Influence of salinity, nitrogen and phosphorus concentrations on the physiological and biochemical characteristics of two Chlorophyceae isolated from Fez freshwater, Morocco.

Microalgae are widely exploited for numerous biotechnology applications, including biofuels. In this context, Chlamydomonas debaryana and Chlorococcum sp. were isolated from Fez freshwater (Morocco), and their growth and lipid and carbohydrate production were assessed at different concentrations of NaCl, NaNO3, and K2HPO4. The results indicate a small positive variation in growth parameters linked to nutrient enrichment, with no considerable variation in carbohydrate and lipid levels in both algae. Moreover, a negative variation was recorded at increased salinity and nutrient limitation, accompanied by lipid and carbohydrate accumulation. Chlorococcum sp. showed better adaptation to salt stress below 200 mM NaCl. Furthermore, its growth and biomass productivity were strongly reduced by nitrogen depletion, and its lipid production reached 47.64% DW at 3.52 mM NaNO3. As for Chlamydomonas debaryana, a substantial reduction in growth was induced by nutrient depletion, a maximal carbohydrate level was produced at less than 8.82 mM NaNO3 (40.59% DW). The effect of phosphorus was less significant. However, a concentration of 0.115 mM K2HPO4 increased lipid and carbohydrate content without compromising biomass productivity. The results suggest that growing the two Chlorophyceae under these conditions seems interesting for biofuel production, but the loss of biomass requires a more efficient strategy to maximize lipid and carbohydrate accumulation without loss of productivity.

Nutrients recovery from dairy wastewater by Chlorella vulgaris and comparison of the lipid's composition with various chlorella strains for biodiesel production.

Microalgae biomass is regarded as a promising feedstock for biodiesel production. The biomass lipid content and fatty acids composition are among the main selective criteria when screening microalgae strains for biodiesel production. In this study, three strains of Chlorella microalgae (C. kessleri, C. sorokiniana, C. vulgaris) were cultivated nutrient media with different nitrogen contents, and on a medium with the addition of dairy wastewater. Moreover, microalgae grown on dairy wastewater allowed the removal of azote and phosphorous. The removal efficiency of 90%, 53% and 95% of ammonium nitrogen, total nitrogen and phosphate ions, respectively, were reached. The efficiency of wastewater treatment from inorganic carbon was 55%, while the maximum growth of biomass was achieved. All four samples of microalgae had a similar fatty acid profile. Palmitic acid (C16:0) was the most abundant saturated fatty acid (SFA), and is suitable for the production of biodiesel. The main unsaturated fatty acids (UFA) present in the samples were oleic acid (C18:1 n9); linoleic acid (C18:2 n6) and alpha-linolenic acid (C18:3 n3), which belong to omega-9, omega-6, omega-3, respectively.

Harnessing the potential of microalgae-based systems for mitigating pesticide pollution and its impact on their metabolism.

Due to increased pesticide usage in agriculture, a significant concentration of pesticides is reported in the environment that can directly impact humans, aquatic flora, and fauna. Utilizing microalgae-based systems for pesticide removal is becoming more popular because of their environmentally friendly nature, ability to degrade pesticide molecules into simpler, nontoxic molecules, and cost-effectiveness of the technology. Thus, this review focused on the efficiency, mechanisms, and factors governing pesticide removal using microalgae-based systems and their effect on microalgal metabolism. A wide range of pesticides, like atrazine, cypermethrin, malathion, trichlorfon, thiacloprid, etc., can be effectively removed by different microalgal strains. Some species of Chlorella, Chlamydomonas, Scenedesmus, Nostoc, etc., are documented for >90% removal of different pesticides, mainly through the biodegradation mechanism. The antioxidant enzymes such as ascorbate peroxidase, superoxide dismutase, and catalase, as well as the complex structure of microalgae cell walls, are mainly involved in eliminating pesticides and are also crucial for the defense mechanism of microalgae against reactive oxygen species. However, higher pesticide concentrations may alter the biochemical composition and gene expression associated with microalgal growth and metabolism, which may vary depending on the type of strain, the pesticide type, and the concentration. The final section of this review discussed the challenges and prospects of how microalgae can become a successful tool to remediate pesticides.

