Effects of Temperature, Light and Salt on the Production of Fucoxanthin from Conticribra weissflogii.

Fucoxanthin is a natural active substance derived from diatoms that is beneficial to the growth and immunity of humans and aquatic animals. Temperature, light and salinity are important environmental factors affecting the accumulation of diatom actives; however, their effects on the production of fucoxanthin in C. weissflogii are unclear. In this study, single-factor experiments are designed and followed by an orthogonal experiment to determine the optimal combination of fucoxanthin production conditions in C. weissflogii. The results showed that the optimum conditions for fucoxanthin production were a temperature of 30 °C, a light intensity of 30 umol m-2 s-1 and a salinity of 25. Under these conditions, the cell density, biomass, carotenoid content and fucoxanthin content of C. weissflogii reached 1.97 × 106 cell mL-1, 0.76 g L-1, 2.209 mg L-1 and 1.372 mg g-1, respectively, which were increased to 1.53, 1.71, 2.50 and 1.48 times higher than their initial content. The work sought to give useful information that will lead to an improved understanding of the effective method of cultivation of C. weissflogii for natural fucoxanthin production.

The contribution of bacteria to algal growth by carbon cycling.

Algal mass production in open systems is often limited by the availability of inorganic carbon substrate. In this paper, we evaluate how bacterial driven carbon cycling mitigates carbon limitation in open algal culture systems. The contribution of bacteria to carbon cycling was determined by quantifying algae growth with and without supplementation of bacteria. It was found that adding heterotrophic bacteria to an open algal culture dramatically enhanced algae productivity. Increases in algal productivity due to supplementation of bacteria of 4.8 and 3.4 times were observed in two batch tests operating at two different pH values over 7 days. A kinetic model is proposed which describes carbon limited algal growth, and how the limitation could be overcome by bacterial activity to re-mineralize photosynthetic end products.

From lab to application: Cultivating limnetic microalgae in seawater coupled with wastewater for biodiesel production on a pilot scale.

The technology of cultivating salt-tolerant limnetic microalgae in seawater reduces the freshwater demand and costs of biodiesel production. However, all current trials still occur on the bench scale, and efforts for pilot-scale operation are urgently needed. This study firstly optimised the diameter of the photobioreactors (PBRs) to 0.2 m, as the single module to produce lipid-rich Golenkinia sp. SDEC-16 because of the better algal growth and light attenuation in the PBRs, and then established a 1000 L algal cultivation system. In a medium of seawater supplemented with monosodium glutamate wastewater at a ratio of 1:1000 (S-MSGW), the biomass productivity was 0.26 g/L/d, which was approaching the 0.30 g/L/d obtained in BG11, and the lipid productivity (98.99 mg/L/d) was doubled in comparison to growth in BG11. C16-C18 accounted for more than 98% of the total fatty acid in S-MSGW, and the biodiesel properties also met the biodiesel standards. The input cost of the biodiesel in this pilot-scale system was estimated to be 2.2 $/kg. When considering the carbon reduction and diversified application of the biomass, Golenkinia sp. would annually capture 186.77 kg/m3 PBR of CO2, and yield an output-to-input ratio (OIR) of 3.4 in S-MSGW, higher than the 2.8 in BG11.

Lipid production characteristics of a newly isolated microalga Asterarcys quadricellulare R-56 as biodiesel feedstock.

In this study, a new microalgal strain, Asterarcys quadricellulare R-56, was isolated for biomass and lipid production. The effects of carbon and nitrogen sources and initial pH on the cell growth and lipid accumulation of strain R-56 were investigated. At 10 g L-1 glucose, 0.6 g L-1 sodium nitrate, and pH 7, the highest biomass of 4.18 g L-1 and lipid content of 43.66% were obtained. Microalgae had a broad pH tolerance in the range of 5-11, and the pH of the culture medium was close to neutral at the end of cultivation. The maximum contents of chlorophyll, carbohydrate, and protein under the recommended culture conditions were 19.47 mg mL-1, 21.80%, and 29.94%, respectively. Palmitic and palmitoleic acid contents in strain R-56 accounted for as high as 83.73% of total fatty acids. This study suggested that strain R-56 was a promising lipid producer for high-quality biodiesel production.

