Respirometric assessment of bacterial kinetics in algae-bacteria and activated sludge processes.

Algae-bacteria (AB) consortia can be exploited for effective wastewater treatment, based on photosynthetic oxygenation to reduce energy requirements for aeration. While algal kinetics have been extensively evaluated, bacterial kinetics in AB systems are still based on parameters taken from the activated sludge models, lacking an experimental validation for AB consortia. A respirometric procedure was therefore proposed, to estimate bacterial kinetics in both activated sludge and AB, under different conditions of temperature, pH, dissolved oxygen, and substrate availability. Bacterial activities were differently influenced by operational/environmental conditions, suggesting that the adoption of typical activated sludge parameters could be inadequate for AB modelling. Indeed, respirometric results show that bacteria in AB consortia were adapted to a wider range of conditions, compared to activated sludge, confirming that a dedicated calibration of bacterial kinetics is essential for effectively modelling AB systems, and respirometry was proven to be a powerful and reliable tool to this purpose.

Productivity analysis in tubular photobioreactors using a dynamic photosynthesis model coupled to computational fluid dynamics particle tracking.

Tubular photobioreactors (TPBRs) are closed devices used for the mass culture of microalgae. TPBRs are supposed to be well-mixed, but the influence of their specific fluid dynamics in photosynthesis efficiency has never been studied in detail. Here, we use Computational Fluid Dynamics (CFD) coupled to a dynamic photosynthesis model to analyze the efficiency of the photosynthetic response in the loop of TPBRs of different sizes (14, 24, 44, 64, and 84 mm) and circulation velocities (0.4 to 1 m s-1). The results show that only the smallest diameters cause enough radial mixing for a photosynthesis-enhancing light regime (integration factor Γ = 0.199 for D = 14 mm and v = 1 m s-1) while high circulation velocities in larger diameters (up to 1 m s-1) increase operating costs but do not enhance photosynthetic productivity. It is also shown the relevance of the characteristic frequency of the strain (ß), which is crucial for high productivity.

Modelling of photosynthesis, respiration, and nutrient yield coefficients in Scenedemus almeriensis culture as a function of nitrogen and phosphorus.

Photo-respirometric tecniques are applied for evaluating photosynthetic activity in phototrophic organisms. These methods allow to evaluate photosynthetic response under different conditions. In this work, the influence of nutrient availability (nitrate, ammonium, and phosphate) on the photosynthesis and respiration of Scenedesmus almeriensis was studied using short photo-respirometric measurements. Both photosynthesis and respiration increasing until saturation value and consecutively diminishing, presenting inhibition by high concentrations. Regarding the influence of phosphorus concentration in microalgae cells, a similar hyperbolic trend was observed but no inhibition was observed at high concentration. Based on these experimental data, the respiration, and the photosynthesis rate of S. almeriensis were modelled using Haldane equation for nitrate and ammonium data, and Monod equation for phosphate data. In addition, experiments were performed to determine the yield coefficients for both nitrogen and phosphorus in S. almeriensis cultures. The data showed that the nitrogen and phosphorous coefficient yields are not constant, being modified as a function of nutrients concentration, presenting the luxury uptake phenomena. Finally, the proposed models were incorporated into a simulation tool to evaluate the photosynthetic activity and the nutrient yield coefficients of S. almeriensis when different culture media and wastewaters are used as a nitrogen and phosphorous source for its growth. Key points • Microalgal photosynthesis/respiration vary as a function of nutrients availability. • Photosynthesis inhibition appears at high N-NO 3 - and N-NH4+ concentrations. • Nutrient yield coefficients are influenced by luxury uptake phenomenon.

Analysis of productivity in raceway photobioreactor using computational fluid dynamics particle tracking coupled to a dynamic photosynthesis model.

Raceway photobioreactors (RWPs) are the most common and affordable device for the mass culture of microalgae but due to geometry and the requirement of low input power, its photosynthetic performance is low. The fluid dynamics of RWPs have been studied for information such as energy dissipation and shear rate, CFD has never been used to analyze photosynthesis efficiency by coupling dynamic photosynthesis models with microalgae trajectories. In this work, we investigate by CFD simulation the effect of circulation velocities between 0.2 and 0.8 m s-1in a 0.15 m-1 deep RWPs under standard outdoor conditions to shows that in all circumstances the RWP from the point o view of photosynthesis operates as a perfectly segregated device (no mixing) and that the average growth rate is the result of the integration of the local growth rates at different depths (integration factor Γ = 0).

