Search

Global release of plastics exerts various impacts on the ecological cycle, particularly on primary photosynthesis, while the impacts of plastic additives are unknown. As a carrier of fluorescent brightener, plastic particles co-modify Chlorella pyrenoidosa (C. pyrenoidosa) growth and its photosynthetic parameters. In general, adding to the oxidative damage induced by polystyrene, fluorescent brightener-doped polystyrene produces stronger visible light and the amount of negative charge is more likely to cause photodamage in C. pyrenoidosa leading to higher energy dissipation through conditioning than in the control group with a date of ETR (II) inhibition rate of 33 %, Fv/Fm inhibition rate of 8.3 % and Pm inhibition rate of 48.8 %. To elucidate the ecological effect of fluorescent brightener doping in plastic particles, a machine learning method is performed to establish a Gradient Boosting Machine model for predicting the impact of environmental factors on algal growth. Upon validation, the model achieved an average fitting degree of 88 %. Relative concentration of plastic particles and algae claimed the most significant factor by interpretability analysis of the machine learning. Additionally, both Gradient Boosting Machine prediction and experimental results indicate a matching result that plastic additives have an inhibitive effect on algal growth., 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.)

Oxidative stress is a universal interpretation for the toxicity mechanism of nanoplastics to microalgae. However, there is a lack of deeper insight into the regulation mechanism in microalgae response to oxidative stress, thus affecting the prevention and control for nanoplastics hazard. The integrated analysis of transcriptomics and metabolomics was employed to investigate the mechanism for the oxidative stress response of Chlorella pyrenoidosa to nanoplastics and subsequently lock the according core pathways and driver genes induced. Results indicated that the linoleic acid metabolism, glycine (Gly)-serine (Ser)-threonine (Thr) metabolism, and arginine and proline metabolism pathways of C. pyrenoidosa were collectively involved in oxidative stress. The analysis of linoleic acid metabolism suggested that nanoplastics prompted algal cells to secrete more allelochemicals, thereby leading to destroy the immune system of cells. Gly-Ser-Thr metabolism and arginine and proline metabolism pathways were core pathways involved in algal regulation of cell membrane function and antioxidant system. Key genes, such as LOX2.3, SHM1, TRPA1, and proC1, are drivers of regulating the oxidative stress of algae cells. This investigation lays the foundation for future applications of gene editing technology to limit the hazards of nanoplastics on aquatic organism., Competing Interests: Declaration of Competing interest This paper is our unpublished work and it has not been submitted to any other journal for reviews. Thus, it has not been previously published, in whole or in part, and it is not under consideration by any other journal. Additionally, I confirm that both authors are aware of, and accept responsibility for, the manuscript. There is no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microplastics (MPs) are emerging pollutants, causing potential threats to aquatic ecosystems and serious concern in aggregating with microalgae (critical primary producers). When entering water bodies, MPs are expected to sink below the water surface and disperse into varying water compartments with different light intensities. However, how light influences the aggregation processes of algal cells onto MPs and the associated molecular coupling mechanisms and derivative risks remain poorly understood. Herein, we investigated the aggregation behavior between polystyrene microplastics (mPS, 10 µm) and Chlorella pyrenoidosa under low (LL, 15 μmol·m -2 ·s -1 ), normal (NL, 55 μmol·m -2 ·s -1 ), and high light (HL, 150 μmol·m -2 ·s -1 ) conditions from integrated in vivo and in silico assays. The results indicated that under LL, the mPS particles primarily existed independently, whereas under NL and HL, C. pyrenoidosa tightly bounded to mPS by secreting more protein-rich extracellular polymeric substances. Infrared spectroscopy analysis and density functional theory calculation revealed that the aggregation formation was driven by non-covalent interaction involving van der Waals force and hydrogen bond. These processes subsequently enhanced the deposition and adherence capacity of mPS and relieved its phytotoxicity. Overall, our findings advance the practical and theoretical understanding of the ecological impacts of MPs in complex aquatic environments., 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.)

Microalgal biomass is gaining increasing attention to produce high-value co-products. This study proposes integrating Chlorella microalgal biomass into a zero-waste biorefining system, aiming to produce biodiesel and biofertilizer. It investigates optimal conditions for ultrasound-assisted deep eutectic solvent (DES) pretreatment and lipid recovery to enhance the extraction of lipids. Optimal DES pretreatment was identified as a 1.6:1 acetic acid-to-choline chloride molar ratio, 0.36 g biomass loading, and 2.50 min of pretreatment. Lipid recovery succeeded with a 10-minute extraction time and a 1:3 methanol-to-butanol volume ratio. These conditions yielded biodiesel-quality lipids at 139.52 mg/g microalgal biomass with superior fuel characteristics. The de-oiled microalgal biomass residue exhibited promise as a lettuce biofertilizer, enhancing photosynthetic pigments but potentially reducing yields by 40 %. The study also notes changes in rhizosphere microbial communities, indicating both stimulatory and inhibitory effects on beneficial microbes. This study has the potential to enhance sustainability in energy, agriculture, and the environment., 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.)

This study investigated the bioenergy potential of Chlorella pyrenoidosa (CP) for use as fuel and carbon material through chemical and thermal characterization. The thermo-kinetic characteristics of Chlorella pyrenoidosa were assessed using isoconversional, linear regression, and non-linear regression approaches. The physicochemical analysis revealed high carbon (53.1 %), volatile (69.35 %), and low moisture (2.19 %), ash content (3.42 %). The results indicated that the non-linear model fitting method was the most accurate with the approximated activation energy (E α ) and pre-exponential Arrhenius constant (Ln A) were 124.92 ± 2.74 kJ/mol and 23.38 ± 4.63 min -1 , respectively. Additionally, the inclusion of sodium bicarbonate resulted in a significant increase in BET surface area. FTIR analysis revealed several functional groups beneficial for carbon material, while XRD analysis showed a broad peak correlated with an amorphous structure. This study highlighted the potential of Chlorella pyrenoidosa biomass for various applications, including carbon material and renewable fuel., 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.)

In this study, Chlorella and Scenedesmus were inoculated in biogas slurry medium with initial inoculum (OD 680 ) of 0.05, 0.1, 0.2, and 0.3, respectively, and 5% CO 2 was continuously injected. The study aimed to examine the carbon sequestration capacity of Chlorella and Scenedesmus , as well as the effectiveness of removing pollutants such as TN, TP, and COD in biogas slurry medium. Additionally, an economic efficiency analysis of energy consumption was conducted. The group with an initial inoculum (OD 680 ) of 0.3 for both types of microalgae exhibited better tolerance to pollutants, entered the logarithmic growth stage earlier, promoted nutrient removal, achieved higher energy efficiency, and reduced carbon emissions compared to the other groups. The highest carbon sequestration rates were 18.03% for Chlorella and 11.05% for Scenedesmus . Furthermore, Chlorella demonstrated corresponding nutrient removal efficiencies of 83.03% for TN, 99.84% for TP, and 90.06% for COD, while Scenedesmus exhibited removal efficiencies of 66.35% for TN, 98.74% for TP, and 77.71% for COD. The highest energy efficiency for pollutants and CO 2 removal rates for Chlorella were 49.51 ± 2.20 and 9.91 ± 0.44 USD -1 , respectively. In conclusion, the findings demonstrate the feasibility of using microalgae for simultaneous purification of biogas and biogas slurry.

This study aimed to assess the antioxidant activity of golden chlorella (GoC) and grape pomace (GrP) extracts both in vitro and in pea protein-based extrudates. We hypothesized that GoC/GrP would limit oxidation of proteins in the extrudates compared with commercial antioxidants. The results showed that GoC extract was effective in metal chelation and GrP extract possessed excellent radical scavenging activity and reducing power. Protein oxidation inevitably occurred after low-moisture extrusion in terms of elevated level of protein carbonyls and the gradual loss of thiols. LC-MS/MS revealed that the monoxidation and 4-hydroxynonenal adduction were the major oxidative modifications, and legumin was the most susceptible globulin for oxidation. The GoC/GrP extracts effectively retarded the oxidation progress in extrudates by lower intensity of oxidized peptides, whereas protein electrophoretic profiles remained unaffected. This study highlighted the great potential of GoC/GrP as natural antioxidants in plant-based foods., 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.)

