Production of microalgae from olive mill wastewater

With olive mill wastewater (alpechin) used as a nutrient medium, the influence of the aeration level and the composition of the culture medium were analysed in relation to the concentration of alpechin and KNO₃ added in a batch culture of the microalgae Chlorella pyrenoidosa and Scenedesmus obliquus. The pH and temperature of the culture were previously fixed at 6.5 and 30°C. As kinetic parameters, the maximum specific growth rate (μ_m) and the biomass productivity (P_max) were determined. The biomass composition obtained was evaluated for chlorophyll and crude protein content as well as fatty-acid composition corresponding to the lipid fraction. To achieve a balanced biomass composition in terms of proteins and lipids, the most suitable conditions included an alpechin concentration of 10%, without the addition of nitrates and with an aeration level of 1 v/v min. Under these conditions, the μ_m values reached with both microalgae were around 0.03h⁻¹.     

Harvesting Chlorella species using magnetic iron oxide nanoparticles

Microalgae are very useful organisms as it provides many beneficial products for human use. For large scale cultivation and further applications, its harvesting procedure needs to be enhanced to make the production process of the end product highly affordable. Magnetic nanoparticles have great potential to harvest microalgae as it can easily attract and attach to the algal cell surface forming a layer, which can be harvested quickly under the influence of a magnetic field. Our work on Chlorella pyrenoidosa and Chlorella minutissima shows, 500 mg of the synthesized bare iron oxide nanoparticles, harvests 90% of Chlorella pyrenoidosa (1 g L^?1), in 60 s at pH 3 and 600 mg iron oxide nanoparticles, harvests 85% of Chlorella minutissima (1 g L^?1) in 60 s at pH 5, which can decrease the amount of time and energy consumed in the overall production costs.     

Indole-3-acetic acid in the culture medium of two axenic green microalgae

In the view of the facts that algal extracts have been used in agriculture asa source of plant growth stimulating agents and IAA has been shown to bepresent in the extracts, a study was planned to establish whether or notaxenic algae can produce IAA. Evidence is provided for extracellular IAAproduction during culture of two axenic green microalgae. IAAidentification was based on co-chromatography with the standard, analysisof UV and fluorescent spectra, and gas chromatography – selectedion-monitoring mass spectrometry. HPLC analyses showed that underthe experimental conditions the amounts of IAA released to the mediumby Scenedesmus armatus and Chlorella pyrenoidosa weregenerally low. IAA tended to occur in Scenedesmus armatus culturemedium at higher concentrations than in that of Chlorellapyrenoidosa. In fast-growing cultures of Scenedesmus armatus,constantly aerated with CO₂/air mixture, the concentration of IAAcalculated per cell was less than in the slow-growing cultures.     

自养和兼养条件下蛋白核小球藻昼夜节律的响应

【目的】研究自养和兼养两种培养方式对蛋白核小球藻(Chlorella pyrenoidosa)生长、细胞分裂和生化组分积累的影响,探讨人工培养蛋白核小球藻的昼夜节律响应机制和优化技术。【方法】小球藻自养培养采用BG11培养基,兼养培养基在BG11培养基中添加4种不同浓度(1、5、10、20 g/L)的葡萄糖,培养周期为10 d。血球板计数法测定藻细胞浓度,干重法测定藻细胞生物量。显微观察藻细胞大小和分裂情况。脂染色法测定小球藻总脂的含量,藻细胞的叶绿素、蛋白和淀粉分别采用甲醇、氢氧化钠、硝酸钙浸提后通过紫外分光光度法定量测定。【结果】葡萄糖兼养培养对蛋白核小球藻具有显著的促生长效应,最适浓度为10 g/L。10 d收获时,兼养组(10 g/L葡萄糖)藻细胞浓度和干重分别是自养组的2.57倍和6.73倍。分析一昼夜中的藻细胞增殖规律可知,第2天和第5天时自养组中增殖的新生子细胞约有76.00%在黑暗期分裂产生,而兼养组中第2天和第5天光照期的新细胞增殖量占比分别达到40.90%和67.50%。一昼夜内藻细胞大小的迁移动态监测表明,第2天自养组藻细胞的体积变化静息期为8 h,兼养组只有4 h;第5天两组藻细胞大小迁移动态的昼夜节律明显,但兼养组黑暗结束后较大细胞(D6μm)占比显著高于自养组。第8天时,兼养组藻细胞已处于稳定期,总脂和蛋白含量均显著高于自养组,藻细胞总脂和色素含量在一昼夜中相对稳定,但蛋白和淀粉含量分别在光照8 h和12 h左右达到峰值。从第2天开始,对兼养组细胞每天进行2 h光延长,收获时藻细胞浓度和干重分别比对照组提高13%和11%。【结论】葡萄糖兼养培养能大幅提高蛋白核小球藻的生物量。蛋白核小球藻生长增殖与生化组分积累均受昼夜节律调控,自养条件下藻细胞以光照期生长黑暗期增殖为主。兼养培养提高藻细胞生物量的机制在于缩短藻细胞生长静息期,在昼夜节律中加速藻细胞生长并显著提高通过细胞周期检查点的细胞比例,光照期效应尤其明显。藻细胞蛋白和淀粉含量昼夜节律明显,最佳收获时间分别在光照8 h和12 h后。     

