Many freshwater phytoplankton species have the potential to form transient nuisance blooms that affect water quality and other aquatic biota. Heterotrophic bacteria can influence such blooms via nutrient regeneration but also via antagonism and other biotic interactions. We studied the composition of bacterial communities associated with three bloom-forming freshwater phytoplankton species, the diatom Aulacoseira granulata and the cyanobacteria Microcystis aeruginosa and Cylindrospermopsis raciborskii. Experimental cultures incubated with and without lake bacteria were sampled in three different growth phases and bacterial community composition was assessed by 454-Pyrosequencing of 16S rRNA gene amplicons. Betaproteobacteria were dominant in all cultures inoculated with lake bacteria, but decreased during the experiment. In contrast, Alphaproteobacteria, which made up the second most abundant class of bacteria, increased overall during the course of the experiment. Other bacterial classes responded in contrasting ways to the experimental incubations causing significantly different bacterial communities to develop in response to host phytoplankton species, growth phase and between attached and free-living fractions. Differences in bacterial community composition between cyanobacteria and diatom cultures were greater than between the two cyanobacteria. Despite the significance, major differences between phytoplankton cultures were in the proportion of the OTUs rather than in the absence or presence of specific taxa. Different phytoplankton species favoring different bacterial communities may have important consequences for the fate of organic matter in systems where these bloom forming species occur. The dynamics and development of transient blooms may also be affected as bacterial communities seem to influence phytoplankton species growth in contrasting ways.

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The phylogeny of the family Selenastraceae was investigated by light microscopy, 18S rDNA, rbcL and ITS-2 analyses. Various morphological features traditionally used for species and genera identification were investigated. All selenastracean strains studied have naked pyrenoids within the chloroplast, except the genus Chlorolobion, which presented starch envelope. The molecular analyses showed that no morphological criterion considered so far is significant for the systematics of the Selenastraceae, but a set of features may be suitable to identify the genera Ankistrodesmus and Chlorolobion. Phylogenetic analyses showed the genera Monoraphidium, Kirchneriella and Selenastrum were not monophyletic and not distinguishable as separate genera. The polyphyly of the genus Selenastrum led to the description of two new genera, Curvastrum gen. nov and Messastrum gen. nov.

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Mixotrophy is important to ecosystems functioning. Assuming that limiting resources induce phagotrophy in mixotrophs, we used a factorial experimental design to evaluate how nutrient and light affects phagotrophy in two mixotrophic phytoflagellates belonging to different lineages. We estimated cell-specific grazing rates (CSGR) by analyzing prey ingestion using microscopy and flow cytometry (FC). Furthermore, we tested if the acidotropic probe LysoTracker green (LyTG) can be used to differentiate autotrophs from mixotrophs. Cryptomonas marssonii (cryptophyte) had higher CSGR in high-nutrient treatments. Although it seems counterintuitive, phytoflagellates likely uses phagotrophy to obtain organic growth factors instead of inorganic nutrients when photosynthesis is more favorable. In contrast, CSGR in Ochromonas tuberculata (chrysophyte) increased when light decreased, suggesting that it uses phagotrophy to supplement carbon when autotrophic growth conditions are suboptimal. Measurements of CSGR obtained by FC and microscopy were significantly correlated and displayed the same trend among treatments, although FC rates tended to be higher. Fluorescence with LyTG did not differ from the control in the non-phagotrophic chlorophyte. Contrarily, addition of LyTG significantly increased the fluorescence in chrysophytes and cryptophytes, although no differences were observed among treatments. This approach allowed for differentiation between phagotrophic and non-phagotrophic flagellates but failed to quantify mixotrophy.

, representing 28% of total dissolved neutral carbohydrates (TDnC) on average. Rhamnose concentration was linearly correlated with TDnC concentration and with the abundance of Microcystis aeruginosa, which releases a rhamnose-rich extracellular polysaccharide (EPS). This correlation was slightly stronger when the cell densities of Aulacoseira granulata, Cryptomonas tetrapyrenoidosa and Anabaena spiroides, also rhamnose-rich EPS producers, were included. The enzymatic activity on methylumbelliferyl (MUF)-α-L-rhamnopyranoside (EARh) ranged from 0 to 57.8 nmol MUF l −1 h −1. Glucose concentrations were found to vary from 0 to 4.15 µmol l −1 (27.5% of TDnC), and the enzymatic activity on MUF-β-D-glucopyranoside (EAGl) ranged from 7.4 to 116.9 nmol MUF l −1 h −1. Variation of EARh in time was directly correlated with the density of M. aeruginosa, and with rhamnose concentration in TDnC, while EAGl activity was correlated with density of M. aeruginosa only for variation with depth. Analysis of rhamnose, glucose, TDnC, EARh, EAGl, and bacterial density at each depth individually provided evidence that EPS released was employed as bacterial substrates, despite the permanent presence of TDnC from surface to bottom, owing to its constant production. These results also showed strong evidence that both enzymes were induced by EPS. Our results strongly suggest that the predominant phytoplankton species, known producers of rhamnose and glucose-rich EPS, α-Lrhamnosidase(s) activity and rhamnose concentration in the Barra Bonita Reservoir were coupled.

Amazonian floodplains form complex hydrological networks that play relevant roles in global biogeochemical cycles, and bacterial degradation of the organic matter in these systems is key for regional carbon budget. The Amazon undergoes extreme seasonal variations in water level, which produces changes in landscape and diversifies sources of organic inputs into floodplain systems. Although these changes should affect bacterioplankton community composition (BCC), little is known about which factors drive spatial and temporal patterns of bacterioplankton in these Amazonian floodplains. We used high-throughput sequencing (Illumina MiSeq) of the V3-V4 region of the 16S rRNA gene to investigate spatial and temporal patterns of BCC of two size fractions, and their correlation with environmental variables in an Amazon floodplain lake (Lago Grande do Curuai). We found a high degree of novelty in bacterioplankton, as more than half of operational taxonomic units (OTUs) could not be classified at genus level. Spatial habitat heterogeneity and the flood pulse were the main factors shaping free-living (FL) BCC. The gradient of organic matter from transition zone-lake-Amazon River was the main driver for particle-attached (PA) BCC. The BCC reflected the complexity of the system, with more variation in space than in time, although both factors were important drivers of the BCC in this Amazon floodplain system.