Methods and Reliability of Tissue Doppler Imaging for Assessment of Left Ventricular Radial Wall Motion in Horses

Background: Noninvasive assessment of left ventricular (LV) function is incompletely studied in horses.
Objectives: The goals of this study were to investigate the feasibility, techniques, and reliability of tissue Doppler imaging (TDI) for characterization of LV radial wall motion in healthy horses.
Animals: Three Standardbreds, 3 Thoroughbreds; age 8–14 years; body weight 517–606 kg.
Methods: Repeated echocardiographic examinations were performed by 2 observers in unsedated horses using TDI. Test reliability was determined by estimating measurement variability, within-day interobserver variability, and between-day interobserver and intraobserver variability of all echocardiographic variables. Variability was expressed as coefficient of variation (CV) and the absolute value below which the difference between 2 measurements will lie with 95% probability.
Results: Assessment of LV radial wall motion by TDI was feasible in all horses. Measurement variabilities were very low (CV < 5%) to low (CV 5–15%) for most variables. Within-day interobserver variability as well as between-day interobserver and intraobserver variabilities were low to moderate (CV 16–25%) for most variables. All pulsed-wave TDI variables of systolic LV function showed very low to low variability, whereas some of the variables of LV diastolic and LA function showed moderate to high (CV > 25%) variability. Pulsed-wave TDI variables appeared more reliable than color TDI variables.
Conclusions and Clinical Importance: Measurement of TDI indices of LV function is feasible and reliable in adult Standardbred and Thoroughbred horses. The clinical relevance of LV function assessment by TDI remains to be determined.     

Methods and reliability of echocardiographic assessment of left atrial size and mechanical function in horses

OBJECTIVE: To assess the feasibility, describe the techniques, and determine the reliability of transthoracic echocardiography for characterization of left atrial (LA) size and LA mechanical function in horses. ANIMALS: 6 healthy adult horses. PROCEDURES: Repeated echocardiographic examinations were performed independently by 2 observers in standing, unsedated horses by use of 2-dimensional echocardiography, pulsed-wave flow Doppler, and tissue Doppler imaging (TDI) techniques. Test reliability was determined by estimating measurement variability, within-day interobserver variability, and between-day inter- and intraobserver variability of all echocardiographic variables. Variability was expressed as the coefficient of variation (CV) and the absolute value below which the difference between 2 measurements will lie with 95% probability. RESULTS: Most echocardiographic variables of LA size had low overall variability (CV, < 15%). Among the 2-dimensional indices of LA mechanical function, area-based and volume-based ejection phase indices had moderate between-day variability (CV usually < 25%). Transmitral Doppler flow indices were characterized by low to high between-day variability (CV, 6% to 35%). The TDI wall motion velocities had high between-day variability (CV, > 25%), whereas most TDI-derived time intervals had low variability (CV, < 15%). CONCLUSIONS AND CLINICAL RELEVANCE: LA size and mechanical function can be reliably assessed in standing, unsedated horses by use of 2-dimensional echocardiography, transmitral blood flow velocity profiles, and analyses of LA wall motion by use of TDI. These results may provide useful recommendations for echocardiographic assessment of LA size and function in horses.     

Determination of tear break-up time reference values and ocular tolerance of tetracaine hydrochloride eyedops in healthy horses

Reasons for performing study: Tetracaine hydrochloride (THCl) has been reported to cause irritation in dogs. In man, some topical anaesthetics have been shown to disrupt the tear film. Tear break‐up time (TBUT) is a useful test allowing an assessment of the quality of the precorneal tear film. Only one TBUT value has been reported in horses with no information on the technique used. Objectives: To provide a method for performing the TBUT in horses and to report any side effects of a single application of THCl in clinically normal horses, particularly on the stability of the tear film. Methods: In Study 1, one drop of 0.5 or 1% THCl was applied to one eye of 20 horses divided in 2 groups. Treated eyes were assessed for the development of side effects 2.5 and 5 min after treatment. In Study 2, the TBUT was measured in both eyes of 2 groups of 10 horses, before and 2.5 and 5 min after, instillation of one drop of either 0.5 or 1% THCl. Results: No animals developed any ocular side effect after instillation. Basal TBUT was 8.3 ± 1.3 s. TBUT decreased from baseline 5 and 2.5 min after application of one drop of 0.5% THCl and one drop of 1% THCl, respectively. Conclusions: A technique to measure the TBUT in healthy horses is described and normal range values that could be used as a reference were obtained. Potential relevance: THCl is well tolerated in horses but lowers the TBUT.     

