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The addition of metal oxide nanoparticles (NPs) to microalgae cultures have become of great interest since they can directly and/or indirectly interact with microalgae resulting in enhanced production of biomass and high-value added products. The physicochemical characteristics of the NPs, i.e., crystal phase, can differently influence microalgae metabolism, thus the synthesis of optimal NPs is mandatory to improve the value of microalgae biomass.This study aimed at assessing the effect of Fe2O3 NPs on Chlorella spp. cultures with and without carbonate supplementation to confirm their beneficial effect. First, Fe2O3 NPs were synthetized and two different annealing temperatures were implemented, 450 °C (Fe2O3-450) and 1000 °C (Fe2O3-1000), to obtain Fe2O3 NPs with different crystal phases. The NPs crystal phase was analyzed by X-ray diffraction and the presence of the γ- and α- phases on both NPs were confirmed. However, the Fe2O3-450 NPs presented a higher content of the γ- phase. Subsequently, the effect of the crystal phase of Fe2O3 NPs was assessed in Chlorella spp. cultures. The addition of Fe2O3-1000 NPs resulted in inhibition of Chlorella spp. On the other hand, the addition of Fe2O3-450 NPs recorded the carbohydrate enhancements > 91% in Chlorella spp. cultures. The indirect effect of the synthetized NPs was assessed in microalgae cultures without carbonate supplementation and the same tendency was observed. This study proves that Fe2O3-450 NPs directly and indirectly interact with microalgae, and can be used as a strategy to produce high-value biomass even in environments without carbonate supplementation. [ABSTRACT FROM AUTHOR]

Microalgae cultures have emerged as a promising strategy in diverse areas, ranging from wastewater treatment to biofuel production, thus contributing to the search for carbon neutrality. These photosynthetic organisms can utilize the resources present in wastewater and fix atmospheric CO 2 to produce biomass with high energy potential. In this study, the removal efficiency of Polycyclic Aromatic Hydrocarbons (PAHs), CO 2 fixation and lipid content in the biomass produced from microalgae grown in airlift photobioreactor were evaluated. Four mesoscale cultures were carried out: Control (Seawater + Conway medium), Treatment A (Oil Produced Water + Poultry Effluent Water), Treatment B (Poultry Effluent Water + Seawater) and Treatment C (Oil Produced Water, Seawater and nutrients). The impact of biostimulation, through the addition of nutrients, on PAHs removal efficiency (up to 90%), CO 2 fixation rate (up to 0.20 g L -1 d -1 ) and the composition of the generated biomass was observed. Primarily, the addition of nitrates to the culture medium impacted CO 2 fixation rate of the microalgae. In addition, a direct correlation was observed between PAHs removal and lipid accumulation in the biomass, up to 36% in dry weight, demonstrating microalgae's ability to take advantage of the organic carbon (PAHs) present in the culture medium to generate lipid-rich biomass. The concentration of polysaccharides in the biomass obtained did not exceed 12% on a dry weight basis, and the Higher Heating Value (HHV) ranged between 17 and 21 MJ kg -1 . Finally, the potential of generating hydrogen through pyrolysis was highlighted, taking advantage of the characteristics of biomass as a conversion route to produce biofuels. These results show that microalgae are effective in wastewater treatment and have great potential in producing biofuels, thus contributing to the transition towards more sustainable energy sources and climate change mitigation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Aquaculture farming generates a significant amount of wastewater, which has prompted the development of creative bioprocesses to improve wastewater treatment and bioresource recovery. One promising method of achieving these aims is to directly recycle pollutants into microbe-rice bran complexes, which is an economical and efficient technique for wastewater treatment that uses synergetic interactions between algae and bacteria. This study explores novel bioaugmentation as a promising strategy for efficiently forming microbial-rice bran complexes in unsterilized aquaculture wastewater enriched with agricultural residues (molasses and rice bran). Results found that rice bran serves a dual role, acting as both an alternative nutrient source and a biomass support for microalgae and bacteria. Co-bioaugmentation, involving the addition of probiotic bacteria (Bacillus syntrophic consortia) and microalgae consortiums (Tetradesmus dimorphus and Chlorella sp.) to an existing microbial community, led to a remarkable 5-fold increase in microbial-rice bran complex yields compared to the non-bioaugmentation approach. This method provided the most compact biofloc structure (0.50 g/L) and a large particle diameter (404 μm). Co-bioaugmentation significantly boosts the synthesis of extracellular polymeric substances, comprising proteins at 6.5 g/L and polysaccharides at 0.28 g/L. Chlorophyta, comprising 80% of the total algal phylum, and Proteobacteria, comprising 51% of the total bacterial phylum, are emerging as dominant species. These microorganisms play a crucial role in waste and wastewater treatment, as well as in the formation of microbial-rice bran complexes that could serve as an alternative aquaculture feed. This approach prompted changes in both microbial community structure and nutrient cycling processes, as well as water quality. These findings provide valuable insights into the transformative effects of bioaugmentation on the development of microbial-rice bran complexes, offering potential applications in bioprocesses for waste and wastewater management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Microalgae have gained considerable attention as promising candidates for precision nutrition and dietary regulation due to their versatile metabolic capabilities. This review innovatively applies system metabolic engineering to utilize microalgae for precision nutrition and sustainable diets, encompassing the construction of microalgal cell factories, cell cultivation and practical application of microalgae. Manipulating the metabolic pathways and key metabolites of microalgae through multi-omics analysis and employing advanced metabolic engineering strategies, including ZFNs, TALENs, and the CRISPR/Cas system, enhances the production of valuable bioactive compounds, such as omega-3 fatty acids, antioxidants, and essential amino acids. This work begins by providing an overview of the metabolic diversity of microalgae and their ability to thrive in diverse environmental conditions. It then delves into the principles and strategies of metabolic engineering, emphasizing the genetic modifications employed to optimize microalgal strains for enhanced nutritional content. Enhancing PSY, BKT, and CHYB benefits carotenoid synthesis, whereas boosting ACCase, fatty acid desaturases, and elongases promotes polyunsaturated fatty acid production. Here, advancements in synthetic biology, evolutionary biology and machine learning are discussed, offering insights into the precision and efficiency of metabolic pathway manipulation. Also, this review highlights the potential impact of microalgal precision nutrition on human health and aquaculture. The optimized microalgal strains could serve as sustainable and cost-effective sources of nutrition for both human consumption and aquaculture feed, addressing the growing demand for functional foods and environmentally friendly feed alternatives. The tailored microalgal strains are anticipated to play a crucial role in meeting the nutritional needs of diverse populations and contributing to sustainable food production systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Bisphenols (BPs) are recognized as endocrine disrupting compounds and have garnered increasing attention due to their widespread utilization. However, the varying biological toxicities and underlying mechanisms of BPs with different functional groups remain unknown. In the present study, the toxic effects of four BPs (BPA, BPS, BPAF, and TBBPA) on a photosynthetic microalgae Chromochloris zofingiensis were compared. Results showed that halogen-containing BPs exhibited higher cellular uptake, leading to more severe oxidative stress, lower photosynthetic efficiency, and greater accumulation of starch and lipids. Specifically, TBBPA with bromine groups showed a greater toxicity than BPAF with fluorine groups, possibly due to the incomplete debromination in C. zofingiensis. Transcriptomic analysis revealed that halogen-containing BPs triggered greater number of differentially expressed genes (DEGs), and only 64 common DEGs were found among different BPs, indicating that the effects of BPs with different functional groups varied greatly. Genes involved in endocytosis, peroxisomes, and endoplasmic reticulum protein processing pathways were mostly upregulated across different BPs, while photosynthesis-related genes showed varied expression, possibly due to their distinct functional groups. Additionally, SIN3A, ZFP36L, CHMP, and ATF2 emerged as potential key regulatory genes. Overall, this study thoroughly explained how functional groups impact the toxicity and biodegradation of BPs in C. zofingiensis., Competing Interests: Declaration of Competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Microalgae-bacteria consortia (MBC) system has been shown to enhance the efficiency of microalgae in wastewater treatment, yet its effectiveness in treating levofloxacin (LEV) wastewater remains unexplored. This study compared the treatment of LEV wastewater using pure Chlorella pyrenoidosa (PA) and its MBC constructed with activated sludge bacteria. The results showed that MBC improved the removal efficiency of LEV from 3.50-5.41 % to 33.62-57.20 % by enhancing the growth metabolism of microalgae. The MBC increased microalgae biomass and extracellular polymeric substance (EPS) secretion, yet reduced photosynthetic pigment content compared to the PA. At the phylum level, Proteobacteria and Actinobacteriota are the major bacteria in MBC. Furthermore, the transcriptome reveals that the growth-promoting effects of MBC are associated with the up-regulation of genes encoding the glycolysis, the citrate cycle (TCA cycle), and the pentose phosphate pathway. Enhanced carbon fixation, coupled with down-regulation of photosynthetic electron transfer processes, suggests an energy allocation mechanism within MBC. The up-regulation of porphyrin and arachidonic acid metabolism, along with the expression of genes encoding LEV-degrading enzymes, provides evidence of MBC's superior tolerance to and degradation of LEV. Overall, these findings lead to a better understanding of the underlying mechanisms through which MBC outperforms PA in treating LEV wastewater., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Nannochloropsis oceanica is an industrially relevant marine microalga rich in eicosapentaenoic acid (EPA, a valuable ω-3 polyunsaturated fatty acid), yet the algal production potential remains to be unlocked. Here we engineered N. oceanica to synthesize the high-value carotenoid astaxanthin independent of high-light (HL) induction for achieving multifaceted benefits. By screening β-carotenoid ketolases and hydroxylases of various origins, and strategically manipulating compartmentalization, fusion patterns, and linkers of the enzyme pair, a remarkable 133-fold increase in astaxanthin content was achieved in N. oceanica. Iterative metabolic engineering efforts led to further increases in astaxanthin synthesis up to 7.3 mg g -1 , the highest reported for microalgae under nonstress conditions. Astaxanthin was found in the photosystem components and allowed the alga HL resistance and augmented EPA production. Besides, we achieved co-production of astaxanthin and EPA by the engineered alga through a fed-batch cultivation approach. Our findings unveil the untapped potential of N. oceanica as a robust, light-driven chassis for constitutive astaxanthin synthesis and provide feasible strategies for the concurrent production of multiple high-value biochemicals from CO 2 , thereby paving the way for sustainable biotechnological applications of this alga., (© 2024 The Author(s). New Phytologist © 2024 New Phytologist Foundation.)

