Evaluation of contact activation of citrated equine whole blood during storage and effects of contact activation on results of recalcification-initiated thromboelastometry

Am J Vet Res. 2015 Feb;76(2):122-8. doi: 10.2460/ajvr.76.2.122.

Abstract

Objective: To evaluate the degree of activation of the contact pathway in citrated equine whole blood over holding times ≤ 30 minutes and assess effects of contact activation on recalcification-initiated thromboelastometry.

Animals: 11 healthy adult mixed-breed horses.

Procedures: Blood was collected by atraumatic jugular venipuncture into prewarmed evacuated siliconized glass tubes containing citrate anticoagulant and held at 37°C for ≤ 30 minutes. Thromboelastometry was performed with an in vitro viscoelasticity (thromboelastometry) monitoring system. Factor XII and factor XI procoagulant activities were determined in contemporaneously collected platelet-poor plasma samples by assessing changes in turbidity for 1 hour at approximately 25°C, with clotting times calculated by fitting a line to the steepest segment of the absorbance curve and determining its intersection with baseline. Effect of holding time on thromboelastometry parameters and plasma enzyme activity was evaluated by repeated-measures ANOVA on ranks. Association of procoagulant activities with coagulation time was determined by Spearman rank-order correlation analysis.

Results: Thromboelastometry parameters (coagulation time, clot formation time, α angle, and maximum clot firmness) reflected significant increases in coagulability during the holding period. Factor XII and factor XI procoagulant activities were significantly increased at 30 minutes, compared with 2 or 10 minutes (indicating contact activation of samples), and had significant negative correlation with coagulation time.

Conclusions and clinical relevance: Ex vivo activation of the contact system in equine whole blood was evident, suggesting that recalcification of blood in the absence of a trigger is not an acceptable method of assessing the hemostatic system in horses.

MeSH terms

  • Animals
  • Blood Coagulation Factors / pharmacology*
  • Citrates
  • Female
  • Horses
  • Male
  • Specimen Handling / veterinary
  • Thrombelastography / veterinary*

Substances

  • Blood Coagulation Factors
  • Citrates