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13 result(s) for "Gašparović, Blaženka"
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Bioprospecting of Microalgae Isolated from the Adriatic Sea: Characterization of Biomass, Pigment, Lipid and Fatty Acid Composition, and Antioxidant and Antimicrobial Activity
Marine microalgae and cyanobacteria are sources of diverse bioactive compounds with potential biotechnological applications in food, feed, nutraceutical, pharmaceutical, cosmetic and biofuel industries. In this study, five microalgae, Nitzschia sp. S5, Nanofrustulum shiloi D1, Picochlorum sp. D3, Tetraselmis sp. Z3 and Tetraselmis sp. C6, and the cyanobacterium Euhalothece sp. C1 were isolated from the Adriatic Sea and characterized regarding their growth kinetics, biomass composition and specific products content (fatty acids, pigments, antioxidants, neutral and polar lipids). The strain Picochlorum sp. D3, showing the highest specific growth rate (0.009 h−1), had biomass productivity of 33.98 ± 0.02 mg L−1 day−1. Proteins were the most abundant macromolecule in the biomass (32.83–57.94%, g g−1). Nanofrustulum shiloi D1 contained significant amounts of neutral lipids (68.36%), while the biomass of Picochlorum sp. D3, Tetraselmis sp. Z3, Tetraselmis sp. C6 and Euhalothece sp. C1 was rich in glycolipids and phospholipids (75%). The lipids of all studied microalgae predominantly contained unsaturated fatty acids. Carotenoids were the most abundant pigments with the highest content of lutein and neoxanthin in representatives of Chlorophyta and fucoxanthin in strains belonging to the Bacillariophyta. All microalgal extracts showed antioxidant activity and antimicrobial activity against Gram-negative E. coli and S. typhimurium and Gram-positive S. aureus.
Morpho-physiological adaptations of Leptocylindrus aporus and L. hargravesii to phosphate limitation in the northern Adriatic
The northern Adriatic is highly productive and shallow area characterized by numerous spatio-temporal gradients (e.g. nutrients, salinity, temperature). It is strongly influenced by numerous freshwater inputs, mainly from Po river. Its current systems as well as Po river, generates gradients of phosphate availability with an expressed N/P imbalance and phosphate limitation. A number of recent studies characterized these gradients as major factors affecting abundance and composition of microphytoplankton communities. Focus of this study is on two Leptocylindrus species, Leptocylindrus aporus (F.W. French & Hargraves) D. Nanjappa & A. Zingone 2013 and Leptocylindrus hargravesii D. Nanjappa & A. Zingone 2013. Species belonging to Leptocylindrus genus are frequently observed and have high abundances and also high contributions to the microphytoplankton community in this area. We focused on their morphological and physiological responses to phosphate limitation in situ and also performed in vitro experiments. In this study we report data on species specific growth rates under phosphorus (P) deplete and P rich conditions, localization and characteristics of alkaline phosphate activity, phosphate uptake rates as well as their morphological differences in P deplete versus P rich conditions. Our in vitro experiments showed that both Leptocylindrus species morphologically reacted similarly to phosphorus depletion and showed significantly elongated pervalvar axis in P depleted conditions if compared to P rich conditions. Also average chain lengths increased when in P depleted conditions. Two previously mentioned adaptations indicate their tendency to increase cellular surface areas available for alkaline phosphatase. Chlorophyll fluorescence of both species significantly decreased in P depleted medium. Although both species morphologically reacted similarly, our experiment demonstrated significant differences in physiological reactions to P depleted conditions.
