Dual role of microalgae: Phycoremediation of domestic wastewater and biomass production for sustainable biofuels production

Global threats of fuel shortages in the near future and climate change due to green-house gas emissions are posing serious challenges and hence and it is imperative to explore means for sustainable ways of averting the consequences. The dual application of microalgae for phycoremediation and biomass production for sustainable biofuels production is a feasible option. The use of high rate algal ponds (HRAPs) for nutrient removal has been in existence for some decades though the technology has not been fully harnessed for wastewater treatment. Therefore this paper discusses current knowledge regarding wastewater treatment using HRAPs and microalgal biomass production techniques using wastewater streams. The biomass harvesting methods and lipid extraction protocols are discussed in detail. Finally the paper discusses biodiesel production via transesterification of the lipids and other biofuels such as biomethane and bioethanol which are described using the biorefinery approach.     

以市政污水为底物的微藻油脂积累和碳流分析

微藻培养的高成本阻碍了微藻生物柴油的工业化推广。文章以市政污水作为Chlorella vulgaris的培养基,分别在水力停留时间(HRT)24,48,96 h和间歇运行时对其进行培养。随着HRT的增加,C.vulgaris在系统生物群落中所占的比例增大,而且细胞密度和脂含量也分别由HRT 24 h时的0.088 g/L和8.4%提高到间歇时的0.164 g/L和14.3%。HRT对废水处理效果的影响较小,不同HRT时,COD和TP的去除率分别在66.1%~71.1%和37.6%~51.1%,TN的去除率则仅为5.9%~11.2%。对间歇污水处理过程碳元素流动分析表明,污水中的71.1%有机碳被微藻利用,其中94.6%被转化为微藻生物质,其余被代谢为CO2释放,废水中有机碳(COD)转化为微藻油脂的转化率为11.6 mg/(mg.L)。     

Culture of four microalgal strains for bioenergy production and nutrient removal in the meliorative municipal wastewater

Microalgae have been considered as the most promising sources of alternative bioenergy. For the purpose of saving costs, the present work focused on the potential use of microalgae in the meliorative municipal wastewater, which contains 90% municipal wastewater and 10% dairy wash wastewater. Four microalgal species, Palmellococcus miniatus, Neochloris oleoabundans, Scenedesmus quadricanda #507, and Chlorella zofingiensis, were cultured in pure municipal wastewater and meliorative municipal wastewater, respectively, for 5 days. Their biomass accumulation and removal rates of nitrogen and phosphate were measured. Results showed that the growth rates of Neochloris oleoabundans, Palmellococcus miniatus, and Chlorella zofingiensis in meliorative municipal wastewater (>0.8 g·L^–1·d^–1) were significantly higher than that in municipal wastewater (2.6 g·L^–1·d^–1), while there was no significant difference between the growth rates of Scenedesmus quadricanda #507 in meliorative municipal wastewater and in municipal wastewater. Neochloris oleoabundans exhibited the highest growth rate (0.86 g·L^–1·d^–1) and relatively high nutrient removal capacity. Scenedesmus quadricanda #507 had the highest P removal rate of over 94%. The four species have a similar N removal rate at about 90%. The results showed that the highest average removal rate of N and P were about 23.1 mg·L^–1·d^–1 and 7.1 mg·L^–1·d^–1. Furthermore, the content of lipid or carbohydrates increased and a different profile of fatty acids were found compared to those in municipal wastewater. Cellular components changes of microalgae in meliorative municipal wastewater were favorable as raw materials for bioethanol and biodiesel production. Cultivation with meliorative municipal wastewater is a win-win culture mode that facilitates the biomass production, lipid and carbohydrate accumulation, and wastewater purification.     

