Five Aotus monkeys and two chimpanzees were infected with Plasmodium malariae isolated from a patient who acquired her infection approximately 50 years ago. All animals were splenectomized. The chimpanzees supported the highest parasite densities of 22,271/microliters and 18,544/microliters. Three Aotus monkeys with a previous history of infection with P. vivax had maximum parasite counts of from 1,818/microliters to 2,909/microliters, whereas two monkeys not previously infected had maximum parasite counts of 6,908/microliters. The establishment of new isolates in these animals aides the development of diagnostic probes and the identification of areas of antigenic variation within the species.