Comparison of Jugular vs. Saphenous Blood Samples, Intrarater and In-Between Device Reliability of Clinically Used ROTEM S Parameters in Dogs.

Rotational Thromboelastometry (ROTEM) allows for the global assessment of hemostasis in whole blood samples. Preanalytical and analytical factors may influence test results, and data about the reliability and reproducibility of lyophilized ROTEM tests are scarce. Therefore, the objective of this study was to evaluate the influence of blood collection site on ROTEM S parameters and to assess intrarater and in-between device variability. A total of thirty, healthy, staff-owned dogs were included. Blood collection and ROTEM analysis were performed by trained staff according to a standardized protocol. Extrinsically activated (tissue factor; Ex-TEM S), with the addition of cytochalasin for platelet inhibition (Fib-TEM S), and intrinsically activated (In-TEM) analyses were performed. Analysis of our data showed significant variability for various Ex-TEM S and Fib-TEM S parameters from different collection sites and intrarater and in-between device measurements. We conclude that serial monitoring with ROTEM should be performed on the same device, with blood always taken from the same collection site using a standardized blood sampling technique. While In-TEM S, apart from maximum lysis, showed very stable and reliable results, we suggest interpreting especially clotting and clot formation parameters from Ex-TEM S and Fib-TEM S tests with caution and using duplicate measurements to detect outliers and to prevent initiation of incorrect therapies.

Evaluation of the Effect of Storage Time on ROTEM S® Parameters in Healthy and Ill Dogs.

Viscoelastic testing as a bedside test to assess global haemostasis has gained popularity in the past decade, with rotational thromboelastometry (ROTEM) and thromboelastography (TEG) being the two commonly used devices. TEG studies suggest analysis 30 min after blood sampling. However, the reproducibility of results over time for ROTEM analysis using lyophilized samples in dogs has not been established. In this study, we investigated the influence of time on viscoelastic testing, using 33 healthy staff-/client-owned dogs for blood sampling and repeated measurements of ROTEM tracings at three different time points after blood collection. Additionally, a group of 21 hospitalized patients with suspected coagulation disorders were included to investigate whether stability over time was comparable between healthy and ill dogs. We demonstrated a significant difference of ROTEM tracings over time, with a tendency towards hypocoagulability over time. These changes do have a clinical relevance as they exceed reference intervals and could therefore lead to erroneous conclusions about a patient's coagulation status. Therefore, time-specific reference intervals are proposed and presented in this publication.

Characterization of Acute Traumatic Coagulopathy in Cats and Association with Clinicopathological Parameters at Presentation.

OBJECTIVE: This study aimed to document rotational thromboelastometric (ROTEM) characteristics of traumatized cats and to investigate associations between clinicopathological parameters and acute traumatic coagulopathy (ATC). A secondary goal was to determine the relevance of autoheparinization in injured cats. STUDY DESIGN: Cats presenting with acute (<12 hours) trauma were eligible. Cats were allocated to the ATC group (≥2 hypocoagulable parameters) or non-ATC group (≤1 hypocoagulable parameter) based on ROTEM analysis. Clinicopathological parameters were compared between groups and regression was used to find variables associated with ATC. Heparinase-modified ROTEM (HepTEM) was used to assess for heparin effects in a subgroup. RESULTS: Fifty-three cats were included, and the incidence of ATC was 15%. Prolongation of both intrinsic and extrinsic clotting times (CT) was the most frequently altered ROTEM variable in the ATC group, but CTInTEM-prolongation also occurred in 47% of non-ATC cats. The incidence of autoheparinization, defined as concurrent CTInTEM prolongation and CTInTEM:HepTEM ratio >1.1, was 41% and was observed in both cats with and without ATC. None of the evaluated clinicopathological parameters were different between groups or associated with ATC. CONCLUSION: Acute traumatic coagulopathy in cats is mainly characterized by prolonged CT. No relationship between clinicopathological variables and ATC was identified and prediction of ATC based on these variables was not possible. While autoheparinization is important in cats, it is not the sole cause for ATC.

Whole blood platelet impedance aggregometry with the ROTEM platelet device: comparison of 2 anticoagulants and storage times for the establishment of canine reference intervals.