Optimal influent N-to-P ratio for stable microalgal cultivation in water treatment and nutrient recovery.

Species specific nitrogen-to-phosphorus molar ratio (NPR) has been suggested for green microalgae. Algae can store nitrogen and phosphorus, suggesting that the optimum feed concentration dynamically changes as function of the nutrient storage. We assessed the effect of varying influent NPR on microalgal cultivation in terms of microbial community stability, effluent quality and biokinetics. Mixed green microalgae (Chlorella sorokiniana and Scenedesmus sp.) and a monoculture of Chlorella sp. were cultivated in continuous laboratory-scale reactors treating used water. An innovative image analysis tool, developed in this study, was used to track microbial community changes. Diatoms proliferated as influent NPR decreased, and were outcompeted once cultivation conditions were restored to the optimal NPR range. Low NPR operation resulted in decrease in phosphorus removal, biomass concentration and effluent nitrogen concentration. ASM-A kinetic model simulation results agreed well with operational data in the absence of diatoms. The failure to predict operational data in the presence of diatoms suggest differences in microbial activity that can significantly influence nutrient recovery in photobioreactors (PBR). No contamination occurred during Chlorella sp. monoculture cultivation with varying NPRs. Low NPR operation resulted in decrease in biomass concentration, effluent nitrogen concentration and nitrogen quota. The ASM-A model was calibrated for the monoculture and the simulations could predict the experimental data in continuous operation using a single parameter subset, suggesting stable biokinetics under the different NPR conditions. Results show that controlling the influent NPR is effective to maintain the algal community composition in PBR, thereby ensuring effective nutrients uptake.

Integration of Algae to Improve Nitrogenous Waste Management in Recirculating Aquaculture Systems: A Review.

This review investigates the performance and the feasibility of the integration of an algal reactor in recirculating aquaculture systems (RAS). The number of studies related to this topic is limited, despite the apparent benefit of algae that can assimilate part of the inorganic waste in RAS. We identified two major challenges related to algal integration in RAS: first, the practical feasibility for improving nitrogen removal performance by algae in RAS; second, the economic feasibility of integrating an algal reactor in RAS. The main factors that determine high algal nitrogen removal rates are light and hydraulic retention time (HRT). Besides these factors, nitrogen-loading rates and RAS configuration could be important to ensure algal performance in nitrogen removal. Since nitrogen removal rate by algae is determined by HRT, this will affect the size (area or volume) of the algal reactor due to the time required for nutrient uptake by algae and large surface area needed to capture enough light. Constraints related to design, space, light capture, and reactor management could incur additional cost for aquaculture production. However, the increased purification of RAS wastewater could reduce the cost of water discharge in places where this is subject to levees. We believe that an improved understanding of how to manage the algal reactor and technological advancement of culturing algae, such as improved algal reactor design and low-cost artificial light, will increase the practical and economic feasibility of algal integration in RAS, thus improving the potential of mass cultivation of algae in RAS.

Enhanced simultaneous removal of nitrogen, phosphorous, hardness, and methylisothiazolinone from reverse osmosis concentrate by suspended-solid phase cultivation of Scenedesmus sp. LX1.

The disposal of reverse osmosis (RO) concentrate (ROC) is a critical challenge impeding the application of RO-based wastewater reclamation. Herein, we proposed an enhanced biotreatment approach for the simultaneous removal of nitrogen, phosphorous, hardness, and methylisothiazolinone (MIT) from ROC by suspended-solid phase cultivation of Scenedesmus sp. LX1. Repeated carrier addition, guided by the developed optimal carrier addition model, efficiently enhanced algal growth and contaminant removal through dynamically controlling the suspended algal density by cell attachment. The maximum algal growth rate (212.2 mg/(L∙d)) increased by 41% compared with the control, and the time needed for reaching the maximum algal biomass (906.7 mg/L) was shortened by 1 d, attributing to the mitigation of density restriction. 91.8% of nitrogen (30.2 mg/L) was removed with 5.5 mg/(L∙d) accelerating removal rate, and phosphate (3.7 mg/L) was completely removed within 1 d. Hardness precursors calcium and inorganic carbon were also removed in large amounts, 268.4 and 128.2 mg/L, respectively. Moreover, suspended-solid phase cultivation significantly mitigated the growth inhibition caused by MIT toxicity, enabled the algae to completely biodegrade MIT of extremely high concentrations (4.7 mg/L and 11.4 mg/L) in a short time. Our results demonstrate the feasibility of suspended-solid phase algal cultivation for simultaneously and effectively removing multiple main contaminants from ROC.

