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Chlorella vulgaris and Tetraselmis chuii are two microalgae species already marketed because of their richness in high-value and health-beneficial compounds. Previous studies have demonstrated the biological properties of compounds isolated from both microalgae, although data are not yet available on the impact that pre-treatment and gastrointestinal digestion could exert on these properties. The aim of the present study was to analyze the impact of the biomass pre-treatment (freeze/thaw cycles plus ultrasounds) and simulated gastrointestinal digestion in the bioaccessibility and in vitro antioxidant activity (ABTS, ORAC, Q-FRAP, Q-DPPH) of the released digests. The cell wall from microalgae were susceptible to the pre-treatment and the action of saliva and gastric enzymes, releasing bioactive peptides and phenolic compounds that contributed to the potent antioxidant activity of digests through their radical scavenging and iron reduction capacities. Our findings suggest the potential of these microalgae against oxidative stress-associated diseases at both, intestinal and systemic level., 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.)

Arthrospira (Limnospira) maxima (A. maxima) and Chlorella vulgaris (Ch. vulgaris) are among the approved microalgae and cyanobacteria (MaC) in the food industry that are known to be safe for consumption. However, both organisms are controversial regarding their vitamin B 12 content, due to the possible occurrence of pseudo-cobalamin. Concurrently, their nutrition profiles remain understudied. The main purpose of the present study was to identify their nutrition profiles, focusing mainly on vitamin B 12 , amino acids, and micronutrients under iron-induced hormesis (10 mg/L Fe in treated samples). Our findings indicate a higher B 12 content in A. maxima compared to Ch. vulgaris (both control and treated samples). Using liquid chromatography with tandem mass spectrometry (LC-MS/MS), the cyanocobalamin content was determined as 0.42 ± 0.09 μg/g dried weight (DW) in the A. maxima control and 0.55 ± 0.02 μg/g DW in treated A. maxima, resulting in an insignificant difference. In addition, the iron-enriched medium increased the amount of iron in both tested biomasses (p < 0.01). However, a more pronounced (approximately 100×) boost was observed in Ch. vulgaris, indicating a better absorption capacity (control Ch. vulgaris 0.16 ± 0.01 mg/g Fe, treated Ch. vulgaris 15.40 ± 0.34 mg/g Fe). Additionally, Ch. vulgaris also showed a higher micronutrient content. Using both tested microalgae, meeting the sufficient recommended daily mineral allowance for an adult is possible. By combining biomass from A. maxima and Ch. vulgaris in a ratio of 6:1, we can fulfill the recommended daily allowance of vitamin B 12 and iron by consuming 6 tablets/6 g. Importantly, iron hormesis stimulated amino acid composition in both organisms. The profile of amino acids may suggest these biomasses as promising potential nutrition sources., 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.)

Reduced-fat labneh, while offering health benefits, often presents a challenge due to its diminished nutritional profile compared to full-fat varieties. Microalgae, such as Spirulina platensis and Chlorella vulgaris, are increasingly explored for their potential to fortify foods with essential nutrients. This study innovatively investigates the use of these microalgae to enhance the quality of reduced-fat labneh. The effect of incorporating different concentrations of both microalgae was investigated at different concentrations (0.25, 0.5, and 1%) on nutritional profile (including total solids, fat, protein, carbohydrates, essential amino acids, unsaturated fatty acids, pigments, and phenolic compounds), antioxidant activity, texture, sensory attributes, and viability of the starter culture. The findings revealed that 0.25 and 0.5% concentrations of both microalgae positively influenced the sensory characteristics of the labneh and significantly enhanced its nutritional profile. However, a 1% concentration negatively impacted sensory qualities. Chlorella vulgaris enrichment resulted in higher pH values but compromised texture attributes. Importantly, both microalgae varieties enhanced the viability of the starter culture during 21 days of refrigerated storage. The scanning electron microscope images provide visual evidence of the microstructural changes in labneh with varying concentrations of microalgae and over different storage periods. This research establishes the optimal concentrations for individual microalgae enrichment in reduced-fat labneh, offering valuable insights into their potential to improve both nutritional and sensory aspects. However, it's important to mention that while both microalgae have similar effects, they might differ in their specific impacts due to their unique nutritional profiles and physical properties. Therefore, further investigations could explore optimizing a microalgae mixture and its potential application in functional food development., (© 2024. The Author(s).)

Solid-state cultivation is a promising technology for algal biomass production, achieving high productivities without the need for dewatering. However, such systems have suffered from high evaporation, and capital costs. Here is described a hydrogel photobioreactor (hPBR) with the aim of reducing water demand in solid-state cultivations. Two designs are described with "Design A" offering better humidity control overgrowth conditions. A biomass productivity of 2.41gm -2 d -1 , and 2.87gm -2 d -1 when using physically crosslinked poly(vinyl alcohol) (pPVA) and chemically crosslinked PVA (cPVA) respectively were achieved with Chlorella vulgaris with a water demand around 0.44 kg g -1 of biomass. Over the 23 days of growth, the lipid content increased from 18.9 % to 56.6 % and 13.8 % to 43.2 % for pPVA and cPVA respectively, and the chlorophyll content decreased by more than 81 %. However, cell viability stayed high at over 98 % and surface coverage analysis showed good coverage of the gel surface., 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.)

