The transfer of 210Po and 210Pb in the food web of small pelagic fishes (from phytoplankton and zooplankton to anchovy Engraulis encrasicolus and sardine Sardina pilchardus) is investigated in the Gulf of Lion (GoL). We present original data of 210Po and 210Pb activity concentrations, C and N stable isotope ratios, measured (i) from different size classes of phytoplankton and zooplankton during spring and winter in different environments of the GoL, and (ii) in two fish species. Significant spatial patterns based on 210Po, 210Pb activity concentrations and 210Po/210Pb ratios in the different plankton size classes are evidenced by hierarchical clustering, both in spring and winter. This variability, also observed for C and N stable isotopes ratios, is connected to local specific pelagic habitats and hydrodynamics. The sampling strategy suggests that 210Po bioaccumulation in the GoL remains at a constant level from the first (dominated by phytoplankton) to the second trophic level (zooplankton), while 210Pb bioaccumulation shows an increase in winter. Based on stable N isotope ratios and 210Po activity concentrations measured in anchovies and sardines, we evidence 210Po bio-magnification along the trophic food web of these two planktivorous pelagic fishes.