Evaluation of native microalgae from Tunisia using the pulse-amplitude-modulation measurement of chlorophyll fluorescence and a performance study in semi-continuous mode for biofuel production.

BACKGROUND: Microalgae are attracting much attention as a promising feedstock for renewable energy production, while simultaneously providing environmental benefits. So far, comparison studies for microalgae selection for this purpose were mainly based on data obtained from batch cultures, where the lipid content and the growth rate were the main selection parameters. The present study evaluates the performance of native microalgae strains in semi-continuous mode, considering the suitability of the algal-derived fatty acid composition and the saponifiable lipid productivity as selection criteria for microalgal fuel production. Evaluation of the photosynthetic performance and the robustness of the selected strain under outdoor conditions was conducted to assess its capability to grow and tolerate harsh environmental growth conditions. RESULTS: In this study, five native microalgae strains from Tunisia (one freshwater and four marine strains) were isolated and evaluated as potential raw material to produce biofuel. Firstly, molecular identification of the strains was performed. Then, experiments in semi-continuous mode at different dilution rates were carried out. The local microalgae strains were characterized in terms of biomass and lipid productivity, in addition to protein content, and fatty acid profile, content and productivity. The marine strain Chlorella sp. showed, at 0.20 1/day dilution rate, lipid and biomass productivities of 35.10 mg/L day and 0.2 g/L day, respectively. Moreover, data from chlorophyll fluorescence measurements demonstrated the robustness of this strain as it tolerated extreme outdoor conditions including high (38 °C) and low (10 °C) temperature, and high irradiance (1600 µmol/m2 s). CONCLUSIONS: Selection of native microalgae allows identifying potential strains suitable for use in the production of biofuels. The selected strain Chlorella sp. demonstrated adequate performance to be scaled up to outdoor conditions. Although experiments were performed at laboratory conditions, the methodology used in this paper allows a robust evaluation of microalgae strains for potential market applications.

Evaluation of photosynthetic light integration by microalgae in a pilot-scale raceway reactor.

The improvement of photosynthetic efficiency in a 100 m2 raceway reactor by enhancement of light regime to which the cells are exposed is here reported. From Computational Fluid Dynamics it was calculated that the light exposure times ranged from 0.4 to 3.6 s while the exposure times to darkness were much longer, from 6 to 21 s. It was demonstrated that these times are too long for light integration, the cells fully adapting to local irradiances. This phenomenon was validated in the real outdoor raceway at different seasons. Simulations allows to confirm that if total light integration is achieved biomass productivity can increase up to 40 g/m2·day compared to 29 g/m2·day obtained considering local adaptation, which is close to the experimental value of 25 g/m2·day. This paper provides clear evidence of microalgae cell adaptation to local irradiance because of the unfavourable cell movement pattern in raceway reactors.

Wastewater treatment using microalgae: how realistic a contribution might it be to significant urban wastewater treatment?

Microalgae have been proposed as an option for wastewater treatment since the 1960s, but still, this technology has not been expanded to an industrial scale. In this paper, the major factors limiting the performance of these systems are analysed. The composition of the wastewater is highly relevant, and especially the presence of pollutants such as heavy metals and emerging compounds. Biological and engineering aspects are also critical and have to be improved to at least approximate the performance of conventional systems, not just in terms of capacity and efficiency but also in terms of robustness. Finally, the harvesting of the biomass and its processing into valuable products pose a challenge; yet at the same time, an opportunity exists to increase economic profitability. Land requirement is a major bottleneck that can be ameliorated by improving the system's photosynthetic efficiency. Land requirement has a significant impact on the economic balance, but the profits from the biomass produced can enhance these systems' reliability, especially in small cities.

Modeling of photosynthesis and respiration rate for Isochrysis galbana (T-Iso) and its influence on the production of this strain.

Isochrysis galbana is a widely-used strain in aquaculture in spite of its low productivity. To maximize the productivity of processes based on this microalgae strain, we have developed a model considering the influence of irradiance, temperature, pH and dissolved oxygen concentration on the photosynthesis and respiration rate. Results demonstrate that this strain tolerates temperatures up to 35 °C but it is highly sensitive to irradiances higher than 500 µE m(-2)s(-1) and dissolved oxygen concentrations higher than 11 mgl(-1). We have validated the developed model using data from an industrial-scale outdoor tubular photobioreactor demonstrating that inadequate temperature and dissolved oxygen concentrations reduce productivity to half that which is maximal, according to light availability under real outdoor conditions. The developed model is a useful tool for managing working processes, but especially in the development of new processes based on this strain and to take decisions regarding optimal control strategies.

