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

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.)

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.

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.)

In this study, the growth characteristics of microalgae cultured with different carbon sources were analyzed, and the flocculation characteristics under the influence of carbon sources were evaluated using three typical flocculants. The results showed that the organic carbon sources could significantly increase the content of extracellular proteins in microalgae. Specifically, the extracellular protein concentrations of microalgae cultured with pure BG-11, ethanol, sodium acetate and glucose were 18.2 29.2, 97.3, and 34.7 mg/g, respectively. During the flocculation process, microalgae cultured with sodium acetate exhibited a weak response to the flocculant because of excessive extracellular proteins inhibited flocculation. In addition, the flocculation efficiency was also less than 50.0% cultured with sodium acetate in all pH test ranges when alum and chitosan were used as flocculants. It could be inferred that the flocculant initially happened to charge neutralization with the negatively charged proteins in the solution and then bridged the charges with the microalgae. These findings provide insights into the effects of different carbon sources on microalgal flocculation, promising organic integration of microalgae wastewater treatment and harvesting., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The multiple microalgal collaborative treatment of domestic wastewater has been extensively investigated, but its whole life cycle tracking and consequent potential have not been fully explored. Herein, a dual microalgal system was employed for domestic wastewater treatment, tracking the variation in microalgal growth and pollutants removal from shake flask scale to 18 L photobioreactors scales. The results showed that Chlorella sp. HL and Scenedesmus sp. LX1 combination had superior growth and water purification performance, and the interspecies soluble algal products promoted their growth. Through microalgae mixing ratio and inoculum size optimized, the highest biomass yield (0.42 ± 0.03 g/L) and over 91 % N, P removal rates were achieved in 18 L photobioreactor. Harvested microalgae treated in different forms all promoted wheat growth and suppressed yellow leaf rate. This study provided data support for the whole process tracking of dual microalgal system in treating domestic wastewater and improving wheat 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 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.)

Heterotrophic cultivation of Chlorella has achieved commercial success, but the application of Chlorella biomass is still limited due to the high cost of biomass production. In this study, an effective and industrially scalable heterotrphic cultivation technology has been developed for a production strain Chlorella sorokiniana GT-1. Under the optimized culturing conditions, the ultrahigh biomass concentration of 271 and 247 g L -1 was achieved in 7.5 L bench-scale and 1000 L pilot-scale fermenters, respectively. Technoeconomic (TE) analysis indicated that the production cost of C. sorokiniana GT-1 could be reduced to $1601.27 per ton of biomass if the biomass concentration reached 200 g L -1 , which is 24.2% lower than that of the reported highest Chlorella biomass production through fermentation with the same TE model. Under the same growth conditions, the maximum biomass concentration of a low-starch mutant SLM2 was reduced to 93 g L -1 , which was 54% lower than that of the wild type, indicating the capabilities of C. sorokiniana GT-1 cells in accumulating large amounts of starch are essential for achieving the ultrahigh-cell-density under the heterotrophic conditions. In addition, the ultrahigh-cell-density growth potential of C. sorokiniana GT-1 cells was inferred to be related to the intrinsic biological characteristics including the tolerance to low dissolved oxygen and a moderate doubling time under the heterotrophic conditions as well. The breakthrough in cultivation technology is promising for Chlorella industry and would expand its applications in food and feed., (© 2021 Wiley Periodicals LLC.)

Ionic liquids are widely used for lipid and pigment extractions from microalgae. It is possible that ionic liquids are discharged into environments. The evaluation of growth performance and antioxidative response of ionic liquids to microalgae is helpful to explore the stress regulation mechanism and investigate possible environmental risk. Ionic liquids induce production of reactive oxygen species (ROS) to microalgae. These oxidative stresses are possible from cations, anions, and salinity. In this study, the growth inhibitions of [BMIM]Br, [BMIM]Cl, [EMIM]Cl, and [EMIM]EtOSO 3 to Anabaena cylindrica, Chlorella pyrenoidesa, and Dunaliella salina were evaluated. It was interesting that Br - and two kinds of cations, [BMIM] and [EMIM], had significant effects on growth inhibitions of these microalgae. IC50 values of these ionic liquids for A. cylindrica, C. pyrenoidesa, and D. salina were also estimated based on the results of growth inhibitions. It was proved that [EMIM]Cl is relatively harmless to C. pyrenoidesa and D. salina, and [EMIM]EtOSO 3 is relatively or practically harmless to C. pyrenoidesa. [BMIM]Br and [BMIM]Cl are practically harmless to A. cylindrica and C. pyrenoidesa, and relatively harmless to D. salina. More than 0.8 g/L [EMIM]EtOSO 3 led to bleaching of both A. cylindrica and D. salina at 48 h which was shown that the anion, EtOSO 3 - , had higher inhibition to A. cylindrica and D. salina than Cl - . In addition, high concentration of ionic liquids led to reductions of chlorophyll content in these three kinds of microalgae, increase of ROS levels and malondialdehyde contents for most of the cases. High concentration of ionic liquids also increased the activities of superoxide dismutase in three kinds of microalgae. There were positive correlations between ROS levels or MDA content, and inhibitions ratios of these ionic liquids to microalgae except [EMIM]Cl to A. cylindrica. These antioxidant enzymes were beneficial for reducing the ROS induced by ionic liquids.

The selection of a suitable growth regime can increase the physiological performance of microalgae and improve bioprocess based on these microorganisms from agro-industrial residues. Thus, this study assessed the biotechnology capacity-biomass production, biochemical composition, and nutrient uptake-from tequila vinasses (TVs) as the nutrient source of three indigenous microalgae-Chlorella sp., Scenedesmus sp., and Chlamydomonas sp.-cultured under heterotrophic and mixotrophic conditions. The results demonstrated that under the mixotrophic regime, the three microalgae evaluated reached the highest nitrogen uptake, biomass production, and cell compound accumulation. Under this condition, Chlorella sp. and Scenedesmus sp. showed the highest nutrient uptake and biomass production, 1.7 ± 0.3 and 1.9 ± 0.3 g L -1 , respectively; however, the biochemical composition, mainly carbohydrates and proteins, varied depending on the microalgal strain and its growth regime. Overall, our results demonstrated the biotechnological capacity of native microalgae from TVs, which may vary not only depending on the microalgal strain but also the culture strategy implemented and the characteristics of the residue used, highlighting-from a perspective of circular bio-economy-the feasibility of implementing microalgal bioprocess to reuse and valorize the nutrimental composition of TVs through biomass and high-valuable metabolite production, depicting a sustainable strategy for tequila agro-industry in Mexico.