Surgery: Short‐ and long‐term evaluation of surgical treatment of strangulating obstructions of the small intestine in horses: A review of 224 cases

Summary

A retrospective study was carried out of 224 horses operated for strangulating small intestine obstructions. Fifty‐four horses were euthanized and 5 horses died during surgery which means that 165 (73%) were allowed to recover. Of these, 53 horses were euthanized or died in the clinic and 112 (50%) were discharged from the hospital. Of 90 horses available for follow‐up 1 year postoperatively, 76 (84%) were still alive. The most important causes of death or reasons for euthanasia in the direct post‐operative period were post‐operative paralytic ileus, (adhesive) peritonitis and intra‐abdominal haemorrhage. After discharge from the hospital the reasons were (adhesive) peritonitis and (recurrent) colic. Of the horses which survived for at least 1 year, 16% sometimes suffered from colic, 12% experienced problems with incisional woundhealing and 4% suffered from jugular vein thrombosis. All were in good or reasonable condition and 88% performed at (approximately) the same level as before the operation.

The type of surgical intervention (i.e. enterotomy, enterectomy) did not significantly influence the outcome of surgery, whereas the type of anastomosis did. End‐to‐end jejunojejunostomy had a better prognosis than side‐to‐side jejunocaecostomy.

It was concluded that strangulating obstructions of the small intestine still carry a poor to guarded prognosis. Mortality was highest in the direct peri‐operative period. Once discharged from hospital, prognosis can be considered to be fair to good. Attempts to improve outcome should be directed at a better handling of the ileal stump during surgery and at the prevention of post‐operative ileus and the formation of adhesions.     

Serologic prevalence of Toxoplasma gondii in horses slaughtered for food in North America.

Serum samples from 1788 horses slaughtered for food in North America were tested for antibodies to Toxoplasma gondii using the modified direct agglutination test (MAT). Antibodies to T. gondii were found by the MAT in 124 (6.9%) of 1788 sera; the titers were 1:20 (69 horses), 1:40 (37 horses), 1:80 (9 horses), and > or =1:160 (9 horses). A total of 339 selected horses were also tested by the Sabin-Feldman dye test (DT). Dye test antibodies were found in 54 horses with titers of 1:10 (29 horses) 1:20 (12 horses), 1:40 (4 horses) and 1:80 (9 horses). There was no correlation between the DT and the MAT.     

Prevalence of gastric ulcer syndrome in high-level endurance horses

Reasons for performing the study: Equine gastric ulcers syndrome (EGUS) prevalence studies are rare in the endurance horse and none has been carried out to date in horses competing at high level. Objectives: To determine the prevalence of EGUS in high‐level endurance horses. Methods: Thirty endurance horses competing at high level were selected and submitted to 2 gastroscopic examinations. The first gastroscopy was performed during the interseason period, and the second during the competition season within 2–3 days following a ride of 90–160 km. Data related to housing, feeding, training system as well as age, breed and gender were recorded for each horse. Results: The prevalence of squamous gastric ulcers was 48% during the interseason period (mean score 0.85 ± 0.13 on a scale from 0–4) and 93% during the competition season (mean score 1.60 ± 1.15) with a highly significant difference between the seasons (P = 0.001). Most of the lesions were situated in the squamous portion of the stomach but 33.3% of horses showed also glandular lesions. Significant influence of training and performance level on the gastric score (according to the distance of the ride preceding the second gastroscopy) was shown (P = 0.038). There was also a significant influence of housing on the gastric score (P = 0.002) showing higher scores in the horses kept totally on pasture (mean score: 2.14 ± 0.14) vs. horses housed in a mixed environment (stable + pasture, mean score: 1.43 ± 0.17). Both the pastured and (stabled + pastured) groups were receiving a grain supplement but the pastured group received a higher daily starch intake, which might explain the higher ulcer prevalence. There was no influence of age, breed or gender on the gastric scores. Conclusions: The high prevalence of gastric lesions observed in this study is probably related to the high level of training and performance of the horses examined compared to the population of horses included in the prior study. Potential relevance: This prevalence of EGUS in high‐level endurance horses is comparable to the prevalence established in racing horses. The equine veterinarians may take into consideration EGUS as a potential cause of poor performance.     

