Effect of cooling rate and cryoprotectant on the cryosurvival of equine spermatozoa

Cryosurvival of cells is reduced if the cooling rate used is suboptimal. If cells cool too rapidly, intracellular water will freeze, causing intracellular ice crystals. However, if spermatozoa are cooled too slowly, excessive cellular dehydration occurs, causing irreversible damage to cellular compartments. In addition, cryoprotectants are added to the freezing diluent to protect cells from damage during cryopreservation. This study was conducted to determine the optimal cooling rate for stallion spermatozoa frozen in the presence of three different cryoprotectants. Spermatozoa were frozen in a skim milk, egg yolk diluent containing 4% glycerol, and ethylene glycol or dimethyl formamide at 10 different cooling rates ranging from 5°C/min to 50°C/min. The percentage of viable spermatozoa was higher for spermatozoa cooled at 10°C/min than at 50°C/min (P < .05). Spermatozoa frozen using glycerol as the cryoprotectant had higher percentages of motile and progressively motile spermatozoa compared with spermatozoa frozen using the other two cryoprotectants (P < .05). In conclusion, the cryosurvival of stallion spermatozoa is similar when cooling rates of 5°C/min to 45°C/min are used, and when 4% cryoprotectant is used, glycerol is a more effective cryoprotectant than ethylene glycol or dimethyl formamide.     

Effect of different blood‐guided conditioning programmes on skeletal muscle ultrastructure and histochemistry of sport horses

The effects of three different blood‐guided conditioning programmes on ultrastructural and histochemical features of the gluteus medius muscle of 2‐year‐old sport horses were examined. Six non‐trained Haflinger horses performed three consecutive conditioning programmes of varying lactate‐guided intensities [velocities eliciting blood lactate concentrations of 1.5 (v_1.5), 2.5 (v_2.5) and 4 (v₄) mm respectively] and durations (25 and 45 min). Each conditioning programme lasted 6 weeks and was followed by a 5‐week resting period. Pre‐, post‐ and deconditioning muscle biopsies were analysed. Although training and detraining adaptations were similar in nature, they varied significantly in magnitude among the three different conditioning programmes. Overall, the adaptations consisted in significant increases in size of mitochondria and myofibrils, as well as a hypertrophy of myofibrillar ATPase type IIA muscle fibres and a reduction in number of type IIx low‐oxidative fibres. Together, these changes are compatible with a significant improvement in both muscle aerobic capacity and muscle strength. The use of v_1.5 and v_2.5 as the exercise intensities for 45 min elicited more significant adaptations in muscle, whereas conditioning horses at v₄ for 25 min evoked minimal changes. Most of these muscular adaptations returned towards the pre‐conditioning status after 5 weeks of inactivity. It is concluded that exercises of low or moderate intensities (in the range between v_1.5 and v_2.5) and long duration (45 min) are more effective for improving muscle features associated with stamina and power in sport horses than exercises of higher intensity (equivalent to v₄) and shorter duration (25 min).     

Pharmacokinetics of xylazine after 2-, 4-, and 6-hr durations of continuous rate infusions in horses

Intravenous (i.v.) bolus administration of xylazine (XYL) (0.5 mg/kg) immediately followed by a continuous rate infusion (CRI) of 1 mg kg⁻¹ hr⁻¹ for 2, 4, and 6 hr produced immediate sedation, which lasted throughout the duration of the CRI. Heart rate decreased and blood pressure increased significantly (p > .05) in all horses during the first 15 min of infusion, both returned to and then remained at baseline during the duration of the infusion. Compartmental models were used to investigate the pharmacokinetics of XYL administration. Plasma concentration–time curves following bolus and CRI were best described by a one-compartment model. No differences were found between pharmacokinetic estimates of the CRIs for the fractional elimination rate constant (K_e), half-life (t_1/2e), volume of distribution (V_d), and clearance (Cl). Median and range were 0.42 (0.15–0.97)/hr, 1.68 (0.87–4.52) hr, 5.85 (2.10–19.34) L/kg, and 28.7 (19.6–39.5) ml min⁻¹ kg⁻¹, respectively. Significant differences were seen for area under the curve ( ) (p < .0002) and maximum concentration (C_max) (p < .04). This indicates that with increasing duration of infusion, XYL may not accumulate in a clinically relevant way and hence no adjustments are required in a longer XYL CRI to maintain a constant level of sedation and a rapid recovery.     

