The performance of microwaves irradiation (MAE and VMAE) to extract pigments from two marine microalgae was compared to conventional processes (cold and hot soaking and ultrasound-assisted extraction). Pigments were quantified by RP-HPLC and extraction performance was assessed regarding rapidity, reproducibility and extraction yields. Scanning electron microscopy was used at all extraction steps to assess the impact of the process on microalgal cell integrity. Freeze-drying and pigments extraction preserved microalgae cell integrity (except sonication) and evoked agglutination in superposed cells layers. All processes performed on Dunaliella tertiolecta (chlorophyte) lead to rapid pigments extraction, and equivalent pigments extraction yields, the absence of frustule allowing immediate solvent penetration in microalgae cells. In contrast, presence of the frustule in the diatom Cylindrotheca closterium (bacillariophyte) constituted a mechanical barrier to pigment extraction. MAE was identified as the best extraction process for CC pigments as it combined rapidity, reproducibility, homogeneous heating and high extraction yields.
Engineered nano-sized Cu oxide particles are extensively used in diverse applications. Because aquatic environments are the ultimate "sink" for all contaminants, it is expected that nanoparticles (NP) will follow the same fate. In this study, two marine invertebrates Scrobicularia plana and Hediste diversicolor were chosen as ecotoxicological models. The aim was to evaluate behavioural (burrowing kinetics, feeding rate) and biochemical (biomarkers) responses of S. plana and H. diversicolor exposed in the laboratory to Cu (10 μg L(-1)) added in natural seawater either in the form of engineered nanoparticles (NPs) of CuO or as dissolved Cu in 2% HNO(3). Exposure was characterized by considering (i) the physico-chemical fate of NP (ii) the fraction of labile Cu in experimental media and (iii) Cu bioaccumulation. Results showed high aggregation of CuO NPs in seawater and no additional bioavailable Cu concentrations. Behavioural impairments were observed in S. plana exposed to CuO NPs or soluble Cu whereas in H. diversicolor, only the exposure to soluble Cu led to a burrowing decrease. No obvious neurotoxicity effects were revealed since in both species, no changes in cholinesterasic activity occurred in response to both forms of Cu exposure. Biomarkers of oxidative-stress catalase and glutathione-S-transferase were enhanced in both species whereas superoxide dismutase was increased only in S. plana exposed to CuO NPs. Metallothionein-like protein was increased in bivalves exposed to both forms of Cu. Since, no detectable release of soluble Cu from CuO NPs occurred during the time of experiment, ecotoxicity effects seem to be related to CuO NPs themselves.
Previous studies have demonstrated that bacteria influence microalgal metabolism, suggesting that the selection and characterization of growth-promoting bacteria should offer a new strategy for improving industrial algal cultivation. In the present study, 48 cultivable bacteria were isolated from marine microalgae species and identified using 16S rRNA phylogenetic analysis. The recovered bacteria were found to be members of the α-and γ-Proteobacteria, Cytophaga-Flavobacterium-Bacteroides (CFB) and gram-positive monophyletic clusters. To address the effect of these bacteria on the growth of Dunaliella sp. individually, an experimental high-throughput tool was developed to simultaneously compare replicated associations. A two-step approach was used to monitor growth rate and biomass accumulation of Dunaliella sp. in mixed culture with bacteria, which proved the highthroughput device to be an efficient tool for the selection of growth-promoting bacteria. Depending on the bacterial strain involved, inhibitory effects were recorded for maximal microalgal growth rate, whereas inhibitory and stimulating effects were registered on microalgal biomass accumulation and nitrogen incorporation. Organic nitrogen remineralization by Alteromonas sp. SY007 and Muricauda sp. SY244 is discussed to explain the higher biomass and ammonium incorporation of Dunaliella sp. obtained under nitrogen-limited conditions. These bacteria could be considered as helpers for N accumulation in Dunaliella sp. cells.
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