Sulfadiazine (SDZ) is a kind of anti-degradable antibiotics that is commonly found in wastewater, but its removal mechanism and transformation pathway remain unclear in microalgal systems. This study investigated the effects of initial algae concentration and SDZ-induced stress on microalgal growth metabolism, SDZ removal efficiency, and transformation pathways during Chlorella sp. cultivation. Results showed that SDZ had an inhibitory effect on the growth of microalgae, and increasing the initial algal biomass could alleviate the inhibitory effect of SDZ. When the initial algal biomass of Chlorella sp. was increased to 0.25 g L -1 , the SDZ removal rate could reach 53.27%-89.07%. The higher the initial algal biomass, the higher the SOD activity of microalgae, and the better the protective effect on microalgae, which was one of the reasons for the increase in SDZ removal efficiency. Meanwhile, SDZ stress causes changes in photosynthetic pigments, lipids, total sugars and protein content of Chlorella sp. in response to environmental changes. The main degradation mechanisms of SDZ by Chlorella sp. were biodegradation (37.82%) and photodegradation (23%). Most of the degradation products of SDZ were less toxic than the parent compound, and the green algae were highly susceptible to SDZ and its degradation products. The findings from this study offered valuable insights into the tradeoffs between accumulating microalgal biomass and antibiotic toxic risks during wastewater treatment, providing essential direction for the advancement in future research and full-scale application., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:SHAN LIU reports financial support was provided by Shandong Provincial Natural Science Foundation. SHAN LIU reports financial support was provided by Yantai Educational-Local Synthetic Development Project. SHAN LIU reports financial support was provided by Key Laboratory for Crop and Animal Integrated Farming, Ministry of Agriculture and Rural Affairs. If there are other authors, they 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.)

The degradation of isopyrazam in soils was investigated through kinetics, microbial contributions, and transformation products (TPs). Then the acute toxicity of isopyrazam and its TP to Chlorella pyrenoidosa was explored. The half-lives of isopyrazam in cinnamon soil, red soil, and black soil were 82.2, 141.7, and 120.3 days, respectively. A strain ( Bacillus sp. A01) isolated from cinnamon soil could degrade 72.9% of isopyrazam at 10 mg/L after 6 days in a Luria-Bertani medium. Six TPs were observed with Bacillus sp. A01, and three of them were found in soil as well. Through the inhibition of cytochrome P450 enzymes, the production of oxidized isopyrazam was blocked. Microbial mediated hydroxylation, epoxidation, and dehydration were the main degradation pathways of isopyrazam. The acute toxicity results showed that the EC 50 of 3-(difluoromethyl)- N -(9-(2-hydroxypropan-2-yl)-1,2,3,4-tetrahydro-1,4-methanonaphthalen-6-yl)-1-methyl-1 H -pyrazole-4-carboxamide to Chlorella pyrenoidosa was 40 times higher than that of the parent. This work provides new insights for understanding the degradation behavior of isopyrazam in soil.

Various bisphenols (BPs) have been frequently detected in the aquatic environment and coexist in the form of mixtures with potential huge risks. As we all know, food chain is a media by which BPs mixtures and their mixtures probably enter the organisms at different trophic levels due to their environmental persistence. As a result, the concentrations of BPs and their mixtures may continuously magnify to varying degrees, which can produce higher risks to different levels of organisms, and even human health. However, the related researches about mixtures are few due to the complexity of mixtures. So, the ternary BP mixtures were designed by the uniform design ray method using bisphenol A (BPA), bisphenol S (BPS) and bisphenol F (BPF) to investigate their food chain effects including bioconcentration and biomagnification. Here, Chlorella pyrenoidosa (C. pyrenoidosa) and Daphnia magna (D. magna) were selected to construct a food chain. The toxic effects of single BPs and their mixtures were also systematically investigated by the time-dependent microplate toxicity analysis (t-MTA) method. Toxicity interaction within the ternary mixture was analyzed by the concentration addition model (CA) and the deviation from the CA model (dCA). The results show that the C. pyrenoidosa and D. magna had obvious bioconcentration and biomagnification effects on BPs and their mixture. The mixture had the potential to enrich at higher nutrient levels. And BPF had the largest bioconcentration effect (BCF 1  = 481.86, BCF 2  = 772.02) and biomagnification effect (BMF = 1.6). Three BPs were toxic to C. pyrenoidosa by destroying algal cells and decreasing protein and chlorophyll contents, and their toxicity order was BPF > BPA > BPS. Moreover, their ternary mixture exhibits synergism with time/concentration-dependency. The obtained results are of significant reference value for objectively and accurately assessing the ecological and environmental risks of bisphenol pollutants., 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.)

Background: When subject to stress conditions such as nutrient limitation microalgae accumulate triacylglycerol (TAG). Fatty acid, a substrate for TAG synthesis is derived from de novo synthesis or by membrane remodeling. The model industrial alga Chlorellasorokiniana accumulates TAG and other storage compounds under nitrogen (N)-limited growth. Molecular mechanisms underlying these processes are still to be elucidated., Result: Previously we used transcriptomics to explore the regulation of TAG synthesis in C. sorokiniana. Surprisingly, our analysis showed that the expression of several key genes encoding enzymes involved in plastidic fatty acid synthesis are significantly repressed. Metabolic labeling with radiolabeled acetate showed that de novo fatty acid synthesis is indeed downregulated under N-limitation. Likewise, inhibition of the Target of Rapamycin kinase (TOR), a key regulator of metabolism and growth, decreased fatty acid synthesis. We compared the changes in proteins and phosphoprotein abundance using a proteomics and phosphoproteomics approach in C. sorokiniana cells under N-limitation or TOR inhibition and found extensive overlap between the N-limited and TOR-inhibited conditions. We also identified changes in the phosphorylation status of TOR complex proteins, TOR-kinase, and RAPTOR, under N-limitation. This indicates that TOR signaling is altered in a nitrogen-dependent manner. We find that TOR-mediated metabolic remodeling of fatty acid synthesis under N-limitation is conserved in the chlorophyte algae Chlorella sorokiniana and Chlamydomonas reinhardtii., Conclusion: Our results indicate that under N-limitation there is significant metabolic remodeling, including fatty acid synthesis, mediated by TOR signaling. This process is conserved across chlorophyte algae. Using proteomic and phosphoproteomic analysis, we show that N-limitation affects TOR signaling and this in-turn affects the metabolic status of the cells. This study presents a link between N-limitation, TOR signaling and fatty acid synthesis in green-lineage., (© 2024. The Author(s).)