The focus of this research is spectral shifting of light using phycocyanin solution extracted from the blue-green microalga Spirulina platensis in order to increase biomass productivity of the green microalga Chlorella sp. Also, lipid and chlorophyll content of the green alga was investigated. With regard to the shift of the spectrum with the phycocyanin solution, a double layer flat panel photobioreactor and two different spectral shifting strategies were used. In each strategy, the effect of two different concentrations of the solution was investigated. In the first strategy, the light passes through the chamber containing the solution and then enters the microalga culture chamber. In the second strategy, the light first enters the culture chamber and then enters the chamber containing phycocyanin pigment. The results showed that the use of phycocyanin pigment by both strategies increased the biomass productivity (P) and the specific growth rate (μ max ) with a significant difference compared to the control system; the increase in P for first strategy was up to about 69%. Moreover, the use of phycocyanin solution with a lower concentration had a greater effect on the increase of total chlorophyll content; however, the solution with a higher concentration was more successful in the production of cell lipid content. Using the phycocyanin solution as spectral converter in a double layer flat panel photobioreactor increased the biomass productivity and chlorophyll content.

Flat panels are the most spread type of photobioreactors for studying light effects on a microalgae culture. Their low thickness, usually between 1 and 3 cm, aims at ensuring light homogeneity across the culture. Yet because optical density has to remain very low, studies are still limited to low cell density cultures. To alleviate this problem, even thinner photobioreactors can be designed. Nevertheless, thin flat panel reactors are very prone to induce high shear stress. This work aimed at designing a new millimeter thin panel photobioreactor to study light effects on Chlorella and Scenedesmus genera. We proposed a numerical workflow that is capable of assessing the shear stress intensity in such a reactor. The numerical predictions were validated at three different levels: 2D preliminary simulations were able to reproduce bubble commonly known behaviors; close to the nozzle, the predictions were successfully confronted to shadowgraphy experimental reference; at the mini bioreactor scale, experimental and numerical mixing were found to be close. After these throughout validations, shear stress intensity in the photobioreactor was calculated over 1000 Lagrangian tracers. The experienced shear stress was agglomerated at the population level. From the computed shear stress, it was possible to choose the minimal reactor thickness that would not hinder cell growth., (© 2020 International Union of Biochemistry and Molecular Biology, Inc.)

As typical endocrine disrupters, nonylphenol (NP) and octylphenol (OP) are emerging pollutants that have attracted wide attention. This study investigated the toxicity effects of NP and OP on microalgae Chlorella pyrenoidosa and Scenedesmus obliquus, particularly on their growth inhibition, photosynthetic pigment, chlorophyll fluorescence, and superoxide dismutase and malondialdehyde levels. Results showed that the 96 h EC 50 of NP and OP was 2.89 and 5.21 mg/L on C. pyrenoidosa, respectively, and 1.54 and 8.48 mg/L on S. obliquus, respectively. NP exerted a stronger inhibitory effect on cell growth, photosynthesis, and PSII activity, and it contributed more oxidative stress on C. pyrenoidosa than on S. obliquus. By contrast, OP exerted a stronger inhibitory effect on S. obliquus than on C. pyrenoidosa. Furthermore, the toxicity of OP to the tested microalgae was lower than that of NP. Principal component analysis (PCA) and Pearson's correlation indicate that the accumulation of reactive oxygen species is the dominant mechanism of NP and OP cellular toxicity. The principal components of NP and OP affecting microalgae are distinct in the PCA plot, and different endocrine disrupters have varying chemical-specific influences on algal cells. This study confirmed that the toxicity of NP and OP to microalgae C. pyrenoidosa and S. obliquus is chemical- and species-specific. These findings should be considered when assessing the health risk of environmental pollution., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Chlorella sorokiniana CY-1 was cultivated using palm oil mill effluent (POME) in a novel-designed photobioreactor (NPBR) and glass-made vessel photobioreactor (PBR). The comparison was made on biomass and lipid productions, as well as its pollutants removal efficiencies. NPBR is transparent and is developed in thin flat panels with a high surface area per volume ratio. It is equipped with microbubbling and baffles retention, ensuring effective light and CO 2 utilization. The triangular shape of this reactor at the bottom serves to ease microalgae cell harvesting by sedimentation. Both biomass and lipid yields attained in NPBR were 2.3-2.9 folds higher than cultivated in PBR. The pollutants removal efficiencies achieved were 93.7% of chemical oxygen demand, 98.6% of total nitrogen and 96.0% of total phosphorus. Mathematical model revealed that effective light received and initial mass contributes toward successful microalgae cultivation. Overall, the results revealed the potential of NPBR integration in Chlorella sorokiniana CY-1 cultivation, with an aim to achieve greater feasibility in microalgal-based biofuel real application and for environmental sustainability.

Heterotrophic or mixotrophic culture of microalgae is feasible alternative approach to avoid light limitation in autotrophic culture. However, only a few kinds of organic carbon sources are available for algal culture. Disaccharides, such as sucrose, are difficult to be utilized by microalgae under both heterotrophic and mixotrophic conditions. In this study, a symbiotic yeast was accidentally found in a contaminated algal suspension. The symbiotic yeast was isolated and identified as Cryptococcus sp. This yeast was able to extracellularly hydrolyze sucrose and accumulated monosaccharides in the medium. It can enhance algal growth using sucrose as the carbon source at both heterotrophic and mixotrophic modes when mix-cultured with Chlorella pyrenoidosa. The highest algal cell density of 118.8 × 10 6 and 151.2 × 10 6 cells/mL was achieved with a final algal percentage of 83.5 and 93.2% at heterotrophic and mixotrophic culture, respectively. Furthermore, the protein and lipid content was significantly enhanced by mix-culture C. pyrenoidosa with Cryptococcus YZU-1. The fatty acid accumulated in this co-culture system was suitable for the production of biodiesel. This symbiotic yeast solved the problem that C. pyrenoidosa cannot heterotrophically or mixotrophically utilize sucrose. A high algae density was obtained and the protein and lipid accumulation were also significantly enhanced. This study provided a novel approach for production of protein or lipid-rich biomass using sucrose or sucrose-rich wastes as the carbon source.

In a previous study, the sequential optimization and regulation of environmental parameters using the PhotoBiobox were demonstrated with high-throughput screening tests. In this study, we estimated changes in the biovolume-based composition of a polyculture built in vitro and composed of three algal strains: Chlorella sp., Scenedesmus sp., and Parachlorella sp. We performed this work using the PhotoBiobox under different temperatures (10-36°C) and light intensities (50-700 μmol/m -2 /s -1 ) in air and in 5% CO 2 . In 5% CO 2 , Chlorella sp. exhibited better adaptation to high temperatures than in air conditions. Pearson's correlation analysis showed that the composition of Parachlorella sp. was highly related to temperature whereas Chlorella sp. and Scenedesmus sp. showed negative correlations in both air and 5% CO 2 . Furthermore, light intensity slightly affected the composition of Scenedesmus sp., whereas no significant effect was observed in other species. Based on these results, it is speculated that temperature is an important factor in influencing changes in algal polyculture community structure (PCS). These results further confirm that the PhotoBiobox is a convenient and available tool for performance of lab-scale experiments on PCS changes. The application of the PhotoBiobox in PCS studies will provide new insight into polyculture-based ecology.