A Phosphite Dehydrogenase Variant with Promiscuous Access to Nicotinamide Cofactor Pools Sustains Fast Phosphite-Dependent Growth of Transplastomic Chlamydomonas reinhardtii.

Heterologous expression of the NAD+-dependent phosphite dehydrogenase (PTXD) bacterial enzyme from Pseudomonas stutzerii enables selective growth of transgenic organisms by using phosphite as sole phosphorous source. Combining phosphite fertilization with nuclear expression of the ptxD transgene was shown to be an alternative to herbicides in controlling weeds and contamination of algal cultures. Chloroplast expression of ptxD in Chlamydomonas reinhardtii was proposed as an environmentally friendly alternative to antibiotic resistance genes for plastid transformation. However, PTXD activity in the chloroplast is low, possibly due to the low NAD+/NADP+ ratio, limiting the efficiency of phosphite assimilation. We addressed the intrinsic constraints of the PTXD activity in the chloroplast and improved its catalytic efficiency in vivo via rational mutagenesis of key residues involved in cofactor binding. Transplastomic lines carrying a mutagenized PTXD version promiscuously used NADP+ and NAD+ for converting phosphite into phosphate and grew faster compared to those expressing the wild type protein. The modified PTXD enzyme also enabled faster and reproducible selection of transplastomic colonies by directly plating on phosphite-containing medium. These results allow using phosphite as selective agent for chloroplast transformation and for controlling biological contaminants when expressing heterologous proteins in algal chloroplasts, without compromising on culture performance.

Cogeneration of hydrogen and lipid from stimulated food waste in an integrated dark fermentative and microalgal bioreactor.

In this study, a novel integrated dark fermentative and microalgal bioreactor (IDFMB) was developed to simultaneously produce H2 and lipid from food waste. Under the optimized working volume ratio of 1:4, starch concentration of 7 g L-1 and initial pH of 7.0, the highest H2 production of 1643.5 mL L-1 and lipid yield of 515.6 mg L-1 were achieved. Microalgae can effectively utilize the main end products in dark fermentative effluent (acetic acid and butyric acid) for cell growth and lipid accumulation. Compared with single dark fermentation, the energy conversion efficiency from stimulated food waste was significantly enhanced by the IDFMB, which increased from 14.8% to 35%. Microbial community analysis revealed that Clostridium was the dominant bacteria for H2 generation, and the IDFMB can improve the survival environment of microorganisms. This study provides a novel strategy for efficient energy recovery from food waste.

Continuous energy recovery and nutrients removal from molasses wastewater by synergistic system of dark fermentation and algal culture under various fermentation types.

Synergistic system of dark fermentation and algal culture was initially operated at batch mode to investigate the energy production and nutrients removal from molasses wastewater in butyrate-type, ethanol-type and propionate-type fermentations. Butyrate-type fermentation was the most appropriate fermentation type for the synergistic system and exhibited the accumulative hydrogen volume of 658.3 mL L-1 and hydrogen yield of 131.7 mL g-1 COD. By-products from dark fermentation (mainly acetate and butyrate) were further used to cultivate oleaginous microalgae. The maximum algal biomass and lipid content reached 1.01 g L-1 and 38.5%, respectively. In continuous operation, the synergistic system was stable and efficient, and energy production increased from 8.77 kJ L-1 d-1 (dark fermentation) to 17.3 kJ L-1 d-1 (synergistic system). Total COD, TN and TP removal efficiencies in the synergistic system reached 91.1%, 89.1% and 85.7%, respectively. This study shows the potential of the synergistic system in energy recovery and wastewater treatment.

