1. Stable isotope studies have been extremely useful for improving general food web descriptions due to their ability to simultaneously summarize complex trophic networks and track the energy flow through them. However, when considering trophic relationships involving only two or few species, application of general isotopic interpretations based on average fractionation values may easily lead to misleading conclusions. In these cases a more accurate consideration of the current processes involved in the isotopic fractionation should be considered.
2. We investigated the trophic relationships of the crustacean zooplankton assemblage in an alpine lake (Lake Redon, Pyrenees) by means of stable isotopes of carbon and nitrogen and applied information on their life history and biochemical composition in the interpretation.
3. The three species occurring in the lake had distinct isotopic signatures: the two copepod species (the cyclopoid Cyclops abyssorum and the calanoid Diaptomus cyaneus) had higher nitrogen isotopic composition than the cladoceran (Daphnia pulicaria), indicative of a higher trophic position of the two copepods. Most intra-specific isotopic variability was associated with growth, while the effect of metabolic turnover was negligible. The effects of changes in the proportion of lipids was restricted to the adults of the two copepods.
4. Daphnia Juveniles showed ontogenetic shifts in their carbon, and nitrogen isotopic composition. Cyclops copepodites only showed changes in carbon isotopic composition. These isotopic shifts with changes in size were the result of: (i) the prevalence of growth over metabolic turnover as the main factor for isotopic variability and (ii) feeding, during the growth period, on isotopically depleted food in the case of Daphnia, and on isotopically enriched food in the case of Cyclops.
5. The carbon isotopic variation in Cyclops juveniles could be explained by fitting an isotopic growth model that considered that they fed entirely on Daphnia. However this was not the case for nitrogen isotopic variability. Cyclops nitrogen isotopic composition variation and the Cyclops to Daphnia nitrogen isotopic enrichment were closely correlated to the quantity of Daphnia protein and to the dissimilarity in the essential amino acid composition between the two species, which can be interpreted as an indication of consumer nitrogen limitation.