Microalgae have evolved mechanisms to respond to changes in copper ion availability, which are very important for normal cellular function, to tolerate metal pollution of aquatic ecosystems, and for modulation of copper bioavailability and toxicity to other organisms. Knowledge and application of these mechanisms will benefit the use of microalgae in wastewater processing and biomass production, and the use of copper compounds in the suppression of harmful algal blooms. Here, using electron microscopy, synchrotron radiation-based Fourier transform infrared spectroscopy, electron paramagnetic resonance spectroscopy, and X-ray absorption fine structure spectroscopy, we show that the microalga Chlorella sorokiniana responds promptly to Cu2+ at high non-toxic concentration, by mucilage release, alterations in the architecture of the outer cell wall layer and lipid structures, and polyphosphate accumulation within mucilage matrix. The main route of copper detoxification is by Cu2+ coordination to polyphosphates in penta-coordinated geometry. The sequestrated Cu2+ was accessible and could be released by extracellular chelating agents. Finally, the reduction in Cu2+ to Cu1+ appears also to take place. These findings reveal the biochemical basis of the capacity of microalgae to adapt to high external copper concentrations and to serve as both, sinks and pools of environmental copper., (© 2020 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

The aim of the study was to identify the optimum cultivation conditions for the microalgal growth and lipid production of the oleaginous microalga Chlorella pyrenoidosa Chick (IPPAS C2). Moreover, an appropriate NO 3 - concentration in the cultivation medium for maximized lipid accumulation was determined. The experimental design involved a biphasic cultivation strategy with an initial biomass accumulating phase under optimized light (400 μmol/m 2 per s), temperature (25 °C), and elevated CO 2 concentration in the air mixture (3%), followed by a mid-elevated CO 2 concentration (0.5%) for lipid induction. The highest lipid yields of 172.47 ± 18.1 and 179.65 ± 25.4 mg/L per day were detected for NO 3 - concentrations of 100 and 150 mg/L. The optimization approach presented here led not only to the maximization of lipid yield but also to the development of a biphasic cultivation strategy easily applicable to the cultivation process without the necessity for algal cell harvesting between the first and second cultivation phases.

A number of dairies in southern Idaho employed stationary inclined screens to separate large solid particles out of liquid dairy manure. In this way, the total solid content of the liquid dairy manure can drop about 20%. Solids in dairy wastewater cause high turbidities, which could block the incident light, a key factor in the microalgae cultivation process using wastewaters as culture media. In this study, screened liquid dairy manure was used as the microalgae Chlorella vulgaris culture media. The aim was to optimize the dilution folds for the best growth of Chlorella vulgaris and nutrients' reduction with a special focus on phosphorus removal and recovery. Four folds of dilution, designated as 5*, 10*, 15*, 20*, were applied to the liquid dairy manure to alleviate hindrance of the high turbidity together with the high ammonium. Microalgal cultivation removed a significant amount of turbidity and major nutrients. For differently diluted liquid dairy manures, although the initial turbidities varied a lot, the final removal rates were not significantly different, falling in the range of 88.11-91.73%. Chemical oxygen demand (COD) in the 5-fold diluted liquid dairy manure dropped from 6700 to 1200 mg/L, corresponding to a removal rate of 79.81%. For the 10-fold, 15-fold, and 20-fold diluted manures, Chlorella removed around 67-69% of the initial CODs. Total Kjeldahl nitrogen (TKN) was removed at rates ranging from 70.84 to 73.99% from the four differently diluted liquid dairy manures without significant differences. NH 4 -N was removed most efficiently by 88.92% from the 20-fold diluted liquid dairy manure, and the least at 68.65% from the 5-fold diluted one. Although the original total phosphorus (TP) concentrations were distinctive for each group, the TP removal rates stayed in the range of 52.16 to 65.22%. Scanning electron microscopy (SEM) and energy-dispersive spectrometry (EDS) analysis of the precipitates harvested from the microalgal cultivation suggested possible phosphate precipitate forms. The chelation of Ca or Mg cations by dissolved organic matter (DOM) under alkaline conditions caused by microalgae cultivation could explain the unsatisfactory phosphorus removals observed in this study.

The use of microalgal biomass as feedstock for biofuels has been discussed for decades as it provides a sustainable approach to producing fuels for the future. Nonetheless, its feasibility has not been established yet and various aspects of biomass applications such as CO 2 biofixation should also be explored. Therefore, in this study, the CO 2 biofixation and lipid/carbohydrate production potential of Chlorella sp. ABC-001 were examined under various nitrogen concentrations. The highest biomass productivity and CO 2 biofixation rate of 0.422 g/l/d and 0.683 g/l/d, respectively, were achieved under a nitrogen-rich condition (15 mM nitrate). Carbohydrate content was generally proportional to initial nitrate concentration and showed the highest value of 41.5% with 15 mM. However, lipid content showed an inverse relationship with nitrogen supplementation and showed the highest value of 47.4% with 2.5 mM. In consideration as feedstock for biofuels (bioethanol, biodiesel, and biogas), the sum of carbohydrate and lipid contents were examined and the highest value of 79.6% was achieved under low nitrogen condition (2.5 mM). For lipid-based biofuel production, low nitrogen supplementation should be pursued. However, considering the lower feasibility of biodiesel, pursuing CO 2 biofixation and the production of carbohydrate-based fuels under nitrogenrich condition might be more rational. Thus, nitrogen status as a cultivation strategy must be optimized according to the objective, and this was confirmed with the promising alga Chlorella sp. ABC-001.

In the present work, the effect of α-Fe 2 O 3 -nanoparticles (IONPs) supplementation at varying doses (0, 10, 20 and, 30 mg L -1 ) at the intermittent stage (after 12th day of growth period) was studied on the growth and biogas production potential of Chlorella pyrenoidosa. Significant enhancements in microalgae growth were observed with all the tested IONPs doses, the highest (2.94 ± 0.01 g L -1 ) being at 20 mg L -1 . Consequently, the composition of the biomass was also improved. Based on the precedent determinations, theoretical chemical oxygen demand (COD th ) as well as theoretical and stoichiometric methane potential (TMP, and SMP) were also estimated. The COD th , TMP, SMP values indicated IONPs efficacy for improving biogas productivity. Further, the biochemical methane potential (BMP) test was done for IONPs supplemented biomass. The BMP test revealed up to a 25.14% rise in biogas yield (605 mL g -1 VS fed ) with 22.4% enhanced methane content for 30 mg L -1 IONPs supplemented biomass over control. Overall, at 30 mg L -1 IONPs supplementation, the cumulative enhancements in biomass, biogas, and methane content proffered a net rise of 98.63% in biomethane potential (≈ 2.86 × 10 4 m 3  ha -1 year -1 ) compared to control. These findings reveal the potential of IONPs in improving microalgal biogas production.

