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Abstract Methylmercury (MeHg) is known to biomagnify in marine food chains, resulting in higher concentrations in upper trophic level animals than their prey. To better understand how marine copepods, an important intermediate between phytoplankton and forage fish at the bottom of the food chain, assimilate and release MeHg, the authors performed a series of laboratory experiments using the gamma‐emitting radiotracer²⁰³Hg²⁺and Me²⁰³Hg with the calanoid copepodAcartia tonsa. Assimilation efficiencies of Hg²⁺and MeHg ranged from 25% to 31% and 58% to 79%, respectively, depending on algal diets. Assimilation efficiencies were positively related to the fraction of Hg in the cytoplasm of the algal cells that comprised their diet. Efflux rates of Hg²⁺(0.29/d) and MeHg (0.21/d) following aqueous uptake were similar, but efflux rates following dietary uptake were significantly lower for MeHg (0.11–0.22/d) than Hg²⁺(0.47–0.66/d). The calculated trophic transfer factors in copepods were >1 for MeHg and consistently low (≤0.2) for Hg²⁺. The authors used the parameters measured to quantitatively model the relative importance of MeHg sources (water or diet) for copepods and to predict the overall MeHg concentrations in copepods in different marine environments. In general, MeHg uptake from the diet accounted for most of the body burden in copepods (>50%). For an algal diet with a MeHg dry weight bioconcentration factor ≥10⁶, >90% of a copepod's MeHg body burden can be shown to derive from the diet. The model‐predicted MeHg concentrations in the copepods were comparable to independent measurements for copepods in coastal and open‐ocean regions, implying that the measured parameters and model are applicable to natural waters.Environ Toxicol Chem2017;36:1287–1293. © 2016 SETAC