Microalgal Cells Research Articles

Futuristic opportunities for pretreatment processes in biofuel production from microalgae

AbstractMicroalgal biofuel is a promising solution to replace fossil fuel as a renewable and environmental‐friendly energy source, thereby contributing to the United Nations (UN) Sustainable Development Goals (SDGs), in particular SDG‐7, or Affordable and Clean Energy. Unlike energy crops (like oil palm and sugar cane), microalgae benefit from faster growth rate, higher lipid content, smaller land area required, ability to flourish using waste or brackish water, and posing zero competition with food crops. Microalgae‐derived biofuels (like biodiesel, bioethanol, biomethane, and biohydrogen) are sustainable energy sources that can be produced using well‐developed techniques (e.g., transesterification, fermentation, anaerobic digestion, and Fisher–Tropsch process). To prevent dire climate conditions resulting from the global temperature rise of 1.5°C and resolve worldwide energy security issue, our generation will need to establish and implement renewables on a global scale. To improve the industrial production of microalgal biofuel, the efficiencies of biomass and metabolite production to post‐cultivation biofuel synthesis processes must be enhanced. For the cultivation step, there exist three key techniques that can directly change the traits, structure, and behavior of microalgal cells, and induce them to accumulate targeted metabolites rapidly and in large amounts. These techniques are genetic engineering, chemical modulation, and nanomaterial approach. Genetic engineering commonly alters the chloroplast DNA of microalgae to overexpress or down‐regulate key genes in various metabolic pathways so that the cells accumulate more lipids. Chemicals can also be used to modulate microalgal growth and lipid accumulation by inducing oxidative stress or prevent conversion of lipid molecules. Nanomaterials and nanoparticles can also enhance microalgal lipid production by microenvironmental stress induction, vitamin supplementation, and light backscattering. Therefore, in this review, the recent progress as well as the pros and cons of genetic engineering, chemical modulation, and nanomaterial approach in achieving greater biofuel production from microalgae are comprehensively examined.

Inhibitory effect of malonic acid on Microcystis aeruginosa: Cell morphology, oxidative stress and gene expression

The utilization of allelochemicals, which are regarded as eco-friendly algaecides for mitigating harmful cyanobacterial blooms (HCB), has attracted increasing attention, owing to their negligible toxicity, high selectivity and excellent degradability. In the present study, the inhibitory effects of malonic acid on Microcystis aeruginosa were investigated in terms of growth indicators, cell morphology, antioxidant enzymes activities and gene expression. The results indicated that the exposure to 0.5 mmol L−1 malonic acid for 7 d caused a high microalgae inhibition rate of 86.88 %. Furthermore, the analysis of scanning electron microscopy (SEM) and transmission electron microscope (TEM) showed that the presence of malonic acid resulted in the disruption of cell integrity and the changes in cell morphology. Specifically, malonic acid was observed to cleave microalgae cells, while concurrently oxidizing the microalgae organic matter. In addition, under the exposure of 0.5 mmol L−1 malonic acid, the levels of malondialdehyde (MDA) were 3.28 times as the control group on d-5, while the activities of superoxide dismutase (SOD) and catalase (CAT) were first increased by 5.96 and 11.73 fold compare to the control group on d-5, and then decreased, due to oxidative stress induced by malonic acid. Transcriptome-based gene expression analysis revealed that the presence of malonic acid significantly inhibited the transcription of genes (rbcL, psbA1) associated with photosynthesis, and the expression of DNA repair gene (recA), stress response gene (ftsH) and microcystin synthesis genes (mcyD, mcyH) were up-regulated. These findings provide novel insights into the inhibitory effect of malonic acid on M. aeruginosa, suggesting that malonic acid has the potential to control M. aeruginosa blooms.