Microalgae are microorganisms that are rich in bioactive compounds, including pigments, proteins, lipids, and polysaccharides. These compounds can be utilized for a number of biomedical purposes, including drug delivery, wound healing, and tissue engineering. Nevertheless, encapsulating microalgae cells and microalgae bioactive metabolites is vital to protect them and prevent premature degradation. This also enables the development of intelligent controlled release strategies for the bioactive compounds. This review outlines the most employed encapsulation techniques for microalgae, with a particular focus on their biomedical applications. These include ionic gelation, oil-in-water emulsions, and spray drying. Such techniques have been widely explored, due to their ability to protect sensitive compounds from degradation, enhance their stability, extend their shelf life, mask undesirable tastes or odours, control the release of bioactive compounds, and enable targeted delivery to specific sites within the body or environment. Moreover, a patent landscape analysis is also provided, allowing an overview of the microalgae encapsulation technology development applied to a variety of fields, including pharmaceuticals, cosmetics, food, and agriculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Microalgae as a promising approach for wastewater treatment, has challenges in directly treating digested piggery wastewater (DPW) with high ammonia nitrogen (NH 4 + -N) concentration. To improve the performance of microalgae in DPW treatment, straw was employed as a substrate to form a straw-microalgae biofilm. The results demonstrated that the straw-microalgae biofilm achieved the highest NH 4 + -N removal rate of 193.2 mg L -1  d -1 , which was 28.8 % higher than that of culture system without straw. The final NH 4 + -N concentration in the effluent met the discharge standard of 5 mg L -1 . Furthermore, the total organic carbon (TOC) released from straw facilitated bacterial proliferation and the secretion of extracellular polymeric substances (EPS). The EPS and TOC increased the suspension viscosity and surface tension, thereby enhancing the residence time of CO 2 in the liquid phase and promoting CO 2 fixation. This study presented a novel method for the biological treatment of high-ammonia-nitrogen DPW., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Production of functional biocompounds from microalgae has garnered interest from different industrial sectors; however, their overall productivity must be substantially improved for commercialization. Herein, long-term acclimation of Scenedesmus deserticola was conducted using glucose as an organic carbon source to enhance its heterotrophic capabilities and the production potential of loliolide. A year-long acclimation on agar plates led to the selection of S. deserticola HS4, which exhibited at least 2-fold increase in loliolide production potential; S. deserticola HS4 was subjected to further screening of its cultivation conditions and fed-batch cultivation was subsequently performed in liter-scale reactors. While S. deserticola HS4 exhibited shifts in cellular morphology and biochemical composition, the results suggested a substantial increase in its loliolide productivity regardless of trophic modes. Collectively, these results highlight the potential of long-term acclimation as an effective strategy for improving microalgal crops to align with industrial production practices., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Microalgae-based glycolate production through the photorespiratory pathway is considered an environmentally friendly approach. However, the potential for glycolate production is limited by photoautotrophic cultivation with low cell density and existing strains. In this study, a targeted knockout approach was used to disrupt the key photorespiration enzyme, Chlamydomonas reinhardtii hydroxypyruvate reductase 1 (CrHPR1), leading to a significant increase in glycolate production of 280.1 mg/L/OD 750 . The highest potency yield reached 2.1 g/L under optimized mixotrophic conditions, demonstrating the possibility of synchronizing cell growth with glycolate biosynthesis in microalgae. Furthermore, the hypothesis that the cell wall-deficient mutant facilitates glycolate excretion was proposed and validated by comparing the glycolate accumulation trends of various Chlamydomonas reinhardtii strains. This study will facilitate the development of microalgae-based biotechnology and shed lights on the continuous advancement of green biomanufacturing for industrial application., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The study aimed to evaluate the cyanobacteria Spirulina platensis CMB-02 (S. platensis CMB-02) with self-flocculation properties to treat the ammonia nitrogen of rare earth elements (REEs) wastewater. The results demonstrated that S. platensis CMB-02 could effectively remove total ammonia nitrogen (TAN) and total inorganic nitrogen within 5 days. Simultaneously, a self-flocculation efficiency of 82.59 % was achieved by microalga in 30 min after wastewater treatment. The pH, tightly bound extracellular polymeric substances (TB-EPS), and cell morphology of S. platensis CMB-02 were identified as key factors influencing its self-flocculation capabilities. Moreover, the established semi-continuous process with a 20 % renewal rate showed a stable treatment effect, representing a TAN degradation rate of 10.9 mg/(L·d). These obtained findings could conclude that the developed approach mediated with self-flocculating S. platensis CMB-02 was a promising way for REEs wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Ultrasound-assisted extraction (UAE) shows great potential in exploiting microalgal compounds. However, upgrading the extraction system lacks concerns. This study proposes a novel sono-reactor featuring a microbubble distributor for increasing bubble abundance and correspondingly improving microalgal compound extraction. Results indicate that protein concentrations increase with ultrasound powers and extraction time while an optimized gas flow rate exists. The optimal parameters by Box-Behnken design are power 646.0 W, nitrogen flow rate 25.0 mL/min, and time 40.0 min, with an optimal protein concentration of 249.1 mg/L - a substantial improvement over gas-free extraction. The strategic increase in bubble abundance enhances microalgal compound extraction efficiency and extraction kinetics. The system innovation will contribute to the advancement of bioresource utilization and sustainability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Monoclonal antibodies (mAbs) have become indispensable tools in various fields, from research to therapeutics, diagnostics, and industries. However, their production, primarily in mammalian cell culture systems, is cost-intensive and resource-demanding. Microalgae, diverse photosynthetic microorganisms, are gaining attention as a favorable option for manufacturing mAbs and various other recombinant proteins. This review explores the potential of microalgae as a robust expression system for biomanufacturing high-value proteins. It also highlights the diversity of microalgae species suitable for recombinant protein. Nuclear and chloroplast genomes of some microalgae have been engineered to express mAbs and other valuable proteins. Codon optimization, vector construction, and other genetic engineering techniques have significantly improved recombinant protein expression in microalgae. These accomplishments demonstrate the potential of microalgae for biopharmaceutical manufacturing. Microalgal biotechnology holds promise for revolutionizing the production of mAbs and other therapeutic proteins, offering a sustainable and cost-effective solution to address critical healthcare needs., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Microalgal-bacterial granular sludge (MBGS) could offer a sustainable alternative to traditional aerobic methods in brewery wastewater (BWW) treatment. This study compared MBGS with conventional activated sludge (AS) in treating real BWW and highlighted its advantages and challenges. MBGS achieved comparable chemical oxygen demand removal efficiency (93%) compared to AS (89%). Additionally, MBGS exhibited higher phosphate removal capabilities than AS. Extra nitrogen was added to influent to balance C/N ratio of BWW. MBGS was robust in handling C/N ratio fluctuations with an 82% total nitrogen removal efficiency. Metagenomic analysis further indicated that most of the genes involved in carbon, nitrogen and phosphorus metabolism were up-regulated in MBGS compared to AS. Despite changes in the microbial community and settling ability due to high starch and sugar content in BWW, MBGS demonstrated high efficiency and sustainability. Further research should optimize MBGS operation strategies to fully realize its potential for sustainable BWW treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Extracellular soluble algal organic matter (AOM) significantly interferes with microalgae flocculation. This study investigated the effects of various AOM fractions on Chlorella sp. flocculation using ferric chloride (FeCl 3 ), sodium hydroxide (NaOH), and chitosan. All flocculants achieved high separation efficiency (87-99 %), but higher dosages were required in the presence of AOM. High molecular weight (>50 kDa) AOM fraction was identified as the primary inhibitor of flocculation across different pH levels, whereas low/medium molecular weight (<3 and <50 kDa) AOM had minimal impact. Compositional analysis revealed that the inhibitory AOM fraction is a glycoprotein rich in carbohydrates, including neutral, amino, and acidic sugars. The significance of this study is in identifying carboxyl groups (-COOH) from acidic monomers in >50 kDa AOM that inhibit flocculation. Understanding AOM composition and the interaction dynamics between AOM, cells, and flocculants is crucial for enhancing the techno-economics and sustainability of flocculation-based microalgae harvesting., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microalgae have long been regarded as a promising solution for biological carbon abatement from the power industry, offering renewable biomass without competing for land or water resources used for food crops. In this study, we extensively examined the application of photosynthetic microorganisms for closing carbon, nitrogen, and micronutrient loops in the power industry. Subsequently, we explored the bottom-up integration of algal biorefineries into power industry waste streams for increased economic benefits and reduced environmental impacts. Analysis of the available data indicated that microalgae integration with the power industry is primarily performed using flue-gas-assisted cultivation. This approach allows for carbon sequestration typically below one gram per liter per day, too low to significantly impact carbon abatement at achievable scales of microalgae cultivation. Alternative approaches are also being explored. For example, soluble bicarbonate platforms allow for higher biomass productivity and temporary carbon storage. Meanwhile, the use of ashes and waste heat and thermophilic strains can result in lower cultivation costs and better control of cultivation conditions. These approaches offer further incremental improvement to microalgae-based carbon abatement systems in the power industry but are unlikely to be an umbrella solution for carbon reduction. Consequently, in the near term, microalgae-based carbon valorization systems are likely to be limited to niche applications involving the synthesis of high-value products. For microalgae to truly transform carbon abatement processes radical improvements in both biology and engineering approaches are urgently needed.