The effect of light wavelengths on the physiological, biochemical and lutein content of the microalgal consortia Chlorella variabilis and Scenedesmus obliquus was evaluated using different light sources. Among different light treatments, cool-white fluorescent light produced the highest biomass of 673 mg L -1 with a specific growth rate of 0.75 day -1 followed by blue (500 mg L -1 ; 0.73 day -1 ). The chlorophyll content was enhanced under blue light (10.7 mg L -1 ) followed by cool fluorescent light (9.3 mg L -1 ), whereas the lutein productivity was enhanced under cool fluorescent light (7.22 mg g -1 ). Protein content of the microalgal consortia was enhanced under all light treatments with the highest protein accumulation under cool-white fluorescent light (~56% of dry mass) closely followed by amber light (52% of dry mass), whereas the carbohydrate content was higher under amber light (~35% of dry mass). The results revealed that the consortia could grow well on diluted dairy wastewater thereby reducing the cost of algal production when compared with the use of inorganic media and a two-phase culture process utilizing cool fluorescent and amber light could be employed for maximizing algal biomass and nutrient composition with enhanced lutein production. The study also emphasizes on the economic efficiency of LED lights in terms of biomass produced based on the modest electricity consumed and the importance of using amber light for cultivating microalgae for its nutrient content which has seldom been studied.

In order to reduce the flow resistance of the thin-film flat plat photobioreactor (FPPBR) and make it more suitable for mass microalgae cultivation, the channel diameter was modified to 0.06 m and the thin-film FPPBR consisted of 10 parallel shunt-wound channels. A thin-film FPPBR system with 100 modified FPPBRs was constructed and used for Chlorella sp. cultivation in desertification areas (Ordos, China) from July to September of 2018. The pressure drop of the modified FPPBR system decreased significantly and the microalgae showed much higher productivity. The pressure drop was about 11.8 kPa when the liquid velocity was 0.238 m s -1 . The final biomass concentration and area productivity reached 2.01 g L -1 and 49.79 g m -2  day -1 respectively, and the yearly productivity of Chlorella sp. was estimated to be about 15.24 t ha -2 year -1 . The results demonstrated that high productivity of Chlorella sp. could be achieved in the improved FPPBR system in desertification areas and the improved FPPBR system was feasible for mass cultivation of microalgae in the commercial application., (Copyright © 2019 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Microalgae accumulate bioavailable selenium-containing amino acids (Se-AAs), and these are useful as a food supplement. While this accumulation has been studied in phototrophic algal cultures, little data exists for heterotrophic cultures. We have determined the Se-AAs content, selenium/sulfur (Se/S) substitution rates, and overall Se accumulation balance in photo- and heterotrophic Chlorella cultures. Laboratory trials revealed that heterotrophic cultures tolerate Se doses ∼8-fold higher compared to phototrophic cultures, resulting in a ∼2-3-fold higher Se-AAs content. In large-scale experiments, both cultivation regimes provided comparable Se-AAs content. Outdoor phototrophic cultures accumulated up to 400 μg g -1 of total Se-AAs and exhibited a high level of Se/S substitution (5-10%) with 30-60% organic/total Se embedded in the biomass. A slightly higher content of Se-AAs and ratio of Se/S substitution was obtained for a heterotrophic culture in pilot-scale fermentors. The data presented here shows that heterotrophic Chlorella cultures provide an alternative for Se-enriched biomass production and provides information on Se-AAs content and speciation in different cultivation regimes.

Microalgae have long been considered as potential biological feedstock for the production of wide array of bioproducts, such as biofuel feedstock because of their lipid accumulating capability. However, lipid productivity of microalgae is still far below commercial viability. Here, a glucose-6-phosphate dehydrogenase from the oleaginous microalga Nannochloropsis oceanica is identified and heterologously expressed in the green microalga Chlorella pyrenoidosa to characterize its function in the pentose phosphate pathway. It is found that the G6PD enzyme activity toward NADPH production is increased by 2.19-fold in engineered microalgal strains. Lipidomic analysis reveals up to 3.09-fold increase of neutral lipid content in the engineered strains, and lipid yield is gradually increased throughout the cultivation phase and saturated at the stationary phase. Moreover, cellular physiological characteristics including photosynthesis and growth rate are not impaired. Collectively, these results reveal the pivotal role of glucose-6-phosphate dehydrogenase from N. oceanica in NADPH supply, demonstrating that provision of reducing power is crucial for microalgal lipogenesis and can be a potential target for metabolic engineering., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Water-soluble proteins extracted from the heterotrophically cultivated microalga Chlorella protothecoides have been shown to have a good solubility over a broad pH range, which makes them a promising candidate for beverage formulations. This study investigated the sensory properties of dispersions of a protein-rich extract from C. protothecoides at neutral and pH 3., Results: Sensory acceptance tests of the pure extract revealed an overall low acceptance at pH 7 without sucrose addition. Sensory acceptance was significantly (P ≤ 0.05) increased by lowering the pH to 3 with citric acid, and the addition of 50 g kg -1 sucrose. Here, overall positive sensory acceptance ratings were achieved up to a protein extract concentration of 40 g kg -1 . Basic taste evaluations showed only low bitterness scores and no significant (P > 0.05) increase in bitterness with decreasing pH., Conclusion: It is suggested that protein-rich extracts from C. protothecoides have promising sensory properties in beverage formulations. © 2019 The Authors. Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2019 The Authors. Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Environmental factors such as temperature and light are the most determinants in the photosynthetic productivity in microalgae. However, under extreme of these conditions, certain resistant microalgae strains possess additional abilities such as growth in the presence of high concentrations of metals and some can improve in combinations of more than one abiotic stress. Therefore, the aim of this research was to evaluate the efficiency in photosynthetic production through the oxygen balance to variations in photon intensity, and under temperature changes in a Cd-resistant strain (Dc RCd100 ) compared to the wild-type strain (Dc1M wt ) of Dictyosphaerium chlorelloides. The results showed that the Dc RCd100 strain has the maximum efficiency at 200  μmol m -2 s -1 on photosynthesis net (Pn) (96.32 ± 3.63% nmol O 2 ml -1 min -1 ) as the threshold light saturation, and an adaptation to maintain this maximum photosynthetic gross (Pg) rate at 30 °C (94.99 ± 10.03% nmol O 2 ml -1 min -1 ) due to possible modifications in the photosynthetic apparatus that is reflected in the net evolution rate of O 2 to deal with such evaluated conditions. While, Dc1M wt strain its maximum photosynthetic efficiency was at 300 μmol m -2 s -1 and 21 °C (97.72 ± 2.99 and 99.85 ± 0.30%nmol O 2 ml -1 min -1 , respectively) and in optimal response to the oxygen balance that is normally achieved by this mesophilic genus. These results provide a new prediction of mechanisms in the oxygen evolution in photosynthesis that rules the correlation between resistance and adaptation to extreme abiotic conditions in metal resistant strains of eukaryotic microalgae., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

