We are currently doing Plasmodium falciparum cultures (NF54 strain) in a perspective to infect Anopheles gambiae by membrane feeding. The Culture evolves up to gametocyte (stage V), but we are struggling to get positive infection by membrane feeding.
We get nice gametocytes in (Stage V) but they seem no infectious for the mosquitoes.
Mosquito infections with P. falciparum are notoriously difficult: only a few labs in the world can reliably do this successfully, so I'd strongly recommend learning your techniques directly from one of these experts! If that's not possible, the detailed methods in Methods In Malaria Research (6th ed. 2013 Kirsten Moll, Akira Kaneko, Arthur Scherf and Mats Wahlgren) might help you. A first step would be to check whether your gametocytes are viable: do they exflagellate efficiently? (For how to check, see papers such as this one: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4249523/)
Thank you very much for your response.
About the gameocyte culture, we observe very rarely the exflagellaation. Does it mean the gametocyte are not viable?
Meanwhile, i'll go through the references you suggest me.
Thank you again
Yes: if the gametocytes are not able to exflagellate (i.e. produce mature male gametes) then there will be no mating with female gametes, no zygotes produced and hence no infection of mosquitoes. Efficient exflagellation is not a guarantee of efficient mosquito infection, but it's a first step! Good luck...
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