CEPIA brings infrastructures and expertise to collaborators for their research projects. One of the applications is to infect plated hepatic cells with Plasmodium falciparum sporozoites. The procedure to get these sporozoites is the dissect infected mosquitoes, isolate their salivary glands in PBS with penicillin and streptomycin, crush them to liberate the motile sporozoites, count the parasites to determine a specific volume for the inoculum per well. 2 major colonies: Anopheles stephensi and coluzzii, are maintained and depending on which one is used to get sporozoites, bacterial contaminations appear more or less quickly in cell cultures which are incompatible with the in vitro experiments.
Bacteria present in contaminated culture medium has been identified using Maldi-tof and only Elizabethkingia anophelis seems to be present. Other bacteria are probably killed by the large spectrum antibiotics used except E. anophelis which is multiresistant to many of them (Comba IY et al, 2022).
One of our strategy is to produce germ free mosquitoes (Romoli O et al, 2021), to do so, we need to explore the microbiotes of our colonies which could be done using Maldi-tof procedure. As this technic previously needs to cultivate and isolate each type of bacteria, we planned to also used in parallel sequencing to explore more precisely the microbiotes in order to get complete data (Silva BE et al, 2021). Once the list of bacteria will be identified, we will be able to study the effect of a combination of them for a normal development of our mosquitoes, from the eggs to the adult.