Doxycycline levels in preocular tear film of horses following oral administration

Objective To determine the concentration of doxycycline in preocular tear film following oral administration in horses as a possible therapeutic modality for infectious and keratomalacic equine keratitis. Procedure Eight broodmares without ocular disease from a Thoroughbred breeding facility were included in this study. Each mare received 20 mg/kg of doxycycline by mouth once daily in the morning for five consecutive days. Tears were collected 1 h after doxycycline administration starting on day one of administration and continuing for 10 consecutive days. Doxycycline levels in the tears were measured using liquid chromatography with tandem mass spectrometric detection (LC-MS/MS). Results Doxycycline was present in the tears of each mare at low µg/mL levels with the highest concentration appearing on the third to fifth days (8.21–9.83 µg/mL). Doxycycline levels had fallen below quantifiable ranges by day 10. No systemic side-effects were noted in any of the horses included in this study. Conclusions Oral doxycycline is present in preocular tear film of normal horses with noninflamed eyes and may be useful as treatment in equine ulcerative keratomalacia. The oral dose listed was tolerated well by the horses in this study. The drug levels attained at 20 mg/kg once daily orally of doxycycline may aid in the treatment of corneal ulceration in horses, but further study is warranted.     

Behavioural and cardiovascular effects of medetomidine constant rate infusion compared with detomidine for standing sedation in horses

ObjectiveTo compare the efficacy of a medetomidine constant rate infusion (CRI) with a detomidine CRI for standing sedation in horses undergoing high dose rate brachytherapy.Study designRandomized, controlled, crossover, blinded clinical trial.AnimalsA total of 50 horses with owner consent, excluding stallions.MethodsEach horse was sedated with intravenous acepromazine (0.02 mg kg^–1), followed by an α₂-adrenoceptor agonist 30 minutes later and then by butorphanol (0.1 mg kg^–1) 5 minutes later. A CRI of the same α₂-adrenoceptor agonist was started 10 minutes after butorphanol administration and maintained for the treatment duration. Treatments were given 1 week apart. Each horse was sedated with detomidine (bolus dose, 10 μg kg^–1; CRI, 6 μg kg^–1 hour^–1) or medetomidine (bolus dose, 5 μg kg^–1; CRI, 3.5 μg kg^–1 hour^–1). If sedation was inadequate, a quarter of the initial bolus of the α₂-adrenoceptor agonist was administered. Heart rate (HR) was measured via electrocardiography, and sedation and behaviour evaluated using a previously published scale. Between treatments, behaviour scores were compared using a Wilcoxon signed-rank test, frequencies of arrhythmias with chi-square tests, and HR with two-tailed paired t tests. A p value <0.05 indicated statistical significance.ResultsTotal treatment time for medetomidine was longer than that for detomidine (p = 0.04), and ear movements during medetomidine sedation were more numerous than those during detomidine sedation (p = 0.03), suggesting there may be a subtle difference in the depth of sedation. No significant differences in HR were found between treatments (p ≥ 0.09). Several horses had arrhythmias, with no difference in their frequency between the two infusions.Conclusions and clinical relevanceMedetomidine at this dose rate may produce less sedation than detomidine. Further studies are required to evaluate any clinical advantages to either drug, or whether a different CRI may be more appropriate.     

Effect of physical training on nutrient digestibility and faecal fermentative parameters in Standardbred horses

This study aimed at evaluating, in previously inactive Standardbreds horses, the effect of 5 weeks of an exercise training programme on nutrient digestibility and faecal fermentative parameters (FFPs). As an increase in digestibility had previously been reported in trained endurance horses, we hypothesized that similar results would be found in horses being trained for other types of exercise on a different type of diet. After 3 weeks of dietary adaptation, a digestibility trial (DT1) was undertaken, over 3 days, in eight untrained Standardbreds with a fresh faecal sample being collected on the second day for FFP determinations. Six of the eight horses undertook a training programme, and after 5 weeks of exercise, the DT and the FFP measurements were then repeated (DT2). DT2 began after 3 days of inactivity. The same natural meadow hay (H) and pelleted complementary feed (CF) were fed throughout. For 5 days before the DTs, horses were fed 2.1% BW on a dry matter basis (55:45 ratio H:CF). Body weight and body condition score remained constant. Apparent digestibility of dry matter, organic matter, neutral detergent fibre, hemicellulose, crude protein and gross energy, as well as faecal total volatile fatty acids (VFA), acetate and propionate concentrations were significantly (p < 0.05) higher at DT2 than at DT1. A 5‐week exercise programme had a positive impact on nutrient digestibility and FFP. Training may improve dietary energy supply, in particular via increased hindgut VFA production. The potential improvement of digestive efficiency with training should be taken into account when formulating nutritional recommendations for the exercising horse, particularly when performing light work, which is low‐intensity exercise for 1–3 h per week.     