This study investigated the nutrient content and reuse potential of wastewater generated during hydrothermal liquefaction of microalgal biomass. The hydrothermal liquefaction reaction was tested at 270, 300, 330, and 345 °C to determine the effect of temperature on the formation of non-biodegradable dissolved organic nitrogen (nbDON). Total nitrogen, ammonium, color, and toxicity were selected as key characteristics for the reuse of hydrothermal liquefaction wastewater. Results indicated that a higher concentration of nbDON 5 (nbDON defined with a 5 day growth assay) and more diverse heterocyclic N-containing organic compounds were associated with greater toxicity as measured by a growth rate assay. For the tested temperature ranges, the total nitrogen content of the hydrothermal liquefaction wastewater slightly decreased from 5020 ± 690 mg L -1 to 4160 ± 120 mg L -1 , but the % nbDON 5 fraction increased from 57 ± 3 %DON to 96 ± 5 %DON. The temperature of hydrothermal liquefaction reactions can be optimized to maximize carbon conversion and nitrogen recovery., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The heterotrophic cultivation of microalgae has a number of notable advantages, which include allowing high culture density levels as well as enabling the production of biomass in consistent and predictable quantities. In this study, the full potential of Chlorella sp. HS2 is explored through optimization of the parameters for its heterotrophic cultivation. First, carbon and nitrogen sources were screened in PhotobioBox. Initial screening using the Plackett-Burman design (PBD) was then adopted and the concentrations of the major nutrients (glucose, sodium nitrate, and dipotassium phosphate) were optimized via response surface methodology (RSM) with a central composite design (CCD). Upon validation of the model via flask-scale cultivation, the optimized BG11 medium was found to result in a three-fold improvement in biomass amounts, from 5.85 to 18.13 g/L, in comparison to a non-optimized BG11 medium containing 72 g/L glucose. Scaling up the cultivation to a 5-L fermenter resulted in a greatly improved biomass concentration of 35.3 g/L owing to more efficient oxygenation of the culture. In addition, phosphorus feeding fermentation was employed in an effort to address early depletion of phosphate, and a maximum biomass concentration of 42.95 g/L was achieved, with biomass productivity of 5.37 g/L/D.

Positive interactions such as facilitation play an important role during the biological colonization and species succession in harsh or changing environments. However, the direct evidence of such ecological interaction in microbial communities remains rare. Using common freshwater microalgae isolated from a High Rate Algal Pond HRAP treating wastewaters, we investigated with both experimental and modeling approaches the direct facilitation between two algal strains during the colonization phase. Our results demonstrate that the first colonization by microalgae under a severe chemical condition arose from the rapid growth of pioneer species such as Chlorella sorokiniana, which facilitated the subsequent colonization of low growth specialists such as Scenedesmus pectinatus. The pioneer species rapidly depleted the total available ammonia nitrogen favoring the specialist species initially inhibited by free ammonia toxicity. This latter species ultimately dominated the algal community through competitive exclusion under low nutrient conditions. We show that microbial successions are not only regulated by climatic conditions but also by interactions between species based on the ability to modify their growth conditions. We suggest that facilitation within the aquatic microbial communities is a widespread ecological interaction under a vast range of environmental stress.

Biogas, a gaseous effluent from the anaerobic digestion of organic waste, is considered an important source of energy, since it has a composition mainly of methane (CH 4 ; 55-75%) and CO 2 (20-60%). Today, CO 2 from biogas is an excellent carbon source to induce high microalgal biomass production; however, each microalga strain can have different optimal CO 2 concentrations for maximizing their bio-refinery capacity as well as different ability to endure stressful conditions of industrial effluents. This study assessed the bio-refinery capacity of Chlorella sp. and Scenedesmus sp., native of Lago de Chapala, Mexico, from biogas, as well as the effect of high CO 2 and methane concentrations on the physiological performance to grow, capture CO 2 and biochemical composition of both microalgae cultured under different biogas compositions. The results show that both microalgae have the biotechnological potential to endure biogas compositions of 25% CO 2 -75% CH 4 . Under this condition, the biomass production attained by Chlorella sp. and Scenedesmus sp. was 1.77 ± 0.32 and 2.25 ± 0.20 g L -1 , respectively, with a biochemical composition mainly of carbohydrates and proteins. Overall, this study demonstrates that both microalgae have the ability to endure the stressful biogas composition without affecting their physiological capacity to capture CO 2 and biosynthesize high-value metabolites. Moreover, it is worth highlighting the importance of screening wild-type microalgae from local ecosystems to determine their physiological capacity for each biotechnological application.

Microbial biomass which mostly generated from the microbial processes of bacteria, yeasts, and microalgae is an important resource. Recent concerns in microbial biomass production field, especially microbial lipid production for biofuel, have been focused towards the mixed culture of microalgae and yeast. To more comprehensive understanding of the mixed culture for microbial biomass, mono Chlorella pyrenoidosa, mono Yarrowia lipolytica and the mixed culture were investigated in the present work. Results showed that the mixed culture achieved significantly faster cell propagation of microalga and yeast, smaller individual cell size of yeast and higher relative chlorophyll content of microalga. The mixed culture facilitated the assimilation of carbon and nitrogen and drove the carbon flow to carbohydrate. Besides higher lipid yield (0.77 g/L), higher yields of carbohydrates (1.82 g/L), protein (1.99 g/L) and heating value (114.64 kJ/L) indicated the microbial biomass harvested from the mixed culture have more potential utilization in renewable energy, feedstuff, and chemical industry.

Background: Microalgae have been proposed as potential platform to produce lipid-derived products, such as biofuels. Knowledge on the intracellular carbon flow distribution may identify key metabolic processes during lipid synthesis thus refining culture/genetic strategies to maximize cell lipid productivity. A kinetic metabolic model simulating cell metabolic behavior and lipid production was first applied in the microalgae platform Chlorella protothecoides under heterotrophic condition. It combines both physiology and flux information in a kinetic approach. Cell nutrition, growth, lipid production and almost 30 metabolic intermediates covering central carbon metabolism were included and simulated., Results: Model simulations were shown to adequately agree with experimental data, which is suggesting that the proposed model copes with Chlorella protothecoides cells' biology. The dynamic metabolic flux analysis using the model showed a reversible starch flux from accumulation to decomposing when glucose reached depletion, while net lipid flux shows a quasi-constant rate. The sensitive flux parameters on starch and lipid metabolism suggested that starch synthesis is the major competing pathway that affects lipid accumulation in C. protothecoides. Flux analysis also demonstrated that high lipid yield under heterotrophic condition is accompanied with high lipid flux and low TCA activity. Meanwhile, the dynamic flux distribution also suggests a relatively constant ratio of glucose distributed to biomass, lipid, starch, nucleotides as well as pentose phosphate pathway., Conclusion: The model described not only experimental data, but also unraveled intracellular carbon flow distribution and identify key metabolic processes during lipid synthesis. Most of the metabolic kinetics also showed statistical significance for metabolic mechanism. Therefore, this study unravels the mechanisms of the glucose impact on the dynamic carbon flux distribution, thus improving our understanding of the links between carbon fluxes and lipid metabolism in C. protothecoides.

