Characterization of a microalga Chlorella sp. well adapted to highly concentrated municipal wastewater for nutrient removal and biodiesel production

The feasibility of growing Chlorella sp. in the centrate, a highly concentrated municipal wastewater stream generated from activated sludge thickening process, for simultaneous wastewater treatment and energy production was tested. The characteristics of algal growth, biodiesel production, wastewater nutrient removal and the viability of scale-up and the stability of continuous operation were examined. Two culture media, namely autoclaved centrate (AC) and raw centrate (RC) were used for comparison. The results showed that by the end of a 14-day batch culture, algae could remove ammonia, total nitrogen, total phosphorus, and chemical oxygen demand (COD) by 93.9%, 89.1%, 80.9%, and 90.8%, respectively from raw centrate, and the fatty acid methyl ester (FAME) content was 11.04% of dry biomass providing a biodiesel yield of 0.12 g-biodiesel/L-algae culture solution. The system could be successfully scaled up, and continuously operated at 50% daily harvesting rate, providing a net biomass productivity of 0.92 g-algae/(L day).     

Culture of Scenedesmus sp. LX1 in the modified effluent of a wastewater treatment plant of an electric factory by photo-membrane bioreactor

To investigate the coupled technology for advanced wastewater treatment and microalgal biomass production, a photo-membrane bioreactor was constructed. The microalga Scenedesmus sp. LX1 was cultured in the bioreactor using liquor prepared from the effluent of an electronic device factory. The algal cell growth, nitrate nitrogen removal, orthophosphate phosphorus removal were investigated. When cultured with batch operation, the average specific growth rate was about 0.09 d⁻¹, and low nitrogen (N), phosphorus (P) concentrations in the liquor were achieved. However, under continuous operation with an inflow of 60 L h⁻¹, the average specific growth rate was only 0.02 d⁻¹, and removal rates of 100% for orthophosphate P and 46% for nitrate N were achieved. With the inflow of 120 L h⁻¹, the accumulated metal ions in the bioreactor adversely affected the algal cells. The algal cells were much easier to settle, and the removal efficiency for N and P decreased.     

Isolation and characterization of a Scenedesmus acutus strain to be used for bioremediation of urban wastewater

Green microalgae, due to their short growth cycle and to their ability to photosynthetically fix carbon dioxide producing an oil-rich biomass, have been proposed as an attractive alternative feedstock for the production of “second generation” biofuels. However, it has been anticipated that owing to their ability to colonize very different environments characterized by high levels of nitrogen, they can also be good candidates for bioremediation, thus integrating environmental protection with sustainable biomass production. We have isolated a strain belonging to Scenedesmus genus from urban wastewater. This isolate, Scenedesmus acutus PVUW12, was tested for its ability to grow and actively deplete eutrophicating inorganic molecules present in wastewater. In order to test its biomass productivity, the PVUW12 strain was grown in a vertical-column photobioreactor using standard growth medium obtaining a maximal productivity of 0.3?g dry weight L^?1?d. When the same strain was grown in the photobioreactor filled with wastewater collected from the final step of the local urban purifier plant containing 18.8?mg?L^?1 nitrate, we observed complete nitrogen removal coupled with a biomass production of about 0.74?g dry weight L^?1 within 3 days. After 10?days, the recovered biomass was analyzed for triglyceride content which was found to be 9.3% of the dry biomass. However, when algal cells were left for additional 10?days in static conditions the triglyceride content increased to 28.8%. These data show that this Scenedesmus strain can be used for wastewater bioremediation producing a biomass suitable for energy production.     

Growth characteristics of Botryococcus braunii 765 under high CO2 concentration in photobioreactor

To understand the potential of cultivating Botryococcus braunii with flue gas (normally containing high CO₂) for biofuel production, growth characteristics of B. braunii 765 with 2-20% CO₂ aeration were investigated. The results showed that the strain could grow well without any obvious inhibition under all tested CO₂ concentrations with an aeration rate of 0.2 vvm, even without any culture pH adjustment (ranged from 6.0 to 8.0). The maximum biomass among all conditions was 2.31 g L⁻¹ on 25th day at 20% CO₂. Hydrocarbon content and algal colony size increased with the increase of CO₂ concentration. A negative correlation between algal biomass and culture total phosphorus was observed (from −0.828 to −0.911, P < 0.01). Additionally, 2% sodium hypochlorite solution was used for photobioreactor sterilization to cultivate B. braunii.     

