The Effect of the Acute-Phase Response on In Vitro Drug Metabolism and Plasma Protein Binding in the Horse

The effect of the acute-phase response (APR) on the activity of the hepatic drug-metabolizing system (DMS) and on the binding of phenylbutazone to plasma proteins was investigated in the horse. An APR was induced by intramuscular injections of Freund's complete adjuvant in five horses and, five days later, these horses together with five clinically normal horses were shot and the right ventral lobe of each liver removed. The hepatic microsomal fractions from the liver samples ere isolated and significantly lower (p<0.01) concentrations of cytochromes P450 and b5 and activities of aniline-p-hydroxylase and aminopyrine N-demethylase (43%, 55%, 45% and 30%, respectively) were measured in the livers from the adjuvant-inflamed horses, compared to the controls. Phenylbutazone (PBZ) was administered intravenously (4.4 mg/kg) to a further four horses and plasma protein binding was measured by ultracentrifugation. Five weeks later, these horses were injected with Freund's complete adjuvant and the intravenous administration of PBZ (4.4 mg/kg) was repeated. Inflammation induced a significant increase (p<0.01) in the unbound fraction of PBZ (5.2±0.5 as against 1.4±0.6%). These results suggest that the APR depresses the hepatic DMS and reduces the binding of PBZ to plasma proteins.     

Plasma lactate as a predictor of colonic viability and survival after 360 degrees volvulus of the ascending colon in horses

Objectives— To determine the relationship between plasma lactate concentration and colonic viability and survival in horses with ≥360° volvulus of the ascending colon.
Study Design— Retrospective study.
Animals— Horses (n=73) with ≥360° volvulus of the ascending colon.
Methods— Medical records (January 2000–November 2005) of all horses examined for colic at Michigan State University Veterinary Teaching Hospital were reviewed. Horses were included only if plasma lactate concentration was measured preoperatively and a diagnosis of ≥360° volvulus of the ascending colon was confirmed by surgery or necropsy. Non-survivors were only included if the ascending colon was evaluated histopathologically. Logistic regression analysis was used to model the relationship between lactate, colonic viability, and survival.
Results— Of 73 horses, 61 were discharged. Mean (±SD) plasma lactate concentration was significantly lower in survivors (2.98±2.53 mmol/L) compared with non-survivors (9.48±5.22 mmol/L; odds ratio [OR]=1.628, 95% confidence limit [CI]=1.259–2.105). Plasma lactate concentration was significantly lower in horses with a viable colon (3.30±2.85 mmol/L) compared with horses with a non-viable colon (9.1±6.09 mmol/L; OR=1.472, 95% CI=1.173–1.846). Plasma lactate concentration <6.0 mmol/L had a sensitivity of 84% and a specificity 83% for predicting horse survival.
Conclusions— Our results demonstrate a strong association between plasma lactate concentration at the time of hospital admission and outcome in horses with ≥360° volvulus of the ascending colon.
Clinical Relevance— Plasma lactate concentration may help predict colonic viability and horse survival after ascending colon volvulus in horses.     

Transendoscopic Chemical Ablation of Progressive Ethmoidal Hematomas in Standing Horses

Objective —To examine the response of horses with progressive ethmoidal hematoma (PEH) to intralesional injection of 4% formaldehyde solution.
Study Design —Nasal passages of horses affected with PEH were examined endoscopically at different intervals to determine the effects of intralesional injection of formaldehyde solution.
Animals —21 horses with PEH.
Methods —PEHs were injected transendoscopically with 4% formaldehyde solution. Horses were examined endoscopically and retreated at different intervals until the PEH was eliminated or was so small that reinjection was not possible.
Results —Lesions diminished significantly in size or were eliminated after 1 to 18 injections (median, 5; mean, 7.0 ± 5.62). Seventeen lesions (60.7%) resolved completely after 1 to 18 injections (median, 5; mean, 7.2 ± 5.71). Five lesions decreased markedly in size but did not resolve after receiving 1 to 18 injections (median, 5; mean, 7.6 ± 6.66). Injection of these lesions was discontinued 4.0 to 25.1 months (median, 9.5; mean, 11.02 ± 8.446) after the first injection. The PEH of one horse was removed surgically after one injection. Three horses, one with bilateral PEH, were lost to follow-up. One horse developed signs of laminitis. No other complications were observed.
Conclusions —Horses with a PEH can be treated effectively by transendoscopic, intralesional injection of 4% formaldehyde solution.     