Cultivation of a new microalga,Micractinium reisseri,in municipal wastewater for nutrient removal,biomass, lipid,and fatty acid production

Coupling of advanced wastewater treatment with microalgae cultivation for low-cost lipid production was demonstrated in this study. The microalgal species Micractinium reisseri and Scenedesmus obliquus were isolated from municipal wastewater mixed with agricultural drainage. M. reisseri was selected based on the growth rate and cultivated in municipal wastewater (influent, secondary and tertiary effluents) which varied in nutrient concentration. M. reisseri showed an optimal specific growth rate (μ_opt) of 1.15, 1.04, and 1.01 1/day for the influent and the secondary and tertiary effluents, respectively. Secondary effluent supported the highest phosphorus removal (94%) and saturated fatty acid content (40%). The highest lipid content (40%), unsaturated fatty acid content, including monounsaturated and polyunsaturated fatty acids (66%), and nitrogen removal (80%) were observed for tertiary effluent. Fatty acids accumulating in the microalgal biomass (M. reisseri) were mainly composed of palmitic acid, oleic acid, linoleic acid, and a-linolenic acid. Cultivation of M. reisseri using municipal wastewater served a dual function of nutrient removal and biofuel feedstock generation.     

Bioremediation of synthetic high–chemical oxygen demand wastewater using microalgal species Chlorella pyrenoidosa

The microalgal species Chlorella pyrenoidosa was cultivated in synthetic wastewater of initial chemical oxygen demand (COD), nitrate, and phosphate concentrations of 5000, 100, and 40 mg/L, respectively. The aim of the study was to find out the tolerance of microalgae to different COD concentrations and the extent of COD degradation at those concentrations. Three dilutions of wastewater (initial COD concentrations 5000, 3000, and 1000 mg/L) and three inoculum sizes (0.1, 0.2, and 0.3 g/L) were considered for the study. The experimental parameters such as total organic carbon, total inorganic carbon, COD, optical density, total solids, nitrate, and phosphate were measured on a daily basis. Biodegradation kinetics was determined for all cases using first-order reaction and Monod degradation equations. Optimal results showed that up to 90% reduction in TOC was obtained for 1000 COD wastewater while only 38% reduction in total organic carbon (TOC) was achieved for 5000 COD wastewater. Over 95% reduction in nitrate and nearly 90% removal of phosphate were obtained with the lowest microalgal inoculum concentration (i.e., 0.1 g/L) for all COD dilutions. This study showed that microalgal species C. pyrenoidosa can successfully degrade the organic carbon source (i.e., acetate) with significant removal efficiencies for nitrate and phosphate.     

The potential of microalgal biomass production for biotechnological purposes using wastewater resources

The utilization of microalgae for wastewater treatment represents an attractive opportunity for wastewater valorization through the use of the produced biomass. Five strains of microalgae were isolated from municipal wastewater and grown in autoclaved and non-autoclaved effluent at 30 °C and 150 μmol photons m^?2 s^?1 to study biomass production, nutrient removal, and the biochemical composition of the biomass. All strains reached high biomass productivity (35.6 to 54.2 mg dry weight L^?1 day^?1) within 4 days of batch culturing. In this period, ammonium-N and phosphate were reduced by more than 60 and 90 %, respectively. The high growth rate (0.57 to 1.06 day^?1) ensured a rapid removal of nutrients and thereby a short retention time. By the fourth day of cultivation, the algal biomass contained 32 % protein, but only 11 % lipids and 18 % carbohydrates. It was found that the biomass was a suitable raw material for biogas production by anaerobic digestion. Biodigestion of obtained biomass was simulated by employing the Aspen HYSYS modeling software, resulting in methane yields comparable to those found in the literature. The elemental analysis of the algal biomass showed very low concentrations of pollutants, demonstrating the potential of use of the digestate from biodigestion as a bio-fertilizer.     