城市生活废水用于产油微藻培养

将废水与产油微藻培养结合起来,可以实现废水的无害化处理,还可为微藻的培养提供营养组分和大量水源。利用高产油栅藻,以城市生活废水为水源,在气泡柱式光反应器中,考察了添加不同营养组分对栅藻细胞的生长、生物质产量、总脂含量以及氮磷的去除情况的影响。结果表明：生活废水非常适合于产油微藻的培养,利用生活废水进行微藻培养中,仅需补充添加无机氮、无机磷、柠檬酸铁铵以及微量元素。但这些营养组分的加入量对藻细胞的生长、生物量和油脂积累有重要影响。在优化的废水培养基中微藻细胞浓度可达8.0 g/L左右,远高于标准BG11培养基5.0 g/L的水平。微藻细胞对于无机氮与磷有着高的吸收能力,在废水中加入185.25 mg/L以下无机氮,16.1 mg/L以下无机磷的条件下培养3~4 d后,培养液水体中未检测到有氮磷残留。由此表明利用城市生活废水培养含油微藻可以在获得微藻油脂产品的同时实现水体的无害化处理。     

Seasonal productivity of a periphytic algal community for biofuel feedstock generation and nutrient treatment

Algal biomass is a promising feedstock for biofuel production. With a high lipid content and high rate of production, algae can produce more oil on less land than traditional bioenergy crops. Algal communities can also be used to remove nutrients from impacted waters. The purpose of this study was to demonstrate the ability of an algal turf scrubber (ATS)™ to facilitate the growth of periphytic algal communities for the production of biomass feedstock and the removal of nutrients from a local stream. A pilot-scale ATS was implemented in Springdale, AR, and operated over the course of a nine-month sampling period. System productivity over the nine-month operating time averaged 26 g m⁻² d⁻¹. Total phosphorus and total nitrogen removal averaged 48% and 13%, respectively. The system showed potential for biomass generation and nutrient removal across three seasons.     

Competition between Salvinia minima and Spirodela polyrhiza mediated by nutrient levels and herbivory

We investigated the effects of initial biomass, nutrients, herbivory, and competition with Spirodela polyrhiza (L.) Schleid on Salvinia minima Baker biomass and density. S. minima populations were subjected to two levels of herbivory (control vs. two adults per plant) from the weevil Cyrtobagous salviniae Calder and Sands and eight levels of competition from S. polyrhiza, while growing in high (5 mg N l⁻¹) or low (0.5 mg N l⁻¹) nutrient conditions. Herbivory was the most important factor in S. minima biomass production while competition or fertility had no measurable impact. In contrast, S. polyrhiza biomass was mediated primarily by nutrients, not competition. There was no herbivory treatment for this plant. S. polyrhiza was superior to S. minima at converting nutrients to biomass but this did not give it a competitive advantage since S. minima biomass was unchanged when herbivory was absent. S. minima can generally overtop S. polyrhiza which, in turn, can form multiple layers within its mat. These characteristics may act to lessen competition between these species, thereby permitting their habitat sharing.     

Chitin: a potential new alternative nitrogen source for the tertiary,algal-based treatment of pulp and paper mill wastewater

Every day, pulp and paper mills in the USA discharge millions of liters of wastewater. Primary and secondary treatment of this wastewater often enriches it with phosphorus, resulting in uncontrolled eutrophication of receiving water bodies. A new method of tertiary wastewater treatment uses controlled growth of algae in a photobioreactor to sequester phosphorus into algal biomass, which is then harvested. This typically requires addition of a nitrogen fertilizer (nitrate, ammonium, or urea) to the water. We show on the laboratory scale that chitin can be used as an alternative source of nitrogen for the tertiary treatment of pulp mill wastewater using algae. We demonstrate that phosphorus can be efficiently removed from pulp wastewater using algae and chitin. Furthermore, phosphorus removal with chitin did not result in an increase in dissolved nitrogen in the wastewater because it is insoluble, unlike conventional nitrogen fertilizers. Despite its insolubility, it has recently been found that many diverse algae and cyanobacteria can use it as a source of nitrogen. Chitin has many advantages over conventional nitrogen fertilizers for use in wastewater treatment technologies. It is the second-most abundant natural polymer and is a waste product of the shellfish industry. Chitin is sustainable, inexpensive, and carbon neutral. Thus, chitin improves the sustainability and carbon footprints associated with water treatment, while the production of commercially attractive algal biomass helps to offset costs associated with the water treatment system itself.     