Contributing to the assessment of potential physiological changes in microalgae subjected to different concentrations and types of cyanotoxins, this study investigated the inhibitory effects of cyanotoxins on the growth, density, biomass, and ecotoxicity of Chlorella vulgaris. Chlorella vulgaris was exposed to crude extracts of cyanobacteria producing microcystin-LR (MC-LR), saxitoxin (SXT), anatoxin-a (ATX-A), and cylindrospermopsin (CYN) with initial concentrations of 5.0, 2.05, 0.61, and 1.42 μg.L -1 , respectively. The experiments were conducted under controlled conditions, and monitoring of growth and cell inhibition occurred at 24h, 48h, 72h, and 96h. Chlorophyll-a content and ecotoxicity assessment were conducted with samples collected after 96h of exposure to cyanotoxins. The growth assays of Chlorella vulgaris, with results expressed in terms of average growth rates (doublings/day), indicated the following order for cyanotoxins: SXT (2.03) > CYN (1.66) > MC-LR (1.56) > ATX-A (0.18). This assay revealed the prominent inhibitory potential of ATX-A on Chlorella vulgaris growth compared to the other toxins evaluated. Regarding the inhibition of the photosynthetic process, expressed in terms of the percentage inhibition of Chlorophyll-a, the following order for cyanotoxins was obtained: ATX-A (82%) > MC-LR (76%) > STX (46%) > CYN (16%). These results also indicated that among the cyanotoxins, ATX-A was the most detrimental to the photosynthetic process. However, contrary to the observations in the growth study, SXT proved to be more harmful than CYN in terms of Chlorophyll-a inhibition. Finally, the results of the toxicity assay revealed that only ATX-A and MC-LR exerted a chronic influence on Chlorella vulgaris under the investigated conditions., 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 are increasingly recognized as promising organisms for bioremediation of organic pollutants. This study investigates the potential of enhancing the bioremediation efficiency of pyrene (PYR), a polycyclic aromatic hydrocarbon (PAH), through NaCl induced physiological and biochemical alterations in two microalgae species, Chlorella vulgaris and Scenedesmus acutus. Our findings reveal significant improvement in PYR removal when these microalgae were cultivated in the presence of 0.1% NaCl where PYR removal increased from 54 to 74% for C. vulgaris and from 26 to 75% for S. acutus. However, it was observed that NaCl induced stress had varying effects on the two species. While C. vulgaris exhibited increased PYR removal, it experienced reduced growth and biomass production, as well as lower photosynthetic efficiency when exposed to PYR and PYR + NaCl. In contrast, S. acutus displayed better growth and biomass accumulation under PYR + NaCl conditions, making it a more efficient candidate for enhancing PYR bioremediation in the presence of NaCl. In addition to assessing growth and biochemical content, we also investigated stress biomarkers, such as lipid peroxidation, polyphenol and proline contents. These findings suggest that S. acutus holds promise as an alternative microalgae species for PYR removal in the presence of NaCl, offering potential advantages in terms of bioremediation efficiency and ecological sustainability. This study highlights the importance of understanding the physiological and biochemical responses of microalgae to environmental stressors, which can be harnessed to optimize bioremediation strategies for the removal of organic pollutants like PYR., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Heat stress can greatly challenge growth and meat quality of broiler chickens where research is looking for sustainable ingredients, such as microalgae, that could also alleviate its negative impacts. Thus, in the present study, 576 1-D-old chicks (Ross 308) were housed until commercial slaughtering (42 D) in 36 pens in 2 rooms of a poultry house, according to a full factorial design encompassing 2 room temperatures (standard vs. high), 2 sexes (females vs. males), and 3 dietary treatments, that is, diet C0 (control diet), diet C3, and diet C6 containing 0, 3, and 6%, respectively, of C. vulgaris meal replacing the same quantities of soybean meal. The highest inclusion level of C. vulgaris decreased feed intake (P < 0.001) and body weight (P < 0.0001) compared to the control diet; it increased yellow and red indexes (P < 0.0001) of the breast muscle, besides the proportion of n3 polyunsaturated fatty acids (PUFA) (P = 0.028). Heat stress decreased feed intake (P = 0.001), breast (P = 0.001) and p. major yields (P = 0.036), and increased meat pH (P= 0.008) and cooking losses (P < 0.001), umami (P = 0.021) and brothy flavor (P < 0.001), and the proportion of n3 PUFA rates (P = 0.027), while reducing the contents of several amino acids in the breast meat (P ≤ 0.05). Compared to females, males displayed higher feed intake and growth, and more favorable feed conversion (P < 0.001). Carcass and p. major yields were greater in females (P < 0.001) which also showed a higher occurrence of spaghetti meat compared to males (P < 0.001). In conclusion, C. vulgaris can be used to replace until 3% of soybean meal in diets for broiler chickens without negative implications, while positively affecting breast meat color according to consumers' preferences. However, the microalgae inclusion did not mitigate the negative effects of a chronic heat stress on growth performance nor reduced the occurrence of any myopathies., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Due to the limitations of Chagas disease therapy, microalgae can be promising in the search of new trypanocidal compounds, since these organisms produce bioactive compounds with large pharmaceutical applications, including antiparasitic effects. In this work, trypanocidal activity of aqueous extract of Tetradesmus obliquus and, for the first time, aqueous extract of Chlorella vulgaris, were evaluated against trypomastigote forms of Trypanosoma cruzi . In addition, cytotoxic activity in Vero cells was evaluated. Our results showed that C. vulgaris and T. obliquus present trypanocidal activity (IC 50 = 32.9 µg ml -1 and 36.4 µg ml -1 , respectively), however, C. vulgaris did not present cytotoxic effects in Vero cells (CC 50 > 600 µg ml -1 ) and displayed a higher selectivity against trypomastigotes forms of T. cruzi (SI > 18). Thus, microalgae extracts, such as aqueous extract of C. vulgaris , are promising potential candidates for the development of natural antichagasic drugs.

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

The microalgae Chlorella vulgaris and Tetraselmis chui are valued for their nutrient-rich content, including lipids and polyunsaturated fatty acids (PUFA). However, little is known about how storage and processing affect their lipid quality. This study aimed to assess the impact of domestic storage and cooking practices in dried biomass of C. vulgaris and T. chui . Four conditions were tested: control (newly opened package), light (storage at room temperature and daily light regimen for three weeks), frozen (storage in the freezer at -20 °C for three weeks), and heated (three cycles of 90 min at 100 °C). Lipid extracts were analyzed by GC-MS and LC-MS, and antioxidant activity through DPPH and ABTS radical scavenging assays. Tested storage conditions promoted a decrease in fatty acid content and in diacyl/lyso lipid species ratios of phospholipid (PC/LPC, PE/LPE) and betaine lipids (DGTS/MGTS). Lipid extracts from light treatment showed the lowest antioxidant activity in C. vulgaris (ABTS, IC40: 104.9; DPPH, IC20: 187.9 ± 15.0), while heat affected the antioxidant activity of T. chui (ABTS, IC40: 88.5 ± 2.8; DPPH, IC20 209.4 ± 10.9). These findings underscore the impact of managing storage and processing conditions to optimize the nutritional and functional benefits of C. vulgaris and T. chui in food and feed applications.

Environmental pollution is a big challenge that has been faced by humans in contemporary life. In this context, fossil fuel, cement production, and plastic waste pose a direct threat to the environment and biodiversity. One of the prominent solutions is the use of renewable sources, and different organisms to valorize wastes into green energy and bioplastics such as polylactic acid. Chlorella vulgaris, a microalgae, is a promising candidate to resolve these issues due to its ease of cultivation, fast growth, carbon dioxide uptake, and oxygen production during its growth on wastewater along with biofuels, and other productions. Thus, in this article, we focused on the potential of Chlorella vulgaris to be used in wastewater treatment, biohydrogen, biocement, biopolymer, food additives, and preservation, biodiesel which is seen to be the most promising for industrial scale, and related biorefineries with the most recent applications with a brief review of Chlorella and polylactic acid market size to realize the technical/nontechnical reasons behind the cost and obstacles that hinder the industrial production for the mentioned applications. We believe that our findings are important for those who are interested in scientific/financial research about microalgae., (© 2024 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.)

The green microalga Chlorella vulgaris was used as a test organism during this study for evaluation of the impact of different heavy metal stress, Mn 2+ , Co 2+ , and Zn 2+ , on enhancing the biodiesel production. The algal cultures were grown for 13 days under heavy metal stress after which were subjected to estimation of growth, some primary metabolites, lipid, and fatty acid profiles. The maximum lipid accumulation (283.30 mg/g CDW) was recorded in the algal culture treated with 3 µM cobalt nitrate. Application of 2 mM manganese chloride; 1, 2, and 3 μM cobalt nitrate; and 0.2, 0.4, and 0.6 mM zinc sulfate caused highly significant increases in the lipid contents amounting to 183.8, 191.4, 230.6, 283.3, 176.3, 226.0, and 212.1 mg/g CDW, respectively, in comparison to control (153.4 mg/g CDW). The maximum proportion of saturated fatty acids (SFA) (64.44%) was noted in the culture treated with 6 mM MnCl 2 due to the existence of palmitic acid (C16:0), stearic acid (C18:0), and pentadecylic acid (C15:0) which are represented by 53.59%, 5.96%, and 1.37%, respectively, of the total FAs. Relative increase in energy compound (REEC) showed that 1, 2, and 3 µM Co 2+ lead to the highest stimulation in lipid and carbohydrate contents to 0.207, 0.352, and 0.329 × 10 3 %, respectively. Empirical formulas were used for the assessment of biodiesel fuel properties based on FAME composition. The estimated properties met the prescribed international standard criteria., (© 2024. The Author(s).)