Simultaneous nutrient removal and lipid production from pretreated piggery wastewater by Chlorella vulgaris YSW-04

The feasibility of using a microalga Chlorella vulgaris YSW-04 was investigated for removal of nutrients from piggery wastewater effluent. The consequent lipid production by the microalga was also identified and quantitatively determined. The wastewater effluent was diluted to different concentrations ranging from 20 to 80 % of the original using either synthetic media or distilled water. The dilution effect on both lipid production and nutrient removal was evaluated, and growth rate of C. vulgaris was also monitored. Dilution of the wastewater effluent improved microalgal growth, lipid productivity, and nutrient removal. The growth rate of C. vulgaris was increased with decreased concentration of piggery wastewater in the culture media regardless of the diluent type. Lipid production was relatively higher when using synthetic media than using distilled water for dilution of wastewater. The composition of fatty acids accumulated in microalgal biomass was dependent upon both dilution ratio and diluent type. The microalga grown on a 20 % concentration of wastewater effluent diluted with distilled water was more promising for generating high-efficient biodiesel compared to the other culture conditions. The highest removal of inorganic nutrients was also achieved at the same dilution condition. Our results revealed the optimal pretreatment condition for the biodegradation of piggery wastewater with microalgae for subsequent production of high-efficient biodiesel.     

Improving of lipid productivity of the biodiesel promising green microalga <Emphasis Type="Italic">Chlorella pyrenoidosa</Emphasis> via low-energy ion implantation

High lipid content in microalgae is an essential parameter for adopting of microalgal biomass as a feedstock for biodiesel. Mutation is one approach to obtain desired algal strain with high lipid production. In this study, a mutant strain of Chlorella pyrenoidosa was isolated using 1.5?×?10¹⁵ ions cm^?2 s^?1 of N⁺ ion beam implantation technique, which has been widely used in mutagenesis of agricultural crops. N⁺ implantation slightly improved the growth of the mutant over the corresponding wild strain with significant increase in lipid content (32.4 % higher than the wild strain), which resulted in significant increase in lipid productivity by 35 %. In addition, ion implantation mutagenesis of C. pyrenoidosa resulted in 21.4 % decrease in total saturated fatty acids (SFAs) compared to the wild type, with a noticeable increase in polyunsaturated fatty acids (PUFAs). The increase in PUFAs was due mainly to stimulation of hexadecadienoic acid (C16:2) and octadecadienoic acid (C18:2) production. However, the SFA content of wild and mutant strains was 31.7 and 24.9 % of total fatty acids, respectively, highlighting the oxidative stability of biodiesel produced by both strains according to the European standards. Cultivation of C. pyrenoidosa mutant in selenite enrichment medium for five successive cultivation experiments showed insignificant changes in biomass productivity, lipid content, and lipid productivity alongside the study period, which confirms the genetic stability of the produced mutant. The present study confirmed the feasibility of generation of microalgae mutants with significant high lipid production using ion beam implantation.     

Biodegradation of phenols by microalgae

Two green microalgae, Ankistrodesmus braunii and Scenedesmus quadricauda, degraded phenols (each tested at 400 mg ml^–1) selected from olive-oil mill wastewaters, within 5 days, with a removal greater than 70%. Green algae may, therefore, represent an alternative to other biological treatment used for the biodegradation of phenol-containing wastewaters.     

Outdoor microalgae cultivation in airlift photobioreactor at high irradiance and temperature conditions: effect of batch and fed-batch strategies,photoinhibition, and temperature stress

The microalgae Scenedesmus abundans cultivated in five identical airlift photobioreactors (PBRs) in batch and fed-batch modes at the outdoor tropical condition. The microalgae strain S. abundans was found to tolerate high temperature (35–45 °C) and high light intensity (770–1690 µmol m^− 2 s^− 1). The highest biomass productivities were 152.5–162.5 mg L^− 1 day^− 1 for fed-batch strategy. The biomass productivity was drastically reduced due to photoinhibition effect at a culture temperature of > 45 °C. The lipid compositions showed fatty acids mainly in the form of saturated and monounsaturated fatty acids (> 80%) in all PBRs with Cetane number more than 51. The fed-batch strategies efficiently produced higher biomass and lipid productivities at harsh outdoor conditions. Furthermore, the microalgae also accumulated omega-3 fatty acid (C18:3) up to 14% (w/w) of total fatty acid at given outdoor condition.