Microalgae-based biotechnology is one of the most promising alternatives to conventional methods for the removal of antibiotic contaminants from diverse water matrices. However, current knowledge regarding the biochemical mechanisms and catabolic enzymes involved in microalgal biodegradation of antibiotics is scant, which limits the development of enhancement strategies to increase their engineering feasibility. In this study, we investigated the removal dynamics of amphenicols (chloramphenicol, thiamphenicol, and florfenicol), which are widely used in aquaculture, by Chlamydomonas reinhardtii under different growth modes (autotrophy, heterotrophy, and mixotrophy). We found C. reinhardtii removed >92 % chloramphenicol (CLP) in mixotrophic conditions. Intriguingly, gamma-glutamyl hydrolase (GGH) in C. reinhardtii was most significantly upregulated according to the comparative proteomics, and we demonstrated that GGH can directly bind to CLP at the Pro77 site to induce acetylation of the hydroxyl group at C3 position, which generated CLP 3-acetate. This identified role of microalgal GGH is mechanistically distinct from that of animal counterparts. Our results provide a valuable enzyme toolbox for biocatalysis and reveal a new enzymatic function of microalgal GGH. As proof of concept, we also analyzed the occurrence of these three amphenicols and their degradation intermediate worldwide, which showed a frequent distribution of the investigated chemicals at a global scale. This study describes a novel catalytic enzyme to improve the engineering feasibility of microalgae-based biotechnologies. It also raises issues regarding the different microalgal enzymatic transformations of emerging contaminants because these enzymes might function differently from their counterparts in animals., Competing Interests: Declaration of competing interest The authors declared that they have no conflicts of interest in this work. We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microalgae are considered promising sustainable sources of natural bioactive compounds to be used in biotechnological sectors. In recent years, attention is increasingly given to the search of microalgae-derived compounds with antioxidant and anti-inflammatory properties for nutraceutical or pharmacological issues. In this context, attention is usually focused on the composition and bioactivity of algae or their extracts, while less interest is driven to their biological features, for example, those related to morphology and cultivation conditions. In addition, specific studies on the antioxidant and anti-inflammatory properties of microalgae mainly concern Chlorella or Spirulina. The present work was focused on the characterization of the Chlorophyta Neochloris oleoabundans under two combinations of cultivation modes: autotrophy and glucose-induced mixotrophy, each followed by starvation. Biomass for morphological and biochemical characterization, as well as for extract preparation, was harvested at the end of each cultivation phase. Analyses indicated a different content of the most important classes of bioactive compounds with antioxidant/anti-inflammatory properties (lipids, exo-polysaccharides, pigments, total phenolics, and proteins). In particular, the most promising condition able to prompt the production of antioxidant algal biomass with anti-inflammatory properties was the mixotrophic one. Under mixotrophy, beside an elevated algal biomass production, a strong photosynthetic metabolism with high appression of thylakoid membranes and characteristics of high photo-protection from oxidative damage was observed and linked to the overproduction of exo-polysaccharides and lipids rather than pigments. Overall, mixotrophy appears a good choice to produce natural bioactive extracts, potentially well tolerated by human metabolism and environmentally sustainable., (© 2024. The Author(s).)

Algae-derived protein has immense potential to provide high-quality protein foods for the expanding human population. To meet its potential, a broad range of scientific tools are required to identify optimal algal strains from the hundreds of thousands available and identify ideal growing conditions for strains that produce high-quality protein with functional benefits. A research pipeline that includes proteomics can provide a deeper interpretation of microalgal composition and biochemistry in the pursuit of these goals. To date, proteomic investigations have largely focused on pathways that involve lipid production in selected microalgae species. Herein, we report the current state of microalgal proteome measurement and discuss promising approaches for the development of protein-containing food products derived from algae.