Maximization of cell growth and lipid production of freshwater microalga Chlorella vulgaris by enrichment technique for biodiesel production.

Chlorella vulgaris was cultivated under limitation and starvation and under controlled conditions using different concentrations of nitrate (NaNO3) and phosphate (K2HPO4 and KH2PO4) chemicals in modified Bold basal medium (BBM). The biomass and lipid production responses to different media were examined in terms of optical density, cell density, dry biomass, and lipid productivity. In the 12-day batch culture period, the highest biomass productivity obtained was 72.083 mg L-1 day-1 under BBM - NcontrolPlimited condition. The highest lipid content, lipid concentration, and lipid productivity obtained were 53.202 %, 287.291 mg/L, and 23.449 mg L-1 day-1 under BBM - NControlPDeprivation condition, respectively. Nitrogen had a major effect in the biomass concentration of C. vulgaris, while no significant effect was found for phosphorus. Nitrogen and phosphorus starvation was found to be the strategy affecting the lipid accumulation and affected the lipid composition of C. vulgaris cultures.

Harvesting of microalgal biomass: Efficient method for flocculation through pH modulation.

Harvesting of the micro alga Chlorococcum sp. R-AP13 through autoflocculation, chemical flocculants or by change in medium pH was evaluated. Surface charge of algal cells changed in response to the method used and affected flocculation efficiency. While aluminum sulfate and FeCl3 supported 87% and 92% efficiency, auto flocculation could recover 75% of biomass in 10min. Maximum efficiency (94%) was obtained with change in medium pH from 8.5 to 12.0 achieved through addition of 40mgl(-1) of NaOH. Since high concentrations of FeCl3 and AlSO4 were toxic to the cells, flocculation induced by pH change may be considered the most effective strategy. Residual medium after flocculation could be reused efficiently for algal cultivation, minimizing the demand for fresh water.

Life cycle assessment as development and decision support tool for wastewater resource recovery technology.

Life cycle assessment (LCA) has been increasingly used in the field of wastewater treatment where the focus has been to identify environmental trade-offs of current technologies. In a novel approach, we use LCA to support early stage research and development of a biochemical system for wastewater resource recovery. The freshwater and nutrient content of wastewater are recognized as potential valuable resources that can be recovered for beneficial reuse. Both recovery and reuse are intended to address existing environmental concerns, for example, water scarcity and use of non-renewable phosphorus. However, the resource recovery may come at the cost of unintended environmental impacts. One promising recovery system, referred to as TRENS, consists of an enhanced biological phosphorus removal and recovery system (EBP2R) connected to a photobioreactor. Based on a simulation of a full-scale nutrient and water recovery system in its potential operating environment, we assess the potential environmental impacts of such a system using the EASETECH model. In the simulation, recovered water and nutrients are used in scenarios of agricultural irrigation-fertilization and aquifer recharge. In these scenarios, TRENS reduces global warming up to 15% and marine eutrophication impacts up to 9% compared to conventional treatment. This is due to the recovery and reuse of nutrient resources, primarily nitrogen. The key environmental concerns obtained through the LCA are linked to increased human toxicity impacts from the chosen end use of wastewater recovery products. The toxicity impacts are from both heavy metals release associated with land application of recovered nutrients and production of AlCl3, which is required for advanced wastewater treatment prior to aquifer recharge. Perturbation analysis of the LCA pinpointed nutrient substitution and heavy metals content of algae biofertilizer as critical areas for further research if the performance of nutrient recovery systems such as TRENS is to be better characterized. Our study provides valuable feedback to the TRENS developers and identifies the importance of system expansion to include impacts outside the immediate nutrient recovery system itself. The study also show for the first time the successful evaluation of urban-to-agricultural water systems in EASETECH.

Improved Productivity of Neutral Lipids in Chlorella sp. A2 by Minimal Nitrogen Supply.

