Safety Assessment of an Oral Therapeutic Dose of Firocoxib on Healthy Horses.

Firocoxib is a non-steroidal anti-inflammatory drug specifically formulated for veterinary medicine and selectively acts on inhibiting the cyclooxygenase 2 enzyme (COX-2). This study evaluated the possible adverse effects of administering oral therapeutic firocoxib on gastric mucosa, hematological parameters, coagulation cascade, and hepatic and renal biochemistry in healthy horses. Nine clinically healthy Arabian horses, approximately 9 years old, received 0.1 mg/kg of oral firocoxib for 14 days. The gastroscopic examination was conducted 1 day before starting treatment (D0) and two days after the last blood collection (D23). Venous blood samples were obtained for laboratory tests on day 1, immediately prior to the initiation of treatment (D1), after 7 and 14 days of treatment (D7 and D14), and 7 days after the conclusion of treatment (D21. No changes were found in the gastroscopic and hematological tests. Coagulation and serum biochemistry levels remain between these species' average values. However, the increased activated partial thromboplastin time (aPTT) and prothrombin time (PT) indicate reduced blood coagulation capacity, which contradicts the expected effect of treatment with selective COX-2 inhibitors, as these drugs theoretically promote coagulation. Administering firocoxib to horses is safe as it does not cause significant adverse reactions. Therefore, it is a suitable option for managing inflammatory conditions in these animals with attention to an unexpected adverse anti-coagulopathy effect, and further study is warranted.

Facial Expressions of Horses Using Weighted Multivariate Statistics for Assessment of Subtle Local Pain Induced by Polylactide-Based Polymers Implanted Subcutaneously.

Facial-expression-based analysis has been widely applied as a pain coding system in horses. Herein, we aimed to identify pain in horses undergoing subcutaneously polylactide-based polymer implantation. The sham group was submitted only to surgical incision. The horses were filmed before and 24 and 48 h after implantation. Five statistical methods for evaluating their facial expressions (FEs) were tested. Primarily, three levels of scores (0, 1, and 2) were applied to the seven FEs (ear movements, eyebrow tension, orbicularis tension, dilated nostrils, eye opening, muzzle tension, and masticatory muscles tension). Subsequently, the scores of the seven FEs were added (SUM). Afterwards, principal component analysis (PCoA) was performed using the scores of the seven FEs obtained using the first method. Subsequently, weights were created for each FE, based on each variable's contribution variability obtained from the PCoA (SUM.W). Lastly, we applied a general score (GFS) to the animal's face (0 = without pain; 1 = moderate pain; 2 = severe pain). The mechanical nociceptive threshold (MNT) and cutaneous temperature (CT) values were collected at the same moments. The results show no intra- or intergroup differences, when evaluating each FE separately or in the GFS. In the intragroup comparison and 48 h after implantation, the control group showed higher values for SUM, PCoA, and SUM.W, although the horses implanted with polymers displayed more obvious alterations in the CT and MNT. Our findings show that the five statistical strategies used to analyze the faces of the horses were not able to detect low-grade inflammatory pain.

Genome-wide scans for signatures of selection in Mangalarga Marchador horses using high-throughput SNP genotyping.