Bioremediation became a promising technology to resolve arsenic (As) contamination in aquatic environment. Since monoculture such as microalgae or bacteria was sensitive to environmental disturbance and vulnerable to contamination, green microalgae Chlorella vulgaris and arsenite (As(III)) - oxidizing bacteria Pseudomonas sp. SMS11 were co-cultured to construct algal-bacterial consortia in the current study. The effects of algae-bacteria (A:B) ratio and exposure As(III) concentration on algal growth, As speciation and metabolomic profile were investigated. Algal growth arrested when treated with 100 mg/L As(III) without the co-cultured bacteria. By contrast, co-cultured with strain SMS11 significantly enhanced As tolerance in C. vulgaris especially with A:B ratio of 1:10. All the As(III) in culture media of the consortia were oxidized into As(V) on day 7. Methylation of As was observed on day 14. Over 1% and 0.5% of total As were converted into dimethylarsinic acid (DMA) after 21 days cultivation when the initial concentrations of As(III) were 1 and 10 mg/L, respectively. Metabolomic analysis was further performed to reveal the response of consortia metabolites to external As(III). The enriched metabolomic pathways were associated with carbohydrate, amino acid and energy metabolisms. Tricarboxylic acid cycle and glyoxylate and dicarboxylate metabolism were upregulated under As stress due to their biological functions on alleviating oxidative stress and protecting cells. Both carbohydrate and amino acid metabolisms provided precursors and potential substrates for energy production and cell protection under abiotic stress. Alterations of the pathways relevant to carbohydrate or amino acid metabolism were triggered by energy requirement., 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.)

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., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Zibarev et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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 (AgNO 3 ). 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., (© 2024 Wiley Periodicals LLC.)

This study investigates nutrient recovery from synthetic municipal wastewater using co-immobilized cultures of Chlorella vulgaris TISTR 8580 (CV) and plant growth-promoting bacteria, Bacillus subtilis TISTR 1415 (BS) as living biofilters for a subsequent biofertilizer activity. The optimal condition for nutrient recovery was at the 1:1 ratio of CV/BS using mixed guar gum/carrageenan (GG/CG) binders. After 7-day wastewater treatment, the living biofilters removed 86.7 ± 0.5% of ammonium and 99.3 ± 0.3% of phosphates and were tested subsequently as biofertilizers for 20 days to grow selected plants. The highest optimal biomass and chlorophyll a content was 2 ± 0.3 g (CV/BS 3:1) and 12.4 ± 0.7 µg/g (CV/BS 1:1) from cucumber respectively, however, the close-to-neutral pH (8.0 ± 0.3) was observed from sunflower using CV/BS 1:1 living biofilters. Conclusively, the designed living biofilters exhibit the potential to recover nutrients from wastewater and be used as biofertilizers for circular agriculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Municipal wastewater is ubiquitously laden with myriad pollutants discharged primarily from a combination of domestic and industrial activities. These heterogeneous pollutants are threating the natural environments when the traditional activated sludge system fails sporadically to reduce the pollutants' toxicities. Besides, the activated sludge system is very energy intensive, bringing conundrums for decarbonization. This research endeavoured to employ Chlorella vulgaris sp. In converting pollutants from municipal wastewater into hydrogen via alternate light and dark fermentative process. The microalgae in attached form onto 1 cm 3 of polyurethane foam cubes were adopted in optimizing light intensity and photoperiod during the light exposure duration. The highest hydrogen production was recorded at 52 mL amidst the synergistic light intensity and photoperiod of 200 μmolm -2 s -1 and 12:12 h (light:dark h), respectively. At this lighting condition, the removals of chemical oxygen demand (COD) and ammoniacal nitrogen were both achieved at about 80%. The sustainability of microalgal fermentative performances was verified in recyclability study using similar immobilization support material. There were negligible diminishments of hydrogen production as well as both COD and ammoniacal nitrogen removals after five cycles, heralding inconsequential microalgal cells' washout from the polyurethane support when replacing the municipal wastewater medium at each cycle. The collected dataset was finally modelled into enhanced Monod equation aided by Python software tool of machine learning. The derived model was capable to predict the performances of microalgae to execute the fermentative process in producing hydrogen while subsisting municipal wastewater at arbitrary photoperiod. The enhanced model had a best fitting of R 2 of 0.9857 as validated using an independent dataset. Concisely, the outcomes had contributed towards the advancement of municipal wastewater treatment via microalgal fermentative process in producing green hydrogen as a clean energy source to decarbonize the wastewater treatment facilities., 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.)

Biomass can be used as an energy source to thermochemical conversion processes to biocrude production. However, the supply and dependence on only one biomass for biocrude production can be an obstacle due to its seasonality, availability, and logistics costs. In this way, biomass waste and agroindustrial residues can be mixture and used as feedstock to the hydrothermal co-liquefaction (co-HTL) process as an alternative to obtaining biocrude. In this sense, the present paper analyzed the biocrude yield influence of the co-HTL from a quaternary unprecedented blend of different biomasses, such as sugarcane bagasse, brewer's spent grain (BSG), sludge from a paper recycling mill (PRM), and microalgae (Chlorella vulgaris). In this way, a simplex lattice design was employed and co-HTL experiments were carried out in a 2000 mL high-pressure stirred autoclave reactor under 275 °C for 60 min, considering 15% of feedstock/water ratio. Significant effects in each feedstock and their blends were analyzed aiming to increase biocrude and biochar yield. It was found that the addition of microalgae is only significant when considered more than 50% into the blend with BSG and PRM sludge to increase biocrude yield., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Mateus de Souza Amaral reports financial support was provided by Minas Gerais StateFoundation of Support to the Research. Sara Cangussu Bassoli reports financial support was provided by Coordination of Higher Education Personnel Improvement., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

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

Pulsed electric field (PEF) processing has emerged as an alternative to thermal pasteurization for the shelf-life extension of heat-sensitive liquids at industrial scale. It offers the advantage of minimal alteration in physicochemical characteristics and functional properties. In this study, a pilot-scale continuous PEF processing (T outlet  < 55 °C) was applied to microalgae Chlorella vulgaris (Cv) suspensions (pH = 6.5), which was proposed as a functional ingredient for plant-based foods. Cv suspensions were inoculated with three distinct food spoilage microorganisms (Pseudomonas guariconensis, Enterobacter soli and Lactococcus lactis), isolated from the Cv biomass. PEF treatments were applied with varying electric field strength E el of 16 to 28 kV/cm, pulse repetition rate f of 100 to 140 Hz, with a pulse width τ of 20 μs and an inlet product temperature T in of 30 °C. The aim was to evaluate the PEF-induced microbial reduction and monitor the microbial outgrowth during a 10-day cold storage period (10 °C). Maximum inactivation of 4.1, 3.7 and 3.6 logs was achieved (28 kV/cm and 120 Hz) for the investigated isolates, respectively. Under these conditions, the critical electric field strengths E crit , above which inactivation was observed, ranged from 22.6 to 24.6 kV/cm. Moreover, repeated PEF treatment resulted in similar inactivation efficiency, indicating its potential to enhance shelf-life further., 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 Author(s). Published by Elsevier Ltd.. All rights reserved.)

The advent of deep learning algorithms for protein folding opened a new era in the ability of predicting and optimizing the function of proteins once the sequence is known. The task is more intricate when cofactors like metal ions or small ligands are essential to functioning. In this case, the combined use of traditional simulation methods based on interatomic force fields and deep learning predictions is mandatory. We use the example of [FeFe] hydrogenases, enzymes of unicellular algae promising for biotechnology applications to illustrate this situation. [FeFe] hydrogenase is an iron-sulfur protein that catalyzes the chemical reduction of protons dissolved in liquid water into molecular hydrogen as a gas. Hydrogen production efficiency and cell sensitivity to dioxygen are important parameters to optimize the industrial applications of biological hydrogen production. Both parameters are related to the organization of iron-sulfur clusters within protein domains. In this work, we propose possible three-dimensional structures of Chlorella vulgaris 211/11P [FeFe] hydrogenase, the sequence of which was extracted from the recently published genome of the given strain. Initial structural models are built using: (i) the deep learning algorithm AlphaFold; (ii) the homology modeling server SwissModel; (iii) a manual construction based on the best known bacterial crystal structure. Missing iron-sulfur clusters are included and microsecond-long molecular dynamics of initial structures embedded into the water solution environment were performed. Multiple-walkers metadynamics was also used to enhance the sampling of structures encompassing both functional and non-functional organizations of iron-sulfur clusters. The resulting structural model provided by deep learning is consistent with functional [FeFe] hydrogenase characterized by peculiar interactions between cofactors and the protein matrix.

