alcançados para CK (413,59±50,75) imediatamente após 70km de distância; 24 horas após para LDH (628,61±33,30); e 48 horas após as provas para AST (389,89±16,96). A monitoração do período de recuperação revelou diferente comportamento entre as concentrações enzimáticas com CK retornando aos valores basais 24 horas pós-provas (279,61 ± 23,05). LDH e AST retornaram aos valores basais, 72 horas pós-provas (505,25±33,78 e 359,35±24,90, respectivamente). Os dados obtidos revelaram diferentes alterações na concentração de enzimas musculares de cavalos de enduro, diretamente relacionadas com a duração do esforço.

Blood lactate threshold reflects glucose responses in horses submitted to incremental exercise test / Limiar sangüíneo de lactato reflete a resposta da glicose em cavalos submetidos a teste de exercício progressivo

Determinou-se, em eqüinos, o efeito do treinamento sobre as concentrações sangüíneas de lactato e plasmáticas de glicose durante exercício de intensidade progressiva em esteira rolante. Demonstrou-se que o treinamento aeróbico causou diminuição da concentração máxima de lactato e que o limiar de lactato corresponde ao ponto de inflexão da curva de glicose plasmática, confirmando esse parâmetro como indicador da capacidade aeróbica de cavalos.

Análise cinemática tridimensional do movimento de eqüinos em esteira rolante / Three-dimensional cinematic analysis of the horse movement in treadmill

Adaptou-se uma metodologia utilizada no estudo do movimento humano para analisar o movimento de eqüinos em esteira rolante de alto desempenho, construindo-se um sistema de calibração utilizando-se um teodolito eletrônico de alta precisão e calibradores que demarcavam o espaço ocupado pelo animal. Dessa forma, foram obtidas as coordenadas espaciais dos marcadores presentes nos calibradores, compondo o arquivo de calibração. Duas câmeras de vídeo (60Hz) foram posicionadas em vista lateral esquerda, distantes 5m da manta de rolagem da esteira a 1,8m de altura. As imagens foram enquadradas utilizando-se um monitor de vídeo. Para a validação do método, um membro da equipe movimentou, no espaço calibrado, uma haste rígida com marcadores fixados nas suas extremidades, distantes um do outro 43,8cm. Para a análise das imagens, utilizou-se o programa Dvideow, desenvolvido pelo Laboratório de Instrumentação para Biomecânica - UNICAMP. Após a reconstrução tridimensional das imagens utilizou-se um programa matemático para o cálculo do valor médio da distância entre os marcadores. Obteve-se, após a mensuração em 100 quadros, o valor médio de 43,7cm com coeficiente de variação de 0,8 por cento. Estes resultados mostram que a metodologia desenvolvida é precisa e adequada para o estudo da locomoção de eqüinos

A methodology used for the study of human movement was adapted to analyze the movement of equines in a high performance treadmill. An electronic high precision theodolite was used to construct a calibration system to demarcate the volume occupied by the animal. It was taken the three dimensions coordinates of each marker present in the calibrator, composing the calibration archive. Two video cameras (60 Hz) were located approximately 5m from the left lateral side of the treadmill at 1.8m height. The images had been fit using a video monitor. For the validation of the method, a member of the team moved into the calibrated space, a 43.8cm long rigid stick with markers settled in its extremities. For the analysis of the images, the Dvideow program developed by the Laboratório de Instrumentação para Biomecânica - UNICAMP was used. After the three-dimensional reconstruction of the images, the calculation of the distance between the markers of the stick was done in 100 frames. The mean was 43.7 cm with a variation coefficient of 0.8 percent. These results confirm that this methodology is precise and adequate for the study of the locomotion in horses

Sedative and antinociceptive effects of romifidine and xylazine in Thoroughbred mares

The sedative and antinociceptive effects of romifidine (0.1mg/kg) and of xylazine (1.0mg/kg) on Thoroughbred mares were studied. Sedation was evaluated by quantifying spontaneous locomotor activity (SLA) and head height (HH) in animals placed in automated individual behaviour stalls. Antinociception was determined utilizing a heat irradiation lamp recording the latency time for the hoof withdrawal reflex (HWRL) and the latency time for the skin twitch reflex (STRL) in a randomised block design with 10 replicates. Comparison of the sedative effects of romifidine and of xylazine on SLA showed a faster effect for xylazine. Regarding the sedative effect of the substances based on HH, romifidine caused a longer-lasting effect. Romifidine caused an increase in HWRL and STRL, and xylazine, an agent known for its analgesic effect, did not have an antinociceptive effect based on STRL result. The antinociceptive effect of romifidine was more pronounced than the xylazine effect.