微藻生物膜去污技术应用研究进展

文章综述了微藻生物膜净化污水和生产生物燃料等方面的国内外最新成果,阐述了典型微藻去污生物膜系统的运行情况、综合效益、优缺点和推广价值,并对微藻生物膜去污技术存在的问题及关键技术进展及发展趋势进行了分析,就微藻生物膜去污技术的规模化及产业化应用提出了建议,以期为微藻生物膜去污技术的成熟和规模应用提供理论和实践支撑。     

Oxidative stress-tolerant microalgae strains are highly efficient for biofuel feedstock production on wastewater

Nutrient-rich wastewater may provide a sustainable means to cultivate microalgal biomass for biofuel use, yet many microalgal strains are very sensitive to wastewater due to toxicity caused by abiotic and biotic stresses. Naturally adapted strains that can efficiently grow in wastewater effluent are therefore of interest, however, the mechanisms by which such strains tolerate wastewater conditions are unknown. This study isolated indigenous chlorophyte microalgae strains from a municipal secondary wastewater effluent tank. The strains were identified by molecular phylogenetics and characterised by their ability to utilise exogenous organic carbon sources for mixotrophic growth and on the basis of oxidative stress tolerance, in order to elucidate the mechanisms of wastewater adaptation. Two of the strains, identified as Chlorella luteoviridis and Parachlorella hussii, could grow very well in raw wastewater due to their substantial tolerance to oxidative stress, which is highly induced by the wastewater environment. These strains exhibited high ascorbate peroxidase activity allowing increased scavenging of reactive oxygen species compared to strains that are not well adapted to the wastewater conditions. Both strains displayed high biomass and lipid productivity values in wastewater effluent. The accumulated lipids were suitable for biodiesel usage with characteristics equivalent to palm oil- and sunflower oil-derived biodiesel. The strains were also efficient in nutrient remediation from the wastewater. These results demonstrate the potential of these two strains for future biofuel applications coupled to wastewater remediation and highlight the importance of oxidative stress tolerance as a key indicator of efficient wastewater growth.     

Biohydrogen production by a novel integration of dark fermentation and mixotrophic microalgae cultivation

Biohydrogen is usually produced via dark fermentation, which generates CO₂ emissions and produces soluble metabolites (e.g., volatile fatty acids) with high chemical oxygen demand (COD) as the by-products, which require further treatments. In this study, mixotrophic culture of an isolated microalga (Chlorella vulgaris ESP6) was utilized to simultaneously consume CO₂ and COD by-products from dark fermentation, converting them to valuable microalgae biomass. Light intensity and food to microorganism (F/M) ratio were adjusted to 150 μmol m⁻² s⁻¹ and F/M ratio, 4.5, respectively, to improve the efficiency of assimilating the soluble metabolites. The mixotrophic microalgae culture could reduce the CO₂ content of dark fermentation effluent from 34% to 5% with nearly 100% consumption of soluble metabolites (mainly butyrate and acetate) in 9 days. The obtained microalgal biomass was hydrolyzed with 1.5% HCl and subsequently used as the substrate for bioH₂ production with Clostridium butyricum CGS5, giving a cumulative H₂ production of 1276 ml/L, a H₂ production rate of 240 ml/L/h, and a H₂ yield of 0.94 mol/mol sugar.     

Biomass and lipid production of marine microalgae using municipal wastewater and high concentration of CO2

In order to reduce the cost of the production of microalgae for biodiesel, the feasibility of using the mixture of seawater and municipal wastewater as culture medium and CO₂ from flue gas for the cultivation of marine microalgae was investigated in this study. Effects of different ratios of municipal wastewater and 15% CO₂ aeration on the growth of Nannochloropsis sp. were examined, and lipid accumulation of microalgae was also studied under nitrogen starvation and high light. It was found that optimal growth of microalgae occurred in 50% municipal wastewater, and the growth was further significantly enhanced by aeration with 15% CO₂. When Nannochloropsis sp. cells were transferred from the first growth phase to the second lipid accumulation phase under the combination of nitrogen deprivation and high light, both biomass and lipid production of Nannochloropsis sp. were significantly increased. After 12 days of the second-phase cultivation, the biomass concentration and total lipid content increased from 0.71 to 2.23 g L⁻¹ and 33.8–59.9%, respectively. This study suggests that it is possible to utilize municipal wastewater to replace nutrients in seawater medium and use flue gas to provide CO₂ in the cultivation of oil-bearing marine microalgae for biodiesel.     