BACKGROUND: The detection of signatures of selection in genomic regions provides insights into the evolutionary process, enabling discoveries regarding complex phenotypic traits. In this research, we focused on identifying genomic regions affected by different selection pressures, mainly highlighting the recent positive selection, as well as understanding the candidate genes and functional pathways associated with the signatures of selection in the Mangalarga Marchador genome. Besides, we seek to direct the discussion about genes and traits of importance in this breed, especially traits related to the type and quality of gait, temperament, conformation, and locomotor system. RESULTS: Three different methods were used to search for signals of selection: Tajima's D (TD), the integrated haplotype score (iHS), and runs of homozygosity (ROH). The samples were composed of males (n = 62) and females (n = 130) that were initially chosen considering well-defined phenotypes for gait: picada (n = 86) and batida (n = 106). All horses were genotyped using a 670 k Axiom® Equine Genotyping Array​ (Axiom MNEC670). In total, 27, 104 (chosen), and 38 candidate genes were observed within the signatures of selection identified in TD, iHS, and ROH analyses, respectively. The genes are acting in essential biological processes. The enrichment analysis highlighted the following functions: anterior/posterior pattern for the set of genes (GLI3, HOXC9, HOXC6, HOXC5, HOXC4, HOXC13, HOXC11, and HOXC10); limb morphogenesis, skeletal system, proximal/distal pattern formation, JUN kinase activity (CCL19 and MAP3K6); and muscle stretch response (MAPK14). Other candidate genes were associated with energy metabolism, bronchodilator response, NADH regeneration, reproduction, keratinization, and the immunological system. CONCLUSIONS: Our findings revealed evidence of signatures of selection in the MM breed that encompass genes acting on athletic performance, limb development, and energy to muscle activity, with the particular involvement of the HOX family genes. The genome of MM is marked by recent positive selection. However, Tajima's D and iHS results point also to the presence of balancing selection in specific regions of the genome.

Effect of a lactate-guided conditioning program on heart rate variability obtained using 24-Holter electrocardiography in Beagle dogs.

The dogs' responses to training exercise are seldom monitored using physiological variables, and cardiac autonomic regulation (CAR) is a relevant determinant of endurance-training adaptation. There are studies in the literature establishing that regular exercise could interfere with CAR in dogs, measured by heart rate and vagal-derived indexes of heart-rate-variability (HRV). However, few studies were found using a prescribed training program based on the lactate threshold (LT) to determine HRV by a 24-h Holter analysis. The purpose of this study was to test whether an endurance-training program (ETP) guided individually by LT raises time-domain measures of HRV in healthy Beagle dogs. Twenty dogs were assigned to two groups: control (C) and trained (T). The dogs from group T underwent an incremental exercise test (IET) to determine their LT. Both LT and velocity corresponding to the LT (VLT) was determined by visual inspection. T group performed an eight-week endurance-training program consisting of treadmill runs set to 70-80% of the VLT. Next, dogs from the group T have submitted to IET again. The maximal velocities (Vmax) at which achieved by the trained dogs in both IETs were determined. The group S did not undergo IETs or ETP. HRV was determined by the 24-hour-Holter at rest, before and on the 2°, 4°, 6° and 8° training weeks. To examine the HR impact on HRV, standard HRV variables were normalized to prevailing HR. VLT and Vmax rose in group T, indicating an improvement of dogs' aerobic and anaerobic capacity. The normalized standard HRV indexes were relatively attenuated since these variables had a reduction in the degree of correlation concerning an average HR. The ETP resulted in decreased resting heart rate and increased time-domain indices, highlighting the log-transformed square root of the mean sum of the squared differences between R-R intervals (Ln rMSSD). The lactate-guided endurance-training program could lead to better parasympathetic cardiac modulation in Beagle dogs.

Intrabladder pressure as predictor of intra-abdominal pressure in horses.

