Microalgae as source of functional ingredients in new-generation foods: challenges, technological effects, biological activity, and regulatory issues.

Microalgae feasibility as food ingredients or source of nutrients and/or bioactive compounds and their health effects have been widely studied. This review aims to provide an overview of the use of microalgae biomass in food products, the technological effects of its incorporation, and their use as a source of health-promoting bioactive compounds. In addition, it presents the regulatory aspects of commercialization and consumption, and the main trends and market challenges Microalgae have stood out as sources of nutritional compounds (polysaccharides, proteins, lipids, vitamins, minerals, and dietary fiber) and biologically active compounds (asthaxanthin, ß-carotene, omega-3 fatty acids). The consumption of microalgae biomass proved to have several health effects, such as hypoglycemic activity, gastroprotective and anti-steatotic properties, improvements in neurobehavioral and cognitive dysfunction, and hypolipidemic properties. Its addition to food products can improve the nutritional value, aroma profile, and technological properties, with important alterations on the syneresis of yogurts, meltability in cheeses, overrun values and melting point in ice creams, physical properties and mechanical characteristics in crisps, and texture, cooking and color characteristics in pastas. However, more studies are needed to prove the health effects in humans, expand the market size, reduce the cost of production, and tighter constraints related to regulations.

The efficiency of microalgae biofilm in the phycoremediation of water from River Kaduna.

This study is aimed at investigating the efficiency of microalgae biofilm in the phycoremediation of water from a polluted river. Freshwater microalgae biofilm inherent in a contaminated petrochemical stream was employed to remediate water from the River Kaduna, which is the largest river in Kaduna town, Kaduna State, Nigeria, and serves as the primary water source in Kaduna town. The results indicate high reduction efficiency of some physicochemical parameters and pollutants (turbidity (71%), conductivity (9.8%), sulfate (37.5%), alkalinity (62.5%), chloride (11.5%), TDS (9.9%), TSS (66.7%), nitrate (42.9%), COD (24%), and BOD (33%), Cd (70.0%), Ni (74.0%) and Pb (71.0%)), indicating the effectiveness of microalgae biofilm in the phycoremediation of water from River Kaduna. According to scanning electron microscope (SEM) observation, the microalgae biofilm has rough surface morphology after the treatment of the river water, which implies that the biofilm was capable of removing the pollutants in water via biosorption. Other characterizations such as XRF, XRD, and FTIR also buttressed that biosorption was the primary removal mechanism of pollutants by microalgae biofilm. Besides, the results also show the production of ROS during the treatment of water from the River Kaduna by the microalgae biofilm. This high concentration of ROS produced during the treatment correlates significantly with pollutant degradation. The GC-MS analysis of the microalgae biofilm shows the involvement of some phytochemicals in the process of pollutant degradation. As a result, microalgae biofilm is a simple and cost-effective method of polluted water phycoremediation with promising applications and future prospects.

Comparing in vivo data and in silico predictions for acute effects assessment of biocidal active substances and metabolites for aquatic organisms.

The purpose of this study was to determine the acute toxicity in aquatic organisms of one biocidal active substance and six metabolites derived from biocidal active substances and to assess the suitability of available QSAR models to predict the obtained values. We have reported the acute toxicity in sewage treatment plant (STP) microorganisms, in the freshwater microalgae Pseudokirchneriella subcapitata and in Daphnia magna following OECD test methods. We have also identified in silico models for acute toxicity of these trophic levels currently available in widely recognized platforms such as VEGA and the OECD QSAR ToolBox. A total of six, four and two models have been selected for Daphnia, algae and microorganisms, respectively. Finally, we have compared the in silico and in vivo data for the seven compounds plus two previously assayed biocidal substances. None of the compounds tested were toxic for Daphnia and STP microorganisms. For microalgae, CGA71019 (1,2,4 triazole) presented an ErC50 value of 38.3 mg/L. The selected in silico models have provided lower EC50 values and are therefore more conservative. Models from the OECD QSAR ToolBox predicted values for 7 out of 9 and for 4 out of 9 chemicals for Daphnia and P. subcapitata, respectively. No predictive models were identified in such platform for STP microorganism's acute effects. In terms of models's specificity, biocide-specific models, developed from curated datasets integrated by biocidal active substances and implemented in VEGA, perform better in the case of microalgae but for Daphnia an alternative, non biocide-specific has revealed a better performance. For STP microorganisms only biocide-specific models have been identified.