Ubiquitous distribution of pharmaceutical contaminants in environment has caused unexpected adverse effects on ecological organisms; however, how microorganisms recover from their toxicities remains largely unknown. In this study, we comprehensively investigated the effect of a representative pollutant, doxylamine (DOX) on a freshwater microalgal species, Chlorella sp. by analyzing the growth patterns, biochemical changes (total chlorophyll, carotenoid, carbohydrate, protein, and antioxidant enzymes), and transcriptomics. We found toxicity of DOX on Chlorella sp. was mainly caused by disrupting synthesis of ribosomes in nucleolus, and r/t RNA binding and processing. Intriguingly, additional bicarbonate enhanced the toxicity of DOX with decreasing the half-maximum effective concentrations from 15.34 mg L -1 to 4.63 mg L -1 , which can be caused by inhibiting fatty acid oxidation and amino acid metabolism. Microalgal cells can recover from this stress via upregulating antioxidant enzymatic activities to neutralize oxidative stresses, and photosynthetic pathways and nitrogen metabolism to supply more energies and cellular signaling molecules. This study extended our understanding on how microalgae can recover from chemical toxicity, and also emphasized the effect of environmental factors on the toxicity of these contaminants on aquatic microorganisms., 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 microalgae Chlorella sorokiniana was used for the treatment of winery wastewater (WWW). Batch experiments were initially conducted to investigate how biomass acclimatization in different media, dilution of wastewater, and addition of ammonium nitrogen (NH 4 -N) affect the growth of microalgae and the removal of major pollutants. Afterwards, two sequencing batch reactor (SBR) systems were tested applying different configurations and hydraulic retention times. The biomass collected at the end of the experiments was characterized for proteins, lipids, carbohydrates, amino acid profile, and the existence of lutein, β-carotene, chlorophyll a, and tocopherols. Batch experiments showed that Chlorella sorokiniana acclimatization to urban wastewater enhanced the removal of NH 4 -N and total phosphorus (TP). The operation of a two-stage SBR system achieved COD and NH 4 -N removal equal to 85 ± 9% and 91 ± 20%, respectively, while the use of a single-stage system feeding with anaerobically pretreated WWW resulted to COD and NH 4 -N removal of 78 ± 9% and 95 ± 9%, respectively. Analyses of biomass showed higher protein content (up to 58.8%) in batch experiments with NH 4 -N addition as well as in SBR experiments. The cultivation of microalgae under SBR conditions enhanced the production of pigments and tocopherols. The maximum concentrations of 1075 mg kg -1 , 45.5 mg kg -1 , and 131.2 mg kg -1 were achieved for lutein, β-carotene, and tocopherols, respectively, in the one-stage system. Our findings suggested that Chlorella sorokiniana cultivation in WWW not only removed nutrients from WWW but also could potentially serve for the production of value-added ingredients used in food industry, cosmetics, and animal feedstock., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Microalgae are known to be the richest natural source of polysaccharides. The study aimed to evaluate the ability of microalgae from the Chlorella sp. genus to synthesize polysaccharides. Brody & Emerson max medium proved to be the most effective; the average cell content in the culture fluid at the beginning and at the end of cultivation for IPPAS Chlorella pyrenoidosa Chick was 1.23 ± 0.03 g/L and 1.71 ± 0.20 g/L, respectively. With a high average dry weight of IPPAS Chlorella pyrenoidosa Chick (4.45 ± 0.10 g/L), it produced the least amount of neutral sugars (0.75 ± 0.02 g/L) and uronic acids (0.14 ± 0.01 mg/L). The microalga IPPAS Chlorella vulgaris with the lowest average dry weight (1.18 ± 0.03 g/L) produced 0.80 ± 0.02 g/L of neutral sugars and 0.17 ± 0.01 mg/L of uronic acids. Microalgal polysaccharides have the potential to be used as a source for biologically active food additives, as they contain various types of polysaccharides that can be beneficial to human health., 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-based biotechnology holds significant potential for addressing dual challenges of phosphorus removal and recovery from wastewater; however, the removal mechanism and metabolic adaptation of microalgae to dissolved organic phosphorus (DOP) are still unclear. This study investigated the removal mechanisms and metabolomic responses of the Chlorella pyrenoidosa to different DOP forms, including adenosine triphosphate (ATP), glucose-6-phosphate (G-6-P), and β-glycerophosphate (β-GP). The results showed C. pyrenoidosa could efficiently take up above 96% DOP through direct transport and post-hydrolysis pathways. The uptake of inorganic phosphorus (IP) followed pseudo first order kinetic model, while DOP followed pseudo second order kinetic model. Metabolite profiling revealed substantial alterations in central carbon metabolism depending on the DOP source. G-6-P upregulated glycolytic and TCA cycle intermediates, reflecting enhanced carbohydrates, amino acids and nucleotides biosynthesis. In contrast, ATP down-regulated carbohydrate and purine metabolism, inhibiting sustainable growth of microalgae. This study offers theoretical support for phosphorus-containing wastewater treatment using microalgae., 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.)

Energy is a crucial entity for the development and it has various alternative forms of energy sources. Recently, the synthesis of nanoparticles using benign biocatalyst has attracted increased attention. In this study, silver nanoparticles were synthesized and characterized using Azadirachta indica plant-derived phytochemical as the reducing agent. Biomass of the microalga Chlorella sp. cultivated in BG11 medium increased after exposure to low concentrations of up to 0.48 mg L -1 AgNPs. In addition, algal cells treated with 0.24 mg L -1 AgNPs and cultivated in BG11 0 medium which contained no nitrogen source showed the highest hydrogen yield of 10.8 mmol L -1 , whereas the untreated cells under the same conditions showed very low hydrogen yield of 0.003 mmol L -1 . The enhanced hydrogen production observed in the treated cells was consistent with an increase in hydrogenase activity. Treatment of BG11 0 grown cells with low concentration of green synthesized AgNPs at 0.24 mg L -1 enhanced hydrogenase activity with a 5-fold increase of enzyme activity compared to untreated BG11 0 grown cells. In addition, to improve photolytic water splitting efficiency for hydrogen production, cells treated with AgNPs at 0.24 mg L -1 showed highest oxygen evolution signifying improvement in photosynthesis. The silver nanoparticles synthesized using phytochemicals derived from plant enhanced both microalgal biomass and hydrogen production with an added advantage of CO 2 reduction which could be achieved due to an increase in biomass. Hence, treating microalgae with nanoparticles provided a promising strategy to reduce the atmospheric carbon dioxide as well as increasing production of hydrogen as clean energy., 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.)

This study analyzed the effects of Ca 2+ metal ions among culture medium components on the Chlorella sorokiniana strain DSCG150 strain cell growth. The C. sorokiniana strain DSCG150 grew based on a multiple fission cell cycle and growth became stagnant in the absence of metal ions in the medium, particularly Ca 2+ . Flow cytometry and confocal microscopic image analysis results showed that in the absence of Ca 2+ , cell growth became stagnant as the cells accumulated into four autospores and could not transform into daughter cells. Genetic analysis showed that the absence of Ca 2+ caused upregulation of calmodulin ( calA ) and cell division control protein 2 ( CDC2_1 ) genes, and downregulation of origin of replication complex subunit 6 ( ORC6 ) and dual specificity protein phosphatase CDC14A ( CDC14A ) genes. Analysis of gene expression patterns by qRT-PCR showed that the absence of Ca 2+ did not affect cell cycle progression up to 4n autospore, but it inhibited Chlorella cell fission (liberation of autospores). The addition of Ca 2+ to cells cultivated in the absence of Ca 2+ resulted in an increase in n cell population, leading to the resumption of C. sorokiniana growth. These findings suggest that Ca 2+ plays a crucial role in the fission process in Chlorella .

Chlorella sorokiniana , isolated from a pond adjacent to a cement plant, was cultured using flue gas collected directly from kiln emissions using 20 L and 25000 L photobioreactors. Lipids, proteins, and polysaccharides were analyzed to understand their overall composition for potential applications. The lipid content ranged from 17.97% to 21.54% of the dry biomass, with carotenoid concentrations between 8.4 and 9.2 mg/g. Lutein accounted for 55% of the total carotenoids. LC/MS analysis led to the identification of 71 intact triacylglycerols, 8 lysophosphatidylcholines, 10 phosphatidylcholines, 9 monogalactosyldiacylglycerols, 12 digalactosyldiacylglycerols, and 1 sulfoquinovosyl diacylglycerol. Palmitic acid, oleic acid, linoleic acid, and α-linolenic acid were the main fatty acids. Polyunsaturated fatty acid covers ≥ 56% of total fatty acids. Protein isolates and polysaccharides were also extracted. Protein purity was determined to be ≥75% by amino acid analysis, with all essential amino acids present. Monomer analysis of polysaccharides suggested that they are composed of mainly D-(+)-mannose, D-(+)-galactose, and D-(+)-glucose. The results demonstrate that there is no adverse effect on the metabolite profile of C. sorokiniana biomass cultured using flue gas as the primary carbon source, revealing the possibility of utilizing such algal biomass in industrial applications such as animal feed, sources of cosmeceuticals, and as biofuel.

