Anopheles moucheti and Anopheles vinckei Are Candidate Vectors of Ape Plasmodium Parasites, Including Plasmodium praefalciparum in Gabon Christophe Paupy 1,2 *, Boris Makanga 2,3 , Benjamin Ollomo 2 , Nil Rahola 1,2 , Patrick Durand 1 , Julie Magnus 2 , Eric Willaume 4 , Franc ¸ois Renaud 1 , Didier Fontenille 1 , Franck Prugnolle 1,2 * 1 Laboratoire MIVEGEC, UMR 224-5290 CNRS-IRD-UM1-UM2, IRD Montpellier, France, 2 Centre International de Recherches Me ´dicales de Franceville (CIRMF), Franceville, Gabon, 3 Institut de Recherche en Ecologie Tropicale, Libreville, Gabon, 4 Parc de la Le ´ke ´di, ERMAET/COMILOG, Bakoumba, Gabon Abstract During the last four years, knowledge about the diversity of Plasmodium species in African great apes has considerably increased. Several new species were described in chimpanzees and gorillas, and some species that were previously considered as strictly of human interest were found to be infecting African apes. The description in gorillas of P. praefalciparum, the closest relative of P. falciparum which is the main malignant agent of human malaria, definitively changed the way we understand the evolution and origin of P. falciparum. This parasite is now considered to have appeared recently, following a cross-species transfer from gorillas to humans. However, the Plasmodium vector mosquito species that have served as bridge between these two host species remain unknown. In order to identify the vectors that ensure ape Plasmodium transmission and evaluate the risk of transfer of these parasites to humans, we carried out a field study in Gabon to capture Anopheles in areas where wild and semi-wild ape populations live. We collected 1070 Anopheles females belonging to 15 species, among which An. carnevalei, An. moucheti and An. marshallii were the most common species. Using mtDNA-based PCR tools, we discovered that An. moucheti, a major human malaria vector in Central Africa, could also ensure the natural transmission of P. praefalciparum among great apes. We also showed that, together with An. vinckei, An. moucheti was infected with P. vivax-like parasites. An. moucheti constitutes, therefore, a major candidate for the transfer of Plasmodium parasites from apes to humans. Citation: Paupy C, Makanga B, Ollomo B, Rahola N, Durand P, et al. (2013) Anopheles moucheti and Anopheles vinckei Are Candidate Vectors of Ape Plasmodium Parasites, Including Plasmodium praefalciparum in Gabon. PLoS ONE 8(2): e57294. doi:10.1371/journal.pone.0057294 Editor: Ivo Mueller, Walter & Eliza Hall Institute, Australia Received September 19, 2012; Accepted January 20, 2013; Published February 20, 2013 Copyright: ß 2013 Paupy et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was jointly funded by CIRMF (Centre International de Recherches Me ´dicales de Franceville), CNRS (Centre National de la Recherche Scientifique), IRD (Institut de Recherche pour le De ´veloppement) through the program PPR FTH-AC (Programme Pilote Re ´ gional "Changements globaux, Biodiversite ´ et Sante ´ dans les Fore ˆts Tropicales Humides d9 Afrique Centrale"), and by Agence Nationale pour la Recherche (ANR) through the project ORIGIN JCJC-SVSE 7-2012 (Programme Jeunes Chercheurs-Jeunes Chercheuses,Sciences de la Vie, de la Sante ´ et des Ecosyste `mes). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. * E-mail: christophe.paupy@ird.fr (CP); franck.prugnolle@ird.fr (FP) Introduction Recent studies demonstrated that African great apes are infected by different Plasmodium species [1,2], among which four species that are traditionally regarded as human parasites (P. falciparum (3, 4), P. vivax [3,5,6], P. malariae [5,6] and P. ovale [7]. These findings and in particular the discovery in gorillas of parasites that are genetically very close to P. falciparum (these parasites were named P. praefalciparum [2]), have changed paradigms concerning the origin of Plasmodium in humans and emphasized the risk of cross-species exchanges between apes and humans [1,2]. Plasmodium parasites are obligatory mosquito-borne pathogens and Anophelinae mosquitoes might have played and still play a crucial role in their transfer to humans [8]. Inter-human transmission of Plasmodium species is well characterized in sub- Saharan Africa [9]. Conversely, nothing is known about the vector species that ensure Plasmodium transmission in African apes. To identify such vectors, authors [10] analyzed mosquito specimens collected close to nests of wild chimpanzees in Western Uganda, but failed to detect any infected mosquito. Yet, several species, including the Anopheles (An.) moucheti ‘‘sub-species’’ and species from the Anopheles nili group, might be candidate vectors [2]. Indeed, these species are major human malaria vectors, are present in and around forests in West and Central Africa and their distribution range largely overlaps with that of great apes. To identify the vectors of ape Plasmodium and evaluate the risk of transfers to humans, we started a field study in Gabon to capture Anopheles specimens in close proximity to wild and semi-wild ape populations. DNA amplification was used to detect Plasmodium- positive specimens. Results and Discussion Mosquito specimens were collected in the Park of La Le ´ke ´di (Bakoumba, Haut Ogooue ´ Province) and the National Park of La Lope ´ (Ogooue ´ Ivindo Province), in Gabon (Figure 1). Wild chimpanzees (Pan troglodytes troglodytes) and gorillas (Gorilla gorilla gorilla) live in these two wildlife natural reserves and they can be infected by ape Plasmodium (P. gaboni, P. reichenowi in Bakoumba PLOS ONE | www.plosone.org 1 February 2013 | Volume 8 | Issue 2 | e57294