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Background and Aims Stable isotope two-source linear mixing models are frequently used to calculate the nutrient-uptake efficiency of carnivorous plants from pooled prey. This study aimed to separate prey into three trophic levels as pooled prey limits statements about the contribution of a specific trophic level to the nutrition of carnivorous plants. Phytoplankton were used as an autotrophic reference for aquatic plants as the lack of suitable reference plants impedes calculation of their efficiency. Methods Terrestrial (Pinguicula) and aquatic (Utricularia) carnivorous plants alongside autotrophic reference plants and potential prey from six sites in Germany and Austria were analysed for their stable isotope natural abundances (δ 15N, δ 13C). A two-source linear mixing model was applied to calculate the nutrient-uptake efficiency of carnivorous plants from pooled prey. Prey preferences were determined using a Bayesian inference isotope mixing model. Key Results Phytophagous prey represented the main contribution to the nutrition of Pinguicula (approx. 55 %), while higher trophic levels contributed a smaller amount (diverse approx. 27 %, zoophagous approx. 17 %). As well as around 48 % nitrogen, a small proportion of carbon (approx. 9 %) from prey was recovered in the tissue of plants. Aquatic Utricularia australis received 29 % and U. minor 21 % nitrogen from zooplankton when applying phytoplankton as the autotrophic reference. Conclusions The separation of prey animals into trophic levels revealed a major nutritional contribution of lower trophic level prey (phytophagous) for temperate Pinguicula species. Naturally, prey of higher trophic levels (diverse, zoophagous) are rarer, resulting in a smaller chance of being captured. Phytoplankton represents an adequate autotrophic reference for aquatic systems to estimate the contribution of zooplankton-derived nitrogen to the tissue of carnivorous plants. The autonomous firing of Utricularia bladders results in the additional capture of phytoplankton, calling for new aquatic references to determine the nutritional importance of phytoplankton for aquatic carnivorous plants. [ABSTRACT FROM AUTHOR]