In vitro effects of 3% hypertonic saline and 20% mannitol on canine whole blood coagulation and platelet function.

BACKGROUND: Hyperosmolar therapy, using either mannitol or hypertonic saline (HTS), is considered the treatment of choice for intracranial hypertension. However, hyperosmolar agents may impair coagulation and platelet function, limiting their use in patients at risk for hemorrhage. Despite this, studies evaluating the effects of mannitol compared to other hyperosmolar agents in dogs are largely lacking. The aim of this study was to compare the in vitro effects on global hemostasis and platelet function of 20% mannitol and 3% HTS on canine blood. METHODS: Citrated whole blood from 15 healthy dogs was diluted with 0.9% saline, 20% mannitol and 3% HTS in ratios of 1:16 and 1:8. Rotational thromboelastometry (ROTEM) was used to assess clotting time (CT), clot formation time (CFT) and maximal clot firmness (MCF) following extrinsic activation (Ex-tem) and after platelet inhibition (Fib-tem). A platelet function analyzer (PFA-100) was used to assess closure time (Ct(PFA)). RESULTS: No significant differences were observed between untreated whole blood and samples diluted with saline. Samples diluted with both mannitol and HTS were hypocoagulable compared to untreated whole blood samples. At a dilution of 1:16, no significant differences were found between any measured parameter in samples diluted with saline compared to mannitol or HTS. At a 1:8 dilution, Ct(PFA) was prolonged in samples diluted with mannitol and HTS compared to saline, and Ct(PFA) was prolonged more with mannitol than HTS. Ex-tem CT was increased at a 1:8 dilution with mannitol compared to HTS. Ex-tem CFT was prolonged at a 1:8 dilution with both agents compared to saline, and was prolonged more with mannitol than HTS. Ex-tem MCF was reduced at a 1:8 dilution with both agents compared to saline. DISCUSSION AND CONCLUSIONS: Data in this study indicate that both mannitol and HTS affect canine platelet function and whole blood coagulation in vitro in a dose-dependent fashion. The most pronounced effects were observed after high dilutions with mannitol, which impaired platelet aggregation, clot formation time, clot strength, and fibrin formation significantly more than HTS. Further in vivo studies are necessary before recommendations can be made.

Platelet-rich fibrin constructs elute higher concentrations of transforming growth factor-ß1 and increase tendon cell proliferation over time when compared to blood clots: a comparative in vitro analysis.

OBJECTIVE: To compare the concentration of a representative growth factor (transforming growth factor-beta [TGF-ß]1) eluted from a platelet-rich fibrin matrix (PRFMatrix), a platelet-rich fibrin membrane (PRFMembrane), and a whole blood clot (BC) over time, and to compare the mitogenic effect of the eluents from each construct. STUDY DESIGN: In vitro study. SAMPLE POPULATION: PRFMatrix, PRFMembrane, and BC (n=4/construct/time point). METHODS: Each construct was placed in tissue culture wells containing media for 7 days. The media was collected and replenished on days 1, 3, 5, and 7 and the concentration of eluted TGF-ß1 was measured by enzyme-linked immunosorbent assay. Canine tendon cells were subjected to additional aliquots of the conditioned media and the amount of cell proliferation compared. RESULTS: The media from both PRFM (PRFMatrix and PRFMembrane) constructs contained significantly more (P≤.026) TGF-ß1 at days 1 and 3 and produced a significant increase (P≤.044) in cell proliferation at all time points compared with the BC. The PRFMembrane media contained significantly more (P≤.05) TGF-ß1 at days 1 and 3 and produced a significant increase (P≤.002) in cell proliferation at all time points compared with the PRFMatrix. CONCLUSIONS: Both PRFM constructs are comprised of a dense fibrin scaffold that contains increased concentrations of TGF-ß1 and are capable of increasing tendon cell proliferation over time when compared with a BC. CLINICAL RELEVANCE: The sustained increase in growth factor availability in PRFM constructs may be beneficial in the healing of biologically compromised tissues.

Comparison of native and citrated whole blood samples for rapid thromboelastography in Beagles.