The beta-agonist clenbuterol in mane and tail hair of horses

REASONS FOR PERFORMING STUDY: The beta2-agonist clenbuterol is commonly administered for therapeutic purposes in the horse, but its use an an anabolic agent is illegal. Clenbuterol can be detected in blood and urine for a relatively short period after administration and detection in hair could enhance the analytical range and be used to determine the history of clenbuterol application. HYPOTHESIS: That detection in mane or tail hair is possible over an extended period. METHODS: Four horses received 0.8 microg clenbuterol hydrochloride/kg bwt b.i.d. for 10 days. Four other horses were used as untreated controls. Blood, urine, mane and tail hair samples were taken on Day 0 (before) and 5, 10, 30, 35, 40, 60, 90, 120, 150 and 360 days after start of treatment. Gas chromotography/high resolution mass spectrometry (GC/HRMS) was developed for clenbuterol analysis: limit of detection was 0.2 pg/mg; intra-assay repeatability limit r = 0.06 (confidence level 95%); interassay repeatability limit r = 0.03 (confidence level 95%). Prior to treatment, clenbuterol was absent from all samples analysed. RESULTS: Clenbuterol was detectable as early as Day 5 in tail and mane hair of Segment 1 (0-20 mm from the roots) and was maximal on Day 90. However, as time progressed, shift into lower 20 mm segments was observed. On Day 360, the maximum concentration (up to 21 pg/mg) was located in Segment 13, i.e. 26-28 cm from roots of hair. Clenbuterol was not detectable in blood or urine after Day 30. Mane and tail hair results were very similar. CONCLUSIONS: The study showed that the beta-agonist clenbuterol can be found in mane and tail hair of horses after extended periods. POTENTIAL RELEVANCE: It will be possible to detect clenbuterol in breeding and show horses where anabolic drugs have been used illegally to improve conformation. This method may also be helpful to monitor therapeutic clenbuterol treatment.     

Effect of intravenous lidocaine administration on laminar inflammation in the black walnut extract model of laminitis

Reasons for performing study: Laminitis is a serious complication of horses suffering from sepsis/endotoxaemia‐related events. Laminitis in horses and organ injury in human sepsis are both reported to involve inflammatory injury to the laminae/organs including early activation of endothelium and leucocytes leading to emigration of neutrophils into the tissue interstitium. In the black walnut extract (BWE) model, systemic inflammatory events coincide with marked increase in laminar mRNA concentrations of inflammatory genes including proinflammatory cytokines (i.e. IL‐1β, IL‐6), COX‐2, chemokines (i.e. IL‐8) and endothelial adhesion molecules (i.e. ICAM‐1 and E‐selectin). In models of human sepsis, i.v. lidocaine has been reported to decrease leucocyte and endothelial activation, and the expression of proinflammatory cytokines and chemokines. Objectives: To evaluate the effect of i.v. lidocaine therapy on the inflammatory processes documented to occur in the BWE model of laminitis. Methods: Twelve horses were administered BWE and treated immediately with either lidocaine (1.3 mg/kg bwt bolus, followed by 0.05 mg/kg bwt/min CRI, n = 6) or saline (n = 6) for 10 h. At 10 h post BWE administration, laminar samples were obtained under general anaesthesia for assessment of proinflammatory gene expression (using RT‐qPCR) and leucocyte emigration (via CD13 immunohistochemistry). At 0, 3 and 10 h post BWE administration, skin samples were obtained for assessment of leucocyte emigration (via calprotectin immunohistochemistry). Results: No significant differences between groups were noted for inflammatory gene mRNA concentrations (IL‐1β, IL‐6, IL‐8, COX‐2) or for number of leucocytes present within the laminar interstitium or skin dermis. Increased (P<0.05) laminar E‐selectin mRNA concentrations were present in the LD group (vs. SAL group). Conclusions: Continuous administration of i.v. lidocaine does not inhibit inflammatory events in either the laminae or skin in the horse administered black walnut extract. Potential relevance: This work questions the use of continuous i.v. administration of lidocaine as an effective anti‐inflammatory therapy for systemic inflammation.     

