Changes in Dynamic Thiol/Disulfide Homeostasis, and Substance P, B-Endorphin and α-Tocopherol Concentrations in the Spinal Cord of Chronically Lame Dairy Cows.

Initial lameness inflammation leads to chronic lameness and development of chronic pain due to the release of pro-inflammatory mediators such as reactive oxygen species (ROS), which are implicated in the transition from acute to chronic pain, and free radical scavengers countering thiol, substance P (SP), and ß-endorphin (BE). The present study aimed to evaluate the dynamic thiol-disulfide homeostasis, α-tocopherol concentrations and SP and BE concentrations in the spinal cord of chronically lame dairy cows. Ten lame and 10 non-lame cows with a parity range of 2-6 were selected for the study. Lame cows had a history of up to 3 months of lameness. Spinal cord samples were obtained from the L2 to L4 lumbar vertebrae aspect of each animal. A thiol-disulfide homeostasis assay was performed using absorbance, and the α-tocopherol concentration was determined by HPLC. SP and BE concentrations were measured using ELISA kits. The results indicated that SP and BE were significantly higher in the spinal cord of lame cows. In contrast, disulfide levels and α-tocopherol concentrations were significantly lower in the spinal cord of lame cows. In conclusion, disulfide levels and α-tocopherol concentrations indicated a defective antioxidant response in cows with chronic lameness. The results of SP and BE concentrations suggested chronic pain and a defective endogenous analgesic response.

Bovine tumor necrosis factor-alpha Increases IL-6, IL-8, and PGE2 in bovine fibroblast-like synoviocytes by metabolic reprogramming.

Lameness is a common condition in dairy cattle caused by infectious or noninfectious agents. Joint lesions are the second most common cause of lameness and can be diagnosed in association with the presentation of digit injuries. Fibroblast-like synoviocyte (FLS) are predominant cells of synovia and play a key role in the pathophysiology of joint diseases, thus increasing the expression of proinflammatory mediators. Tumor necrosis factor-alpha (TNF-α) is a potent proinflammatory cytokine involved in cyclooxygenase 2 (COX-2) and proinflammatory cytokine expression in FLS. Previously, TNF-α was demonstrated to increase hypoxia-inducible Factor 1 (HIF-1), a transcription factor that rewires cellular metabolism and increases the expression of interleukin (IL)-6 in bovine FLS (bFLS). Despite this, the proinflammatory effects of TNF-α in bFLS on metabolic reprogramming have been poorly studied. We hypothesized that TNF-α increases glycolysis and in this way controls the expression of IL-6, IL-8, and COX-2 in bFLS. Results first, gas chromatography/mass spectrometry (GC/MS)-based untargeted metabolomics revealed that bTNF-α altered the metabolism of bFLS, increasing glucose, isoleucine, leucine, methionine, valine, tyrosine, and lysine and decreasing malate, fumarate, α-ketoglutarate, stearate, palmitate, laurate, aspartate, and alanine. In addition, metabolic flux analysis using D-glucose-13C6 demonstrated an increase of pyruvate and a reduction in malate and citrate levels, suggesting a decreased flux toward the tricarboxylic acid cycle after bTNF-α stimulation. However, bTNF-α increased lactate dehydrogenase subunit A (LDHA), IL-6, IL-8, IL-1ß and COX-2 expression, which was dependent on glycolysis and the PI3K/Akt pathway. The use of FX11 and dichloroacetate (DCA), an inhibitor of LDHA and pyruvate dehydrogenase kinase (PDK) respectively, partially reduced the expression of IL-6. Our results suggest that bTNF-α induces metabolic reprogramming that favors glycolysis in bFLS and increases IL-6, IL-8, IL-1ß and COX-2/PGE2.

First Molecular Confirmation of Treponema spp. in Lesions Consistent with Digital Dermatitis in Chilean Dairy Cattle.