OBJECTIVE: To examine the extent to which rapid thromboelastography (r-TEG) could decrease the testing time in comparison with that required for kaolin-activated thromboelastography (TEG), and to compare 2 types of blood samples (ie, native and citrated whole blood [WB]), for determining r-TEG values in healthy dogs. DESIGN: Prospective observational study. SETTING: University teaching hospital. ANIMALS: Sixteen healthy Beagles. INTERVENTIONS: Kaolin-activated TEG test using citrated WB samples and r-TEG test using native and citrated WB samples were performed in 16 dogs. At 60 minutes after the initial blood sampling, further samples were collected from a subset of 6 dogs in the same manner to evaluate intraindividual repeatability of r-TEG. MEASUREMENTS AND MAIN RESULTS: The mean time to maximum amplitude (MA) for r-TEG with native and citrated WB samples was recorded as 1313.9 ± 250.9 seconds and 1351.3 ± 264.6 seconds (mean ± SD), respectively, and 1779.9 ± 197.0 seconds for kaolin-activated TEG. Coefficients of variation with native and citrated WB samples for r-TEG values, TEG-activated clotting time, clot formation time, α angle, and MA, were determined to be 13.4% versus 18.8%, 11.1% versus 16.6%, 4.2% versus 5.1%, and 10.0% versus 10.0%, respectively. Intraindividual variations were lower for native WB samples than for citrated WB samples. CONCLUSIONS: The r-TEG test significantly decreased the mean time to MA compared with the kaolin-activated TEG test. In addition, native WB samples showed lower coefficients of variation and intraindividual variation than citrated WB samples in r-TEG analysis; this suggests that native WB samples can provide more consistent results. Therefore, the r-TEG method using native WB samples is recommended for assessment of dogs' hemostatic status when an early diagnosis is required.

Thromboelastographic evaluation of hemostatic function in dogs with disseminated intravascular coagulation.

BACKGROUND: There is considerable variation in the coagulation profile of dogs with disseminated intravascular coagulation (DIC), making it difficult to assess overall hemostatic function. OBJECTIVES: To characterize the overall hemostatic state in dogs with DIC, by use of tissue factor-activated thromboelastography (TF-TEG), and to determine whether there is an association between hemostasis and outcome. ANIMALS: 50 dogs with DIC. METHODS: Dogs admitted to the intensive care units, with an underlying disease known to predispose to DIC, were prospectively assessed with TF-TEG. Citrated blood samples were collected daily during hospitalization and an extended coagulation panel and TF-TEG were performed. Diagnosis of DIC was based on expert opinion. RESULTS: Hemostatic dysfunction was observed on the TF-TEG profile in 33/50 of the dogs, of which 22/50 were hypercoagulable and 11/50 were hypocoagulable based on the TF-TEG G value alone. There were significant differences in k, alpha, and MA values (P < .0001) among hypo-, normo-, and hypercoagulable dogs. There was a significant difference in case fatality rate between hypo- (64%) and hypercoagulable (32%) dogs (relative risk = 2.38; P= .04). Dogs that died had significantly lower antithrombin activity (P= .03) and higher d-dimer concentration (P= .03) than survivors. CONCLUSIONS: The most common overall hemostatic abnormality in dogs diagnosed with DIC was hypercoagulability, and there was significant difference in survival between hyper- and hypocoagulable dogs. The results suggest TF-TEG is valuable in the assessment of hemostatic function in dogs diagnosed with DIC.

Blood coagulation times in the European brown hare (Lepus europaeus).

BACKGROUND: Many causes of mortality in the European brown hare, such as bacterial and viral infections, anticoagulant poisoning, and trauma, may result in hemorrhage. There are, however, no reference values concerning blood clotting in this species. OBJECTIVES: The aim of this study was to determine reference values for blood coagulation times and related parameters in healthy European brown hares. METHODS: Blood samples from 30 clinically healthy adult hares (15 males and 15 females) were obtained. Hares were physically restrained for blood collection from the cephalic vein into tubes containing citrate and EDTA. RESULTS: Mean +/- SD were obtained for thrombin time (TT) (13.97 +/- 1.37 seconds), prothrombin time (PT) (13.32 +/- 2.15 seconds), activated partial thromboplastin time (APTT) (16.73 +/- 1.86 seconds), fibrinogen concentration (2.98 +/- 1.06 g/L), and platelet count (355.28 +/- 128.73 x 10(9)/L). CONCLUSIONS: Reference values for blood coagulation times and other parameters associated with blood clotting will be useful in the laboratory evaluation of hemorrhage in the European brown hare.

