A Comprehensive Review of Microalgae and Cyanobacteria-Based Biostimulants for Agriculture Uses.

Significant progress has been achieved in the use of biostimulants in sustainable agricultural practices. These new products can improve plant growth, nutrient uptake, crop yield and quality, stress adaptation and soil fertility, while reducing agriculture's environmental footprint. Although it is an emerging market, the biostimulant sector is very promising, hence the increasing attention of the scientific community and agro-industry stakeholders in finding new sources of plant biostimulants. Recently, pro- and eucaryotic microalgae have gained prominence and can be exploited as biostimulants due to their ability to produce high-value-added metabolites. Several works revealed the potential of microalgae- and cyanobacteria-based biostimulants (MCBs) as plant growth promoters and stress alleviators, as well as encouraging results pointing out that their use can address current and future agricultural challenges. In contrast to macroalgae biostimulants, the targeted applications of MBs in agriculture are still in their earlier stages and their commercial implementation is constrained by the lack of research and cost of production. The purpose of this paper is to provide a comprehensive overview on the use of this promising new category of plant biostimulants in agriculture and to highlight the current knowledge on their application prospects. Based on the prevailing state of the art, we aimed to roadmap MCB formulations from microalgae and cyanobacteria strain selection, algal biomass production, extraction techniques and application type to product commercialization and farmer and consumer acceptance. Moreover, we provide examples of successful trials demonstrating the beneficial applications of microalgal biostimulants as well as point out bottlenecks and constraints regarding their successful commercialization and input in sustainable agricultural practices.

Chlorella pyrenoidosa as a potential bioremediator: Its tolerance and molecular responses to cadmium and lead.

Heavy metal contamination negatively affects plants and animals in water as well as soils. Some microalgae can remove heavy metal contaminants from wastewater. The aim of this study was to screen green microalgae (GM) to identify those that tolerate high concentrations of toxic heavy metals in water as possible candidates for phytoremediation. Analyses of the tolerance, physiological parameters, ultrastructure, and transcriptomes of GM under Cd/Pb treatments were conducted. Compared with the other GM, Chlorella pyrenoidosa showed stronger tolerance to high concentrations of Cd/Pb. The reduced glutathione content and peroxidase activity were higher in C. pyrenoidosa than those in the other GM. Ultrastructural observations showed that, compared with other GM, C. pyrenoidosa had less damage to the cell surface and interior under Cd/Pb toxicity. Transcriptome analyses indicated that the "peroxisome" and "sulfur metabolism" pathways were enriched with differentially expressed genes under Cd/Pb treatments, and that CpSAT, CpSBP, CpKAT2, Cp2HPCL, CpACOX, CpACOX2, and CpACOX4, all of which encode antioxidant enzymes, were up-regulated under Cd/Pb treatments. These results show that C. pyrenoidosa has potential applications in the remediation of polluted water, and indicate that antioxidant enzymes contribute to Cd/Pb detoxification. These findings will be useful for producing algal strains for the purpose of bioremediation in water contamination.

Impact of green and blue-green light on the growth, pigment concentration, and fatty acid unsaturation in the microalga Monoraphidium braunii.

The spectral composition of light is an important factor for the metabolism of photosynthetic organisms. Several blue light-regulated metabolic processes have already been identified in the industrially relevant microalga Monoraphidium braunii. However, little is known about the spectral impact on this species' growth, fatty acid (FA), and pigment composition. In this study, M. braunii was cultivated under different light spectra (white light: 400-700 nm, blue light: 400-550 nm, green light: 450-600 nm, and red light: 580-700 nm) at 25°C for 96 h. The growth was monitored daily. Additionally, the FA composition, and pigment concentration was analyzed after 96 h. The highest biomass production was observed upon white light and red light irradiation. However, green light also led to comparably high biomass production, fueling the scientific debate about the contribution of weakly absorbed light wavelengths to microalgal biomass production. All light spectra (white, blue, and green) that comprised blue-green light (450-550 nm) led to a higher degree of FA unsaturation and a greater concentration of all identified pigments than red light. These results further contribute to the growing understanding that blue-green light is an essential trigger for maximized pigment concentration and FA unsaturation in green microalgae.

