A Method of Isolating Trypanosomes from Blood

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Abstract
Treatment of heparinized or citrated blood with Bacto-Phytohemagglutinin (Difco, Code 0528) makes possible the removal of almost all mature erythrocytes by low speed centrifugation, leaving most of the trypanosomes in the supernate. The flagellates can be concentrated by centrifugation at higher speed and then washed, if desirable, by resuspension and centrifugation in appropriate solutions. Blood forms of Trypanosoma cruzi, isolated and concentrated by this technic, were normal with respect to morphology, virulence for mice, and infectivity for tissue culture cells. Similarly, blood forms of T. gambiense, isolated from rat blood, were normal in appearance, and virulence for rats was maintained in freshly isolated flagellates as well as those which were preserved at -65[degree]C for 8 months. Suspensions of T. gambiense which were essentially free of all cellular components of blood, such as leucocytes and platelets, were prepared by treating the mixture of flagellates and non-agglutinated blood cells with antiserum from a rabbit immunized by serial inoculations of suspensions of non-agglutinable cells; these inocula were prepared by treating rat blood with phytohemagglutinin. This method has been used to isolate large numbers of trypanosomes from blood and also to concentrate into a drop or two of fluid the flagellates from relatively large volumes of blood taken from animals with very light infections. The technic may be useful in isolating extracellular blood parasites for physiologic studies or antigen preparation.

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