The study investigates the potential of utilizing banana trunk-derived porous activated biochar enriched with SO 3 H- as a catalyst for eco-friendly biodiesel production from the microalga Chlorella vulgaris. An extensive analysis, employing advanced techniques such as XRD, FTIR, TGA, XPS, NH 3 -TPD, BET, SEM-EDX, and TEM, was conducted to elucidate the physicochemical properties of BT-SO 3 H catalysts. The synthesized catalyst demonstrated its efficiency in converting the total lipids of Chlorella vulgaris into biodiesel, with varying concentrations of 3%, 5%, and 7%. Notably, using a 5% BT-SO 3 H concentration resulted in remarkably higher biodiesel production about 58.29%. Additionally, the fatty acid profile of C. vulgaris biodiesel indicated that C16:0 was the predominant fatty acid at 24.31%, followed by C18:1 (19.68%), C18:3 (11.45%), and C16:1 (7.56%). Furthermore, the biodiesel produced via 5% BT-SO 3 H was estimated to have higher levels of saturated fatty acids (SFAs) at 34.28%, monounsaturated fatty acids (MUFAs) at 30.70%, and polyunsaturated fatty acids (PUFAs) at 24.24%. These findings highlight the promising potential of BT-SO 3 H catalysts for efficient and environmentally friendly biodiesel production from microalgal species., 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 Elsevier Inc. All rights reserved.)

Aquaculture is a controlled aquatic farming sector and one of the most important human food sources. Fish farming is one of the predominant, fast-growing sectors that supply seafood products worldwide. Along with its benefits, aquaculture practices can discharge large quantities of nutrients into the environment through non-treated or poorly treated wastewater. This study aims to understand the nutrient composition of fish wastewater and the use of indigenous bacteria, cyanobacteria, and microalgae as an alternative biological treatment method. Wastewater samples from a local fish farming facility were collected and treated using six different species of cyanobacteria and microalgae include Chroococcus minutus, Porphyridium cruentum, Chlorella vulgaris, Microcystis aeruginosa, Chlamydomonas reinhardtii, and Fischerella muscicola. All the samples were incubated for 21 days, and the following parameters were measured weekly: Chemical oxygen demand (COD), phosphate, total dissolved nitrogen, and dissolved inorganic nitrogen. In addition, dissolved organic nitrogen (DON), bioavailable DON (ABDON), and biodegradable DON (BDON) were calculated from the mass-balance equations. Colorimetric and digestive methods were used for the parameter measurements. The results showed that C. reinhardtii reduced the soluble COD concentration by 74.6 %, DON by 94.3 %, and phosphorous by more than 99 %. Moreover, M. aeruginosa, and C. minutus significantly reduced inorganic nitrogen species (>99 %). This alternative fish wastewater treatment method was explored to gain insight into fish wastewater nutrient composition and to create a sustainable alternative to conventional fish wastewater treatment methods., 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.)

Human urine is rich in nitrogen and phosphorus, and the presence of these elements in wastewater significantly disrupts the biogeochemical cycle. Meanwhile, green algal biomass cultivation is unfeasible without these nutrients. Hence, the present study integrates wastewater treatment and algae cultivation to extract biodiesel and improve its performance through fuel modification. Chlorella vulgaris algae was cultivated in different dilution ratios of water and urine, and the nutrient removal rate was analyzed. Chlorella vulgaris algae biodiesel (CAB) was derived through Bligh and Dyer's method followed by transesterification, and its functional and elemental groups were analyzed. The various volume concentrations of CAB were blended with regular diesel fuel (RDF), and 10% water was added to a 30% CAB blended RDF to evaluate the combustion performance and environmental impacts. The results of the experiments demonstrated that the algae cultivation effectively removed the wastewater nutrients. The functional and elemental groups of CAB are identical to those of RDF. The engine characteristics of test fuels report that the CAB-blend RDF fuel mixtures generate low carbon footprints, whereas negative impacts have been drawn for performance metrics and oxides of nitrogen emissions. The water-emulsified fuel outweighed the unfavorable effects and promoted more efficient and cleaner combustion., 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.)

Calcium hypochlorite (Ca(ClO) 2 ), which can be stably stored in powder form for a long period, is widely used as a disinfectant in various fields. A new sterilization process was developed in the present study, where a microalgal medium was sterilized using 0.02% Ca(ClO) 2 , followed by complete neutralization of the Ca(ClO) 2 within 8 h through catalytic reaction of an MnCl 2 -Na 2 EDTA complex and a synergistic effect of glucose. When comparing the growth of Chlorella vulgaris in the autoclaved medium, a 2.65 times greater maximum cell growth was observed in cells grown in the medium prepared by treatment of Ca(ClO) 2 . This result indicates that denaturation of the medium by heat can hinder the growth of some microorganisms. In the case of cultivation of Euglena gracilis, successful culture growth was achieved without growth inhibition or contamination on a medium prepared in the same manner., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study assessed the effect of gibberellins (GAs) concentrations on antibiotic and nutrient removal using diverse microalgal-bacterial-fungal consortia. Five systems (Chlorella vulgaris, T1; C. vulgaris + S395-2 + Clonostachys rosea, T2; C. vulgaris + S395-2 + Ganoderma lucidum, T3; C. vulgaris + S395-2 + Pleurotus pulmonarius, T4; and C. vulgaris + S395-2, T5) were established, and optimal conditions and effective symbiosis were applied to improve antibiotic and nutrient removal. Consortium growth was T2 > T3 > T5 > T4 > T1, while GA impact ranked 50 mg L -1  > 20 mg L -1  > 80 mg L -1  > 0 mg L -1 . After 7 days at 50 mg L -1 GAs, total nitrogen (TN), NH 4 -N, NO 3 -N, and total phosphorous (TP) removal reached 85.97 %, 78.08 %, 86.59 %, and 94.39 %, respectively. Florfenicol, oxytetracycline hydrochloride, ofloxacin, and sulfamethoxazole removal efficiencies were 67.77 %, 98.29 %, 90.47 %, and 94.92 %, respectively. These findings highlight GAs' significant role in enhancing antibiotic and nutrient removal., 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.)

Recently, hybrid systems, such as those incorporating high-rate algal ponds (HRAPs) and biofilm reactors (BRs), have shown promise in treating domestic wastewater while cultivating microalgae. In this context, the objective of the present study was to determine an improved scraping frequency to maximize microalgae biomass productivity in a mix of industrial (fruit-based juice production) and domestic wastewater. The mix was set to balance the carbon/nitrogen ratio. The scraping strategy involved maintaining 1 cm wide stripes to retain an inoculum in the reactor. Three scraping frequencies (2, 4, and 6 days) were evaluated. The findings indicate that a scraping frequency of each 2 days provided the highest biomass productivity (18.75 g total volatile solids m -2 d -1 ). The species' behavior varied with frequency: Chlorella vulgaris was abundant at 6-day intervals, whereas Tetradesmus obliquus favored shorter intervals. Biomass from more frequent scraping demonstrated a higher lipid content (15.45%). Extrapolymeric substance production was also highest at the 2-day frequency. Concerning wastewater treatment, the system removed 93% of dissolved organic carbon and ∼100% of ammoniacal nitrogen. Combining industrial and domestic wastewater sources to balance the carbon/nitrogen ratio enhanced treatment efficiency and biomass yield. This study highlights the potential of adjusting scraping frequencies in hybrid systems for improved wastewater treatment and microalgae 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.)