Chlamydomonas agloeformis from the Ecuadorian Highlands: Nutrients and Bioactive Compounds Profiling and In Vitro Antioxidant Activity.

Green microalgae are single-celled eukaryotic organisms that, in recent years, are becoming increasingly important in the nutraceutical, cosmetic, and pharmaceutical fields because of their high content of bioactive compounds. In this study, a particular green microalga was isolated from freshwater highland lakes of Ecuador and morphologically and molecularly identified as Chlamydomonas agloeformis (ChA), and it was studied for nutritional and nutraceutical properties. The phenolic composition and the fatty acids profile of lyophilized cells were determined. The methanolic extract was analyzed for the phenolic compounds profile and the antioxidant capacity by means of in vitro tests. Finally, Human Microvascular Endothelial Cells (HMEC-1) were exploited to explore the capacity of ChA to reduce the endothelial damage induced by oxidized LDL-mediated oxidative stress. The extract showed a good antioxidant ability thanks to the high content in polyphenolic compounds. The observed decrease in HMEC-1 cells endothelial damage also was probably due to the antioxidant compounds present in the extract. Based on the outcomes of our in vitro assays, ChA demonstrated to be a promising source of bioactive compounds possessing exceptional antioxidant capacities which make it a prospective functional food.

Study of the Antimicrobial Potential of the Arthrospira platensis, Planktothrix agardhii, Leptolyngbya cf. ectocarpi, Roholtiella mixta nov., Tetraselmis viridis, and Nanofrustulum shiloi against Gram-Positive, Gram-Negative Bacteria, and Mycobacteria.

The incidence of diseases brought on by resistant strains of micro-organisms, including tuberculosis, is rising globally as a result of the rapid rise in pathogenic micro-organism resistance to antimicrobial treatments. Secondary metabolites with potential for antibacterial activity are produced by cyanobacteria and microalgae. In this study, gram-positive (S. aureus, E. faecalis) and gram-negative (K. pneumoniae, A. baumannii, P. aeruginosa) bacteria were isolated from pulmonary tuberculosis patients receiving long-term antituberculosis therapy. The antimicrobial potential of extracts from the cyanobacteria Leptolyngbya cf. ectocarpi, Planktothrix agardhii, Arthrospira platensis, Rohotiella mixta sp. nov., Nanofrustulum shiloi, and Tetraselmis (Platymonas) viridis Rouchijajnen was evaluated. On mouse splenocytes and peritoneal macrophages, extracts of cyanobacteria and microalgae had inhibitory effects. In vitro studies have shown that cyanobacteria and microalgae extracts suppress the growth of bacteria and mycobacteria. At the same time, it has been demonstrated that cyanobacterial and microalgal extracts can encourage bacterial growth in a test tube. Additionally, the enhanced fucoxanthin fraction significantly reduced the development of bacteria in vitro. In a mouse experiment to simulate tuberculosis, the mycobacterial load in internal organs was considerably decreased by fucoxanthin. According to the information gathered, cyanobacteria and microalgae are potential sources of antibacterial compounds that can be used in the manufacturing of pharmaceutical raw materials.

Bioactive Compounds with Pesticide Activities Derived from Aged Cultures of Green Microalgae.

The excessive use of synthetic pesticides has caused environmental problems and human health risks and increased the development of resistance in several organisms. Allelochemicals, secondary metabolites produced as part of the defense mechanisms in plants and microorganisms, are an attractive alternative to replace synthetic pesticides to remediate these problems. Microalgae are natural producers of a wide range of allelochemicals. Thus, they provide new opportunities to identify secondary metabolites with pesticide activities and an alternative approach to discover new modes of action and circumvent resistance. We screened 10 green microalgae strains belonging to the Chlorophyta phylum for their potential to inhibit the growth of photosynthetic and nonphotosynthetic organisms. Bioassays were established to assess microalgae extracts' effectiveness in controlling the growth of Chlorella sorokiniana, Arabidopsis thaliana, Amaranthus palmeri, and the model nematode Caenorhabditis elegans. All tested strains exhibited herbicidal, nematocidal, or algicidal activities. Importantly, methanol extracts of a Chlamydomonas strain effectively controlled the germination and growth of a glyphosate-resistant A. palmeri biotype. Likewise, some microalgae extracts effectively killed C. elegans L1 larvae. Comprehensive metabolic profiling using LC-MS of extracts with pesticide activities showed that the metabolite composition of Chlamydomonas, Chlorella, and Chloroidium extracts is diverse. Molecules such as fatty acids, isoquinoline alkaloids, aldehydes, and cinnamic acids were more abundant, suggesting their participation in the pesticide activities.