OBJECTIVES: To investigate effects of postural changes and bladder distention on intrabladder pressure whilst estimating intra-abdominal pressure in horses. DESIGN: Two-year cohort study. Patients admitted for elective surgical procedures unrelated to gastrointestinal or genitourinary tract. SETTING: School of Agricultural and Veterinarian Sciences, Jaboticabal, SP, Brazil. ANIMALS: 20 adult horses, 11 males (stallions and geldings) and 9 females; between 3.5 and 12 years, weighing 350 to 500 kg. INTERVENTIONS: Intra-abdominal pressure was directly-recorded through abdominocentesis at the ventral midline with a fluid-filled system. Intrabladder pressure was obtained from a bladder catheter with the fluid-filled system zeroed at the level of the tuber ischia with patients in dorsal recumbency or pubic symphysis if in lateral recumbency. MEASUREMENTS AND MAIN RESULTS: Body position directly influenced intra-abdominal pressure. In dorsal recumbency, intra-abdominal pressure differed (p < 0.05) from intrabladder pressure at end-inspiration and end-expiration regardless of whether the bladder was empty or distended. There was no correlation nor association between the two pressures in this body position. In lateral recumbency a difference (p <0.05) between intra-abdominal pressure and intrabladder pressure was recorded at end-inspiration with the bladder distended with 25 ml, and at end-expiration for distension volumes of 25 ml and 50 ml. There was a strong correlation between both pressures for left and right lateral recumbency, regardless of the distension volume. Ordinary least product (OLP) regression analysis showed no fixed or proportional bias between both pressures for distension volume of 50 ml, at both end-inspiration and end-expiration. CONCLUSIONS: Indirect assessment of equine intra-abdominal pressure cannot be made in dorsal recumbency. For that purpose, patients should be in left lateral recumbency with the bladder distended with 50 ml. Values can be recorded at end-inspiration or end-expiration. RESTRICTION: Occlusion of the catheter tip by the bladder wall when minimally distended.

Lactate and glucose thresholds and heart rate deflection points for Beagles during intense exercise.

OBJECTIVE To determine whether the lactate threshold of dogs could be determined by a visual method and to assess the extent of agreement and bias among treadmill velocities for the lactate threshold as determined by visual (LTv) and polynomial (LTp) methods, glucose threshold as determined by visual (GTv) and polynomial (GTp) methods, and heart rate deflection point (HRdp) as a method for estimating the aerobic capacity of dogs. ANIMALS 18 healthy adult Beagles. PROCEDURES Each dog underwent a standardized incremental treadmill exercise test once. The test ended when the dog began to show signs of fatigue. Plasma lactate and glucose concentrations and heart rate (HR) were plotted against exercise intensity (treadmill velocity) for the duration of the test, and the LTv, GTv, and HRdp were determined visually. The LTp and GTp were determined by means of a second-order polynomial function. One-way ANOVA, Pearson correlation, Bland-Altman analyses, and ordinary least products regression were used to assess the extent of agreement and bias among the various threshold velocities. RESULTS Mean velocity did not differ significantly among the thresholds evaluated. There was a strong positive correlation between the LTv velocity and the velocity for GTv (r = 0.91), LTp (r = 0.96), GTp (r = 0.94), and HRdp (r = 0.95). CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that LTv could be determined for dogs undergoing intense exercise on a treadmill, and the treadmill velocity corresponding to the LTv was associated with the velocity for the other hallmarks of endurance. Thus, that method may be useful for prescription and evaluation of conditioning programs for dogs.

Standardization of metachromatic staining method of myofibrillar ATPase activity of myosin to skeletal striated muscle of mules and donkeys / Padronização do método de coloração metacromático da atividade da miosina ATPase miofibrilar para músculo estriado esquelético de muares e asininos

This study aims at standardizing the pre-incubation and incubation pH and temperature used in the metachromatic staining method of myofibrillar ATPase activity of myosin (mATPase) used for asses and mules. Twenty four donkeys and 10 mules, seven females and three males, were used in the study. From each animal, fragments from the Gluteus medius muscle were collected and percutaneous muscle biopsy was performed using a 6.0-mm Bergstrõm-type needle. In addition to the metachromatic staining method of mATPase, the technique of nicotinamide adenine dinucleotide tetrazolium reductase (NADH-TR) was also performed to confirm the histochemical data. The histochemical result of mATPase for acidic pre-incubation (pH=4.50) and alkaline incubation (pH=10.50), at a temperature of 37ºC, yielded the best differentiation of fibers stained with toluidine blue. Muscle fibers were identified according to the following colors: type I (oxidative, light blue), type IIA (oxidative-glycolytic, intermediate blue) and type IIX (glycolytic, dark blue). There are no reports in the literature regarding the characterization and distribution of different types of muscle fibers used by donkeys and mules when performing traction work, cargo transportation, endurance sports (horseback riding) and marching competitions. Therefore, this study is the first report on the standardization of the mATPase technique for donkeys and mules...

