Influence of Nitrogen to Phosphorus Ratio and CO2 Concentration on Lipids Accumulation of Scenedesmus dimorphus for Bioenergy Production and CO2 Biofixation.

The potential of Scenedesmus dimorphus microalgae for CO2 biofixation and lipid biosynthesis for bioenergy applications was evaluated in this study. Batch experiments were conducted using synthetic tertiary municipal wastewater samples at several nitrogen to phosphorus (NP) ratios (1 : 1 to 8 : 1) and CO2 concentrations (∼0%, 2%, 4%, and 6% CO2 in supplied air). Scenedesmus dimorphus was cultivated for 25 days and the growth is highly dependent on the CO2 concentration and the NP ratio. An NP ratio of 2 : 1 produces a biomass yield of 733 mg/L when the microalga culture was supplied with air enriched with 2% CO2 . The maximum CO2 biofixation rate of 49.6 mg L-1 d-1 is at an NP ratio of 8 : 1 with 4% CO2 . A colorimetric technique depending on sulpho-phospho-vanillin (SPV) was utilized for the determination of the intracellular lipid content. The highest lipid content of 31.6% as the dry weight of the biomass is at an NP ratio of 1 : 1 and 6% CO2 . These results indicate that supplementation of suitable CO2 with favorable NP ratio has a considerable effect on lipid accumulation in the microalgae Scenedesmus dimorphus biomass.

The relationship between sulfur metabolism and tolerance of hexavalent chromium in Scenedesmus acutus (Spheropleales): Role of ATP sulfurylase.

Sulfur availability and the end products of its metabolism, cysteine, glutathione and phytochelatins, play an important role in heavy metal tolerance, chromium included. Sulfate and chromate not only compete for the transporters but also for assimilation enzymes and chromium tolerance in various organisms has been associated to differences in this pathway. We investigated the mechanisms of Cr(VI)-tolerance increase induced by S-starvation focusing on the role of ATP sulfurylase (ATS) in two strains of Scenedesmus acutus with different chromium sensitivity. S-starvation enhances the defence potential by increasing sulfate uptake/assimilation and decreasing chromium uptake, thus suggesting a change in the transport system. We isolated two isoforms of the enzyme, SaATS1 and SaATS2, with different sensitivity to sulfur availability, and analysed them in S-sufficient and S-replete condition both in standard and in chromium supplemented medium. SaATS2 expression is different in the two strains and presumably marks a different sulfur perception/exploitation in the Cr-tolerant. Its induction and silencing are compatible with a role in the transient tolerance increase induced by S-starvation. This enzyme can however hardly be responsible for the large cysteine production of the Cr-tolerant strain after starvation, suggesting that cytosolic rather than chloroplastic cysteine production is differently regulated in the two strains.

Morphological characterization, growth appraisal, and probing biofuels potential of newly isolated Scenedesmus sp. from desert Cholistan.

Microalgae have an excellent potential for producing valuable natural products, including biofuels. Therefore, it is imperative to explore and document the existing microalgal flora and utilize their potentials to cope the increasing human needs. The present work aims at exploring and characterizing newly isolated microalgae from desert Cholistan, a habitat with myriad algal diversity. Light microscopy, scanning electron microscopy, and molecular phylogenetic approaches were used for species-level identification. Characterization and growth optimization of Scendesmus sp. were analyzed under three different growth modes to determine the most favorable conditions for increasing biomass, growth rate, and lipid content. The results revealed that mixotrophic (MT) mode significantly increases photosynthetic activity, growth rate, and lipid content with glycerol as supplement carbon source. The investigated Scenedesmus dimorphous produced a maximum dry weight of 1.73 g L-1 , improved fatty acid methyl esters profile and yield lipid up to 40% of DCW (68 g L-1 ) under MT mode, which is almost double to that of photoautotrophic cultivation. The glycerol availability in medium has been identified as the critical element for boosting growth and lipid content. Thus, it can reduce the cost of biofuel production.

Scenedesnus rotundus isolated from the petroleum effluent employs alternate mechanisms of tolerance to elevated levels of Cadmium and Zinc.