High efficiency transformation by electroporation of the freshwater alga Nannochloropsis limnetica.

The microalgal genus of Nannochloropsis is considered one of the most promising organisms for the production of biofuels due to their high lipid content. Transformation systems for marine Nannochloropsis species have been established in the recent decade, however, genetic manipulation of Nannochloropsis limnetica, the only known freshwater species in this genus, is not yet available. Based on established marine Nannochloropsis species electrotransformation protocol, nuclear genetic transformation was established in N. limnetica, meanwhile the appropriate antibiotic selection concentration and electric field strength of electroporation were determined. For the selection of transformants in N. limnetica on plates, 0.07 µg mL-1 of zeocin or 5 µg mL-1 of hygromycin B was proved sufficient, and the transformation efficiency was < 2 × 10-8 with a single pulse ranging from 2200 to 2600 V using 2-mm electroporation cuvettes. Pretreatment of N. limnetica with 10 mM lithium acetate and 3 mM dithiothreitol before electroporation increased transformation efficiency hundreds of times, and the highest transformation efficiency of 10-11 × 10-6 was obtained with an electric field strength of 12,000 V/cm. Our results help to expand the biotechnological applications of this freshwater species and provide means for successful electrotransformation of other microalgae as well. High-efficiency transformation of freshwater Nannochloropsis pretreatment of N. limnetica with 10 mM lithium acetate and 3 mM dithiothreitol before electroporation increased transformation efficiency hundreds of times.

Environmental concentrations of pharmaceuticals directly affect phytoplankton and effects propagate through trophic interactions.

Pharmaceuticals are found in freshwater ecosystems where even low concentrations in the range of ng L-1 may affect aquatic organisms. In the current study, we investigated the effects of chronic exposure to three pharmaceuticals on two microalgae, a potential modulation of the effects by additional inorganic phosphorus (Pi) limitation, and a potential propagation of the pharmaceuticals' effect across a trophic interaction. The latter considers that pharmaceuticals are bioaccumulated by algae, potentially metabolized into more (or less) toxic derivates and consequently consumed by zooplankton. We cultured Acutodesmus obliquus and Nannochloropsis limnetica in Pi-replete and Pi-limited medium contaminated with one of three commonly human used pharmaceuticals: fluoxetine, ibuprofen, and propranolol. Secondly, we tested to what extent first level consumers (Daphnia magna) were affected when fed with pharmaceutical-grown algae. Chronic exposure, covering 30 generations, led to (i) decreased cell numbers of A. obliquus in the presence of fluoxetine (under Pi-replete conditions) (ii) increased carotenoid to chlorophyll ratios in N. limnetica (under Pi-limited conditions), and (iii) increased photosynthetic yields in A. obliquus (in both Pi-conditions). In addition, ibuprofen affected both algae and their consumer: Feeding ibuprofen-contaminated algae to Pi-stressed D. magna improved their survival. We demonstrate, that even very low concentrations of pharmaceuticals present in freshwater ecosystems can significantly affect aquatic organisms when chronically exposed. Our study indicates that pharmaceutical effects can cross trophic levels and travel up the food chain.

Bicarbonate concentration influences carbon utilization rates and biochemical profiles of freshwater and marine microalgae.

