Thromboelastography in equine medicine: Technique and use in clinical research

Thromboelastography (TEG) is a viscoelastic, whole blood‐based assay that integrates information from both the cellular and soluble components of coagulation, providing a global evaluation of the haemostatic system. This contrasts with the conventional coagulation assays (i.e. platelet count, prothrombin time [PT], activated partial thromboplastin time [aPTT] and fibrinogen concentration [FIB]), which only provide information about one component (e.g. clotting factors in the case of PT and aPTT) of the haemostatic process, requiring the combination of several assays for a complete evaluation of haemostasis. Thromboelastography is an old technology that has been used in human medicine for over 50 years. However, it is relatively new in veterinary medicine and has only been applied to horses in the last 5 years. Clinical applications in human medicine include diagnosis and monitoring of coagulopathies. Currently, extensive research is being carried out to expand the use of TEG in dogs and cats. Therefore, it is expected that the use of this technique will also further expand in horses in the near future. To date, the available studies in the equine species have evaluated TEG in healthy horses, horses with gastrointestinal disease, septic foals, horses with exercise‐induced pulmonary haemorrhage (EIPH) and a filly with Glanzmann's thrombasthenia. The main objective of this review is to introduce the TEG technique to equine clinicians, providing information on how the TEG functions, blood sample collection and processing, variables measured and their interpretations, normal reference values and areas of potential clinical application.     

Plasma concentrations of endothelin-like immunoreactivity in healthy horses and horses with naturally acquired gastrointestinal tract disorders

OBJECTIVE: To compare plasma endothelin (ET)- like immunoreactivity between healthy horses and those with naturally acquired gastrointestinal tract disorders. ANIMALS: 29 healthy horses and 142 horses with gastrointestinal tract disorders. PROCEDURE: Blood samples were collected from healthy horses and from horses with gastrointestinal tract disorders prior to treatment. Magnitude and duration of abnormal clinical signs were recorded, and clinical variables were assessed via thorough physical examinations. Plasma concentrations of ET-like immunoreactivity were measured by use of a radioimmunoassay for human endothelin-1, and CBC and plasma biochemical analyses were performed. RESULTS: Plasma ET-like immunoreactivity concentration was significantly increased in horses with gastrointestinal tract disorders, compared with healthy horses. Median plasma concentration of ET-like immunoreactivity was 1.80 pg/ml (range, 1.09 to 3.2 pg/ml) in healthy horses. Plasma ET-like immunoreactivity was greatest in horses with strangulating large-intestinal obstruction (median, 10.02 pg/ml; range, 3.8 to 22.62 pg/ml), peritonitis (9.19 pg/ml; 789 to 25.83 pg/ml), and enterocolitis (8.89 pg/mI; 6.30 to 18.36 pg/ml). Concentration of ET-like immunoreactivity was significantly associated with survival, PCV, and duration of signs of pain. However, correlations for associations with PCV and duration of pain were low. CONCLUSIONS AND CLINICAL RELEVANCE: Horses with gastrointestinal tract disorders have increased plasma concentrations of ET-like immunoreactivity, compared with healthy horses. The greatest values were detected in horses with large-intestinal strangulating obstructions, peritonitis, and enterocolitis. This suggests a potential involvement of ET in the pathogenesis of certain gastrointestinal tract disorders in horses.     

Oesophageal electrocardiography in healthy horses

Reasons for performing study: In human medicine, oesophageal electrocardiography (ECG) is a well‐established technique that magnifies P waves with respect to the QRS complex. Objectives: To investigate the feasibility of oesophageal ECG recording in horses and its ability to produce larger P waves compared with base‐apex and unipolar recordings. Methods: Bipolar and unipolar ECG were performed using oesophageal and surface electrodes. Oesophageal ECG was obtained from 6 different recording configurations at different oesophageal depths. Amplitudes of P, Q, R, S and T waves were measured from 3 different cardiac cycles for each recording configuration and depth. Results: Oesophageal ECG was feasible in all horses. For all oesophageal recording configurations, significantly larger P waves were recorded from a depth that equalled ‘height of the withers + 10 cm’ (HW₊₁₀) than from any other depth. P/QRS_magn, the ratio between the P wave and QRS complex magnitudes, was largest for intraoesophageal recordings with an interelectrode distance of 10 cm, at HW₊₁₀, where it was significantly larger than base‐apex and unipolar recordings. Base‐apex recording resulted in significantly smaller P waves than all other recording configurations and significantly smaller P/QRS_magn ratios than all other recording configurations except one combined oesophageal‐surface recording (E/S_low). Conclusions: Oesophageal ECG recording is feasible in horses and effective in magnifying P wave amplitude. Potential relevance: The procedure is promising for diagnosis of supraventricular tachydysrhythmias and might be used in electrophysiological studies and for cardiac pacing.     

