Cultivation of the macrophyte Lemna minor and the microalgae Chlorella sorokiniana in thermal mineral waters: Biomass characteristics, radioisotopes and heavy metals content.

Microalgae and macrophytes are commonly used as human and animal food supplements. We examined the cultivation of the microalgae Chlorella sorokiniana and the duckweed Lemna minor in thermal waters under batch and sequencing batch conditions and we characterized the produced biomass for the presence of essential nutrients as well as for heavy metals and radioisotope content. The highest specific growth rate for the microalgae was observed when 5 or 15 mg/L N were supplemented while the optimal conditions for Lemna minor were observed in the co-presence of 5 mg/L N and 1.7 mg/L P. Lemna minor presented higher concentrations of proteins and lipids comparing to the studied microalgae. Both organisms contained high amounts of lutein (up to 1378 mg/kg for Lemna minor) and chlorophyll (up to 1518 mg/kg for Lemna minor) while ß-carotene and tocopherols were found at lower concentrations, not exceeding a few tens of mg/kg. The heavy metal content varied between the two species. Lemna minor accumulated more Cd, Cu, K, Mn, Na, Ni, and Zn whereas Al, Ca and Mg were higher in Chlorella sorokiniana. Both organisms could be a significant source of essential metals but the occasional exceedance of the statutory levels of toxic metals in food products raises concern for potential risk to either humans or animals. Application of gamma-spectroscopy to quantify the effective dose to humans from 228Ra, 226Ra and 40K showed that Chlorella sorokiniana was well under the radiological limits while the collected mass of Lemna minor was too small for radiological measurements with confidence.

Bioremediation of hemotoxic and oxidative stress induced by polyethylene microplastic in Clarias gariepinus using lycopene, citric acid, and chlorella.

Despite extensive research on the toxic effects of microplastics (MPs), there is no obtainable data on the use of phytobioremediation against MPs toxicity in fish. This study aimed to investigate the protective role of lycopene, citric acid, and chlorella against the toxic effects of MPs in African catfish (Clarias gariepinus) using hematology, biochemical, antioxidants, erythron profiles (poikilocytosis and nuclear abnormalities) and the accumulation of MPs in tissues as biomarkers. Five groups of fish received: normal diet (control); MPs (500 mg/kg diet) (Group 2); MPs (500 mg/kg diet) + lycopene (500 mg/kg diet) (Group 3); MPs (500 mg/kg diet) + citric acid (30 g/kg diet) (Group 4); and MPs (500 mg/kg diet) + chlorella (50 g/kg diet) (Group 5) for 15 days. Group 2 had significantly higher amounts of MPs in the stomach, gills, and feces, electrolyte imbalances (HCO3, Fe, Na+, K+, Ca+2, Cl-, and anion gap, hematobiochemical alterations, and decreases in the activities of superoxide dismutase, catalase, total antioxidant capacity, and glutathione S-transferases compared to the control group. Additionally, Group 2 had significant increase in the percentage of poikilocytosis, and nuclear abnormalities in RBC's compared to the control group. The co-treatment of MPs-exposed fish with lycopene, citric acid, and chlorella-supplemented diets ameliorated the hematological, biochemical, and erythron profile alterations, but only slightly enhanced the antioxidant activity. Overall, lycopene, citric acid, and chlorella can be recommended as a feed supplement to improve hematobiochemical alterations and oxidative damage induced by MPs toxicity in the African catfish (C. gariepinus).

Identification and Analysis of microRNAs in Chlorella sorokiniana Using High-Throughput Sequencing.

Chlorella is a popular microalga with robust physiological and biochemical characteristics, which can be cultured under various conditions. The exploration of the small RNA content of Chlorella could improve strategies for the enhancement of metabolite production from this microalga. In this study, stress was introduced to the Chlorella sorokiniana culture to produce high-value metabolites such as carotenoids and phenolic content. The small RNA transcriptome of C. sorokiniana was sequenced, focusing on microRNA (miRNA) content. From the analysis, 98 miRNAs were identified in cultures subjected to normal and stress conditions. The functional analysis result showed that the miRNA targets found were most often involved in the biosynthesis of secondary metabolites, followed by protein metabolism, cell cycle, and porphyrin and chlorophyll metabolism. Furthermore, the biosynthesis of secondary metabolites such as carotenoids, terpenoids, and lipids was found mostly in stress conditions. These results may help to improve our understanding of regulatory mechanisms of miRNA in the biological and metabolic process of Chlorella species. It is important and timely to determine the true potential of this microalga species and to support the potential for genetic engineering of microalgae as they receive increasing focus for their development as an alternative source of biofuel, food, and health supplements.