The phycosphere is an essential ecological niche for the proliferation of antibiotic resistance genes (ARGs). However, how ARGs' potential hosts change and the driving mechanism of metabolites under antibiotic stress in the phycosphere have seldom been researched. We investigated the response of Chlorella pyrenoidosa and the structure and abundance of free-living (FL) and particle-attached (PA) bacteria, ARGs, and metabolites under sulfadiazine by using real-time quantitative PCR, 16 S rRNA high-throughput. The linkage of key bacterial communities, ARGs, and metabolites through correlations was established. Through analysis of physiological indicators, Chlorella pyrenoidosa displayed a pattern of "low-dose promotion and high-dose inhibition" under antibiotic stress. ARGs were enriched in the PA treatment groups by 117 %. At the phylum level, Proteobacteria, Bacteroidetes, and Actinobacteria as potential hosts for ARGs. At the genus level, potential hosts included Sphingopyxis, SM1A02, Aquimonas, Vitellibacter, and Proteiniphilum. Middle and high antibiotic concentrations induced the secretion of metabolites closely related to potential hosts by algae, such as phytosphingosine, Lysophosphatidylcholine, and α-Linolenic acid. Therefore, changes in bacterial communities indirectly influenced the distribution of ARGs through alterations in metabolic products. These findings offer essential details about the mechanisms behind the spread and proliferation of ARGs in the phycosphere., 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.)

Protists, including ciliates retain crystals in their cytoplasm. However, their functions and properties remain unclear. To comparatively analyze the crystals of Paramecium bursaria, a ciliate, associated with and without the endosymbiotic Chlorella variabilis, we investigated the isolated crystals using a light microscope and analyzed their length and solubility. A negligible number of crystals was found in P. bursaria cells harboring symbiotic algae. The average crystal length in alga-free and algae-reduced cells was about 6.8 μm and 14.4 μm, respectively. The crystals of alga-free cells were spherical, whereas those of algae-reduced cells were angular in shape. The crystals of alga-free cells immediately dissolved in acids and bases, but not in water or organic solvents, and were stable at - 20 °C for more than 3 weeks. This study, for the first time, reveals that the characteristics of crystals present in the cytoplasm of P. bursaria vary greatly depending on the amount of symbiotic algae., (© 2024. The Author(s).)

Algal biomass is a viable source of chemicals and metabolites for various energy, nutritional, medicinal and agricultural uses. While stresses have commonly been used to induce metabolite accumulation in microalgae in attempts to enhance high-value product yields, this is often very detrimental to growth. Therefore, understanding how to modify metabolism without deleterious consequences is highly beneficial. We demonstrate that low-doses (1-5 Gy) of ionizing radiation in the X-ray range induces a non-toxic, hormetic response in microalgae to promote metabolic activation. We identify specific radiation exposure parameters that give reproducible metabolic responses in Chlorella sorokiniana caused by transcriptional changes. This includes up-regulation of >30 lipid metabolism genes, such as genes encoding an acetyl-CoA carboxylase subunit, phosphatidic acid phosphatase, lysophosphatidic acid acyltransferase, and diacylglycerol acyltransferase. The outcome is an increased lipid yield in stationary phase cultures by 25% in just 24 hours, without any negative effects on cell viability or biomass., (© 2024. The Author(s).)

Microalgae can promote antibiotic removal, which has attracted growing attention. However, its synergistic removal performance with bacteria in antibiotic pollutants is still poorly understood. In this study, firstly, we selected two green algae (Dictyosphaerium sp. and Chlorella sp.) and exposed them to Enrofloxacin (ENR) to observe their extracellular polysaccharides (EPS) concentration dynamic and the removal of antibiotics. Secondly, EPS was extracted and added to in situ lake water (no algae) to investigate its combined effect with bacteria. The results indicate that both Dictyosphaerium sp. and Chlorella sp. exhibited high tolerance to ENR stress. When the biomass of microalgae was low, ENR could significantly stimulate algae to produce EPS. The removal rates of Dictyosphaerium sp. and Chlorella sp. were 15.8% and 10.5%, respectively. The addition of EPS can both alter the microbial community structure in the lake water and promote the removal of ENR. The LEfSe analysis showed that there were significant differences in the microbial marker taxa, which promoted the increase of special functional bacteria for decomposing ENR, between the EPS-added group and the control group. The EPS of Dictyosphaerium sp. increased the abundance of Moraxellaceae and Spirosomaceae, while the EPS of Chlorella sp. increased the abundance of Sphingomonadaceae and Microbacteriaceae. Under the synergistic effect, Chlorella sp. achieved a maximum removal rate of 24.2%, while Dictyosphaerium sp. achieved a maximum removal rate of 28.9%. Our study provides new insights into the removal performance and mechanism of antibiotics by freshwater microalgae in water bodies and contribute to the development of more effective water treatment strategies., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Main Conclusion: Nitrogen stress altered important lipid parameters and related genes in Chlorella pyrenoidosa via ROS and Ca 2+ signaling. The mutual interference between ROS and Ca 2+ signaling was also uncovered. The changed mechanisms of lipid parameters (especially lipid classes and unsaturation of fatty acids) in microalgae are not completely well known under nitrogen stress. Therefore, Chlorella pyrenoidosa was exposed to 0, 0.5, 1 and 1.5 g L -1 NaNO 3 for 4 days. Then, the physiological and biochemical changes were measured. It was shown that the total lipid contents, neutral lipid ratios as well as their related genes (accD and DGAT) increased obviously while the polar lipid ratios, degrees of unsaturation as well as their related genes (PGP and desC) decreased significantly in nitrogen stress groups. The obvious correlations supported that gene expressions should be the necessary pathways to regulate the lipid changes in C. pyrenoidosa under nitrogen stress. The changes in ROS and Ca 2+ signaling as well as their significant correlations with corresponding genes and lipid parameters were analyzed. The results suggested that ROS and Ca 2+ may regulate these gene expressions and lipid changes in C. pyrenoidosa under nitrogen stress conditions. This was verified by the subordinate tests with an ROS inhibitor and calcium reagents. It also uncovered the clues of mutual interference between ROS and Ca 2+ signaling. To summarize, this study revealed the signaling pathways of important lipid changes in microalgae under N stress., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Antibiotics are ubiquitously present in aquatic environments, posing a serious ecological risk to aquatic ecosystems. However, the effects of antibiotics on the photosynthetic light reactions of freshwater algae and the underlying mechanisms are relatively less understood. In this study, the effects of 4 representative antibiotics (clarithromycin, enrofloxacin, tetracycline, and sulfamethazine) on a freshwater alga (Chlorella pyrenoidosa) and the associated mechanisms, primarily focusing on key regulators of the photosynthetic light reactions, were evaluated. Algae were exposed to different concentrations of clarithromycin (0.0-0.3 mg/L), enrofloxacin (0.0-30.0 mg/L), tetracycline (0.0-10.0 mg/L), and sulfamethazine (0.0-50.0 mg/L) for 7 days. The results showed that the 4 antibiotics inhibited the growth, the photosynthetic pigment contents, and the activity of antioxidant enzymes. In addition, exposure to clarithromycin caused a 118.4 % increase in malondialdehyde (MDA) levels at 0.3 mg/L. Furthermore, the transcripts of genes for the adenosine triphosphate (ATP) - dependent chloroplast proteases (ftsH and clpP), genes in photosystem II (psbA, psbB, and psbC), genes related to ATP synthase (atpA, atpB, and atpH), and petA (related to cytochrome b6/f complex) were altered by clarithromycin. This study contributes to a better understanding of the risk of antibiotics on primary producers in aquatic environment., 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.)

Prior research has emphasized the potential of microalgae in biodiesel production, driven by their ability to replace fossil fuels. However, the significant costs associated with microalgae cultivation present a major obstacle to scaling up production. This study aims to develop an eco-friendly microalgae cultivation system by integrating carbon dioxide from flue gas emissions with an affordable photobioreactor, providing a sustainable biomass production. The research evaluates the growth performance of Chlorella sorokiniana and Chlorella vulgaris across this integrated system for biomass and lipid production. Results indicate substantial biomass yields of 1.97 and 1.84 g/L, with lipid contents of 35 % and 41 % for C. sorokiniana and C. vulgaris, respectively. The macrobubble photobioreactor demonstrates high potential for microalgae biomass and lipid production, yielding quality fatty acid methyl esters such as palmitic, linoleic and stearic. This study presents an environmentally friendly system for efficient microalgae cultivation, generating lipid-rich biomass suitable for biodiesel production., 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.)