Hyposalinity induces significant polar lipid remodeling in the marine microalga Dunaliella tertiolecta (Chlorophyceae)
Marine microalgae often live in a fluctuating environment including a decrease in salinity caused by global warming induced sea ice melting and freshwater inflows. Under conditions of fluctuating salinity, microalgae have evolved a variety of survival mechanisms such as lipid accumulation and remodeling. The purpose of this study was to investigated the membrane lipid remodeling of the marine green microalga Dunaliella tertiolecta as a short-term acclimation mechanism in response to hyposalinity (20 and 3 PSU) with respect to growth at optimal salinity (38 PSU). We identified 34 lipid species belonging to seven polar lipid classes. Dunaliella tertiolecta accumulates cell lipids and remodels polar lipid classes and their fatty acids composition as response to hypoosmotic stress at 3 PSU. We found that the unsaturation of most polar lipids decreases overall, indicating decreased membrane fluidity and altered permeability, whereas shortening the length of fatty acids of polar lipids is not one of the strategies of D. tertiolecta to cope with the decrease in salinity. Increase in relative content (%) and unsaturation of monogalactosyldiacylglycerols (MGDG) and decrease in relative content (%) and unsaturation of phosphatidylglycerols (PG), suggesting changes in photosynthetic membranes of thylakoids at 20 and 3 PSU. At a very low salinity of 3 PSU, the relative content (%) of phosphatidylinositols (PI) increases, suggesting increased lipid trafficking and signaling in the cells. These changes are statistically significant and we hypothesize that D. tertiolecta is genetically adapted to withstand large salinity fluctuations through polar lipid composition.
Concerted measurements of lipids in seawater and on submicrometer aerosol particles at the Cabo Verde islands: biogenic sources, selective transfer and high enrichments
In the marine environment, measurements of lipids as representative species within different lipid classes have been performed to characterize their oceanic sources and their transfer from the ocean into the atmosphere to marine aerosol particles. The set of lipid classes includes hydrocarbons (HC); fatty acid methyl esters (ME); free fatty acids (FFA); alcohols (ALC); 1,3-diacylglycerols (1,3 DG); 1,2-diacylglycerols (1,2 DG); monoacylglycerols (MG); wax esters (WE); triacylglycerols (TG); and phospholipids (PP) including phosphatidylglycerols (PG), phosphatidylethanolamine (PE), phosphatidylcholines (PC), as well as glycolipids (GL) which cover sulfoquinovosyldiacylglycerols (SQDG), monogalactosyl-diacylglycerols (MGDG), digalactosyldiacylglycerols (DGDG) and sterols (ST). These introduced lipid classes have been analyzed in the dissolved and particulate fraction of seawater, differentiating between underlying water (ULW) and the sea surface microlayer (SML) on the one hand. On the other hand, they have been examined on ambient submicrometer aerosol particle samples (PM1) which were collected at the Cape Verde Atmospheric Observatory (CVAO) by applying concerted measurements. These different lipids are found in all marine compartments but in different compositions. Along the campaign, certain variabilities are observed for the concentration of dissolved (∑DLULW: 39.8–128.5 µg L−1, ∑DLSML: 55.7–121.5 µg L−1) and particulate (∑PLULW: 36.4–93.5 µg L−1, ∑PLSML: 61.0–118.1 µg L−1) lipids in the seawater of the tropical North Atlantic Ocean. Only slight SML enrichments are observed for the lipids with an enrichment factor EFSML of 1.1–1.4 (DL) and 1.0–1.7 (PL). On PM1 aerosol particles, a total lipid concentration between 75.2–219.5 ng m−3 (averaged: 119.9 ng m−3) is measured. As also bacteria – besides phytoplankton sources – influence the lipid concentrations in seawater and on the aerosol particles, the lipid abundance cannot be exclusively explained by the phytoplankton tracer (chlorophyll a). The concentration and enrichment of lipids in the SML are not related to physicochemical properties which describe the surface activity. On the aerosol particles, an EFaer (the enrichment factor on the submicrometer aerosol particles compared to the SML) between 9×104–7×105 is observed. Regarding the individual lipid groups on the aerosol particles, a statistically significant correlation (R2=0.45, p=0.028) was found between EFaer and lipophilicity (expressed by the KOW value), which was not present for the SML. But simple physicochemical descriptors are overall not sufficient to fully explain the transfer of lipids. As our findings show that additional processes such as formation and degradation influence the ocean–atmosphere transfer of both OM in general and of lipids in particular, they have to be considered in OM transfer models. Moreover, our data suggest that the extent of the enrichment of the lipid class constituents on the aerosol particles might be related to the distribution of the lipid within the bubble–air–water interface. The lipids TG and ALC which are preferably arranged within the bubble interface are transferred to the aerosol particles to the highest extent. Finally, the connection between ice nucleation particles (INPs) in seawater, which are already active at higher temperatures (−10 to −15 ∘C), and the lipid classes PE and FFA suggests that lipids formed in the ocean have the potential to contribute to (biogenic) INP activity when transferred into the atmosphere.
