Extraction methods for phycocyanin determination in freshwater filamentous cyanobacteria and their application in a shallow lake

Phycocyanin (PC) is one of the water-soluble accessory pigments of cyanobacteria species, and its concentration in aquatic systems is used to estimate the presence and relative abundance of blue-green algae. PC concentration and the PC/Chl-a ratio of four N2-fixing filamentous cyanobacteria strains (Cylindrospermopsis raciborskii, Anabaena spiroides, Aphanizomenon flos-aquae and Aphanizomenon issatschenkoi) common to Lake Balaton (Hungary) were determined using repeated freezing and thawing. A strong linear correlation was found between the extracted PC and Chl-a concentrations for all strains at high Chl-a concentrations (almost stable PC/Chl-a ratio in the range of 20−100 µg l−1 Chl-a). Extraction of PC and Chl-a from samples with low biomass of cyanobacteria (less than 20 µg l−1 Chl-a) proved to be unreliable using the standard protocol of freeze–thaw cycles (coefficients of variation exceeding 10–15%). In order to find an extraction method that is robust in fresh waters characterized by low algae biomass (e.g. Lake Balaton), the effectiveness of four extraction methods (repeated freeze–thaw method and homogenization with mortar and pestle, Ultrasonic, and Polytron homogenizer) were compared using C. raciborskii. It was found that the efficiency of extraction of phycocyanin was highest when a single freeze–thaw cycle was followed by sonication (25% additional yield compared with using the freeze–thaw method alone). Applying this combined method to surface water samples of Lake Balaton, a strong correlation was found between PC concentration and cyanobacterial biomass (R 2 = 0.9436), whilst the repeated freezing–thawing method found no detectable PC content. Here we show that the combined sonication/freeze–thaw method could be suitable for measuring filamentous cyanobacteria PC content, even at low concentrations; as well as for the estimation of cyanobacterial contribution to total biomass in fresh waters

Comparative algological and bacteriological examinations on biofilms developed on different substrata in a shallow soda lake

According to the European Water Framework Directives, benthic diatoms of lakes are a tool for ecological status assessment. In this study, we followed an integrative sample analysis approach, in order to find an appropriate substratum for the water qualification-oriented biomonitoring of a shallow soda lake, Lake Velencei. Six types of substrata (five artificial and one natural), i.e., andesite, granite, polycarbonate, old reed stems, Plexiglass discs and green reed, were sampled in May and in November. We analysed total alga and diatom composition, chlorophyll a content of the periphyton, surface tension and roughness of the substrata and carbon source utilisation of microbial communities. Water quality index was calculated based on diatom composition. Moreover, using a novel statistical tool, a self-organising map, we related algal composition to substratum types. Biofilms on plastic substrates deviated to a great extent from the stone and reed substrata, with regard to the parameters measured, whereas the biofilms developing on reed and stone substrata were quite similar. We conclude that for water quality monitoring purposes, sampling from green reed during springtime is not recommended, since this is the colonization time of periphyton on the newly growing reed, but it may be appropriate from the second half of the vegetation period. Stone and artificially placed old reed substrata may be appropriate for biomonitoring of shallow soda lakes in both spring and autumn since they showed in both seasons similar results regarding all measured features

Phytoplankton dynamics in relation to seasonal variability and upwelling and relaxation patterns at the mouth of Ria de Aveiro (West Iberian Margin) over a four-year period