The effect of inflammation on the disposition of phenylbutazone in Thoroughbred horses

The effect of inflammation on the disposition of phenylbutazone (PBZ) was investigated in Thoroughbred horses. An initial study ( n = 5) in which PBZ (8.8 mg/kg) was injected intravenously twice, 5 weeks apart, suggested that the administration of PBZ would not affect the plasma kinetics of a subsequent dose. Two other groups of horses were given PBZ at either 8.8 mg/kg ( n = 5) or 4.4 mg/kg ( n = 4). Soft tissue inflammation was then induced by the injection of Freud's adjuvant and the administration of PBZ was repeated at a dose level equivalent to, but five weeks later than, the initial dose. Inflammation did not appear to affect the plasma kinetics or the urinary excretion of PBZ and its metabolites, oxyphenbutazone (OPBZ) or hydroxyphenylbutazone (OHPBZ) when PBZ was administered at 8.8 mg/kg. However, small but significant increases ( P <0.05) in total body clearance ( CL _B; 29.2 ± 3.9 vs. 43.8 ± 8.1 mL/ h-kg) and the volume of distribution, calculated by area ( V _d(area); 0.18 ± 0.05 vs. 0.25 ± 0.03 L/kg) or at steady-state ( V _d(SS); 0.17±0.04 vs. 0.25 ± 0.03 L/ kg), were obtained in horses after adjuvant injection, compared to controls, when PBZ was administered at 4.4 mg/kg which corresponded to relatively higher tissues concentrations and lower plasma concentrations (calculated) at the time of maximum peripheral PBZ concentration. Soft tissue inflammation also induced a significantly ( P <0.05) higher amount of OPBZ in the urine 18 h after PBZ administration but the total urinary excretion of analytes over 48 h was unchanged. These results have possible implications regarding the administration of PBZ to the horse close to race-day.     

Humoral immune responses in the horse after intrathecal challenge with ovalbumin

Background : Diagnosis of neuro-inflammatory conditions in the horse can be challenging. Current methods include evaluation of cerebrospinal fluid (CSF) for inflammation and determination of specific antibody status. The antibody index (AI) and Goldman-Witmer coefficient (C-value) can be used to aid in the interpretation.
Hypothesis : The null hypothesis to be tested was that the AI and C-values do not change in horses with neuro-inflammation.
Animals : Twelve horses of various ages (3–17 years) and breeds (Thoroughbred, Thoroughbred cross, draft, and Arabian) were included in the study.
Methods : The study was designed as a prospective randomized study. All horses were immunized with ovalbumin in adjuvant, twice. Horses of Group 1 then were challenged by intrathecal (IT) injection of ovalbumin, whereas horses of Group 2 were challenged IM. The AI and C-values for ovalbumin and equine herpesvirus were calculated.
Results : The AI for ovalbumin increased up to 5.92 in horses after intrathecal challenge, and remained normal (<1) in horses challenged IM. The C-value for ovalbumin reached a peak of 7.48, whereas for equine herpesvirus it achieved a value of 2.69. The changes in ovalbumin C-value and AI were significantly different between days 20 and 30 in horses after intrathecal challenge at day 20 ( P = .002 and .0005, respectively).
Conclusions and Clinical Importance : The results confirm the value of the AI and C-value in the evaluation of neuro-inflammation in the horse.     

Treatment of progressive ethmoidal haematoma using intralesional injections of formalin in three horses

Three Thoroughbred horses with unilateral progressive ethmoid haematomas were treated using intralesional injections of 10% formalin (4% formaldehyde solution). Injections were performed in the standing sedated horse through the nasal passages under endoscopic guidance and, when the ethmoid haematoma involved the paranasal sinuses, through holes trephined into the affected sinus. Regression of the lesions occurred in all cases after repeated injections. This technique appears to be a safe and effective treatment for progressive ethmoid haematomas in the horse.     