Impact of organic matter molecular weight on hexavalent chromium enrichment in green microalgae

In lightly polluted water containing heavy metals, organic matter, and green microalgae, the molecular weight of organic matter may influence both the growth of green microalgae and the concentration of heavy metals. This study elucidates the effects and mechanisms by which different molecular weight fractions of fulvic acid (FA), a model dissolved organic matter component, facilitate the bioaccumulation of hexavalent chromium (Cr(VI)) in a typical green alga, Chlorella vulgaris. Findings show that the addition of FA fractions with molecular weights greater than 10 kDa significantly enhances the enrichment of total chromium and Cr(VI) in algal cells, reaching 21.58%−31.09 % and 16.17 %−22.63 %, respectively. Conversely, the efficiency of chromium enrichment in algal cells was found to decrease with decreasing molecular weight of FA. FA molecular weight within the range of 0.22 µm-30 kDa facilitated chromium enrichment primarily through the algal organic matter (AOM) pathway, with minor contributions from the algal cell proliferation and extracellular polymeric substances (EPS) pathways. However, with decreasing FA molecular weight, the AOM and EPS pathways become less prominent, whereas the algal cell proliferation pathway becomes dominant. These findings provide new insights into the mechanism of chromium enrichment in green algae enhanced by medium molecular weight FA.

Roles of extracellular polymeric substances in arsenic accumulation and detoxification by cell wall intact and mutant strains of Chlamydomonas reinhardtii

Arsenic (As) pollution seriously threatens human and ecological health. Microalgal cell wall and extracellular polymeric substances (EPS) are known to interact with As, but their roles in the As resistance, accumulation and speciation in microalgae remain unclear. Here, we used two strains of Chlamydomonas reinhardtii, namely CC-125 (wild type) and CC-503 (cell wall-deficient mutant), to examine the algal growth, EPS synthesis, As adsorption, absorption and transformation under 10–1000 µg/L As(III) and As(V) treatments for 96 h. In both strains, the As absorption increased after the EPS removal, but the growth, As adsorption, and transformation of C. reinhardtii declined. The CC-125 strain was more tolerant to As stress and more efficient in EPS production, As accumulation, and redox transformation than CC-503, irrespective of EPS presence or absence. Three-dimension excitation-emission matrix (3D-EEM) and attenuated total reflectance infrared spectroscopy (ATR-IR) analyses showed that As was bound with functional groups in the EPS and cell wall, such as -COOH, NH and -OH in proteins, polysaccharides and amino acids. Together, this study demonstrated that EPS and cell wall acted as barriers to lower the As uptake by C. reinhardtii. However, the cell wall mutant strain was more susceptible to As toxicity due to lower EPS induction and higher As absorption.

Natural deep eutectic solvents (NADES) for the extraction of bioactives: emerging opportunities in biorefinery applications

Natural deep eutectic solvents (NADES) have emerged as an eco-friendly alternative for extracting bioactives, avoiding the use of flammable organic solvents and extreme temperatures and pH conditions. NADES rely on intermolecular interactions between hydrogen bonding donors (HBD) and hydrogen bonding acceptors (HBA) to form eutectic mixtures with significantly lower melting points than their individual components. These matrices are influenced by factors like water content, temperature, and component ratios. NADES high viscosity can hinder extractive efficiency, which can be mitigated by adding water or working at higher temperatures. However, excessive dilution with water may disrupt the supramolecular structure of NADES, reducing extraction efficiency. A notable feature of NADES is their fine-tunability for specific purposes. Adjusting physicochemical properties such as polarity, pH, and viscosity optimizes extraction efficiency by promoting the solubility of target molecules and interactions between the NADES and target molecules. NADES, unlike organic solvents, can partially disrupt plant and microalgae cell walls, enhancing permeability and extraction efficiency. Moreover, NADES can have a stabilising effect on bioactives and can enhance their biological activity and bioavailability. These attributes, coupled with their low environmental impact in terms of low toxicity and high biodegradability, make NADES attractive for biorefinery applications.