Compararam-se os efeitos sedativos e antinociceptivos da romifidina (0,1mg/kg) e da xilazina (1,0mg/kg) em éguas da raça Puro Sangue Inglês. A sedação foi quantificada por meio da atividade locomotora espontânea (ALE) e altura da cabeça (AC) em baias individuais automatizadas para o estudo do comportamento. A antinocicepção foi avaliada utilizando uma lâmpada de irradiação de calor registrando-se a latência para o reflexo de retirada do membro (LRRM) e a latência para o reflexo do frêmito cutâneo (LRFC), em delineamento experimental em blocos ao acaso com 10 repetições. O efeito sedativo sobre a ALE foi de aparecimento mais rápido no grupo exposto à xilazina, ao passo que a ptose da cabeça foi mais prolongada no grupo que recebeu romifidina. A romifidina promoveu aumento da LRRM e LRFC e a xilazina não causou efeito antinociceptivo medido pela LRFC. O efeito antinociceptivo da romifidina foi mais pronunciado que o da xilazina.

Composição de fibras musculares esqueléticas de eqüinos jovens da raça Brasileiro de Hipismo / Composition of skeletal muscle fibers of young Brasileiro de Hipismo horse breed

The aim of this study was to typify the skeletal striated fibers of the gluteus medius muscle of young Brasileiro de Hipismo (BH) horses by means of histochemical analysis with m-ATPase and NADH-TR according to the sex and the biopsy depth. It was observed that the frequency (F; percent) and the relative cross sectional area (RCSA; percent) of the fibers type IIX were greater than the fibers type IIA, which F and RCSA were greater than the fibers type I. The comparison between sex and muscles depht, showed no significant difference in F and RCSA in the three types of fibers. The results of morphometry showed that the gluteus medius muscle has greater glycolitic metabolism and anaerobic capacity because of the presence of large proportion of type IIX fibers. This may be justified by the genetic influence of Thoroughbred in the formation of Brasileiro de Hipismo breed.

Aerobic training, but not creatine supplementation, alters the gluteus medius muscle.

The aim of the present study was to investigate the effect of oral supplementation of creatine on the muscular responses to aerobic training. Twelve purebred Arabian horses were submitted to aerobic training for 90 d, with and without creatine supplementation, and evaluated with respect to BW and BCS and to the area and frequency of the different types of muscle fibers in the gluteus medius. Supplementation consisted of the daily administration of 75 g of creatine monohydrate mixed into the ration for the 90 d of training. Physical conditioning was conducted on a high-performance treadmill, and training intensity was stipulated by calculating the velocity at which blood lactate reaches 4 mmol/L, determined monthly for each animal. The individual intensity of physical force at 80% of aerobic threshold was established. Morphometry of gluteus medius muscle fibers was performed on frozen sections processed for histochemical analysis of myosin adenosine triphosphatase and immunohistochemistry of slow-contracting myosin. The results demonstrated that the animals maintained a moderate BCS without alteration of BW during the course of training, providing evidence of equilibrium between food intake and caloric expenditure during the study period. The present study demonstrated that aerobic training for 90 d caused hypertrophy of fiber types I (P = 0.04), IIA (P = 0.04), and IIX (P = 0.01), as well as an increase in the relative area occupied by type I fibers (P = 0.02) at the expense of type IIX fibers (P = 0.03), resulting in modifications of the contractile and metabolic characteristics of the gluteus medius muscle. It was not possible to show any beneficial effect from creatine on the skeletal muscle characteristics examined.

Study of caffeine in urine and saliva of horses subjected to urinary acidification.