This study presents the catalytic pyrolysis of microalgae, Chlorella vulgaris (C. vulgaris), using pure CH 4 and H 2 -rich gas evolved from CH 4 decomposition on three different HZSM-5 catalysts loaded with Zn, Ga, and Pt, aimed specifically at producing high-value mono-aromatics such as benzene, toluene, ethylbenzene, and xylene (BTEX). In comparison with that for the typical inert N 2 environment, a pure CH 4 environment increased the bio-oil yield from 32.4 wt% to 37.4 wt% probably due to hydrogen and methyl radical insertion in the bio-oil components. Furthermore, the addition of bimetals further increased bio-oil yield. For example, ZnPtHZ led to a bio-oil yield of 47.7 wt% in pure CH 4 . ZnGaHZ resulted in the maximum BTEX yield (6.68 wt%), which could be explained by CH 4 activation, co-aromatization, and hydrodeoxygenation. The BTEX yield could be further increased to 7.62 wt% when pyrolysis was conducted in H 2 -rich gas evolved from CH 4 decomposition over ZnGaHZ, as rates of aromatization and hydrodeoxygenation were relatively high under this condition. This study experimentally validated that the combination of ZnGaHZ and CH 4 decomposition synergistically increases BTEX production using C. vulgaris., 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 research explicitly investigates the utilization of Chlorella Vulgaris sp. microalgae as a renewable source for lipid production, focusing on its application in bioplastic manufacturing. This study employed the supercritical fluid extraction technique employing supercritical CO 2 (sCO 2 ) as a green technology to selectively extract and produce PHA's precursor utilizing CO 2 solvent as a cleaner solvent compared to conventional extraction method. The study assessed the effects of three extraction parameters, namely temperature (40-60 °C), pressure (15-35 MPa), and solvent flow rate (4-8 ml/min). The pressure, flowrate, and temperature were found to be the most significant parameters affecting the sCO 2 extraction. Through Taguchi optimization, the optimal parameters were determined as 60 °C, 35 MPa, and 4 ml/min with the highest lipid yield of 46.74 wt%; above-average findings were reported. Furthermore, the pretreatment process involved significant effects such as crumpled and exhaustive structure, facilitating the efficient extraction of total lipids from the microalgae matrix. This study investigated the microstructure of microalgae biomatrix before and after extraction using scanning electron microscopy (SEM) and thermogravimetric analysis (TGA). Fourier-transform infrared spectroscopy (FTIR) was utilized to assess the potential of the extracted material as a precursor for biodegradable plastic production, with a focus on reduced heavy metal content through inductively coupled plasma-optical emission spectrometry (ICP-OES) analysis. The lipid extracted from Chlorella Vulgaris sp. microalgae was analysed using gas chromatography-mass spectrometry (GC-MS), identifying key constituents, including oleic acid (C 18 H 34 O 2 ), n-Hexadecanoic acid (C 16 H 32 O 2 ), and octadecanoic acid (C 18 H 36 O 2 ), essential for polyhydroxyalkanoate (PHA) formation., 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 utilization of microalgae for both removing phthalate esters (PAEs) from wastewater and producing bioenergy has become a popular research topic. However, there is a lack of studies comparing the effectiveness of different types of microalgae in removing these harmful compounds. Therefore, the present study aimed to evaluate and compare the efficiency of various processes, such as hydrolysis, photolysis, adsorption, and biodegradation, in removing diisobutyl phthalate (DiBP) using six different species of microalgae. The study indicated that the average removal efficiency of DiBP (initial concentrations of 5, 0.5, and 0.05 mg L -1 ) by all six microalgae (initial cell density of 1 × 10 6 cells mL -1 ) was in the order of Scenedesmus obliquus (95.39 %) > Chlorella vulgaris (94.78 %) > Chroococcus sp. (91.16 %) > Cyclotella sp. (89.32 %) > Nitzschia sp. (88.38 %) > Nostoc sp. (84.33 %). The results of both hydrolysis and photolysis experiments revealed that the removal of DiBP had minimal impact, with respective removal efficiencies of only 0.89 % and 1.82 %. The adsorption efficiency of all six microalgae decreased significantly with increasing initial DiBP concentrations, while the biodegradation efficiency was elevated. Chlorella vulgaris and Chroococcus sp. demonstrated the highest adsorption and biodegradation efficiencies among the microalgae tested. Scenedesmus obliquus was chosen for the analysis of the degradation products of DiBP due to its exceptional ability to remove DiBP. The analysis yielded valuable results, identifying monoisobutyl phthalate (MiBP), phthalic acid (PA), and salicylic acid (SA) as the possible degradation products of DiBP. The possible degradation pathways mainly included dealkylation, the addition of hydroxyl groups, and decarboxylation. This study lays a theoretical foundation for the elimination of PAEs in the 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 © 2023 Elsevier B.V. All rights reserved.)

Wine production represents one of the most important agro-industrial sectors in Italy. Wine lees are the most significant waste in the winery industry and have high disposal and storage costs and few applications within the circular economy. In this study, anaerobic digestion and a microalgae coupled process was studied in order to treat wine lees and waste activated sludge produced within the same facility, with the aim of producing energy and valuable microalgae biomass that could be processed to recover biofuel or biostimulant. Chlorella vulgaris was cultivated on liquid digestate in a semi-continuous system without biomass recirculation. The best growth and phytoremediation performance were achieved applying a hydraulic retention time (HRT) of 20 days with a stable dry weight, lipid and protein storage of 1.85 ± 0.02 g l -1 , 33.48 ± 7.54 % and 57.85 ± 10.14 % respectively. Lipid characterization highlighted the potential use in high quality biodiesel production, according to EN14214 (<12 % v/v linolenic acid). The microalgae reactor's liquid output showed high removal of ammonia (95.72 ± 2.10 %), but low organic soluble matter reduction. Further semi-continuous process optimization was carried out by increasing the time between digestate feeding and biomass recovery at HRT 10. These operative changes avoided biomass wash-out and provided a stable phytoremediation of the digestate with 84.58 ± 4.02 % ammonia removal, 33.01 ± 1.44 % sCOD removal, 38.06 ± 2.65 % of polyphenols removal., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Paolina Scarponi has patent #102022000011675 licensed to Licensee.]., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The comprehensive evaluation and validation of mathematical models for microalgal growth dynamics are essential for improving cultivation efficiency and optimising photobioreactor design. A considerable gap in comprehending the relation between microalgal growth, light intensity and biomass concentration arises since many studies focus solely on associating one of these factors. This paper compares microalgal growth kinetic models, specifically focusing on the combined impact of light intensity and biomass concentration. Considering a dataset (experimental results and literature values) concerning Chlorella vulgaris, nine kinetic models were assessed. Bannister and Grima models presented the best fitting performance to experimental data (RMSE ≤ 0.050 d -1 ; R 2 ≥0.804; d 2 ≥0.943). Cultivation conditions conducting photoinhibition were identified in some kinetic models. After testing these models on independent datasets, Bannister and Grima models presented superior predictive performance (RMSE = 0.022-0.023 d -1 ; R 2  = 0.878-0.884; d 2 : 0.976-0.975). The models provide valuable tools for predicting microalgal growth and optimising operational parameters, reducing the need for time-consuming and costly experiments., 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 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Androstenedione (ADSD) is one of the widely detected androgens in diverse aquatic environments. However, there were few reports on the molecular mechanism of Chlorella vulgaris exposure to ADSD. In our previous research, we have investigated the genes associated with chlorophyll metabolism in Chlorella vulgaris response to ADSD. In this study, we focus on continuously up-regulated genes to explore the mechanism underlying Chlorella vulgaris resistance to ADSD toxicity. Chlorella vulgaris was exposed to ADSD with five concentration gradients. The continuously up-regulated genes were enriched by Series Test of Cluster (STC) analysis and verified by qRT-PCR. Microalgae Super Oxidase Dimutase (SOD) and Microalgae Malonic dialdehyde (MDA), two indicators of oxidative stress, were determined by ELISA after exposure to ADSD. The results showed that ADSD can stimulate the production of extracellular polymeric substances (EPS) and lead to enlargement in the cell body of Chlorella vulgaris. In addition, steroid biosynthesis and oxidoreductase activity processes were consistently up-regulated upon exposure to ADSD. In conclusion, our study highlighted the crucial role of phenotypic modification, hormone synthesis, and redox mechanisms in protecting Chlorella vulgaris cells from the harmful effects of ADSD contamination., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