Further treatment of secondary effluents before their discharge into the receiving water bodies could alleviate water eutrophication. In this study, the Chlorella proteinosa was cultured in a membrane photobioreactor to further remove nitrogen from the secondary effluents. The effect of hydraulic retention time (HRT) on microalgae biomass yields and nutrient removal was studied. The results showed that soluble algal products concentration reduced in the suspension at low HRT, thereby alleviating microalgal growth inhibition. In addition, the lower HRT reduced the nitrogen limitation for Chlorella proteinosa's growth through the phase-out of nitrogen-related functional bacteria. As a result, the productivity for Chlorella proteinosa increased from 6.12 mg/L/day at an HRT of 24 hr to 20.18 mg/L/day at an HRT of 8 hr. The highest removal rates of 19.7 mg/L/day, 23.8 mg/L/day, and 105.4 mg/L/day were achieved at an HRT of 8 hr for total nitrogen (TN), ammonia, and chemical oxygen demand (COD), respectively. However, in terms of removal rate, TN and COD were the largest when HRT is 24 hr, which were 74.5% and 82.6% respectively. The maximum removal rate of ammonia nitrogen was 99.2% when HRT was 8 hr., 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 © 2023. Published by Elsevier B.V.)

Tetracycline (TC) and ciprofloxacin (CF) induce a synergistic effect that alters the biochemical composition, leading to a decrease in the growth and photosynthetic efficiency of microalgae. But the current study provides a novel insight into stress-inducing techniques that trigger a change in macromolecules, leading to an increase in the bioenergy potential and pathogen resistance of Chlorella variabilis biofilm. The study revealed that in a closed system, a light intensity of 167 μmol/m 2 /s causes 93.5% degradation of TC and 16% degradation of CF after 7 days of exposure, hence availing the products for utilization by C. variabilis biofilm. The resistance to pathogens invasion was linked to 85% and 40% increase in the expression level of photosystem II oxygen-evolving enhancer protein 3 (PsbQ), and mitogen activated kinase (MAK) respectively. The results also indicate that a surge in light intensity triggers 49% increase in the expression level of lysophosphatidylcholine (LPC) (18:2), which is an important lipidomics that can easily undergo transesterification into bioenergy. The thermogravimetric result indicates that the biomass sample of C. variabilis biofilm cultivated under light intensity of 167 μmol/m2/s produces a higher residual mass of 45.5% and 57.5 under air and inert conditions, respectively. The Fourier transform infrared (FTIR) indicates a slight shift in the major functional groups, while the energy-dispersive X-ray spectroscopy (SEM-EDS) and X-ray fluorescence (XRF) indicate clear differences in the morphology and elemental composition of the biofilm biomass in support of the increase bioenergy potential of C. variabilis biofilm. The current study provides a vital understanding of a innovative method of cultivation of C. variabilis biofilm, which is resistant to pathogens and controls the balance between fatty acid and TAG synthesis leading to surge in bioenergy potential and environmental 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 Inc.)

The current study investigated the effect of iron oxide nanoparticles (IONPs) on mixotrophic microalgae cultivation in wastewater for biofuel production. Optimal IONPs doses of 10 and 20 mg L -1 increased Chlorella pyrenoidosa growth by 16% and lipid accumulation by 53 %, respectively, compared with the control group. Conversely, the protein content declined drastically, while carbohydrates remained relatively unchanged. A maximum of 15% rise in biomass growth was observed for Chlorella sorokiniana IITRF at an IONPs dose of 20 mg L -1 , with no significant variation in biochemical composition. Microalgae grown under mixotrophic conditions with IONPs in a biofilm reactor were more suitable for biogas production than biodiesel, increasing biogas and methane content by 38 and 48%, respectively. The findings suggest that low doses of IONPs can enhance microalgal biomass, biogas production and methane content. Further, metabolomics studies are warranted to investigate the interaction between microalgae and nanoparticles to achieve high-quality biodiesel., 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.)

Zinc oxide nanoparticles (ZnO NPs) expedite the conversion of organic phosphorus (OP) into PO 4 -P (Pi), facilitating phosphorus (P) absorption by algae. Our study explored the mechanisms of converting OP (2-aminoethylphosphonic acid (AEP) and β-glycerol phosphate (β-GP)) into Pi in Chlorella pyrenoidosa under P deficiency with sunscreen and ZnO NPs. Cell density followed the order of K 2 HPO 4 > β-GP+ZnO > β-GP > AEP+ZnO > AEP > P-free. ZnO NPs promoted the conversion of β-GP, containing C-O-P bonds (0.028-0.041 mg/L), into Pi more efficiently than AEP, which possesses C-P bonds (0.022-0.037 mg/L). Transcriptomics revealed Pi transport/metabolism (phoB (3.99-12.01 fold), phoR (2.20-5.50 fold), ppa (4.49-10.40 fold), and ppk (2.50-5.40 fold)) and phospholipid metabolism (SQD1 (1.85-2.79 fold), SQD2 (2.60-6.53 fold), MGD (2.13-3.21 fold), and DGD (4.08-7.56 fold)) were up-regulated compared to K 2 HPO 4 . 31 P nuclear magnetic resonance spectroscopy identified intracellular P as polyphosphate, orthophosphate, and pyrophosphate. Synchrotron radiation-based X-ray near-edge structure spectroscopy indicated that K 2 HPO 4 and Zn 3 (PO 4 ) 2 in β-GP+ZnO were increased by 8.09% and 7.28% compared to AEP+ZnO, suggesting superior P storage in β-GP+ZnO. Overall, ZnO NPs improved photoinduced electron-hole pair separation and charge separation efficiency and amplified the ·OH and ·O 2 - levels, promoting OP photoconversion into Pi and algae growth.

Primary excitation energy transfer and charge separation in photosystem I (PSI) from the extremophile desert green alga Chlorella ohadii grown in low light were studied using broadband femtosecond pump-probe spectroscopy in the spectral range from 400 to 850 nm and in the time range from 50 fs to 500 ps. Photochemical reactions were induced by the excitation into the blue and red edges of the chlorophyll Qy absorption band and compared with similar processes in PSI from the cyanobacterium Synechocystis  sp. PCC 6803. When PSI from C. ohadii was excited at 660 nm, the processes of energy redistribution in the light-harvesting antenna complex were observed within a time interval of up to 25 ps, while formation of the stable radical ion pair P 700 + A 1 - was kinetically heterogeneous with characteristic times of 25 and 120 ps. When PSI was excited into the red edge of the Qy band at 715 nm, primary charge separation reactions occurred within the time range of 7 ps in half of the complexes. In the remaining complexes, formation of the radical ion pair P 700 + A 1 - was limited by the energy transfer and occurred with a characteristic time of 70 ps. Similar photochemical reactions in PSI from Synechocystis  6803 were significantly faster: upon excitation at 680 nm, formation of the primary radical ion pairs occurred with a time of 3 ps in ~30% complexes. Excitation at 720 nm resulted in kinetically unresolvable ultrafast primary charge separation in 50% complexes, and subsequent formation of P 700 + A 1 - was observed within 25 ps. The photodynamics of PSI from C. ohadii was noticeably similar to the excitation energy transfer and charge separation in PSI from the microalga Chlamydomonas reinhardtii ; however, the dynamics of energy transfer in C. ohadii PSI also included slower components.

This study established and investigated continuous macular pigment (MP) production with a lutein (L):zeaxanthin (Z) ratio of 4-5:1 by an MP-rich Chlorella sp. CN6 mutant strain in a continuous microalgal culture module. Chlorella sp. CN6 was cultured in a four-stage module for 10 days. The microalgal culture volume increased to 200 L in the first stage (6 days). Biomass productivity increased to 0.931 g/L/day with continuous indoor white light irradiation during the second stage (3 days). MP content effectively increased to 8.29 mg/g upon continuous, indoor white light and blue light-emitting diode irradiation in the third stage (1 day), and the microalgal biomass and MP concentrations were 8.88 g/L and 73.6 mg/L in the fourth stage, respectively. Using a two-step MP extraction process, 80 % of the MP was recovered with a high purity of 93 %, and its L:Z ratio was 4-5:1., 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.)