O presente estudo objetivou padronizar o pH e a temperatura da pré-incubação e incubação do método de coloração metacromática de myofibrillar atividade ATPase da miosina (mATPase) utilizada para asininos e muares. Vinte e quatro jumentos e 10 muares, sete machos e três fêmeas, foram usados no estudo. De cada animal, fragmentos do músculo Gluteus medius foram coletados e biópsia muscular percutânea foi realizada por meio de uma agulha Bergstrõm tipo de 6,0 mm. Em adição ao método de coloração metacromática de mATPase, a técnica de nicotinamida adenina dinucleótido tetrazólio redutase (NADH-TR) foi também realizada para confirmar os dados histoquímicos. O resultado histoquímico da mATPase por pré-incubação ácida (pH=4,50) e incubação alcalina (pH=10,50), a uma temperatura de 37ºC, foi o que proporcionou a melhor diferenciação das fibras coradas com azul de toluidina. As fibras musculares foram identificadas de acordo com as seguintes cores: tipo I (oxidativa, azul claro), tipo IIA (oxidativo-glicolítico, azul intermediário) e tipo IIX (glycolytic, azul escuro). Na literatura, não foram encontradas publicações pertinentes à caracterização e distribuição dos diferentes tipos de fibras musculares utilizadas pelos asininos e muares nos trabalhos de tração, no transporte de cargas, em provas esportivas de resistência (cavalgadas) e concursos de marcha. Dessa forma, essa pesquisa é o primeiro relato sobre padronização da técnica de mATPase para muares e asininos...

Low dose of dichloroacetate infusion reduces blood lactate after submaximal exercise in horses / Baixa dose de infusão de dicloroacetato reduz o lactato sanguíneo após exercício submáximo em cavalos

The acute administration of an indirect activator of the enzyme pyruvate dehydrogenase (PDH) in human athletes causes a reduction in blood lactate level during and after exercise. A single IV dose (2.5m.kg-1) of dichloroacetate (DCA) was administered before a submaximal incremental exercise test (IET) with five velocity steps, from 5.0 m.s-1 for 1 min to 6.0, 6.5, 7.0 and 7.5m.s-1 every 30s in four untrained mares. The blood collections were done in the period after exercise, at times 1, 3, 5, 10, 15 and 20 min. Blood lactate and glucose (mM) were determined electro-enzymatically utilizing a YSI 2300 automated analyzer. There was a 15.3% decrease in mean total blood lactate determined from the values obtained at all assessment times in both trials after the exercise. There was a decrease in blood lactate 1, 3, 5, 10, 15 and 20 min after exercise for the mares that received prior DCA treatment, with respective mean values of 6.31±0.90 vs 5.81±0.50, 6.45±1.19 vs 5.58±1.06, 6.07±1.56 vs 5.26±1.12, 4.88±1.61 vs 3.95±1.00, 3.66±1.41 vs 2.86±0.75 and 2.75±0.51 vs 2.04±0.30. There was no difference in glucose concentrations. By means of linear regression analysis, V140, V160, V180 and V200 were determined (velocity at which the rate heart is 140, 160, 180, and 200 beats/minute, respectively). The velocities related to heart rate did not differ, indicating that there was no ergogenic effect, but prior administration of a relatively low dose of DCA in mares reduced lactatemia after an IET.

