| Summary: | Microbial trophic steps were largely ignored in the estimations of trophic position of consumers. Recent developments in compound-specific stable isotope studies showed that the microbial exchanges may no longer be invisible. Using 24 species of mesopelagic and bathypelagic fishes collected in the North Atlantic, this study applied the analysis of nitrogen isotopes in amino acids to the estimation of their trophic position and of the contribution of microbial and metazoan trophic steps across depth layers. Isotope-based estimates agreed well with diet-based literature values, but the consideration of microbial steps reduced the mismatch between 0.5 and 0.8 trophic positions observed when only the metazoan food web is considered. Microbial trophic steps contributed between 6 to 21% to the overall trophic position of individual species. Body size was positively correlated with trophic position but not with the relative contribution of microbial trophic steps, except in the mesopelagic layer where the microbial contribution decreased with size. These results suggest that current isotope-based estimates of trophic position for marine consumers underestimate true trophic positions because they are based on metazoan-only trophic steps. This research was funded by projects BATHYPELAGIC (CTM2016-78853-R) from the Plan Estatal de I+D+I (Spain), SUMMER (Grant Agreement 817806) and TRIATLAS (Grant Agreement 817578), from the European Union (Horizon 2020 Research and Innovation Programme), and Grant Number IN607A2018/2 from the Axencia Galega de Innovación (GAIN, Xunta de Galicia, Spain).
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