Bacterial culture of septic synovial structures of horses: Does a positive bacterial culture influence prognosis?

Reasons for performing study: The influence of synovial fluid culture on short‐ and long‐term prognosis of cases with septic synovitis requires study. Hypotheses: Horses with a positive bacterial culture from septic synovial fluid are less likely to survive or return to successful athletic function than those with a negative bacterial culture from septic synovial fluid. Methods: Records of mature horses presented to 2 equine referral hospitals for investigation of suspected septic synovitis were examined. Horses (n = 206) were included in the study if synovial fluid was submitted for full laboratory examination, including bacterial culture. A diagnosis of septic synovitis was based on a nucleated cell count >30 × 10⁹ cells/l or >90% neutrophils and other clinical, cytological and bacteriological parameters. Long‐term follow‐up was obtained by telephone questionnaire. Univariate analysis, using the Fisher's exact test, was used for all outcomes. Results: Fourteen (20.9%) of 67 horses with a positive bacterial culture from synovial fluid were subjected to euthanasia because of persistent synovial sepsis compared to 2 (1.44%) of 139 with negative bacterial cultures (P<0.001). Overall survival and successful long‐term return to function in horses with a positive bacterial culture was 50% (24/48 horses) compared to 70.5% (74/105) in culture negative horses (P = 0.01). In horses that survived to be discharged, successful long‐term return to function was not significantly different between culture positive and culture negative groups. Growth of Staphylococcus aureus from synovial fluid did not affect short‐term survival to discharge from the hospital compared to other positive bacterial culture; however, successful long‐term return to function was only 30.4% (4/13) in horses from which S. aureus was cultured compared to 73.9% (17/23) of horses in which other bacteria were cultured (P = 0.015). Conclusions and potential clinical relevance: Horses with a positive bacterial culture from a septic synovitis have a poorer prognosis for survival to discharge from hospital and overall long‐term return to function than horses that yielded no bacterial growth. When S. aureus was cultured, the long‐term prognosis was poorer.     

Pharmacokinetics of meloxicam in plasma and urine of horses

OBJECTIVE: To determine pharmacokinetic parameters for meloxicam, a nonsteroidal anti-inflammatory drug, in horses. ANIMALS: 8 healthy horses. PROCEDURE: In the first phase of the study, horses were administered meloxicam once in accordance with a 2 x 2 crossover design (IV or PO drug administration; horses fed or not fed). The second phase used a multiple-dose regimen (daily oral administration of meloxicam for 14 days), with meloxicam administered at the recommended dosage (0.6 mg/kg). Plasma and urine concentrations of meloxicam were measured by use of validated methods with a limit of quantification of 10 ng/mL for plasma and 20 ng/mL for urine. RESULTS: Plasma clearance was low (mean +/- SD; 34 +/- 0.5 mL/kg/h), steady-state volume of distribution was limited (0.12 +/- 0.018 L/kg), and terminal half-life was 8.54 +/- 3.02 hours. After oral administration, bioavailability was nearly total regardless of feeding status (98 +/- 12% in fed horses and 85 +/- 19% in nonfed horses). During once-daily administration for 14 days, we did not detect drug accumulation in the plasma. Meloxicam was eliminated via the urine with a urine-to-plasma concentration that ranged from 13 to 18. Concentrations were detected for a relatively short period (3 days) after administration of the final daily dose. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study support once-daily administration of meloxicam regardless of the feeding status of a horse and suggest a period of at least 3 days before urine concentrations of meloxicam reach concentrations that could be used in drug control programs.     