Nitrogen starvation is an efficient environmental pressure for increasing lipid accumulation in microalgae, but it could also significantly lower the biomass productivity, resulting in lower lipid productivity. In this study, green alga Chlorella sp. A2 was cultivated by using a minimal nitrogen supply strategy under both laboratory and outdoor cultivation conditions to evaluate biomass accumulation and lipid production. Results showed that minimal nitrogen supply could promote neutral lipid accumulation of Chlorella sp. A2 without a significant negative effect on cell growth. In laboratory cultivation mode, alga cells cultured with 18 mg L(-1) d(-1) urea addition could generate 74 and 416% (w/w) more neutral lipid productivity than cells cultured with regular BG11 and nitrogen starvation media, respectively. In outdoor cultivation mode, lipid productivity of cells cultured with 18 mg L(-1) d(-1) urea addition is approximately 10 and 88% higher than the one with regular BG11 and nitrogen starvation media, respectively. Notably, the results of photosynthetic analysis clarified that minimal nitrogen supply reduced the loss of photosynthetic capacity to keep CO2 fixation during photosynthesis for biomass production. The minimal nitrogen supply strategy for microalgae cultivation could promote neutral lipid accumulation without a significant negative effect on cell growth, resulting in a significant improvement in the lipid productivity.

Enhanced lipid accumulation of green microalga Scenedesmus sp. by metal ions and EDTA addition.

Effects of Fe(3+) (0-0.12 g/L), Mg(2+) (0-0.73 g/L) and Ca(2+) (0-0.98 g/L) on the biomass and lipid accumulation of heterotrophic microalgae were investigated in dark environment. The biomass and lipid production exhibited an increasing trend with increasing the concentrations of metal ions. In cultures with 1.2 × 10(-3) g/L Fe(3+), 7.3 × 10(-3) g/L Mg(2+) and 9.8 × 10(-4) g/L Ca(2+), the maximum biomass, total lipid content and lipid productivity reached 3.49 g/L, 47.4% and 275.7 mg/L/d, respectively. More importantly, EDTA addition (1.0 × 10(-3) g/L) could enhance the solubility of metal ions (iron and calcium) and increase their availability by microalgae, which evidently promote the lipid accumulation. Compared with the control, the total lipid content and lipid productivity increased 28.2% and 29.7%, respectively. These show that appropriate concentrations of metal ions and EDTA in the culture medium were beneficial to lipid accumulation of heterotrophic Scenedesmus sp. cells.

A novel flat plate algal bioreactor with horizontal baffles: structural optimization and cultivation performance.

A novel flat plate photobioreactor with horizontal baffles was developed. The effects of aeration intensity and aeration site were investigated using computational fluid dynamics. The effects of baffle structural parameters, including the ratio of clearance between the baffle and the bioreactor wall (d) to bioreactor width (D) and the ratio of distance between two adjacent baffles (h) to D on the flow and mixing performance were assessed. A good light/dark cycle and strong, uniform mixing performance were created at d/D=0.5 and h/D=1 when the aeration site was located at the bottom center. The cultivation performance was assessed by culturing Chlorella vulgaris 31. The maximum biomass productivity in the optimized bioreactor was 1.88 times than that of a traditional bioreactor without baffles. For a light path length of 80 mm, the optimized baffles offer a large economical advantage in improving algal productivity and reduce growth condition difference.

Enhanced energy conversion efficiency from high strength synthetic organic wastewater by sequential dark fermentative hydrogen production and algal lipid accumulation.

A two-stage process of sequential dark fermentative hydrogen production and microalgal cultivation was applied to enhance the energy conversion efficiency from high strength synthetic organic wastewater. Ethanol fermentation bacterium Ethanoligenens harbinense B49 was used as hydrogen producer, and the energy conversion efficiency and chemical oxygen demand (COD) removal efficiency reached 18.6% and 28.3% in dark fermentation. Acetate was the main soluble product in dark fermentative effluent, which was further utilized by microalga Scenedesmus sp. R-16. The final algal biomass concentration reached 1.98gL(-1), and the algal biomass was rich in lipid (40.9%) and low in protein (23.3%) and carbohydrate (11.9%). Compared with single dark fermentation stage, the energy conversion efficiency and COD removal efficiency of two-stage system remarkably increased 101% and 131%, respectively. This research provides a new approach for efficient energy production and wastewater treatment using a two-stage process combining dark fermentation and algal cultivation.