Digital dermatitis (DD) is a highly contagious and infectious disease in cattle which has a considerable negative economic impact worldwide, and adversely affects animal welfare. Members of the genus Treponema are the only bacterial agents for which there is consistent evidence of participation in DD lesions. In Chile, DD has been described since the 1990s, but only under a clinical approach. To date, the presence of the pathogenic agent has not been confirmed in Chile by any type of confirmatory microbiological diagnosis. The aim of the present study was to detect the presence of Treponema spp. DNA in lesions consistent with DD, in Chilean dairy cattle for the first time. We provide PCR confirmation of Treponema spp. in Chilean dairy cattle affected by DD. The high rate of positive results, as well as the proportion of the main Treponema species involved, is in line with what have been described in published studies elsewhere. Future herd control plans should benefit from the molecular detection of these pathogenic bacteria associated with DD.

Pain Management in Farm Animals: Focus on Cattle, Sheep and Pigs.

Pain causes behavioral, autonomic, and neuroendocrine changes and is a common cause of animal welfare compromise in farm animals. Current societal and ethical concerns demand better agricultural practices and improved welfare for food animals. These guidelines focus on cattle, sheep, and pigs, and present the implications of pain in terms of animal welfare and ethical perspectives, and its challenges and misconceptions. We provide an overview of pain management including assessment and treatment applied to the most common husbandry procedures, and recommendations to improve animal welfare in these species. A cost-benefit analysis of pain mitigation is discussed for food animals as well as the use of pain scoring systems for pain assessment in these species. Several recommendations are provided related to husbandry practices that could mitigate pain and improve farm animal welfare. This includes pain assessment as one of the indicators of animal welfare, the use of artificial intelligence for automated methods and research, and the need for better/appropriate legislation, regulations, and recommendations for pain relief during routine and husbandry procedures.

Intracellular potassium depletion enhances apoptosis induced by staurosporine in cultured trigeminal satellite glial cells.

PURPOSE: Satellite glial cells (SGC) surrounding neurons in sensory ganglia can buffer extracellular potassium, regulating the excitability of injured neurons and possibly influencing a shift from acute to neuropathic pain. SGC apoptosis may be a key component in this process. This work evaluated induction or enhancement of apoptosis in cultured trigeminal SGC following changes in intracellular potassium [K]ic. MATERIALS AND METHODS: We developed SGC primary cultures from rat trigeminal ganglia (TG). Purity of our cultures was confirmed using immunofluorescence and western blot analysis for the presence of the specific marker of SGC, glutamine synthetase (GS). SGC [K]ic was depleted using hypo-osmotic shock and 4 mM bumetanide plus 10 mM ouabain. [K]ic was measured using the K+ fluorescent indicator potassium benzofuran isophthalate (PBFI-AM). RESULTS: SGC tested positive for GS and hypo-osmotic shock induced a significant decrease in [K]ic at every evaluated time. Cells were then incubated for 5 h with either 2 mM staurosporine (STS) or 20 ng/ml of TNF-α and evaluated for early apoptosis and late apoptosis/necrosis by flow cytometry using annexin V and propidium iodide. A significant increase in early apoptosis, from 16 to 38%, was detected in SGC with depleted [K]ic after incubation with STS. In contrast, TNF-α did not increase early apoptosis in normal or [K]ic depleted SGC. CONCLUSION: Hypo-osmotic shock induced a decrease in intracellular potassium in cultured trigeminal SGC and this enhanced apoptosis induced by STS that is associated with the mitochondrial pathway. These results suggest that K+ dysregulation may underlie apoptosis in trigeminal SGC.

Prepartum Fat Mobilization in Dairy Cows with Equal Body Condition and Its Impact on Health, Behavior, Milk Production and Fertility during Lactation.