17β-estradiol (E2) is a steroid estrogen able to affect the reproduction of aquatic organisms even at extremely low concentrations. The behavior of E2 in the presence of chlorella algae was investigated in laboratory experiments. The results showed that the algae's growth was inhibited by 26% after 7 days of culturing in a 2.0 mg L -1 solution of E2. The 96 h EC 50 value of 21.46 mg L -1 reflected moderate toxicity. Even low concentrations of E2 were found to affect total chlorophyll and carotenoid levels after 7 and 10 days and to alter stress-generated enzymatic activity in the algae. The efficiency of chlorella's E2 degradation decreased with the increasing of E2 concentration, but 92% of the E2 can be removed from a 0.5 mg L -1 solution over 10 days. The degradation mechanism was speculated. The microalgae suffered relatively less growth inhibition at low E2 concentrations, and their removal effectiveness was then better. The data help to elucidate the interaction between chlorella algae and E2 in an aquatic environment., (Copyright © 2019. Published by Elsevier Inc.)

The successful production of microalgal biomass requires the precise coordination of many different steps. Cell harvesting is a central process in all methods currently used for the production of microalgal biomass. Therefore, improving the harvesting process itself, and using a harvesting method that is compatible with adjacent steps, is necessary to prevent problems that may occur during downstream processing. This study examined the potential of the cationic biopolymer ε-poly-L-lysine (ε-PLL) for use in the harvest of microalgae (Chlorella ellipsoidea). The effects of ε-PLL concentration and mixing intensity on flocculation efficiency and operating costs were determined. We found that ε-PLL was not toxic to microalgal cells at concentrations of up to 25 mg/L, based on the photosystem II quantum yield. A recovery rate of 95% was achieved using 19 mg/L ε-PLL, and the estimated harvest cost was 20 US$/ton of harvested biomass. Moreover, ε-PLL displayed antimicrobial properties, leaving the harvested biomass intact and pure. Therefore, the use of ε-PLL-induced flocculation appears to be an attractive option when harvesting microalgal biomass for use as low- and high-value commodities for humans or animals.

A mild and low-energy cell disruption method with high efficiency has growing application potential in both the extraction of high-value microalgal products and the inactivation of microalgal cells. Conventional technologies available have disadvantages including high energy consumption, the use of chemicals and so on. Here, this study developed an efficient microalgal cell disruption method using the copper oxide nanowire (CuONW)-modified three-dimensional (3D) copper foam electrodes with a low applied voltage. Electrodes with nanowires synthesized at 400 °C, the optimal preparation temperature, achieved efficient microalgal cell electroporation. Microalgal cells were completely inactivated and disrupted at the voltage of 2 V with the hydraulic retention time (HRT) of 10 s. Scanning electron microscopy (SEM) images showed obvious electroporation damage on the cell surface upon electroporation-treatment (2 V, 30 s). The amount of released cellular inclusion increased significantly with prolonged HRT and the energy consumption of this technology was only 0.014 kWh/kg via the treatment of 2 V and 10 s. This study provided a novel, energy-efficient and chemical-free technique for both microalgal products extraction and cell inactivation., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The present study reports the use of Citrus limetta (CL) residue for cultivating Chlorella sp. mixotrophically to augment production of biodiesel. The cultivation of Chlorella sp. using CL as media was carried out by employing a fed-batch technique in open tray (open tray+CL) and in software (BioXpert V2)-attached automated photobioreactor (PBR+CL) systems. Data showed the limit of nitrogen substituent and satisfactory organic source of carbon (OSC) in CL, causing > 2-fold higher lipid content in cells, cultivated in both the systems than in control. For the cells grown in both the systems, ≥ 3-fold enhancement in lipid productivity was observed than in control. The total fatty acid methyl ester (FAME) concentrations from lipids extracted from cells grew in PBR+CL and in open tray+CL techniques were calculated as 50.59% and 38.31%, respectively. The PBR+CL system showed improved outcomes for lipid content, lipid and biomass productivity, FAME characteristics and physical property parameters of biodiesel than those obtained from the open tray+CL system. The physical property parameters of biodiesel produced from algal cells grown in PBR+CL were comparable to existing fuel standards. The results have shown lower cold filter plugging point (- 6.57 °C), higher cetane number (58.04) and average oxidative stability (3.60 h). Collectively, this investigation unveils the novel deployment of CL as a cost-effective feedstock for commercialisation of biodiesel production.

CO 2 , SO 2 , and NO are the main components of flue gas and can cause serious environmental issues. Utilization of these compounds in oleaginous microalgae cultivation not only could reduce air pollution but could also produce feedstock for biodiesel production. However, the continuous input of SO 2 and NO inhibits microalgal growth. In this study, the toxicity of simulated flue gas (15% CO 2 , 0.03% SO 2 , and 0.03% NO, balanced with N 2 ) was reduced through automatic pH feedback control. Integrated lipid production and CO 2 fixation with the removal of SO 2 and NO was achieved. Using this technique, a lipid content of 38.0% DW was achieved in Chlorella pyrenoidosa XQ-20044. The lipid composition and fatty acid profile indicated that lipid production by C. pyrenoidosa XQ-20044 cultured with flue gas is suitable as a biodiesel feedstock; 81.2% of the total lipids were neutral lipids and 99.5% of the total fatty acids were C16 and C18. The ratio of saturated fatty acids to monounsaturated fatty acids in the microalgal lipid content was 74.5%. In addition, CO 2 , SO 2 , and NO from the simulated flue gas were fixed and converted to biomass and lipids with a removal efficiency of 95.9%, 100%, and 84.2%, respectively. Furthermore, the utilization efficiencies of CO 2 , SO 2 , and NO were equal to or very close to their removal efficiencies. These results provide a novel strategy for combining biodiesel production with biofixation of flue gas.

Microalgae harvesting via pH induced flocculation along with utilization of recovered medium after flocculation is one of the most economical methods for separating the microalgal biomass in order to reduce the dewatering cost. In this study, optimization of marine and freshwater microalgae flocculation by pH adjustment was investigated via central composite design methodology. One molar of KOH and NaOH solutions were used to increase the pH level of the microalgal culture. Increasing pH value of the medium provided the highest flocculation efficiency up to 92.63 and 86.18% with pH adjusted to 10.5 with KOH and NaOH solutions for marine microalgae Nannochloropsis oculata and freshwater microalgae Chlorella minutissima, respectively. Also, it was revealed that microalgae cells were still alive after flocculation process and their biochemical composition was not changed, and flocculated medium can be used again for the next microalgal production. According to the results, it can be said that this method is cheap and effective, simple to operate and provides the utilization of flocculated medium again., (© 2019 American Institute of Chemical Engineers.)