Coagulation profiles in dogs with congenital portosystemic shunts before and after surgical attenuation.

BACKGROUND: Serious postoperative hemorrhage has been reported in dogs after closure of congenital portosystemic shunts (CPS). HYPOTHESIS: In dogs with portosystemic shunting, low coagulation factor activity is responsible for coagulopathy, which can cause complications after surgery. ANIMALS: Thirty-four dogs with CPS and 39 healthy dogs. METHODS: In a prospective study, coagulation times, platelet count, and the activity of 8 coagulation factors were measured in dogs before and after surgical shunt attenuation and in 31 healthy dogs. The effect of abdominal surgery on hemostasis was determined at ovariectomy in 8 healthy dogs. RESULTS: Dogs with CPS had lower platelet counts, lower activity of factors II, V, VII, and X, and increased factor VIII and activated partial thromboplastin time (APTT) compared to healthy dogs. After surgical attenuation, dogs with CPS had decreased platelet counts and activity of factors I, II, V, VII, IX, X, and XI and a prolonged prothrombin time (PT). Ovariectomy resulted in decreased activity of factors VII and X. Six weeks after surgery, portosystemic shunting persisted in 9 of 30 dogs, with no improvement of hemostatic values. CPS dogs without shunting had improved coagulation times and increased activity of factors II, V, VII, and X. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs with CPS have lower activity of clotting factors compared to healthy dogs, resulting in a prolonged APTT. Surgical attenuation of the shunt results in increased abnormalities in coagulation times and factors immediately after surgery. Hemostasis is normalized after complete recovery of shunting after attenuation, in contrast to dogs with persistent shunting.

Clotting profiles and selected hematology of captive Speke's gazelles (Gazella spekei).

Manual restraint and jugular venipuncture were used to obtain blood for hematology and coagulation tests for 18 captive Speke's gazelles (Gazella spekei). The hematocrit and hemoglobin values were slightly higher in Speke's gazelles than in domestic ruminants. The Speke's gazelles had a mean prothrombin time of 15.1 sec and a mean activated partial thromboplastin time of 24.2 sec. The pregnant female Speke's gazelles had shorter activated partial thromboplastin times than the males, but the difference was not significant. Ideally, prothrombin times and activated partial thromboplastin times would be compared to a healthy conspecific during a suspected bleeding crisis. Baseline prothrombin and activated partial thromboplastin times are presented here for Speke's gazelles because clotting times for exotic hoofstock are quite limited.

Determination of normal values using an automated coagulation timer for activated coagulation time and its application in dogs with hemophilia.

The present study was performed to determine normal values for the Medtronic HemoTec automated activated coagulation time (ACT) analyzer (Medtronic HemoTec Inc, Parker, CO, distributed in Switzerland by Convergenza AG, Vaduz, Liechtenstein), and to evaluate its ability to detect dogs with hemophilia. ACT was measured in 43 healthy dogs presented to the Companion Animal Hospital, University of Bern, Bern, Switzerland, with the Medtronic HemoTec ACT analyzer to determine normal values. The mean +/- 2 standard deviations (SDs) of the values obtained was defined as the normal range. ACT was measured 8-10 times on the same day in 6 dogs to determine repeatability. ACT also was measured in 11 dogs with hemophilia and compared with a conventional visual ACT measurement test and with the activated partial thromboplastin time (APTT). ACT values of the 43 dogs used to determine normal values ranged from 66.5 to 97.0 seconds (mean, 79.3 seconds; SD, 7.35 seconds; median, 78.5 seconds). A range of 64-95 seconds (mean +/- 2 SDs) was defined as the normal range for the tested device. Repeatability was poor (r = 0.256). ACT values measured with the automated device did not correlate with ACT values measured with a conventional visual test or with APTT Sensitivity of the test was 90.9%, specificity was 98.0%, and accuracy was 96.7%. Variability in the test results was large and may lead to incorrect results. The automated measurement device was not superior to the conventional visual method in evaluating dogs with hemophilia.

Changes in coagulation and fibrinolysis in horses during exercise.