The study of caffeine in racing horses has been of growing concern in veterinary sports medicine since the Association of Racing Commissioners International (ARCI) stated that it has no valid therapeutic use in racehorses. We examined the kinetic alterations in the urinary excretion and salivary secretion of caffeine in seven horses subjected to urinary acidification using ascorbic acid because this procedure can simulate the acidosis that follows anaerobic exercise. They participated in two treatment groups: the control group (SG) received 500 ml of saline and then 2.0 mg kg(-1) caffeine i.v. 30 min later; and the acidi fi ed group (AG) was subjected to urinary acidification with ascorbic acid at a dose of 0.5 g kg(-1) i.v. and then 2.0 mg kg(-1) caffeine i.v. 30 min later. Samples were collected 30 min before caffeine administration, immediately before caffeine administration (time zero) and at 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, 48 and 72 h afterwards. The samples were assayed by gas chromatography. The mean urinary pH for SG was 8.2, but for AG it was as low as 5.9 at 4 h, extending acidosis for up to 8 h. The kinetic curves for the two groups were similar for urinary excretion and salivary secretion. Differences occurred only in peak excretion and peak secretion in SG obtained at 1 h and 30 min, respectively, and in AG at 2 h and 1 h, respectively. This could be explained, in part, to the diuresis in AG compared with SG, resulting in less concentrated urine in the former group. The large difference between the pKa of caffeine and the pH of the medium may be responsible for the similar pharmacokinetics observed for the two groups.

Comparative study of the sedative and antinociceptive effects of levomepromazine, azaperone and midazolam in laboratory animals

Os efeitos sedativos e antinociceptivos da levomepromazina, azaperone e midazolam foram avaliados utilizando-se três testes de comportamento em ratos e camundongos. No teste da atividade locomotora espontânea em campo aberto observou-se que tanto o comportamento exploratório como a atividade locomotora espontânea foram significativamente diminuídos quando se utilizou levomepromazina e azaperone. O efeito causado pelo azaperone foi menos prolongado quando comparado ao da levomepromazina. O midazolam causou diminuição do comportamento exploratório sem alterar a atividade locomotora espontânea. Quando se avaliou o efeito antinociceptivo por meio da latência para o reflexo da retirada da cauda em ratos após estímulo doloroso, as drogas não apresentaram nenhum efeito antinociceptivo observável. No teste das contorções em camundongos, os fármacos foram capazes de abolir as contorções quando comparados ao efeito do grupo-controle. Levomepromazina, azaperone e midazolam nas doses utilizadas foram capazes de inibir o comportamento exploratório de ratos, comprovando seus efeitos sedativos. Com relação aos efeitos antinociceptivos para dor visceral, eles foram capazes de inibir as contorções.

Toxicological evaluation of long-term intravenous administration of amitraz in horses

With the aim of determining the possible toxicity of amitraz after its prolonged use in horses, six English Thoroughbred horses received intravenous injections of amitraz (0.05, 0.10 or 0.15 mg/kg) weekly for four months, constituting the experimental group. Eight other animals (control group), via the same route following the same drug administration schedule and period of time, received the vehicle, dimethylformamide. At the end of this period, blood was collected from all the animals, and a comparison was made of the means of the values obtained for the various blood analyses: complete hemogram, alkaline phosphatase, gamma-glutamyltransferase, blood urea nitrogen, lactate dehydrogenase, aspartate aminotransferase, creatine phosphokinase, glucose, albumin, total protein, creatinine, Na+ , K+, Cl- and CO2. The results for the biochemical characteristics showed that only the mean value for urea of the animals submitted to treatment with amitraz was significantly different than the mean value obtained for the control group. The analyses of the hematological characteristics showed that no significant differences between groups were observed. Similarly, the measurement of blood electrolyte levels demonstrated that long-term treatment with amitraz did not cause significant changes in the variables analyzed. The results indicate that amitraz, given in the doses employed in this study, did not show signs of inducing toxic effects in vital organs, even after prolonged administration

Determination of the highest no-effect dose (HNED) and of the elimination pattern for cocaine in horses.

Cocaine is one of the most widespread illegal stimulants utilized by the human population throughout the world. The aim of this study was to establish the highest no-effect dose (HNED) of cocaine on the spontaneous locomotor activity (SLA) of horses in a behavior chamber, and thereby to determine the maximal acceptable threshold of the urinary drug concentration in horses. Twelve English thoroughbred mares received 0.02, 0.03, 0.04, 0.08 or 0.12 mg kg(-1) cocaine i.v. or saline solution (control). It was noted that doses above 0.04 mg kg(-1) induced a significant increase in SLA (P < 0.05, Tukey's test). No significant increase in SLA was seen in the mares that received 0.03 mg kg(-1), but the animals showed important behavioral changes that did not occur after the 0.02 mg kg(-1) dose. It was concluded that the HNED of cocaine for horses in a behavior chamber is 0.02 mg kg(-1). After injection of this dose in five horses, urine samples were collected at predetermined intervals through vesical catheterization. The concentrations of cocaine, norcocaine, benzoylecgonine and ecgonine methyl ester were quantified by liquid chromatography/electrospray ionization tandem mass spectrometry. Cocaine and norcocaine concentrations remained consistently below the level of detection. Benzoylecgonine reached a mean (+/- SEM) maximum concentration of 531.9 +/- 168.7 ng ml(-1) after 4 h, whereas ecgonine methyl ester peaked 2 h after injection at a concentration of 97.2 +/- 26.5 ng ml(-1). The maximum admissible concentration for cocaine and/or metabolites in the urine of horses is difficult to establish unequivocally because of the substantial individual variation in the drug elimination pattern observed in horses, which can be inferred by the large standard error of the means obtained.

