Red blood cell (RBC) negative charges and resistance to linoleic acid (LNA)-induced lysis were studied in Plasmodium yoelii-infected mice and in malaria (P. falciparum)-affected individuals. RBCs from mice infected with P. yoelii showed a progressive decrease in the net surface negative charges at 24 h after infection, reaching a minimal value on day 3, followed by a second phase that was characterised by a recovery to normal levels on day 6. Resistance to linoleic acid follows similar kinetics. These alterations preceded the appearance of parasites in the peripheral blood. A similar increase in LNA-induced lysis was observed in RBCs from malaria-affected individuals. These early membrane alterations of uninfected RBCs could be responsible for spreading of infection and RBC lysis during infection.