Semester of Graduation

Spring 2020

Degree

Master of Science (MS)

Department

Veterinary Clinical Sciences

Document Type

Thesis

Abstract

A broad knowledge base on veterinary transfusion medicine has been developed in common mammalian species, but little is known in reptiles. The goals of this thesis were to establish laboratory methods to assess the application of blood transfusions in reptiles. Morphological and physiological differences in reptiles, including their erythrocytes, hamper direct adaption of mammalian laboratory methods.

Quantification of percentage hemolysis is a crucial tool for quality assessment of stored red blood cell products. We evaluated measurement of the required analytes for this calculation (supernatant and total hemoglobin) with different methods in blood from American alligators and found that the use of point-of care analyzers is a feasible alternative to the time-consuming reference method at clinically relevant levels of medium hemolysis. Cross-matching is an important step during donor selection, but little is known on compatibilities between reptiles and the procedure is not routinely performed. We evaluated a modified cross-matching protocol that only requires a minimum amount of EDTA blood to overcome frequent challenges of small reptiles. Intraspecies incompatibilities have been found in 6/7 tested bearded dragons, and interspecies incompatibilities have been found between all lizards tested against a snake and an American alligator.

Studies on post-transfusion red blood cell survival (RCS) are a powerful diagnostic tool to assess different variables that can bias the benefits of blood transfusion. A non-radioactive method (biotinylation) has become more popular in mammals. We established this method in reptiles and succesfully performed multi-density labeling using 0.03 and 0.45 pg NHS-biotin/RBC with 0.3 µg of streptavidin-phycoerythin per 1 x 106 RBC as secondary label. Then, we used the biotinylation technique to assess the effect of blood storage and of donors on RCS in bearded dragons. We found that there is no difference in RCS between freshly transfused autologous erythrocytes and those stored for 28 days, but there was a significant difference depending on the donor. Longest RCS was found after autologous transfusions (erythrocyte lifespan 236 days), whereas homologous RCS survivial was shorter (erythrocyte lifespan: 65 days), but alligator erythrocytes (heterologous transfusion) were only recovered for 1-3 days post transfusion (erythrocyte lifespan 2.7 days).

Committee Chair

Nevarez; Javier G

DOI

10.31390/gradschool_theses.5140

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