The objective of this study was to evaluate the effect of two levels of fat mobilization at the close-up period in dairy cows with an equal body condition score (BCS = 3.0) on the circulating concentrations of metabolic, inflammatory, and oxidative stress biomarkers, incidence of diseases, behavior, milk production, and fertility during the postpartum. Late-gestation multiparous Holstein cows (n = 59) with a body condition score of 3.0 (5-point scale) were enrolled at the beginning of the close-up period and then were followed during the entire lactation. Cows were retrospectively allocated into two groups: animals with prepartum non-esterified fatty acids concentration over 0.3 mmol/L were categorized as high fat mobilization (HFM) (n = 26), and below this threshold as low fat mobilization (LFM) (n = 33). Blood samples were collected 21 d before expected calving and once weekly for 3 wk postpartum in order to analyze ß-hydroxybutirate, haptoglobin, fibrinogen, total proteins, and malondialdehyde. Health was observed daily for 21 d postpartum. Behavioral data was collected with an accelerometer and milk production and fertility were obtained from the farm records. An increased fat mobilization in dairy cows with equal BCS modified the inflammatory and oxidative stress responses during the early postpartum without impairing their health status and fertility. Moreover, milk production and behavior were markedly affected by excessive prepartum fat mobilization through lactation.

Chronic Inflammatory Lameness Increases Cytokine Concentration in the Spinal Cord of Dairy Cows.

Lameness in dairy cows is an extremely painful multifactorial condition that affects the welfare of animals and economically impacts the dairy industry worldwide. The aim of this study was to determine the profile of cytokines in the spinal cord dorsal horn of dairy cows with painful chronic inflammatory lameness. Concentrations of 10 cytokines were measured in the spinal cord of seven adult dairy cows with chronic lameness and seven adult dairy cows with no lameness. In all cows lameness was evaluated using a mobility scoring system and registered accordingly. Immediately after euthanasia the spinal cord was removed and 20 cm of lumbar segments (L2-L5) were obtained. After dorsal horn removal and processing, cytokine quantification of tumor necrosis factor-alpha (TNF-α), interleukin-1alpha (IL-1α), interleukin 13 (IL-13), chemokine-10 (CXCL10/IP-10), chemokine-9 (CXCL9/MIG), interferon-alpha (IFN-α), interferon-gamma (IFN-γ), interleukin-21 (IL-21), interleukin-36ra (IL-36ra), and macrophage inflammatory protein-1 beta (MIP-1ß) was performed using a multiplex array. Lame cows had higher concentrations of TNF-α, IL-1-α, IL-13, CXCL10, CXCL9, IFN-α, and IFN-γ in their dorsal horn compared to non-lame cows, while IL-21 concentration was decreased. No differences in IL-36ra and MIP-1ß concentrations between lame and non-lame cows were observed. Painful chronic inflammation of the hoof in dairy cows leads to a marked increase in cytokine concentration in the dorsal horn of the spinal cord, which could represent a state of neuroinflammation of the Central Nervous System (CNS).

Proteomic profiling of proteins in the dorsal horn of the spinal cord in dairy cows with chronic lameness.

Chronic lameness affects bovine welfare and has a negative economic impact in dairy industry. Moreover, due to the translational gap between traditional pain models and new drugs development for treating chronic pain states, naturally occurring painful diseases could be a potential translational tool for chronic pain research. We therefore employed liquid chromatography tandem mass spectrometry (LC-MS/MS) to stablish the proteomic profile of the spinal cord samples from lumbar segments (L2-L4) of chronic lame dairy cows. Data were validated and quantified through software tool (Scaffold® v 4.0) using output data from two search engines (SEQUEST® and X-Tandem®). Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) analysis was performed to detect proteins interactions. LC-MS/MS identified a total amount of 177 proteins; of which 129 proteins were able to be quantified. Lame cows showed a strong upregulation of interacting proteins with chaperone and stress functions such as Hsp70 (p < 0.006), Hsc70 (p < 0.0079), Hsp90 (p < 0.015), STIP (p > 0.0018) and Grp78 (p <0.0068), and interacting proteins associated to glycolytic pathway such as; γ-enolase (p < 0.0095), α-enolase (p < 0.013) and hexokinase-1 (p < 0.028). It was not possible to establish a clear network of interaction in several upregulated proteins in lame cows. Non-interacting proteins were mainly associated to redox process and cytoskeletal organization. The most relevant down regulated protein in lame cows was myelin basic protein (MBP) (p < 0.02). Chronic inflammatory lameness in cows is associated to increased expression of stress proteins with chaperone, metabolism, redox and structural functions. A state of endoplasmic reticulum stress and unfolded protein response (UPR) might explain the changes in protein expression in lame cows; however, further studies need to be performed in order to confirm these findings.

