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Protocol
Establishing a protocol for thromboelastography in sea turtles
  1. Ashley Barratclough1,
  2. Rita Hanel2,
  3. Nicole I Stacy3,
  4. Laura K Ruterbories2,
  5. Emily Christiansen4,5 and
  6. Craig A Harms5
  1. 1 Department of Large Animal Clinical Sciences, University of Florida, College of Veterinary Medicine, Gainesville, Florida, USA
  2. 2 Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA
  3. 3 Department of Diagnostic, Comparative and Population Medicine, University of Florida, College of Veterinary Medicine, Gainesville, Florida, USA
  4. 4 North Carolina Aquariums, Raleigh, North Carolina, USA
  5. 5 Department of Clinical Sciences and Environmental Medicine Consortium, College of Veterinary Medicine, Center for Marine Sciences and Technology, North Carolina State University, Morehead City, North Carolina, USA
  1. Correspondence to Dr Ashley Barratclough; barratclough{at}ufl.edu

Abstract

Thromboelastography (TEG) provides a global evaluation of haemostasis. This diagnostic test is widely used in mammals but has not previously been performed in reptiles, mainly due to the limited availability of taxon-specific reagents. The objective of this pilot study was to establish a protocol to perform TEG in sea turtles. Pooled citrated plasma, stored at −80°C, from four green turtles (Chelonia mydas) was assayed on a TEG 5000. Several initiators were evaluated: kaolin (n=2), RapidTEG (n=2), fresh (n=2) and frozen (n=6) thromboplastin extracted from pooled brain tissue from several chelonian species, human recombinant tissue factor at 1:100 (n=1), Reptilase (n=2), and rabbit thromboplastin (n=1). Both fresh and frozen chelonian thromboplastin were superior in producing quantifiable TEG reaction time compared with all other reagents. These findings are consistent with the lack of an intrinsic pathway in turtles and confirmed a lack of coagulation in the turtle samples in response to mammalian thromboplastin. A TEG protocol was subsequently established for harvested species-specific frozen thromboplastin. The frozen thromboplastin reagent remained stable after one year of storage at −80°C. The developed protocol will be useful as a basis for future studies that aim to understand the pathophysiology of haemostatic disorders in various stranding conditions of sea turtles.

  • thromboelastography
  • sea turtle
  • coagulation

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Footnotes

  • Contributors All coauthors contributed significantly to this work. CAH and AB had the original idea to the study. NIS developed the idea significantly and created the plan for the research project. RH was instrumental in the execution of the project and providing the equipment, and LKR performed the methodology. EC assisted with sampling from the study. All authors assisted with data analysis and writing the research report. All authors have agreed to this final submitted version. Everyone listed as an author fulfils all three of the ICMJE guidelines for authorship.

  • Funding The authors have not declared a specific grant for this research from any funding agency in the public, commercial or not-for-profit sectors.

  • Competing interests None declared.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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