In aquaculture around the world, sulfamonomethoxine (SMM), a long-acting antibiotic that harms microalgae, is widely employed in combination with trimethoprim (TMP), a synergist. However, their combined toxicity to microalgae under long-term exposures at environmentally relevant concentrations remains poorly understood. Therefore, we studied the effects of SMM single-exposures and co-exposures (SMM:TMP=5:1) at concentrations of 5 μg/L and 500 μg/L on Chlorella pyrenoidosa within one aquacultural drainage cycle (15 days). Photosynthetic activity and N assimilating enzyme activities were employed to evaluate microalgal nutrient assimilation. Oxidative stress and flow cytometry analysis for microalgal proliferation and death jointly revealed mechanisms of inhibition and subsequent self-adaptation. Results showed that exposures at 5 μg/L significantly inhibited microalgal nutrient assimilation and induced oxidative stress on day 7, with a recovery to levels comparable to the control by day 15. This self-adaptation and over 95 % removal of antibiotics jointly contributed to promoting microalgal growth and proliferation while reducing membrane-damaged cells. Under 500 μg/L SMM single-exposure, microalgae self-adapted to interferences on nutrient assimilation, maintaining unaffected growth and proliferation. However, over 60 % of SMM remained, leading to sustained oxidative stress and apoptosis. Remarkably, under 500 μg/L SMM-TMP co-exposure, the synergistic toxicity of SMM and TMP significantly impaired microalgal nutrient assimilation, reducing the degradation efficiency of SMM to about 20 %. Consequently, microalgal growth and proliferation were markedly inhibited, with rates of 9.15 % and 17.7 %, respectively, and a 1.36-fold increase in the proportion of cells with damaged membranes was observed. Sustained and severe oxidative stress was identified as the primary cause of these adverse effects. These findings shed light on the potential impacts of antibiotic mixtures at environmental concentrations on microalgae, facilitating responsible evaluation of the ecological risks of antibiotics in aquaculture ponds., 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.)

Lignocellulosic biomass is a pivotal renewable resource in biorefinery process, requiring pretreatment, primarily chemical pretreatment, for effective depolymerization and subsequent transformation. This process yields solid residue for saccharification and lignocellulosic pretreatment wastewater (LPW), which comprises sugars and inhibitors such as phenols and furans. This study explored the microalgal capacity to treat LPW, focusing on two key hydrolysate inhibitors: furfural and vanillin, which impact the growth of six green microalgae. Chlorella sorokiniana exhibited higher tolerance to furfural and vanillin. However, both inhibitors hindered the growth of C. sorokiniana and disrupted algal photosynthetic system, with vanillin displaying superior inhibition. A synergistic inhibitory effect (Q < 0.85) was observed with furfural and vanillin on algal growth. Furfural transformation to low-toxic furfuryl alcohol was rapid, yet the addition of vanillin hindered this process. Vanillin stimulated carbohydrate accumulation, with 50.48 % observed in the 0.1 g/L furfural + 0.1 g/L vanillin group. Additionally, vanillin enhanced the accumulation of C16: 0 and C18: 2, reaching 21.71 % and 40.36 %, respectively, with 0.1 g/L vanillin. This study proposed a microalgae-based detoxification and resource utilization approach for LPW, enhancing the comprehensive utilization of lignocellulosic components. The observed biomass modifications also suggested potential applications for biofuel production, contributing to the evolving landscape of sustainable biorefinery processes., Competing Interests: Declaration of competing interest On behalf of all authors involved in this manuscript entitled “Harmony in Detoxification: Microalgae Unleashing the Potential of Lignocellulosic Pretreatment Wastewater for Resource Utilization”, we declare 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.)

To reduce the high cost of organic carbon sources in waste resource utilization in the cultivation of microalgae, volatile fatty acids (VFAs) derived from activated sludge were used as the sole carbon source to culture Chlorella sorokiniana under the heterotrophic cultivation. The addition of VFAs in the heterotrophic condition enhanced the total nitrogen (TN) and phosphorus (TP) removal of C. sorokiniana, which proved the advantageous microalgae in using VFAs in the heterotrophic culture after screening in the previous study. To discover the possible mechanism of nitrogen and phosphorus adsorption in heterotrophic conditions by microalgae, the effect of different ratios of VFAs (acetic acid (AA): propionic acid (PA): butyric acid (BA)) on the nutrient removal and growth properties of C. sorokiniana was studied. In the 8:1:1 group, the highest efficiency (77.19%) of VFAs assimilation, the highest biomass (0.80 g L -1 ) and lipid content (31.35%) were achieved, with the highest TN and TP removal efficiencies of 97.44 % and 91.02 %, respectively. Moreover, an aerobic denitrifying bacterium, Pseudomonas, was determined to be the dominant genus under this heterotrophic condition. This suggested that besides nitrate uptake and utilization by C. sorokiniana under the heterotrophy, the conduct of the denitrification process was also the main reason for obtaining high nitrogen removal efficiency., 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 and macrophytes are commonly used as human and animal food supplements. We examined the cultivation of the microalgae Chlorella sorokiniana and the duckweed Lemna minor in thermal waters under batch and sequencing batch conditions and we characterized the produced biomass for the presence of essential nutrients as well as for heavy metals and radioisotope content. The highest specific growth rate for the microalgae was observed when 5 or 15 mg/L N were supplemented while the optimal conditions for Lemna minor were observed in the co-presence of 5 mg/L N and 1.7 mg/L P. Lemna minor presented higher concentrations of proteins and lipids comparing to the studied microalgae. Both organisms contained high amounts of lutein (up to 1378 mg/kg for Lemna minor) and chlorophyll (up to 1518 mg/kg for Lemna minor) while β-carotene and tocopherols were found at lower concentrations, not exceeding a few tens of mg/kg. The heavy metal content varied between the two species. Lemna minor accumulated more Cd, Cu, K, Mn, Na, Ni, and Zn whereas Al, Ca and Mg were higher in Chlorella sorokiniana. Both organisms could be a significant source of essential metals but the occasional exceedance of the statutory levels of toxic metals in food products raises concern for potential risk to either humans or animals. Application of gamma-spectroscopy to quantify the effective dose to humans from 228 Ra, 226 Ra and 40 K showed that Chlorella sorokiniana was well under the radiological limits while the collected mass of Lemna minor was too small for radiological measurements with confidence., 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.)

Artemisinin, a novel plant allelochemical, has attracted attention for its potential selective inhibitory effects on algae, yet to be fully explored. This study compares the sensitivity and action targets of Microcystis aeruginosa (M. aeruginosa) and Chlorella pyrenoidosa (C. pyrenoidosa) to artemisinin algaecide (AMA), highlighting their differences. Results indicate that at high concentrations, AMA displaces the natural PQ at the Q B binding site within M. aeruginosa photosynthetic system, impairing the D1 protein repair function. Furthermore, AMA disrupts electron transfer from reduced ferredoxin (Fd) to NADP + by interfering with the iron-sulfur clusters in the ferredoxin-NADP + reductases (FNR) domain of Fd. Moreover, significant reactive oxygen species (ROS) accumulation triggers oxidative stress and interrupts the tricarboxylic acid cycle, hindering energy acquisition. Notably, AMA suppresses arginine synthesis in M. aeruginosa, leading to reduced microcystins (MCs) release. Conversely, C. pyrenoidosa counters ROS accumulation via photosynthesis protection, antioxidant defenses, and by regulating intracellular osmotic pressure, accelerating damaged protein degradation, and effectively repairing DNA for cellular detoxification. Additionally, AMA stimulates the expression of DNA replication-related genes, facilitating cell proliferation. Our finding offer a unique approach for selectively eradicating cyanobacteria while preserving beneficial algae, and shed new light on employing eco-friendly algicides with high specificity., 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 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., 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 low-cost carbon source, acetate, was utilized to feed a linoleic acid-rich Chlorella sorokiniana for microalgal biomass and lipid accumulation. Remarkably high tolerance capability to high acetate dosage up to 30 g/L was observed, with heterotrophy being the preferred trophic mode for algal growth and lipogenesis when supplemented 20 g/L acetate. Transcriptome analysis revealed a marked activation of pathways involved in acetate bioconversion and lipogenesis upon exposure to high-level of acetate. However, the enhancement of photorespiration inhibited photosynthesis, which ultimately led to a decrease in biomass and lipid under mixotrophy. Heterotrophic acetate-feeding generated more superior amino acid profiling of algal biomass and a predominant linoleic acid content (50 %). Heterotrophic repeat fed-batch strategy in 5 L fermenter significantly increased the growth performance and lipid titer, with the highest levels achieved being 23.4 g/L and 7.0 g/L, respectively. This work provides a viable approach for bio-products production through acetate-based heterotrophic algal 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.)

