Wild-living chimpanzees and gorillas harbor a multitude of
Plasmodium
species, including six of the subgenus
Laverania
, one of which served as the progenitor of
Plasmodium falciparum
. Despite the magnitude of this reservoir, it is unknown whether apes represent a source of human infections. Here, we used
Plasmodium
species-specific PCR, single-genome amplification, and 454 sequencing to screen humans from remote areas of southern Cameroon for ape
Laverania
infections. Among 1,402 blood samples, we found 1,000 to be
Plasmodium
mitochondrial DNA (mtDNA) positive, all of which contained human parasites as determined by sequencing and/or restriction enzyme digestion. To exclude low-abundance infections, we subjected 514 of these samples to 454 sequencing, targeting a region of the mtDNA genome that distinguishes ape from human
Laverania
species. Using algorithms specifically developed to differentiate rare
Plasmodium
variants from 454-sequencing error, we identified single and mixed-species infections with
P. falciparum
,
Plasmodium malariae
, and/or
Plasmodium ovale
. However, none of the human samples contained ape
Laverania
parasites, including the gorilla precursor of
P. falciparum
. To characterize further the diversity of
P. falciparum
in Cameroon, we used single-genome amplification to amplify 3.4-kb mtDNA fragments from 229 infected humans. Phylogenetic analysis identified 62 new variants, all of which clustered with extant
P. falciparum
, providing further evidence that
P. falciparum
emerged following a single gorilla-to-human transmission. Thus, unlike
Plasmodium knowlesi
-infected macaques in southeast Asia, African apes harboring
Laverania
parasites do not seem to serve as a recurrent source of human malaria, a finding of import to ongoing control and eradication measures.