A administração aguda de um ativador indireto da enzima piruvato desidrogenase (PD) em atletas da espécie humana provoca redução na concentração de lactato sanguíneo durante e após exercício. Uma dose única, intravenosa de 2.5m.kg-1 de dicloroacetato (DCA) foi administrada antes de um exercício teste incremental submáximo (ETI) com cinco etapas de velocidade sendo 5,0 ms-1 por 1 minuto e 6,0, 6,5, 7,0, e 7,5 ms-1 a cada 30 segundos em quatro éguas destreinadas. As coletas de sangue foram realizadas no período após o exercício, nos momentos de 1, 3, 5, 10, 15 e 20 min. Lactato e glicose (mM) foram determinados electro-enzimaticamente utilizando um analisador automático (YSI 2300). Houve redução de 15,3% no lactato sanguíneo total médio que foi determinado a partir dos valores obtidos em todos os momentos de avaliação em ambos os testes, após o exercício. Houve diminuição na lactatemia 1, 3, 5, 10, 15 e 20 minutos após exercício para as éguas que receberam infusão de DCA, com os respectivos valores médios de 6,31±0,90 versus 5,81±0,50, 6,45±1,19 versus 5,58±1,06, 6,07±1,56 versus 5,26±1,12, 4,88±1,61 versus 3,95±1,00, 3,66±1,41 versus 2,86±0,75 e 2,75±0,51 versus 2,04±0,30. Não houve diferença nas concentrações de glicose. Por meio de análise de regressão linear, V140, V160, V180 e V200 foram determinados (velocidades em que as taxas cardíacas alcançam 140, 160, 180 e 200 bpm, respectivamente). As velocidades relacionadas com a frequência cardíaca não diferiram, indicando que não houve efeito ergogênico, mas a administração prévia de uma dose relativamente baixa de DCA em éguas reduziu a lactatemia após um ETI.

Do hematologic constituents really increase due to endurance exercise in horses? / O exercício de resistência realmente leva ao aumento dos constituintes sanguíneos em equinos?

The aim of this study was to evaluate the behavior of blood constituents in a group of horses that successfully completed long endurance rides in tropical conditions. Jugular vein puncture was done to collect blood before, during and after rides. Data were analyzed using a mathematic approach, based on the hematocrit and blood volume where the percentual change in plasma volume was used to correct the values of each variable analyzed. Significance was inferred when P<0.05. The proposed mathematical model to assess blood constituents concentrations allowed the observation of a different pattern of the variables behavior, pointing out that the approach followed by the authors could be more sensitive than ones that did not take this routine. In conclusion, the method used in this study enabled to monitor the physiological processes that actually occur during endurance effort in tropical conditions.

Este estudo objetivou avaliar o comportamento dos componentes do sangue em cavalos que concluiram com êxito provas de enduro em condições tropicais. Punção da veia jugular foi realizada para coletar o sangue antes, durante e após o evento. Os dados foram analisados por meio de uma abordagem matemática, com base no hematócrito e volume de sangue onde a variação percentual no volume plasmático foi utilizada para corrigir os valores de cada variável analisada. O nível de significância adotado foi P<0,05. O modelo matemático proposto para avaliar as concentrações de componentes do sangue permitiu a observação de um padrão diferente no comportamento de várias variáveis plasmáticas, destacando que a abordagem seguida pelos autores é mais sensível do que aquelas que não seguiram este procedimento. Em conclusão, o método utilizado neste estudo permitiu acompanhar os processos fisiológicos que realmente ocorrem durante o esforço de resistência em condições tropicais.

