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Abstract

OBJECTIVE

To compare the effects of open-tube blood sampling with previously investigated blood sampling methods via evacuated tube on thromboelastography variables for blood samples from dogs.

ANIMALS

10 healthy Beagles from the research colony owned by the Clinic of Small Animal Internal Medicine, University Veterinary of Medicine, Vienna, were used.

PROCEDURES

In this prospective study, blood was sampled from each dog serially into citrate solution–containing tubes via 20-gauge needle. One evacuated tube was filled from a jugular vein via the evacuated tube port, and the second tube was opened and filled by catching blood flowing through the needle from a lateral saphenous vein. Venipuncture quality was scored with a previously described method. Thromboelastography was performed for each sample.

RESULTS

Inferential statistics used with the Wilcoxon signed rank test showed significant differences in reaction time (R) of 3.43 ± 0.84 minutes versus 4.53 ± 0.62 minutes (mean ± SD) between evacuated tube assisted and open-tube sampling, respectively. No other significant differences were identified.

CLINICAL RELEVANCE

The sampling methods compared have a small but significant effect on R in thromboelastographic analysis for blood samples from healthy dogs. Shear stress by vacuum sampling seems to accelerate coagulation in jugular blood samples harvested by evacuated tube, resulting in a shortened R. Results suggested that the open-tube method avoids shear stress induced activation of coagulation and is an appropriate sampling method for thromboelastography when used within a standardized protocol.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To determine the effects of leukoreduction on N-methylhistamine (NMH; a stable histamine metabolite) concentration in units of canine whole blood during storage and incubation at room temperature (approx 22 °C) to simulate temperature conditions during transfusion.

ANIMALS

8 healthy adult Walker Hounds.

PROCEDURES

A standard unit of blood (450 mL) was obtained from each dog twice, with at least 28 days between donations. Blood units collected from 4 dogs during the first donation underwent leukoreduction, whereas the blood units collected from the other 4 dogs did not undergo leukoreduction, prior to storage at 4 °C. The alternate treatment was applied to blood units collected during the second donation. A sample from each unit was obtained for determination of plasma NMH concentration the day of donation (before and after leukoreduction when applicable) and before and after incubation at room temperature for 5 hours on days 14 and 28 of storage.

RESULTS

Units that underwent leukoreduction had substantially lower leukocyte and platelet counts than nonleukoreduced units. Plasma NMH concentration increased immediately after leukoreduction but did not change significantly during the subsequent 28 days of storage, nor did it differ between units that did and did not undergo leukoreduction.

CONCLUSIONS AND CLINICAL RELEVANCE

Leukoreduction and simulated transfusion temperature did not affect the histamine load in units of canine whole blood during the first 28 days of storage. Further research is necessary to determine whether histamine contributes to the development and severity of blood transfusion reactions in dogs.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To assess clotting times, coagulation factor activities, sterility, and thromboelastographic parameters of liquid plasma (LP), thawed fresh frozen plasma (FFP-T), and 2 novel formulations of freeze-dried plasma (FDP) stored refrigerated over 35 days.

SAMPLE

6 units of canine LP and FFP-T from a commercial animal blood bank and 5 units each of 2 formulations of canine FDP.

PROCEDURES

Prothrombin time; activated partial thromboplastin time; activities of coagulation factors II, V, VII, VIII, IX, X, XI, and XII; and thromboelastographic parameters were determined for each product on days 0 (baseline), 3, 7, 14, 21, 28, and 35. For each day, a sample of each product was also submitted for aerobic bacterial culture.

RESULTS

Small changes in coagulation factor activities and mild increased time to initial clot formation in LP and FFP-T were noted over the 35-day storage period. Activities of factor VIII in FDP1 and factor XII in FDP2 were < 50% at baseline but varied throughout. Compared with FFP-T, time to initial clot formation was increased and clot strength was preserved or increased for the FDPs throughout the study. One FDP had decreased pH, compared with other products. No plasma product yielded bacterial growth.

CONCLUSIONS AND CLINICAL RELEVANCE

Liquid plasma and FFP-T would be reasonable to use when stored refrigerated for up to 35 days. Both FDP products showed variability in coagulation factor activities. Studies investigating the usefulness of these plasma products (FDPs) in dogs and the variable days of refrigerated storage (all products) are warranted. (Am J Vet Res 2020;81:964–972)

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To examine the effects of imidazoline and nonimidazoline α-adrenergic agents on aggregation of feline platelets.

SAMPLE

Blood samples from 12 healthy adult cats.

PROCEDURES

In 7 experiments, the effects of 23 imidazoline and nonimidazoline α-adrenoceptor agonists or antagonists on aggregation and antiaggregation of feline platelets were determined via a turbidimetric method. Collagen and ADP were used to initiate aggregation.