Spinal Reactive Oxygen Species and Oxidative Damage Mediate Chronic Pain in Lame Dairy Cows.

Lameness in dairy cows is a worldwide prevalent disease with a negative impact on animal welfare and herd economy. Oxidative damage and antioxidant system dysfunction are common features of many CNS diseases, including chronic pain. The aim of this study was to evaluate the levels of reactive oxygen species (ROS) and oxidative damage markers in the spinal cord of dairy cows with chronic inflammatory lameness. Locomotion score was performed in order to select cows with chronic lameness. Dorsal horn spinal cord samples were obtained post mortem from lumbar segments (L2-L5), and ROS, malondialdehyde (MDA), and carbonyl groups were measured along with the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and total antioxidant response (TAR). Lame cows had increased levels of ROS, MDA, and carbonyl groups, while no differences were observed between lame and non-lame cows in SOD, GPx, CAT, and TAR activity. We conclude that painful chronic inflammatory lameness in dairy cows is associated with an increase in ROS, MDA, and carbonyl groups. Nonetheless, an association between ROS generation and dysfunction of the antioxidant system, as previously proposed, could not be established.

Sheep Farmers' Perception of Welfare and Pain Associated with Routine Husbandry Practices in Chile.

Considering the public concern about the welfare of farm animals during routine husbandry practices, this study aimed to determine how husbandry practices are carried out in Chilean farms, sheep farmers' perceptions of animal welfare and pain, and factors that affect them, as well as the level of agreement among farmers in the recognition of pain associated with these practices. Using a self-administered survey, participants were asked about their sociodemographic information, how husbandry practices are carried out in their farms, and their pain perception for seven of these common husbandry procedures using a numerical rating scale (0 to 10). A total of 165 farmers completed the survey and perceived castration and tail docking as the most painful practices in lambs (median pain score 10 vs. 8, p < 0.05). Pain perception was associated with the method used for the specific husbandry practices, the farmers' educational level, the farm size, and flock size (p < 0.05). There was a fair to good level of agreement beyond chance (p < 0.05) in the recognition of pain associated with the most painful practices. In general, husbandry practices are not carried out in young animals, use painful methods, without using analgesics, which may have a negative impact on animal welfare.

Plasma Concentration of Norepinephrine, ß-endorphin, and Substance P in Lame Dairy Cows.

INTRODUCTION: Lameness is a painful and debilitating condition that affects dairy cows worldwide. The aim of this study was to determine the plasma concentration of norepinephrine, ß-endorphin, and substance P in dairy cows with lameness and different mobility scores (MS). MATERIAL AND METHODS: A total of 100 Friesian and Jersey cows with lameness (parity range: 1-6; weight: 400-500 kg; milk yield: 22-28 L a day, and lactation stage less than 230 days) were selected. Animals were selected and grouped according to MS (MS 0-3; n = 25), and plasma concentration of norepinephrine, substance P, and ß-endorphin was measured using ELISA. RESULTS: Cows with MS 3 had higher plasma concentrations of norepinephrine and substance P and lower plasma concentrations of ß-endorphins when compared to MS 0 cows. CONCLUSION: Variations in plasma concentration of norepinephrine, substance P, and ß-endorphin could be associated with intense pain states in dairy cows with lameness, but are insufficient to differentiate these states from the mildest pain states. Further studies are necessary in order to evaluate the potential use of these biomarkers in the detection of chronic bovine painful conditions.

