Shifts in isotopic signatures of animals with complex life-cycles can complicate conclusions on cross-boundary trophic links

Stable isotopes (¹³C and ¹⁵N) are widely applied in studies of trophic links. We used this method to investigate the contribution of aquatic and terrestrial prey to the diet of riparian predatory arthropods in two mountain headwater streams in Colorado, USA. Aquatic and terrestrial prey and riparian predators were collected during summer 2009. To evaluate the reliability of conclusions based on stable isotope ratios, we compared the isotopic signatures of aquatic larval and terrestrial adult stages of three abundant stream insect species and assessed variation in mixing model estimates for spider diet composition under varying assumptions for trophic fractionation. Adult isotopic signatures of some aquatic prey species were indistinguishable from those of prey species with exclusively terrestrial life histories (stoneflies: ¹³C and ¹⁵N, chironomids: ¹³C). Other prey had distinctly aquatic isotopic signatures as both larvae and adults (a mayfly and a caddisfly). There was no evidence that prey with aquatic isotopic signatures contributed to the diet of the spiders near one stream. For the other stream, mixing model analysis suggested that chironomids were included in the diets of lycosid, linyphiid and liocranid spiders. Reliable estimates of the contributions of prey sources were compromised by the sensitivity of mixing models to assumptions on trophic fractionation and the presence of “isotopically cryptic” prey. This study emphasizes the importance of supporting isotope-based studies on cross-boundary trophic links with data on isotopic shifts in prey with complex life cycles and assessment of fractionation rates specific to the study system.