RESULTS

Platelet aggregation was not induced by α-adrenoceptor agonists alone. Adrenaline and noradrenaline induced a dose-dependent potentiation of ADP- or collagen-induced aggregation. Oxymetazoline and xylometazoline also induced a small potentiation of ADP-stimulated aggregation, but other α-adrenoceptor agonists did not induce potentiation. The α2-adrenoceptor antagonists and certain imidazoline α-adrenergic agents including phentolamine, yohimbine, atipamezole, clonidine, medetomidine, and dexmedetomidine inhibited adrenaline-potentiated aggregation induced by ADP or collagen in a dose-dependent manner. The imidazoline compound antazoline inhibited adrenaline-potentiated aggregation in a dose-dependent manner. Conversely, α1-adrenoceptor antagonists and nonimidazoline α-adrenergic agents including xylazine and prazosin were ineffective or less effective for inhibiting adrenaline-potentiated aggregation. Moxonidine also was ineffective for inhibiting adrenaline-potentiated aggregation induced by collagen. Medetomidine and xylazine did not reverse the inhibitory effect of atipamezole and yohimbine on adrenaline-potentiated aggregation.

CONCLUSIONS AND CLINICAL RELEVANCE

Adrenaline-potentiated aggregation of feline platelets may be mediated by α2-adrenoceptors, whereas imidazoline agents may inhibit in vitro platelet aggregation via imidazoline receptors. Imidazoline α-adrenergic agents may have clinical use for conditions in which there is platelet reactivity to adrenaline. Xylazine, medetomidine, and dexmedetomidine may be used clinically in cats with minimal concerns for adverse effects on platelet function.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To determine whether passage of whole blood through a microaggregate filter by use of a syringe pump would damage canine erythrocytes.

SAMPLE

Blood samples obtained from 8 healthy client-owned dogs.

PROCEDURES

Whole blood was passed through a standard microaggregate filter by use of a syringe pump at 3 standard administration rates (12.5, 25, and 50 mL/h). Prefilter and postfilter blood samples were collected at the beginning and end of a simulated transfusion. Variables measured at each time point included erythrocyte osmotic fragility, mean corpuscular fragility, RBC count, hemoglobin concentration, RBC distribution width, and RBC morphology. In-line pressure when blood passed through the microaggregate filter was measured continuously throughout the simulated transfusion. After the simulated transfusion was completed, filters were visually analyzed by use of scanning electron microscopy.

RESULTS

Regardless of administration rate, there was no significant difference in mean corpuscular fragility, RBC count, hemoglobin concentration, or RBC distribution width between prefilter and postfilter samples. Additionally, there were no differences in in-line pressure during the simulated transfusion among administration rates. Echinocytes were the erythrocyte morphological abnormality most commonly observed at the end of the transfusion at administration rates of 12.5 and 25 mL/h.

CONCLUSIONS AND CLINICAL RELEVANCE

Results suggested that regardless of the administration rate, the microaggregate filter did not alter fragility of canine RBCs, but may have altered the morphology. It appeared that the microaggregate filter would not contribute to substantial RBC damage for transfusions performed with a syringe pump.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To evaluate coagulation factors in units of leukoreduced (LR) and nonleukoreduced (non-LR) canine fresh-frozen plasma (cFFP).

ANIMALS

8 healthy research dogs.

PROCEDURES

In a crossover study, dogs were randomly assigned to 1 of 2 groups from which blood was collected and either did or did not undergo leukoreduction. After a recovery period of ≥ 28 days, the dogs were switched between protocols. After each collection, blood samples were centrifuged, and cFFP was stored frozen for later comparative analysis of coagulation factors, antithrombin, and protein C activities (reported as comparative percentages of the corresponding activities determined in a canine pooled plasma standard); prothrombin and activated partial thromboplastin times; and fibrinogen concentration.

RESULTS

There were no significant differences detected between results for LR cFFP, compared with those for non-LR cFFP.

CONCLUSIONS AND CLINICAL RELEVANCE

Although there was variation among residual activities of coagulation factors in LR and non-LR cFFP, the variations and differences were considered unlikely to impact the efficacy of LR cFFP transfused for coagulation factor replacement in dogs. However, owing to the small sample size and high variability of results in the present study, additional research with a larger sample size is required for definitive conclusions on the effects of leukoreduction on coagulation factors in cFFP and to develop treatment guidelines for LR cFFP use in dogs with congenital and acquired coagulopathies.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To evaluate stability of coagulation factors in canine plasma obtained by use of plasmapheresis and stored over a 36-month period.