From June 2004 to December 2007, samples were weekly collected at a fixed station located at the mouth of Ria de Aveiro (West Iberian Margin). We examined the seasonal and inter-annual fluctuations in composition and community structure of the phytoplankton in relation to the main environmental drivers and assessed the influence of the oceano-graphic regime, namely changes in frequency and intensity of upwelling events, over the dynamics of the phytoplankton assemblage. The samples were consistently handled and a final subset of 136 OTUs (taxa with relative abundance > 0.01%) was subsequently submitted to various multivariate analyses. The phytoplankton assemblage showed significant changes at all temporal scales but with an overriding importance of seasonality over longer-(inter-annual) or shorter-term fluctuations (upwelling-related). Sea-surface temperature, salinity and maximum upwelling index were retrieved as the main driver of seasonal change. Seasonal signal was most evident in the fluctuations of chlorophyll a concentration and in the high turnover from the winter to spring phytoplankton assemblage. The seasonal cycle of production and succession was disturbed by upwelling events known to disrupt thermal stratification and induce changes in the phytoplankton assemblage. Our results indicate that both the frequency and intensity of physical forcing were important drivers of such variability, but the outcome in terms of species composition was highly dependent on the available local pool of species and the timing of those events in relation to the seasonal cycle. We conclude that duration, frequency and intensity of upwelling events, which vary seasonally and inter-annually, are paramount for maintaining long-term phytoplankton diversity likely by allowing unstable coexistence and incorporating species turnover at different scales. Our results contribute to the understanding of the complex mechanisms of coastal phytoplankton dynamics in relation to changing physical forcing which is fundamental to improve predictability of future prospects under climate change.Portuguese Foundation for Science and Technology (FCT, Portugal) [SFRH/BPD/ 94562/2013]; FEDER funds; national funds; CESAM [UID/AMB/50017]; FCT/MEC through national funds; FEDERinfo:eu-repo/semantics/publishedVersio

Phytoplankton chlorophyte structure as related to ENSO events in a saline lowland river (Salado River, Buenos Aires, Argentina)

We analyzed the phytoplankton present in the lower sector of the Salado River (Buenos Aires, Argentina) for 10 years (1995–2005) and detected significant changes occurring in chlorophyte abundance and species richness during La Niña event (1998–1999), which period was analyzed throughout the entire basin (main stream and tributaries). We compared the physicochemical and biologic variables between two El Niño–La Niña–Southern Oscillation (ENSO) periods – El Niño (March 1997–January 1998) and La Niña (May 1998–May 1999) – to identify possible indicators of a relationship between climatic anomalies and chlorophyte performance. Chlorophyte density increased during the La Niña. Under normal or extreme hydrologic conditions, mobile (Chlamydomonas spp.) and nonmobile (Monoraphidium spp.) chlorophytes codominated. These species belonged to Reynolds's functional groups X1 and X2, those typical of nutrient-enriched environments. Comparative analyses between El Niño and La Niña periods indicated significant differences in physicochemical (K+, dissolved polyphenols, particulate reactive phosphorus, alkalinity, pH) and biologic (species diversity and richness, phytoplankton and chlorophyte total densities) variables between the two periods at all basin sites. During the La Niña condition, species richness was greater owing to interconnected shallow lakes and drainage-channel inputs, while the Shannon diversity index was lower because of the high abundance values of Monoraphidium minutum. A detailed analysis of the chlorophytes in the entire basin, indicated that changes in density and species dominance occurred on a regional scale although diverse chlorophyte assemblages were identified in the different sectors of the Salado River basin. After La Niña event, the entire basin had the potential to revert to the previous density values, showing the resilience to global environmental changes and the ability to reestablish the general conditions of stability.Fil: Solari, Lía Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; ArgentinaFil: Gabellone, Nestor Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; ArgentinaFil: Claps, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; ArgentinaFil: Casco, Maria Adela. Universidad Nacional de la Plata. Facultad de Ciencias Naturales y Museo. Division Ficologia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Quaini, Karina Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Limnología "Dr. Raúl A. Ringuelet". Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Instituto de Limnología; ArgentinaFil: Neschuk, Nancy Carolina. Dirección Provincial de Saneamiento y Obras Hidráulicas del Ministerio de Infraestructura, Vivienda y Servicios Públicos de la Provincia de Buenos Aires; Argentin

Dynamics of an intense diatom bloom in the Northern Antarctic Peninsula, February 2016