Selecting the optimal microalgal strain for carbon capture and biomass production is crucial for ensuring the commercial viability of microalgae-based biorefinery processes. This study aimed to evaluate the impact of varying bicarbonate concentrations on the growth rates, inorganic carbon (IC) utilization, and biochemical composition of three freshwater and two marine microalgal species. Parachlorella kessleri, Vischeria cf. stellata, and Porphyridium purpureum achieved the highest carbon removal efficiency (>85%) and biomass production at 6 g L-1 sodium bicarbonate (NaHCO3), while Phaeodactylum tricornutum showed optimal performance at 1 g L-1 NaHCO3. The growth and carbon removal rate of Scenedesmus quadricauda increased with increasing NaHCO3 concentrations, although its highest carbon removal efficiency (∼70%) was lower than the other species. Varying NaHCO3 levels significantly impacted the biochemical composition of P. kessleri, S. quadricauda, and P. purpureum but did not affect the composition of the remaining species. The fatty acid profiles of the microalgae were dominated by C16 and C18 fatty acids, with P. purpureum and P. tricornutum yielding relatively high polyunsaturated fatty acid content ranging between 14% and 30%. Furthermore, bicarbonate concentration had a species-specific effect on the fatty acid and chlorophyll-a content. This study demonstrates the potential of bicarbonate as an effective IC source for microalgal cultivation, highlighting its ability to select microalgal species for various applications based on their carbon capture efficiency and biochemical composition.

Toxic effects of polystyrene nanoplastics and polycyclic aromatic hydrocarbons (chrysene and fluoranthene) on the growth and physiological characteristics of Chlamydomonas reinhardtii.

While the toxicity of nano-microplastics and polycyclic aromatic hydrocarbons (PAHs) to aquatic organisms is well-studied, their joint impact on microalgae is less explored. This study focused on single and combined effects of PS-NPs (30 nm; concentrations: 2, 5, 10, and 25 mg/L) and two PAHs (chrysene and fluoranthene at 10, 100 µg/L) for 96 h on the accumulation, growth, photosynthetic parameters, and oxidative stress in the Chlamydomonas reinhardtii. The findings revealed that exposure to increasing concentrations of PS-NPs significantly reduced the growth inhibition ratio and chlorophyll-a content after 96 h. Both PAHs (100 µg/L) + PS-NPs (25 mg/L), significantly reduced the growth inhibition ratio and chlorophyll-a levels. Individual and combined exposures of PS-NPs and PAHs can prompt antioxidant responses like SOD, GPx, and GST, as well as an unaffected level of non-enzymatic antioxidant GSH and diminished CAT activity. Furthermore, both PAHs + PS-NPs triggered ROS levels, resulting in cell membrane damage. However, the reduced oxidative effect of LPO of combined exposures can be attributed to the activation of antioxidant defenses. In addition, the microscopic visualization data shows that PS-NPs adhered to the surface of microalgae. Also, PS-NPs reduced the adsorption of PAHs on the surface of C. reinhardtii. Altogether, this study implied that the influence of coexistent PS-NPs should be considered in the environmental risk assessment of PAHs in aquatic environments.

Comparing Pharmacological Potential of Freshwater Microalgae Carotenoids Towards Antioxidant and Anti-proliferative Activity on Liver Cancer (HUH7) Cell Line.

