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Evaluating the impacts of climate and fishing on oceanic ecosystems requires an improved understanding of the trophodynamics of pelagic food webs. Our approach was to examine broad-scale spatial relationships among the stable N isotope values of copepods and yellowfin tuna (Thunnus albacares), and to quantify yellowfin tuna trophic status in the food web based on stable-isotope and stomach-contents analyses. Using a generalized additive model fitted to abundance-weighted-average ?¹⁵N values of several omnivorous copepod species, we examined isotopic spatial relationships among yellowfin tuna and copepods. We found a broad-scale, uniform gradient in ?¹⁵N values of copepods increasing from south to north in a region encompassing the eastern Pacific warm pool and parts of several current systems. Over the same region, a similar trend was observed for the ?¹⁵N values in the white muscle of yellowfin tuna caught by the purse-seine fishery, implying limited movement behavior. Assuming the omnivorous copepods represent a proxy for the ?¹⁵N values at the base of the food web, the isotopic difference between these two taxa, “?_YFT-COP,” was interpreted as a trophic-position offset. Yellowfin tuna trophic-position estimates based on their bulk ?¹⁵N values were not significantly different than independent estimates based on stomach contents, but are sensitive to errors in the trophic enrichment factor and the trophic position of copepods. An apparent inshore–offshore, east to west gradient in yellowfin tuna trophic position was corroborated using compound-specific isotope analysis of amino acids conducted on a subset of samples. The gradient was not explained by the distribution of yellowfin tuna of different sizes, by seasonal variability at the base of the food web, or by known ambit distances (i.e. movements). Yellowfin tuna stomach contents did not show a regular inshore–offshore gradient in trophic position during 2003–2005, but the trophic-position estimates based on both methods had similar scales of variability. We conclude that trophic status of yellowfin tuna increased significantly from east to west over the study area based on the spatial pattern of ?_YFT-COP values and the difference between the ?¹⁵N values of glutamic acid and glycine, “trophic” and “source” amino acids, respectively. These results provide improved depictions of trophic links and biomass flows for food-web models, effective tools to evaluate climate and fishing effects on exploited ecosystems.

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