Current Nuclear Engineering Strategies in the Green Microalga Chlamydomonas reinhardtii.

The green model microalga Chlamydomonas reinhardtii recently emerged as a sustainable production chassis for the efficient biosynthesis of recombinant proteins and high-value metabolites. Its capacity for scalable, rapid and light-driven growth in minimal salt solutions, its simplicity for genetic manipulation and its "Generally Recognized As Safe" (GRAS) status are key features for its application in industrial biotechnology. Although nuclear transformation has typically resulted in limited transgene expression levels, recent developments now allow the design of powerful and innovative bioproduction concepts. In this review, we summarize the main obstacles to genetic engineering in C. reinhardtii and describe all essential aspects in sequence adaption and vector design to enable sufficient transgene expression from the nuclear genome. Several biotechnological examples of successful engineering serve as blueprints for the future establishment of C. reinhardtii as a green cell factory.

In Vitro Characterization of Antioxidant, Antibacterial and Antimutagenic Activities of the Green Microalga Ettlia pseudoalveolaris.

Recently, green microalgae have gained importance due to their nutritional and bioactive compounds, which makes them some of the most promising and innovative functional foods. The aim of this study was to evaluate the chemical profile and the in vitro antioxidant, antimicrobial and antimutagenic activity of an aqueous extract of the green microalga Ettlia pseudoalveolaris, obtained from the freshwater lakes of the Ecuadorian Highlands. Human microvascular endothelial cells (HMEC-1) were used to determine the ability of the microalga to reduce the endothelial damage caused by hydrogen peroxide-induced oxidative stress. Furthermore, the eukaryotic system Saccharomyces cerevisiae was used to evaluate the possible cytotoxic, mutagenic and antimutagenic effect of E. pseudoalveolaris. The extract showed a notable antioxidant capacity and a moderate antibacterial activity mostly due to the high content in polyphenolic compounds. It is likely that the antioxidant compounds present in the extract were also responsible for the observed reduction in endothelial damage of HMEC-1 cells. An antimutagenic effect through a direct antioxidant mechanism was also found. Based on the results of in vitro assays, E. pseudoalveolaris proved to be a good source of bioactive compounds and antioxidant, antibacterial and antimutagenic capacities making it a potential functional food.

Structural properties of the extracellular biopolymer (ß-D-xylo-α-D-mannan) produced by the green microalga Gloeocystis vesiculosa Nägeli.

Many species of microalgae produce a relatively diverse range of metabolites that are interesting for biotechnological applications, and among them exopolysaccharides attract attention due to their structural complexity, biological activities, biodegradability or biocompatibility. An exopolysaccharide of high molecular weight (Mp) of 6.8 × 105 g/mol was obtained by cultivation of the freshwater green coccal microalga Gloeocystis vesiculosa Nägeli 1849 (Chlorophyta). Chemical analyses revealed a dominance of Manp (63.4 wt%), Xylp and its 3-O-Me-derivative (22.4 wt%), and Glcp (11.5 wt%) residues. The results of the chemical and NMR analyses showed an alternating branched 1,2- and 1,3-linked α-D-Manp backbone terminated by a single ß-D-Xylp and its 3-O-methyl derivative at O2 of the 1,3-linked α-D-Manp residues. The α-D-Glcp residues were found mainly as 1,4-linked and to a lesser extent as the terminal sugar, indicating partial contamination of ß-D-xylo-α-D-mannan with amylose (∼10 wt%) in G. vesiculosa exopolysaccharide.

