The Ria Formosa coastal lagoon is a highly productive shallow ecosystem in southern Portugal, subjected to nutrient inputs from anthropogenic and natural sources. Nutrients are major abiotic drivers of phytoplankton in this system, but their effects on phytoplankton assemblages and the occurrence of nutrient limitation are still poorly understood. The main goal of this study was, thus, to evaluate the occurrence, type, and effects of nutrient limitation on phytoplankton community and specific functional groups in the Ria Formosa coastal lagoon. We conducted nutrient enrichment experiments with factorial additions of nitrogen (N) and phosphorus (P) using natural phytoplankton assemblages from distinct locations in the Ria Formosa, throughout a yearly cycle. Phytoplankton composition and abundance were evaluated using inverted and epifluorescence microscopies, and spectrophotometric methods were used for biomass. Limitation was defined as higher phytoplankton growth following enrichment with a particular nutrient in relation to the non-enriched control. The most common type of phytoplankton limitation was simultaneous co-limitation by N and P; diatoms, as r-strategists, were the most frequently limited group. Single N and P limitation, and serial P limitation were also observed, as well as negative responses to nutrient enrichment. Group-specific responses to nutrient enrichment were not reflected in the relative abundance of phytoplankton groups within the whole assemblage, due to the numerical dominance of pico-sized groups (cyanobacteria and eukaryotic picophytoplankton). Ambient nutrient ratios and concentrations did not predict phytoplankton nutrient limitation, given the different nutrient utilisation traits among phytoplankton functional groups. Therefore, nutrient ratios should not be used as indicators of nutrient limitation in eutrophication assessment.