Scenedesmus rotundus was isolated from metal contaminated petroleum industry effluent and its tolerance to Cadmium and Zinc was tested using different concentrations of CdCl2 and ZnCl2 ranging from 0.001 mM to 1.0 mM of Cd and 0.03 mM to 1.21 mM of Zn amended in Bolds Basal medium. The changes in cell count recorded at regular intervals upto a period of 24 days revealed a concentration dependent inhibition in growth. Concentration of the metal, at which 50% of the cells are live and metabolically active referred to as EC50 was calculated as 0.04 mM for Cd and 0.2 mM for Zn. Further, the effect of EC50 of the metals on the protein content, uptake of metals at varying pH, oxidative stress markers including lipid peroxidation, protein oxidation andnd oxygen uptake, levels of enzymatic antioxidants such as catalase and superoxide dismutase and non-enzymatic antioxidants namely, GSH and PC4 were determined. Though a direct correlation could not be drawn between pH and metal uptake, the compartmentalization of the metal during the lag phase and exponential phase was evident, most of the metal was present in extracellular fractions in the former, while in the later it was internalized. Our study shows a clear correlation between toxicity of Cd and the ability of the algae to synthesize PC4 from GSH and chelate it leading to detoxification, while Zn treatment led to an increase in the activity of catalase and superoxide dismutase and replete GSH pools. Further the changes in the cell wall structure at EC50 of Cd and Zn were studied. This is the first report on effect of heavy metals on the structural modifications of the cell wall of Scenedesmus in general and Scenedesmus rotundus in particular, indicating appearance of granules on the entire cell surface in both Cd and Zn treatments, with the degree of granulation increasing in the order of pH 12 > 10 > 8 in Cd treatment. Further structures of higher order resembling minute wheels are observed in Cd treated cells are also reported.

Comparison of monoculture and mixed culture (Scenedesmus obliquus and wild algae) for C, N, and P removal and lipid production.

This study compared the efficiency of nutrient removal and lipid accumulation by a monoculture of Scenedesmus obliquus and mixed cultures of microalgae. The highest removal efficiencies of ammonium (99.2%), phosphate (91.2%), and total organic carbon (83.6%) occurred in the monoculture. All the mixed cultures were dominated by S. quadricauda; in some mixed cultures, the proportions of Chlamydomonas reinhardtii and C. microsphaera reached > 20%. The lipid content and lipid production in the monoculture were 15.9% and 52.3 mg kg-1, respectively, significantly higher than those in all the mixed cultures of microalgae. In all the mixed cultures, the proportion of palmitic acid was > 50%. The results suggest that the monoculture had advantages over the mixed culture of microalgae in terms of nutrient removal and lipid production.

Influences of carbon and nitrogen sources and metal ions on the heterotrophic culture of Scenedesmus sp. LX1.

In this study, the influences of organic carbon sources (OCS, including xylose, glucose, maltose, sucrose, and starch) and inorganic and organic nitrogen sources (INS, including ammonia chloride and sodium nitrate; ONS, including arginine, alanine, proline, and valine) and metal ions (including Na+, K+, Mn2+, Zn2+ and Cu2+) on the growth, lipid accumulation, and nitrogen and phosphorus (N&P) removal capabilities of oleaginous Scenedesmus sp. LX1 under heterotrophic conditions were investigated. The results showed that glucose was the only OCS for Scenedesmus sp. LX1 to grow well with specific growth rate of 0.935 days-1, maximum biomass of 1.72 g L-1, and largest removal rates of N&P and organic carbon reaching 72.228%, 93.034%, and 19.208%, respectively. After 11 days of cultivation, the maximal biomass reached in the group with starch or glucose while maximal lipid and triacylglycerol (TAG) yields reached in the groups with maltose and sucrose, respectively. Sodium nitrate was best nitrogen source as the largest algal density, maximal yields of lipids and TAGs, and highest N&P removal rates reached up to 1.105 × 107 cells·mL-1, 196.70 mg L-1, 5.19 mg L-1, 89.61% and 100%, respectively. Scenedesmus sp. LX1 was found to have great tolerance to Na+, K+, Mn2+, and Zn2+ while 0.5 mg L-1 Cu2+ had a strong inhibition on growth and N&P removal rate of Scenedesmus sp. LX1. Concentration increasing of five metal ions all caused the yield increases of microalgal lipid and TAGs. Graphical abstract.