Microalgae to biodiesel: A novel green conversion method for high-quality lipids recovery and in-situ transesterification to fatty acid methyl esters.

Greenhouse gases (GHGs) emissions due to increasing energy demand have raised the need to identify effective solutions to produce clean and renewable energy. Biotechnologies are an effective platform to attain green transition objectives, especially when synergically integrated to promote health and environmental protection. In this context, microalgae-based biotechnologies are considered among the most effective tools for treating gaseous effluents and simultaneously capturing carbon sources for further biomass valorisation. The production of biodiesel is regarded as a promising avenue for harnessing value from residual algal biomass. Nonetheless, the existing techniques for extracting lipids still face certain limitations, primarily centred around the cost-effectiveness of the process.This study is dedicated to developing and optimising an innovative and cost-efficient technique for extracting lipids from algal biomass produced during gaseous emissions treatment based on algal-bacterial biotechnology. This integrated treatment technology combines a bio-scrubber for degrading gaseous contaminants and a photobioreactor for capturing the produced CO2 within valuable algal biomass. The cultivated biomass is then processed with the process newly designed to extract lipids simultaneously transesterificated in fatty acid methyl esters (FAME) via In Situ Transesterification (IST) with a Kumagawa-type extractor. The results of this study demonstrated the potential application of the optimised method to overcome the gap to green transition. Energy production was obtained from residuals produced during the necessary treatment of gaseous emissions. Using hexane-methanol (v/v = 19:1) mixture in the presence KOH in Kumagawa extractor lipids were extracted with extraction yield higher than 12% and converted in fatty acid methyl esters. The process showed the enhanced extraction of lipids converted in bio-sourced fuels with circular economy approach, broadening the applicability of biotechnologies as sustainable tools for energy source diversification.

Tetraselmis chuii Edible Microalga as a New Source of Neuroprotective Compounds Obtained Using Fast Biosolvent Extraction.

Tetraselmis chuii is an EFSA-approved novel food and dietary supplement with increasing use in nutraceutical production worldwide. This study investigated the neuroprotective potential of bioactive compounds extracted from T. chuii using green biobased solvents (ethyl acetate, AcOEt, and cyclopentyl methyl ether, CPME) under pressurized liquid extraction (PLE) conditions and supercritical fluid extraction (SFE). Response surface optimization was used to study the effect of temperature and solvent composition on the neuroprotective properties of the PLE extracts, including anticholinergic activity, reactive oxygen/nitrogen species (ROS/RNS) scavenging capacity, and anti-inflammatory activity. Optimized extraction conditions of 40 °C and 34.9% AcOEt in CPME resulted in extracts with high anticholinergic and ROS/RNS scavenging capacity, while operation at 180 °C and 54.1% AcOEt in CPME yielded extracts with potent anti-inflammatory properties using only 20 min. Chemical characterization revealed the presence of carotenoids (neoxanthin, violaxanthin, zeaxanthin, α- and ß-carotene) known for their anti-cholinesterase, antioxidant, and anti-inflammatory potential. The extracts also exhibited high levels of omega-3 polyunsaturated fatty acids (PUFAs) with a favorable ω-3/ω-6 ratio (>7), contributing to their neuroprotective and anti-inflammatory effects. Furthermore, the extracts were found to be safe to use, as cytotoxicity assays showed no observed toxicity in HK-2 and THP-1 cell lines at or below a concentration of 40 µg mL-1. These results highlight the neuroprotective potential of Tetraselmis chuii extracts, making them valuable in the field of nutraceutical production and emphasize the interest of studying new green solvents as alternatives to conventional toxic solvents.

Effects of Supplementation with Microalgae Extract from Phaeodactylum tricornutum (Mi136) to Support Benefits from a Weight Management Intervention in Overweight Women.

