A technique for isolating heterophils from blood of orange-winged Amazon parrots (Amazona amazonica amazonica)

Blood samples from adult orange-winged Amazon parrots (Amazona amazonica amazonica) were collected to develop a rapid and efficient technique for isolating pure populations of morphologically intact and functional heterophils. In addition, normal haematological parameters for the orange-winged Amazon parrots (n=20) were established and found to be within the reported range of other Amazon species. Heterophil isolation (n=16) was maximally achieved by concentrating the white blood cells with 3% hetastarch prior to running the sample through a polysucrose and disodium diatrizoate centrifugation. The isolation procedure yielded an average heterophil recovery of 61.9%, the purity exceeded 92.6% and viability was 98.9%. Cell integrity was evaluated utilising flow cytometry, cytochemistries, and light microscopy. In this study, isolated heterophils exhibited morphological and cytochemical characteristics similar to whole blood heterophils and were phagocytically active. The results of this study demonstrate that an intact, viable and functionally active heterophil population can successfully be isolated from relatively small volumes of blood and that these cell populations can be further studied employing in vitro bioassaying techniques.