Effects of aqueous extracts from the rhizome of Pontederia cordata on the growth and interspecific competition of two algal species.

Single and co-culture systems of Microcystis aeruginosa and Scenedesmus obliquus were prepared with different initial algal densities and treated with different concentrations of aqueous extracts from the rhizome of Pontederia cordata to study its inhibitory effect on algal growth and the competitive relationship between these two algal species. The results showed that aqueous extracts could inhibit the growth of M. aeruginosa and S. obliquus, and the inhibition rate of aqueous extracts on the growth of M. aeruginosa was always higher than that of S. obliquus. A Lotka-Volterra competition model revealed that these two algal species can co-exist without the addition of aqueous extracts, and S. obliquus exhibited a stronger ability to compete than that of M. aeruginosa. Meanwhile, the dominant algal species changed with the addition of aqueous extracts regardless of the initial ratios of the two algae. The species ratio of mixed cultures had a strong effect on the interspecific interaction between the two algae. The higher proportion of S. obliquus in the initial proportion of two algae, the stronger competitive ability of S. obliquus when compared with that of M. aeruginosa.

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.

The growth of Scenedesmus quadricauda in RO concentrate and the impacts on refractory organic matter, Escherichia coli, and trace organic compounds.

This study achieves a better operational simplicity for the phycoremediation of reverse osmosis (RO) concentrate using Scenedesmus quadricauda microalgae. Under continuous illumination with CO2 supplementation, algal growth in the RO concentrate resulted in a conversion of polymeric organic matter (a mixture of humic substances and polysaccharides) to biodegradable fractions and their prompt removal along with inorganic nutrients (NO3- and PO43-). The algal-induced degradation of humic-like substances which are typically refractory to microbial decomposition was demonstrated in an indirect manner. In this study, we also investigated the effects of algal treatment on the growth of Escherichia coli and removal of trace organic compounds (TOrCs) from the RO concentrate. Our results indicate that algal treatment of the RO concentrate using aeration with 10% (v/v) CO2 under continuous illumination is highly feasible as a safe and inexpensive technology to remove non- or slowly-biodegradable organic matter, reduce enteric bacteria, and attenuate TOrCs in wastewater. However, the results should not be generalized, but critically discussed, due to limitations of using the synthetic RO concentrate in evaluating the performance of wastewater remediation with microalgae.

Growth modeling of the green microalga Scenedesmus obliquus in a hybrid photobioreactor as a practical tool to understand both physical and biochemical phenomena in play during algae cultivation.

In recent years, numerous studies have justified the use of microalgae as a sustainable alternative for the generation of different types of fuels, food supplementation, and cosmetics, as well as bioremediation processes. To improve the cost/benefit ratio of microalgae mass production, many culture systems have been built and upgraded. Mathematical modeling the growth of different species in different systems has become an efficient and practical tool to understand both physical and biochemical phenomena in play during algae cultivation. In addition, growth modeling can guide design changes that lead to process optimization. In the present work, growth of the green microalga Scenedesmus obliquus was modeled in a hybrid photobioreactor that combines the characteristics of tubular photobioreactors (TPB) with thin-layer cascades (TLC). The system showed productivity greater than 8.0 g m-2 day-1 (dry mass) for CO2 -fed cultures, and the model proved to be an accurate representation of experimental data with R2 greater than 0.7 for all cases under variable conditions of temperature and irradiance to determine subsystem efficiency. Growth modeling also allowed growth prediction relative to the operating conditions of TLC, making it useful for estimating the system given other irradiance and temperature conditions, as well as other microalgae species.

Carbon-dioxide biofixation and phycoremediation of municipal wastewater using Chlorella vulgaris and Scenedesmus obliquus.

