A. gambiae mosquitoes were starved for 3–5 h and then fed on NF54 P. falciparum gametocyte cultures (NF54: BEI Resources, MRA-1000; mature gametocytes were adjusted to 0.02% in 40% human red blood cells [O+] containing 60% heat-inactivated human serum [O+]; please refer to the ethics statement) through artificial membranes at 37°C at the Johns Hopkins Malaria Institute Core Facility as previously described (Angleró-Rodríguez et al., 2016 (link); Dong et al., 2018 (link); Trager and Jensen, 1976 (link)). Unfed mosquitoes were removed immediately after blood feeding, and the remaining mosquitoes were incubated at 27°C for 8 d to check oocysts in the midgut, or 14 d to check sporozoites in the SGs.
Protocol detail
Mosquito Infection with P. falciparum
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Corresponding Organization : Howard Hughes Medical Institute
Other organizations : Huazhong Agricultural University
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Variable analysis
independent variables
- Starvation duration (3-5 h)
- Oocyst development in the midgut (8 days)
- Sporozoite development in the salivary glands (14 days)
- NF54 Plasmodium falciparum gametocyte cultures
- Gametocyte concentration (0.02%)
- Red blood cell source (40% human O+ red blood cells)
- Serum source (60% heat-inactivated human O+ serum)
- Feeding method (artificial membranes)
- Feeding temperature (37°C)
- Incubation temperature (27°C)
- None specified
- Unfed mosquitoes removed immediately after blood feeding
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