In this work, microalgae cultivation trials were carried out in a membrane bioreactor to investigate fouling when the cultures of Chlorellavulgaris were grown under mixotrophic, heterotrophic, and phototrophic cultivation regimes. The Chlorella cultures were cultivated in wastewater as a source of nutrients that contained a high concentration of ammonium. In mixotrophic cultivation trials, the results showed that the elevated contents of carbohydrates in the soluble microbial product and proteins in extracellular polymeric substances probably initiated membrane fouling. In this case, the highest protein content was also found in extracellular polymeric substances due to the high nitrogen removal rate. Consequently, transmembrane pressure significantly increased compared to the phototrophic and heterotrophic regimes. The data indicated that cake resistance was the main cause of fouling in all cultivations. Higher protein content in the cake layer made the membrane surface more hydrophobic, while carbohydrates had the opposite effect. Compared to a mixotrophic culture, a phototrophic culture had a larger cell size and higher hydrophobicity, leading to less membrane fouling. Based on our previous data, the highest ammonia removal rate was reached in the mixotrophic cultures; nevertheless, membrane fouling appeared to be the fundamental problem., Competing Interests: The authors declare there is no conflict., (© 2024 The Authors This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC BY 4.0), which permits copying, adaptation and redistribution, provided the original work is properly cited (http://creativecommons.org/licenses/by/4.0/).)

The cost and efficiency of an algal-BS treatment system are determined by the specific microalgal species and BS pretreatment method. This study examines the growth of a novel algae Chlorella sp. YSD-2 and the removal of nutrients from the BS using different pretreatment methods, including dilution ratio and sterilization. The highest biomass production (1.84 g L -1 ) was achieved in the 1:2 unsterilized biogas slurry, which was 2.03 times higher than that in the sterilized group, as well as higher lipid productivity (17.29 mg L -1 d -1 ). Nevertheless, the sterilized biogas slurry at a 1:1 dilution ratio exhibited the most notable nutrient-removal efficiency, with COD at 71.97%, TP at 91.32%, and TN at 88.80%. Additionally, the analysis of 16S rRNA sequencing revealed a significant alteration in the indigenous bacterial composition of the biogas slurry by microalgal treatment, with Proteobacteria and Cyanobacteria emerging as the predominant phyla, and unidentified&#95;Cyanobacteria as the primary genus. These findings suggest that Chlorella sp. YSD-2 exhibits favorable tolerance and nutrient-removal capabilities in unsterilized, high-strength biogas slurry, along with high productivity of biomass and lipids. Consequently, these results offer a theoretical foundation for the development of an efficient and economically viable treatment method for algal-BS., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The global energy crisis has spurred a shift from conventional to clean and sustainable energy sources. Biomass derived from microalgae is emerging as an alternative energy source with diverse applications. Despite the numerous advantages of microalgae, large-scale biomass harvesting is not economical and convenient. Self-flocculation is considered an effective phenomenon facilitated by extracting the flocculating substances from microalgae that assist aggregation of algal cells into flocs. A novel cellulose-based bioflocculant has been synthesized from sewage water grown Chlorella sorokiniana and Scenedesmus abundans for harvesting application. The produced bioflocculant amounted to 38.5% and 19.38% of the dry weight of S. abundans and C. sorokiniana, respectively. Analysis via FTIR, XRD, and FESEM-EDX revealed the presence of cellulose hydroxyapatite (HA) in algae-derived cellulose. Harvesting efficiencies of 95.3% and 89.16% were attained for S. abundans and C. sorokiniana, respectively, at a dosage of 0.5 g/L. Furthermore, the bioflocculant was recovered, enabling its reuse with recovery efficiencies of 52% and 10% for S. abundans and C. sorokiniana, respectively. This simple and efficient approach has the potential to replace other harvesting methods, thereby contributing to the economic algal biofuel production., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Excessive proliferation of algae in water depletes dissolved oxygen, resulting in the demise of aquatic life and environmental damage. This study delves into the effectiveness of the dielectric barrier discharge (DBD) plasma activated peracetic acid (PAA) system in deactivating Chlorella. Within 15 min, the algae removal effectiveness reached 89 % under ideal trial conditions. DBD plasma activation of PAA augmented the concentration of reactive species such as ·OH, 1 O 2 , and organic radicals (RO·) in the solution, which are involved in the process of cell inactivation. Reactive oxygen species (ROS) within Chlorella cells continued to rise as a result of treatment-induced damage to the morphological structure and cell membrane of the organism. DNA and chlorophyll-a (Chl-a), were oxidized and destroyed by these invasive active compounds. This study presents an efficient advanced oxidation method to destroy algal cells and adds an alternative strategy for algal control in areas where eutrophication occurs., 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.)

A halotolerant consortium between microalgae and methanotrophic bacteria could effectively remediate in situ CH 4 and CO 2 , particularly using saline wastewater sources. Herein, Methylomicrobium alcaliphilum 20Z was demonstrated to form a mutualistic association with Chlorella sp. HS2 at a salinity level above 3.0%. Co-culture significantly enhanced the growth of both microbes, independent of initial inoculum ratios. Additionally, increased methane provision in enclosed serum bottles led to saturated methane removal. Subsequent analyses suggested nearly an order of magnitude increase in the amount of carbon sequestered in biomass in methane-fed co-cultures, conditions that also maintained a suitable cultural pH suitable for methanotrophic growth. Collectively, these results suggest a robust metabolic coupling between the two microbes and the influence of the factors other than gaseous exchange on the assembled consortium. Therefore, multi-faceted investigations are needed to harness the significant methane removal potential of the identified halotolerant consortium under conditions relevant to real-world operation scenarios., 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.)

Effects of a phosphorus-solubilizing bacteria (PSB) Bacillus megatherium on growth and lipid production of Chlorella sorokiniana were investigated in synthesized swine wastewater with dissolved inorganic phosphorus (DIP), insoluble inorganic phosphorus (IIP), and organic phosphorus (OP). The results showed that the PSB significantly promoted the algal growth in OP and IIP, by 1.10 and 1.78-fold, respectively. The algal lipid accumulation was also greatly triggered, respectively by 4.39, 1.68, and 1.38-fold in DIP, IIP, and OP. Moreover, compared with DIP, OP improved the oxidation stability of algal lipid by increasing the proportion of saturated fatty acids (43.8 % vs 27.9 %), while the PSB tended to adjust it to moderate ranges (30.2-41.6 %). Further, the transcriptome analysis verified the OP and/or PSB-induced up-regulated genes involving photosynthesis, lipid metabolism, signal transduction, etc. This study provided novel insights to enhance microalgae-based nutrient removal combined with biofuel production in practical wastewater, especially with complex forms of phosphorus., 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.)

The nitrogen transformation, enzymatic activity, antioxidant ability and transcriptional response of Chlorella pyrenoidosa (C. pyrenoidosa) treating mariculture wastewater were compared under different light intensities. The microalgal growth, chlorophyll synthesis and nitrogen removal ability of C. pyrenoidosa increased with the light intensity from 3000 to 7000 Lux, whereas they slightly decreased under 9000 and 11,000 Lux. The nitrogen metabolism enzymatic activities displayed obvious differences under different light intensities and affected the nitrogen transformation process. The reactive oxygen species (ROS) production increased with the increase of operational time, whereas it had distinct differences under different light intensities. The changes of antioxidant enzymatic activities were positively correlated with the ROS production. The transcriptional response of C. pyrenoidosa was in accordance with the variation of the photosynthesis, nitrogen assimilation and antioxidant system under different light intensities. This study provides theoretical basis and technical support to select suitable light intensity for algae treating mariculture 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 Ltd. All rights reserved.)