The effects of ultraviolet radiation on growth,biomass, lipid accumulation and biodiesel properties of microalgae

The effect of UV light on growth, biomass, lipid accumulation and biodiesel properties of microalgae was studied. A Microalgae strain Chlorella sorokiniana UUIND6 was cultivated for 14 days as under LED light (Control) and microalgae were exposed to UV light (280–320 nm) in the middle of the photoperiod for 3 days. The growth rate of microalgae was analyzed by spectrophotometer and cell counting, while oil accumulation was analyzed by improved Nile red method. Results showed that microalgae under UV light treated algal cells showed less growth. FAMEs profile of UV treated algal cells mainly contains hexadecanoic acid (C16), stearic acid (C18) fatty acids. PUFA found in very less amount in UV treated cells as compared to control.     

Application of experimental design methodology for optimization of biofuel production from microalgae

Optimization of biofuel productivity, in terms of lipid content, polysaccharide content, and calorific value, from microalgae was performed by varying four variables (temperature, light intensity, nitrogen content, and CO₂ addition) using a 2⁴ full factorial design. A statistical analysis showing the influence of each variable and their interactions was conducted. The selected variables all influence biofuel productivity, but their importance varies according to the sequence: CO₂ addition > temperature > nitrogen content > light intensity. Interactive effects of temperature with light intensity and nitrogen with CO₂ addition for lipid and polysaccharide productivities were identified, respectively. For calorific value, interactive effects of CO₂ addition with light intensity and nitrogen content were observed. The highest biofuel productivity was obtained at the following conditions: temperature (>25 °C), light intensity (>60 μmol photons m⁻² s⁻¹), nitrogen content (<50 mg L⁻¹), and CO₂ addition (>18 mL L⁻¹ d⁻¹). 10 days was found to be the most favorable cultivation time for lipid production under the investigated conditions.     

Comparison study of thermochemical waste-heat recuperation by steam reforming of liquid biofuels

The concept of thermochemical exhaust heat recuperation by steam reforming of biofuels is considered. Thermochemical recuperation can be considered as an on-board hydrogen production technology. A schematic diagram of a fuel-consuming equipment with thermochemical heat recuperation is described. The thermodynamic analysis of the thermochemical recuperation systems was performed to determine the efficiency of using various fuels, in particular, methanol, ethanol, n-butanol, and glycerol. The thermodynamic analysis was performed by Gibbs free energy minimization method and implemented using the Aspen Hysys program. The thermodynamic analysis was performed for a wide temperature range from 400 to 900 K, for steam-to-fuel of 1, and pressures of 1 bar. The maximum fuel conversion reaches for the following temperatures: methanol - 600 K, ethanol - 730 K, n-butanol - 860 K, glycerol - 890 K. The dependence of the reforming enthalpy on temperature is determined. It was shown that the reaction enthalpy determines the heat transformation coefficient, which shows the ratio of the low heat value of synthetic fuel and the low heat value of the initial fuel. For all studied fuels, the maximum value of the transformation coefficient is observed for steam reforming of ethanol and the maximum heat transformation coefficient is 1.187. The temperature range is determined at which the maximum efficiency of the use of thermochemical recuperation occurs due to the reforming of biofuels. For methanol, the effective temperature is about 600 K, for ethanol is about 700 K, for n-butanol is 850 K, for glycerol is more than 900 K. The results obtained make it possible to efficiently select the type of fuel for thermochemical recuperation due to steam reforming.     

Biohydrogen production from pretreated sludge and synthetic and real biodiesel wastewater by dark fermentation