BACKGROUND: Microalgae like Phaeodactylum tricornutum (PT) contain the carotenoid, fucoxanthin, which has been purported to promote fat loss, lower blood lipids, and improve glucose management. This study examined whether dietary supplementation with microalgae extracts from PT containing 4.4 mg/d of fucoxanthin affects changes in body composition or health markers in overweight women during an exercise and diet intervention. MATERIALS AND METHODS: A total of 37 females (28.6 ± 7.9 years, 80.2 ± 14.9 kg, 29.6 ± 3.8 kg/m², 41.4 ± 4.2% fat) fasted for 12 h, donated a fasting blood sample, completed health and mood state inventories, and undertook body composition, health, and exercise assessments. In a counterbalanced, randomized, and double-blind manner, participants ingested a placebo (PL), or microalgae extract of Phaeodactylum tricornutum standardized to 4.4 mg of fucoxanthin (FX) for 12 weeks while participating in a supervised exercise program that included resistance-training and walking (3 days/week) with encouragement to accumulate 10,000 steps/day on remaining days of the week. The diet intervention involved reducing energy intake by about -300 kcal/d (i.e., ≈1400-1600 kcals/d, 55% carbohydrate, 30% fat, 15% protein) to promote a -500 kcal/d energy deficit with exercise. Follow-up testing was performed at 6 and 12 weeks. A general linear model (GLM) with repeated measures statistical analysis was used to analyze group responses and changes from baseline with 95% confidence intervals. RESULTS: Dietary supplementation with microalgae extract from PT containing fucoxanthin for 12 weeks did not promote additional weight loss or fat loss in overweight but otherwise healthy females initiating an exercise and diet intervention designed to promote modest weight loss. However, fucoxanthin supplementation preserved bone mass, increased bone density, and saw greater improvements in walking steps/day, resting heart rate, aerobic capacity, blood lipid profiles, adherence to diet goals, functional activity tolerance, and measures of quality of life. Consequently, there appears to be some benefit to supplementing microalgae extract from PT containing fucoxanthin during a diet and exercise program. Registered clinical trial #NCT04761406.

In Vivo Absorption and Lymphatic Bioavailability of Docosahexaenoic Acid from Microalgal Oil According to Its Physical and Chemical Form of Vectorization.

Docosahexaenoic acid (DHA) is an essential fatty acid (FA) with proven pro-health effects, but improving its bioavailability is becoming a public health issue. The bioavailability of DHA from microalgal (A) oil has been comprehensively assessed, particularly in terms of the molecular structuring capabilities offered by A-oil. Here, we explored the impact of five DHA-rich formulas differing in terms of (i) molecular structure, i.e., ethyl ester (EE), monoglyceride (MG), or triglyceride (TG), and (ii) supramolecular form, i.e., emulsified TG or TG + phospholipids (PL blend) on the lymphatic kinetics of DHA absorption and the lipid characteristics of the resulting lipoproteins. We demonstrated in rats that the conventional A-DHA TG structure afforded more effective DHA absorption than the EE structure (+23%). Furthermore, the A-DHA MG and A-DHA emulsions were the better DHA vectors (AUC: 89% and +42%, respectively) due to improved lipolysis. The A-DHA MG and A-DHA emulsion presented the richest DHA content in TG (+40%) and PL (+50%) of lymphatic chylomicrons, which could affect the metabolic fate of DHA. We concluded that structuring A-DHA in TG or EE form would better serve for tissue and hepatic metabolism whereas A-DHA in MG and emulsion form could better target nerve tissues.

Microalgae enhanced co-metabolism of sulfamethoxazole using aquacultural feedstuff components: Co-metabolic pathways and enzymatic mechanisms.

The application of antibiotics in freshwater aquaculture leads to increased contamination of aquatic environments. However, limited information is available on the co-metabolic biodegradation of antibiotics by microalgae in aquaculture. Feedstuffs provide multiple organic substrates for microalgae-mediated co-metabolism. Herein, we investigated the co-metabolism of sulfamethoxazole (SMX) by Chlorella pyrenoidosa when adding main components of feedstuff (glucose and lysine). Results showed that lysine had an approximately 1.5-fold stronger enhancement on microalgae-mediated co-metabolism of SMX than glucose, with the highest removal rate (68.77% ± 0.50%) observed in the 9-mM-Lys co-metabolic system. Furthermore, we incorporated reactive sites predicted by density functional theory calculations, 14 co-metabolites identified by mass spectrometry, and the roles of 18 significantly activated enzymes to reveal the catalytic reaction mechanisms underlying the microalgae-mediated co-metabolism of SMX. In lysine- and glucose-treated groups, five similar co-metabolic pathways were proposed, including bond breaking on the nucleophilic sulfur atom, ring cleavage and hydroxylation at multiple free radical reaction sites, together with acylation and glutamyl conjugation on electrophilic nitrogen atoms. Cytochrome P450, serine hydrolase, and peroxidase play crucial roles in catalyzing hydroxylation, bond breaking, and ring cleavage of SMX. These findings provide theoretical support for better utilization of microalgae-driven co-metabolism to reduce sulfonamide antibiotic residues in aquaculture.

