EXPERIMENTS 2-3Evidence for Evolving cue(s) on Ageing Eggs methods A fresh egg mass(0.4-0.5 g) was obtained from thousands of house flies ovipositing on a cotton os (see above) for 3 h. Eggs(0.2 g) were put onto each of two treatment manure oss (see above), and no eggs were put on two control manure oss. one set of treatment and control oss was immediately biotssayedin an oviposition experiment(see General Methods; experiment 2). The other set of treatment and control oss was incubated for 24 h at 24 C and 85% RH and then bioassayed(experiment 3) Results The bioassay flies oviposited significantly more eggs on oss with fresh egg masses than on egg-free control oss(experiment 2, bootstrap test raw mean 81.40%, P< 0.0001; Fig. 2), with 81% of these eggs deposited near fresh egg masses, and only 19% deposited on egg free control oss. However, when the eggs were aged for 24 bioassay flies deposited only 11% of their eggs on oss with aged eggs(and some hatched larvae), which was significantly less than the 89% of eggs deposited on egg-free control oss(experiment 3; bootstrap test: raw mean 10.55%, P<0.0001; Fig. 2). The egg mass' effect on oviposition changed significantly over 24 h(matched pairs bootstrap test: raw mean difference- 70.85%,
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