Growth Parameters of Various Green Microalgae Species in Effluent from Biogas Reactors: The Importance of Effluent Concentration.

The use of liquid waste as a feedstock for cultivation of microalgae can reduce water and nutrient costs and can also be used to treat wastewater with simultaneous production of biomass and valuable products. This study applied strategies to treat diluted anaerobic digester effluent (ADE) as a residue of biogas reactors with moderate (87 ± 0.6 mg L-1; 10% ADE) and elevated NH4+-N levels (175 ± 1.1 mg L-1; 20% ADE). The effect of ADE dilution on the acclimatization of various microalgae was studied based on the analysis of the growth and productivity of the tested green algae. Two species of the genus Chlorella showed robust growth in the 10-20% ADE (with a maximum total weight of 3.26 ± 0.18 g L-1 for C. vulgaris and 2.81 ± 0.10 g L-1 for C. sorokiniana). The use of 10% ADE made it possible to cultivate the strains of the family Scenedesmaceae more effectively than the use of 20% ADE. The growth of Neochloris sp. in ADE was the lowest compared to other microalgal strains. The results of this study demonstrated the feasibility of introducing individual green microalgae into the processes of nutrient recovery from ADE to obtain biomass with a high protein content.

Microalgae/cyanobacteria: the potential green future of vitamin B12 production.

This review summarizes the available information about potential sources of vitamin B12, especially for people who follow a vegan or vegetarian diet and inhabitants of poor countries in the developing world. Cyanobacteria and microalgae approved for food purposes can play a critical role as promising and innovative sources of this vitamin. This work involves a discussion of whether the form of vitamin B12 extracted from microalgae/cyanobacteria is biologically available to humans, specifically focusing on the genera Arthrospira and Chlorella. It describes analyses of their biomass composition, cultivation requirements, and genetic properties in B12 production. Furthermore, this review discusses the function of cobalamin in microalgae and cyanobacteria themselves and the possibility of modification and cocultivation to increase the content of B12 in their biomass.

Fractionation of the water insoluble part of the heterotrophic mutant green microalga Parachlorella kessleri HY1 (Chlorellaceae) biomass: Identification and structure of polysaccharides.

The water-insoluble part of Parachlorella kessleri HY1 biomass was subjected to the extraction of cell-wall polysaccharides using polar aprotic solvents (DMSO, LiCl/DMSO) and aqueous alkaline solutions (0.1, 1 and 4 mol·l-1 of NaOH). Proteins predominated in all the crude extracts and in the insoluble residues were partially removed by treatment with proteolytic enzymes (pepsin and pronase), and in some cases with the HCl/H2O2 reagent, yielding purified polysaccharide-enriched fractions. These treatments led to the solubilisation of some products in water. The composition and structure of isolated polysaccharides were characterised based on monosaccharide composition, glycosidic linkage and spectroscopic analyses. The DMSO extract contained mainly proteins, and polysaccharides were not detected. The water-soluble parts isolated from the LiCl/DMSO extract contained α-l-rhamnan, α-d-glucan and ß-d-glucogalactan; the water-insoluble part contained (1 â†’ 4)-ß-d-xylan, first isolated from the biomass of green microalgae. The alkali extracts contained polysaccharides of similar structure, and also water-insoluble (1 â†’ 4)-ß-d-mannan. The insoluble part after all extractions contained α-chitin as the main polysaccharide, which was confirmed by spectroscopic methods. All these polysaccharides can play a certain role in the cell wall structure of this microalga.

Visualising the Emerging Platform of Using Microalgae as a Sustainable Bio-Factory for Healthy Lipid Production through Biocompatible AIE Probes.