The pure cultures of microalgae Chlorella vulgaris ATCC 13482 and Scenedesmus obliquus FACHB 417 were grown in municipal wastewater in 7-L airlift bubble column photobioreactor supplied with 5% CO2/air (v/v). Batch experiments were conducted at 25 °C with 14-h light/10-h dark cycle for a period of 10 days. The CO2 capture efficiencies for both the microalgae were monitored in terms of their respective biomass productivities, carbon contents, and CO2 consumption rates. In the present study, the initial concentration of ammonia (43.7 mg L-1) was decreased to 2.9 and 3.7 mg L-1 by C. vulgaris and S. obliquus, respectively. And, the initial concentration of phosphate (18.5 mg L-1) was decreased to 1.1 and 1.6 mg L-1 by C. vulgaris and S. obliquus, respectively. CO2 biofixation rates by C. vulgaris and S. obliquus, cultivated in municipal wastewater, were calculated to be 140.91 and 129.82 mg L-1 day-1, respectively. The findings from the present study highlight the use of microalgae for wastewater treatment along with CO2 uptake and biomass utilization for pilot scale production of biodiesel, biogas, feed supplements for animals, etc., thus minimizing the production costs.

Improvement on lipid production by Scenedesmus obliquus triggered by low dose exposure to nanoparticles.

Carbon nanotubes (CNTs), α-Fe2O3 nanoparticles (nano Fe2O3) and MgO nanoparticles (nano MgO) were evaluated for the effects on algae growth and lipid production. Nano Fe2O3 promoted cell growth in the range of 0-20 mg·L-1. CNTs, nano Fe2O3 and nano MgO inhibited cell growth of Scenedesmus obliquus at 10, 40 and 0.8 mg·L-1 respectively. Neutral lipid and total lipid content increased with the increasing concentration of all tested nanoparticles. The maximum lipid productivity of cultures exposed to CNTs, nano Fe2O3 and nano MgO was observed at 5 mg·L-1, 5 mg·L-1 and 40 mg·L-1, with the improvement by 8.9%, 39.6% and 18.5%. High dose exposure to nanoparticles limited increase in lipid productivity, possibly due to the repression on cell growth caused by nanoparticles-catalyzed reactive oxygen species (ROS) generation, finally leading to reduction in biomass and lipid production. Reduced accumulation of fatty acids of C18:3n3, C18:3n6 and C20:2 was observed in cells exposed to nanoparticles.

Enrichment of highly settleable microalgal consortia in mixed cultures for effluent polishing and low-cost biomass production.

Microalgae cultivation is a promising technology for integrated effluent polishing and biofuel production, but poor separability of microalgal cells hinders its industrial application. This study intended to selectively enrich settleable microalgal consortia in mixed culture by applying "wash-out" pressure, which was realized by controlling settling time (ST) and volume exchange ratio (VER) in photo-SBRs. The results demonstrated that highly settleable microalgal consortia (settling efficiency>97%; SVI = 17-50 mL/g) could be enriched from indigenous algal cultures developed in WWTP's effluent. High VER was the key factor for the fast development of settleable microalgae. VER was also a controlling factor of the algal community structure. High VERs (0.5 and 0.7) resulted in the dominance of diatom, while low VER (0.2) facilitated the dominance of cyanobacteria. The settleable microalgal consortia were very efficient in phosphorus removal (effluent PO43--P<0.1 mg/L; removal efficiency>99%), which was largely attributed to intensive chemical precipitation of phosphate induced by high pH (8.5-10). However, the high pH decreased the bioavailable inorganic carbon, resulting in incomplete nitrate removal (effluent NO3--N = 2.2-4 mg/L; removal efficiency = 61-79%) under high VERs and low lipid content (up to 10%) in the settleable microalgae. This problem could be resolved by sparging CO2 or controlling pH. Overall, this study demonstrated a simple and effective method to overcome the separation challenge in scale-up of microalgae biotechnology for advanced wastewater purification and biofuel production.

Screening of microalgae for integral biogas slurry nutrient removal and biogas upgrading by different microalgae cultivation technology.