Since microalgae face various environmental stresses for the high production of biofuels, multiple studies have been performed to determine if microalgae are resistant to these various stresses. In this study, the viability of cells under various abiotic stresses was investigated by introducing a putative germin-like protein (GLP) from Neopyropia yezoensis , which was known to be related in the resistance to abiotic stresses. The expression of GLP in Chlamydomonas reinhardtii allowed cells to grow better in various abiotic stress environments. In nitrogen starvation conditions, recombinant cells accumulated the lipid droplet 1.46-fold more than wild-type cells and responded more rapidly to form palmelloid forms. Under high-temperature, hydrogen peroxide conditions and saline stress, the survival rate was increased 3.5 times, 2.19 times, and 3.19 times in recombinant C. reinhardtii with GLP, respectively. The expression level of genes related to pathways in response to various stresses increased 2-fold more under those conditions. This result will be useful for the development of microalgae that can grow better and produce more biofuels under different stress conditions.

In recent years, reducing the concentration of carbon dioxide in the atmosphere has become an important issue. Microalgae have a higher photosynthetic efficiency and growth rate than higher plants; thus, biological carbon dioxide fixation using microalgae is attracting particular attention as an efficient carbon dioxide fixation method. However, under dilute atmospheric conditions, microalgae exhibit lower growth rates and reduced carbon dioxide fixation efficiency. In recent years, technology that can efficiently fix carbon dioxide, even in the atmosphere, using a microalgae co-culture system that co-cultivates microalgae and heterotrophic microorganisms has attracted attention. In such a co-culture system, it is believed that a mutualistic relationship is established between microorganisms through the exchange of various compounds. This review focuses on the application of a co-culture system of microalgae and heterotrophic microorganisms for bioproduction and wastewater treatment. In addition, research to elucidate the mutualistic relationships in microalgal co-culture systems using analytical methods that have been widely used in recent years, such as next-generation sequencing technology, is also discussed. In the future, it is expected that the use of microalgae co-culture systems will expand on an industrial scale through the development of key technologies, such as efficient genetic modification techniques for microalgae and their heterotrophic microorganism partners, large-scale cultivation facilities that can efficiently cultivate microalgae, and stable control techniques for co-culture systems using advanced technology., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Microalgae are potential sustainable resources for the production of value-added chemicals that can be used as biofuels, pharmaceuticals, and nutritional supplements. Arachidonic acid (ARA), a omega-6 fatty acid, plays a crucial role in infant development and immune response, and can be used in cosmetics and pharmaceuticals. Demand for industrial-scale ARA production is continuously increasing because of its broad applicability. To address this demand, there has been a significant shift towards microorganism-based ARA production. To accelerate large-scale ARA production, it is crucial to select suitable strains and establish optimal culture conditions., Results: Here, we isolated a novel microalga Lobosphaera incisa CFRC-1, a valuable strain that holds promise as a feedstock for ARA production. Optimal cultivation conditions were investigated using a high-throughput screening method to enhance ARA production in this novel strain. Out of 71 candidates, four organic carbon substrates were identified that could be utilized by L. incisa CFRC-1. Through flask-scale verification, fructose was confirmed as the optimal organic carbon substrate for promoting microalgal growth, total lipid accumulation, and ARA production. Subsequently, we investigated appropriate substrate concentration and cultivation temperature, confirming that the optimal conditions were 30 g L - 1 of fructose and 27 ℃ of temperature. Under these optimized conditions, biomass and ARA production reached 13.05 ± 0.40 g L - 1 and 97.98 ± 7.33 mg L - 1 , respectively, representing 9.6-fold and 5.3-fold increases compared to the conditions before optimization conditions. These results achieved the highest biomass and ARA production in flask-scale cultivation, indicating that our approach effectively improved both production titer and productivity., Conclusions: This study presents a novel microalgae and optimized conditions for enhancing biomass and ARA production, suggesting that this approach is a practical way to accelerate the production of valuable microalgae-based chemicals. These findings provide a basis for large-scale production of ARA-utilizing microalgae for industrial applications., (© 2024. The Author(s).)

The phenolic antioxidant 2,6-Di-tert-butyl-p-cresol (BHT) has been detected in various environments and is considered a potential threat to aquatic organisms. Algal-bacterial interactions are crucial for maintaining ecosystem balance and elemental cycling, but their response to BHT remains to be investigated. This study analyzed the physiological and biochemical responses of the microalga Isochrysis galbana and the changes of associated bacterial communities under different concentrations of BHT stress. Results showed that the biomass of I. galbana exhibited a decreasing trend with increasing BHT concentrations up to 40 mg/L. The reduction in chlorophyll, carotenoid, and soluble protein content of microalgal cells was also observed under BHT stress. The production of malondialdehyde and the activities of superoxide dismutase, peroxidase, and catalase were further determined. Scanning electron microscopy analysis revealed that BHT caused surface rupture of the algal cells and loss of intracellular nutrients. Proteomic analysis demonstrated the upregulation of photosynthesis and citric acid cycle pathways as a response to BHT stress. Additionally, BHT significantly increased the relative abundance of specific bacteria in the phycosphere, including Marivita, Halomonas, Marinobacter, and Alteromonas. Further experiments confirmed that these bacteria had the ability to utilize BHT as the sole carbon resource for growth, and genes related to the degradation of phenolic compounds were detected through pangenome analysis., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

DpAP2 is a transcription factor regulating carotenoid biosynthesis pathway. It was speculated that MeJA significantly decreased expression of DpAP2 gene, then the decreasing DpAP2 expression significantly inhibited expression of some key enzyme genes such as PSY, PDS and GGPS in carotenoid biosynthesis pathway. In contrast, it was speculated that GA3 significantly increased expression of DpAP2 gene, then the increasing DpAP2 expression significantly increased expression of some key enzyme genes such as PDS and GGPS in carotenoid biosynthesis pathway. To increase the content of carotenoid, we evaluated the effect of DpAP2 overexpression on carotenoid accumulation in D. parva. Transgenic D. parva showed a higher carotenoid content (3.18 mg/g DW) compared with control group (2.13 mg/g DW) at 9 d. The dosage effects of exogenous hormones MeJA and GA3 were found in D. parva cells treated with different concentrations of MeJA (10, 20, 50, 100 μM) and GA3 (10, 20, 50, 100 μM). The high concentrations of MeJA (10-100 μM) inhibited the accumulation of carotenoid, and the relative expression of DpAP2, PSY, PDS and GGPS decreased significantly. On the contrary, the relative expression of DpAP2, PDS and GGPS increased significantly when D. parva was treated with 10, 20, 50 and 100 μM GA3, which promoted the biosynthesis of carotenoid. Therefore, we inferred that there was a hierarchical regulation from hormone, transcription factor, key enzyme gene to carotenoid accumulation in carotenoid biosynthesis. Carotenoid biosynthesis was enhanced by DpAP2 overexpression (1.4930 fold of control) and exogenous substances such as GA3 (1.5889 fold of control), which laid a foundation for massive accumulation of carotenoids in microalgae. In the future, further studies were required to demonstrate the complex regulatory network., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ruan et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This study explores the potential of strain selection and adaptation for developing a fungi-yeast-microalgae consortium capable of integrated bioethanol production and livestock wastewater treatment. We employed a multi-stage approach involving isolation and strain selection/adaptation of these consortiums. The study started with screening some isolated fungi to grow on the cellulosic biomass of the livestock wastewater (saccharification) followed by a fermentation process using yeast for bioethanol production. The results revealed that Penicillium chrysogenum (Cla) and Saccharomyces cerevisiae (Sc) produced a remarkable 99.32 ppm of bioethanol and a concentration of glucose measuring 0.56 mg ml - 1 . Following the impact of fungi and yeast, we diluted the livestock wastewater using distilled water and subsequently inoculated Nile River microalgae into the wastewater. The findings demonstrated that Chlorella vulgaris emerged as the dominant species in the microalgal community. Particularly, the growth rate reached its peak at a 5% organic load (0.105385), indicating that this concentration provided the most favorable conditions for the flourishing of microalgae. The results demonstrated the effectiveness of the microalgal treatment in removing the remaining nutrients and organic load, achieving a 92.5% reduction in ammonia, a 94.1% reduction in nitrate, and complete removal of phosphate (100%). The algal treatment also showed remarkable reductions in COD (96.5%) and BOD (96.1%). These findings underscore the potential of fungi, yeast, and Nile River microalgae in the growth and impact on livestock wastewater, with the additional benefit of bioethanol production., (© 2024. The Author(s).)