Putative intestinal hyperammonaemia in horses: 36 cases

Reasons for performing the study: Intestinal hyperammonaemia (HA) has been infrequently reported in individual horses; however, there have been no studies describing clinical and laboratory data as well as short‐ and long‐term outcome in a larger number of cases. Objectives: To describe clinical and laboratory data and short‐ and long‐term outcome in a large group of horses with intestinal HA. Methods: Multi‐centred, retrospective study; case records of horses with HA were reviewed and any horse with a clinical or post mortem diagnosis of intestinal HA was included. Hyperammonaemia was defined as a blood ammonium (NH₄⁺) concentration ≥60 µmol/l and horses with a diagnosis of primary hepatic disease were excluded. Relevant data were recorded and, if appropriate, data from survivors were compared to nonsurvivors to identify potential prognostic indicators. Results: Thirty‐six cases, 26 mature horses and 10 foals with intestinal HA were identified. Case histories included diarrhoea, colic and neurological signs and the most common clinical diagnosis was colitis and/or enteritis. The most common clinical and laboratory abnormalities included tachycardia, increased packed cell volume, hyperlactataemia and hyperglycaemia. Fourteen horses (39%) survived to discharge; NH₄⁺ concentration on admission was the only parameter significantly associated with survival. All surviving horses and foals for which follow‐up information was available recovered completely and returned to their intended use without further complications. Conclusions and potential relevance: Intestinal HA occurs in mature horses and foals and can be associated with severe clinical and laboratory abnormalities; further studies are required to investigate predisposing factors and delineate possible differences in aetiologies.     

Fungal sinusitis resulting in suspected trigeminal neuropathy as a cause of headshaking in five horses

Primary fungal sinusitis was identified in 5 horses displaying signs of headshaking. All 5 horses had fungal plaques adhered to the infraorbital canal (IOC). Headshaking signs were exhibited by 3 horses prior to treatment and 2 horses after treatment. Standing computed tomography (CT) identified erosion of the IOC in the 2 cases in which it was performed. Fungal culture and PCR identified 3 species of fungi, Rhizomucor pusillus, Scedosporium apiospermum and Aspergillus nidulans which have not previously been described as a cause of sinusitis in horses. Surgical debridement followed by topical antifungal therapy was used in all 5 horses. Recurrence of the fungal plaques in 4 horses necessitated further treatment. The headshaking signs and nasal discharge resolved in 3 horses allowing a return to their previous use. Two horses developed persistent headshaking signs despite multiple treatments. Primary fungal sinusitis should be considered as a cause of headshaking signs in horses, due to a suspected trigeminal neuropathy. Computed tomography is valuable in identifying erosion of the IOC which is not identified with conventional radiography. Three out of the 5 cases were treated successfully but permanent resolution of the fungal infection is difficult to achieve once the bone overlying the infraorbital nerve has been eroded.     

Pharmacokinetics of amikacin in plasma and selected body fluids of healthy horses after a single intravenous dose