This study explores diverse cultivation modes for Chlorella vulgaris within a biorefinery at pilot scale that produces both biodiesel by transesterification of waste frying oils and syngas by gasification of organic wood waste. Given microalgae's comparatively modest biofuel yield relative to principal biorefinery products, the microalgae cultivation process is designed on the biofuels production rates. Liquid and gaseous waste streams are recycled inside the biorefinery: crude glycerol is mixed with wood to enhance the quality of syngas, wastewater is fed to microalgae so as flue gas. Also, the oil extracted from microalgae contributes to produce biodiesel and the waste cells are gasified. Considering that the optimal fit for each cultivation mode varies with the shape of the reactor, we propose a modular approach to assemble them in batteries of tubular, bubble flow, and airlift reactors, and present an operating design criterion that can fulfill the mass balance of the plant by adding/transforming the number of units inside the different batteries. Methods to adjust the operating conditions and control the operating parameters are also discussed. The designed configurations were operated recycling nominal waste streams of about 30 L d -1 of wastewater and 90 Nm 3 h -1 of flue gas. Results confirm that the most advantageous one, in terms of volume per recycled waste streams, is a battery of 16 airlift reactors, operating in mixotrophic mode, with growing rate of 0.427 d -1 , yield of 3.06, glycerol conversion 39 %, CO 2 removal 64 % of inlet 6-10 %(mol) concentration. The same nominal waste streams can also be managed by 40 tubular reactors in almost heterotrophic conditions coupled with 12 bubble columns in autotrophic conditions; working respectively at growing rates of 0.395 d -1 and 0.362 d -1 and yields of 2.94 and 2.84. The battery of tubular reactors reached a glycerol conversion of 45 % and the array of bubble columns removed about 51 % of inlet 12-20 %(mol) CO 2 concentration. A complete comparison is reported also in terms of dimensionless numbers and pumping/mixing requirements., 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 The Authors. Published by Elsevier Inc. All rights reserved.)

To develop and characterize novel antibiotics removal biomaterial technology, we constructed three different bacteria-microalgae-fungi consortiums containing Chlorella vulgaris (C. vulgaris), endophytic bacterium, Clonostachys rosea (C. rosea), Ganoderma lucidum, and Pleurotus pulmonarius. The results showed that under treatment with 50 mg/L of gibberellins (GAs), the three bacteria-microalgae-fungi symbionts had maximal growth rates (0.317 ± 0.030 d -1 ) and the highest removal efficiency for seven different antibiotics. Among them, C. vulgaris-endophytic bacterium-C. rosea symbiont had the best performance, with antibiotics removal efficiencies of 96.0 ± 1.4 %, 91.1 ± 7.9 %, 48.7 ± 5.1 %, 34.6 ± 2.9 %, 61.0 ± 5.5 %, 63.7 ± 5.6 %, and 54.3 ± 4.9 % for tetracycline hydrochloride, oxytetracycline hydrochloride, ciprofloxacin, norfloxacin, sulfadiazine, sulfamethazine, and sulfamethoxazole, respectively. Overall, the present study demonstrates that 50 mg/L GAs enhances biomass production and antibiotics removal efficiency of bacteria-microalgae-fungi symbionts, providing a framework for future antibiotics-containing wastewater treatment using three-phase symbionts., 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 Elsevier Ltd. All rights reserved.)

Microalgae are considered a promising source for obtaining natural compounds with strong antioxidant activity. Despite the great progress made in this field, there is still need for further studies applying simple and cost-effective modifications to reveal their full potential and enhance antioxidant properties. Arthrospira platensis and Chlorella vulgaris are some of the most common cells studied for this purpose. In this study, it was aimed to develop a bioprocess for the enhancement of antioxidant properties of these two microalgae by evaluating the effect of different culture conditions. With this aim, the impacts of light intensity/reactive oxygen species and nitrogen sources/reactive oxygen species were evaluated for the A. platensis and C. vulgaris cells, respectively. Results showed that the antioxidant potential of A. platensis was found to be correlated with the phycocyanin and total phenolic content of cells, and 80 µmol photons m -2  s -1 light intensity induced antioxidant activity in a two-step cultivation mode. For C. vulgaris cells, maximum antioxidant activities of 68.10 ± 1.51% and 75.68 ± 0.66% were obtained in cultures with NH 4 Cl (0.016% (w/v)) for DPPH and ABTS assays, respectively. The applied oxidative stress factors exhibited different effects on the antioxidant activities of the cells because of their cellular morphologies and changing mechanisms of reactive oxygen species. These outcomes show the potential of applied modifications on cells and suggest a promising route to enhance antioxidant activities of microalgae for further research., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Recent advancement in biophotovoltaic systems using microalgae, coupled with biorefinery approach, would improve economy-feasibility in production. The major concern is its commercial strength in terms of scalability, strain selection and extraction procedure cost. It must compete with conventional feedstocks such as fossil fuels. This project proposes to enhance the economic feasibility of microalgae-based biorefinery by evaluating their performance for bio-electricity, bio-diesel and carotenoids production in a single cycle. The first part of the study was to construct and select a Bio-bottle Voltaic (BBV) device that would allow microalgae to grow and produce bioproducts, as well as generate the maximum current output reading derived from the microalgae's photosynthesis process. The second phase consisted of a 25-day investigation into the biorefinery performance of six different microalgal species in producing bio-electricity, bio-diesel and carotenoid in a prototype BBV device. The prototype BBV device with aluminium foil and pencil lead as its anode and cathode produced the highest carotenoid and biodiesel component production from the two microalgae tested, according to the results of the first phase of the experiment. In the second portion of the study, Scenedesmus dimorphus and Chlorella vulgaris were identified as the two microalgae most capable of maintaining their growth throughout the experiment. The maximum current reading observed for C. vulgaris was 653 mV. High Performance Liquid Chromatography analysis showed four major carotenoid compounds found which were Neoxanthin, Cantaxanthin, Astaxanthin and 9-cis antheraxanthin, and the highest carotenoid producer was C. vulgaris which recorded at 1.73 μg/mL. C. vulgaris recorded as the most alkanes producer with 22 compounds detected and Heptacosane and Heneicosane as the two major biodiesel compounds found in the extracts. Evaluation of C. vulgaris data showed that it has enormous potential for microalgal biorefinery candidates. Further ongoing research and development efforts for C. vulgaris will improve the economic viability of microalgae-based industries and reduce reliance on depleted fossil fuels., 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 Elsevier Ltd. All rights reserved.)