Antibiotic study of non-polar microalgae extract <i>Chlorella sorokiniana</i> against gram-positive bacteria

A study of the antibacterial properties of a non-polar extract of microalgae Chlorella sorokiniana on gram-positive bacteria is presented along with a determination of the minimum inhibitory concentrations of the mixture and the individual metabolites that make up the extract. A regular effect of illumination on the intensity of the antibiotic effect of non-polar microalgae extract on gram-positive bacteria is demonstrated. A mixture of substances extracted from disintegrated cells of the microalgae biomass Chlorella sorokiniana has an inhibitory effect on bacterial growth at a photosynthetically active radiation level of 100±6 μmol photons/(m2×s). The minimum effective amount of the extract is 330±11.09 µg. When analysing the chemical structure of the components of the non-polar fraction extracted from the cells of microalgae Chlorella sorokiniana, the composition of the non-polar extract was shown to include triacylglycerides, fatty acids, o-dialkyl monoglycerides and ethers of sterols or waxes, or trialkyl esters of glycerol. When studying the antibiotic properties of individual fractions of substances, triacylglycerides and fatty acids were found to have an antibiotic effect on gram-positive bacteria. In this case, the minimum effective amount of triacylglycerides is 400±13.37 μg, while that of fatty acids is 600±20.05 μg. The combined effect of a mixture of non-polar extract substances gives the most pronounced antibiotic effect on gram-positive bacteria at a photosynthetically active radiation level of 100±6 μmol of photons/(m2×s). Thus, an increase in antibacterial action was demonstrated when using a mixture of substances of the non-polar extract of microalgae Chlorella sorokiniana at a photosynthetically active radiation level of 100±6 μmol of photons/(m2×s).

Reactive oxygen species mediated extracellular polymeric substances production assisting the recovery of Thalassiosira pseudonana from polystyrene micro and nanoplastics exposure

As emerging pollutants in the aquatic environments, micro- and nano-plastics (MNPs) aroused widespread environmental concerns for their potential threats to the ecological health. Previous research has proved that microalgae growth could recover from the MNPs toxicities, in which the extracellular polymeric substances (EPS) might play the key role. In order to comprehensively investigate the recovery process of microalgae from MNPs stress and the effecting mechanisms of EPS therein, this study conducted a series of experiments by employing two sizes (0.1 and 1 μm) of polystyrene (PS) MNPs and the marine model diatom Thalassiosira pseudonana during 14 days. The results indicated: the pigments accumulations and photosynthetic recovery of T. pseudonana under MPs exposure showed in the early stage (4–5 days), while the elevation of reactive oxygen species (ROS) and EPS contents lasted longer time period (7–8 days). EPS was aggregated with MNPs particles and microalgal cells, corresponding to the increased settlement rates. More increase of soluble (SL)-EPS contents was found than bound (B)-EPS under MNPs exposure, in which the increase of the protein proportion and humic acid-like substances in SL-EPS was found, thus facilitating aggregates formation. ROS was the signaling molecule mediating the overproduction of EPS. The transcriptional results further proved the enhanced EPS biosynthesis on the molecular level. Therefore, this study elucidated the recovery pattern of microalgae from MNPs stress and linked “ROS-EPS production changes-aggregation formation” together during the growth recovery process, with important scientific and environmental significance.

Aquatic Fate and Ecotoxicology Effect of ZnS:Mn Quantum Dots on Chlorella vulgaris in Fresh Water.