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.     

Real‐time RT‐PCR for the detection and quantitative analysis of equine rhinitis viruses

Reasons for performing study: Equine rhinitis viruses (ERV) cause respiratory disease and loss of performance in horses. It has been suggested that the economic significance of these viruses may have been underestimated due to insensitive methods of detection. Objectives: To develop a sensitive, rapid, real‐time RT‐PCR (rRT‐PCR) assay suitable for the routine diagnosis and epidemiological surveillance of the A and B variants of ERV. Methods: TaqMan primer probe sets for ERAV and ERBV were designed from conserved regions of the 5′ UTR of the ERV genome. Over 400 samples from both clinically affected and asymptomatic horses were employed for validation of the assays. ERAV samples positive by rRT‐PCR were verified by virus isolation and ERBV positive samples were verified by rRT‐PCR using a different set of primers. Results: The detection limit of the rRT‐PCR for both viruses was 10–100 genome copies. Of 250 archival nasal swabs submitted for diagnostic testing over a 7 year period, 29 were ERAV positive and 3 were ERBV positive with an average incidence rate per year of 10 and 1.5%, respectively. There was evidence of co‐circulation of ERAV and ERBV with equine influenza virus (EIV). Of 100 post race urine samples tested, 29 were ERAV positive by rRT‐PCR. Partial sequencing of 2 ERBV positive samples demonstrated that one was 100% identical to ERBV1 from a 270 bp sequence and the other was more closely related to ERBV2 than ERBV1 (95% compared to 90% nucleotide identity in 178 bp). Conclusions: The rRT‐PCR assays described here are specific and more sensitive than virus isolation. They have good reproducibility and are suitable for the routine diagnosis of ERAV and ERBV. Potential relevance: These assays should be useful for investigating the temporal association between clinical signs and rhinitis virus shedding.     

Disposition, elimination, and bioavailability of phenytoin and its major metabolite in horses

OBJECTIVE: To determine pharmacokinetics and excretion of phenytoin in horses. ANIMALS: 6 adult horses. PROCEDURE: Using a crossover design, phenytoin was administered (8.8 mg/kg of body weight, IV and PO) to 6 horses to determine bioavailability (F). Phenytoin also was administered orally twice daily for 5 days to those same 6 horses to determine steady-state concentrations and excretion patterns. Blood and urine samples were collected for analysis. RESULTS: Mean (+/- SD) elimination half-life following a single IV or PO administration was 12.6+/-2.8 and 13.9+/-6.3 hours, respectively, and was 11.2+/-4.0 hours following twice-daily administration for 5 days. Values for F ranged from 14.5 to 84.7%. Mean peak plasma concentration (Cmax) following single oral administration was 1.8+/-0.68 microg/ml. Steady-state plasma concentrations following twice-daily administration for 5 days was 4.0+/-1.8 microg/ml. Of the 12.0+/-5.4% of the drug excreted during the 36-hour collection period, 0.78+/-0.39% was the parent drug phenytoin, and 11.2+/-5.3% was 5-(phydroxyphenyl)-5-phenylhydantoin (p-HPPH). Following twice-daily administration for 5 days, phenytoin was quantified in plasma and urine for up to 72 and 96 hours, respectively, and p-HPPH was quantified in urine for up to 144 hours after administration. This excretion pattern was not consistent in all horses. CONCLUSIONS AND CLINICAL RELEVANCE: Variability in F, terminal elimination-phase half-life, and Cmax following single or multiple oral administration of phenytoin was considerable. This variability makes it difficult to predict plasma concentrations in horses after phenytoin administration.     