SAMPLE

Canine plasma obtained by use of plasmapheresis acquired from a commercial blood bank.

PROCEDURES

Coagulation testing for fibrinogen concentration and activity of factors II, V, VII, VIII, and IX and von Willebrand factor was performed on canine plasma obtained by use of plasmapheresis. Samples were obtained for testing at 6-month intervals from plasma stored for up to 36 months.

RESULTS

A simple mixed linear regression model was created for each analysis. Median value for the fibrinogen concentration was > 150 mg/dL for all time points, except at 467, 650, and 1,015 days of storage. Median value for factor VIII was > 70% only at 650 days. Median value for factor V was > 50% through 650 days. Median value for factors VII and X was > 50% through 833 days, and median value for factors II and VII was > 50% through 1,015 days. Median value for von Willebrand factor was > 50% for the entire study (1,198 days). Median value for factor X was always < 50%.

CONCLUSIONS AND CLINICAL RELEVANCE

Coagulation factors degraded over time at variable rates, and all labile factors remained at > 50% activity for longer than 1 year. Plasma collected by plasmapheresis potentially offers prolonged life span of some clotting factors. Plasmapheresis is an acceptable form of canine plasma collection for transfusion purposes, and further studies should be performed to determine all of its benefits.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To assess pharmacokinetics of tranexamic acid (TXA) in dogs and assess antifibrinolytic properties of TXA in canine blood by use of a thromboelastography-based in vitro model of hyperfibrinolysis.

ANIMALS 6 healthy adult dogs.

PROCEDURES Dogs received each of 4 TXA treatments (10 mg/kg, IV; 20 mg/kg, IV; approx 15 mg/kg, PO; and approx 20 mg/kg, PO) in a randomized crossover-design study. Blood samples were collected at baseline (time 0; immediately prior to drug administration) and predetermined time points afterward for pharmacokinetic analysis and pharmacodynamic (thromboelastography) analysis by use of an in vitro hyperfibrinolysis model.

RESULTS Maximum amplitude (MA [representing maximum clot strength]) significantly increased from baseline at all time points for all treatments. The MA was lower at 360 minutes for the 10-mg/kg IV treatment than for other treatments. Percentage of clot lysis 30 minutes after MA was detected was significantly decreased from baseline at all time points for all treatments; at 360 minutes, this value was higher for the 10-mg/kg IV treatment than for other treatments and higher for the 20-mg/kg IV treatment than for the 20-mg/kg PO treatment. Maximum plasma TXA concentrations were dose dependent. At 20 mg/kg, IV, plasma TXA concentrations briefly exceeded concentrations suggested for complete inhibition of fibrinolysis. Oral drug administration resulted in a later peak antifibrinolytic effect than did IV administration.

CONCLUSIONS AND CLINICAL RELEVANCE Administration of TXA improved clot strength and decreased fibrinolysis in blood samples from healthy dogs in an in vitro hyperfibrinolysis model. Further research is needed to determine clinical effects of TXA in dogs with hyperfibrinolysis.

Full access
in American Journal of Veterinary Research

Abstract

OBJECTIVE To validate that dogs become hypocoagulable following rattlesnake envenomation and to determine whether thromboelastographic abnormalities are correlated with envenomation severity for dogs bitten by rattlesnakes native to southern California.

ANIMALS 14 dogs with observed or suspected rattlesnake envenomation (envenomated dogs) and 10 healthy control dogs.

PROCEDURES For each dog, a citrate-anticoagulated blood sample underwent kaolin-activated thromboelastography. For each envenomated dog, a snakebite severity score was assigned on the basis of clinical findings, and prothrombin time, activated partial thromboplastin time, and platelet count were determined when the attending clinician deemed it necessary and owner finances allowed.

RESULTS For 12 of 14 envenomated dogs, the thromboelastographically determined clot strength was below the 25th percentile for the clot strength of control dogs, which was indicative of a hypocoagulable state. No envenomated dog had thromboelastographic results indicative of a hypercoagulable state. One envenomated dog had a prolonged prothrombin time, but the activated partial thromboplastin time and all thromboelastographic variables were within the respective reference ranges for that dog. Seven of 13 envenomated dogs were thrombocytopenic (platelet count, ≤ 170,000 platelets/μL). Snakebite severity score was negatively correlated with platelet count but was not correlated with any thromboelastographic variable.

CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that dogs generally become hypocoagulable following rattlesnake envenomation. Thromboelastography might provide an objective measure of the coagulation status of envenomated dogs and aid in the identification of dogs that are in a hypocoagulable state and in need of antivenin treatment prior to the onset of progressive clinical signs.

Full access
in American Journal of Veterinary Research