This study investigated the effects of gamma ( 137 Cs, 0-250 Gy) and UV (UV-C, 0-12 h) radiation on growth and biodiesel properties of Botryococcus braunii KMITL. For gamma radiation, maximum biomass (1.37 ± 0.02 g L -1 ) was achieved with 50 Gy, while a dose of 200 Gy resulted in the highest hydrocarbon content (51.84 ± 0.20%) and yield (0.66 ± 0.01 g L -1 ). For UV radiation, a 9 h exposure produced the highest biomass (2.45 ± 0.05 g L -1 ), hydrocarbon content (55.01 ± 1.22%), and yield (1.35 ± 0.04 g L -1 ). Algae exposed to gamma radiation within the range of 0-150 Gy exhibited C16:0 as the dominant fatty acid methyl ester (FAME), similar to those exposed to UV radiation, while algae exposed to 200-250 Gy displayed C18:1n9t as the dominant FAME. High levels of gamma and UV radiation were observed to lengthen fatty acid chains and increase unsaturated fatty acids. The cetane values of biodiesel from algae exposed to gamma and UV radiation ranged from 64.55 ± 0.14-66.47 ± 0.20 and 59.43 ± 0.04-65.27 ± 0.22, respectively, all meeting standard criteria. Both gamma and UV radiation also improved the saponification value and cold flow properties of the biodiesel. These findings suggest that controlled levels of gamma and UV radiation effectively enhance hydrocarbon yields with significant implications for biofuel production., (© 2024. The Author(s).)

Wastewater generated by the pork industry urgently requires the implementation of low-cost, high-benefit, and efficient treatment systems. Accordingly, a microalgae-bacteria consortia-based treatment system is proposed for the removal of contaminants released, by the pork-producing industry, in swine wastewater. In this study, different inoculum concentrations of the microalgae-bacteria consortium were tested to document variation in the removal of nutrients from the wastewater. At varying concentrations, it was efficient and did not present a significant difference in the removal kinetics. The treatment with the greatest amount of inoculum removed close to 87% of total nitrogen, approximately 70% of orthophosphate, and 77% of chemical oxygen demand. Removals of 84% iron, 44% copper, and 48% manganese were also obtained. These results demonstrate that microalgae-bacteria consortia are an economically viable and environmentally desirable option for the efficient treatment of wastewater from the pork industry., (© 2024. The Author(s).)

Microalgae are significantly influenced by light quality and quantity, whether in their natural habitats or under laboratory and industrial culture conditions. The present study examines the adaptive responses of the marine microalga Tetraselmis chuii to different light regimes, using a cost-effective filtering method and a multi-omics approach. Microalgal growth rates were negatively affected by all filtered light regimes. After six days of cultivation, growth rate for cultures exposed to blue and green filtered light was 67%, while for red filter was 83%, compared to control cultures. Transcriptomic analysis revealed that the usage of green filters resulted in upregulation of transcripts involved in ribosome biogenesis or coding for elongation factors, exemplified by a 2.3-fold increase of TEF3. On the other hand, a 2.7-fold downregulation was observed in photosynthesis-related petJ. Exposure to blue filtered light led to the upregulation of transcripts associated with pyruvate metabolism, while photosynthesis was negatively impacted. In contrast, application of red filter induced minor transcriptomic alterations. Regarding metabolomic analysis, sugars, amino acids, and organic acids exhibited significant changes under different light regimes. For instance, under blue filtered light sucrose accumulated over 6-fold, while aspartic acid content decreased by 4.3-fold. Lipidomics analysis showed significant accumulation of heptadecanoic and linoleic acids under green and red light filters. Together, our findings indicate that filter light can be used for targeted metabolic manipulation., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Aerobic metabolism relies on external oxygen production through photosynthesis and its subsequent transport into each cell of the body via the cardiorespiratory system. This mechanism has successfully evolved over millions of years, enabling animals to inhabit most environments on Earth. However, the insufficient oxygen supply leads to several clinical problems, ranging from non-healing wounds to tumor resistance to therapy. Given that photosynthetic microorganisms are capable of producing oxygen and removing carbon dioxide from the environment, over the last decade, several groups worldwide have proposed their potential use as an alternative tissue oxygenation approach. While most studies have demonstrated safety and efficacy after local tissue administration, recent studies have also suggested that systemic administration could trigger intravascular photosynthesis. If successful, the development of a new generation of circulating cells, known as chlorocytes, may partially replace the role of erythrocytes in gas exchange within the body, without relying on external supply and vascular flow. This work reviews the existing literature on local and systemic administration of photosynthetic microorganisms, highlighting the main challenges in the field and potential solutions to unleash the enormous potential clinical impact of chlorocytes and intravascular photosynthesis. KEY POINTS: • Circulating photosynthetic microorganisms could deliver oxygen to tissues • Microalgae and cyanobacteria have shown safety and efficacy for oxygen delivery • Several key challenges need to be addressed for the clinical success of chlorocytes., (© 2024. The Author(s).)

