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Fig. 2. Iterative HPLC separation (A–E) and behavioural bioassay results
(F–J) of egg foam compounds. Eggs treated with HPLC fractions flanked by
broken red lines give rise to predominately gregariously behaving hatchlings
(corresponding red histograms; median Pgregarious=0.5). In
the coarsest (i.e. primary) separation, all egg foam compounds were parsed
into four fractions (A; F1–F4) and tested in a logistic regression
behavioural bioassay (see text) for gregarizing properties (F). In the
secondary run, components of fraction 3 were collected individually (B) and
tested for bioactivity (G). The tertiary (C,H) and quaternary (D,I) runs were
analogous with still greater specificity. Peak labelling is hierarchical and
reflects peak origin in terms of previous fractions. The top row of histograms
for each trial (F–I) shows the effect of known gregarizing compounds,
sometimes reiterating findings of previous runs, applied to eggs as positive
controls. Histogram bin widths=0.11 Pgregarious units.
Median Pgregarious values are indicated by dotted grey
lines. Peak areas (in black, E), representing F3.2.1.2 (relabelled peak X for
clarity) and close analogues peaks Y and Z, were recorded for later use (see
Fig. 4). Control behavioural
classifications of known crowd- and solitary-reared individuals are provided
(J). Insects from F were randomly drawn from four egg pods, (G) from four egg
pods, (H) from nine egg pods, (I) from 13 egg pods and (J) (solitary-reared)
from four egg pods.
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