Reasons for performing study: No studies have determined the pharmacokinetics of low‐dose amikacin in the mature horse. Objectives: To determine if a single i.v. dose of amikacin (10 mg/kg bwt) will reach therapeutic concentrations in plasma, synovial, peritoneal and interstitial fluid of mature horses (n = 6). Methods: Drug concentrations of amikacin were measured across time in mature horses (n = 6); plasma, synovial, peritoneal and interstitial fluid were collected after a single i.v. dose of amikacin (10 mg/kg bwt). Results: The mean ± s.d. of selected parameters were: extrapolated plasma concentration of amikacin at time zero 144 ± 21.8 µg/ml; extrapolated plasma concentration for the elimination phase 67.8 ± 7.44 µg/ml, area under the curve 139 ± 34.0 µg*h/ml, elimination half‐life 1.34 ± 0.408 h, total body clearance 1.25 ± 0.281 ml/min/kg bwt; and mean residence time (MRT) 1.81 ± 0.561 h. At 24 h, the plasma concentration of amikacin for all horses was below the minimum detectable concentration for the assay. Selected parameters in synovial and peritoneal fluid were maximum concentration (C_max) 19.7 ± 7.14 µg/ml and 21.4 ± 4.39 µg/ml and time to maximum concentration 65 ± 12.2 min and 115 ± 12.2 min, respectively. Amikacin in the interstitial fluid reached a mean peak concentration of 12.7 ± 5.34 µg/ml and after 24 h the mean concentration was 3.31 ± 1.69 µg/ml. Based on a minimal inhibitory concentration (MIC) of 4 µg/ml, the mean C_max : MIC ratio was 16.9 ± 1.80 in plasma, 4.95 ± 1.78 in synovial fluid, 5.36 ± 1.10 in peritoneal fluid and 3.18 ± 1.33 in interstitial fluid. Conclusions: Amikacin dosed at 10 mg/kg bwt i.v. once a day in mature horses is anticipated to be effective for treatment of infection caused by most Gram‐negative bacteria. Potential relevance: Low dose amikacin (10 mg/kg bwt) administered once a day in mature horses may be efficacious against susceptible microorganisms.     

Ethyl pyruvate diminishes the inflammatory response to lipopolysaccharide infusion in horses

Reasons for performing the study: Endotoxaemia contributes to morbidity and mortality in horses with colic due to inflammatory cascade activation. Effective therapeutic interventions are limited for these horses. Ethyl pyruvate (EP), an anti‐inflammatory agent that alters the expression of proinflammatory cytokines, improved survival and organ function in sepsis and gastrointestinal injury in rodents and swine. Therapeutic efficacy of EP is unknown in endotoxaemic horses. Objectives: Determine the effects of EP on signs of endotoxaemia and expression of proinflammatory cytokines following administration of lipopolysaccharide (LPS) in horses. Methods: Horses received 30 ng/kg bwt LPS in saline to induce signs of endotoxaemia. Next, horses received lactated Ringer's solution (LRS), (n = 6), 150 mg/kg bwt EP in LRS, (n = 6), or 1.1 mg/kg bwt flunixin meglumine (FM), (n = 6). Controls received saline followed by LRS (n = 6). Physical examinations, behaviour pain scores and blood for clinical pathological testing and gene expression were obtained at predetermined intervals for 24 h. Results: Lipopolysaccharide infusion produced clinical and clinicopathological signs of endotoxaemia and increased expression of tumour necrosis factor alpha (TNFα), interleukin 6 (IL‐6) and IL‐8 (P<0.001) compared with controls. Leucopenia and neutropenia occurred in all horses that received LPS. Horses treated with EP and FM had significantly (P<0.0001) reduced pain scores compared with horses receiving LPS followed by LRS. Flunixin meglumine was significantly more effective at ameliorating fever compared with EP. Both EP and FM significantly diminished TNFα expression. Ethyl pyruvate significantly decreased, but FM significantly increased, IL‐6 expression. Neither EP nor FM altered IL‐8 expression. Conclusions and potential relevance: Ethyl pyruvate administered following LPS diminished the clinical effects of endotoxaemia and decreased proinflammatory gene expression in horses. Ethyl pyruvate suppressed expression of proinflammatory cytokines better than FM. However, FM was a superior anti‐pyretic compared with EP. Ethyl pyruvate may have therapeutic applications in endotoxaemic horses.     