Selection of native Tunisian microalgae for simultaneous wastewater treatment and biofuel production.

This paper focuses on the selection of native microalgae strains suitable for wastewater treatment and biofuel production. Four Chlorophyceae strains were isolated from North-eastern Tunisia. Their performances were compared in continuous mode at a 0.3 1/day dilution rate. The biomass productivity and nutrient removal capacity of each microalgae strain were studied. The most efficient strain was identified as Scenedesmus sp. and experiments at different dilution rates from 0.2 to 0.8 1/day were carried out. Maximal biomass productivity of 0.9 g/L day was obtained at 0.6 1/day. The removal of chemical oxygen demand (COD), ammonium and phosphorus was in the range of 92-94%, 61-99% and 93-99%, respectively. Carbohydrates were the major biomass fraction followed by lipids and then proteins. The saponifiable fatty acid content was in the 4.9-13.2% dry biomass range, with more than 50% of total fatty acids being composed of saturated and monosaturated fatty acids.

Utilization of secondary-treated wastewater for the production of freshwater microalgae.

In this work, we studied the potential use of secondary-treated wastewater as nutrient source in the production of freshwater microalgae strains. Experiments were performed indoors in a semicontinuous mode, at 0.3 day(-1), simulating outdoor conditions. We demonstrated that all the tested strains can be produced by using only secondary-treated wastewater as the nutrient source. The utilization of secondary-treated wastewater imposes nutrient-limiting conditions, with maximal biomass productivity dropping to 0.5 g l(-1) day(-1) and modifies the biochemical composition of the biomass by increasing the amount of lipids and carbohydrates while reducing the biomass protein content. We measured fatty acid content and productivity of up to 25 %d.wt. and 110 mg l(-1) day(-1), respectively. We demonstrated that all the tested strains were capable of completely removing the nitrogen and phosphorus contained in the secondary-treated wastewater, and while the use of this effluent reduced the cells' photosynthetic efficiency, the nitrogen and phosphorus coefficient yield increased. Muriellopsis sp. and S. subpicatus were selected as the most promising strains for outdoor production using secondary-treated wastewater as the culture medium; this was not only because of their high productivity but also their photosynthetic efficiency, of up to 2.5 %, along with nutrient coefficient yields of up to 96 gbiomass gN (-1) and 166 gbiomass gP (-1). Coupling microalgae production processes to tertiary treatment in wastewater treatment plants make it possible to recover nutrients contained in the water and to produce valuable biomass, especially where nutrient removal is required prior to wastewater discharge.

Genetic algorithm for the medium optimization of the microalga Nannochloropsis gaditana cultured to aquaculture.

A genetic algorithm has been used to optimize the composition of the culture medium for growing the microalga Nannochloropsis gaditana, based on the nutrients composition of the commercial medium ALGAL. This strategy was carried out through the implementation of 270 experiments spread over nine generations, which allowed achieving an eicosapentaenoic acid (EPA) productivity of 17.8 m gL(-1) d(-1) in a continuous culture of N. gaditana, with an increase of 23% compared to the commercial medium. The EPA yield on nitrogen and phosphorous, 0.042 and 1.146 g(EPA) g(s)(-1), respectively, were 40% and 5-fold higher, respectively, than the values obtained with the nitrogen-optimized ALGAL medium. This improvement was obtained with the medium G-8, which also allowed reducing the requirement of several nutrients such as P, Mo, Mn in 74%, 69% and 66%, respectively, as well as the thiamine content a 46%.

A quantitative study of eicosapentaenoic acid (EPA) production by Nannochloropsis gaditana for aquaculture as a function of dilution rate, temperature and average irradiance.

Different pilot-scale outdoor photobioreactors using medium recycling were operated in a greenhouse under different environmental conditions and the growth rates (0.1 to 0.5 day(-1)) obtained evaluated in order to compare them with traditional systems used in aquaculture. The annualized volumetric growth rate for Nannochloropsis gaditana was 0.26 g l(-1) day(-1) (peak 0.4 g l(-1) day(-1)) at 0.4 day(-1) in a 5-cm wide flat-panel bioreactor (FP-PBR). The biomass productivity achieved in this reactor was 10-fold higher than in traditional reactors, reaching values of 28 % and 45 % dry weight (d.w.) of lipids and proteins, respectively, with a 4.3 % (d.w.) content of eicosapentaenoic acid (EPA). A model for predicting EPA productivity from N. gaditana cultures that takes into account the existence of photolimitation and photoinhibition of growth under outdoor conditions is presented. The effect of temperature and average irradiance on EPA content is also studied. The maximum EPA productivity attained is 30 mg l(-1) day(-1).

