A nondestructive method for extracting maternally derived egg yolk carotenoids

Maternally deposited carotenoids are a prominent component of egg yolk and are vital for the development and growth of the embryo. In most studies of avian yolk carotenoids, eggs are destructively sampled and this may limit both the number of clutches studied and the research questions addressed. We describe an empirical field trial for a nondestructive biopsy method to extract small samples (0.05 ml) of egg yolk for high-performance liquid chromatography (HPLC) analysis of yolk carotenoid concentrations. We sampled 180 clutches (N= 44 biopsies) of two species of introduced thrushes (genus Turdus) from agricultural habitats in central North Island, New Zealand. Once the protocol was established, all biopsied eggs from clutches that were not depredated or deserted before candling were found to be developing normally after 3-5 d of incubation (N= 28) and all hatched. Biopsy samples (>0.02 g) produced concentrations of yolk carotenoids (and variances) that were statistically indistinguishable from whole yolk destructive samples. In addition, our samples (>0.02 g) confirmed previously reported differences in yolk carotenoid concentrations between the two thrush species and revealed a significant decline in yolk carotenoid concentration with laying order. Further examination of how variability in yolk carotenoid concentration and identity influences offspring sex, success, and survival or, later in life, reproductive success and ability to efficiently incorporate dietary carotenoids into both integument and immune tissues will require larger sample sizes. Studies to date have been restricted by the number of destructive samples that investigators are willing (or permitted) to obtain from wild species. Thus, we hope that our nondestructive method of sampling yolk will promote further examination of the links between carotenoid uptake from the environment and maternal investment in the avian yolk.