Changes in clotting time (CT) and fibrinolytic activity (FA) were evaluated in 6 mature, female horses during exercise. Two trials were performed on consecutive days, using a randomized crossover design. Each mare was assigned to either an exercise trial or a control trial on the first day, and to the alternate trial 24 hours later. Mares exercised for 20 minutes on a treadmill at an elevation of 2 degrees and a velocity of 5 m/s. Venous blood samples were collected immediately before exercise, at 4, 8, 12, 16 and 20 minutes during exercise, and 15 minutes after cessation of exercise. Blood was placed into plain glass tubes for determination of CT, and into chilled, citrated tubes for determination of FA, plasminogen/plasmin complex activity (PLG), one-stage prothrombin time (OSPT), activated partial thromboplastin time (APTT), and antithrombin-III (AT-III) activity. There were significant differences (P less than 0.05) between the control and exercise groups for CT, FA, and PLG. During exercise, clotting time decreased from 21.5 +/- 1.6 minutes to 9.9 +/- 1.6 minutes (mean +/- SD; P less than 0.05), without significant changes in OSPT, APTT, or AT-III. Fibrinolytic activity and PLG increased (P less than 0.05) during exercise. Changes in CT, FA, and PLG were significant at 4 minutes of exercise, remained altered until the end of exercise, and returned to baseline values by 15 minutes of recovery. Clotting time, OSPT, APTT, FA, AT-III, and PLG did not change (P greater than 0.05) during control trials.

Evaluation of a point-of-care coagulation analyzer for measurement of prothrombin time, activated partial thromboplastin time, and activated clotting time in dogs.

OBJECTIVE: To evaluate a point-of-care coagulation analyzer (PCCA) in dogs with coagulopathies and healthy dogs. ANIMALS: 27 healthy and 32 diseased dogs with and without evidence of bleeding. PROCEDURE: Prothrombin time (PT), activated partial thromboplastin time (aPTT), and activated clotting time (ACT) were determined, using a PCCA and standard methods. RESULTS: Using the PCCA, mean (+/- SD) PT of citrated whole blood (CWB) from healthy dogs was 14.5+/-1.2 seconds, whereas PT of nonanticoagulated whole blood (NAWB) was 10.4+/-0.5 seconds. Activated partial thromboplastin time using CWB was 86.4+/-6.9 seconds, whereas aPTT was 71.2+/-6.7 seconds using NAWB. Reference ranges for PT and aPTT using CWB were 12.2 to 16.8 seconds and 72.5 to 100.3 seconds, respectively. Activated clotting time in NAWB was 71+/-11.8 seconds. Agreement with standard PT and aPTT methods using citrated plasma was good (overall agreement was 93% for PT and 87.5% for aPTT in CWB). Comparing CWB by the PCCA and conventional coagulation methods using citrated plasma, sensitivity and specificity were 85.7 and 95.5% for PT and 100 and 82.9% for aPTT, respectively. Overall agreement between the PCCA using NAWB and the clinical laboratory was 73% for PT and 88% for aPTT. Using NAWB for the PCCA and citrated plasma for conventional methods, sensitivity and specificity was 85.7 and 68.4% for PT and 86.7 and 88.9% for aPTT, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The PCCA detected intrinsic, extrinsic, and common pathway abnormalities in a similar fashion to clinical laboratory tests.

Reference values for activated coagulation time in cats.

OBJECTIVE: To establish reference values for activated coagulation time (ACT) in cats by use of jugular venipuncture and direct collection of blood into ACT vacuum tubes. ANIMALS: 100 clinically normal cats that were to have elective surgery performed at a private practice. PROCEDURE: Collection of 3 blood samples for ACT measurement was attempted for each cat at the time of elective surgery: sample 1, obtained before sedation; sample 2, tube 1 of 2 consecutive samples obtained from a single venipuncture of the contralateral jugular vein after sedation with acepromazine and ketamine hydrochloride; and sample 3, tube 2 collected immediately following collection of sample 2 without removing the needle from the vein. Venipuncture quality was rated subjectively on a 3-point scale. RESULTS: Median ACT were 95 seconds for each sample group. The middle 95% of values ranged inclusively from 55 to 185 seconds (sample 1), 65 to 135 seconds (sample 2), 45 to 145 seconds (sample 3), and 55 to 165 seconds overall (samples 1, 2, and 3). Significant differences in ACT values were not detected between sample groups. Significant relationships between ACT and venipuncture quality or sex of cat were not detected. CONCLUSIONS AND CLINICAL RELEVANCE: With the ACT protocols used, clinically normal cats had ACT of < 165 seconds. The ACT in cats does not appear to be significantly affected by sex, sedation with acepromazine and ketamine, or by moderately traumatic venipunctures. These results refute widespread statements that ACT should be < 65 seconds in healthy cats. Cats with ACT repeatedly > 165 seconds should be further evaluated for hemostatic disorders.