The ROTEM platelet device, a point-of-care whole blood platelet impedance aggregometer, is an add-on to the rotational thromboelastometry ROTEM delta device. The latter has been validated in dogs. We examined whether canine whole blood is suited for analysis with the ROTEM platelet device using adenosine-5'-diphosphate (ADP) and arachidonic acid (ARA) as agonists for platelet activation, and if there are significant differences between sample storage times and anticoagulants used. Subsequently, we determined canine reference intervals (RIs) for the ROTEM platelet device for ADP and ARA. In a pilot study, we examined whole blood from 7 dogs after 15-min and 60-min storage of lithium-heparinized samples and 40-min and 80-min storage of hirudinized samples. Statistical analysis showed no significant differences between ROTEM platelet device results for both ADP and ARA in lithium-heparin and hirudin anticoagulated canine whole blood. Lithium-heparinized blood samples analyzed after 15-min storage had the lowest coefficient of variation. RIs were determined for heparinized whole blood samples from 49 dogs after 15 min of storage.

Coagulation Status in Dogs Naturally Infected with Angiostrongylus vasorum.

Angiostrongylus vasorum infection has been associated with coagulopathies including hyperfibrinolysis. We compared coagulation status including thromboelastometry (ROTEM) parameters in dogs naturally infected with A. vasorum versus healthy dogs to determine clinicopathological parameters associated with bleeding, hypocoagulopathy, and hyperfibrinolysis. Clinical signs, white blood cell count, platelet count, hematocrit, plasmatic coagulation tests (PT, aPTT, fibrinogen concentration), D-dimer, and ROTEM S parameters (Ex-tem, In-tem, Fib-tem, Ap-tem) were analysed and compared between bleeding, nonbleeding, and control dogs and between hypo- and normocoagulable animals. Clinical signs of bleeding were present in 6/9 (67%) hypocoagulable and 1/9 (11%) normocoagulable dogs. PT, fibrinogen concentration, and several ROTEM parameters were significantly different between hypocoagulable and normocoagulabe A. vasorum infected dogs. Hyperfibrinolysis was identified in 44% of infected dogs and was significantly more common in bleeding and hypocoagulable dogs. Hyperfibrinolysis was significantly associated with low MCFFib-tem but not with low fibrinogen concentration or increased D-dimers. CFTEx-tem > 248 swas 100% sensitive and 89% specific to predict hyperfibrinolysis. Hyperfibrinolysis, hypocoagulability and bleeding are common in A. vasorum infected dogs. Only Ex-tem and Fib-tem parameters and potentially PT were associated with bleeding or hypocoagulability. Ex-tem analysis enables detection of bleeding, hypocoagulability and hyperfibrinolysis within minutes.

Prevalence of Acute Traumatic Coagulopathy in Acutely Traumatized Dogs and Association with Clinical and Laboratory Parameters at Presentation.

OBJECTIVE: The aim of this study was to determine the prevalence of acute traumatic coagulopathy (ATC) and identify associated clinical and laboratory parameters including rotational thromboelastometry. STUDY DESIGN: Dogs presenting within 6 hours after trauma were allocated to the ATC or non-ATC group based on thromboelastometry analysis (ex-tem S, in-tem S, fib-tem S). ATC was defined as ≥2 hypocoagulable parameters in 1 profile and ≥ 1 hypocoagulable parameter in an additional profile. Parameters used were ex-tem and in-tem clotting time (CT), clot formation time (CFT), maximum clot firmness (MCF), maximum lysis and fib-tem MCF. Clinical and laboratory parameters at presentation, animal trauma triage (ATT) score, transfusion requirement and outcome were compared. Logistic regression was used to identify independent factors associated with ATC. RESULTS: Eleven of 33 dogs presented with ATC and showed ex-tem CT and CFT prolongation and reduced MCF amplitude in all profiles (all p < 0.001). pH (p = 0.043) and potassium concentration (p = 0.022) were significantly lower and bleeding (p = 0.027) and plasma transfusions (p = 0.001) more common in dogs with ATC. Time after trauma (p = 0.040) and Animal Trauma Triage score (p = 0.038, including haematocrit as confounding factor) were associated with the presence of ATC. CONCLUSION: Acute traumatic coagulopathy is more common in traumatized dogs than previously reported. Acute traumatic coagulopathy was associated with acidosis, Animal trauma triage score, time after trauma and higher transfusion needs. Coagulation abnormalities include ex-tem CT and CFT prolongations and decreased clot strength.

Determination of reference intervals for single vial rotational thromboelastometry (ROTEM) parameters and correlation with plasmatic coagulation times in 49 clinically healthy dogs.