The microalgae-based technology has been developed to reduce biogas slurry nutrients and upgrade biogas simultaneously. In this work, five microalgal strains named Chlorella vulgaris, Scenedesmus obliquus, Selenastrum capricornutum, Nitzschia palea, and Anabaena spiroides under mono- and co-cultivation were used for biogas upgrading. Optimum biogas slurry nutrient reduction could be achieved by co-cultivating microalgae (Chlorella vulgaris, Scenedesmus obliquus, and Nitzschia palea) with fungi using the pelletization technology. In addition, the effects of different ratio of mixed LED light wavelengths applying mixed light-emitting diode during algae strains and fungi co-cultivation on CO2 and biogas slurry nutrient removal efficiency were also investigated. The results showed that the COD (chemical oxygen demand), TN (total nitrogen), and TP (total phosphorus) removal efficiency were 85.82 ± 5.37%, 83.31 ± 4.72%, and 84.26 ± 5.58%, respectively at red: blue = 5:5 under the co-cultivation of S. obliquus and fungi. In terms of biogas upgrading, CH4 contents were higher than 90% (v/v) for all strains, except the co-cultivation with S. obliquus and fungi at red: blue = 3:7. The results indicated that co-cultivation of microalgae with fungi under mixed light wavelengths treatments was most successful in nutrient removal from wastewater and biogas upgrading.

Enhanced attached growth of microalgae Scenedesmus. LX1 through ambient bacterial pre-coating of cotton fiber carriers.

The role of bacteria/extracellular polymeric substances (EPS) coated carriers on attached microalgae growth in suspended-solid phase photobioreactor (sspBR) was assessed in this study. The results showed that pre-coating cotton with ambient bacteria and their EPS improved the attached microalgal growth by as much as 230% in terms of attached microalgae density. Additionally, the single cell dry weight, chemical composition and oxygen evolving activity of attached microalgae were significantly affected by the presence of bacteria/EPS coating on the cotton carriers. The protein content of microalgae cells cultivated in the ssPBRs with carriers coated by bacteria and sterilized bacteria were on average 26% and 15% more than uncoated carriers, respectively. Through absorbing and immobilizing nutrients from the bulk medium, the bacteria/EPS coating provided the attached microalgae with nitrogen/phosphorus for protein synthesis, especially during the late stages of batch cultivation.

Microalgae harvesting by pH adjusted coagulation-flocculation, recycling of the coagulant and the growth media.

Coagulation-flocculation can be considered as one of the least energy intensive microalgae biomass harvesting processes. However, cost of the coagulant and biomass contamination are two critical issues that need to be considered. In this study, ferric chloride (72-96mg/L) was used to effectively harvest Scenedesmus sp. (530mg/L) - grown in BG-11 media and wastewater. Reducing the culture pH below 6.5, greatly improved the harvesting efficiency. Acidic solution (pH 1.0) was very effective to recover (almost 90%) the associated iron from the harvested biomass. Scenedesmus sp. was able to grow in the supernatant and utilize the residual iron in it. Iron extracted solution, with a supplementation of 9.8mg/L ferric chloride, was able to achieve similar harvesting efficiency. The potential recovery of iron from the harvested biomass and its reuse in the harvesting can improve the biomass quality for subsequent downstream processing while reducing the cost.

Removal of nutrients and organic pollution load from pulp and paper mill effluent by microalgae in outdoor open pond.

A mixed culture of microalgae, containing two Scenedesmus species, was analysed to determine its potential in coupling of pulp and paper mill effluent treatment and microalgal cultivation. Laboratory studies suggested that 60% concentration of wastewater was optimum for microalgal cultivation. A maximum of 82% and 75% removal of BOD and COD respectively was achieved with microalgal cultivation in outdoor open pond. By the end of the cultivation period, 65% removal of NO3-N and 71.29% removal of PO4-P was observed. The fatty acid composition of mixed microalgal culture cultivated with effluent showed the palmitic acid, oleic acid, linoleic acid and α-linolenic acid as major fatty acids. The results obtained suggest that pulp and paper mill effluent could be used effectively for cultivation of microalgae to minimise the freshwater and nutrient requirements.