Microalgae are emerging as a promising feedstock for bioplastics, with Chlorella vulgaris yielding significant amounts of starch. This polysaccharide is convertible into thermoplastic starch (TPS), a biodegradable plastic of industrial relevance. In this study, we developed a pilot-scale protocol for extracting and purifying starch from 430 g (dry weight - DW) of starch-enriched Chlorella vulgaris biomass. More than 200 g DW of starch were recovered, with an extraction yield and starch purity degree reaching 98 and 87 %, respectively. We have characterized this extracted starch and processed it into TPS using twin-screw extrusion and injection molding. Microalgal starch showed similar properties to those of native plant starch, but with smaller granules. We compared the mechanical properties of microalgal TPS with two controls, namely a commercial TPS and a TPS prepared from commercial potato starch granules. TPS prepared from microalgal starch showed a softer and more ductile behavior compared to the reference materials. This study demonstrates the feasibility of recovering high-purity microalgal starch at pilot scale with high yields, and highlights the potential of microalgal starch for the production of TPS using industrially relevant processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Increasing amounts of wastewater is the most pervasive and challenging environmental problem globally. Conventional treatment methods are costly and entail huge energy, carbon consumption and greenhouse gas emissions. Owing to their unique ability of carbon capturing and resource recovery, microalgae-microbiome based treatment is a potential approach and is widely used for carbon-neutral wastewater treatment. Microalgae-bacteria synergy (i.e., the functionally beneficial microbial synthetic communities) performs better and enhances carbon-sequestration and nutrient recovery from wastewater treatment plants. This review presents a comprehensive information regarding the potential of microalgae-microbiome as a sustainable agent for wastewater and discusses synergistic approaches for effective nutrient removal. Moreover, this review discusses, the role of omics-biology and Insilco approaches in unravelling and understanding the algae-microbe synergism and their response toward wastewater treatment. Finally, it discusses various microbiome engineering approaches for developing the effective microalgae-bacteria partners for carbon sequestration and nutrient recovery from wastewater, and summarizes future research perspectives on microalgae-microbiome based bioremediation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Cultivation of microalgae using anaerobic digestate is a gain-win strategy for algal biomass production and achieving environmental benefits. However, the low biomass concentration and high harvest cost of the conventional suspended microalgae culture system are troublesome issues. In this study, a novel fluidized bed photobioreactor (FBPBR) based on diatomite powder was constructed for cultivating Scenedesmus quadricauda and treating diluted anaerobic digestate. The optimized diatomite carrier dosage of 750 mg/L increased microalgal biomass concentration to 1.58 g/L compared to suspended microalgae without carrier (0.99 g/L). When the light intensity was increased from 100 to 200 μmol/m 2 /s, the microalgal biomass in the FBPBR increased to 1.84 g/L and the settling efficiency increased to 93.58 %. This was due to the 1.60-fold enhancement of extracellular polymeric substance (EPS) secretion and changes in EPS properties. The increase in hydrophobic functional groups of EPS under high light intensity, coupled with the reconstitution of protein secondary structure, facilitated the initial attachment of algae to diatomite and the thickening of microalgal biofilm. Moreover, transcriptomic analysis demonstrated that diatomite promoted antioxidant defense and photosynthesis in S. quadricauda cells, alleviating the adverse effect of anaerobic digestate stress. The diatomite addition and elevated light intensity contributed to the highest lipid content (60.37 %), which was owing to the upregulated genes encoding fatty acid and triacylglycerol synthesis under the stress of localized nutrient starvation in the inner layer of microalgae biofilms. Furthermore, the regulation of phosphorus metabolism and NH 4 + -N assimilation improved nutrient removal (93.24 % and 96.86 % for NH 4 + -N and TP removal). This work will provide guidance for the development of FBPBR based on diatomite powder., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Microalgae play an important role in aquatic ecosystems, but the widespread presence of micro- and nano-plastics (MNPs) poses significant threats to them. Haematococcus pluvialis is well-known for its ability to produce the antioxidant astaxanthin when it experiences stress from environmental conditions. Here we examined the effects of polystyrene nanoplastics (PS-NPs) at concentrations of 0.1, 1, and 10 mg/L on H. pluvialis over an 18-day period. Our results show that PS-NPs caused a significant, dose-dependent inhibition of H. pluvialis growth and a reduction in photosynthesis. Furthermore, PS-NPs severely damaged the morphology of H. pluvialis, leading to cell shrinkage, collapse, content release, and aggregation. Additionally, PS-NPs induced a dose-dependent increase in soluble protein content and a decrease in the production of extracellular polymeric substances. These findings indicate that PS-NPs has the potential to adversely affect both the physiology and morphology of H. pluvialis. An increase in reactive oxygen species and antioxidant enzyme activities was also observed, suggesting an oxidative stress response to PS-NPs exposure. Notably, the synthesis of astaxanthin, which is crucial for H. pluvialis's survival under stress, was significantly inhibited in a dose-dependent manner under strong light conditions, along with the down-regulation of genes involved in the astaxanthin biosynthesis pathway. This suggests that PS-NPs exposure reduces H. pluvialis's ability to survive under adverse conditions. This study enhances our understanding of the toxic effects of PS-NPs on microalgae and underscores the urgent need for measures to mitigate MNP pollution to protect aquatic ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Construct a bacteria-algae symbiotic dynamic sponge bioremediation system to simultaneously remove multiple pollutants under micro-pollution conditions. The average removal efficiencies of NH 4 + -N, PO 4 3- -P, total nitrogen (TN), and Ca 2+ were 98.35, 78.74, 95.64, and 84.92 %, respectively. Comparative studies with Auxenochlorella sp. sponge and bacterial sponge bioremediation system confirmed that NH 4 + -N and TN were mainly removed by bacterial heterotrophic nitrification - aerobic denitrification (HN-AD). PO 4 3- -P was removed by algal assimilation and the generation of Ca 3 (PO 4 ) 2 and Ca 5 (PO 4 ) 3 OH, and Ca 2+ was removed by algal electron transfer formation of precipitates and microbially induced calcium precipitation (MICP) by bacteria. Algae provided an aerobic environment for the bacterial HN-AD process through photosynthesis, while respiration produced CO 2 and adsorbed Ca 2+ to promote the formation of calcium precipitates. Immobilization of Ca 2+ with microalgae via bacterial MICP helped to lift microalgal photoinhibition. The bioremediation system provides theoretical support for research on micropolluted water treatment while increasing phosphorus recovery pathways., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Microalgae are susceptible to most pollutants in aquatic ecosystems and can be potentially damaged by silver nanoparticles (AgNPs). This study aims to clarify the potential consequences of Chlorella vulgaris internalizing AgNPs. The exposure of C. vulgaris to AgNPs stabilized with citrate led to the accumulation of NPs in the cell wall, increasing permeability, which allowed the entry of AgNPs and Ag  +  ions resulting from the dissolution of AgNPs. Ag  +  accumulated inside the cell could be converted into AgNPs (endogenous) due to the reducing potential of the cytoplasm. Both exogenous and endogenous AgNPs caused damage to all biological structures of the algae, as demonstrated by TEM images. This damage included the disorganization of chloroplasts, deposition of AgNPs on starch granules, and increased amounts of lipids, starch granules, exopolysaccharides, plastoglobuli, and cell diameters. These changes caused cell death by altering cell viability and interfering with organelle functions, possibly due to reactive oxygen species generated by nanoparticles, as shown in a lipid bilayer model. These findings highlight the importance of considering the exposure risks of AgNPs in a worldwide distributed chlorophyte., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

A critical review and analysis of the literature relevant to the phenolic content of eucaryotic microalgae was performed. Several issues were identified and discussed. In summary, the main problems with the reporting on the phenolic content of microalgae are the following: (1) despite its usefulness in the determination of phenolic content in plant samples, the Folin-Ciocalteu assay is non-suitable for microalgal research due to the high presence of interfering compounds in microalgal extracts such as chlorophyll and its derivatives in organic extracts and free aromatic amino acids or nucleotides in aqueous extracts; (2) while there is chromatographic evidence for the presence of simple phenolic acids in most microalgal clades, the lack of critical enzymes of phenolic biosynthesis in most microalgae, as well as the high variability of phenolic profiles even in the same genus, require more extensive research before conclusions are drawn; (3) the accumulation and metabolism of external phenolics by microalgae has been almost universally neglected in studies focusing on the phenolic content of microalgae, even when natural seawater or complex organic media are used in the cultivation process. Despite these issues, the literature focusing on the bioremediation of waste streams rich in phenolics through microalgae demonstrates the ability of those organisms to adsorb, internalize, and in many cases oxidize or transform a wide range of phenolic compounds, even at very high concentrations. Simple phenolics found in waste streams, such as olive mill waste, have been shown to enhance the antioxidant activity and various bioactivities of microalgal extracts, while complex biotransformation products of phenolics have also been characterized. In conclusion, the de novo biosynthesis of phenolic compounds via eucaryotic microalgae requires further investigation with better designed experiments and suitable analytical methods, while the response of microalgae to phenolic compounds in their growth medium is of great practical interest, both in terms of waste treatment and for the production of functional foods, cosmetics, and pharmaceuticals.