Nowadays, a particular focus is using microalgae to get high-valued health beneficiary lipids. The precise localisation of the lipid droplets (LDs) and biochemical changes are crucial to portray the lipid production strategy in algae, but it requires an in vivo tool to rapidly visualise LD distribution. As a novel strategy, this study focuses on detecting lipid bioaccumulation in a green microalga, Chlamydomonas reinhardtii using the aggregation-induced emission (AIE) based probe, 2-DPAN (C24H18N2O). As the messenger molecule and stress biomarker, hydrogen peroxide (H2O2) activity was detected in lipid synthesis with the AIE probe, TPE-BO (C38H42B2O4). Distinctive LDs labelled with 2-DPAN have elucidated the lipid inducing conditions, where more health beneficiary α-linolenic acid has been produced. TPE-BO labelled H2O2 have clarified the involvement of H2O2 during lipid biogenesis. The co-staining procedure with traditional green BODIPY dye and red chlorophyll indicates that 2-DPAN is suitable for multicolour LD imaging. Compared with BODIPY, 2-DPAN was an efficient sample preparation technique without the washing procedure. Thus, 2-DPAN could improve traditional fluorescent probes currently used for lipid imaging. In addition, the rapid, wash-free, multicolour AIE-based in vivo probe in the study of LDs with 2-DPAN could advance the research of lipid production in microalgae.

Species delimitation and microalgal cryptic diversity analysis of the genus Micractinium (Chlorophyta).

In this article, the system of the green microalgal genus Micractinium, based on morphological, physiological, ecological and molecular data, is considered. The main diagnostic species characteristics and the taxonomic placement of some taxa are also discussed. Phylogenetic analysis showed that the genus Micractinium is characterized by high cryptic diversity. The algorithms used for species delimitation had different results on the number of potentially species-level clusters allocated. The ABGD method was less "sensitive". The tree-based approaches GMYC and PTP showed a more feasible taxonomy of the genus Micractinium, being an effective additional tool for distinguishing species. The clustering obtained by the latter two methods is in good congruence with morphological (cell size and shape, ability to form colonies, production of bristles, chloroplast type), physiological (vitamin requirements, reaction to high and low temperatures), molecular (presence of introns, level of genetic differences, presence of CBCs or special features of the secondary structure in ITS1 and ITS2) and ecological characteristics (habitat). The polyphyly of the holotype of the genus M. pusillum as well as M. belenophorum is shown. The intron was effective as an additional tool for distinguishing species, and the results of the intron analysis should be taken into account together with other characteristics. The CBC approach, based on the search for compensatory base changes in conservative ITS2 regions, was successful only for distinguishing cryptic species from "true" members of M. pusillum. Therefore, to distinguish species, it is more effective to take into account all the CBC in ITS1 and ITS2 and analyze characteristic structural differences (molecular signatures) in the secondary structure of internal transcribed spacers. The genetic distances analysis of 18S-ITS1-5.8S-ITS2 nucleotide sequences showed that intraspecific differences in the genus ranged from 0 to 0.5 % and interspecific differences, from 0.6 to 4.7 %. Due to the polyphasic approach, it was possible to characterize 29 clusters and phylogenetic lines at the species level within the genus Micractinium and to make assumptions about the species.

Cultivable microalgae diversity from a freshwater aquaculture filtering system and its potential for polishing aquaculture-derived water streams.

AIMS: Microalgae are ubiquitous in aquatic environments, including aquaculture farms, but few studies have delved into their phytoplankton diversity and bioremediation potential. In this study, the cultivable phytoplankton of a rainbow trout freshwater aquaculture farm was isolated, phylogenetically analysed and used to assemble a consortium to polish an aquaculture-derived effluent, with low concentrations of ammonium, nitrite and nitrate. METHODS AND RESULTS: Through standard plating in different selective media, a total of 15 microalgae strains were isolated from sludge from a rotary drum filtering system which removes suspended solids from the water exiting the facility. Based on 18S rRNA gene sequences, isolates were assigned to nine different genera of the Chlorophyta phylum: Asterarcys, Chlorella, Chloroccocum, Chlorosarcinopsis, Coelastrella, Desmodesmus, Micractinium, Parachlorella and Scenedesmus. Species from most of these genera are known to inhabit freshwater systems in Galicia and continental Spain, but the Coelastrella, Asterarcys or Parachlorella genera are not usually present in freshwater streams. In an onsite integrative approach, the capacity of a consortium of native microalgae isolates to grow on aquaculture-derived effluents and its nutrient removal capacity were assessed using a raceway pond. After 7 days, removal efficiencies of approximately 99%, 92% and 49% for ammonium, nitrite and nitrate, respectively, were achieved concomitantly with a microalgae biomass increase of ca. 17%. CONCLUSIONS: Sludge from the aquaculture filtering system presents a high diversity of microalgae species from the Chlorophyta phylum, whose application in a consortial approach revealed to be efficient to polish aquaculture-derived effluents with low nutrient content. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of native microalgae consortia from aquaculture systems can contribute to the development of efficient treatment systems for low-nutrient wastewater, avoiding nutrients release to the environment and promoting water recirculation. This may further strengthen the use of phycoremediation at the industrial scale, as an environment-friendly strategy.

