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Background  Direct membrane feeding assays (DMFA) are an important tool to study parasite transmission to mosquitoes. Mosquito feeding rates in these artificial systems require optimization, as there are a number of factors that potentially influence the feeding rates and there are no standardized methods that apply to all anopheline species.    Methods  A range of parameters prior to and during direct membrane feeding (DMF) were evaluated for their impact on  Anopheles farauti  sensu stricto feeding rates, including the starving conditions and duration of starving prior to feeding, membrane type, DMF exposure time, mosquito age, feeding in the light versus the dark, blood volume, mosquito density and temperature of water bath.    Results  The average successful DMFA feeding rate for  An. farauti  s.s. colony mosquitoes increased from 50 to 85% when assay parameters were varied. Overnight starvation and Baudruche membrane yielded the highest feeding rates but rates were also affected by blood volume in the feeder and the mosquito density in the feeding cups. Availability of water during the pre-feed starvation period did not significantly impact feeding rates, nor did the exposure duration to blood in membrane feeders, the age of mosquitoes (3, 5 and 7 days post-emergence), feeding in the light versus the dark, or the temperature (34 °C, 38 °C, 42 °C and 46 °C) of the water bath.    Conclusion  Optimal feeding conditions in  An. farauti  s.s. DMFA were to offer 50 female mosquitoes in a cup (with a total surface area of ~ 340 cm 2  with 1 mosquito/6.8 cm 2 ) that were starved overnight 350–500 µL of blood (collected in heparin-coated Vacutainer tubes) per feeder in feeders with a surface area ~ 5 cm 2  (with a maximum capacity of 1.5 mL of blood) via a Baudruche membrane, for at least 10–20 min.    Graphical AbstractSupplementary Information  The online version contains supplementary material available at 10.1186/s13071-021-04842-y.

Optimization of the feeding rate of Anopheles farauti s.s. colony mosquitoes in direct membrane feeding assays