Effects of Nitrogen Supplementation Status on CO2 Biofixation and Biofuel Production of the Promising Microalga Chlorella sp. ABC-001.

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 CO2 biofixation should also be explored. Therefore, in this study, the CO2 biofixation and lipid/carbohydrate production potential of Chlorella sp. ABC-001 were examined under various nitrogen concentrations. The highest biomass productivity and CO2 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 CO2 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.

New insights on improved growth and biogas production potential of Chlorella pyrenoidosa through intermittent iron oxide nanoparticle supplementation.

In the present work, the effect of α-Fe2O3-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 (CODth) as well as theoretical and stoichiometric methane potential (TMP, and SMP) were also estimated. The CODth, 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 VSfed) 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 × 104 m3 ha-1 year-1) compared to control. These findings reveal the potential of IONPs in improving microalgal biogas production.

Microalgae Cultivation Using Screened Liquid Dairy Manure Applying Different Folds of Dilution: Nutrient Reduction Analysis with Emphasis on Phosphorus Removal.

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. NH4-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.

Effect of iron and magnesium addition on population dynamics and high value product of microalgae grown in anaerobic liquid digestate.

In this study, FeSO4 supplementation ranging from 0 to 4.5 mM, and MgSO4 supplementation ranging from 0 to 5.1 mM were investigated to observe the effect on the population dynamics, biochemical composition and fatty acid content of mixed microalgae grown in Anaerobic Liquid Digestate (ALD). Overall, 3.1 mM FeSO4 addition into ALD increased the total protein content 60% and led to highest biomass (1.56 g L-1) and chlorophyll-a amount (18.7 mg L-1) produced. Meanwhile, 0.4 mM MgSO4 addition increased the total carotenoid amount 2.2 folds and slightly increased the biomass amount. According to the microbial community analysis, Diphylleia rotans, Synechocystis PCC-6803 and Chlorella sorokiniana were identified as mostly detected species after confirmation with 4 different markers. The abundance of Chlorella sorokiniana and Synechocystis PCC-6803 increased almost 2 folds both in iron and magnesium addition. On the other hand, the dominancy of Diphylleia rotans was not affected by iron addition while drastically decreased (95%) with magnesium addition. This study helps to understand how the dynamics of symbiotic life changes if macro elements are added to the ALD and reveal that microalgae can adapt to adverse environmental conditions by fostering the diversity with a positive effect on high value product.

Selenium Incorporation to Amino Acids in Chlorella Cultures Grown in Phototrophic and Heterotrophic Regimes.

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.

Enhancing microalga Chlorella sorokiniana CY-1 biomass and lipid production in palm oil mill effluent (POME) using novel-designed photobioreactor.

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 CO2 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.

Sensory properties of aqueous dispersions of protein-rich extracts from Chlorella protothecoides at neutral and acidic pH.

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.

Optimization of heterotrophic cultivation of Chlorella sp. HS2 using screening, statistical assessment, and validation.

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.

Sequencing and comparative analysis of three Chlorella genomes provide insights into strain-specific adaptation to wastewater.

