Do mosquitoes transmit the avian malaria-like parasite Haemoproteus? An experimental test of vector competence using mosquito saliva

[Background] The life-cycle of many vector-borne pathogens includes an asexual replication phase in the vertebrate host and sexual reproduction in the insect vector. However, as only a small array of parasites can successfully develop infective phases inside an insect, few insect species are competent vectors for these pathogens. Molecular approaches have identified the potential insect vectors of blood parasites under natural conditions. However, the effectiveness of this methodology for verifying mosquito competence in the transmission of avian malaria parasites and related haemosporidians is still under debate. This is mainly because positive amplifications of parasite DNA in mosquitoes can be obtained not only from sporozoites, the infective phase of the malaria parasites that migrate to salivary glands, but also from different non-infective parasite forms in the body of the vector. Here, we assessed the vectorial capacity of the common mosquito Culex pipiens in the transmission of two parasite genera.[Methods] A total of 1,560 mosquitoes were allowed to feed on five house sparrows Passer domesticus naturally infected by Haemoproteus or co-infected by Haemoproteus/Plasmodium. A saliva sample of the mosquitoes that survived after 13 days post-exposure was taken to determine the presence of parasite DNA by PCR.[Results] Overall, 31.2% mosquito’s head-thorax and 5.8% saliva samples analysed showed positive amplifications for avian malaria parasites. In contrast to Haemoproteus DNA, which was not found in either the body parts or the saliva, Plasmodium DNA was detected in both the head-thorax and the saliva of mosquitoes. Parasites isolated from mosquitoes feeding on the same bird corresponded to the same Plasmodium lineage.[Conclusions] Our experiment provides good evidence for the competence of Cx. pipiens in the transmission of Plasmodium but not of Haemoproteus. Molecular analyses of saliva are an effective method for testing the vector competence of mosquitoes and other insects in the transmission of vector-borne pathogens.This study was funded by projects CGL2012-30759 and CGL2015-65055-P from the Spanish Ministerio de Economia y Competitividad and European Regional Development’s funds (FEDER). We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). R.G.L. was supported by a FPI grant (BES-2013-065274), J.M.P. by a Juan de la Cierva contract, L.G. by a contract under the Excelence Projects from Junta de Andalucía (RNM-7800) and J.Y. by a grant from China Scholarship Council.Peer reviewe