Different types of sludge pretreatments were tested, with thermal shock at 90°C to 95°C for 60 minutes plus a 6‐hour rest period achieving the best results for inhibition of methanogen microorganisms and inoculum enrichment with H₂‐producing bacteria, which produced a H₂‐rich biogas (up to 65% mol/mol) without the presence of CH₄. Wastewater from biodiesel production (WBP), containing mainly methanol (128 g/L) and glycerol (4 g/L), was evaluated as a potential substrate to produce H₂ through dark fermentation. Both methanol‐based solutions and methanol‐rich wastewater were not suitable for hydrogen production; however, these effluents showed a strong potential for CH₄‐rich biogas production. A fractional factorial design was employed to evaluate the effect of six substrate‐related variables (glycerol content, 25% and 75%; COD content, 4 and 50 g/L, COD:VSS ratio, 1:1 and 5:1; COD:N:P ratio, 350:0:0 and 350:5:1; NaCl content, 0.5 and 12.0 g/L; and pH, 4.0 and 5.5) on the H₂ production from glycerol‐methanol–based synthetic solutions (synthetic WBP). Some substrate‐related variables had a crucial impact on the hydrogen production potential from WBP, which was significantly affected by the COD and salinity content in the substrate. WBP containing high glycerol (representing until 75% of the COD) and salinity (up to 12 g/L as NaCl) content could be turned into a potential substrate for H₂ production through dark fermentation as long as specific fermentation conditions are maintained, such as pH 5.5 to 5.7 and a substrate COD content up to 50 g/L. Using this condition, glycerol conversion, H₂ productivity, and H₂ yield of 81.3 ± 8.9%, 102.8 ± 18.2 mL H₂/L.d, and 24.5 ± 4.4 mL H₂/g COD_applied, respectively, were obtained.     

Optimization of conditions for hydrogen production from complex dairy wastewater by anaerobic sludge using desirability function approach

Present study deals with the multiple-response optimization for biohydrogen production using anaerobic sludge and outstanding approach to overcome the drawbacks of conventional response surface methodology (RSM). Dairy wastewater was used as source in batch fermentation was followed for this study. Response surface methodology (RSM), based on a three level, four variable Box–Behnken design, was employed to obtain the best possible combination of substrate concentration, pH, COD/N ratio and COD/P ratio for maximum H₂ yield (HY) and specific hydrogen production rate (SHPR). Experimental data were evaluated by applying RSM integrating a desirability function approach. The optimum H₂ yield and SHPR conditions were: substrate concentration 15.3 g COD/L, pH 5.5, COD/N ratio 100.5 and COD/P ratio 120 with maximum overall desirability D of 0.94. The confirmation experiment under these optimal condition showed a HY and SHPR of 13.54 mmol H₂/g COD and 29.91 mmol H₂/g-VSS.d, respectively. This was only 0.22% and 0.20%, respectively, different from the predicted values, suggesting that the desirability function approach with RSM was a useful technique to get the maximum H₂ yield and SHPR simultaneously.     

A comprehensive review on pretreatment of microalgae for biogas production

Use of microalgal biomass for renewable energy production has gained considerable attention in the world due to increasing global energy demand and negative environmental impacts of nonrenewable fossil fuels. Anaerobic digestion is one of the renewable technologies that microalgal biomass is converted into biogas by anaerobic archea. One of the main drawbacks of using microalgal biomass for biogas production is that certain types of microalgae has rigid cell wall characteristics, which limits accessibility of anaerobic archea to microalgal intracellular organic matter during hydrolysis phase. This limitation lowers efficiency of biogas production from microalgal biomass. However, introducing pretreatment methods prior to anaerobic digestion provides disruption of rigid microalgal cell wall and improve biogas yields from microalgal biomass. The objective of this paper was to review current knowledge related to pretreatment methods applied prior to anaerobic digestion of microalgal biomass. Efficiency and applicability of pretreatment methods mainly depend on type of microalgae, cell wall characteristics, and cost and energy requirements during pretreatment process. In this review, various type of pretreatment methods applied to microalgal biomass was discussed in detail with background knowledge and literature studies in their potential on maximization of biogas yields and their cost effectiveness, which is important for large‐scale applications. In the view of current knowledge, it was concluded that each pretreatment method has a relative contribution to improvement in biogas production depending on the type of microalgae. However, energy and cost requirements are the main limitations for pretreatment. So, further studies should focus on reduction of cost and energy demand by introducing combined methods, novel chemicals, and on‐site or immobilized enzymes in pretreatment to increase feasibility of pretreatment prior to anaerobic digestion in industrial scale.     

