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Phytoplankton are dominant primary producers and key indicators in aquatic ecosystems. Understanding the controlling factors on the structure of phytoplankton assemblages is fundamental, but particularly challenging at the land–ocean interface. To identify the patterns and predictors of phytoplankton assemblage structure in the Ria Formosa coastal lagoon (south Portugal), this study combined phytoplankton abundance along a transect between the discharge point of a wastewater treatment plant and a lagoon inlet, over two years, with physico-chemical, hydrographic, and meteo-oceanographic variables. Our study identified 147 operational taxonomic units (OTUs), and planktonic diatoms (60–74%) and cryptophyceans (17–25%) dominated the phytoplankton in terms of abundance. Despite strong lagoon hydrodynamics, and the lack of spatial differences in the phytoplankton abundance and most diversity metrics, the multivariate analysis revealed differences in the assemblage structure between stations (p < 0.001) and seasons (p < 0.01). Indicator analysis identified cryptophyceans as lagoon generalists, and 11 station-specific specialist OTUs, including Kryptoperidinium foliaceum and Oscillatoriales (innermost stations) and potentially toxigenic species (Pseudo-nitzschia and Dinophysis; outer lagoon station, p < 0.05). Water temperature, pH, and nutrients emerged as the variables that best explained the changes in the phytoplankton assemblage structure (p < 0.001). Our findings provide insight into the relevance of local anthropogenic and natural forcings on the phytoplankton assemblage structure and can be used to support the management of RF and other coastal lagoons. [ABSTRACT FROM AUTHOR]

Phytoplankton play a key role in marine ecosystems, making the accurate quantification of functional groups/species, using standardized microscopy techniques, essential in many research efforts. In this study we aimed to evaluate the effects of (a) storage time on the abundance of microphytoplankton in samples fixed with acid Lugol's solution; (b) storage time on the abundance of pico- and nanophytoplankton in frozen microscopy slides; (c) storage time on the abundance of pico- and nanophytoplankton in fixed refrigerated samples and (d) storage protocol (refrigerated versus frozen) on the abundance of pico- and nanophytoplankton. Microphytoplankton were analyzed using inverted microscopy, and pico- and nanophytoplankton using epifluorescence microscopy. Results indicate storage time negatively impacted the abundances of all phytoplankton size classes; however, effects were group-specific. For accurate abundance estimates, we suggest that samples fixed with acid Lugol's should be analyzed within 30 and 180 days, for dinoflagellates/total microphytoplankton and diatoms, respectively. For picoeukaryotes, glutaraldehyde fixed samples should be kept refrigerated for up to 4 days, and slides should be prepared immediately before observation. It is recommended that authors specify the exact lag times between sample collection, fixation and analysis, to allow the comparability of phytoplankton datasets across different studies and/or monitoring programs. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Changes in temperature and CO2 are typically associated with climate change, but they also act on shorter time scales, leading to alterations in phytoplankton physiology and community structure. Interactions among stressors may cause synergistic or antagonistic effects on phytoplankton dynamics. Therefore, the main goal of this work is to understand the short-term isolated and interactive effects of warming and high CO2 on phytoplankton nutrient consumption, growth, production, and community structure in the Ria Formosa coastal lagoon (southern Portugal). We performed microcosm experiments with temperature and CO2 manipulation, and dilution experiments under temperature increase, using winter phytoplankton assemblages. Phytoplankton responses were evaluated using inverted and epifluorescence microscopy. Overall, phytoplankton growth and microzooplankton grazing on phytoplankton decreased with warming. Negative antagonist interactions with CO2 alleviated the negative effect of temperature on phytoplankton and cryptophytes. In contrast, higher temperature benefited smaller-sized phytoplankton, namely cyanobacteria and eukaryotic picophytoplankton. Diatom growth was not affected by temperature, probably due to nutrient limitation, but high CO2 had a positive effect on diatoms, alleviating the effect of nutrient limitation. Results suggest that this winter phytoplankton assemblage is well acclimated to ambient conditions, and short-term increases in temperature are detrimental, but can be alleviated by high CO2. [ABSTRACT FROM AUTHOR]