With the increasing integration of nanomaterials into daily life, the potential ecotoxicological impacts of nanoparticles (NPs) have attracted increased attention from the scientific community. This study assessed the ecotoxicity of ZnS quantum dots (QDs) doped with varying molar concentrations of Mn2+ on Chlorella vulgaris. The ZnS:Mn QDs were synthesized using the polyol method. The size of the ZnS:Mn QDs ranged from approximately 1.1 nm to 2 nm, while the aggregation size in Seine River water was 341 nm at pH 6 and 8. The presence of ZnS:Mn (10%) NPs exhibited profound toxicity to Chlorella vulgaris, with immediate reductions in viability (survival cells) from 71%, 60% to 51%, 52% in BG11 and Seine River water, respectively, at a concentration of 100 mg L-1 of ZnS:Mn (10%) NPs. Additionally, the ATP content in Chlorella vulgaris significantly decreased in Seine River water (by 20%) after 3 h of exposure to ZnS:Mn (10%) NPs. Concurrently, SOD activity significantly increased in Seine River water, indicating that the ZnS:Mn (10%) NPs induced ROS production and triggered an oxidative stress response in microalgae cells.

Mitigating salinity stress through interactions between microalgae and different forms (free-living & alginate gel-encapsulated) of bacteria isolated from estuarine environments

Salinity stress in estuarine environments poses a significant challenge for microalgal survival and proliferation. The interaction between microalgae and bacteria shows promise in alleviating the detrimental impacts of salinity stress on microalgae. Our study investigates this interaction by co-cultivating Chlorella sorokiniana, a freshwater microalga, with a marine growth-promoting bacterium Pseudomonas gessardii, both of which were isolated from estuary. In this study, bacteria were encapsulated using sodium alginate microspheres to establish an isolated co-culture system, preventing direct exposure between microalgae and bacteria. We evaluated microalgal responses to different salinities (5 PSU, 15 PSU) and interaction modes (free-living, gel-encapsulated), focusing on growth, photosynthesis, cellular metabolism, and extracellular polymeric substances (EPS) properties. High salinity inhibited microalgal proliferation, while gel-fixed interaction boosted Chlorella growth rate by 50.7 %. Both attached and free-living bacteria restored Chlorella's NPQ to normal levels under salt stress. Microalgae in the free-living interaction group exhibited a significantly lower respiratory rate compared to the pure algae group (−17.2 %). Increased salinity led to enhanced EPS polysaccharide secretion by microalgae, particularly in interaction groups (19.7 %). Both salt stress and interaction increased the proportion of aromatic proteins in microalgae's EPS, enhancing its stability by modulating EPS glycosidic bond C-O-C and protein vibrations. This alteration caused microalgal cells to aggregate, free-living bacteria co-culture group, and fixed co-culture group increasing by 427.5 %, 567.1 %, and 704.1 %, respectively. In gel-fixed bacteria groups, reduced neutral lipids don't accumulate starch instead, carbon redirects to cellular growth, aiding salt stress mitigation. These synergistic activities between salinity and bacterial interactions are vital in mitigating salinity stress, improving the resilience and growth of microalgae in saline conditions. Our research sheds light on the mechanisms of microalgal-bacterial interactions in coping with salt stress, offering insights into the response of estuarine microorganisms to global environmental changes and their ecological stability.

Toxicity of urban stormwater on Chlorella pyrenoidosa: Implications for reuse safety

Urban stormwater is an alternative water source used to mitigate water resource shortages, and ensuring the safety of stormwater reuse is essential. An in-depth understanding of both individual pollutant concentrations/loads in stormwater and holistic stormwater quality can be used to comprehensively evaluate how safely stormwater can be reused. The toxicity test takes all pollutants present in water samples into account, and the results reflect the integrated effect of these pollutants. In this study, the influence of urban stormwater sourced from different land uses on microalgae (Chlorella pyrenoidosa) and the possible toxicity mechanisms were investigated. The results showed that urban stormwater, particularly residential road stormwater, significantly inhibited microalgal growth. The chlorophyll contents of microalgae exposed to residential road stormwater were relatively lower, while the corresponding values were relatively higher for microalgae exposed to grassland road stormwater. Additionally, the antioxidant-related metabolism of microalgae could be dysregulated due to exposure to urban stormwater. A possible toxicity mechanism is that urban stormwater influences metabolic pathways related to chlorophyll synthesis and further hinders photosynthesis and hence microalgal growth. To resist oxidative stress and maintain regular microalgal cell activities, the ribosome metabolism pathway was upregulated. The research results contribute to elucidating the toxicity effects of urban stormwater and hence provide useful insight for ensuring the safety of stormwater reuse. It is also worth noting that the study outcomes can only represent the influence of land use on stormwater toxicity, while the impacts of other factors (particularly rainfall-runoff characteristics) have not been considered. Therefore, the consideration of all influential factors of stormwater is strongly recommended to generate more robust results in the future and provide more effective guidance for real practices related to stormwater reuse safety.

