Effect of hemolysis on plasma protein levels and plasma electrophoresis in birds

Protein electrophoresis is recognized as a reliable diagnostic tool for birds even though results are seldom pathognomonic. Unfortunately, this technique is underused in avian medicine because many factors interfere with electrophoresis patterns; hemolysis is one of these factors and is often associated with improper specimen handling. In human laboratory medicine, hemolysis is a known interference factor that can lead to erroneous results. Published data on the influence of hemolysis on protein electrophoresis in birds is currently restricted to a single study in Psittacidae. The aim of this study was to further investigate this effect and to analyze potential interspecific differences. Blood samples were drawn from 28 Black Kites (Milvus migrans) and 19 Bar-headed Geese (Anser indicus) and separated into two aliquots. One aliquot was dipped into liquid nitrogen for 5 sec in order to cause freeze-thawing hemolysis before centrifugation. Total plasma protein concentration, plasma hemoglobin concentration, and plasma protein electrophoresis patterns were determined for both hemolyzed and nonhemolyzed samples. In both species, hemolysis resulted in falsely elevated total plasma protein concentration. In Bar-headed Geese, hemolysis caused a rise in the gamma fraction. In Black Kites, this rise involved not only the gamma fraction but also the beta fraction, stressing the potential for species-related differences. In both species, the effects of hemolysis mimicked a chronic inflammatory condition with resulting antigenic stimulation.