Activated partial thromboplastin time and activated coagulation time in monitoring heparinized cats.

Cats were given heparin (IV or subcutaneously) at various dosages to determine the optimal dosage, route, and schedule for therapeutic anticoagulation. The activated coagulation time and activated partial thromboplastin time tests were used to evaluate heparin concentration. A dosage of 100 U/kg given IV produced an anticoagulation that was too brief and more potent than was desirable. Subcutaneous dosages of 250 and 375 U/kg produced therapeutically acceptable anticoagulation for 8 hours, whereas dosages of 500 and 1,000 U/kg causes an anticoagulation that was longer and more potent than was desirable. The activated coagulation time correlated well with the activated partial thromboplastin time in determining the degree of heparinization with certain limitations.

Activated coagulation time in monitoring heparinized dogs.

Heparin was administered to normal dogs either IV or subcutaneously and the anticoagulant effects were monitored by determining prolongation of activated partial thromboplastin time (APTT) and activated coagulation time (ACT). Significant correlation was found between ACT and log APTT for all heparin doses. Intravenously administered heparin was rapidly cleared, with the anticoagulant effects lasting only 2 to 3 hours. One hour after IV heparin at dose levels of 100 and 200 U/kg was given, the APTT were prolonged to 1.8 and 2.8 X base-line values, respectively, whereas the ACT were prolonged to 1.4 and 1.6 X base-line values, respectively. Subcutaneously administered heparin was rapidly absorbed, approaching maximal anticoagulation by 2 hours. Thereafter, a fairly stable plateau of anticoagulation was maintained for 4 to 14 hours depending on dose. Two hours after administration of heparin dosages of 250, 500, 750, and 1,000 U/kg, the dog's APTT values were prolonged by 1.6, 2.4, 3.1, and 3.2 X base line, respectively. Corresponding values for ACT revealed prolongations of 1.3, 1.4, 1.6, and 1.7 X base line. Considerable variation in APTT response to a given dose of heparin was noted, particularly in dogs given the higher heparin doses. Subcutaneous heparin administration at a dosage of 500 U/kg maintained an APTT target value of 1.5 to 2.5 X base line for 8 hours. On the basis of the regression line equation, ACT = -7.2 + 73.5 log APTT, ACT values corresponding to the APTT target value ranged from 92.2 to 108.5 s (1.23- to 1.45-fold base line). Monitoring anticoagulation in dogs given subcutaneous heparin is recommended at 2 hours to evaluate maximal anticoagulation effects. The inexpensiveness, technical simplicity, and linear response to heparin support the use of ACT in monitoring heparinized dogs.

The thrombocyte aggregation process in the turtle Phrynopys hilarii (Chelonia). An ultrastructural study.

This study investigates the thrombocyte aggregation process in the South American fresh water turtle (Phrynopys hilarii) using electron microscopy. Blood was taken from surgically exposed lateral neck vessels of ten turtles Phrynopys hilarii during the spring and summer seasons, when the mean temperature is 37 degrees C. Blood samples were fixed with Karnovsky solution for processing by transmission electron microscopy. The turtle thrombocytes were spindle-shaped with lobulated nuclei. Prominent vesicles and canaliculi were found throughout the cytoplasm. The cytoplasm organelles showed an agranular endoplasmatic reticulum, Golgi complex near the centrioles and scattered free ribosomes. These cells are similar to bird thrombocytes but distinct from fish and frog thrombocytes. Blood clotting time was 5 min +/- 30 sec measured by the Lee and White method. Structural alterations resulting from the aggregation process occurred after activation. Thrombocytes developed numerous filopodial projections, an increased number of vacuoles and changed from spindle to spherical shape. P. hilarii thrombocytes have different morphologic characteristics compared to other non-mammalian vertebrate cells. These cells can participate in the aggregation process, as observed in birds.

