The aim of this study was to quantify the N removal efficiency of an Ulva-based phytotreatment system receiving wastewaters from a land-based fish farm (Orbetello, Italy), to identify the main biogeochemical pathways involved and to provide basic guidelines for treatment implementation and management. Fluxes of O2 and nutrients in bare and in Ulva colonised sediments were assessed by light/dark core incubations; denitrification by the isotope pairing technique and Ulva growth by in situ incubation of macroalgal disks in cages. O2 and nutrient budgets were estimated as sum of individual processes and further verified by 24-h investigations of overall inlet and outlet loads. Ulva uptake (up to 7.8 mmol N m−2 h−1) represented a net sink for water column and regenerated View the MathML source whilst N removal via denitrification (10–170 μmol N m−2 h−1) accounted for a small percentage of inorganic nitrogen load (