Mechanical/thermal sensitivity and superficial temperature in the stump of long-term tail-docked dairy cows.

BACKGROUND: Tail docking of dairy cows is a painful procedure that affects animal welfare level. The aims of this study were first to evaluate the response to mechanical and thermal stimulation, and second to determine the superficial temperature of the stump of tail-docked dairy cows. METHODS: One hundred and sixty-four dairy cows were enrolled. From these, 133 cows were assigned to the tail-docked (TD) group and 31 cows were selected as control animals. The following sensory assessments to evaluate pain in tail-docked cows were performed. Sensitivity of the tail region in both groups of animals was evaluated using a portable algometer. Cold and heat sensitivity assessment was performed using a frozen pack (0 °C) and warm water (45 °C), respectively. Pinprick sensitivity was evaluated using a Wartenberg neurological pinwheel. Superficial temperature was evaluated using a thermographic camera. All sensory assessments and superficial temperature were evaluated in the ventral surface of the tail stump (TD) and tail (C). RESULTS: Pressure pain threshold was lower in TD cows (5.97 ± 0.19 kg) compared to control cows (11.75 ± 0.43 kg). Heat and cold sensitivity was higher in the TD cows compared to control cows with 29% and 23% of TD cows responding positively, respectively. Similarly, after pinprick sensitivity test was performed, 93% of TD cows elicited a positive response to stimulation. Tail-docked cows had lower superficial temperature (26.4 ± 0.27 °C) compared to control cows (29.9 ± 0.62 °C). DISCUSSION: Pressure pain threshold values in both groups of animals were higher than those previously reported for TD pigs, sows and cows. In contrast, pinprick stimulation evaluates the presence of punctate mechanical hyperalgesia/allodynia, usually related to traumatic nerve injury, and this association may reveal that it is possible that these animals developed a disorder associated to the development of a tail stump neuroma and concurrent neuropathic pain, previously reported in TD lambs, pigs and dogs. Thermal sensitivity showed that TD cows responded positively to heat and cold stimulation. These findings suggest that long-term TD cows could be suffering hyperalgesia/allodynia, which may be indicative of chronic pain. Lower superficial temperature in the stump may be associated to sympathetic fiber sprouting in the distal stump, which can lead to vasoconstriction and lower surface temperatures. Further studies are needed in order to confirm neuroma development and adrenergic sprouting.

Microglia and astrocyte activation in the spinal cord of lame horses.

OBJECTIVE: To determine the microglial and astrocyte response to painful lameness in horses. STUDY DESIGN: Ionized calcium binding adaptor molecule 1 (Iba-1) and glial fibrillary acidic protein (GFAP) expression, cell density and morphology were determined through immunofluorescence within the dorsal horn of equine spinal cord. ANIMALS: A total of five adult horses with acute or chronic unilateral lameness, previously scheduled for euthanasia. METHODS: Musculoskeletal lameness was evaluated in five horses through visual evaluation according to clinical guidelines. Spinal cord samples were obtained immediately after euthanasia, and distal limb lesions were confirmed through dissection and radiography. Iba-1 immunostaining was used for detection and characterization of dorsal horn microglia. GFAP was used for immunostaining of dorsal horn astrocytes. Iba-1 and GFAP labeled cells were quantified in the dorsal horn, and intensity of fluorescence was compared between the ipsi- and contralateral dorsal horn to the affected limb, and between dorsal horn segments of all horses. RESULTS: Iba-1 expression was higher in the ipsilateral dorsal horn of the affected limb in contrast to the contralateral side dorsal horn. GFAP markers did not demonstrate increased astrocytic activity on the dorsal horn ipsilateral side to the distal limb lesion of affected horses. Horses with acute lameness predominantly had a spherical shape microglial phenotype, while cells from chronic lameness cases had variable morphology. Astrocytes evidenced small somas and large processes in both acute and chronic lameness, with higher GFAP localization in the main branches. As in the case of rodents, the localization of microglia and astrocytes in horses was mainly situated within laminae I, II and III. CONCLUSIONS AND CLINICAL RELEVANCE: Iba-1 and GFAP are functional and morphological markers of spinal microglial cells and astrocytes in horses with lameness.