Chemical-based carotenoids have large implications to health as they may cause adverse side effects. Naturally occurring carotenoids mainly from microalgal sources are emerging as excellent substitute to combat cancer diseases. Astaxanthin is the most powerful antioxidant that derived from selected established microalgae with limited yield. Microalgal bioprospecting may provide the high-yielding sources for astaxanthin production. Hence, in the present research, freshwater microalgae Monoraphidium sp. (NCM no. 5585) and Scenedesmus obliquus (NCM no. 5586) were chosen to explore the unique potential of producing astaxanthin. Identification of bioactive metabolites in extracted carotenoid was analyzed through HPLC. Astaxanthin is identified as a major bioactive metabolite in both carotenoid fraction and ß carotene only in Scenedesmus obliquus. Antioxidant potential of microalgal carotenoids was obtained by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and Ferric-reducing antioxidant power (FRAP) assay. The anti-proliferation activity of the extracted carotenoid from Monoraphidium sp. and Scenedesmus obliquus was evaluated against hepatocellular liver carcinoma cell line HUH7 by 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) colorimetric assay. Higher astaxanthin in Monoraphidium sp. leads to boosted antioxidant and anti-proliferation activity contrary to Scenedesmus obliquus that possess both astaxanthin and ß carotene. Though freshwater microalgae have a huge potential to create beneficial metabolites like carotenoids, they are rarely studied in the pharmaceutical industry. This work was the first to investigate the anti-proliferative activity of Monoraphidium sp. and Scenedesmus obliquus carotenoid fraction on the HUH7 hepatocarcinoma cell line.

Metabolic impacts of polystyrene microplastics on the freshwater microalga Microcystis aeruginosa.

Microplastics (plastic particles < 5 mm; MPs) are ubiquitous in aquatic environments but their potential adverse ecological effects on biota remain poorly understood. This is in part because in typical ecotoxicology tests the toxic effects of MPs were found to be limited. To capture the potential find-scale effects of MPs on freshwater organisms, we employed ultra-performance liquid chromatography-tandem mass spectrometry based untargeted metabolomics to investigate the metabolic impact of polystyrene microbeads microplastics (PS-MPs) of different sizes (0.1, 1, 10, 100 µm) and concentrations (1, 10, 100 mg/L) on a common freshwater microalga, Microcystis aeruginosa, after a 96-h exposure test. The phenotype-based results illustrated that while PS-MPs had no discernible effects on microalgal growth and photosynthesis, both oxidative stress and microcystin production were slightly increased. Metabolomics analysis revealed that the PS-MPs altered the global metabolic profile of the microalga. Specially, PS-MPs of larger size and higher concentration induced a larger number of differentially expressed metabolites. The PS-MPs significantly disturbed metabolisms involved in amino acid synthesis, membrane formation, nitrogen storage, and antioxidant defense of the microalga, consistent with the phenotypic observations. These results suggested several perturbed metabolic pathways, especially arginine-related pathways, as the mechanism. Our study showed that the insights provided by metabolomics-based approaches can enhance assessments of the ecological impacts of MPs on freshwater organisms.

Identification and Characterization of a New Microalga Dysmorphococcus globosus-HI from the Himalayan Region as a Potential Source of Natural Astaxanthin.

Synthesized astaxanthin (ASX), stereoisomers of 3S,3'R, 3R,3'R, and 3S,3'S, have over 95% market share and have relatively poor antioxidant and bioactivity properties, with persistent issues in terms of biological functions, health benefits, and biosafety if compared to natural ASX. Bioprospecting of new microalgal strains could be vital for a new source of powerful antioxidant (ASX). In this study, a new algal strain was isolated from the Indian foothills of the Himalayas. Its identity was discerned by morphological and DNA barcode studies. It is a unicellular spheroidal cell-shaped alga with 100-200 µm diameter. The isolate has 93.4% similarity to Dysmorphococcus globosus species based on 18S-rDNA phylogenetic analysis and named as D. globosus-HI (HI stands for Himalayan India). Its growth and major cellular components (carotenoids, carbohydrates, protein, lipids, fatty acid profile, and ASX) were optimized using the seven different culture media. The highest biomass (1.14 g L-1) was observed in the MBBM medium, with a specific growth rate (0.087 day-1), division/day (0.125), and cellular yield (6.16 x 106 cells/mL). The highest carotenoids (1.56 mg g-1), lipids (32.5 mg L-1), and carbohydrates (135.62 mg L-1) were recorded in the 3N-BBM medium. The maximum ω3-FAs (17.78%), ω6-FAs (23.11%), and ω9-FAs (7.06%) were observed in MBBM, JW, and BG-11 medium respectively. The highest amount of antioxidant ASX was accumulated in the 3N-BBM medium (391 mg L-1). It is more than any other known algal species used in the production of natural ASX. The optimized biochemical studies on the D. globosus-HI strain should fulfill the increasing demand for natural ASX for commercial application.