Incomplete Oblique Sagittal Fractures of the Dorsal Cortex of the Third Metacarpal Bone in Six Horses

Objective —To describe incomplete oblique sagittal dorsal cortical fractures of the equine third metacarpal bone, their surgical repair, and subsequent performance of the horses.
Study Design —Retrospective examination of medical records and racing performance.
Animal Population —Six Thoroughbred race horses, 2 to 4 years of age.
Methods —Radiographic confirmation of all fractures preceded general anesthesia and surgical correction. Three fractures were treated by intracortical compression using screws placed in lag fashion, and five fractures were treated by osteostixis. Race records were reviewed for each horse to determine performance after surgery.
Results —Fractures were best observed on palmarodorsal radiographic projections. Three horses treated by intracortical compression returned to racing, but fracture recurred in one horse and was treated by osteostixis. This horse and the other three horses treated by osteostixis raced after surgery.
Conclusions —Horses with incomplete oblique sagittal fractures of the dorsal cortex of the third metacarpal bone can race after surgical management of the fracture by screws placed in lag fashion or osteostixis. The authors' preferred surgical procedure for managing this fracture is osteostixis.
Clinical Relevance —Palmarodorsal radiographic projections of the third metacarpal bone are recommended in young Thoroughbred race horses suspected of having dorsal metacarpal stress fractures.     

Effects of two large doses of equine recombinant growth hormone on clinical, haematological and serum biochemical variables in adult horses

Objective To evaluate the clinical, haematological, and serum biochemical effects of two large doses of recombinant equine growth hormone.
Design Duplicated Latin square.
Sample population
Three Thoroughbred and three Standardbred mares aged between 12 and 17 years.
Procedure Two horses were randomly assigned into one of three groups. On each of three successive days, each horse pair received one of two dosages of growth hormone or a saline placebo so that by the end of the experiment all three horse pairs had received both dosages and the saline placebo. Dose rates selected were 50 μg/kg, and 100 μg/kg. A clinical examination was performed and a venous blood sample drawn for a complete blood count and serum biochemical analysis before administration of growth hormone and at 1, 2, 3, 4, 6, 8 and 24 h after injection. Data were analysed by a repeated measures analysis of variance assessing the effects of dose and time.
Results There was an effect of time on a number of clinical, haematological, and serum biochemical variables. There were significant effects of growth hormone on heart rate and serum glucose concentration but values for both variables remained within the reference range.
Conclusion The results of the present study suggest that equine recombinant growth hormone has a wide margin of safety and show that the single administration of up to five times the recommended dose rate has no significant effects on clinical, haematological, or serum biochemical variables.     

Serum and plasma cardiac troponin I concentrations in clinically normal Thoroughbreds in training in Australia

Cardiac troponin I is a potentially useful test to identify cardiac muscle damage in the horse. Measurements of cardiac troponin I from serum or heparinised plasma samples from 23 clinically normal Thoroughbred horses in race training were analysed through a standard Australian commercial laboratory using the ADVIA Centaur Assay. The cardiac troponin I concentrations were < 0.15 microg/L from all samples. The test was then validated using macerated equine myocardium. Cardiac troponin I concentration may be useful in determining whether poor performance in Thoroughbred horses is related to active myocardial disease.     

Population distributions of phenylbutazone and oxyphenbutazone after oral and i.v. dosing in horses