Comparison of open-air and semi-enclosed cultivation system for massive microalgae production in sub-tropical and temperate latitudes

This study compared open-air and semi-enclosed production system of the marine microalgae Nannochloropsis oculata in a sub-tropical region (32°S; 52°W) under uncontrolled environmental conditions. The semi-enclosed system was composed of 1.2 m³ circular tanks installed inside of a greenhouse. Water temperature was 4 °C higher in the indoor treatment than in the outdoor, mainly in winter although no difference was observed in warmer seasons. Moreover, variation in salinity was observed in the outdoor treatment due to rainfall (winter) and evaporation (spring), whereas indoor treatment experienced an increase (up to 100 PSU) due to evaporation only in warmer seasons. Light transmission was approximately 20% lower in the indoor treatment although cell densities and biomass yields were higher indoor during winter. As the temperature increased (spring) no differences were observed among treatments. In summary, partial control of temperature and salinity in the semi-enclosed system, especially during the colder and rainy season, allowed higher microalgae biomass production. Further experiments must be conducted with CO₂ addition, larger pH range and salinity control.     

Hydrogen gas production from vinegar fermentation wastewater by electro-hydrolysis: Effects of initial COD content

Vinegar fermentation wastewater with different initial COD contents (9.66–48.6 g L⁻¹) were used for hydrogen gas production with simultaneous COD removal by electro-hydrolysis. The applied DC voltage was constant at 4 V. The highest cumulative hydrogen production (3197 ml), hydrogen yield (2766 ml H₂ g⁻¹ COD), hydrogen formation rate (799 ml d⁻¹), and percent hydrogen (99.5%) in the gas phase were obtained with the highest initial COD of 48.6 g COD L⁻¹. The highest energy efficiency (48%) was obtained with the lowest COD content of 9.66 g L⁻¹. Hydrogen gas production by water electrolysis was less than 250 ml and wastewater control resulted in less than 25 ml H₂ in 96 h. The highest (12%) percent COD removal was obtained with the lowest COD content. Hydrogen gas was produced by reaction of (H⁺) ions present in raw WW ( pH = 3.0) and protons released from acetic acid with electrons provided by electrical current. Electro-hydrolysis of vinegar wastewater was proven to be an effective method of H₂ gas production with some COD removal.     

Prospects of using microalgae for biofuels production: Results of a Delphi study

Advanced biofuels, such as those obtained from microalgae, are widely accepted as better choices for achieving goals of incorporating renewables and non-food fuel sources into the transportation sector, and for overcoming land use issues due to biofuel crops. Main challenges are currently the feasibility of large-scale commercialization of microalgae biofuels, since there are still some technical problems to overcome (e.g. the high energy consumption associated with biomass processing) and the majority of economic and financial analyses are based on pilot-scale projects. Therefore, this article presents the results of a Delphi study aiming to identify the main obstacles and most critical issues affecting the potential of large-scale commercialization of microalgae biodiesel and its incorporation into the fuel market. According to the authors' knowledge, this is the first Delphi study with this objective. The respondents are worldwide market specialists in the survey themes that ranged from biofuels economics to their environmental sustainability. One of the key findings is that most of the experts believe that production of microalgae biofuels will achieve its full commercial scale until 2020, and that from 2021 till 2030 it could represent from 1% to 5% of the worldwide fuel consumption. The study results also showed that environmental issues are where expert opinion differs more.     

Effective extraction of microalgae lipids from wet biomass for biodiesel production

Producing biodiesel from lipid extracted from microalgae is a promising approach for sustainable fuel production. However, this approach is not yet commercialized due to the high costs of upstream processes that are associated with the time consuming and/or energy intensive drying, and lipid extraction processes. In this study, the possibility of avoiding the drying process, and extracting the lipid directly from the wet concentrated cells, using enzymatic disruption to enhance the extraction, has been tested. Results showed that lysozyme and cellulase were both efficient in disrupting cell walls and enhancing lipid extraction from wet samples, with highest lipid extraction yield of 16.6% achieved using lysozyme. The applicability of using supercritical CO₂ (SC-CO₂) in extracting lipid from wet biomass was also tested and the highest yield of 12.5% was achieved using lysozyme. In addition, a two-step culturing process was applied, using Scenedesmus sp., to combine both high biomass growth and lipid content. The strain was able to increase its biomass productivity in the first stage, reaching 174 mg l⁻¹ d⁻¹, with almost constant lipid content. In the second stage, the lipid content was enhanced by six-fold after three weeks of nitrogen starvation, but with lower biomass productivity.     

