Pyruvate kinase (PK) activity measurements are used to assess the role of carbohydrates in global feeding of mesozooplankton communities inhabiting an estuary plume. As a consequence of a remarkably low freshwater discharge rate, the sea surface layers of the area under estuarine influence showed a very moderate salinity fall and a nearly total depletion in nitrates, whereas higher levels of these nutrients were found in deeper, more saline, layers. Small-scale PK activity variations in mesozooplankton appear to be closely correlated to nitrate integration values within the water column. The results were analysed in comparison with literature reports. The study produced a coherent overall interpretation, which strongly supports the reliability of this new biochemical tool in detecting assimilation of trace carbohydrates in the diet of mesozooplankton.