Association between hypercoagulability and decreased survival in horses with ischemic or inflammatory gastrointestinal disease

Background: Coagulopathies are common in horses with ischemic or inflammatory gastrointestinal (GI) disturbances. There is indirect evidence suggesting that early stages of these diseases are characterized by hypercoagulability (HC). Hypothesis/Objectives: HC, assessed via thromboelastography (TEG), is common in horses with ischemic or inflammatory GI diseases. The degree of HC is correlated with nonsurvival and thrombotic complications. Animals: Thirty client‐owned horses with ischemic or inflammatory GI disease, 30 client‐owned horses with nonischemic or inflammatory GI disease, and 30 healthy horses (control group). Methods: Prospective, observational clinical study. TEG profiles of 30 horses with ischemic or inflammatory GI disease were obtained on admission and 48 hours after admission, and these were compared with profiles from 30 horses with nonischemic or inflammatory GI disease and 30 healthy controls. Prothrombin time (PT), activated partial thromboplastin time (aPTT), antithrombin activity (AT), and D‐Dimer concentrations were also determined in horses with GI disease. Results: Horses with ischemic or inflammatory GI disease had shorter R times compared with healthy horses (14.8 ± 8.3 versus 22.8 ± 12 minute; P= .011). However, changes were subtle and TEG profiles did not resembled those obtained from animals or humans presumed to be hypercoagulable. Although conventional coagulation testing supported the presence of HC (decreased AT and increased D‐Dimer concentrations), TEG and coagulation abnormalities were rarely found in the same horses and the methods were not statistically related. Conclusions and Clinical Importance: There is evidence of HC in horses with GI disease but techniques for diagnoses require refinement.     

Preliminary investigation of concurrent administration of phenylbutazone and romifidine in healthy horses

ObjectiveTo characterize the cardiorespiratory and electrocardiographic effects of the combined administration of phenylbutazone and romifidine.Study designProspective four-period, four-treatment, blinded, randomized, crossover trial.AnimalsFive, healthy, mixed breed horses.MethodsPrior to treatment administration, a catheter was introduced into the intra-thoracic cranial vena cava via the jugular vein and a subcutaneously located carotid artery was catheterised. All treatments were administered intravenously (IV) and consisted of saline placebo (PLC), phenylbutazone (PBZ, 4.4 mg kg^?1) romifidine (ROM, 80 μg kg^?1) and a combination of phenylbutazone (4.4 mg kg^?1) and romifidine (80 μg kg^?1). There was at least a 1 week washout period between treatments. Heart rate (HR), respiratory rate (f_R), systolic (SAP), diastolic (DAP) and mean (MAP) arterial pressures and central venous pressure (CVP) were recorded for baseline (prior to drug administration) and at 5 minute intervals thereafter for 30 minutes. Electrocardiographic abnormalities were recorded. Data were analyzed by anova.ResultsFor the cardiovascular variables there were no statistically significant (p > 0.05) differences between horses treated with ROM and PBZ_ROM. Statistically significant (p < 0.05) differences only occurred between treatments with romifidine (ROM and PBZ_ROM) and without romifidine (PLC and PBZ). Within treatments, for ROM, changes over time were statistically significant (p < 0.05) for HR, SAP, DAP, MAP and CVP. For PBZ_ROM, changes over time were statistically significant (p < 0.05) for CVP. Sino-atrial and atrio-ventricular blocks occurred in horses treated with ROM and PBZ_ROM.Conclusions and clinical relevanceThe combined IV administration of phenylbutazone and romifidine had no statistically significant effect on cardiorespiratory variables. These limited data suggest no evidence why both agents should not be included in a preoperative medication protocol for healthy horses but do not exclude the possibility of interactions occurring in a larger population.     

Oesophageal disorders in horses: Retrospective study of 39 cases

Oesophageal diseases are less frequently reported in equine veterinary practice in comparison with other alimentary pathologies. This paper describes the prevalence, age, breed and sex distribution, the duration of clinical presentation before admission, the diagnosis, aetiology, treatment course, complications, length of hospitalisation, and the short‐term and long‐term survival of 39 cases with equine oesophageal disorders at a referral clinic. Prevalence of oesophageal disorders was significantly higher in the group of horses up to 4 years of age in comparison with other age groups. The most common oesophageal disorder was simple obstruction represented by 22/39 cases (56.41%). The common complications of the oesophageal disorder were oesophagitis (16/39, 41.02%) and aspiration pneumonia (11/39, 28.20%); 31/39 (79.49%) of all oesophageal cases survived up to clinical discharge. Long‐term survival was 65.78% in this group of cases.