Microalgal Chlorella has been demonstrated to process wastewater efficiently from piggery industry, yet optimization through genetic engineering of such a bio-treatment is currently challenging, largely due to the limited data and knowledge in genomics. In this study, we first investigated the differential growth rates among three wastewater-processing Chlorella strains: Chlorella sorokiniana BD09, Chlorella sorokiniana BD08 and Chlorella sp. Dachan, and the previously published Chlorella sorokiniana UTEX 1602, showing us that BD09 maintains the best tolerance in synthetic wastewater. We then performed genome sequencing and analysis, resulting in a high-quality assembly for each genome with scaffold N50 > 2 Mb and genomic completeness ≥91%, as well as genome annotation with 9,668, 10,240, 9,821 high-confidence gene models predicted for BD09, BD08, and Dachan, respectively. Comparative genomics study unravels that metabolic pathways, which are involved in nitrogen and phosphorus assimilation, were enriched in the faster-growing strains. We found that gene structural variation and genomic rearrangement might contribute to differential capabilities in wastewater tolerance among the strains, as indicated by gene copy number variation, domain reshuffling of orthologs involved, as well as a ~1 Mb-length chromosomal inversion we observed in BD08 and Dachan. In addition, we speculated that an associated bacterium, Microbacterium chocolatum, which was identified within Dachan, play a possible role in synergizing nutrient removal. Our three newly sequenced Chlorella genomes provide a fundamental foundation to understand the molecular basis of abiotic stress tolerance in wastewater treatment, which is essential for future genetic engineering and strain improvement.

Effect of light quality on growth rate, carbohydrate accumulation, fatty acid profile and lutein biosynthesis of Chlorella sp. AE10.

Microalgae are feedstocks for multiple product development based on algal biorefinery concept. The effects of light quality (white, red and blue light emitting diodes) and macro-element starvations on Chlorella sp. AE10 were investigated under 20% CO2 and 850 µmol m-2 d-1. Nitrogen and phosphorus starvations had negative effects on its growth rate. The biomass productivities were decreased from day 1 and the highest one was 1.90 g L-1 d-1 under white light conditions. Phosphorus starvation promoted carbohydrate accumulation under three LED light sources conditions and the highest carbohydrate content was 75.9% using red light. Blue light increased lutein content to 9.58 mg g-1. The content of saturated fatty acids was significantly increased from 37.51% under blue light and full culture medium conditions to 77.44% under blue light and nitrogen starvation conditions. Chlorella sp. AE10 was a good candidate for carbohydrate and lutein productions.

Growth optimization, free radical scavenging and antibacterial potential of Chlorella sp. SRD3 extracts against clinical isolates.

AIM: The aim of present work was to explore the potential of Chlorella sp. SRD3 extracts for antioxidant and antibacterial activity along with the evaluation of minimum inhibitory concentration (MIC) and haemolytic activity to detect RBC cell damage. METHODS AND RESULTS: Screening and isolation of microalgae was performed using bold basal medium under normal illuminance (at 27°C) and microscopic observation. Growth of the microalgae was optimized using a different medium and light source. The isolated microalgae incubated under fluorescent light when cultured in F/2 medium showed a highest dry biomass yield of 3·77 ± 0·1 g l-1 , when compared to the growth under direct sunlight (2·74 ± 0·07 g dwt l-1 ). The quantitative analysis of extracts revealed higher phenols, flavonoids and proanthocyanidins in ethyl acetate and hexane extracts followed by methanol. The antioxidant activity of extracts was tested against 1-diphenyl-2-picrylhydrazyl and ABTS radical, its reducing power assay was performed. From antibacterial activity, the two extracts showed better inhibition against Gram-negative bacteria. Also, they resulted in very low MIC values with effective activity against pathogens. In haemolytic activity, no haemolysis occurred, when the concentration (µg ml-1 ) was below 64 for methanol and 32 for ethyl acetate extract. In addition, Chlorella sp. extracts were characterized by GC-MS analysis to detect the major compounds. CONCLUSION: The polar extracts revealed satisfactory results against the clinical isolates and the compounds responsible were reflected in the GC-MS spectrum. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study revealed significant biological potentials of the green alga, Chlorella sp. such as antioxidant, antibacterial and hemolytic activities. Therefore, this vital source might serve as a cost-effective, alternative choice to the pharmaceutical and food industries in the near future.

Effects of various abiotic factors on biomass growth and lipid yield of Chlorella minutissima for sustainable 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.

Excessive phosphorus caused inhibition and cell damage during heterotrophic growth of Chlorella regularis.