Individual biological water treatment techniques often prove ineffective in removing accumulated high concentrations of nitrogen and phosphorus in the late stages of biofloc aquaculture. To address this issue, we integrated a previously developed autotrophic denitrification and nitrification integrated constructed wetland (ADNI-CW) with a microalgal membrane photobioreactor (MPBR). Under high nitrogen and phosphorus pollution loads in the influent, the standalone ADNI-CW system achieved removal rates of only 24.17 % ± 2.82 % for total nitrogen (TN) and 25.30 % ± 2.59 % for total phosphorus (TP). The optimal conditions for TN and TP degradation and microalgal biomass production in the Chlorella MPBR, determined using response surface methodology, were an inoculum OD 680 of 0.394, light intensity of 161.583 μmol/m 2 /s, and photoperiod of 16.302 h light:7.698 h dark. Under the optimal operating conditions, the integrated ADNI-CW-MPBR system achieved remarkable TN and TP removal rates of 92.63 % ± 2.8 % and 77.46 % ± 8.41 %, respectively, and a substantial microalgal biomass yield of 54.58 ± 6.8 mg/L/day. This accomplishment signifies the successful achievement of efficient nitrogen and phosphorus removal from high-pollution-load marine aquaculture wastewater along with the acquisition of valuable microalgal biomass. A preliminary investigation of the microbial community composition and algal-bacterial interactions in different operational stages of the MPBR system revealed that unclassified&#95;d&#95;&#95;Bacteria, Chlorophyta, and Planctomycetes were predominant phyla. The collaborative relationships between bacteria and Chlorella surpassed competition, ensuring highly efficient nitrogen and phosphorus removal in the MPBR system. This study laid the foundation for the green and sustainable development of the aquaculture industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that may appear to have influenced the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

As one of the most commonly used biocidal cationic surfactants, benzalkonium chlorides (BACs) have been an increasing concern as emerging contaminants. Wastewater has been claimed the main point for BACs to enter into the environment, but to date, it is still largely unknown how the BACs affect the microbes (especially microalgae) in the practical wastewater and how to cost-effectively remove them. In this study, the inhibitory effects of a typical BACs, dodecyl dimethyl benzyl ammonium chloride (DDBAC), on a green microalga Chlorella sp. in oxidation pond wastewater were investigated. The results showed that though a hermetic effect at the first 2 days was observed with the DDBAC at low concentration (<6 mg/L), the algal growth and photosynthesis were significantly inhibited by the DDBAC at all the tested concentrations (3 to 48 mg/L). Fortunately, a new microbial consortium (MC) capable of degrading DDBAC was screened through a gradient domestication method. The MC mainly composed of Wickerhamomyces sp., Purpureocillium sp., and Achromobacter sp., and its maximum removal efficiency and removal rate of DDBAC (48 mg/L) respectively reached 98.1 % and 46.32 mg/L/d. Interestingly, a microbial-microalgal system (MMS) was constructed using the MC and Chlorella sp., and a synergetic effect between the two kinds of microorganisms was proposed: microalga provided oxygen and extracellular polysaccharides as co-metabolic substrates to help the MC to degrade DDBAC, while the MC helped to eliminate the DDBAC-induced inhibition on the alga. Further, by observing the seven kinds of degradation products (mainly including CH 5 O 3 P, C 6 H 5 CH 2 -, and C 8 H 11 N), two possible chemical pathways of the DDBAC degradation were proposed. In addition, the metagenomic sequencing results showed that the main functional genes of the MMS included antibiotic-resistant genes, ABC transporter genes, quorum sensing genes, two-component regulatory system genes, etc. This study provided some theoretical and application findings for the cost-effective pollution prevention of BACs in 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.)

The mitochondria are known to exert significant influence on various aspects of cancer cell physiology. The suppression of mitochondrial function represents a novel avenue for the advancement of anti-cancer pharmaceuticals. The heat shock protein HSP90 functions as a versatile regulator of mitochondrial metabolism in cancer cells, rendering as a promising target for anticancer interventions. In this work, a novel acid polysaccharide named as XQZ3 was extracted from Chlorella pyrenoidosa and purified by DEAE-cellulose and gel-filtration chromatography. The structural characteristic of XQZ3 was evaluated by monosaccharides composition, methylation analysis, TEM, FT-IR, and 2D-NMR. It was found that XQZ3 with a molecular weight of 29.13 kDa was a complex branched polysaccharide with a backbone mainly composed of galactose and mannose. It exhibited good antitumor activity in vitro and in vivo by patient-derived 3D organoid models and patient-derived xenografts models. The mechanistic investigations revealed that XQZ3 specifically interacted with HSP90, impeding the activation of the HSP90/AKT/mTOR signaling cascade. This, in turn, led to the induction of mitochondrial dysfunction, autophagy, and apoptosis, ultimately resulting in the demise of cancer cells due to nutrient deprivation. This study offers a comprehensive theoretical foundation for the advancement of XQZ3, a novel polysaccharide inhibitor targeting HSP90, with potential as an effective therapeutic agent against cancer., Competing Interests: Declaration of competing interest The authors report no declarations of interest. All authors have reviewed the final version of the manuscript and approved it for publication. This original manuscript has neither been formally published nor considering for publication in elsewhere., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The reuse of wastewater after seawater cultivation is critically important. In this study, a phosphorus-supplemented seawater-wastewater cyclic system (PSSWCS) based on Chlorella pyrenoidosa SDEC-35 was developed. With the addition of phosphorus, the algal biomass and the ability to assimilate nitrogen and carbon were improved. At the nitrogen to phosphorus ratio of 20:1, the biomass productivity per mass of nitrogen reached 3.6 g g -1 (N) day -1 in the second cycle. After the third cycle the protein content reached 35.7% of dry mass, and the major metabolic substances in PSSWCS reached the highest content level of 89.5% (35.7% protein, 38.3% lipid, and 15.5% carbohydrate). After the fourth cycle the lipid content maintained at 40.1%. Furthermore, 100.0% recovery of wastewater in PSSWCS increased the nitrogen and carbon absorption to 15.0 and 396.8 g per tonne of seawater. This study achieved seawater-wastewater recycle and produced high-lipid and high-protein algae by phosphorus addition., 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 ubiquity of hexavalent chromium (Cr(VI)) from industrial activities poses a critical environmental threat due to its persistence, toxicity and mutagenic potential. Traditional physico-chemical methods for its removal often entail significant environmental drawbacks. Recent advancements in remediation strategies have emphasized nano and bioremediation techniques as promising avenues for cost-effective and efficient Cr(VI) mitigation. Bioremediation harnesses the capabilities of biological agents like microorganisms, and algae to mitigate heavy metal contamination, while nano-remediation employs nanoparticles for adsorption purposes. Various microorganisms, including E. coli, Byssochlamys sp., Pannonibacter phragmitetus, Bacillus, Aspergillus, Trichoderma, Fusarium, and Chlorella utilize bioreduction, biotransformation, biosorption and bioaccumulation mechanisms to convert Cr(VI) to Cr(III). Their adaptability to different environments and integration with nanomaterials enhance microbial activity, offering eco-friendly solutions. The study provides a brief overview of metabolic pathways involved in Cr(VI) bioreduction facilitated by diverse microbial species. Nitroreductase and chromate reductase enzymes play key roles in nitrogen and chromium removal, with nitroreductase requiring nitrate and NADPH/NADH, while the chromium reductase pathway relies solely on NADPH/NADH. This review investigates the various anthropogenic activities contributing to Cr(VI) emissions and evaluates the efficacy of conventional, nano-remediation, and bioremediation approaches in curbing Cr(VI) concentrations. Additionally, it scrutinizes the mechanisms underlying nano-remediation techniques for a deeper understanding of the remediation process. It identifies research gaps and offers insights into future directions aimed at enhancing the real-time applicability of bioremediation methods for mitigating with Cr(VI) pollution and pave the way for sustainable remediation solutions., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)