Insights into the life strategy of the common marine diatom Chaetoceros peruvianus Brightwell
Chaetoceros peruvianus is a marine diatom species with circumglobal distribution. While frequently observed, it appears never to dominate the marine phytoplankton community hence it can be characterized as a rather opportunistic, generalistic species. Here we present ecological interpretations from a long-term data set on marine microphytoplankton in the northern Adriatic Sea, where the abundancies and relative contributions of C. peruvianus were observed along a set of steep ecological gradients. Limited supply of dissolved inorganic phosphate was identified as the driving ecological factor for this ecosystem. In parallel C. peruvianus was cultivated in monoclonal cultures and its morphological and physiological reaction to replete and phosphorus depleted medium was analysed. C. peruvianus reacted to phosphorus depletion by an increase in cell height and length as well as thickness and length of setae. This morphological reaction included an increase in cellular volume and calculated carbon content. Additionally, it represents the transition between two described morphological varieties, C. peruvianus and C. peruvianus var. robusta. C. peruvianus showed a significant induction of extracellular alkaline phosphatase activity if grown in phosphate depleted medium. Microscopical analysis demonstrated this activity to be located exclusively on the setae of the cells.
Impact of environmental conditions on phospholipid fatty acid composition: implications from two contrasting estuaries
Phospholipid (PL) composition has a tremendous influence on the cell integrity and physiological competency. At the same time, plankton PL make important metabolic fuels for higher trophic levels. The goal of this study was to identify environmental control on PL production and their molecular identity of the suspended particles in two different estuaries. We conducted research in subtropical, eutrophic Wenchang River Estuary in China and temperate pristine, mesotrophic Krka River Estuary in Croatia. In agreement with the more abundant phytoplankton, PL concentrations were much higher in the Wenchang River Estuary (30.3–178.2 μg L−1) than in the Krka River Estuary (8.4–18.8 μg L−1). Given that six PL classes investigated (phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidic acid (PA), phosphatidylinositol (PI), and phosphatidylserine (PS)) have different roles in the cell, we expected their different fatty acid composition in different environments. We found small differences in the fatty acid composition of PC, PG, and PI between two estuaries. These results suggest that the essential fatty acid compositions of these PL in estuarine plankton are relatively constant in order to preserve membrane functions and/or cell processes in which they are involved regardless of environmental conditions. In contrast, PE, PA and PS fatty acid composition substantially differed between two estuaries as well as throughout the salinity gradient in each estuary. This suggests the adaptability of plankton to remodel these PL depending on the environmental conditions and the plankton community structure. Good environmental conditions (favorable N/P ratio, temperature) are important for increased PL content (% in POC and total lipids) in estuarine plankton and increased essential polyunsaturated fatty acid content in PL, which is beneficial to higher trophic levels.
