Housing of individual adult females and feeding regime
Flies were housed in individual cages, which were placed on pupal
collection trays, made from acrylic extrude (described in S2 File), to
allow individual tracking of female reproductive output and survival.
Cameras were fixed below the cages to record weekend activity and to
identify exact larviposition dates. Females were monitored daily between
Monday and Friday and if death occurred, we recorded the date of death.
To determine the exact date of death for females found dead on a Monday
morning, we reviewed video recordings from Friday evening to Monday
morning (S2 File).
Experimental flies were fed following the colony schedule (see above).
To feed individual females, we placed cages on a shallow grid (S2 File).
The grid ensured each female was provided with the same amount of blood
and had equal opportunity for feeding, with c. 100 μl of blood provided
per female. We heated the tray to 37°C and covered the blood with a
silicon membrane, then left flies to feed for 45 minutes, which was
sufficient time for most flies to feed.