2.3.3. Modifications to Blood Feeding and Oviposition

Only small numbers of F₁ females blood fed initially and repeatedly separating blood fed from non-blood fed females resulted in high mortality. To counter this, all copulated F₁ females, regardless of the strength of interaction with the male, were separated into two 20 × 18 cm plastic cages containing oviposition substrates. The substrates comprised 150 mL clear plastic containers lined with filter paper and a thick layer of cotton wool saturated with distilled water (Figure S3). An extra layer of filter paper was placed above the cotton wool to prevent mosquitoes becoming entangled. Mosquitoes were offered blood daily following the method described in Section 2.2.2, in the hope that some would refeed and survive through multiple gonotrophic cycles. Water and sugar solution were removed no more than two hours before feeding to reduce the risk of desiccation. Oviposition substrates were usually changed every seven days.