Evaluation of point-of-care tests for diagnosis of disseminated intravascular coagulation in dogs admitted to an intensive care unit.

OBJECTIVE: To evaluate the accuracy of point-of-care tests for the diagnosis of disseminated intravascular coagulation (DIC) in dogs and assess the correlation and agreement of results between point-of-care and laboratory tests in the evaluation of hemostatic function. DESIGN: Prospective case series. ANIMALS: 59 critically ill dogs (affected dogs) with clinical signs of diseases known to predispose to DIC and 52 clinically normal dogs. PROCEDURES: Accuracy of the point-of-care tests (activated clotting time [ACT], estimated platelet count and number of schizocytes from a blood smear, plasma total solids [TS] concentration, and the protamine sulfate test) was evaluated, using receiver operating characteristic curves and likelihood ratios. A strategy, using likelihood ratios to calculate a posttest probability of DIC, was tested with 65% used as a threshold for initiation of treatment. Results of laboratory tests (coagulogram and plasma antithrombin III activity) were used as the standard for comparison in each dog. RESULTS: ACT and estimated platelet count provided the best accuracy for detection of DIC. The plasma TS concentration, schizocyte number, and protamine sulfate test had poor accuracy. The strategy using post-test probability of DIC identified 12 of 16 affected dogs that had DIC. Estimated platelet count was correlated and had acceptable clinical agreement with automated platelet count (r = 0.70). The plasma TS (r = 0.28) concentration and serum albumin (r = 0.63) concentration were not accurate predictors of plasma antithrombin III activity. The ACT did not correlate with activated partial thromboplastin time (r = 0.28). CONCLUSIONS AND CLINICAL RELEVANCE: Strategic use of likelihood ratios from point-of-care tests can assist clinicians in making treatment decisions for dogs suspected to have DIC when immediate laboratory support is unavailable.

Preliminary observations on the effect of Dictyocaulus filaria on the blood clotting time of sheep.

The whole blood clotting time progressively increased from the second to the eighth week in lambs receiving a primary infection of Dictyocaulus filaria larvae. However, in vaccinated lambs it remained unaffected. Levamisole hydrochloride was ineffective when given 4 days after infection but restored the blood clotting time to near normal soon after treatment when it was given 30 days after infection.

[The "activated coagulation time (ACT)": two simple screening methods for evaluating coagulation disorders in dogs]. / Die "Activated Coagulation Time (ACT)": Zwei einfache Screening-Methoden zur Erfassung von Gerinnungsstörungen beim Hund.

The use of the activated coagulation time (ACT) for testing the intrinsic coagulation is well established among veterinary practitioners in the USA. The advantage of the ACT compared to other coagulation tests is its ease to be performed under practice conditions. The ACT may be measured manually or instrumentally. The reference range of our instrumental measurement is between 90 and 120 seconds (median 105 seconds), of the manual measurement at room temperature between 115 and 145 seconds (median 125 seconds). Advantages of the instrumental method are the smaller amount of blood (0.4 ml versus 2.0 ml) necessary to perform the test, and the smaller potential for errors by unexperienced examiners. The spread is comparable between the two methods. The most important cause of false results is poor venipuncture technique: traumatic venipuncture will trigger the coagulation cascade already during venipuncture causing an artificially shortened ACT.

Sonoclot evaluation of whole blood coagulation in healthy adult dogs.

OBJECTIVE: To establish a standard protocol for analysis of canine whole blood and generate reference intervals for healthy dogs using the Sonoclot analyzer, and to compare Sonoclot values to standard and viscoelastic coagulation tests. DESIGN: Prospective study. SETTING: Veterinary University research facility and teaching hospital. ANIMALS: Twelve healthy random source dogs and 52 healthy dogs from the general veterinary school population. INTERVENTIONS: Blood sampling for viscoelastic coagulation testing. MEASUREMENTS AND MAIN RESULTS: Blood was collected from 12 healthy adult dogs by jugular venipuncture. After a rest period at room temperature of 30, 60, or 120 minutes, 340 µL of citrated blood was added to 20 µL of 0.2 M CaCl(2) in 1 of 2 cuvette types warmed to 37° C. Cuvettes contained a magnetic stir-bar with glass beads (gbACT+) or only a magnetic stir-bar (nonACT). Reference interval samples were collected from 52 healthy adult dogs and analyzed in duplicate. The ACT, CR, and PF were not affected by duration of rest period for either cuvette type. ACT variability was decreased when using gbACT+ cuvettes (P < 0.05). In normal dogs reference intervals (mean ± 2 SD) using gbACT+ cuvettes were: ACT 56.0-154.0 seconds, CR 14.85-46.0, and PF 2.1-4.05. ACT correlated to TEG R-time, K-time, and angle, while CR correlated with all TEG parameters. Fibrinogen correlated with ACT, CR, and PF. Sonoclot did not correlate with other common coagulation tests. CONCLUSIONS: Sonoclot provides viscoelastic evaluation of canine whole blood coagulation and correlated to several TEG parameters and fibrinogen. A standard protocol and reference intervals were established.