Experiments to determine the residual plasma concentrations of phenylbutazone and its metabolites found in horses racing on a 'no-race day medication' or 24-h rule were carried out. One dosing schedule (oral-i.v.) consisted of 8.8 mg/kg (4 g/1000 lbs) orally for 3 days, followed by 4.4 mg/kg (2 g/1000 lbs) intravenously on day 4. A second schedule consisted of 4.4 mg/kg i.v. for 4 days. The experiments were carried out in Thoroughbred and Standardbred horses at pasture, half-bred horses at pasture, and in Thoroughbred horses in training. After administering the i.v. schedule for 4 days to Thoroughbred and Standardbred horses at pasture, the mean plasma concentrations of phenylbutazone increased from 0.77 microgram/ml on day 2 to 2.5 micrograms/ml on day 5. The shape of the frequency distribution of these populations was log-normal. These data are consistent with one horse in 1,000 yielding a plasma level of 8.07 micrograms/ml on day 5. After administration of the oral-i.v. schedule to Thoroughbred and Standardbred horses at pasture, the mean plasma concentrations of phenylbutazone were 3.4 micrograms/ml on day 2 and 3.5 micrograms/ml on day 5. The range on day 5 was from 1.4 to 8.98 micrograms/ml and the frequency distribution was log-normal. These data are consistent with one horse in 1000 having a plasma level of 15.8 micrograms/ml on day 5. In a final experiment, the oral dosing schedule was administered to 62 Thoroughbred horses in training. Plasma concentrations on day 5 in these horses averaged 5.3 micrograms/ml. The range was from 1.3 to 13.6 micrograms/ml and the frequency distribution was log-normal. Statistical projection of these values suggests that following this oral dosing schedule in racing horses about one horse in 1000 will yield a plasma level of 23.5 micrograms/ml of phenylbutazone 24 h after the last dose.     

Anesthetic Potency of Desflurane in the Horse: Determination of the Minimum Alveolar Concentration

Objective — To determine the minimum alveolar concentration (MAC) of desflurane (DES) in the horse.
Study Design — Prospective study.
Animals — Six healthy adult horses (three males and three females) weighing 370 ±16 kg and aged 9 ±2 years old.
Methods — Anesthesia was induced with DES vaporized in oxygen via a face mask connected to a large-animal, semiclosed anesthetic circle system. The horses were endotracheally intubated and positioned in right lateral recumbency. Inspired and end-tidal DES were monitored using a calibrated Ohmeda RGM 5250 multigas analyzer (Ohmeda-BOC, Spain). The MAC of desflurane that prevented gross purposeful movement in response to 60 seconds of noxious electrical stimulation of oral mucous membranes was determined.
Results — The time from the start of DES administration to lateral recumbency was 6.1 ±0.9 min. The MAC of DES in these horses was 7.6 ±0.4%. Time required for the animal to regain sternal recumbency after 98 ±4 minutes of anesthesia was 6.6 ±0.5 minutes and the time to standing was 14.3 ±2.7 minutes.
Conclusions — The MAC of desflurane in these horses was 7.6 ±0.4%. DES provided a rapid induction to, and recovery from, anesthesia.
Clinical Relevance — Desflurane offers the potential for more precise control during anesthesia, and may allow a faster and uneventful recovery. It is important to know the MAC of an inhalant to use it clinically.     

Haematological responses of repeated large volume blood collection in the horse

The haematological response of regular, repeated blood harvesting was investigated in 40 Thoroughbred and non-Thoroughbred horses that donate 8 litres of blood every 3 weeks for the purposes of commercial blood production. When this volume of blood was removed on five occasions over 12 weeks, no adverse effect on packed cell volume (PCV), haemoglobin (HB), and red blood cell count (RCC) was observed. Although PCV, RCC and Hb values decreased during the first week after blood collection, followed by a gradual increase in values until the next harvest time, all values remained within published reference ranges. Derived red cell indices [mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), and mean corpuscular haemoglobin concentration (MCHC)] also remained within reference range. We conclude that the removal of approximately 8 litres of blood (approximately 16 ml kg(-1)or 20 per cent of blood volume for a 500 kg horse) from blood donor horses every 3 weeks allows time for adequate recovery of haematological variables and does not result in adverse haematological changes.     

Pharmacokinetics of thiopentone in the horse

Abass, B.T., Weaver, B.M.Q., Staddon, G.E., Waterman, A.W. Pharmacokinetics of thiopentone in the horse. J. vet. Pharmacol. Therap . 17 , 331–338.
The pharmacokinetics of thiopentone sodium administered intravenously as a single dose (11 mg/kg) were studied in acepromazine pre-medicated horses and ponies in which anaesthesia was maintained with either halothane (Group 1) or isoflurane (Group 2). The results showed that the disposition kinetics of thiopentone in horses and ponies were best described by a three-compartment open model. In plasma, a very short initial distribution phase in both horses and ponies, half-life 1.4 ± 1.2 min (mean ± SD) and 1.3 ± 0.7 min, respectively, was obtained, which was followed by a second comparatively slower redistribution phase, half-life 16 ± 12 min and 11 ± 5 min, respectively. The volume of distribution for the drug was large, especially in the ponies which received isoflurane (1127 ± 86 ml/kg). compared to the horses which received halothane (742 ± 89 ml/kg). The drug had a somewhat shorter elimination half-life in the horses (147 ± 21 min) than in the ponies (222 ± 44 min), but no obvious difference in clearance of the drug was observed between the horses (3.5 ± 0.5 ml/min/kg) and ponies (3.6 ± 0.8 ml/min/kg).     