Diatoms are considered the main base of the Southern Ocean food web as they are responsible for more than 85% of its annual primary production and play a crucial role in the Antarctic trophic structure and in the biogeochemical cycles. Within this context, an intense diatom bloom reaching > 45 mg m−3 of chlorophyll a was registered in the Northern Antarctic Peninsula (NAP) during a late summer study in February 2016. Given that nutrient concentrations and grazing activities were not identified here as limiting factors on the bloom development, the aim of this study was to evaluate the effect of water column structure (stability and upper mixed layer depth) on the phytoplankton biomass and composition in the NAP. The diatom bloom, mainly composed by the large centric Odontella weissflogii (mostly > 70 μm in length), was associated with a local ocean carbon dioxide uptake that reached values greater than −60 mmol m−2 d−1. We hypothesize that the presence of a vertically large water column stability barrier, just below the pycnocline, was the main driver allowing for the development of the intense diatom bloom, particularly in the Gerlache Strait. Contrarily, a shift from diatoms to dinoflagellates (mainly Gymnodiniales < 20 μm) was observed associated with conditions of a highly stable thin layer. The results suggest that a large fraction of this intense diatom bloom is in fast sinking process, associated with low grazing pressure, showing a crucial role of diatoms for the efficiency of the biological carbon pump in this region

Effect of food concentration and type of diet on Acartia survival and naupliar development

We have performed life table experiments to investigate the effects of different food types and concentrations on the larval development and survival up to adulthood of Acartia tonsa. The food species offered comprised a wide taxonomic spectrum: the pigmented flagellates Isochrysis galbana, Emiliania huxleyi, Rhodomonas sp., Prorocentrum minimum, the diatom Thalassiosira weissflogii, grown on medium offering enriched macronutrient concentrations and the ciliate Euplotes sp. initially cultured on Rhodomonas. For the ciliate species, also the functional response was studied. In order to avoid limitation by mineral nutrients, food algae have been taken from the exponential growth phase of the nutrient replete cultures. The suitability of Rhodomonas as a food source throughout the entire life cycle was not a surprise. However, in contrast to much of the recent literature about the inadequacy or even toxicity of diatoms, we found that also Thalassiosira could support Acartia-development through the entire life cycle. On the other hand, Acartia could not complete its life cycle when fed with the other food items, Prorocentrum having adverse effects even when mixed with Rhodomonas and Thalassiosira. Isochrysis well supported naupliar survival and development, but was insufficient to support further development until reproduction. With Emiliania and Euplotes, nauplii died off before most of them could reach the first copepodite stages. Acartia-nauplii showed a behavioral preference for Euplotes-feeding over diatom feeding, but nevertheless Euplotes was an insufficient diet to complete development beyond the naupliar stages

Abundance of a chlorophyll a precursor and the oxidation product hydroxychlorophyll a during seasonal phytoplankton community progression in the Western English Channel

This study presents the first in-situ measurements of the chlorophyll a oxidation product, hydroxychlorophyll a as well as the chlorophyll a precursor, chlorophyll aP276 conducted over an annual cycle. Chlorophyll a oxidation products, such as hydroxychlorophyll a may be associated with the decline of algal populations and can act as an initial step in the degradation of chlorophyll a into products which can be found in the geochemical record, important for studying past climate change events. Here, hydroxychlorophyll a and chlorophyll aP276 were measured at the long-term monitoring station L4, Western Channel Observatory (UK, www.westernchannelobservatory.org) over an annual cycle (2012). Weekly measurements of phytoplankton species composition and abundance enabled detailed analysis of possible sources of hydroxychlorophyll a. Dinoflagellates, 2 diatom species, the prymnesiophyte Phaeocystis spp. and the coccolithophorid Emiliania huxleyi were all associated with hydroxychlorophyll a occurrence. However, during alternate peaks in abundance of the diatoms, no association with hydroxychlorophyll a occurred, indicating that the oxidation of chlorophyll a was dependant not only on species but also on additional factors such as the mode of mortality, growth limiting factor (i.e. nutrient concentration) or phenotypic plasticity. Surface sediment samples contained 10 times more hydroxychlorophyll a (relative to chlorophyll a) than pelagic particulate samples, indicating that more chlorophyll a oxidation occurred during sedimentation or at the sediment-water interface, than in the pelagic environment. In addition, chlorophyll aP276 correlated with chl-a concentration, thus supporting its assignment as a chl-a precursor