Effects of HCO3− and CO2 conversion rates on carbon assimilation strategies in marine microalgae: Implication by stable carbon isotope analysis of fatty acids

Marine microalgae are an essential component of marine plankton and critical primary producers, playing a vital role in marine ecosystems. The seawater carbonate system is a dynamic equilibrium system, and changes in any component can alter the carbonate balance. In CO2-concentrating mechanisms (CCMs), carbonic anhydrase (CA) regulates CO2 concentration by catalyzing the interconversion between CO2 and HCO3−. Therefore, limiting the activity of extracellular carbonic anhydrase (exCA) alters the rate at which carbonate equilibrium is reached and further affects the carbon assimilation process in microalgae. In this study, two different microalgae, Phaeodactylum tricornutum and Nannochloropsis oceanica, were selected to investigate the effects of changes in the carbonate system on photosynthetic carbon assimilation in microalgae by inhibiting exCA activity with acetazolamide (AZ). Inhibition of exCA activity reduces specific growth rates and photosynthetic efficiency of microalgae. The total alkalinity, HCO3− concentration, and CO2 concentration of the cultures increased with the decrease of pH, but the changes of the ribulose 1,5- bisphosphate carboxylase/oxygenase (Rubisco) activities of the two microalgae were different. In addition, the two microalgae possessed different lipid and carbohydrate synthesis strategies, but both restricted triacylglycerol (TAG) synthesis. Meanwhile, the microalgal cells had to utilize more 13CO2 when HCO3− and CO2 conversion rates were limited and restricted. This led to the continuous accumulation of 13C in fatty acids and the elevation of δ13CFAs. In conclusion, our study provides a new perspective on the role of microalgae in correlating carbonate changes with photosynthetic carbon assimilation strategies under mechanistic constraints on inorganic carbon utilization.

Valorization of Nannochloropsis oceanica for integrated co-production of violaxanthin cycle carotenoids

The development of integrated co-production of multiple high-purity carotenoids from microalgal cells holds considerable significance for the valorization of microalgae. In this study, the economical microalga Nannochloropsis oceanica was identified as an accumulator of violaxanthin cycle carotenoids, including violaxanthin, antheraxanthin, and zeaxanthin. Notably, a novel and competent approach for the integrated co-production of violaxanthin cycle carotenoids was explored, encompassing four steps: microalgal cultivation, solvent extraction, octadecylsilyl open-column chromatography, and ethanol precipitation. Under optimal co-production conditions, the purities of the obtained violaxanthin, antheraxanthin, and zeaxanthin all exceeded 92%, with total recovery rates of approximately 51%, 40%, and 60%, respectively. Utilizing nuclear magnetic resonance techniques, the purified violaxanthin, antheraxanthin, and zeaxanthin were identified as all-trans-violaxanthin, all-trans-antheraxanthin, and all-trans-zeaxanthin, respectively. This method held significance for the multiproduct biorefinery of the microalga N. oceanica and carried potential future implications for the violaxanthin cycle carotenoids.