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.

Simultaneous exposure to nanoplastics and cadmium mitigates microalgae cellular toxicity: Insights from molecular simulation and metabolomics.

In the severe pollution area of nanoplastics (NPs) and cadmium ions (Cd2+), the joint effects of their high environmental concentrations on primary producers may differ from those of low environmental doses. Thus, we investigated the physiological changes, cell morphology, molecular dynamic simulation, phenotypic interactions, and metabolomics responses of C. pyrenoidosa to high environmental concentrations of NPs and Cd2+ after 12-d acclimation. After 12-d cultivation, mono-NPs and mono-Cd2+ reduced cell density and triggered antioxidant enzymes, extracellular polymeric substances (EPS) production, and cell aggregation to defend their unfavorable effects. Based on the molecular dynamic simulation, the chlorine atoms of the NPs and Cd2+ had charge attraction with the nitrogen and phosphorus atoms in the choline and phosphate groups in the cell membrane, thereby NPs and Cd2+ could adsorb on the cells to destroy them. In the joint exposure, NPs dominated the variations of ultrastructure and metabolomics and alleviated the toxicity of NPs and Cd2+. Due to its high environmental concentration, more NPs could compete with the microalgae for Cd2+ and thicken cell walls, diminishing the Cd2+ content and antioxidant enzymes of microalgae. NPs addition also decreased the EPS content, while the bound EPS with -CN bond was kept to detoxicate Cd2+. Metabolomics results showed that the NPs downregulated nucleotide, arachidonic acid, and tryptophan metabolisms, while the Cd2+ showed an opposite trend. Compared with their respective exposures, metabolomics results found the changes in metabolic molecules, suggesting the NPs_Cd2+ toxicity was mitigated by balancing nucleotide, arachidonic acid, tryptophan, and arginine and proline metabolisms. Consequently, this study provided new insights that simultaneous exposure to high environmental concentrations of NPs and Cd2+ mitigated microalgae cellular toxicity, which may change their fates and biogeochemical cycles in aquatic systems.

Microalgae as future food: Rich nutrients, safety, production costs and environmental effects.

The improvement of food security and nutrition has attracted wide attention, and microalgae as the most promising food source are being further explored. This paper comprehensively introduces basic and functional nutrients rich in microalgae by elaborated tables incorporating a wide variety of studies and summarizes factors influencing their accumulation effects. Subsequently, multiple comparisons of nutrients were conducted, indicating that microalgae have a high protein content. Moreover, controllable production costs and environmental friendliness prompt microalgae into the list that contains more promising and reliable future food. However, microalgae and -based foods approved and sold are limited strictly, showing that safety is a key factor affecting dietary consideration. Notably, sensory profiles and ingredient clarity play an important role in improving the acceptance of microalgae-based foods. Finally, based on the bottleneck in the microalgae food industry, suggestions for its future development were discussed.

A review of emerging membrane-based microalgal-bacterial processes for wastewater treatment: Process configurations, biological and membrane performance, and perspectives.

Microalgal-bacterial (MB) consortia create an excellent eco-system for simultaneous COD/BOD and nutrients (N and P) removals in a single step with significant reduction in or complete elimination of aeration and carbonation in the biological wastewater treatment processes. The integration of membrane separation technology with the MB processes has created a new paradigm for research and development. This paper focuses on a comprehensive and critical literature review of recent advances in these emerging processes. Novel membrane process configurations and process conditions affecting the biological performance of these novel systems have been systematically reviewed and discussed. Membrane fouling issues and control of MB biofilm formation and thickness associated with these emerging suspended growth or immobilized biofilm processes are addressed and discussed. The research gaps, challenges, outlooks of these emerging processes are identified and discussed in-depth. The findings from the literature suggest that the membrane-based MB processes are advanced biotechnologies with a significant reduction in energy consumption and process simplification and high process efficiency that are not achievable with current technologies in wastewater treatment. There are endless opportunities for research and development of these novel and emerging membrane-based MB processes.