Effect of lipopolysaccharide infusion on gene expression of inflammatory cytokines in normal horses in vivo

Horses are exquisitely sensitive to bacterial endotoxin and endotoxaemia is common in colic cases. In this study, gene expression of inflammatory cytokines was characterised in the blood of healthy horses following i.v. administration of lipopolysaccharide (LPS). Six horses received an LPS infusion and 6 controls received an equivalent volume of saline. Gene expression of genes encoding interleukin (IL)‐1α, IL‐1β, IL‐6, IL‐8, and tumour necrosis factor‐α (TNF‐α) was quantified by real‐time PCR. Gene expression of all inflammatory cytokines was upregulated following administration of LPS. Interleukin‐1α, IL‐1β, IL‐8 and TNF‐α gene expression peaked at 60 min, while IL‐6 expression peaked at 90 min post LPS infusion. Interleukin‐1β and IL‐6 messenger RNA expression levels were above the baseline values 3 h post LPS infusion, whereas IL‐1α, IL‐8 and TNF‐α expression levels returned to baseline values by 3 h after LPS infusion. It was concluded that LPS infusion upregulated gene expression of inflammatory cytokines in the blood of healthy horses.     

Effect of detomidine on visceral and somatic nociception and duodenal motility in conscious adult horses

ObjectiveTo evaluate the effects of detomidine on visceral and somatic nociception, heart and respiratory rates, sedation, and duodenal motility and to correlate these effects with serum detomidine concentrations.Study designNonrandomized, experimental trial.AnimalsFive adult horses, each with a permanent gastric cannula weighing 534 ± 46 kg.MethodsVisceral nociception was evaluated by colorectal (CRD) and duodenal distension (DD). The duodenal balloon was used to assess motility. Somatic nociception was assessed via thermal threshold (TT). Nose–to–ground (NTG) height was used as a measure of sedation. Serum was collected for pharmacokinetic analysis. Detomidine (10 or 20 μg kg^?1) was administered intravenously. Data were analyzed by means of a three–factor anova with fixed factors of treatment and time and random factor of horse. When a significant time × treatment interaction was detected, differences were compared with a simple t–test or Bonferroni t–test. Significance was set at p < 0.05.ResultsDetomidine produced a significant, dose–dependent decrease in NTG height, heart rate, and skin temperature and a significant, nondose–dependent decrease in respiratory rate. Colorectal distension threshold was significantly increased with 10 μg kg^?1 for 15 minutes and for at least 165 minutes with 20 μg kg^?1. Duodenal distension threshold was significantly increased at 15 minutes for the 20 μg kg^?1 dose. A significant change in TT was not observed at either dose. A marked, immediate decrease in amplitude of duodenal contractions followed detomidine administration at both doses for 50 minutes.Conclusions and clinical relevanceDetomidine caused a longer period of visceral anti–nociception as determined by CRD but a shorter period of anti–nociception as determined by DD than has been previously reported. The lack of somatic anti–nociception as determined by TT testing may be related to the marked decrease in skin temperature, likely caused by peripheral vasoconstriction and the low temperature cut–off of the testing device.     

The use of atropine to control heart rate responses during detomidine sedation in horses

Detomidine is a sedative-analgesic which has a pharmacological profile similar to xylazine. There is evidence that the sedative effects are mediated through alpha-2 adrenoceptors.Cardiopulmonary responses were determined using detomidine as the principal agent and as a preanesthetic prior to the induction of general anesthesia. Compatibility with guaifenesin, sodium thia-mylal and halothane were determined.As in the case of xylazine, detomidine produces a slowing of heart rates. This was found to be either sinus bradycardia or heart block. There may be a corresponding increase in systolic blood pressures. The respiratory pattern is altered through the arterial blood gases and pH data supported evidence of adequate ventilation. The heart rate response to detomidine without anticholinergic treatment was transient and related to he duration of drug action.Atropine sulfate, 0.02 mg/kg i.v. was effective in preventing or treating bradycardia or heart block from detomidine. Heart rates also increased during the administration of guaifenesin and sodium thia-mylal when given 50 min poisit-detomidine.     

Effect of age on isoproterenol-induced maximal heart rate in horses

The effect of age on maximal heart rate induced by IV infusion of isoproterenol was studied in 19 healthy, sedentary, normothermic horses ranging in age from 0.25 to 9.90 years. Isoproterenol was administered IV (1.0 micrograms/kg of body weight/min) for 3 minutes, and the heart rate attained during the last 30 seconds of the infusion was determined. Linear regression of the maximal heart rate on age suggested that the rate decreased with age in a trend described by the equation: maximal heart rate (beats/min) = 209.63 - 3.28 x age (years). The regression coefficient (r) for this relation was 0.769 (P less than 0.001). These data indicate that as healthy horses age, their beta-adrenoceptor-mediated maximal chronotropic response is diminished.