The pharmacokinetics of glycopyrrolate in Standardbred horses

The disposition of plasma glycopyrrolate (GLY) is characterized by a three‐compartment pharmacokinetic model after a 1‐mg bolus intravenous dose to Standardbred horses. The median (range) plasma clearance (Clp), volume of distribution of the central compartment (V₁), volume of distribution at steady‐state (Vss), and area under the plasma concentration–time curve (AUC_0‐inf) were 16.7 (13.6–21.7) mL/min/kg, 0.167 (0.103–0.215) L/kg, 3.69 (0.640–38.73) L/kg, and 2.58 (2.28–2.88) ng*h/mL, respectively. Renal clearance of GLY was characterized by a median (range) of 2.65 (1.92–3.59) mL/min/kg and represented approximately 11.3–24.7% of the total plasma clearance. As a result of these studies, we conclude that the majority of GLY is cleared through hepatic mechanisms because of the limited extent of renal clearance of GLY and absence of plasma esterase activity on GLY metabolism. Although the disposition of GLY after intravenous administration to Standardbred horses was similar to that in Thoroughbred horses, differences in some pharmacokinetic parameter estimates were evident. Such differences could be attributed to breed differences or study conditions. The research could provide valuable data to support regulatory guidelines for GLY in Standardbred horses.     

AORTO-CARDIAC FISTULAS IN SEVEN HORSES

This report describes the history, clinical, electrocardiographic and echocardiographic findings, treatment, outcome, and post-mortem findings in seven horses with aorto-cardiac fistula. Affected horses included 5 stallions, one gelding and one mare; 2 each of the Thoroughbred, Arabian and Standardbred breeds and one Thoroughbred-cross with a mean ± s.d. age of 12 ± 4 years, range 6–18 years. The presenting sings were acute distress (four horses), exercise intolerance (two horses) and the lesion was detected during a routine examination in one horse. Five horses had monomorphic ventricular tachycardia on admission and one other had a history of this arrhythmia. Five horses had a characteristic continuous murmur loudes in the right fourth intercostal space. Echocardiography (six horuses) and/or post-mortem examination (four horses) revealed the horses had aorto-cardiac fistulas arising from the right aortic sinus in all five horses in which the site was recorded. Two horses had ruptured aneurysmall dilatations of the aortic wall at this site. Fistulas extended into the right ventricle in four horses; the right atrium in two horses, the left ventricle in one horse, and five horses had dissecting tracts in the septal myocardium. Horses survived for periods ranging from 24th to 4 years. Aorto-cardiac fistula should be considered in the differential diagnosis for horses presenting with acute distress, bounding arterial pulse, a right-sided continuous murmur and/or monomorphic ventricular tachycardia, particularly in middle-aged or older stallions. Echocardiography is the technique of choice for confirming the diagnosis and demonstrating accompanying cardiac changes.     