Survival mechanisms of phytoplankton in conditions of stratification-induced deprivation of orthophosphate
Phytoplankton abundance in the northern Adriatic during the summer 2008 indicated that the system was highly productive, in spite of low orthophosphate (PO₄) concentrations. Mechanisms by which phytoplankton adapted to PO₄ deprivation during the summer stratification were studied. In upper, more productive waters, phytoplankton induced high alkaline phosphatase activity (APA) to obtain phosphorus (P) from the dissolved organic pool, and the P turnover time mediated by phytoplankton APA was very short (2 min to 1.5 h). Highaffinity enzymatic activity combined with high hydrolysis rates enabled metabolic flexibility to the phytoplankton in this heterogeneous and fluctuating environment. Another possible mechanism of adaptation to the PO₄ deficit during the summer was a shift toward smaller cells. The smaller nanophytoplankton, supported by higher surface : volume ratios, were presumably able to produce more alkaline phosphatase, an exoenzyme bound to the cell surface. Progressive decrease of large cells and increase of smaller cells in the phytoplankton community during summer supported this hypothesis. In upper waters with low PO₄ concentrations, phytoplankton reduced their P demand by a preferential synthesis of non-phospholipids. In bottom waters, phytoplankton abundance was markedly lower than in upper waters and growth was probably light limited. In these deeper waters with higher PO₄ concentrations, phytoplankton cells did not use APA to obtain P and were able to synthesize more phospholipids. In deeper waters, growth of bigger cells was favored.
The MILAN Campaign
The sea surface microlayer (SML) at the air–sea interface is <1 mm thick, but it is physically, chemically, and biologically distinct from the underlying water and the atmosphere above. Wind-driven turbulence and solar radiation are important drivers of SML physical and biogeochemical properties. Given that the SML is involved in all air–sea exchanges of mass and energy, its response to solar radiation, especially in relation to how it regulates the air–sea exchange of climate-relevant gases and aerosols, is surprisingly poorly characterized. MILAN (Sea Surface Microlayer at Night) was an international, multidisciplinary campaign designed to specifically address this issue. In spring 2017, we deployed diverse sampling platforms (research vessels, radio-controlled catamaran, free-drifting buoy) to study full diel cycles in the coastal North Sea SML and in underlying water, and installed a land-based aerosol sampler. We also carried out concurrent ex situ experiments using several microsensors, a laboratory gas exchange tank, a solar simulator, and a sea spray simulation chamber. In this paper we outline the diversity of approaches employed and some initial results obtained during MILAN. Our observations of diel SML variability show, for example, an influence of (i) changing solar radiation on the quantity and quality of organic material and (ii) diel changes in wind intensity primarily forcing air–sea CO₂ exchange. Thus, MILAN underlines the value and the need of multidiciplinary campaigns for integrating SML complexity into the context of air–sea interaction.
Tracing Nutrients and Organic Matter Changes in Eutrophic Wenchang (China) and Oligotrophic Krka (Croatia) Estuaries: A Comparative Study
Estuaries modify the concentration and composition of riverine nutrients and organic matter (OM), which eventually determine the riverine flux effect to coasts. Nutrients, organic carbon (OC), pigments, and amino acids (AAs) from the samples collected in the eutrophic Wenchang River Estuary (WRE) in China and the oligotrophic Krka River Estuary (KRE) in Croatia were analyzed in order to have a better understanding of how estuaries regulate terrestrial materials. We found a clear increase of dissolved inorganic nitrogen (DIN) concentration and its subsequent decrease due to the removal of DIN (over 100 μM) in the WRE, whereas DIN showed minor variation lengthwise in the KRE, i.e., with the salinity changes, ranging between 1.0 and 5.8 μM. The elevated algae-derived OC, dissolved organic nitrogen, and particulate AAs nitrogen suggest that the OM assimilation may explain approximately one-third of the DIN removal in the WRE, whereas in the oligotrophic KRE, such inorganic to organic transformation is not likely to be significant. Due to the prominent estuarine nitrogen removal/assimilation process, DIN/dissolved inorganic phosphorus (DIP) ratio was as high as 425 in the upper WRE under strong riverine influence, but it declined to as low as 5.4 at the mouth of WRE, that is even lower than DIN/DIP ratio at the mouth of oligotrophic KRE (12). When compared with other rivers worldwide, the Wenchang River showed high nutrients and organic carbon yields. Given the contrasting estuarine process (e.g., DIN removal) between the KRE and the WRE, apparent high nutrient yield from eutrophic rivers should be viewed with caution in ocean studies as the final impact to coastal zone could be similar to the oligotrophic rivers like the KRE.