Injectable Dynamic ROS-Responsive COF-Modified Microalgae Gels for In Vivo bFGF Delivery to Treat Diabetic Wounds.

Hypoxia, chronic inflammation, and elevated reactive oxygen species (ROS) production induced by hyperglycemia pose formidable challenges to the healing of diabetic chronic wounds, often resulting in impaired recovery. Currently, sustainable and eco-friendly therapeutic approaches targeting this multifaceted problem remain uncharted. Herein, we develop a unique three-functional covalent organic framework (COF)-modified microalgae gel designed for the preparation and treatment of chronic diabetic wounds. The gel comprises an oxygen-releasing basic fibroblast growth factor (bFGF) microalgae matrix, augmented by an ROS-responsive COF. Although two of these components have been reported to be used in wound healing, the combination of all three functions represents an innovative approach to synergize the treatment of chronic diabetic wounds. Therefore, we propose a new concept of "ligand interlocking" with three functional synergistic effects. Specifically, the COF has a similar effect to the "double Excalibur", which binds bFGF to promote angiogenesis and proliferation and inhibit the inflammatory response of chronic wounds and binds live microalgae to eliminate ROS and release dissolved oxygen to alleviate the hypoxia of wounds. Moreover, in vivo experiments and RNA sequencing analyses similarly demonstrated that the COF-modified microalgae gel reduced the inflammatory cascade cycle in the wound site and promoted vascular and tissue regeneration. We posit that the COF-modified microalgae gel represents a promising strategy for the active in vivo delivery of therapeutics to the wound body in intensive care unit settings.

Microalgal biochar assisted simultaneous removal of particulate matter, formaldehyde, and total volatile organic compounds (TVOC's) from indoor air.

Biochar-based materials for air treatment have gained significant attention for removing health-detrimental volatile organic compounds (VOCs) and particulate matter (PM) in indoor air settings. However, high turnaround time, multiple pretreatment processes involved, and high pore size and low surface area (>10 µm, <100 m2 g-1) of lignocellulosic feedstocks demand alternative biochar feedstock material. Considering this, we designed a simple first-of-its-kind indoor air scrubbing material using diatoms-enriched microalgae biochar. In the present study, the microalgae were cultivated on waste anaerobic digestate (biogas slurry) and were pyrolyzed at three different temperatures: 300 °C (BC300), 500 °C (BC500), and 700 °C (BC700). The BC500 and BC700 showed the highest removal efficiencies (99 %) for total volatile organic carbons (TVOCs) and formaldehyde (HCHO) at concentrations of 1.22 mg m-3 HCHO and 8.57 mg m-3 TVOC compared to 50% efficiency obtained with commercially available surgical, cloth, and N95 masks. The biochar obtained showed a high Brunauer-Emmett-Teller (BET) surface area of 238 m2 g-1 (BC500) and 480 m2 g-1 (BC700) and an average pore size of 9-11 nm due to the mesoporous characteristic of diatom frustules. The comparatively poor performance of BC300 was due to lower surface area (150 m2 g-1) arising from incomplete organic removal, as evidenced by FESEM-EDX and FTIR. The high removal efficiencies in BC500 and BC700 were also attributed to the presence of reactive functional groups such as -OH and R-NH2. Concurrently, the average particulate matter (PM10, PM2.5, and PM1) removal efficiency for BC500 and BC 700 ranged between 66 and 82.69 %. The PM removal performance of BC500 and BC700 was lower (15-20%) than commercially available masks. Overall, the present study highlights the importance of diatoms (reactive Si) present inside the pores of microalgal biochar for enhanced removal of PM, TVOCs, and HCHO at temperatures above 500 °C. This complete approach signifies a step towards establishing a self-sustainable and circular process characterized by minimal waste generation for indoor air treatment.

A Holistic Approach to Circular Bioeconomy Through the Sustainable Utilization of Microalgal Biomass for Biofuel and Other Value-Added Products.