Bioaccessibility of microalgae-based carotenoids and their association with the lipid matrix.

The composition of microalgae can contribute to nutritious and functional diets. Among the functional compounds, carotenoids are in focus since positive effects on human health have been established, which are in turn related to their bioaccessibility. In addition to essential nutrients, our hypothesis was that microalgae biomasses could be used as sources of bioaccessible carotenoids. Thus, this study determined for the first time the bioaccessibility of carotenoids from biomass of Scenedesmus bijuga and Chlorella sorokiniana and their possible relationship with the lipid composition of the matrix. The samples were submitted to in vitro digestion protocol, and carotenoids were determined by HPLC-PDA-MS/MS. Individual bioaccessibility of carotenoids was ≥ 3.25%. In general, compounds in their cis conformation were more bioaccessible than trans; and total carotenes more than total xanthophylls. Twelve compounds were bioaccessible from the biomass of S. bijuga, and eight in C. sorokiniana. In S. bijuga, the bioaccessibility of total carotenoids was 7.30%, and the major bioaccessible carotenoids were 9-cis-ß-carotene (43.78%), 9-cis-zeaxanthin (42.30%) followed by 9-cis-lutein (26.73%); while in C. sorokiniana, the total bioaccessibility was 8.03%, and 9-cis-ß-carotene (26.18%), all-trans-ß-carotene (13.56%), followed by 13-cis-lutein (10.71%) were the major compounds. Overall, the total content of lipids does not influence the bioaccessibility of total carotenoids. Still, the lipid composition, including structural characteristics such as degree of saturation and chain length of the fatty acid, impacts the promotion of individual bioaccessibility of carotenes and xanthophylls of microalgae. Finally, the results of this study can assist the development of microalgae-based functional food ingredients and products.

Exploiting the Potential in Water Cleanup from Metals and Nutrients of Desmodesmus sp. and Ampelodesmos mauritanicus.

Increasing levels of freshwater contaminants, mainly due to anthropogenic activities, have resulted in a great deal of interest in finding new eco-friendly, cost-effective and efficient methods for remediating polluted waters. The aim of this work was to assess the feasibility of using a green microalga Desmodesmus sp., a cyanobacterium Nostoc sp. and a hemicryptophyte Ampelodesmos mauritanicus to bioremediate a water polluted with an excess of nutrients (nitrogen and phosphorus) and heavy metals (copper and nickel). We immediately determined that Nostoc sp. was sensitive to metal toxicity, and thus Desmodesmus sp. was chosen for sequential tests with A. mauritanicus. First, A. mauritanicus plants were grown in the 'polluted' culture medium for seven days and were, then, substituted by Desmodesmus sp. for a further seven days (14 days in total). Heavy metals were shown to negatively affect both the growth rates and nutrient removal capacity. The sequential approach resulted in high metal removal rates in the single metal solutions up to 74% for Cu and 85% for Ni, while, in the bi-metal solutions, the removal rates were lower and showed a bias for Cu uptake. Single species controls showed better outcomes; however, further studies are necessary to investigate the behavior of new species.

Strategic evaluation of limiting factors affecting algal growth - An approach to waste mitigation and carbon dioxide sequestration.