Isolation of Four Microalgal Strains From the Lake Massaciuccoli: Screening of Common Pollutants Tolerance Pattern and Perspectives for Their Use in Biotechnological Applications.

Aquatic ecosystems represent one of the largest reservoirs of phytoplankton accounting for most of the primary production of the Earth. The Lake Massaciuccoli located in Tuscany (Italy) is one of the largest swamps that in ancient times entirely covered the Versilia coastal plain. Despite its peculiar features, especially the eutrophic characteristics, its native microalgal consortia have never been explored up to now. In this work, we isolated and described four autochthonous microalgal strains from different sites in the lake (FB, Idr, CL_Sc, and CL_Ch); the four microalgal strains were identified within the Chlorella sorokiniana clade. We exposed them to ten of the most common or emerging environmental contaminants in order to describe their preliminary response to the tested substances: five metals (As, Fe, Ni, Cu, and Zn), two herbicides (Metolachlor and Sethoxydim), two antibiotics (Ciprofloxacin and Benzylpenicillin) and a non-steroidal anti-inflammatory drug (Ibuprofen). Physiological response of the strains highlighted intraspecific differences; strain CL_Sc was the most tolerant in presence of metals while strain Idr was the most sensitive. All strains were sensitive to sethoxydim and tolerant to metolachlor at all the tested concentrations. Strains FB and Idr were the most sensitive in presence of Ibuprofen while strain CL_Ch was the most sensitive to the highest Benzylpenicillin concentration. Resistance pattern of strain Idr somehow reflects both the phylogenetic and the geographic "isolation" from all other three strains. Finally, optical microscope observation confirmed some differences also in the microalgae morphological aspect. Overall, all the strains showed interesting responses in presence of high concentrations of the tested substances, representing putative interesting candidates for water remediation in wastewater treatment plants.

Isolation and identification of herbivorous ciliates from contaminated microalgal cultures.

Ciliates are a common but understudied group of grazers that can invade microalgal cultures. To estimate the potential impact of ciliates on microalgal culture productivity, the identification of species that can invade these cultures is essential. Furthermore, isolation of these herbivorous ciliates allows to use them in experiments that investigate the impact of ciliate grazing on the productivity of microalgal cultures. The main aims of this study were to isolate and identify ciliates that invade cultures of the freshwater microalgae Chlorella and Chlamydomonas, and to establish a live collection of these ciliates for usage in future experiments. To this end, we optimized a method for isolating ciliates from contaminated microalgal cultures and we developed a new PCR primer set for amplifying the partial 18S rDNA of ciliates belonging to the classes Spirotrichea, Oligohymenophorea and Colpodea. As a result, we isolated 11 ciliates from microalgal enrichment cultures inoculated with non-sterile dust and various freshwater sources. Of these 11 species, 7 were found to be feeding on Chlamydomonas. Ciliate species that fed on Chlorella could not be isolated in this study. Ciliate species feeding on Chlamydomonas were identified based on a combination of morphological observations and molecular analyses of partial 18S rDNA sequences.

Influence of polystyrene microplastics on the growth, photosynthetic efficiency and aggregation of freshwater microalgae Chlamydomonas reinhardtii.