Emissions from transportation and industry primarily cause global warming, leading to floods, glacier melt, and rising seas. Widespread greenhouse gas emissions and resulting global warming pose significant risks to the environment, economy, and society. The need for alternative fuels drives the development of third-generation feedstocks: microalgae, seaweed, and cyanobacteria. These microalgae offer traits like rapid growth, high lipid content, non-competition with human food, and growth on non-arable land using brackish or waste water, making them promising for biofuel. These unique phototrophic organisms use sunlight, water, and carbon dioxide (CO2) to produce biofuels, biochemicals, and more. This review delves into the realm of microalgal biofuels, exploring contemporary methodologies employed for lipid extraction, significant value-added products, and the challenges inherent in their commercial-scale production. While the cost of microalgae bioproducts remains high, utilizing wastewater nutrients for cultivation could substantially cut production costs. Furthermore, this review summarizes the significance of biocircular economy approaches, which encompass the utilization of microalgal biomass as a feed supplement and biofertilizer, and biosorption of heavy metals and dyes. Besides, the discussion extends to the in-depth analysis and future prospects on the commercial potential of biofuel within the context of sustainable development. An economically efficient microalgae biorefinery should prioritize affordable nutrient inputs, efficient harvesting techniques, and the generation of valuable by-products.

Biogas Purification through the use of a Microalgae-Bacterial System in Semi-Industrial High Rate Algal Ponds.

In recent years, a number of technologies have emerged to purify biogas into biomethane. This purification entails a reduction in the concentration of polluting gases such as carbon dioxide and hydrogen sulfide to increase the content of methane. In this study, we used a microalgal cultivation technology to treat and purify biogas produced from organic waste from the swine industry to obtain ready-to-use biomethane. For cultivation and purification, two 22.2 m3 open-pond photobioreactors coupled with an absorption-desorption column system were set up in San Juan de los Lagos, Mexico. Several recirculation liquid/biogas ratios (L/G) were tested to obtain the highest removal efficiencies; other parameters, such as pH, dissolved oxygen (DO), temperature, and biomass growth, were measured. The most efficient L/Gs were 1.6 and 2.5, resulting in a treated biogas effluent with a composition of 6.8%vol and 6.6%vol in CO2, respectively, and removal efficiencies for H2S up to 98.9%, as well as maintaining O2 contamination values of less than 2%vol. We found that pH greatly determines CO2 removal, more so than L/G, during cultivation because of its participation in the photosynthetic process of microalgae and its ability to vary pH when solubilized due to its acidic nature. DO, and temperature oscillated as expected from the light-dark natural cycles of photosynthesis and the time of day, respectively. Biomass growth varied with CO2 and nutrient feeding as well as reactor harvesting; however, the trend remained primed for growth.

Photoheterotroph improved the growth and nutrient levels of Chlorella vulgaris and the related molecular mechanism.

Microalgae are rich in fatty acids, proteins, and other nutrients, which have gained the general attention of researchers all over the world. For the development of Chlorella vulgaris in food and feed industry, this study was conducted to investigate the differences in C. vulgaris' growth and nutritional components under different culture conditions (autotrophic, heterotrophic, photoheterotrophic) and the internal factors through cell counting in combination with transcriptome and nutrient analyses. The results showed that, under the photoheterotrophic condition, Chlorella's growth and the contents of lipid and protein were significantly higher than that under the heterotrophic condition, and the moisture content was lower than that under the heterotrophic condition. The saturated fatty acid content under the photoheterotrophic condition was the lowest, while the polyunsaturated fatty acid content was significantly higher than those under the other two conditions. There were 46,583 differentially expressed genes (DEGs), including 33,039 up-regulated DEGs (70.93%) and 13,544 down-regulated DEGs (29.07%), under the photoheterotrophic condition in comparison with the autotrophic condition. The fold change between the two conditions of samples of up-regulated genes was higher than that of the down-regulated genes. The KEGG enrichment showed that the up-regulated DEGs in the photoheterotrophic condition were significantly enriched in 5 pathways, including protein processing in endoplasmic reticulum pathway, photosynthesis pathway, photosynthesis-antenna protein pathway, endocytosis pathway, and phosphonate and phosphinate metabolism pathway. DEGs related to fatty acid metabolic pathways were significantly enriched in the fatty acid biosynthesis pathway and the biosynthesis of unsaturated fatty acid pathway. The qPCR analysis showed that the expression pattern of the selected genes was consistent with that of transcriptome analysis. The results of this study lay a theoretical foundation for the large-scale production of Chlorella and its application in food, feed, and biodiesel. KEY POINTS: • Nutrient levels under photoheterotrophic condition were higher than other conditions. • Six important pathways were discovered that affect changes in nutritional composition. • Explored genes encode important enzymes in the differential metabolic pathways.