This work outlines major critical physico-chemical parameters that play a key role in increasing the fixation of CO2 from coal-fired flue gas CO2 into green microalgae biomass. Nitrogen concentration, gas flow rate, initial medium pH, and incident light intensity were determined to be the most important process variables with significant impact on CO2 fixation. Therefore, NaNO3 (500-3000 mg L-1), pH (6.8-8.0), light (50-200 mol m-2 s-1) and aeration (0.1-1.0 vvm) were varied to assess the biological assimilation potential of CO2 from the flue gas. The parameters that resulted in maximal CO2 fixation from raw flue gas, resulting in a maximum biomass density of 3.1 g L-1, were NaNO3 = 1500 mg L-1, pH =7.2-7.5, incident light intensity = 133.33 mol m-2 s-1, and 0.5-0.75 vvm aeration without any cost-incurring flue gas pre-treatment step. The inductively coupled plasma-mass spectrometer (ICP-MS) was used to investigate heavy metals uptake from raw flue gas, and it was discovered that no net intake of trace metals had a significant influence on biomass production. The research lays the path for efficient large-scale microalgal cultivations for industrial uses, as well as bolstering the circular economy concept.

Comparative responses of cell growth and related extracellular polymeric substances in Tetraselmis sp. to nonylphenol, bisphenol A and 17α-ethinylestradiol.

Estuarine ecosystems near mega-cities are sinks of anthropogenic endocrine disrupting chemicals (EDCs). As the most important primary producer, indigenous microalgae and their secreted extracellular polymeric substances (EPSs) might interact with EDCs and contribute to their fate and risk. Tetraselmis sp. is a representative model of estuarine microalga, for which EDC toxicity and its effects on EPS synthesis have rarely been studied. Through microalgal isolation, algal cell growth tests, EDC removal and the characterization of related EPS profiles, the present work intends to clarify the comparative responses of Tetraselmis sp. to nonylphenol (NP), bisphenol A (BPA) and 17α-ethinylestradiol (EE2). The results showed that the half inhibitory concentration on cell growth was 0.190-0.313 mg/dm3 for NP, which was one order of magnitude lower than the comparable values for BPA and EE2 at 2.072-3.254 mg/dm3. Regarding chlorophyll, NP induced its degradation, EE2 led to its decreased production, and BPA had no obvious effect. Under EDC stress, only the concentrations of colloidal polysaccharides and proteins responded dose-dependently to EE2. Except for the colloidal fraction in the EE2 treatment group, the increase in neutral monosaccharides, especially glucose and galactose, was a common response to EDCs. Compared to the recalcitrant BPA, NP underwent abiotic degradation in alga-free water, and EE2 could be biodegraded in water containing this microalga. The chemical-specific responses of cell growth, chlorophyll and related EPS profiles were driven by the different fates of EDCs, and the underlying mechanism was further discussed. The results obtained in the present work are of critical importance for understanding the fate and effects of different EDCs mediated by microalgae and their related EPSs.

Selection and re-acclimation of bioprospected acid-tolerant green microalgae suitable for growth at low pH.

For mass culture of photosynthetic green microalgae, industrial flue gases can represent a low-cost resource of CO2. However, flue gases are often avoided, because they often also contain high levels of SO2 and/or NO2, which cause significant acidification of media to below pH 3 due to production of sulfuric and nitric acid. This creates an unsuitable environment for the neutrophilic microalgae commonly used in large-scale commercial production. To address this issue, we have looked at selecting acid-tolerant microalgae via growth at pH 2.5 carried out with samples bioprospected from an active smelter site. Of the eight wild samples collected, one consisting mainly of Coccomyxa sp. grew at pH 2.5 and achieved a density of 640 mg L-1. Furthermore, three previously bioprospected green microalgae from acidic waters (pH 3-4.5) near abandoned mine sites were also re-acclimated down to their in-situ pH environment after approximately 4 years spent at neutral pH. Of those three, an axenic culture of Coccomyxa sp. was the most successful at re-acclimating and achieved the highest density of 293.1 mg L-1 and maximum daily productivity of 38.8 mg L-1 day-1 at pH 3. Re-acclimation of acid-tolerant species is, therefore, achievable when directly placed at their original pH, but gradual reduction in pH is recommended to give the cells time to acclimate.