In Vitro Model for Testing Novel Implants for Equine Laryngoplasty

Objective— To develop an in vitro laryngeal model to mimic airflow and pressures experienced by horses at maximal exercise with which to test laryngoplasty techniques.
Study Design— Randomized complete block.
Sample Population— Cadaveric equine larynges (n=10).
Methods— Equine larynges were collected at necropsy and a bilateral prosthetic laryngoplasty suture was placed with #5 Fiberwire^™ suture to achieve bilateral maximal arytenoid abduction. Each larynx was positioned in a flow chamber and subjected to static flow and dynamic flow cycling at 2 Hz. Tracheal pressure and flow, and pressure within the flow chamber were recorded at a sampling frequency of 500 Hz. Data obtained were compared with the published physiologic values for horses exercising at maximal exercise.
Results— Under static flow conditions, the testing system produced inspiratory tracheal pressures (mean±SEM) of −33.0±0.98 mm Hg at a flow of 54.48±1.8 L/s. Pressure in the flow chamber was −8.1±2.2 mm Hg producing a translaryngeal impedance of 0.56±0.15 mm Hg/L/s. Under dynamic conditions, cycling flow and pressure were reproduced at a frequency of 2 Hz, the peak inspiratory (mean±SEM) pharyngeal and tracheal pressures across all larynges were −8.85±2.5 and −35.54±1.6 mm Hg, respectively. Peak inspiratory flow was 51.65±2.3 L/s and impedance was 0.57±0.06 mm Hg/L/s.
Conclusions— The model produced inspiratory pressures similar to those in horses at maximal exercise when airflows experienced at exercise were used.
Clinical Relevance— This model will allow testing of multiple novel techniques and may facilitate development of improved techniques for prosthetic laryngoplasty.     

Emotional Response to Novelty and to Expectation of Novelty in Young Race Horses

The aim of this study was to estimate the emotional response to novelty and to expectation of novelty in young race horses. The novelty in this study was the first training on an automated horse walker at a new training center. To estimate the level of emotionality in horses, the telemetric measurement of heart rate (HR) was used. A hypothesis was developed that expectation of novelty can be as exciting for horses as a novelty test. In this study, 40 horses were studied just before and then during their first walk on an automated horse walker. They were divided into four groups, with 10 horses in each group. These groups were as follows: (1) 1.5-year-old Thoroughbred colts, (2) 1.5-year-old Thoroughbred fillies, (3) 2.5-year-old Purebred Arabian colts, and (4) 2.5-year-old Purebred Arabian fillies. HR was measured at rest before exercise, during handling and moving the horse from the stable, while walking on the automated horse walker for about 20 minutes, while moving the horse from the walker to the stable, and at rest after exercise. HR response to the anticipation of novelty was higher in colts than in fillies, particularly in the group of Thoroughbreds.     

Phenylbutazone and the horse--a review.

The clinical uses and side-effects of phenylbutazone in man, horses, and other animals are reviewed. The blood dyscrasias commonly described in man have not been reported in the horse, although several of the more minor side-effects have occasionally been seen (e.g. water retention, depression, transient staggering and phlebitis). Despite the lack of documented evidence, the toxicity of phenylbutazone in the horse is considered to be lower than that in man. This may be associated with the lower dose rates normally used, the more rapid plasma clearance rate and the comparatively younger age of most horses under treatment. The following guidelines for the use of phenylbutazone in practice are put toward. It should only be used under strict veterinary control and then only if there are clear clinical indications. It should not be given if there are signs of gastro-intestinal ulceration, clotting defects or any cardiac, renal or hepatic dysfunction. Dose rates should be kept to a minimum and the drug withdrawn immediately if any side-effects occur or if there is no clinical response within 4 days. If prolonged therapy is necessary, periodic haematological analyses should be carried out.     

Nonsteroidal anti-inflammatory agents and musculoskeletal injuries in Thoroughbred racehorses in Kentucky

Injuries sustained by horses during racing have been considered as an unavoidable part of horse racing. Many factors may be associated with the musculoskeletal injuries of Thoroughbred race horses. This study surveyed the amounts of nonsteroidal anti-inflammatory agents (NSAIDs) in injured horse's biological system (plasma) at Kentucky racetracks from January 1, 1995 through December 31, 1996. During that period, there were 84 catastrophic cases (euthanized horses) and 126 noncatastrophic cases. Plasma concentrations of NSAIDs were determined by High Performance Liquid Chromatography in injured and control horses. The possible role of anti-inflammatory agents in musculoskeletal injuries of Thoroughbred race horses was investigated by comparing the apparent concentrations of NSAIDs in injured horses to concentrations in control horses. The plasma concentrations of phenylbutazone and flunixin were higher in injured horses than in control horses. Most injured and control horses did not have a detectable level of naproxen in their plasma samples. Further studies must be carried out to determine whether horses with higher plasma concentrations of NSAIDs have an altered risk of musculoskeletal injuries compared with other horses.