Microplastics are ubiquitous in aquatic ecosystems worldwide, but knowledge on their impacts on phytoplankton, especially freshwater microalgae, is still limited. To investigate this issue, microalgae Chlamydomonas reinhardtii was exposed to polystyrene (PS) microplastics with 4 concentration gradients (5, 25, 50 and 100 mg/L), and the growth, chlorophyll a fluorescence, photosynthetic activities (Fv/Fm), the contents of malondialdehydes (MDA), soluble proteins, extracellular polymeric substances (EPS) and settlement rate were accordingly measured. Results showed that the density of microalgae decreased as the increase of PS microplastics concentrations, and the highest inhibitory rate (IR) was 45.8% on the 7th day under the concentration of 100 mg/L. The high concentration (100 mg/L) of microplastics evidently inhibited the content of EPS released by microalgae into the solution. PS under all dosages tested could reduce both the chlorophyll a fluorescence yields and photosynthetic activities. The scanning electron microscope (SEM) images demonstrated that microplastic beads were wrapped on the surface of microalgae and damaged their membranes, which could suggest the reduction of photosynthetic activities and the increase of soluble proteins and MDA content. The results also showed that PS microplastics could inhibit the settlement of microalgae at the later stage, which also indicated the recovery of microalgae from the toxic environment. Our findings will contribute to understanding the effects of microplastics on freshwater microalgae, as well as evaluating the possible influences of microplastics on aquatic ecosystems.

Long-Term Toxicity of ZnO Nanoparticles on Scenedesmus rubescens Cultivated in Semi-Batch Mode.

The scope of this study was to investigate the toxic effects of zinc oxide (ZnO) nanoparticles (NPs) on freshwater microalgae, in long-term semi-batch feeding mode at two different hydraulic retention times (HRTs) (20 and 40 days). A freshwater microalgae, Scenedesmus rubescens, was employed and exposed to a semi-continuous supply of ZnO NPs at a low concentration of 0.081 mg/L for a period of 28 d. Experiments were conducted under controlled environmental conditions. Τhe impact of ZnO NPs on S. rubescens, which was assessed in terms of nutrient removal, biomass growth, and algal lipid content. Semi-batch mode cultures showed that low ZnO NP concentrations at an HRT of 40 d did not have any negative effect on microalgae growth after the fourth day of culture. In contrast, algal growth was inhibited up to 17.5% at an HRT of 20 d in the presence of ZnO NPs. This might be attributed to the higher flow rate applied and ZnO NPs load. A positive correlation between nutrient removal and microalgae growth was observed. The algal lipid content was, in most cases, higher in the presence of ZnO NPs at both HRTs, indicating that even low ZnO NPs concentration cause stress resulting in higher lipid content.

Characterization of the nano-bio interaction between metallic oxide nanomaterials and freshwater microalgae using flow cytometry.

Since nanomaterials (NMs) are particulate contaminants, their first contact with organisms is a physical encounter ruled by physic-chemical processes that can determinate the potential NMs accumulation, toxicity, and trophic transfer. Freshwater ecosystems often become a final depository for NMs, so they can get in contact with the biota, especially primary organisms as algae. There are almost none comparative studies of this interaction using various NMs in the same conditions. This work identifies, analyzes, and compares the algae-NMs interaction by flow cytometry after a short-term contact test in which three freshwater algae (Raphidocelis subcapitata, Desmodesmus subspicatus, and Chlorella vulgaris) interact individually with a set of twelve metallic oxide NMs. Dose-response profiles and differences in the algae-NMs interaction were found according to each algae species (C. vulgaris had the most affinity, starting the interaction from 0.5 mg/L and D. subspicatus had the less affinity starting at 5 mg/L). Flow cytometry results were confirmed by optical microscopy. Some NMs characteristics were identified as key-factors that govern the algae-NMs interaction: NMs composition (no interaction for SiO2 NMs), surface electric charge (higher interaction for the positively charged NMs and lower interaction for the negatively charged ones) and crystalline form (for TiO2 NMs). The presented method can be useful for a rapid determination of the interaction between free cells organisms as microalgae and (nano)particulate substances.

Adverse physiological and molecular level effects of polystyrene microplastics on freshwater microalgae.