Effects of chemical oxygen demand and chloramphenicol on attached microalgae growth: Physicochemical properties and microscopic mass transfer in biofilm

During the wastewater treatment and resource recovery process by attached microalgae, the chemical oxygen demand (COD) can cause biotic contamination in algal culture systems, which can be mitigated by adding an appropriate dosage of antibiotics. The transport of COD and additive antibiotic (chloramphenicol, CAP) in algal biofilms and their influence on algal physiology were studied. The results showed that COD (60 mg/L) affected key metabolic pathways, such as photosystem II and oxidative phosphorylation, improved biofilm autotrophic and heterotrophic metabolic intensities, increased nutrient demand, and promoted biomass accumulation by 55.9 %, which was the most suitable COD concentration for attached microalgae. CAP (5–10 mg/L) effectively stimulated photosynthetic pigment accumulation and nutrient utilization in pelagic microalgal cells. In conclusion, controlling the COD concentration (approximately 60 mg/L) in the medium and adding the appropriate CAP concentration (5–10 mg/L) are conducive to improving attached microalgal biomass production and resource recovery potential from wastewater.

Marine Heat Waves in the Indian Ocean: A Major Climate Impact on Marine Microalgae

A clearly defined, lengthy (five days or longer) period of extraordinarily high ocean temperatures is known as Marine Heat Waves (MHWs) (above the e0th percentile). These events significantly impact the structure, abundance, cell size, metabolic functions, physico-chemical interactions between microalgae cells and their surroundings in the Indian Ocean. In the tropical Indian Ocean, the average sea surface temperature (SST) increased by 1.0°C (0.15°C/decade) on average between 1951 and 2015, while the global average SST warmed by roughly 0.7°C (0.11 °C/decade). This resulted from the incidence of Marine microalgal bloom MMBs (including Harmful Algal Bloom) and the number of causal taxa may grow as SST increases. Trichodesmium and Noctiluca contributed 34.4% and 31.8%, respectively, to the overall blooms. Such a heating phenomenon also triggers the dominance of small-sized algal cells, harmful diatoms and pico-plankton. The present paper reviews the Indian as well as global episodes of MHWs such as Western South Atlantic, North-East Pacific, Western Australia, Alaskan Sea etc. and their impact on phytoplankton. The combination of warming and heatwaves triggers blooms of buoyant cyanobacteria and its combined impacts of MHWs and ocean warming/acidification on phytoplankton community structure have not been fully studied in the Indian Ocean.

Enhancing lipid production and sedimentation of Chlorella pyrenoidosa in saline wastewater through the addition of agricultural phytohormones

In this study, the effect of external agricultural phytohormones (mixed phytohormones) addition (1.0, 5.0, 10.0, and 20.0 mg L−1) on the growth performance, lipid productivity, and sedimentation efficiency of Chlorella pyrenoidosa cultivated in saline wastewater was investigated. Among the different concentrations evaluated, the highest biomass (1.00 g L−1) and lipid productivity (11.11 mg L−1 d−1) of microalgae were obtained at 10.0 mg L−1 agricultural phytohormones addition. Moreover, exogenous agricultural phytohormones also improved the sedimentation performance of C. pyrenoidosa, which was conducive to the harvest of microalgae resources, and the improvement of sedimentation performance was positively correlated with the amount of agricultural phytohormones used. The promotion of extracellular polymeric substances synthesis by phytohormones in microalgal cells could be considered as the reason for its promotion of microalgal sedimentation. Transcriptome analysis revealed that the addition of phytohormones upregulated the expression of genes related to the mitogen-activated protein kinase (MAPK)-mediated phytohormone signaling pathway and lipid synthesis, thereby improving salinity tolerance and lipid production in C. pyrenoidosa. Overall, agricultural phytohormones provide an effective and inexpensive strategy for increasing the lipid productivity and sedimentation efficiency of microalgae cultured in saline wastewater.