Intratracheal clenbuterol in the horse: its pharmacological efficacy and analytical detection

Clenbuterol, a beta2 agonist/antagonist, is the only bronchodilator approved by the US Food and Drug Administration for use in horses. The Association of Racing Commissioners International classifies clenbuterol as a class 3 agent, and, as such, its identification in post-race samples may lead to sanctions. Anecdotal reports suggest that clenbuterol may have been administered by intratracheal (IT) injection to obtain beneficial effects and avoid post-race detection. The objectives of this study were (1) to measure the pharmacological efficacy of IT dose of clenbuterol and (2) to determine the analytical findings in urine in the presence and absence of furosemide. When administered intratracheally (90 microg/horse) to horses suffering from chronic obstructive pulmonary disease (COPD), clenbuterol had effects that were not significantly different from those of saline. In parallel experiments using a behavior chamber, no significant effects of IT clenbuterol on heart rate or spontaneous locomotor activity were observed. Clenbuterol concentrations in the urine were also measured after IT dose in the presence and absence of furosemide. Four horses were administered i.v. furosemide (5 mg/kg), and four horses were administered saline (5 mL). Two hours later, all horses were administrated clenbuterol (IT, 90 microg), and the furosemide-treated horses received a second dose of furosemide (2.5 mg/kg, i.v.). Three hours after clenbuterol dose (1 h after hypothetical 'post-time'), the mean specific gravity of urine samples from furosemide-treated horses was 1.024, well above the 1.010 concentration at which furosemide is considered to interfere with drug detection. There was no interference by furosemide with 'enhanced' ELISA screening of clenbuterol equivalents in extracted and concentrated samples. Similarly, furosemide had no effect on mass spectral identification or quantification of clenbuterol in these samples. These results suggest that the IT dose of clenbuterol (90 microg) is, in pharmacological terms, indistinguishable from the dose of saline, and that, using extracted samples, clenbuterol dose is readily detectable at 3 h after dosing. Furthermore, concomitant dose of furosemide does not interfere with detection or confirmation of clenbuterol.     

Elimination of doxepin isomers from the horse following intravenous application

The tricyclic antidepressant doxepin, representing a 5:1 mixture of trans- and cis-isomers, owns tranquilizing properties. This compound has been associated with illicit medication of racing horses, and therefore should be considered in doping control. Because analysis of doxepin in equine body fluids has not been documented in the literature, a highly sensitive analytical method was developed to individually monitor the doxepin isomers in blood and urine of horses by the use of gas chromatography/mass spectrometry. Following a dose of 1 mg doxepin-HCl/kg intravenously (i.v.), both the isomers were quantified for up to 24 h in serum of horses (n=4). The beta-half-lives of the trans- and cis-isomers were 3.5 and 3.1 h, respectively. The ratio of the trans/cis-isomers was found to be constant (4.7:1) during drug elimination and thus corresponded to the original composition of the antidepressant. Up to 12 h following administration low trans-isomer concentrations in an average range of 2-6 ng/mL were detected in urine of each of the horses, while the cis-isomer was only present in two of four horses for up to 8 and 12 h, respectively. In serum, mean trans-isomer concentrations exceeded urine levels maximally 120-fold after 3 h and at least sixfold after 12 h. As serum exhibits considerably higher concentrations of the doxepin isomers as compared with urine, blood of horses is the recommended body fluid when screening for the antidepressant.     