     

PHOTOBIOTREATMENT: INFLUENCE OF NITROGEN AND PHOSPHORUS RATIO IN WASTEWATER ON GROWTH KINETICS OF SCENEDESMUS OBLIQUUS

Nitrogen and phosphorus concentration in the effluent of a wastewater treatment plant can vary significantly, which could affect the growth kinetic and chemical composition of microalgae when cultivated in this medium. The aim of this work was to study the rate of growth, nutrient removal and carbon dioxide biofixation as well as biomass composition of Scenedesmus obliquus (S. obliquus) when it is cultivated in wastewater at different nitrogen and phosphorus ratio, from 1:1 to 35:1. A more homogeneous method for calculating productivities in batch reactors was proposed. The proper N:P ratio for achieving optimum batch biomass productivity ranged between 9 and 13 (263 and 322 mg L^?1 d^?1 respectively). This was also the ratio range for achieving a total N and P removal. Above and below this range (9–13) the maximum biomass concentration changed, instead of the specific growth rate.The maximum carbon dioxide biofixation rate was achieved at N:P ratio between 13 and 22 (553 and 557 mg CO2 L^?1 d^?1 respectively). Lipid and crude protein content, both depend on the aging culture, reaching the maximum lipid content (34%) at the lowest N:P (1:1) and the maximum crude protein content (34.2%) at the highest N:P (35:1).     

Microalgal system for treatment of effluent from poultry litter anaerobic digestion

The potential of mixotrophic microalgae to utilize poultry litter anaerobic digester (AD) effluent (PLDE) as nutritional growth medium was evaluated. Three algal strains viz. Chlorella minutissima, Chlorella sorokiniana and Scenedesmus bijuga and their consortium showed significant biomass productivity in 6% (v/v) concentration of PLDE in deionized water. Multiple booster dosage of PLDE supported better growth relative to a single dose PLDE. The maximum biomass productivity of 76 mg L⁻¹ d⁻¹ was recorded. The biomass was rich in protein (39% w/w) and carbohydrates (22%) while lipids (<10%) were low, making it most suitable as an animal feed supplement. The mixotrophic algae showed sustainable growth against variations in PLDE composition in different AD batches, thus proving to be a suitable candidate for large scale wastewater treatment with concomitant production of renewable biomass feedstock for animal feed and bioenergy applications.     

Biodegradation of p-nitrophenol by microalgae

A study was made on the use of a mixed microalgal consortium to degrade p-nitrophenol. The consortium was obtained from a microbial community in a waste container fed with the remains and by-products of medium culture containing substituted aromatic pollutants (nitrophenols, chlorophenols, fluorobenzene). After selective enrichment with p-nitrophenol (p-NP), followed by an antibiotic treatment, an axenic microalgal consortium was recovered, which was able to degrade p-nitrophenol. At a concentration of 50 mg L^–1, total degradation occurred within 5 days. Two species, Chlorella vulgaris var. vulgaris f. minuscula and Coenochloris pyrenoidosa, were isolated from the microalgal consortium. The species were able to accomplish p-NP biodegradation when cultured separately, although Coenochloris pyrenoidosa was more efficient, achieving the same degradation rate as the original axenic microalgal consortium. When Coenochloris pyrenoidosa was associated with Chlorella vulgaris in a 3:1 ratio, complete removal of the nitro-aromatic compound occurred within three days. This is apparently the first report on the degradation of a nitro-aromatic compound by microalgae.     

Treating Urban Wastewater: Nutrient Removal by Using Immobilized Green Algae in Batch Cultures

Essential roles of microalgae during the tertiary treatment of municipal wastewater have been proven. In order to avoid wash out of the biomass and also modify the quality of the treated wastewater; some techniques such as cell immobilization have been developed. To do so, in this study four samples from two species of microalgae (Chlorella vulgaris and Chlamydomonas sp.) were determined and confirmed by taxonomic identification. The samples were immobilized in calcium alginate beads. Within 10 days the amounts of nitrate (No₃^?-N) and orthophosphate (Po₄^3?-P) were measured to calculate the removal efficacy for each individual sample. Based on the standard methods, the amount of nitrate and orthophosphate were determined every 3 days within 10 days. The results indicate that immobilized microalgae are able to remove the nutrients and reduce the amount of nitrate and orthophosphate, significantly. Furthermore, the C. vulgaris (YG02) was the best species in this experience with 72% and 99% of reduction in the amount of nitrate and orthophosphate, respectively.     

Production and analysis of secondary carotenoids in green algae

The microalgae Neochloris wimmeri, Scenedesmus vacuolatus, Scotiellopsisoocystiformis, Chlorella zofingiensis and Protosiphon botryoides were grown under secondarycarotenoid inductive conditions. The results indicatethat nitrogen deficiency and high light intensity arepotential inducers of astaxanthin formation in thefive microalgae studied. All these microalgaeaccumulate significant quantities of secondarycarotenoids, mainly as astaxanthin esters andcanthaxanthin. They also showed high resistance toenvironmental conditions. All these qualities makethese microalgae good candidates for successfulculture in open ponds.     