Effects of caffeine on locomotor activity of horses: determination of the no-effect threshold.

Caffeine is the legal stimulant consumed most extensively by the human world population and may be found eventually in the urine and/or blood of race horses. The fact that caffeine is in foods led us to determine the highest no-effect dose (HNED) of caffeine on the spontaneous locomotor activity of horses and then to quantify this substance in urine until it disappeared. We built two behavioural stalls equipped with juxtaposed photoelectric sensors that emit infrared beams that divide the stall into nine sectors in a 'tic-tac-toe' fashion. Each time a beam was interrupted by a leg of the horse, a pulse was generated; the pulses were counted at 5-min intervals and stored by a microcomputer. Environmental effects were minimized by installing exhaust fans producing white noise that obscured outside sounds. One-way observation windows prevented the animals from seeing outside. The sensors were turned on 45 min before drug administration (saline control or caffeine). The animals were observed for up to 8 h after i.v. administration of 2.0, 2.5, 3.0 or 5.0 mg caffeine kg(-1). The HNED of caffeine for stimulation of the spontaneous locomotor activity of horses was 2.0 mg kg(-1). The quantification of caffeine in urine and plasma samples was done by gradient HPLC with UV detection. The no-effect threshold should not be greater than 2.0 microg caffeine ml(-1) plasma or 5.0 microg caffeine ml(-1) urine.

Comparison of the quantification of caffeine in human plasma by gas chromatography and ELISA

In the present study we evaluated the precision of the ELISA method to quantify caffeine in human plasma and compared the results with those obtained by gas chromatography. A total of 58 samples were analyzed by gas chromatography using a nitrogen-phosphorus detector and routine techniques. For the ELISA test, the samples were diluted to obtain a concentration corresponding to 50 per cent of the absorbance of the standard curve. To determine whether the proximity between the I50 of the standard curve and that of the sample would bring about a more precise result, the samples were divided into three blocks according to the criterion of difference, in modulus, of the I50 of the standard curve and of the I50 of the sample. The samples were classified into three groups. The first was composed of 20 samples with I50 up to 1.5 ng/ml, the second consisted of 21 samples with I50 ranging from 1.51 to 3 ng/ml, and the third of 17 samples with I50 ranging from 3.01 to 13 ng/ml. The determination coefficient (R² = 0.999) showed that the data obtained by gas chromatography represented a reliable basis. The results obtained by ELISA were also reliable, with an estimated Pearson correlation coefficient of 0.82 between the two methods. This coefficient for the different groups (0.88, 0.79 and 0.49 for groups 1, 2 and 3, respectively) showed greater reliability for the test with dilutions closer to I50.

Comparison of the quantification of caffeine in human plasma by gas chromatography and ELISA.

In the present study we evaluated the precision of the ELISA method to quantify caffeine in human plasma and compared the results with those obtained by gas chromatography. A total of 58 samples were analyzed by gas chromatography using a nitrogen-phosphorus detector and routine techniques. For the ELISA test, the samples were diluted to obtain a concentration corresponding to 50% of the absorbance of the standard curve. To determine whether the proximity between the I50 of the standard curve and that of the sample would bring about a more precise result, the samples were divided into three blocks according to the criterion of difference, in modulus, of the I50 of the standard curve and of the I50 of the sample. The samples were classified into three groups. The first was composed of 20 samples with I50 up to 1.5 ng/ml, the second consisted of 21 samples with I50 ranging from 1.51 to 3 ng/ml, and the third of 17 samples with I50 ranging from 3.01 to 13 ng/ml. The determination coefficient (R2 = 0.999) showed that the data obtained by gas chromatography represented a reliable basis. The results obtained by ELISA were also reliable, with an estimated Pearson correlation coefficient of 0.82 between the two methods. This coefficient for the different groups (0.88, 0.79 and 0.49 for groups 1, 2 and 3, respectively) showed greater reliability for the test with dilutions closer to I50.