Modelling photoperiod in enhancing hydrogen production from Chlorella vulgaris sp. while bioremediating ammonium and organic pollutants in municipal wastewater

Municipal wastewater is ubiquitously laden with myriad pollutants discharged primarily from a combination of domestic and industrial activities. These heterogeneous pollutants are threating the natural environments when the traditional activated sludge system fails sporadically to reduce the pollutants' toxicities. Besides, the activated sludge system is very energy intensive, bringing conundrums for decarbonization. This research endeavoured to employ Chlorella vulgaris sp. In converting pollutants from municipal wastewater into hydrogen via alternate light and dark fermentative process. The microalgae in attached form onto 1 cm3 of polyurethane foam cubes were adopted in optimizing light intensity and photoperiod during the light exposure duration. The highest hydrogen production was recorded at 52 mL amidst the synergistic light intensity and photoperiod of 200 μmolm−2s−1 and 12:12 h (light:dark h), respectively. At this lighting condition, the removals of chemical oxygen demand (COD) and ammoniacal nitrogen were both achieved at about 80%. The sustainability of microalgal fermentative performances was verified in recyclability study using similar immobilization support material. There were negligible diminishments of hydrogen production as well as both COD and ammoniacal nitrogen removals after five cycles, heralding inconsequential microalgal cells’ washout from the polyurethane support when replacing the municipal wastewater medium at each cycle. The collected dataset was finally modelled into enhanced Monod equation aided by Python software tool of machine learning. The derived model was capable to predict the performances of microalgae to execute the fermentative process in producing hydrogen while subsisting municipal wastewater at arbitrary photoperiod. The enhanced model had a best fitting of R2 of 0.9857 as validated using an independent dataset. Concisely, the outcomes had contributed towards the advancement of municipal wastewater treatment via microalgal fermentative process in producing green hydrogen as a clean energy source to decarbonize the wastewater treatment facilities.

Dialysis bag-microalgae photobioreactor: Novel strategy for enhanced bioresource production and wastewater purification

Cultivating microalgae in wastewater offers various advantages, but it still faces limitations such as bacteria and other impurities in wastewater affecting the growth and purity of microalgae, difficulty in microalgae harvesting, and extracellular products of microalgae affecting effluent quality. In this study, a novel dialysis bag-microalgae photobioreactor (Db-PBR) was developed to achieve wastewater purification and purer bioresource recovery by culturing microalgae in a dialysis bag. The dialysis bag in the Db-PBR effectively captured the microalgae cells and promoted their lipid accumulation, leading to higher biomass (1.53 times of the control) and lipid production (2.50 times of the control). During the stable operation stage of Db-PBR, the average soluble microbial products (SMP) content outside the dialysis bag was 25.83 mg L−1, which was significantly lower than that inside the dialysis bag (185.63 mg L−1), indicating that the dialysis bag effectively intercepted the SMP secreted by microalgae. As a result, the concentration of dissolved organic carbon (DOC) in Db-PBR effluent was significantly lower than that of traditional photobioreactor. Furthermore, benefiting from the dialysis bag in the reactor effectively intercepted the microorganisms in wastewater, significantly improving the purity of the cultured microalgae biomass, which is beneficial for the development of high-value microalgae products.

Methylmercury Effect and Distribution in Two Extremophile Microalgae Strains Dunaliella salina and Coccomyxa onubensis from Andalusia (Spain).

