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On the day of hatching, nestlings were individually-marked (by clipping a claw) and randomly cross-fostered to either a ��control�� or ��reverse�� experimental brood in the aviary (but never returned to the nest of their genetic parents) (Gilby, Sorato & Griffith, 2012). Each brood consisted of either five or six nestlings, placed into the nest in two groups of two to three nestlings 48 h apart. In ��control�� broods, the first 2�C3 nestlings placed into a nest were from position 1�C3 of the laying sequence, and the latter 2�C3 nestlings from position 4�C6. In ��reverse�� broods this buy BAY 87-2243 pattern was reversed. In total there were 28 successful experimental broods containing 98 nestlings, 15 ��control�� and 13 ��reverse��, which came from 43 different genetic nests. When each nestling was five days old, and again at seven days old, it was removed from its nest and individually placed in an artificial nest on a heat pad to assay begging behaviour. Only a single nestling was taken from a nest at a time in random order, hence the time that each nestling was removed varied (range 11 a.m.�C4 p.m.). Begging was measured in two rounds of three trials each day, with the first round beginning 15 min after the nestling was placed on top of a heat pad in the laboratory. To control for the variation in the level of hunger across different chicks (subject to recent parental allocation) we also measured begging a second time after a standardised feed and rest. Each chick was fed with liquefied egg and biscuit rearing and conditioning food mix until satiation and then left for one hour, before begging behaviour was again assayed. On each meprobamate day the initial begging levels were strongly correlated with the latter ones (Pearson correlation: day 5: r = 0.77, N = 98, P Epacadostat cell line variation in activity levels. During each begging measure, begging was initiated using a standardised procedure with a red pen to tap the nesting material around the nestling, the nestling��s beak and their left and right wing. This routine was conducted three times, with three minutes between each repeat. From video recordings, six different begging variables were quantified: Latency, Duration, Raised head, Head movements, Open mouth and Tongue movements. In total, all of the 98 nestlings (47 males and 51 females) completed the begging assays, producing a total of 568 observations. Further details of the procedures performed on the birds during this relatively early stage of development are described in Gilby, Sorato & Griffith (2012).
