Induction of Cerebral Malaria in Mice

Plasmodium berghei ANKA strain was stored in Alsever's solution in liquid nitrogen until use. A sample was thawed and 100 μL were injected intraperitoneally (i.p.) into a donor mouse of the same age as the experimental group mice. Three days later, blood was collected and 1 × 10⁶ parasitized red blood cells (pRBC) were inoculated i.p. into each animal of the experimental group. Thin blood smears were daily made with a blood drop collected from the tip of the tail, stained according to the Panoptic method (Laborclin, Brazil) and examined under a light microscope (BH2, Olympus: Melville, New York, USA) with an oil immersion lens (1,000× final magnification). Parasitaemia levels were measured by counting the percentage of pRBC in at least 2,000 RBC. Mice were observed three times a day and CM diagnosis was made based on presentation of neurological clinical signs (ataxia, disorientation, paraplegia, roll-over and coma) on days 5 - 12 post-infection.