Microalgae-mediated brewery wastewater treatment: effect of dilution rate on nutrient removal rates,biomass biochemical composition,and cell physiology

Microalgae have been used to remove nitrogen, phosphorus, and chemical oxygen demand (COD) from brewery wastewater (BWW). The microalga Scenedesmus obliquus was grown on BWW, using bubble column photobioreactors that operated under batch and continuous regimes. For the first time, the cell physiological status cell membrane integrity and enzymatic activity was monitored during the microalgae based BWW treatment, using flow cytometry. All the cultivations batch and continuous displayed a proportion of cells with intact membrane >?87%, although the continuous cultivations displayed a lower proportion of cells with enzymatic activity (20–40%) than the batch cultivations (97%). The dilution rate of 0.26 day^?1 was the most favorable condition, since the microalgae cultivation attained the maximum biomass productivity (0.2 g ash-free dry weight day^?1) and the total nitrogen and COD removal rates were the highest (97 and 74%, respectively), while the phosphorous removal rate was the third (23%).     

Growth of Chlorella pyrenoidosa in wastewater from cassava ethanol fermentation

In a cycle tubular photobioreactor, Chlorella pyrenoidosa was cultured in undiluted wastewater from ethanol fermentation using cassava powder as raw material. The results showed that the optimum cultivation conditions were initial pH of 6.0, temperature at 27°C, continuous illumination at 3,000 lux, and cycle speed of 110 ml min⁻¹. Under these optimum conditions, after the logarithmic phase of batch cultivation with wastewater of pH 6.0, the reactor could be continuously operated with natural pH wastewater (3.8) as feed solution. By a dilution ratio of 0.17 day⁻¹, it could be operated stably for over 30 days in continuous cultivation. pH, removal rate of chemical oxygen demand, and biomass (cell dry weight) concentration ranged from 6.22 to 6.47, 72.21 to 76.32% and 3.55 to 3.73 g l⁻¹, respectively. After treatment, the wastewater could be used again in the process of ethanol fermentation.     

Heavy metal sorption by microalgae

Viable microalgae are known to be able to accumulate heavy metals (bioaccumulation). Against a background of the increasing environmental risks caused by heavy metals, the microalgae Chlorella vulgaris and Spirulina platensis and their potential for the biological removal of heavy metals from aqueous solutions were taken as an example for investigation. Small-scale cultivation tests (50 1) with Cd-resistant cells of Chlorella vulgaris have shown that approx. 40% of the added 10 mg Cd/l was removed from the solution within seven days. At this heavy metal concentration sensitive cells died. Non-viable microalgae are able to eliminate heavy metal ions in a short time by biosorption in uncomplicated systems, without any toxicity problems. Compared with original biomasses, the sorption capacity of microalgal by-products changes only insignificantly. Their low price makes them economical.     

Nitrogen and phosphate removal by free and immobilised cells of Scenedesmus bijugatus (Kützing) from the Pinang River estuary,Penang, Malaysia

This research studied the effects of inorganic nutrient removal by free and immobilized Scenedesmus bijugatus cells, measured by algal growth (i.e., the chlorophyll a concentration) and the efficiency of the uptake of inorganic nutrients by the cells (uptake rate (b) and removal percentage) in water samples from the organically polluted Pinang River estuary (PRE). Water samples from the PRE were collected during low and high tide. S. bijugatus cells had a higher growth rate when incubated in low tide PRE water samples compared to high tide PRE water samples, with a growth rate of 0.29 µgml^?1d^?1 and 0.06 µgml^?1d^?1 for free and immobilized cells, respectively. S. bijugatus was able to more efficiently remove nitrogen, especially ammonium (81–94%), compared to phosphate (62–88%) from both low and high tide water samples. S. bijugatus cells in low tide PRE water samples recorded highest phosphate (0.36 mgL^?1d^?1 and 0.25 mgL^?1d^?1 for free and immobilized cells, respectively) and ammonium uptake rates (0.44 mgL^?1d^?1 and 0.29 mgL^?1d^?1 for free and immobilized cells respectively). Both inorganic nutrient removal and microalgal cell growth were not significantly different between free and immobilized S. bijugatus (p > 0.05). The data obtained indicated that the removal of nutrients by microalgae was affected by salinity and the immobilization technique applied may have good potential for bioremediation.