In vitro evaluation of canine hemostasis following dilution with hydroxyethyl starch (130/0.4) via thromboelastometry.

OBJECTIVE: To assess the effects of in vitro hemodilution of canine blood with a low molecular weight hy-droxyethyl starch (HES 130/0.4) by means of thromboelastometry (TEM). DESIGN: In vitro experimental study. SETTING: University Teaching Hospital. ANIMALS: Ten healthy adult staff-owned dogs were sampled for the purposes of the study. Samples were also collected from 45 clinically normal dogs to establish thromboelastometric reference intervals. INTERVENTIONS: For each dog whole blood was collected by jugular venipuncture and placed into tubes containing 3.8% trisodium citrate and stored at 37°C. Two standard dilutions (1:4 and 1:10) from canine blood specimens were prepared with HES 130/0.4 and saline 0.9%. MEASUREMENTS AND MAIN RESULTS: The effects of dilution were studied by TEM. No statistically significant differences were observed between the TEM values of the control samples and the samples diluted with saline solution (ratio 1:10 and 1:4). In contrast, hypocoagulability was observed in the samples treated with HES 130/0.4, with more severe effects at 1:4 dilution than at 1:10 dilution. The 1:4 dilution with HES 130/0.4 produced marked alterations: CT (P = 0.035) and CFT (P = 0.0007) on the ex-TEM profile, CT (P = 0.0005) and ML (P = 0.0017) on the fib-TEM profile and CFT (P = 0.0043) on the in-TEM, were all significantly increased (P < 0.05), whereas MCF (P = 0.0042) and alpha angle (P = 0.002) on the in-TEM and MCF (P = 0.011) and alpha angle (P = 0.001) ex-TEM profiles and MCF (P = 0.0001) on the fib-TEM profile were significantly decreased (P < 0.05). CONCLUSIONS: Dilution of canine blood with HES 130/0.4 results in a thromboelastometric pattern consistent with hypocoagulation and this effect appears to result from a dose-dependent alteration in fibrinogen concentration and inhibition of platelet function. As this is an in vitro study, further in vivo investigations are necessary to confirm the results.

Changes of platelet function and blood coagulation during short-term storage of CPDA-1-stabilised ovine blood.

The objective of this study was to detect the influence of short-term storage on the haemostatic function in whole citrated ovine blood at different storage temperatures. Ovine blood was collected in a commercial transfer bag system containing CPDA-1 and stored on a wobbler at room (20-25 °C; n=5) or refrigerator temperature (4 °C; n=5). The following analyses were performed initially and after 1, 2, 3, 4, 5, 6, 8, 12, 24, 48 and 72 h of storage: platelet count and (spontaneous) aggregates, agonist-induced platelet aggregation with two methods (impedance aggregometry, turbidimetric method), prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen concentration and resonance thrombography. Platelet count remained stable at room temperature, whereas a significant decrease was detected after 48 h storage at 4 °C. The latter was associated with the formation of a high percentage of platelet aggregates (50-60%) after 5h storage. Decrease in platelet aggregation was significantly more pronounced when blood was stored at 4°C. The plasmatic coagulation tests were stable within the observation period. Results indicate that platelet count and aggregability of CPDA-1-stabilised ovine blood is better preserved at room temperature and provides adequate haemostatic function for ex vivo experiments for one working day. Functional loss and high percentage of platelets within aggregates which were observed in ovine blood stored at refrigerator temperature have to be considered in blood transfusion in sheep.