Plasma concentrations,behavioural and physiological effects following intravenous and intramuscular detomidine in horses

Reasons for performing study: Detomidine hydrochloride is used to provide sedation, muscle relaxation and analgesia in horses, but a lack of information pertaining to plasma concentration has limited the ability to correlate drug concentration with effect. Objectives: To build on previous information and assess detomidine for i.v. and i.m. use in horses by simultaneously assessing plasma drug concentrations, physiological parameters and behavioural characteristics. Hypothesis: Systemic effects would be seen following i.m. and i.v. detomidine administration and these effects would be positively correlated with plasma drug concentrations. Methods: Behavioural (e.g. head position) and physiological (e.g. heart rate) responses were recorded at fixed time points from 4 min to 24 h after i.m. or i.v. detomidine (30 μg/kg bwt) administration to 8 horses. Route of administration was assigned using a balanced crossover design. Blood was sampled at predetermined time points from 0.5 min to 48 h post administration for subsequent detomidine concentration measurements using liquid chromatography‐mass spectrometry. Data were summarised as mean ± s.d. for subsequent analysis of variance for repeated measures. Results: Plasma detomidine concentration peaked earlier (1.5 min vs. 1.5 h) and was significantly higher (105.4 ± 71.6 ng/ml vs. 6.9 ± 1.4 ng/ml) after i.v. vs. i.m. administration. Physiological and behavioural changes were of a greater magnitude and observed at earlier time points for i.v. vs. i.m. groups. For example, head position decreased from an average of 116 cm in both groups to a low value 35 ± 23 cm from the ground 10 min following i.v. detomidine and to 64 ± 24 cm 60 min after i.m. detomidine. Changes in heart rate followed a similar pattern; low value of 17 beats/min 10 min after i.v. administration and 29 beats/min 30 min after i.m. administration. Conclusions: Plasma drug concentration and measured effects were correlated positively and varied with route of administration following a single dose of detomidine. Potential relevance: Results support a significant influence of route of administration on desirable and undesirable drug effects that influence case management.     

Thromboelastography in healthy horses and horses with inflammatory gastrointestinal disorders and suspected coagulopathies

Objectives – To evaluate the use of citrated recalcified (nonactivated) thromboelastography (TEG) in healthy horses and horses with colitis and suspected coagulopathies. Design – Prospective, observational study conducted between October 2007 and June 2009. Setting – Veterinary Teaching Hospital. Animals – Forty‐five healthy adult horses and 12 sick adult horses with colitis and prolonged prothrombin time (PT) or activated partial thromboplastin time (aPTT). Interventions – None. Measurements and Main Results – Whole blood was collected on admission. Coagulation profile (PT, aPTT, platelet count, and fibrinogen concentration) and citrated recalcified whole blood TEG analysis (R‐time [R], K‐time [K], angle [α], maximum amplitude [MA], G value [G], lysis at 60 min [LY60]) were evaluated. Mean values (SD) for TEG parameters in healthy horses were: R=10.4 (3.1) minutes; K=3.5 (1.2) minutes; α=46.3 (11.0)°; MA=55.6 (5.1) mm; G=6,429 (1,341) dyn/cm², and LY60=5.1 (2.4)%. Mean coefficients of variation for intra‐assay/interindividual variability in healthy horses were: R=4.7%/30.7%, K=4.8%/35.3%, α=4.4%/23.8%, MA=1.4%/9.3%, G=3.4%/20.8%, and LY60=13.1%/47.7%, respectively. Horses with colitis and prolonged PT and/or aPTT had longer mean values for R (P<0.001) and K (P<0.001), narrower mean α (P<0.001), decreased mean MA (P=0.001), and smaller mean G (P=0.02); changes consistent with hypocoagulability. Conclusions – Citrated recalcified (nonactivated) TEG demonstrated changes consistent with hypocoagulability in horses with colitis that had preidentified coagulation abnormalities. This technique has high interindividual variability and low intra‐assay variability. TEG may be useful for detecting hypocoagulable states in horses with colitis and suspected coagulopathies.     