The objectives of this prospective study were determination of reference intervals (RI) for rotational thromboelastometry (ROTEM) parameters in single use reagents and to evaluate correlations between plasmatic coagulation times and ROTEM parameters. Blood was sampled from a jugular vein in 49 client-owned healthy dogs and ex-tem S, in-tem S, fib-tem S and ap-tem S parameters, prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen, haematology, blood chemistry and venous blood gas analysis was performed. Determination of RI was performed using Excel add-in Reference Value Advisor and correlations between PT, aPTT and fibrinogen with selected ROTEM parameters were determined by Spearman correlation. Ex-tem S maximum clot firmness (MCF) RI are smaller compared to RI in people and liquid ex-tem in dogs while maximum lysis was comparable to those in people but smaller than previously reported in dogs. A strong correlation was found between fibrinogen measured by Clauss and fib-tem S and in-tem S MCF (r = 0.541, P < .001 and r = 0.610, P < .001, respectively). PT showed a significant but moderate correlation with ex-tem S CT (r = 0.340, P = .030), in-tem S CFT (r = 0.433, P = .003), fib-tem S CT (r = 0.426, P = .009) and ap-tem S CT (r = 0.354, P = .015) while aPTT was not significantly correlated with any of the evaluated parameters. In conclusion, this study provides single use reagent ROTEM parameter RIs that are different from RI determined with liquid reagents. Significant correlations between fibrinogen concentrations measured by Clauss and clot firmness of fib-tem S and in-tem S profiles and between PT and clotting times of all reagents were identified.

Serial Evaluation of Haemostasis Following Acute Trauma Using Rotational Thromboelastometry in Cats.

OBJECTIVE: The aim of this study was to describe coagulation abnormalities and incidence of acute traumatic coagulopathy (ATC) in traumatized cats over the first 24 hours after admission. STUDY DESIGN: This was a prospective observational study at the university teaching hospital including 26 cats with acute (<5 hours) trauma. Blood was sampled for rotational thromboelastometry (ROTEM) parameters at presentation and 6 hours/24 hours thereafter. Rotational thromboelastometry tracings were defined as hypo- or hypercoagulable if ≥ 2 of the following parameters were above or below institutional reference intervals: clotting time, clot formation time (CFT), maximum clot firmness, maximum lysis or maximum clot elasticity. Hypocoagulability at presentation was defined as ATC. Injury severity scores, treatment and survival to hospital discharge were retrieved from patient records. RESULTS: The incidence of ATC was 15% and the most common ROTEM abnormalities in cats with ATC were clotting time and CFT prolongation in both extrinsic and intrinsic ROTEM profiles. After 24 hours, compared with presentation, significantly more cats were hypercoagulable (p = 0.047) and none of the cats showed hypocoagulopathy. Cats with ATC received significantly more blood transfusions (p = 0.008). CONCLUSION: The incidence of ATC in cats is higher than previously reported. Clotting time and CFT prolongations seem to be more common than hyperfibrinolysis and 53% of the cats became hypercoagulable within 24 hours. While the clinical relevance of ATC in cats needs to be investigated, cats diagnosed with ATC required significantly more blood transfusions.

Reference values for rotational thromboelastometry (ROTEM) in clinically healthy cats.

OBJECTIVE: To establish reference intervals for rotational thromboelastometry (ROTEM) using feline blood. DESIGN: Prospective study. SETTING: University teaching hospital. ANIMALS: Twenty-three clinically healthy cats between 1 and 15 years. INTERVENTION: For each cat, whole blood was collected via jugular or medial saphenous venipuncture, and blood was placed into a serum tube, a tube containing potassium-EDTA, and tubes containing 3.2% sodium citrate. The tubes were maintained at 37°C for a maximum of 30 minutes before coagulation testing. MEASUREMENTS AND MAIN RESULTS: ROTEM tests included the EXTEM, INTEM, FIBTEM, and APTEM assays. In addition, prothrombin time, activated partial thromboplastin time, thrombin time, and fibrinogen concentration (Clauss method) were analyzed for each cat. Reference intervals for ROTEM were calculated using the 2.5-97.5th percentile for each parameter, and correlation with the standard coagulation profile was performed. Compared to people, clinically healthy cats had similar values for the EXTEM and INTEM assays, but had lower plasma fibrinogen concentrations (0.9-2.2 g/L), resulting in weaker maximum clot firmness (MCF, 3-10 mm) in the FIBTEM test. In 18 cats, maximum lysis (ML) values in the APTEM test were higher than in the EXTEM test, which seems unlikely to have occurred in the presence of aprotinin. It is possible that the observed high maximum lysis values were due to clot retraction rather than true clot lysis. Further studies will be required to test this hypothesis. CONCLUSIONS: Cats have a weaker clot in the FIBTEM test, but have a similar clot strength to human blood in the other ROTEM assays, which may be due to a stronger contribution of platelets compared to that found in people. In cats, careful interpretation of the results to diagnose hyperfibrinolysis is advised, especially with the APTEM test, until further data are available.