Main Conclusion: PPI analysis deepens our knowledge in critical processes like carbon fixation and nutrient sensing. Moreover, signaling networks, including pathways like MAPK/ERK and TOR, provide valuable information in how microalgae respond to environmental changes and stress. Additionally, species-species interaction networks for microalgae provide a comprehensive understanding of how different species interact within their environments. This review examines recent advancements in the study of biological networks within microalgae, with a focus on the intricate interactions that define these organisms. It emphasizes how network biology, an interdisciplinary field, offers valuable insights into microalgae functions through various methodologies. Crucial approaches, such as protein-protein interaction (PPI) mapping utilizing yeast two-hybrid screening and mass spectrometry, are essential for comprehending cellular processes and optimizing functions, such as photosynthesis and fatty acid biosynthesis. The application of advanced computational methods and information mining has significantly improved PPI analysis, revealing networks involved in critical processes like carbon fixation and nutrient sensing. The review also encompasses transcriptional networks, which play a role in gene regulation and stress responses, as well as metabolic networks represented by genome-scale metabolic models (GEMs), which aid in strain optimization and the prediction of metabolic outcomes. Furthermore, signaling networks, including pathways like MAPK/ERK and TOR, are crucial for understanding how microalgae respond to environmental changes and stress. Additionally, species-species interaction networks for microalgae provide a comprehensive understanding of how different species interact within their environments. The integration of these network biology approaches has deepened our understanding of microalgal interactions, paving the way for more efficient cultivation and new industrial applications., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Co-culturing fungi and microalgae may effectively remediate wastewater containing Cd and harvest microalgae. Nevertheless, a detailed study of the mechanisms underlying the synergistic interactions between fungi and microalgae under Cd(II) exposure is lacking. In this study, Cd(II) exposure resulted in a significant enhancement of antioxidants, such as glutathione (GSH), malondialdehyde (MDA), hydrogen peroxide (H 2 O 2 ) and superoxide dismutase (SOD) compared to the control group, suggesting that the cellular antioxidant defense response was activated. Extracellular proteins and extracellular polysaccharides of the symbiotic system were increased by 60.61 % and ,24.29 %, respectively, after Cd(II) exposure for 72 h. The adsorption behavior of Cd(II) was investigated using three-dimensional fluorescence excitation-emission matrix (3D-EEM), fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM). Metabolomics results showed that the TCA cycle provided effective material and energy supply for the symbiotic system to resist the toxicity of Cd(II); Proline, histidine, and glutamine strengthened the synergistic adsorption capacity of the fungus and microalgae. Overall, the theoretical foundation for a deep comprehension of the beneficial interactions between fungi and microalgae under Cd(II) exposure and the role of the fungal-algal symbiotic system in the management of heavy metal pollution is provided by this combined physiological and metabolomic investigation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The neurotoxin β-N-methylamino-L-alanine (BMAA), produced by cyanobacteria and diatoms, has been implicated as an environmental risk factor for neurodegenerative diseases. This study first investigated the occurrence and monthly distributions of BMAA and its isomers, 2,4-diaminobutyric acid (DAB) and N-2-aminoethylglycine (AEG), in phytoplankton and mussels from 11 sites along the South Sea Coast of Korea throughout 2021. These toxins were quantified using LC-MS/MS, revealing elevated BMAA concentrations from late autumn to spring, with phase lags observed between phytoplankton and mussels. The highest concentration of BMAA in phytoplankton was detected in November (mean: 1490 ng g -1 dry weight (dw)), while in mussels, it peaked in December (mean: 1240 ng g -1 dw). DAB was detected in phytoplankton but was absent in mussels, indicating limited bioaccumulation potential. In February, the peak mean DAB concentration in phytoplankton was 89 ng g -1 dw. AEG was not detected in any samples. Chlorophyll-a concentrations consistently showed an inverse correlation with BMAA concentrations in mussels throughout the year. Through correlation analysis, four diatom genera, Bacillaria, Hemiaulus, Odontella, and Pleurosigma, were identified as potential causative microalgae of BMAA. This study offers insights into identifying the causative microalgae for BMAA and informs future regulatory efforts regarding unmanaged biotoxins., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Effectively treating phenolic substances is a crucial task in environmental protection. This study aims to determine whether bacterial-algae biofilm reactors offer superior treatment efficacy compared to traditional activated sludge and biofilm reactors. The average degradation ratios of 2,4-dimethylphenol (40, 70, 150, 300, and 230 mg/L) were found to be 98 %, 99 %, 92.1 %, 84.7 %, and 63.7 % respectively. The bacterial-algae biofilm demonstrates a higher tolerance to toxicity, assimilation ability, and efficacy recovery ability. The cell membrane of Chlorella in the bacteria-algae biofilm is not easily compromised, thus ensuring a stable pH environment. High concentrations of tightly bound extracellular polymers (TB-EPS) enhance the efficacy in treating toxic pollutants, promote the stable structure. Intact Chlorella, bacilli, and EPS were observed in bacterial-algal biofilm. The structural integrity of bacteria-algae consistently enhances its resistance to the inhibitory effects of high concentrations of phenolic compounds. Cloacibacterium, Comamonas, and Dyella were the main functional bacterial genera that facilitate the formation of bacterial-algal biofilms and the degradation of phenolic compounds. The dominant microalgal families include Aspergillaceae, Chlorellales, Chlorellaceae, and Scenedesmaceae have certain treatment effects on phenolic substances. Chlorellales and Chlorellaceae have the ability to convert NH 4 + -N. The Aspergillaceae is also capable of generating synergistic effects with Chlorellales, Chlorellaceae, and Scenedesmaceae, thereby establishing a stable bacterial-algal biofilm system., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Biocomponents (such as lipids) accumulate in oleaginous microorganisms and could be used for renewable energy production. Oleaginous microbes are characterized by their ability to accumulate high levels of lipids, which can be converted into biodiesel. The oleaginous microbes (including microalgae, bacteria, yeast, and fungi) can utilize diverse substrates. Thus, in this study, commercially viable oleaginous microorganisms are comparatively summarized for their growth conditions, substrate utilization, and applications in biotechnological processes. Lipid content is species-dependent, as are culture conditions (such as temperature, pH, nutrients, and culture time) and substrates. Lipid production can be increased by selecting suitable microorganisms and substrates, optimizing environmental conditions, and using genetic engineering techniques. In addition, the emphasis on downstream processes (including harvesting, cell disruption, lipid extraction, and transesterification) highlights their critical role in enhancing cost-effectiveness. Oleaginous microorganisms are potential candidates for lipid biosynthesis and could play a key role in meeting the energy needs of the world in the future., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Aim: Ammonium chloride (NH4Cl) is one of the nitrogen sources for microalgal cultivation. An excessive amounts of NH4Cl are toxic for microalgae. However, combining mixotrophic conditions and excessive quantities of NH4Cl positively affects microalgal biomass and lipid production. In this study, we investigated the impact of NH4Cl on the growth, biomass, and triglyceride (TAG) content of the green microalga Chlamydomonas reinhardtii especially under mixotrophic conditions., Methods and Results: Under photoautotrophic conditions (without organic carbon supplementation), adding 25 mM NH4Cl had no significant effect on microalgal growth or TAG content. However, under mixotrophic condition (with acetate supplementation), NH4Cl interfered with microalgal growth while inducing TAG content. To explore these effects further, we conducted a two-step cultivation process and found that NH4Cl reduced microalgal growth, but induced total lipid and TAG content, especially after 4-day cultivation. The photosynthesis performances showed that NH4Cl completely inhibited oxygen evolution on day 4. However, NH4Cl slightly reduced the Fv/Fm ratio indicating that the NH4Cl supplementation directly affects microalgal photosynthesis. To investigate the TAG induction effect by NH4Cl, we compared the protein expression profiles of microalgae grown mixotrophically with and without 25 mM NH4Cl using a proteomics approach. This analysis identified 1782 proteins, with putative acetate uptake transporter GFY5 and acyl-coenzyme A oxidase being overexpressed in the NH4Cl-treated group., Conclusion: These findings suggested that NH4Cl supplementation may stimulate acetate utilization and fatty acid synthesis pathways in microalgae cells. Our study indicated that NH4Cl supplementation can induce microalgal biomass and lipid production, particularly when combined with mixotrophic conditions., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

This study investigated the impact of culture medium salinity (5-50 PSU) on the growth and maximum photochemical yield of photosystem II ( Fv / Fm ) and the composition of carotenoids, fatty acids, and bioactive substances in three marine microalgae ( Chrysochromulina rotalis , Amphidinium carterae , and Heterosigma akashiwo ). The microalgae were photoautotrophically cultured in discontinuous mode in a single stage (S1) and a two-stage culture with salt shock (S2). A growth model was developed to link biomass productivity with salinity for each species. C. rotalis achieved a maximum biomass productivity ( Pmax ) of 15.85 ± 0.32 mg·L -1 ·day -1 in S1 and 16.12 ± 0.13 mg·L -1 ·day -1 in S2. The salt shock in S2 notably enhanced carotenoid production, particularly in C. rotalis and H. akashiwo , where fucoxanthin was the main carotenoid, while peridinin dominated in A. carterae . H. akashiwo also exhibited increased fatty acid productivity in S2. Salinity changes affected the proportions of saturated, monounsaturated, and polyunsaturated fatty acids in all three species. Additionally, hyposaline conditions boosted the production of haemolytic substances in A. carterae and C. rotalis.