Immunofluorescence characterization of spinal cord dorsal horn microglia and astrocytes in horses.

The role of glial cells in pain modulation has recently gathered attention. The objective of this study was to determine healthy spinal microglia and astrocyte morphology and disposition in equine spinal cord dorsal horns using Iba-1 and GFAP/Cx-43 immunofluorescence labeling, respectively. Five adult horses without visible wounds or gait alterations were selected. Spinal cord segments were obtained post-mortem for immunohistochemical and immunocolocalization assays. Immunodetection of spinal cord dorsal horn astrocytes was done using a polyclonal goat antibody raised against Glial Fibrillary Acidic Protein (GFAP) and a polyclonal rabbit antibody against Connexin 43 (Cx-43). For immunodetection of spinal cord dorsal horn microglia, a polyclonal rabbit antibody against a synthetic peptide corresponding to the C-terminus of ionized calcium-binding adaptor molecule 1 (Iba-1) was used. Epifluorescence and confocal images were obtained for the morphological and organizational analysis. Evaluation of shape, area, cell diameter, cell process length and thickness was performed on dorsal horn microglia and astrocyte. Morphologically, an amoeboid spherical shape with a mean cell area of 92.4 + 34 µm2 (in lamina I, II and III) was found in horse microglial cells, located primarily in laminae I, II and III. Astrocyte primary stem branches (and cellular bodies to a much lesser extent) are mainly detected using GFAP. Thus, double GFAP/Cx-43 immunolabeling was needed in order to accurately characterize the morphology, dimension and cell density of astrocytes in horses. Horse and rodent astrocytes seem to have similar dimensions and localization. Horse astrocyte cells have an average diameter of 56 + 14 µm, with a main process length of 28 + 8 µm, and thickness of 1.4 + 0.3 µm, mainly situated in laminae I, II and III. Additionally, a close association between end-point astrocyte processes and microglial cell bodies was found. These results are the first characterization of cell morphology and organizational aspects of horse spinal glia. Iba-1 and GFAP/Cx-43 can successfully immune-label microglia and astrocytes respectively in horse spinal cords, and thus reveal cell morphology and corresponding distribution within the dorsal horn laminae of healthy horses. The conventional hyper-ramified shape that is normally visible in resting microglial cells was not found in horses. Instead, horse microglial cells had an amoeboid spherical shape. Horse protoplasmic astroglia is significantly smaller and structurally less complex than human astrocytes, with fewer main GFAP processes. Instead, horse astrocytes tend to be similar to those found in rodent's model, with small somas and large cell processes. Microglia and astrocytes were found in the more superficial regions of the dorsal horn, similarly to that previously observed in humans and rodents. Further studies are needed to demonstrate the molecular mechanisms involved in the neuron-glia interaction in horses.

Stress and pain response after oligofructose induced-lameness in dairy heifers.

Lameness is one of the most painful conditions that affects dairy cattle. This study was conducted to evaluate clinical signs and plasma concentration of several pain and stress biomarkers after oligofructose-induced lameness in dairy heifers. Lameness was induced using an oligofructose overload model in 12 non-pregnant heifers. Clinical parameters and blood samples were obtained at 48 and 24 h and at 6, 12, 24, 36 and 48 h after induction of lameness. Clinical parameters included heart rate, respiratory rate, ruminal frequency and lameness score. Plasma biomarkers included cortisol, haptoglobin, norepinephrine, beta-endorphin and substance P. Differences were observed in all parameters between control and treated heifers. The plasma concentration of biomarkers increased significantly in treated animals starting 6 h after induction of lameness, reaching maximum levels at 24 h for cortisol, 48 h for haptoglobin, 6 h for norepinephrine, 12 h for substance P and at 24 h for beta-endorphin. Overall, our results confirm that lameness associated pain induced using the oligofructose model induced changes in clinical parameters and plasma biomarkers of pain and stress in dairy heifers.