Research on microalgae has surged due to its diverse biotechnological applications and capacity for accumulating bioactive compounds. Despite considerable advancements, microalgal cultivation remains costly, prompting efforts to reduce expenses while enhancing productivity. This study proposes a cost-effective approach through the coculture of microalgae and bacteria, exploiting mutualistic interactions. An engineered consortium of Chlorella vulgaris and Stutzerimonas stutzeri strain J3BG demonstrated biofilm-like arrangements, indicative of direct cell-to-cell interactions and metabolite exchange. Strain J3BG's enzymatic characterization revealed amylase, lipase, and protease production, sustaining mutual growth. Employing Response Surface Methodology (RSM), Artificial Neural Network (ANN), and Genetic Algorithm (GA) in a hybrid modeling approach resulted in a 2.1-fold increase in chlorophyll production. Optimized conditions included a NaNO3 concentration of 128.52 mg/l, a 1:2 (Algae:Bacteria) ratio, a 6-day cultivation period, and a pH of 5.4, yielding 10.92 ± 0.88 mg/l chlorophyll concentration., 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 Elsevier Ltd. All rights reserved.)

The co-pelletization of microalgae with filamentous fungi was a promising approach for microalgae harvest. However, the real conditions of microalgae growth limited the arbitrary optimization of co-pellets formation with filamentous fungi. Therefore, it is urgent to develop an approach to manipulate the co-pelletization through treatment of A. niger spores. In this study, Aspergillus niger and Chlorella vulgaris were used as the model species of filamentous fungi and microalgae to investigate co-pellets formation using A. niger spores after by different pH solutions treatment, swelling, snailase treatment. The importance of spore treatments on C. vulgaris harvest in sequence was claimed based on response surface methodology analysis. The pH solutions treatment, swelling, snailase treatment of A. niger spore contributed 21.0%, 10.5%, 40.7% of harvest ratio of C. vulgaris respectively, which guided the application of spore treatment into co-pelletization. Treatment of spore was showed as an efficient approach to manipulate co-pelletization for microalgae harvest in diverse microalgae condition. This results promoted the application of co-pelletization technology in microalgae harvest of various conditions., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Characteristics of an acid soil cultivated with Urochloa brizantha cv. Marandu were evaluated in relation to two types of fertilization: a conventional one, chemical based on nitrogen and potassium, and a biofertilizer, based on microalgae biomass. The results were compared among three treatments, control, conventional, and biological fertilization, with seven replications each. The study evaluated microalgae community, total carbon and nitrogen contents, mineral nitrogen, and enzymatic activity. Chlorella vulgaris showed the highest organism density, which can be explained by its rapid growth and high resistance. The highest species diversity was detected in the control 1,380,938 org cm -3 and biological 1,841,250 org cm -3 treatments, with the latter showing a higher density of cyanobacteria, especially Pseudanabaena limnetica with 394,554 org cm -3 . The soil treated with chemical fertilization showed higher nitrate (9.14 mg NKg -1 NO 3 - -N) and potassium (52.32 mg dm -3 ) contents. The highest levels of sulfur (21.73 mg dm -3 ) were detected in the biological treatment. The chemical treatment showed higher activity of the enzymes acid phosphatase, acetylglucosaminidase, and sulfatase, while α-glucosidase and leucine aminopeptidase stood out in the biological treatment. Soil properties were not significantly affected by the treatments. The use of microalgae biomass derived from wastewater treatment from milking parlors was evaluated and presented as a promising biofertilizer for agriculture, following the line of recovering nutrient-rich wastes. In this sense, although many challenges need to be overcome, the results suggest that microalgal-based fertilizers could lead to low-impact agriculture.-3 ) were detected in the biological treatment. The chemical treatment showed higher activity of the enzymes acid phosphatase, acetylglucosaminidase, and sulfatase, while α-glucosidase and leucine aminopeptidase stood out in the biological treatment. Soil properties were not significantly affected by the treatments. The use of microalgae biomass derived from wastewater treatment from milking parlors was evaluated and presented as a promising biofertilizer for agriculture, following the line of recovering nutrient-rich wastes. In this sense, although many challenges need to be overcome, the results suggest that microalgal-based fertilizers could lead to low-impact agriculture., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

The interest by biofilm-based microalgae technologies has increased lately due to productivity improvement, energy consumption reduction and easy harvesting. However, the effect of light, one key factor for system's operation, received less attention than for planktonic cultures. This work assessed the impact of Photon Flux Density (PFD) on Chlorella vulgaris biofilm dynamics (structure, physiology, activity). Microalgae biofilms were cultivated in a flow-cell system with PFD from 100 to 500 [Formula: see text]. In the first stage of biofilm development, uniform cell distribution was observed on the substratum exposed to 100 [Formula: see text] while cell clusters were formed under 500 [Formula: see text]. Though similar specific growth rate in exponential phase (ca. 0.3 [Formula: see text]) was obtained under all light intensities, biofilm cells at 500 [Formula: see text] seem to be ultimately photoinhibited (lower final cell density). Data confirm that Chlorella vulgaris showed a remarkable capability to cope with high light. This was marked for sessile cells at 300 [Formula: see text], which reduce very rapidly (in 2 days) their chlorophyll-a content, most probably to reduce photodamage, while maintaining a high final cell density. Besides cellular physiological adjustments, our data demonstrate that cellular spatial organization is light-dependent., (© 2024. The Author(s).)

This study proposed an innovative approach using lipid-cored particles (LCPs) aimed at addressing the efficiency, cost, and environmental impact challenges in microalgae harvesting. Cetyltrimethylammonium bromide (CTAB) and chitosan (CS) were used to modify LCPs and to optimize efficiency and investigate the mechanisms of harvesting with Chlorella vulgaris. Results showed that a maximum harvesting efficiency of 97.14 % was achieved using CS-LCPs. Zeta potential and microscopic images revealed the presence of embedded CS-LCPs within microalgal flocs. Fractal dimension data suggested looser aggregates of CS-LCPs and Chlorella vulgaris, corroborated by Excitation-emission matrices (EEM) analysis further confirmation the presence of bridging networks. Moreover, life cycle assessment of five harvesting methods pointed freshwater ecotoxicity potential (FEP) and terrestrial ecotoxicity potential (TEP) as major environmental impacts, mainly from flocculant use, carrier production, and electricity consumption. Notably, LCPs showed the lowest global warming potential (GWP) at 1.54 kg CO 2 eq, offering a viable, low-carbon, cost-effective harvesting alternative., 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 Elsevier Ltd. All rights reserved.)

Energy- and time-consuming concentration steps currently limit the industrial application of microalgae. Compared to state-of-the-art technologies, magnetic separation shows a high potential for efficient harvesting of microalgae. This study presents a novel approach to combine pH-induced calcium phosphate precipitation with cheap natural magnetite microparticles for magnetic separation of the freshwater microalgae Chlorella vulgaris. Harvesting efficiencies up to 98% were achieved at moderate pH and low particle and calcium phosphate concentrations in a model medium. However, cultivation-dependent high loads of algogenic organic matter can severely inhibit flocculation and particle/algae interactions, requiring higher salt concentrations or pH. Harvesting efficiencies above 90% were still attainable at moderate pH with increased calcium phosphate concentrations of 10mM. Acidification of the suspension to pH 5 allows for simple and reversible particle recycling. The presented process provides a promising path to universal and cost-effective harvesting, advancing the utilization of microalgae as a sustainable bioresource., 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 Elsevier Ltd. All rights reserved.)