Microplastics have aroused widespread concern because of their adverse effects on aquatic organisms. However, the underlying toxicity mechanisms have not been examined in detail. This study investigated the interactions between polystyrene microplastics (PS-MPs) and the model freshwater microalgae Euglena gracilis. The results of transmission electron microscopy showed that the vacuoles of microalgae were induced after 24 h exposure to 1 mg/L PS-MPs (5 µm and 0.1 µm). Furthermore, PS-MPs significantly (p < 0.05) reduced pigment contents. Moreover, superoxide dismutase activities were significantly (p < 0.05) induced in all PS-MPs treated groups. Peroxidase activities were also significantly (p < 0.05) affected by two sizes of PS-MPs (5 µm and 0.1 µm), indicating that oxidative stress was induced after exposure to PS-MPs. At the molecular level, PS-MPs dysregulated the expression of genes involved in cellular processes, genetic information processing, organismal systems, and metabolisms. The KCS gene and the CTR1 gene may be key pathways to induce adverse effects on the E. gracilis after exposure to 5 µm PS-MPs. These findings will help to elucidate the underlying molecular mechanism of microplastics toxicity on freshwater organisms.

Combined effects of nanoplastics and copper on the freshwater alga Raphidocelis subcapitata.

Nanoplastics are recognized as able to interact with other pollutants including heavy metals, and with natural organic matter, with implications for the potential risks to biota. We investigated the interaction of carboxylated polystyrene nanoparticles (PS-COOH NPs) with copper (Cu) and algal exudates (EPS) and how such interaction could affect Cu toxicity towards the freshwater microalga Raphidocelis subcapitata. PS-COOH NPs behavior in the presence of Cu and EPS was determined by dynamic light scattering (DLS), while PS-COOH NPs surface interaction with Cu ions and EPS was investigated by fluorimetric analysis. ICP-MS was used to test Cu ion adsorption to PS-COOH NPs in the presence and absence of algae. The interaction between PS-COOH NPs and the algal cell wall was assessed by fluorescence microscopy. Short- and long-term toxicity tests were carried out in parallel to assess the impact of PS-COOH NPs on algal growth. Results showed altered nanoparticle surface charge and hydrodynamic diameter following algal EPS exposure, supporting the hypothesis of a protein corona formation. In contrast, no absorption of Cu ions was observed on PS-COOH NPs, either in the presence or absence of algae. No differences on algal growth inhibition were observed between exposure to Cu only, and to Cu in combination with PS-COOH NPs, in short-term as well as long-term tests. However, after 72 h of exposure, the adsorption of PS-COOH NPs to algal cell walls appeared to correspond to morphological alterations, revealing potential disturbances in the mitotic cycle. Our findings confirm the ability of PS-COOH NPs to interact with EPS as shown for other nanomaterials. Environmentally realistic exposure scenarios are thus needed for evaluating nanoplastic toxicity, as nanoparticles will not maintain their pristine nature once released into natural media. Prolonged exposure and use of different end-points such as cell morphological changes and EPS production seem more reliable for the investigation of nanoplastic/algal cell interactions which can drive food chain transfer of nanoplastics and ultimately toxicity.

Algal toxicity of binary mixtures of zinc oxide nanoparticles and tetrabromobisphenol A: Roles of dissolved organic matters.

The present study investigated the impacts of dissolved organic matters (DOM) on joint toxicity involved in zinc oxide nanoparticles (ZnO NPs) and tetrabromobisphenol A (TBBPA) at relevant low-exposure concentrations (<1 mg/L). It was found that ZnO NPs in single and combined systems exhibited severe inhibition effects on a freshwater microalgae Scenedesmus obliquus. However, the presence of DOM slightly alleviated the growth inhibition toxicity induced by the binary mixtures of ZnO NPs and TBBPA. Ultrastructure analysis revealed that ZnO NPs caused structural damage to cells, including plasmolysis, membrane destruction, and the disruption of thylakoid in the chloroplast, regardless of the presence of coexisting substances. Oxidative stress biomarker quantitative analysis and in situ observations indicated that the massive accumulation of reactive oxygen species in the binary mixtures of ZnO NPs and TBBPA caused severe oxidative damage, but the presence of DOM significantly mitigated the damage.