A high phosphorus concentration is widely accepted as favorable for enhancing both microalgae growth and lipid accumulation; however, excessively high P could be counter-productive. In this study, we investigated the effects of increasing P levels (5.4, 25, 45, 150, and 250 mg-P L-1) on the heterotrophic cultivation of Chlorella regularis. Microalgae growth was inhibited and cells were severely damaged in response to highly excessive P levels (≥150 mg-P L-1). In particular, 250 mg-P L-1 resulted in a ∼40% decrease in cell density and a ∼70% loss of cell viability. Microalgae damage induced by excessive phosphorus included enlarged cell size, deformation of cell walls, and disorganization of organelles. These negative effects were associated with the over-accumulation of polyphosphates within cells, which may further cause binding of P to intracellular components. Although P is an essential nutrient, excessive P lowers cell growth and viability.

Cultivation of microalgae Chlorella zofingiensis on municipal wastewater and biogas slurry towards bioenergy.

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.

Influence of Water Depth on Microalgal Production, Biomass Harvest, and Energy Consumption in High Rate Algal Pond Using Municipal Wastewater.

The high rate algal ponds (HRAP) powered and mixed by a paddlewheel have been widely used for over 50 years to culture microalgae for the production of various products. Since light incidence is limited to the surface, water depth can affect microalgal growth in HRAP. To investigate the effect of water depth on microalgal growth, a mixed microalgal culture constituting three major strains of microalgae including Chlorella sp., Scenedesmus sp., and Stigeoclonium sp. (CSS), was grown at different water depths (20, 30, and 40 cm) in the HRAP, respectively. The HRAP with 20cm of water depth had about 38% higher biomass productivity per unit area (6.16 ± 0.33 g·m⁻²·d⁻¹) and required lower nutrients and energy consumption than the other water depths. Specifically, the algal biomass of HRAP under 20c m of water depth had higher settleability through larger floc size (83.6% settleability within 5 min). These results indicate that water depth can affect the harvesting process as well as cultivation of microalgae. Therefore, we conclude that water depth is an important parameter in HRAP design for mass cultivation of microalgae.

Effects of Selenite on Unicellular Green Microalga Chlorella pyrenoidosa: Bioaccumulation of Selenium, Enhancement of Photosynthetic Pigments, and Amino Acid Production.

Microalgae were studied as function bioaccumulators of selenium (Se) for food and feed supplement. To investigate the bioaccumulation of Se and its effects on the unicellular green alga Chlorella pyrenoidosa, the algal growth curve, fluorescence parameters, antioxidant enzyme activity, and fatty acid and amino acid profiles were examined. We found that Se at low concentrations (≤40 mg L-1) positively promoted algal growth and inhibited lipid peroxidation and intracellular reactive oxygen species. The antioxidative effect was associated with an increase in the levels of glutathione peroxidase, catalase, linolenic acid, and photosynthetic pigments. Meanwhile, a significant increase in amino acid and organic Se content was also detected in the microalgae. In contrast, we found opposite effects in C. pyrenoidosa exposed to >60 mg L-1 Se. The antioxidation and toxicity appeared to be correlated with the bioaccumulation of excess Se. These results provide a better understanding of the effect of Se on green microalgae, which may help in the development of new technological applications for the production of Se-enriched biomass from microalgae.

Optimal Temperature and Light Intensity for Improved Mixotrophic Metabolism of Chlorella sorokiniana Treating Livestock Wastewater.

Mixotrophic microalgal growth gives a great premise for wastewater treatment based on photoautotrophic nutrient utilization and heterotrophic organic removal while producing renewable biomass. There remains a need for a control strategy to enrich them in a photobioreactor. This study performed a series of batch experiments using a mixotroph, Chlorella sorokiniana, to characterize optimal guidelines of mixotrophic growth based on a statistical design of the experiment. Using a central composite design, this study evaluated how temperature and light irradiance are associated with CO2 capture and organic carbon respiration through biomass production and ammonia removal kinetics. By conducting regressions on the experimental data, response surfaces were created to suggest proper ranges of temperature and light irradiance that mixotrophs can beneficially use as two types of energy sources. The results identified that efficient mixotrophic metabolism of Chlorella sorokiniana for organics and inorganics occurs at the temperature of 30-40°C and diurnal light condition of 150-200 µmol E·m2·s-1. The optimal specific growth rate and ammonia removal rate were recorded as 0.51/d and 0.56/h on average, respectively, and the confirmation test verified that the organic removal rate was 105 mg COD·l-1·d-1. These results support the development of a viable option for sustainable treatment and effluent quality management of problematic livestock wastewater.