The efficient management and treatment of pharmaceutical industry wastewater (PIWW) have become a serious environmental issue due to its high toxicity. To overcome this problem, the present study deals with the phycoremediation of PIWW using Chlorella vulgaris microalga isolated from the Ganga River at Haridwar, India. For this, response surface methodology (RSM) and artificial neural network (ANN) tools were used to identify the best reduction of total phosphorus (TP) and total Kjeldahl's nitrogen (TKN) based pollutants along with the lipid production efficiency of C. vulgaris. Three different concentrations of pharmaceutical wastewater (0, 50, and 100%), operating temperatures (20, 25, and 30 °C), and light intensity (2000, 3000, and 4000 lx) were used to design the phycoremediation experiments having 6:18 h of dark/light period and reactor functional volume of 15L. Findings revealed that C. vulgaris was good enough to remove maximum TP (90.35%), TKN (83.55%) along 20.88% of lipid yield at 25.62 °C temperature, 60.73% PIWW concentration, and 4000 lx of light intensity, respectively. Based on the model performance and validation results, ANN showed more accuracy as compared to the RSM tool. Therefore, the findings of this study showed that C. vulgaris is capable of treating PIWW efficiently along with significant production of lipid content which can further be used in various applications including biofuel production., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Research on renewable energy from microalgae has led to a growing interest in porous substrate photobioreactors, but their widespread adoption is currently limited to pure microalgal biofilm cultures. The behavior of microalgal-bacterial biofilms immobilized on microporous substrates remains as a research challenge, particularly in uncovering their mutualistic interactions in environment enriched with dissolved organic matter. Therefore, this study established a novel culture platform by introducing microalgal-derived bio-coating that preconditioned hydrophilic polyvinylidene fluoride membranes for the microalgal-bacterial biofilm growth of freshwater microalgae, Chlorella vulgaris ESP 31 and marine microalgae, Cylindrotheca fusiformis with bacteria, Escherichia coli. In the attached co-culture mode, the bio-coating we proposed demonstrated the ability to enhance microalgal growth for both studied species by a range of 2.5 % to 19 % starting from day 10 onwards. Additionally, when compared to co-culture on uncoated membranes, the bio-coating exhibited a significant bacterial growth promotion effect, increasing bacterial growth by at least 2.35 times for the C. vulgaris-E. coli co-culture after an initial adaptation phase. A significant increase of at least 72 % in intracellular biochemical compounds (including chlorophyll, polysaccharides, proteins, and lipids) was observed within just five days, primarily due to the high concentration of pre-coated organic matter, mainly sourced from the internal organic matter (IOM) of C. fusiformis. Higher accumulation of organic compounds in the bio-coating indirectly triggers a competition between microalgae and bacteria which potentially stimulate the production of additional intra-/extra-organic substances as a defensive response. In short, insight gained from this study may represent a paradigm shift in the ways that symbiotic interactions are promoted to increase the yield of specific bio-compounds with the presence of bio-coating., 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 Elsevier B.V. All rights reserved.)

Microalgae have been widely recognized as a promising feedstock for sustainable biofuels production to tackle global warming and pollution issues related to fossil fuels uses. This study identified and analyzed indigenous microalgae strains for biodiesel production, specifically Chlorella vulgaris and Coelastrella thermophila var. globulina, from two distinct locations in Algeria. Molecular identification confirmed their identity, and the microalgae exhibited notable growth characteristics. Local Chlorella vulgaris and Coelastrella thermophila var. globulina showed good growth and high biomass yield, compared to Chlorella vulgaris CCAP211/11B reaching a weight of 1.48 g L -1 , 1.95 g L -1 , and 2.10 g L -1 , respectively. Lipids content of local Chlorella vulgaris, Coelastrella thermophila var. globulina, and Chlorella vulgaris CCAP211/11B, were found to be 31.39 ± 3.3%, 17 ± 2.26%, and 19 ± 0.64%, respectively. Chlorella vulgaris stood out as a candidate for biodiesel production due to its equilibrium between SFA and PUFA (43.24% and 45.27%). FAs are predominated by SFA and MUFA for Coelastrella thermophila var. globulina with value of 81.49% (SFA+MUFA). Predicted biodiesel qualities comply with ASTM6751 and EN14214 standards. Studied microalgae have therefore a promising potential for biodiesel production. However, optimising cultivation conditions is necessary to enhance biomass and lipids yield at a large scale., (© 2023 Wiley-VCH GmbH.)

Microalgal biomass has been considered the third-generation biofuel production feedstock, but microalgae-derived biochar still needs to be thoroughly understood. This study aims to evaluate the production and physicochemical properties of microalgae-derived hydrochar produced by hydrothermal carbonization (HTC) process by comparison with pyrochar produced by dry thermal carbonization (DTC) process for environmental applications. Microalgal biochar was produced with commercially available Chlorella vulgaris microalgae using HTC and DTC processes under various temperature conditions. Pyrochar presented higher pH, ash contents, porosity, and surface area than hydrochar. Hydrochar gave more oxygen-containing functional groups on the surface and higher lead adsorption than pyrochar, making the microalgal hydrochar applicable in soil amendment and various environmental remediations. HTC could be an economically feasible thermochemical process for microalgal biochar production. It can produce hydrochar with high production yield from wet microalgae at low temperatures without a drying process., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

As pollution has attracted attention due to its wide distribution. An environmental concern that may be overlooked is that NPs additives are easily released into the environment due to their physical combination with NPs. However, the knowledge gaps still exist about the interfacial reactions of NPs and the additives (e.g. flame retardants) and the joint ecological effect. In the present study, fourier transform infrared (FTIR) spectrometer coupled with 2D correlation spectroscopy (2D-COS) analysis revealed the interfacial reactions between polystyrene nanoplastics (PSNPs) and Dechlorane Plus (DP). Results showed that carbon‑oxygen bonds and carbon‑chlorine bonds were the important binding sites during adhesion and DP could reduce the colloidal stability. Single and joint ecological effects of PSNPs and DP on the microalgae Chlorella vulgaris were further deliberated. Reduced photosynthetic efficiency (reduced F v /F m by 0.03 %), higher growth inhibition (16.15 %) and oxidative damage (increased ROS by 152 %) were observed in algae under co-exposure. Notably, DP could significantly increase the attachment of PSNPs to the surface of the algae. Metabolomics further revealed that co-exposure significantly down-regulated amino acid metabolism and tricarboxylic acid cycle (TCA) cycle, and up-regulated fatty acid metabolism. The present study provides new insights into the risk assessment of NPs in aquatic environment by investigating the interfacial reaction mechanism and combined ecotoxicity of NPs and additives., Competing Interests: Declaration of competing interest All authors declare that they have no known competing financial interests or personal relationships that could appear to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Bio-coating, a recent and promising approach in attached microalgal cultivation systems, has garnered attention due to its efficiency in enhancing immobilized algal growth, particularly in submerged cultivation systems. However, when the cells are cultured on thin solid microporous substrates that physically separate them from the nutrient medium, it remains unclear whether the applied bio-coatings still have a significant impact on algal growth or the subsequent rates of algal organic matter (AOM) release. Therefore, this current work investigated the role of bio-coatings on the microalgal monoculture growth of one freshwater species, Chlorella vulgaris ESP 31, and one marine species, Cylindrotheca fusiformis on a hydrophilic substrate, polyvinylidene fluoride membrane in a permeated cultivation system. Wide range of bio-coating sources were adapted, with the result demonstrating that bacteria-derived coating promoted algal growth by as high as 140% when compared with the control group for both species. Interestingly, two distinct adaptation mechanisms were observed between the species, with only C. fusiformis demonstrating a positive correlation between cell growth and AOM productivity, particularly in its extracellularly bound fractions. It is worth noting that despite this specific fraction exhibiting the lowest content among all; it displayed significant relevance in terms of AOM productivity. High extracellular protein-to-polysaccharide ratio (>5.7 fold) quantified on bacterial intracellular exudate-coated membranes indirectly revealed an underlying symbiotic microalgal-bacterial interaction. This is the first study showing how bio-coating influenced AOM yield without any physical interaction between microalgae and bacteria. It further confirms the practical benefits of bio-coating in attached cultivation systems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)