In Situ Formed Microalgae-Integrated Living Hydrogel for Enhanced Tumor Starvation Therapy and Immunotherapy through Photosynthetic Oxygenation.

The effectiveness of various cancer therapies for solid tumors is substantially limited by the highly hypoxic tumor microenvironment (TME). Here, a microalgae-integrated living hydrogel (ACG gel) is developed to concurrently enhance hypoxia-constrained tumor starvation therapy and immunotherapy. The ACG gel is formed in situ following intratumoral injection of a biohybrid fluid composed of alginate, Chlorella sorokiniana, and glucose oxidase, facilitated by the crossing-linking between divalent ions within tumors and alginate. The microalgae Chlorella sorokiniana embedded in ACG gel generate abundant oxygen through photosynthesis, enhancing glucose oxidase-catalyzed glucose consumption and shifting the TME from immunosuppressive to immunopermissive status, thus reducing the tumor cell energy supply and boosting antitumor immunity. In murine 4T1 tumor models, the ACG gel significantly suppresses tumor growth and effectively prevents postoperative tumor recurrence. This study, leveraging microalgae as natural oxygenerators, provides a versatile and universal strategy for the development of oxygen-dependent tumor therapies.

Emulgel based on fish skin collagen-microalgae-silver increased neovascularization and re-epithelialization of full thickness burn in rats.

Deep skin burn represents a global morbidity and mortality problem, and the limitation of topical treatment agents has motivated research to development new formulations capable of preventing infections and accelerating healing. The aim of this work was to develop and characterize an emulgel based on collagen (COL) and gelatin (GEL) extracted from fish skin associated with Chlorella vulgaris extract (CE) and silver nitrate (AgNO3). COL and GEL were characterized by physicochemical and thermal analyses; and CE by electrophoresis and its antioxidant capacity. Three emulgels formulations were developed: COL (0.5%) + GEL (2.5%) (E1), COL+GEL+CE (1%) (E2), and COL+GEL+CE + AgNO3 (0.1%) (E3). All formulations were characterized by physicochemical, rheology assays, and preclinical analyses: cytotoxicity (in vitro) and healing potential using a burn model in rats. COL and GEL showed typical physicochemical characteristics, and CE presented 1.3 mg/mL of proteins and antioxidant activity of 76%. Emulgels presented a coherent physicochemical profile and pseudoplastic behavior. Preclinical analysis showed concentration-dependent cytotoxicity against fibroblast and keratinocytes. In addition, all emulgels induced similar percentages of wound contraction and complete wound closure in 28 days. The histopathological analysis showed higher scores for polymorphonuclear cells to E1 and greater neovascularization and re-epithelialization to E3. Then, E3 formulation has potential to improve burn healing, although its use in a clinical setting requires further studies.

A comparison of two gene regions for assessing community composition of eukaryotic marine microalgae from coastal ecosystems.

Two gene regions commonly used to characterise the diversity of eukaryotic communities using metabarcoding are the 18S ribosomal DNA V4 and V9 gene regions. We assessed the effectiveness of these two regions for characterising diverisity of coastal eukaryotic microalgae communities (EMCs) from tropical and temperate sites. We binned amplicon sequence variants (ASVs) into the high level taxonomic groups: dinoflagellates, pennate diatoms, radial centric diatoms, polar centric diatoms, chlorophytes, haptophytes and 'other microalgae'. When V4 and V9 generated ASV abundances were compared, the V9 region generated a higher number of raw reads, captured more diversity from all high level taxonomic groups and was more closely aligned with the community composition determined using light microscopy. The V4 region did resolve more ASVs to a deeper taxonomic resolution within the dinoflagellates, but did not effectively resolve other major taxonomic divisions. When characterising these communities via metabarcoding, the use of multiple gene regions is recommended, but the V9 gene region can be used in isolation to provide high-level community biodiversity to reflect relative abundances within groups. This approach reduces the cost of sequencing multiple gene regions whilst still providing important baseline ecosystem function information.