Mixotrophic growth of Phaeodactylum tricornutum on fructose and glycerol in fed-batch and semi-continuous modes.

Mixotrophic cultures of Phaeodactylum tricornutum were carried out in bubble columns using fructose and glycerol in indoor fed-batch and semi-continuous modes. In the fed-batch cultures, different nutrient-addition strategies, combined with stepwise increments in the light intensity, were assayed. It was found that glycerol promoted significantly higher biomass productivity than fructose. A glycerol-induced photoinhibition that arrested the growth of P. tricornutun was also observed. As this was considered a limitation as regards transferring the fed-batch mode to outdoor conditions, this information was used to culture P. tricornutum in semi-continuous mode. Similar glycerol-induced photoinhibition was not observed in these cultures, even at highest dilution rates. Although the highest biomass (1.5 g L(-1) d(-1)) and EPA (40 mg L(-1) d(-1)) productivities found in the semi-continuous cultures were lower than those obtained photoautotrophically in outdoor photobioreactors, the findings showed that semi-continuous mode was an excellent candidate for transferring mixotrophic culture to an outdoor setting.

A low-cost culture medium for the production of Nannochloropsis gaditana biomass optimized for aquaculture.

Nannochloropsis gaditana is a microalga with a high nutritional value and a protein and polyunsaturated fatty acid (PUFA) content that makes it interesting as a feed in aquaculture. To maximize its productivity and nutritional value in large-scale culture, a well-known commercial medium was optimized to the most favorable nutrient level using commercial fertilizers. Optimal growth conditions were obtained in the alternative fertilizer-based medium at a nitrogen concentration of 11.3 mM, a phosphorus concentration of 0.16 mM, and a micronutrient concentration of 30 µL L(-1). This alternative medium allowed to obtain a biomass concentration similar to that achieved when using the commercial formula but with a reduction in Cu, Fe, and Mo content of 71%, 89%, and 99%, respectively. A maximum biomass productivity of 0.51 g L(-1) d(-1) was obtained. The eicosapentaenoic acid and protein contents of the biomass were 2.84% and 44% of dry weight, respectively.

Comprehensive model of microalgae photosynthesis rate as a function of culture conditions in photobioreactors.

In this paper, the influence of culture conditions (irradiance, temperature, pH, and dissolved oxygen) on the photosynthesis rate of Scenedesmus almeriensis cultures is analyzed. Short-run experiments were performed to study cell response to variations in culture conditions, which take place in changing environments such as outdoor photobioreactors. Experiments were performed by subjecting diluted samples of cells to different levels of irradiance, temperature, pH, and dissolved oxygen concentration. Results demonstrate the existence of photoinhibition phenomena at irradiances higher than 1,000 µE/m(2) s; in addition to reduced photosynthesis rates at inadequate temperatures or pH-the optimal values being 35 °C and 8, respectively. Moreover, photosynthesis rate reduction at dissolved oxygen concentrations above 20 mg/l is demonstrated. Data have been used to develop an integrated model based on considering the simultaneous influence of irradiance, temperature, pH, and dissolved oxygen. The model fits the experimental results in the range of culture conditions tested, and it was validated using data obtained by the simultaneous variation of two of the modified variables. Furthermore, the model fits experimental results obtained from an outdoor culture of S. almeriensis performed in an open raceway reactor. Results demonstrate that photosynthetic efficiency is modified as a function of culture conditions, and can be used to determine the proximity of culture conditions to optimal values. Optimal conditions found (T = 35 °C, pH = 8, dissolved oxygen concentration <20 mg/l) allows to maximize the use of light by the cells. The developed model is a powerful tool for the optimal design and management of microalgae-based processes, especially outdoors, where the cultures are subject to daily culture condition variations.

Use of secondary-treated wastewater for the production of Muriellopsis sp.