Analgesia preventiva en hembras Caninas sometidas a Ovariohisterectomía: comparación del efecto analgésico de Morfina y Tramadol asociados a Xilazina / Comparison of the Preventive Analgesic Effect Between Tramadol and Morphine Associated to Xylazine in Canine Ovaryhisterectomy

El objetivo del presente estudio fue comparar y evaluar el grado de analgesia postquirúrgica otorgado por Tramadol o morfina asociados a xilazina administrados por vía intramuscular (IM) en hembras caninas sometidas a ovariohisterectomía (OVH). Se utilizaron 24 animales adultos, sin distinción de raza y edad. Previo al procedimiento quirúrgico, los animales se dividieron en forma aleatoria conformado uno de los siguientes grupos: Grupo Control: 0,5 mg/kg de xilazina, Grupo Morfina: 0,5 mg/kg de xilazina + 0,4 mg/kg de morfina y Grupo Tramadol: 0,5 mg/kg de xilazina + 3 mg/kg de Tramadol. Las cirugías se desarrollaron de forma rutinaria sin presentarse anormalidades bajo anestesia general con isoflurano. La evaluación se inició 30 minutos postextubación durante 10 horas, a intervalos de 30 minutos, las primeras 6 horas y cada 1 hora por las siguientes 4 horas. El grado de algesia fue determinado mediante una escala de valoración numérica (EVN). Las diferencias entre tratamientos se evaluaron utilizando análisis de varianza de Kruskall Wallis y el método de comparación múltiple de Wilcoxon, considerando significativo un valor de P menor a 0,05. Los valores de algesia presentaron diferencias estadísticamente significativas (P<0,05) entre las tres, tres y media, cuatro y las siete horas de estudio entre los grupos control y Tramadol. A las seis, nueve y diez horas, las diferencias estadísticamente significativas (P<0,05) se dieron entre el grupo control y el grupo morfina y entre las cuatro horas treinta minutos, las cinco horas treinta minutos y las ocho horas, estas diferencias se dieron entre el grupo control y los grupos morfina y Tramadol. Del presente estudio se puede concluir que la administración preventiva de Tramadol o morfina asociados a xilazina otorgan buen efecto analgésico y que, en base a la respuesta algésica presentada en el periodo postoperatorio, ambos fármacos son eficaces y seguros para el control del dolor agudo postquirúrgico en hembras caninas sometidas a ovariohisterectomía.

The objective of the present study was to compare and evaluate the postoperative analgesic effects after the preventive administration of Tramadol and Morphine combined with Xylazine in canine ovarihysterectomy. Twenty four adult bitches without distinction of race and age were used. Animals were randomly assigned to three different groups receiving respectively: Control Group: 0.5 mg/kg Xilazine, Tramadol Group 0.5 mg/kg xilazine + 3 mg/kg Tramadol and Morphine Group: 0.5 mg/kg Xilazine + 0.4 mg/kg Morphine. Surgeries were carried out without complications under general anaesthesia under Isoflurane. Pain evaluations started 30 minutes after removing the endotracheal tube and carried out for 10 hours every thirty minutes for the first six hours and then every hour. Pain scores were determined using a numerical rating scale. Pain scores between different treatments were analyzed using Kruskall-Wallis non-parametric analysis of variance. A value of P<0.05 was considered to be statistically significant. Pain scores showed significant differences (P<0.05) between control and Tramadol groups at third, third and a half, fourth and seventh hour. Similarly at six, nine and ten hours of evaluation significant differences (P<0.05) between control and Morphine groups were found. Finally at four hour thirty minutes, the five hours thirty minutes and the eigth hour these significant differences (P<0.05) were between control and Morphine and Tramadol groups. It can be concluded that Tramadol and Morphine are effective and safe for the control of early post surgical pain in canine ovariohysterectomy.