The main entrance point of highly toxic organic Hg forms, including methylmercury (MeHg), into the aquatic food web is phytoplankton, which is greatly represented by various natural microalgal species. Processes associated with MeHg fate in microalgae cells such as uptake, effects on cells and toxicity, Hg biotransformation, and intracellular stability are detrimental to the process of further biomagnification and, as a consequence, have great importance for human health. The study of MeHg uptake and distribution in cultures of marine halophile Dunaliella salina and freshwater acidophilic alga Coccomyxa onubensis demonstrated that most of the MeHg is imported inside the cell, while cell surface adhesion is insignificant. Almost all MeHg is removed from the culture medium after 72 h. Significant processes in rapid MeHg removal from liquid medium are its abiotic photodegradation and volatilization associated with algal enzymatic activity. The maximum intracellular accumulation for both species was in 80 nM MeHg-exposed cultures after 24 h of exposure for D. salina (from 27 to 34 µg/gDW) and at 48 h for C. onubensis (up to 138 µg/gDW). The different Hg intakes in these two strains could be explained by the lack of a rigid cell wall in D. salina and the higher chemical ability of MeHg to pass through complex cell wall structures in C. onubensis. Electron microscopy studies on the ultrastructure of both strains demonstrated obvious microvacuolization in the form of many very small vacuoles and partial cell membrane disruption in 80 nM MeHg-exposed cultures. Results further showed that Coccomyxa onubensis is a good candidate for MeHg-contaminated water reclamation due to its great robustness at nanomolar concentrations of MeHg coupled with its very high intake and almost complete Hg removal from liquid medium at the MeHg levels tested.

Innovative vs classical methods for drying heterotrophic Chlorella vulgaris: Impact on protein quality and sensory properties

Drying is a necessary step in the microalgae production chain to reduce microbial load and oxidative degradation of the end product. Depending on the differences in applied temperature and treatment time, the process of drying can have a substantial impact on protein quality and aroma, important characteristics determining the incorporation potential in food products. In this study, we compared the drying of heterotrophic Chorella vulgaris with both innovative (agitated thin film drying (ATFD), pulse combustion drying (PCD) and solar drying (SolD)) and commonly used drying techniques (spray drying (SprD) and freeze drying (FD)). To evaluate the impact on protein quality, we evaluated techno-functional properties, in vitro digestibility (INFOGEST) as well as protein denaturation using differential scanning calorimetry (DSC). A sensory analysis was performed by a trained expert panel, combined with headspace solid-phase microextraction (HS-SPME) - gas chromatography-mass spectrometry (GC–MS) to determine volatile organic compounds (VOCs). ATFD was found to increase techno-functional properties such as gelling, water holding and solubility as well as in vitro protein digestibility. These observations could be related to induced cell disruption and protein denaturation by ATFD. Sensory analysis indicated an increased earthy off-flavor after ATFD. Interestingly, the high-temperature PCD led to an increase in cacao odor while low-temperature FD resulted in lower flavor, odors and VOCs. These results demonstrate that protein quality and sensorial properties of C. vulgaris can be steered through the type of drying, which could help in the selection of application-specific drying methods. Overall, this work could promote the incorporation of microalgal single cell proteins in different innovative food products.

Fast multispectral imaging analysis as phenotyping platform: Effect of titanium dioxide nanoparticles on microalgae physiology

Several types of nanoparticles have been developed and explored in agricultural applications and their environmental impact evaluated. However, no such study was carried out using fully dispersible ultrasmall titanium dioxide nanoparticles (us-TiO2). Therefore, we investigate the effects of fully dispersible 2.5 nm large us-TiO2 nanoparticles on the cellular metabolism of the microalgae Chlamydomonas reinhardtii using multispectral imaging analysis as a platform for the evaluation of their physiological responses. Our results indicate that us-TiO2 modulates the microalgae cell growth in a concentration-dependent manner. X-ray fluorescence analysis showed that the us-TiO2 strongly interacts with the cells in relatively low and constant amounts in the range of 0.05 g·L−1 to 1.5 g·L−1. Most nanoparticles were found on the cell surface, surrounding the microalgae cells dispersed in TAP medium, indicating a low trend of internalization. We further applied multispectral imaging analyses toward fast phenotyping of the alterations in the cell's physiology. Furthermore, higher concentrations of us-TiO2 (~1 g·L−1) positively affect the microalgae photosynthetic activity and growth. Thus, our results indicated the capacity to detect variations in cell physiology and indicated a possible role of us-TiO2 in modulating positively microalgae photosynthesis and growth in a concentration-dependent way. The mechanisms controlling such responses must be further investigated.