Pharmacokinetics and tissue fluid distribution of cephalexin in the horse after oral and i.v. administration

The purpose of this study was to determine the pharmacokinetics and tissue fluid distribution of cephalexin in the adult horse following oral and i.v. administration. Cephalexin hydrate (10 mg/kg) was administered to horses i.v. and plasma samples were collected. Following a washout period, cephalexin (30 mg/kg) was administered intragastrically. Plasma, interstitial fluid (ISF) aqueous humor, and urine samples were collected. All samples were analyzed by high-pressure liquid chromatography (HPLC). Following i.v. administration, cephalexin had a plasma half-life (t(1/2)) of 2.02 h and volume of distribution [V(d(ss))] of 0.25 L/kg. Following oral administration, the average maximum plasma concentration (C(max)) was 3.47 mug/mL and an apparent half-life (t(1/2)) of 1.64 h. Bioavailability was approximately 5.0%. The AUC(ISF):AUC(plasma) ratio was 80.55% which corresponded to the percentage protein-unbound drug in the plasma (77.07%). The t(1/2) in the ISF was 2.49 h. Cephalexin was not detected in the aqueous humor. The octanol:water partition coefficient was 0.076 +/- 0.025. Cephalexin was concentrated in the urine with an average concentration of 47.59 microg/mL. No adverse events were noted during this study. This study showed that cephalexin at a dose of 30 mg/kg administered orally at 8 h dosage intervals in horses can produce plasma and interstitial fluid drug concentrations that are in a range recommended to treat susceptible gram-positive bacteria (MIC < or = 0.5 microg/mL). Because of the low oral bioavailability of cephalexin in the horse, the effect of chronic dosing on the normal intestinal bacterial flora requires further investigation.     

A 90‐day adaptation to a high glycaemic diet alters postprandial lipid metabolism in non‐obese horses without affecting peripheral insulin sensitivity

High glycaemic feeds are associated with the development of insulin resistance in horses. However, studies that evaluated the effect of high glycaemic feeds used horses that either ranged in body condition from lean to obese or were fed to increase body condition over a period of months; thus, the ability of high glycaemic feeds to induce insulin resistance in lean horses has not been determined. This study evaluated the insulin sensitivity of 18 lean horses fed a 10% (LO; n = 6), 20% (MED; n = 6) or 60% (HI; n = 6) non‐structural carbohydrate complementary feed for 90 days. Although both the MED and HI diets increased insulinaemic responses to concentrate feeding in relation to the LO diet (p > 0.05), neither induced insulin resistance, as assessed by glucose tolerance test, following the 90‐day feeding trial. Interestingly, the post‐feeding suppression of plasma non‐esterified fatty acids was less pronounced in HI‐fed horses (p = 0.054) on days 30 and 90 of the study, potentially indicating that insulin‐induced suppression of adipose tissue lipolysis was reduced. As insulin‐resistant animals often have elevated plasma lipid concentrations, it is possible that altered lipid metabolism is an early event in the development of insulin resistance. The effects of high glycaemic feeds that are fed for a longer duration of time, on glucose and lipid metabolism, should be investigated further.     

Predictive value of hypoglycin A and methylencyclopropylacetic acid conjugates in a horse with atypical myopathy in comparison to its cograzing partners

Hypoglycin A (HGA) was detected in blood and urine of a horse suffering from atypical myopathy (AM; Day 2, serum, 8290 μg/l; urine: Day 1, 574, Day 2, 742 μg/l) and in its cograzing partners with a high variability (46–1570 μg/l serum). Over the period of disease, the level of the toxic metabolites (methylencyclopropylacetic acid [MCPA]‐conjugates) increased in body fluids of the AM horse (MCPA‐carnitine: Day 2, 0.246, Day 3, 0.581 μmol/l serum; MCPA‐carnitine: Day 2, 0.621, Day 3, 0.884 μmol/mmol creatinine in urine) and HGA decreased rapidly (Day 3, 2430 μg/l serum). In cograzing horses MCPA‐conjugates were not detected. HGA in seeds ranged from 268 to 367 μg/g. Although HGA was present in body fluids of healthy cograzing horses, MCPA‐conjugates were not detectable, in contrast to the AM horse. Therefore, increasing concentrations of MCPA‐conjugates are supposed to be linked with the onset of AM and both parameters seem to indicate the clinical stage of disease. However, detection of HGA in body fluids of cograzing horses might be a promising step in preventing the disease.