Evaluation of novel thermo-resistant Micractinium and Scenedesmus sp. for efficient biomass and lipid production under different temperature and nutrient regimes.

Despite the vast interest in microalgae as feedstock for biodiesel production, relatively few studies examined their response to diurnal temperature fluctuation. Here, we describe biomass and lipid productivities and fatty acid profiles of thermo-resistant Micractinium sp. and Scenedesmus sp. grown in batch cultures in a laboratory set-up that mimics a typically warm summer day in Central Anatolia with a 16-h light temperature of 30°C and 8-h dark temperature of 16°C (30°C (day)/16°C (night)). Both strains can survive a temperature range of 10-50°C. We found the lipid productivities of Micractinium sp. and Scenedesmus sp. as 30/21mgL(-1)d(-1) and 6/7mgL(-1)d(-1), respectively during the 30°C (day)/16°C (night) cycle. Saturated fatty acid content increased with increasing temperature. Additionally, we cultured Micractinium sp. under Nitrogen (N) and Phosphorus (P) limiting conditions. Highest lipid productivity of 85.4±2mgL(-1)d(-1) was obtained under P-depletion during exponential growth phase. Oleic acid amount also increased eight fold during P-deplete.

Simultaneous nitrogen, phosphorous, and hardness removal from reverse osmosis concentrate by microalgae cultivation.

While reverse osmosis (RO) is a promising technology for wastewater reclamation, RO concentrate (ROC) treatment and disposal are important issues to consider. Conventional chemical and physical treatment methods for ROC present certain limitations, such as relatively low nitrogen and phosphorus removal efficiencies as well as the requirement of an extra process for hardness removal. This study proposes a novel biological approach for simultaneous removal of nitrogen, phosphorus, and calcium (Ca(2+)) and magnesium (Mg(2+)) ions from the ROC of municipal wastewater treatment plants by microalgal cultivation and algal biomass production. Two microalgae strains, Chlorella sp. ZTY4 and Scenedesmus sp. LX1, were used for batch cultivation of 14-16 days. Both strains grew well in ROC with average biomass production of 318.7 mg/L and lipid contents up to 30.6%, and nitrogen and phosphorus could be effectively removed with efficiencies of up to 89.8% and 92.7%, respectively. Approximately 55.9%-83.7% Ca(2+) could be removed from the system using the cultured strains. Mg(2+) removal began when Ca(2+) precipitation ceased, and the removal efficiency of the ion could reach up to 56.0%. The most decisive factor influencing Ca(2+) and Mg(2+) removal was chemical precipitation with increases in pH caused by algal growth. The results of this study provide a new biological approach for removing nitrogen, phosphorous, and hardness from ROC. The results suggest that microalgal cultivation presents new opportunities for applying an algal process to ROC treatment. The proposed approach serves dual purposes of nutrient and hardness reduction and production of lipid rich micro-algal biomass.

Cultivation of freshwater microalga Scenedesmus sp. using a low-cost inorganic fertilizer for enhanced biomass and lipid yield.

The potential of an inorganic fertilizer as an alternative nutrient source for the cultivation of Scenedesmus sp. IMMTCC-6 was investigated. With a preliminary study at a shake-flask scale, the microalgae cultivation was scaled up in a photobioreactor containing an inorganic fertilizer medium. Microalgae cultured in a shake flask containing 0.1 g L(-1) of urea and 1.0 g L(-1) of NPK (Nitrogen: Phosphorus: Potassium) fertilizers showed a promising result in biomass productivity. During the scale-up study in a photobioreactor the specific growth rate (µ d(-1)), biomass yield (g L(-1)), and total biomass productivity (mg L(-1) d(-1)), was found to be 0.265, 1.19 and 66.1, respectively. The lipid yield (%) as per dry cell weight (DCW) and lipid productivity (mg L(-1) d(-1)) was found to be a maximum of 28.55 and 18.87, respectively, in a stationary phase of the microalgae growth. The fatty acids methyl ester profile was proven to be desirable for biodiesel production.