In this paper, the use of secondary-treated wastewater as the culture medium for the production of Muriellopsis sp. microalgal biomass is analyzed. Using this wastewater, a maximum biomass productivity of 0.5 g l(-1) day(-1) was measured, it being only 38 % lower than that achieved using the standard culture medium. Due to the low nitrogen content of secondary-treated wastewater, cultures produced in a medium containing a high percentage of it become nitrate-limited, thus the quantum yield reduces by up to 0.38 g E(-1)--this compares to 0.67 g E(-1) when using a standard culture medium. On the other hand, nitrate limitation enhances the accumulation of lipids and carbohydrates, with values measured at 33 and 66 % dry weight, respectively. It was also demonstrated that secondary-treated wastewater does not have any toxic effect on the growth of Muriellopsis sp. in spite of nitrogen being in the form of ammonium rather than in nitrate. Moreover, the secondary-treated wastewater was depurated when used to produce Muriellopsis sp., with the outlet biological oxygen demand and chemical oxygen demand being lower than at the inlet; the nitrate and phosphate concentrations were zero. Therefore, Muriellopsis sp. production using secondary-treated wastewater allows a reduction in the process cost by decreasing freshwater and fertilizer use, as well as by depurating the water, thus greatly enhancing process sustainability.

Medium recycling for Nannochloropsis gaditana cultures for aquaculture.

Nannochloropsis gaditana is a good producer of proteins and valuable fatty acids for aquaculture. Recycling of culture medium is interesting for microalgae commercial production as it cuts costs and prevents environmental contamination. The recycled medium must be sterilized to prevent the buildup of unwanted metabolites and microorganisms. We tested several sterilization methods: filtration, ozonation, chlorination, addition of hydrogen peroxide and heating. Results showed that the most successful method is ozonation lowering the bacterial load to 1.910(3)CFUs/mL, which is 1000-fold and 10-fold lower than the supernatant obtained after harvesting and the initial filtered medium, respectively. Continuous cultures of N. gaditana were grown using this recirculated supernatant. A maximum biomass productivity of 0.8 g/L/d composed of ∼50% proteins and 40% lipids with more than 3%d.w. EPA was obtained making this biomass very interesting for aquaculture.

Conversion of CO2 into biomass by microalgae: how realistic a contribution may it be to significant CO2 removal?

Microalgae have been proposed as a CO(2) removal option to contribute to climate change avoidance and problems coming from the use of fossil fuels. However, even though microalgae can be used to fix CO(2) from air or flue gases, they do not permit long-term CO(2) storage because they are easily decomposed. On the other hand, microalgae can contribute to an enhancement in human sustainability by producing biofuels as an alternative to fossil fuels in addition to the production of other useful chemicals and commodities. Moreover, microalgae can contribute to enhancing the sustainability of waste treatment processes, reducing the energy consumed, and improving the recycling of nutrients contained within them. This paper reviews the potential contribution of these processes and the existing knowledge in these areas.

Evaluation of flocculants for the recovery of freshwater microalgae.

The use flocculants on the recovery of freshwater microalgae is studied. Flocculants tested include metal salts, chitosan, and polyelectrolytes used in wastewater treatment processes. Influence of flocculant, but also the doses and biomass concentrations affecting biomass recovery as well as the concentration factor has been evaluated. Results showed that the use of metal salts or chitosan was not efficient, whereas polyelectrolytes allow the efficient recovery of biomass, at doses of 2-25 mg per gram of microalgae biomass. The required doses depend on the particular polyelectrolyte and the freshwater strain present; but cationic polyelectrolytes are generally recommended. The use of polyelectrolytes does not adversely affect water reuse in the production process. The concentration factors obtained are higher than 35 in most cases. Such high concentration factors allow a reduction in the equipment size necessary for biomass dewatering, thus improving the viability of using these microorganisms in biofuel or wastewater processes.

Development of a process for the production of L-amino-acids concentrates from microalgae by enzymatic hydrolysis.

A process for the production of l-amino-acids concentrates from microalgae biomass by enzymatic hydrolysis has been developed. The process includes pre-treatment for cell-disruption, enzymatic hydrolysis and final separation by centrifugation. Thermal and mechanical cell-disruption methods have been tested, selecting mechanical disruption using bead milling for 30 min. The enzymatic hydrolysis was done using the commercial enzymes Alcalase and Flavourzyme. Maximum hydrolysis was obtained for biomass concentrations under 270 g/l and previous additional treatment with Viscozyme, reaching a 42% hydrolysis. Repeated reaction steps increased the hydrolysis from 42% (4h) with a single step to 59% (8h) after two successive steps. Further increase of the number of steps had a meagre impact on the global yield. The process widens the portfolio of products that can be obtained from microalgae biomass and is a new possibility to enhance the economic viability of microalgae-based biofuels production processes.