The use of strain, strain rate, and displacement by 2D speckle tracking for assessment of systolic left ventricular function in goats: applicability and influence of general anesthesia.

BACKGROUND: Assessment of left ventricular (LV) systolic function can be achieved by conventional echocardiographic methods, but quantification of contractility, regional myocardial function, and ventricular synchrony is challenging. The goal of this study was to investigate the applicability of two-dimensional speckle tracking (2DST) to characterize segmental and global wall motion for assessment of LV function and LV synchrony in healthy goats. We aimed to describe the techniques, report normal values of a variety of 2DST indices, and determine the influence of general anesthesia. METHODS: Prospective study on 22 healthy female Saanen goats (3.7 ± 1.1 y, 60.2 ± 10.5 kg [mean ± SD]). All goats underwent two transthoracic echocardiographic examinations, the first standing and unsedated and the second 7.4 ± 3.5 days later during isoflurane anesthesia and positioned in sternal recumbency. Data analyses were performed offline, blinded, and in random order. Left ventricular longitudinal, radial and circumferential strain and strain rate as well as longitudinal and radial displacement were measured using 2DST methods. Summary statistics were generated and differences of 2DST variables between myocardial segments and treatments (i.e., awake vs. anesthetized) were assessed statistically (alpha level=0.05). RESULTS: Echocardiographic analyses by 2DST were feasible in all goats and at both time points. Longitudinal systolic strain, strain rate and displacement followed a gradient from apex to base. Absolute systolic strain was generally lower and strain rate was higher in awake goats compared to anesthetized goats. Circumferential and radial indices did not consistently follow a segmental pattern. Generally, peak strain occurred later in anesthetized goats compared to awake goats. General anesthesia did not significantly influence LV synchrony. CONCLUSIONS: 2SDT is a valid method for non-invasive characterization of LV wall motion in awake and anesthetized goats. The results of this study add to the understanding of LV mechanical function, aid in the diagnosis of global and segmental LV systolic dysfunction, and will be useful for future cardiovascular studies in this species. However, effects of anesthesia and species-specific characteristics should be considered when goats are used as animal models for human disease.

Comparison of pulmonary artery and transpulmonary thermodilution cardiac output measurements in unsedated newborn calves.

OBJECTIVE: To compare the agreement, repeatability and trending ability of transpulmonary thermodilution (TPTD) and pulmonary artery thermodilution (PATD) cardiac output (Q˙t) measurements in unsedated newborn calves. STUDY DESIGN: Prospective experimental trial. ANIMALS: Eight newborn calves weighing a median (range) of 53 (46-59) kg. METHODS: Pulmonary and femoral artery thermodilution catheters were placed under local anaesthesia. A total of 382 PATD and TPTD Q˙t measurements were performed simultaneously. Cardiac output was influenced by intravenous doxapram and theophylline in a randomized crossover fashion. Bland-Altman analysis for multiple comparisons, concordance and polar plots were used to assess TPTD against PATD. RESULTS: Median (range) cardiac index values measured with PATD and TPTD were 197 mL kg(-1) minute(-1) (74-335 mL kg(-1) minute(-1)) and 196 mL kg(-1) minute(-1) (59-395 mL kg(-1) minute(-1)), respec-tively. A small mean bias of -3 mL kg(-1) minute(-1) with limits of agreement (LOA) of -64 to 58 mL kg(-1) minute(-1) and a percentage error of 31% were found. Eighty-two mean values were calculated. This reduced the LOA to -50 to 41 mL kg(-1) minute(-1) with a similar small bias and a percentage error of 23%. Mean TPTD tracked changes in Q˙t compared with mean PATD with 90% concordance, a mean polar angle of 6° and radial LOA of 43°, indicating marginal trending ability. Keeping the femoral artery catheter patent and obtaining acceptable measurements were very challenging because the calves were not used to being restrained. Calf movement had less influence on PATD. CONCLUSIONS AND CLINICAL RELEVANCE: We recommend that PATD remains the reference method to measure Q˙t in unsedated newborn calves. However, the robust results of the evaluation of the less invasive TPTD technique warrants further evaluation taking into account the difficulties reported in this study.