Benzotriazoles are among the most commonly found organic micropollutants in the aquatic environment. In this study, toxicity experiments were conducted in order to investigate the effects of different benzotriazoles on Chlorella sorokiniana growth. Four compounds were tested; 1H-benzotriazole (BTR), xylytriazole (XTR), 5-methyl-1H-benzotriazole (5TTR) and 5-chlorobenzotriazole (CBTR). The fate of these micropollutants was also studied under batch conditions and the effect of different mechanisms on their elimination was investigated. According to the results, the EC 50 values in single-substance toxicity experiments were calculated to 8.3 mg L -1 for BTR, 22 mg L -1 for 5TTR and 38.7 mg L -1 for CBTR. A slight inhibition on microalgae growth was noted at the maximum tested concentration of XTR (77 mg L -1 ), while no inhibition was observed when a mixture of target BTRs was tested at 200 μg L -1 . Calculation of the Risk Quotient (RQ) showed no possible ecological threat in the presence of 5TTR, XTR and CBTR, while RQ values close or higher than 1 were estimated for BTR. All target contaminants were significantly eliminated in microalgae experiments that lasted 16 days. Their removal efficiency ranged between 42.2 ± 3.1% (XTR) to 97.2 ± 0.9% (XTR), while their half-life values were estimated to 2.4 ± 0.5 days for 5TTR, 6.5 ± 0.6 days for BTR, 15.2 ± 1.4 days for CBTR and 20.7 ± 2.0 days for XTR. Photodegradation was the main mechanism affecting BTR, XTR and CBTR removal, while bioremoval processes enhanced 5TTR elimination. The addition of sodium acetate decreased the removal efficiency of BTRs possibly due to catabolite repression. This is the first study investigating the toxicity and fate of BTRs in microalgae cultures., (Copyright © 2019. Published by Elsevier B.V.)

A CO 2 microbubbles dissolver (CMD) was developed to facilitate dissolving inorganic carbon and strengthening mass transfer in a horizontal tubular photo-bioreactor system (HTPBRS), which enhanced microalgae biomass productivity with flue gas containing 15% CO 2 . The influence of pump power on the bubble formation and mixing effect was found to be more obvious than that of gas flow rate. Ceramic shell aerator was more favorable for reducing bubble diameter and enhancing mass transfer than traditional rubber strip aerator. Bubble formation time decreased by 53.4% and mixing time decreased by 68.9% in response to the increased pump power. When the base area ratio of ceramic shell aerator to dissolver in the HTPBRS increased, bubble formation time decreased by 19.6% and mass transfer coefficient increased by 80.9%. The biomass yield of microalgae Chlorella PY-ZU1 with ceramic shell aerator was 30% higher than that with rubber strip aerator in the HTPBRS., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The isolated microalga Chlorella sorokiniana BENHA721_ABO4 was grown in Bold's basal medium (BBM) as a control, municipal wastewater (WW), and wastewater enriched with BBM elements (WW+). Cultivation in WW+ showed the highest cell number which represented 25.3 and 47.3% over that grown in WW and BBM, respectively. However, rapid growth in WW+ was accompanied by significant reduction in lipid content. Due to lipid accumulation in WW, it showed the maximum significant lipid productivity of 16.2 mg L -1  day -1 . Microalgae cultivation in WW for 10 days showed 74.2, 83.3, and 78.0% removal efficiency for NO 3 -N, NH 3 -N and TP, respectively. In addition, growth in WW significantly reduced polyunsaturated fatty acids by 36.0% with respect to BBM in favor of monounsaturated fatty acids. The present results confirmed that C. sorokiniana isolate BENHA721_ABO4 grown in secondary effluent municipal wastewater offers real potential for future application in wastewater treatment and biodiesel production.

In this study, effects of different abiotic factors were studied on biomass and lipid yield of green microalga Chlorella minutissima (C. minutissima) Various concentrations of abiotic factors like nitrogen, phosphorus, glucose, iron, zinc, different values of pH, temperature, light intensity and different photoperiods were observed on the biomass growth and lipid yield of C. minutissima cultivated with modified CHU-13 medium. Initially, three cultivation media namely, Bold's basal medium (BBM), modified CHU-13 and blue-green-11 (BG-11) were used to culture C. minutissima in batch mode. Microalga cultivated with modified CHU-13 medium resulted in maximum biomass and lipid yield of 970 ± 0.21 and 356.63 ± 0.51 mg/L, respectively. To maximize biomass and lipid yield of microalga further, it was cultivated with modified CHU-13 medium and variation of above mentioned abiotic factors was done. Different biomass and lipid yields were achieved for different abiotic factors varied. Highest biomass of 1840.49 ± 0.62 mg/L was achieved with 12 g of glucose containing medium and highest lipid yield of 579.86 ± 0.76 mg/L was achieved with 0.3 g of nitrogen containing medium. GC-MS analysis of biodiesel obtained from C. minutissima biomass cultivated with modified CHU-13 medium shown the presence of C14:0, C16:0, C16:1, C18:0, C18:1, C18:2, C18:3, C20:0, C20:1 and C22:0. Properties of biodiesel obtained from C. minutissima were found in compliance with ASTM-6751-02 and European biodiesel standards EN14214. These results suggest that C. minutissima can be used as a potential biodiesel feedstock for microalgal biodiesel production.

Phenanthrene is one of the most abundant polycyclic aromatic hydrocarbons (PAHs) found in continental shelf environment of China and is on the EPA's Priority Pollutant list. In this study, the effects of phenanthrene on marine algal growth rate were determined after 96-h exposure at pH 6.0, 7.0, 8.0, 9.0, and 10.0 in seawater of salinity 35. Two measuring techniques to assess growth inhibition were also compared using prompt fluorescence and microscopic cell count. The results showed that the toxicity of phenanthrene increased significantly (p < 0.05) with decreasing pH, with the nominal concentration required to inhibit growth rate by 50%, EC 50 , decreasing from 1.893 to 0.237 mg L -1 as pH decreased from 9.0 to 6.0, with a decrease higher than 55% from 10.0 to 9.0. In addition, the nominal EC 50 values calculated in this study were at the same range of some environmental concentrations of phenanthrene close to areas of crude oil exploration. Based on the two measuring techniques, the results showed that cell count and fluorescence measurement were significantly different (p < 0.05), and the nominal EC 50 values calculated with cell count measurement were significantly higher than fluorescence measurement at pH 8.0, 9.0 and 10.0. In conclusion, the present studies confirmed that acidification of seawater could affect the toxicity of phenanthrene to this species of microalgae, and which encouraged further studies involving responses of marine organisms to ocean acidification.

The high cost of large-scale cultivation of microalgae has limited their industrial application. This study investigated the potential use of mixed biogas slurry and municipal wastewater to cultivate microalgae. Pig biogas slurry as the sole nutrient supplement, was assessed for the cultivation of Chlorella zofingiensis in municipal wastewater. Batch culture of various ratios of pig biogas slurry and municipal wastewater were compared. The characteristics of algal growth and lipid production were analyzed, and the removal rates of nitrogen and phosphate were examined. Results indicate that 8% pig bio-gas slurry in municipal wastewater, had a significant effect on microalgal growth. C. zofingiensis, with 2.5 g L -1 biomass, 93% total nitrogen and 90% total phosphorus removal. Lipid content was improved by 8% compared to BG11 medium. These findings show that mixing pig biogas slurry and municipal wastewater, without additional nutrition sources, allows efficient cultivation of C. zofingiensis. This is of high research and industrial significance, allowing cultivation of C. zofingiensis in mixed waste culture solution without additional nutrition sources., (Copyright © 2018. Published by Elsevier B.V.)

