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.