Microalgal solutions to clean waste streams and produce biomass were evaluated in Nordic conditions during winter, spring, and autumn in Southeast Sweden. The study investigated nitrogen (N) removal, biomass quality, and safety by treating industrial leachate water with a polyculture of local microalgae and bacteria in open raceway ponds, supplied with industrial CO 2 effluent. Total N (TN) removal was higher in spring (1.5 g -2 d -1 ), due to beneficial light conditions compared to winter and autumn (0.1 and 0.09 g -2 d -1 ). Light, TN, and N species influenced the microalgal community (dominated by Chlorophyta), while the bacterial community remained stable throughout seasons with a large proportion of cyanobacteria. Winter conditions promoted biomass protein (19.6-26.7%) whereas lipids and carbohydrates were highest during spring (11.4-18.4 and 15.4-19.8%). Biomass toxin and metal content were below safety levels for fodder, but due to the potential presence of toxic strains, biofuels or fertilizer could be suitable applications for the algal biomass. PRACTITIONER POINTS: Microalgal removal of nitrogen from leachate water was evaluated in Nordic conditions during winter, spring, and autumn. Total nitrogen removal was highest in spring (1.5 g -2 d -1 ), due to beneficial light conditions for autotrophic growth. Use of local polyculture made the cultivation more stable on a seasonal (light) and short-term (N-species changes) scale. Toxic elements in produced algal biomass were below legal thresholds for upcycling., (© 2024 The Author(s). Water Environment Research published by Wiley Periodicals LLC on behalf of Water Environment Federation.)

β-N-Methylamino-L-alanine (BMAA), a neurotoxin produced by various microalgal groups, is associated with neurodegenerative diseases and is considered a major environmental factor potentially linked to sporadic amyotrophic lateral sclerosis. This study systematically reviews the analytical methods used to study BMAA in publications from 2019 to the present. It also investigates the causative microalgae of BMAA and its geographical distributions in aquatic ecosystems based on studies conducted since 2003. A comprehensive search using the Web of Science database revealed that hydrolysis for extraction (67%), followed by quantification using LC-MS/MS (LC: 84%; MS/MS: 88%), is the most commonly employed method in BMAA analysis. Among analytical methods, RPLC-MS/MS had the highest percentage (88%) of BMAA-positive results and included a high number of quality control (QC) assessments. Various genera of cyanobacteria and diatoms have been reported to produce BMAA. The widespread geographical distribution of BMAA across diverse ecosystems highlights significant environmental and public health concerns. Notably, BMAA accumulation and biomagnification are likely more potent in marine or brackish water ecosystems than in freshwater ecosystems, potentially amplifying its ecological impacts. Future research should prioritize advanced, sensitive methods, particularly LC-MS/MS with as many QC assessments as possible, and should expand investigations to identify novel microalgal producers and previously uncharted geographical areas, with a special focus on marine or brackish water ecosystems. This effort will enhance our understanding of the environmental distribution and impacts of BMAA., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The aquaculture industry is under the framework of the food-water-energy nexus due to the extensive use of water and energy. Sustainable practices are required to support the tremendous growth of this sector. Currently, the aquaculture industry is challenged by its reliance on capture fisheries for feed, increased use of pharmaceuticals, infectious outbreaks, and solid/liquid waste management. This review posits microalgal technology as a comprehensive solution for the current predicaments in aquaculture in a sustainable way. Microalgae are microscopic, freshwater and marine photosynthetic organisms, capable of carbon mitigation and bioremediation. They are indispensable in aquaculture due to their key role in marine productivity and their position in the marine food chain. Microalgae are nutritious and are currently used as feed in specific sectors of aquaculture. Due to their bioremediation potential, direct application of microalgae in shellfish ponds and in recirculating systems have been adopted to improve water quality and aquatic animal health. The potential of microalgae for integration into various aspects of aquaculture processes, namely hatcheries, feed, and waste management has been critically analyzed. Seamless integration of microalgal technology in aquaculture is feasible, and this review will provide new insights into using microalgal technology for sustainable aquaculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Coal mining endangers the environment by contaminating of soil, surface, and ground water with coal mine drainage water (CMW) polluted by heavy metals. Microalgal cultures, hyper-accumulators of heavy metals, represent a promising solution for CMW biotreatment. A bottleneck of this approach is the availability of microalgal strains that combine a large capacity for heavy metal biocapture with a high resilience to their toxic effects. Biotopes contaminated with heavy metals are frequently inhabited by microalgae evolved to be resilient to heavy metal toxicity. Therefore, the autochthonous (locally isolated) microalgal strains are a priori considered to be superior for biotreatment of heavy metal-polluted waste streams. Still, strains from biocollections combine a high pollutant resilience with other biotechnologically important traits such as high productivity, high CO 2 sequestration rate etc. Moreover, the strains available "off-the-shelf" would enable rapid development of bioprocesses. Here, we compared the efficiency of CMW biotreatment with autochthonous (isolated from the coal mine drainage sump) and allochthonous microalgae (from a geographically distant phosphate-polluted site). Both autochthonous strains and allochthonous strains turned to be interchangeable under our experimental conditions. Still, the autochthonous strains showed a higher capacity for sequestration of iron, zinc, and manganese, the specific pollutants of the studied CMW. It can be important when the duration of unattended exploitation of the CMW treatment facility is a priority or spikes of the heavy metal concentration in CMW are expected. Therefore, the "off-the-shelf" strains can be a plausible solution for rapid development of CMW treatment technologies from scratch (although screening for acute toxicity of CMW is imperative). On the other hand, locally isolated strains can offer distinct advantages and should be always considered if sufficient time and other resources are available for the development of microalgae-based process for CMW treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Food processing wastewater presents a considerable challenge for treatment owing to its elevated nitrogen and phosphorus levels. Nonetheless, it possesses inherent value attributed to its abundant nutrients and organic content. This study presents an innovative approach for treating food processing wastewater and reusing biomass. Initially, the secondary-treated wastewater undergoes flocculation and sedimentation, followed by reverse osmosis to ensure that the effluent meets reuse standards. Subsequently, reverse osmosis concentrates, generated at varying water recovery rates, are utilized for microalgae cultivation to recover nitrogen and phosphorus. Furthermore, this study highlights the potential of reverse osmosis concentrates in reducing the water demand for microalgae cultivation and in producing commercial-grade nutrients. The findings reveal that reverse osmosis achieves removal rates exceeding 90 % for both nitrogen and phosphorus and effluent meets reuse standards. Following seven days of cultivation, microalgae cultured in reverse osmosis concentrated water with an 80 % water recovery rate demonstrate denitrification and phosphorus removal rates of 73.88 % and 80.92 % respectively, with a biomass concentration of 563 mg/L and a protein yield of 128 mg/L. Moreover, a total volumetric energy yield of 10.08 kJ/L is obtained, facilitating energy valorization. In conclusion, this study offers practical solutions for wastewater treatment and resource recovery, enabling the attainment of zero discharge of pollutants while generating valuable resources through microalgae cultivation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)