Amino acid composition of algal products and its contribution to RDI.

In this paper, the amino acid profiles of algal products from diverse groups (Cyanophyceae, Chlorophyta, Rhodophyta and Phaeophyta) were established. Contents of essential and non-essential amino acids varied in the range of 22.8-42.3 and 31.0-66.5 (g·16 g(-1)N), respectively. In dependence on daily algal intakes, the highest participation in recommended daily intakes (RDI; related to adult, body weight of 70 kg) of all EAAs was observed in freshwater micro-algal products, especially from Spirulina genus where contribution ranged from 12.6% (Lys, SB) to 38.8% (Thr, S). Generally, Lys was the lowest contributor to RDIs in almost all algal samples except Chlorella pyrenoidosa (C) and Palmaria palmata (D), where Ile and Leu, respectively, were established. Interestingly, the contents of sulfur AAs of both products of the Spirulina genus covered 74.5% (S) and 73.8% (SB) of their RDI. Finally, products from brown seaweeds showed the lowest contributions to the RDIs of all EAAs.

Establishment of a cryopreserved biobank for the Culture Collection of Freshwater Microalgae (CCMA-UFSCar), São Paulo, Brazil / Estabelecimento de um biobanco criopreservado para a Coleção de Culturas de Microalgas de Água-Doce (CCMA-UFSCar), São Paulo, Brasil

Abstract The Culture Collection of Freshwater Microalgae (CCMA-UFSCar, Coleção de Culturas de Microalgas de Água-Doce), based at the Universidade Federal de São Carlos, plays an important role in underpinning Brazilian microalgal research, providing biological materials, substrates and training personnel for a large proportion of the past and current projects in this area. However, recent efforts to expand the culture catalogue/holdings have reached a ceiling because of the logistical practicality of maintaining actively growing cultures. In order to reduce the costs associated with the maintenance regime of cultures, efforts were initiated on the establishment of a cryopreserved biobank for long term maintenance of cultures, thus minimizing the efforts associated with handling of material, as successfully frozen cultures, in theory, could be maintained effectively indefinitely, with the advantage of the stability of cells characteristics. Initial tests were performed on 93 strains including exemplar taxa across the different taxonomic groups in the collection catalogue. The highest levels of success were achieved for the smaller taxa, such as the small green algae; while for the larger and more complex organisms limited to no success was obtained. For the strains tested, over 70% had positive good levels of post-thaw viability and regenerated phenotypically normal cultures.

Resumo A Coleção de Culturas de Microalgas de Água-Doce (CCMA-UFSCar), localizada na Universidade Federal de São Carlos tem um importante papel na sustentação da pesquisa brasileira em microalgas, provendo material biológico, substratos e treinamento de pessoal para uma grande fração dos projetos atuais nessa área. Entretanto, os recentes esforços para expandir o catálogo de culturas atingiram um limite máximo em praticidade logística para a manutenção de culturas metabolicamente ativas. Buscando reduzir os custos do regime de manutenção das culturas, foi-se iniciado o estabelecimento de um banco criopreservado para a manutenção em longo prazo desses organismos, minimizando os gastos com manuseio do material, visto que culturas congeladas com sucesso podem, em teoria, serem mantidas por prazos indeterminados, ainda com a vantagem da estabilidade das características nas culturas. Testes iniciais foram realizados com 93 linhagens, incluindo organismos de diferentes grupos taxonômicos mantidos na coleção de culturas. As maiores taxas de sucesso foram observadas nos organismos menores, como as pequenas algas verdes, enquanto os organismos maiores e mais complexos obtiveram sucesso limitado. Em geral, para as linhagens testadas, acima de 70% dos organismos apresentaram resposta positiva ao processo.