Microalgae are the most promising sources of protein, which have high potential due to their high-value protein content. Conventional methods of protein harnessing required multiple steps, and they are generally complex, time consuming, and expensive. Currently, the study of integration methods for microalgae cell disruption and protein recovery process as a single-step process is attracting considerable interest. This study aims to investigate the novel approach of integration method of electrolysis and liquid biphasic flotation for protein extraction from wet biomass of Chlorella sorokiniana CY-1 and obtaining the optimal operating conditions for the protein extraction. The optimized conditions were found at 60% (v/v) of 1-propanol as top phase, 250 g/L of dipotassium hydrogen phosphate as bottom phase, crude microalgae loading of 0.1 g, air flowrate of 150 cc/min, flotation time of 10 min, voltage of 20 V and electrode's tip touching the top phase of LBEF. The protein recovery and separation efficiency after optimization were 23.4106 ± 1.2514% and 173.0870 ± 4.4752%, respectively. Comparison for LBEF with and without the aid of electric supply was also conducted, and it was found that with the aid of electrolysis, the protein recovery and separation efficiency increased compared to the LBEF without electrolysis. This novel approach minimizes the steps for overall protein recovery from microalgae, time consumption, and cost of operation, which is beneficial in bioprocessing industry.

Mixotrophy is a metabolic strategy in which an organism is autotrophic and heterotrophic simultaneously. Considering that the aquatic environment provides several organic sources of carbon, it is probably common for microalgae to perform mixotrophy and not only photoautotrophy, but little is known about microalgae mixotrophy. The present work aimed at investigating the growth, photosynthetic activity, morphology, and biochemical composition of the microalga Chlorella sorokiniana in mixotrophic and photo-mixotrophic conditions, comparing it with photoautotrophy. The results showed pH changes after glucose addition, reaching pH 11.62 in mixotrophic and 10.47 in sequential photo-mixotrophic cultures, which limited the microalgal growth. Highest biomass was obtained in the mixotrophic culture in comparison with the sequential photo-mixotrophic one. Rapid light saturation curves showed that α (photosynthetic efficiency, 1.69) and relative electron transport rate (rETR; 565.61) were higher in the mixotrophic cultures, whereas the highest I k (irradiance saturation, 386.68) was obtained in the photoautotrophic ones. In the sequential photo-mixotrophic cultures, photosynthetic activity varied during glucose consumption, decreasing the maximum quantum yield F v /F m after glucose addition, indicating change in metabolism, from photoautotrophy to mixotrophy by the microalga. The results showed that the mixotrophic cultures had higher production of chlorophyll a (6.26 mg mL -1 ), cell density (6.62 × 10 7 cell mL -1 ), and lipids (0.06 pg μm -3 ). Sequential photo-mixotrophic cultures showed the highest biovolume (360.5 μm 3  cell -1 ) and total carbohydrates (0.026 pg μm -3 ). The protein concentration was 3.2 and 2.4 times higher in photoautotrophy and photo-mixotrophic growth, respectively, than in mixotrophy, but lipids were three times higher under mixotrophy. The biochemical changes we observed indicate that the microalga's plasticity in face of new environmental characteristics, such as the presence of organic carbon, can change the flow of energy through natural ecosystems.

This study sought to evaluate influence of nitrogen availability on cell growth, biomass composition, production, and the properties of polyhydroxyalkanoates during cultivation of microalgae Chlorella minutissima, Synechococcus subsalsus, and Spirulina sp. LEB-18. The cellular growth of microalgae reduced with the use of limited nitrogen medium, demonstrating that nitrogen deficiency interferes with the metabolism of microorganisms and the production of biomass. The biochemical composition of microalgae was also altered, which was most notable in the degradation of proteins and chlorophylls and the accumulation of carbonaceous storage molecules such as lipids and polyhydroxyalkanoates. Chlorella minutissima did not produce these polymers even in a nitrogen deficient environment. The largest accumulations of the polyhydroxyalkanoates occurred after a 15 days culture, with a concentration of 16% (dry cell weight) produced by the Synechococcus subsalsus strain and 12% by Spirulina sp. LEB-18. Polyhydroxyalkanoates produced by Synechococcus subsalsus and Spirulina sp. LEB-18 presented different thermal and physical properties, indicating the influence of producing strain on polyhydroxyalkanoates properties. The polymers obtained consisted of long chain monomers with 14 to 18 carbon atoms. This composition is novel, as it has not previously been found in PHAs obtained from Synechococcus subsalsus and Spirulina sp. LEB-18., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Actual biomass of microalgae was tested as a fermentation substrate for microbial production of 2-pyrone 4,6-dicarboxylic acid (PDC). Acid-hydrolyzed green microalgae Chlorella emersonii (algae hydrolysate) was diluted to adjust the glucose concentration to 2 g/L and supplemented with the nutrients of Luria-Bertani (LB) medium (tryptone 10 g/L and yeast extract 5 g/L). When the algae hydrolysate was used as a fermentation source for recombinant Escherichia coli producing PDC, 0.43 g/L PDC was produced with a yield of 20.1% (mol PDC/mol glucose), whereas 0.19 g/L PDC was produced with a yield of 8.6% when LB medium supplemented with glucose was used. To evaluate the potential of algae hydrolysate alone as a fermentation medium for E. coli growth and PDC production, the nutrients of LB medium were reduced from the algae hydrolysate medium. Interestingly, 0.17 g/L PDC was produced even without additional nutrient, which was comparable to the case using pure glucose medium with nutrients of LB medium. When using a high concentration of hydrolysate without additional nutrients, 1.22 g/L PDC was produced after a 24-h cultivation with the yield of 16.1%. Overall, C. emersonii has high potential as cost-effective fermentation substrate for the microbial production of PDC., (Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Green microalgae colonizing stone surfaces represent a major problem for the conservation of heritage monuments, since they lead to biodegradation and aesthetic issues. Previous studies in La Glacière show cave (France) have demonstrated that UV-C may have a strong effect on microalgae, thus leading to chlorophyll bleaching, which was increased when biofilms were maintained under VIS-light condition unlike to those maintained in the dark. To understand the physiological mechanisms underlying this response and in order to optimize in situ treatment, 30 kJ m -2 UV-C exposure times were applied to Chlorophyta Chlorella sp. and chlorophyll degradation kinetics were then monitored. UV-C irradiation was enough to inhibit photosynthesis and to directly kill all algal cells. Results also showed that chlorophyll a was degraded faster than chlorophyll b and that 14 h were necessary for complete degradation of all the present chlorophyll. In addition, our results highlighted the importance of visible light exposition after UV-C treatment which leading to chlorophyll bleaching. Irradiated algae cultivated in the dark were still green 5 days after treatment while cultivated samples in the light lost their green color after 14 h. An efficient UV-C treatment applicable to show caves and other heritage monuments was proposed.