Abstract

Hemolysis frequently results from pre-analytical factors and is a commonly encountered interference in serum or plasma chemistry data.^1,2 Although a mild degree of hemolysis is considered to cause negligible changes in chemistry results, moderate to severe hemolysis has the potential to alter particularly electrolytes, minerals, and protein electrophoresis to a degree that may affect clinical decisions.^1-4 Little is known about erythrocyte chemistry in nondomestic species, but understanding methodology - and species-specific effects of hemolysis resulting from various analytical methodologies and erythrocyte contents, respectively, is needed for accurate interpretation of chemistry results. The objective of this study was to evaluate the effects of moderate to severe hemolysis on plasma electrolytes, minerals, and protein electrophoresis in loggerhead sea turtles (Caretta caretta; Cc). Paired non-hemolyzed plasma and whole blood samples previously collected in lithium heparin from 16 Cc were frozen at -80C. After thawing, the non-hemolyzed plasma sample was divided into one aliquot of native plasma and one aliquot to which lysed whole blood was added to create moderate to severe hemolysis. Chemistry analysis was performed using methods previously described.^5,6 Potassium, magnesium, phosphorus, total protein, alpha-1-, alpha-2-, and beta-globulins were significantly higher and albumin:globulin ratio significantly lower in hemolyzed compared to non-hemolyzed plasma. Potassium was the most affected analyte with up to 4-fold increase in hemolyzed samples. No significant differences were observed for sodium, chloride, calcium, pre-albumin, albumin, and gamma-globulins. These results demonstrate the importance of considering potential interfering effects of hemolysis in moderately to severely hemolyzed plasma from loggerhead sea turtles when interpreting chemistry data and emphasize the necessity to document visual hemolysis as component of the chemistry report. This study highlights the need to understand methodology - and species-specific effects of hemolysis in plasma chemistry.

Acknowledgements

The authors wish to thank veterinary and rehabilitation staff at the Loggerhead Marinelife Center and the staff of University of Miami Avian & Wildlife Laboratory.

* Presenting author

Literature Cited

1.  Stockham SL, Scott MA. 2008. Fundamentals of Veterinary Clinical Pathology. Ames, IA: Blackwell Publishing Professional. 908.

2.  Yucel D, Dlava K. 1992. Effect of in vitro hemolysis on 25 common biochemical tests. Clin Chem. 38:575–577.

3.  Benson KG, Paul-Murphy J, MacWilliams P. 1999. Effects of hemolysis on plasma electrolyte and chemistry values in the common green iguana (Iguana iguana). J Zoo Wildl Med. 38(3):413–415.

4.  Hawkins MG, Kass PH, Zinkl JG, Tell LA. 2006. Comparison of biochemical values in serum and plasma, fresh and frozen plasma, and hemolyzed samples from orange‐winged Amazon parrots (Amazona amazonica). Vet Clin Pathol. 35(2):219–225.

5.  Deem SL, Norton TM, Mitchell M, Segars AL, Alleman AR, Cray C, Poppenga RH, Dodd M, Karesh WB. 2009. Comparison of blood values in foraging, nesting, and stranded loggerhead turtles (Caretta caretta) along the coast of Georgia, USA. J Wildl Dis. 45(1):41–56.

6.  Knafo SE, Norton TM, Mitchell M, Stevenson DJ, Hyslop N, Poppenga R, Oliva M, Chen T, Cray C, Gibbs SE, et al. 2016. Health assessment of Eastern indigo snakes (Drymarchon couperi) in Georgia. J Zoo Wildl Med. 47(4):1000–1012.