Hydrogen production from the tannery wastewater treatment by using agriculture supports membrane/adsorbents electrochemical system

The tannery is one of the oldest and most popular industries in the world. It is also characterized as pollutants generated industries which discharge toxic chemical output effluents to the environment. The process of tannery included a wide variety of chemical and inorganic constituents. This work focused on the removal of Chromium heavy metals as well as producing clean energy from the treatment of tannery wastewater. Accordingly, the analysis result on the input effluents appeared to have a brown color with a very high COD value, a high concentration of Chromium heavy metals as well as other organic compounds. After collected from the source, the effluents were settling and applied a simple subsequent filtration with lab scale cloth filter, the filtered effluents then were treated in an electrochemical system. Throughout many experiments, we introduce an electrochemical system with 5 × 5 cm electrodes (Platinum coated panel anode and carbon fiber cloth cathode), the low input voltage (10 V), easy setup and the support of separator membrane/adsorbents placed in the middle of the system to enhance the removal rates of heavy metals.It was found that the performance of the electrochemical system is under the influence of various factors such as temperature, pH value, adsorbents dose, and apply voltage. During 48 h of treatment, almost 80% of Chromium metals were treated by means of adsorption and electrical reduction process. The generation rate of hydrogen gases during the electrolysis process was also notable (45–65 cc/min). Furthermore, the adsorbent materials were still intact and seem to be ready for a longer run. The study also observed the adsorbent membrane of bagasse and straw showed their best removal efficiencies over other candidates. In that manner, we successfully provide an effective method for heavy metals removal and also capable of generating clean energy. The analysis results clarify that most of the parameters of the physical and chemical result were found well below the prescribed permissible limits of discharged effluents follows the standard of industrial waste management in Vietnam.     

Kinetic analysis of biohydrogen production from complex dairy wastewater under optimized condition

Present work describes a kinetic analysis of various aspects of biohydrogen production in batch test using optimized conditions obtained previously. Monod model and Logistic equation have been used to find growth kinetic parameters in batch test under uncontrolled pH. The values of μ_m, K_s, and X_m were 0.64 h⁻¹, 15.89 g-COD L⁻¹, and 7.26 g-VSS L⁻¹, respectively. Modified Leudeking-Piret and Michaelis–Menten equation corroborates a flux of energy to hydrogen production pathway and energy sufficiency in the system. Modified Gompertz equation illustrates that the overall rate and hydrogen yield at 15 g-COD L⁻¹ was higher compared to a dark fermentation of other wastewaters. Besides, Andrew's equation also suggests that since the higher value of K_I (19.95 g-COD L⁻¹), k (255 mL h⁻¹ L⁻¹) was not inhibited at high S. The experimental results implied that the entire products during the fermentation process were growth and substrate degradation associated. The result also confirms that the acetate and butyrate were substantially used for hydrogen production in acidogenic metabolism under uncontrolled pH.     

Microalgae-microbial fuel cell (mMFC): an integrated process for electricity generation,wastewater treatment,CO2 sequestration and biomass production

The world today is facing a crisis of energy and environmental pollution. Conventional or photosynthetic microbial fuel cell (MFC) is an advanced “green” energy technology that utilizes living microorganisms to convert biochemical or light energy into electricity through metabolic reaction and photosynthesis, offering a potential solution for the above-mentioned crisis. Further incorporating microalgae into MFC, microalgae-microbial fuel cell (mMFC) integrates electricity generation, wastewater treatment, CO₂ sequestration and biomass production in a single, self-sustainable technology. This review first describes the fundamentals of MFC as well as its applications in treating domestic, municipal, agricultural and industrial wastewaters. Then, mMFC-based configurations and applications with its advantages compared with MFC are explained in particular, together with the parameters governing its performance. Lastly, the opportunities and challenges involved in the development of mMFCs are also explored.