Causes of death of thoroughbred racehorses at Octavio Dupont Veterinary Hospital, Brazilian Jockey Club, Rio de Janeiro / Causas de mortes de cavalos de corrida registradas no Hospital Veterinário Octávio Dupont, Jockey Clube Brasileiro, Rio de Janeiro

There are few studies that approach the epidemiology of deaths in racehorses in a broad manner. The majority focus on a specific affection or procedure. Brazil does not have a program instituted for the monitoring of deaths of horses. By means of a descriptive study in association with a multivariate analysis method, an epidemiologic profile was determined for deaths related to musculoskeletal (MS), gastrointestinal (GI), respiratory (RES) systems, neurologic origin (NEU) and sudden death (SD) for the years of 2002 to 2008, at the Octavio Dupont Veterinary Hospital-Rio de Janeiro (ODVH). Males comprised the majority of deaths and that deaths were related to, decreasing order, MS>GI>SD>NEU>RES, with respect to general mortality rate per large group of determined causes (TSPMr). The majority of deaths registered included horses aged four to five years (ID4-ID5). We observed the following correspondence relations: (3-year period = SM - ID>5 - SD; ID>5 - GI; ID4-5 - MS; SF - ID<4 - RES/ NEU); (4-year period = SM - ID>5 - GI; SF - ID<4 - NEU; ID4-5 - MS; GI - ID>5). The present study points out the importance and necessity of epidemiologic studies of lesions in horses, based on diagnosis for the recognition of predisposing factors and prevention.

Existem poucas pesquisas que abordam estudos epidemiológicos de mortes em cavalos de corrida de uma forma ampla. A maioria restringe-se a uma afecção ou procedimento específico. O Brasil não possui um programa de monitoramento de mortes instituído. Através de um estudo descritivo em associação a metodologia multivariada de análise, realizou-se um estudo epidemiológico de causa mortis relacionadas com afecções musculosqueléticas (MS), gastrointestinais (GI), sistema respiratório (RES), neurológico (NEU) e mortes súbitas (SD) durante os anos de 2002 a 2008 no Hospital Octavio Dupont, Rio de Janeiro. Os machos representaram o sexo mais afetado e a causa mortis relacionada com a taxa geral de mortalidade por grandes grupos de causas determinadas apresentou a seguinte ordem decrescentemente: MS>GI>SD>NEU>RES. A maioria dos óbitos registrados foi de cavalos na faixa etária de quatro a cinco anos (ID4-ID5). Seguidamente, observaram-se as seguintes relações de correspondência: (Triênio = SM-ID>5-SD; ID>5-GI; ID4-5-MS; SF-ID<4-RES/NEU); (Quatriênio = SM - ID>5 - GI; SF - ID<4 - NEU; ID4-5 - MS; GI - ID>5). O presente estudo aponta a importância e necessidade de estudos epidemiológicos sobre lesões em cavalos atletas, objetivando o diagnostico, reconhecimento de fatores predisponentes e prevenção.

Effects of intravenous administration of caffeine on physiologic variables in exercising horses.

OBJECTIVE: To investigate the effect of acute administration of caffeine on the athletic performance of Arabian horses. ANIMALS: 12 healthy adult Arabian horses that were trained for exercise on a treadmill. PROCEDURES: By use of a crossover study design, horses received each of the following treatments: IV administration of caffeine (5 mg/kg) and IV administration of approximately the same volume of saline (0.9% NaCl) solution. Order of treatment was randomized, and there was a 10-day interval between treatments. Thirty minutes after treatments, horses underwent an incremental exercise test (IET) on a treadmill. Blood samples were collected 15 seconds before the end of each velocity step of the IET for determination of blood lactate, blood glucose, plasma cortisol, and plasma insulin concentrations. Heart rate and hematologic variables were also analyzed. RESULTS: Velocities achieved when heart rates were 180 and 200 beats/min increased significantly in caffeine-treated horses, compared with control horses. Velocities corresponding to blood lactate concentrations of 4 and 2 mmol/L decreased significantly in caffeine-treated horses, compared with control horses. In comparison between groups, insulinemia was greater in control horses and glycemia was greater in caffeine-treated horses. Plasma cortisol concentration was significantly lowered by treatment with caffeine. CONCLUSIONS AND CLINICAL RELEVANCE: IV administration of caffeine at 5 mg/kg improved the performance of Arabian horses during intense exercise of short duration and diminished the oxidative metabolism of glucose.