Nitrogen removal from wastewater with a low COD/N ratio at a low oxygen concentration

The goal of the study was to determine the effectiveness of nitrification and denitrification and the kinetics of ammonia removal from a mixture of wastewater and anaerobic sludge digester supernatant in an SBR at limited oxygen concentration. In addition, the COD removal efficiency and sludge production were assessed.In the SBR cycle alternating aerobic and anaerobic phases occurred; in the aeration phase the dissolved oxygen (DO) concentration was below 0.7 mg O₂/L. The low DO concentration did not inhibit ammonia oxidation-nitrification and the efficiency was ca. 96-98%. However, a relatively high COD concentration in the effluent was detected. The values of K_m and V_max, calculated from the Michaelis-Menten equation, were 43 mg N-NH₄/L and 15.64 mg N-NH₄/L h, respectively. Activated sludge production was almost stable (0.62-0.66 g MLVSS/g COD). A high net biomass production resulted from a low specific biomass decay rate of 0.0015 d⁻¹.     

Effects of light intensity and magnesium supplementation in pretreatment cycle on ammonium removal from wastewater of photobioreactor using a biofilter composed of the aerial microalga Trentepohlia aurea

Application of a laboratory-scale photobioreactor containing a biofilter composed of the aerial microalga Trentepohlia aurea to the removal of ammonium from synthetic wastewater was assessed to determine whether the system could be applied to water purification and the treatment of eutrophic water. The removal efficiency of the photobioreactor was tested after ten biofilter sheets (total dry weight cells: 50 mg) were cycled in nitrogen-free Bold’s basal (BB) medium for 72 h (pretreatment cycle). The ammonium removal ability was significantly enhanced when the photobioreactor was operated after performing the pretreatment cycle using nitrogen-free BB medium supplemented with magnesium. Moreover, the illumination conditions during the treatment were shown to affect the nitrogen removal ability, and this ability was strongly dependent on the concentrations of organic compounds (e.g., α-ketoglutarate and pyruvate) for assimilating the nitrogen source in the T. aurea biofilter.     

Nutrient removal of effluent from quail farm through cultivation of Wolffia arrhiza

The objective of this work was to study the nutrient removal using the Wolffiaarrhiza during the treatment of laying quails farm effluent. The relationship between W. arrhiza biomass and treatment time, the change in water qualities, and nitrogen-balance (N-balance) were evaluated. The results showed that a biomass of 12.0 g of W. arrhiza per liter of effluent and a treatment period of 30 days were found to provide the best conditions for W. arrhiza’s growth and the quality of the treated effluent in terms of biological oxygen demand, suspended solids, total phosphorus, nitrate, total ammonia nitrogen and total Kjeldahl nitrogen. The pH and salinity were similar for each level of biomass. The W. arrhiza biomasses of 4.00-12.0 g/l of effluent were suitable for W. arrhiza survival over time. Since W. arrhiza can fix N in the atmosphere, it can grow very well in effluent containing a low level of N.     

Assessment of carotenoid production by Dunaliella salina in different culture systems and operation regimes

The effect of operation regime and culture system on carotenoid productivity by the halotolerant alga Dunaliella salina has been analyzed. Operation strategies tested included batch and semi continuous regime, as well as a two-stage approach run simultaneously in both, open tanks and closed reactor. The best results were obtained with the closed tubular photobioreactor. The highest carotenoid production (328.8 mg carotenoid l⁻¹ culture per month) was achieved with this culture system operated following the two-stage strategy. Also, closed tubular photobioreactor provided the highest carotenoid contents (10% of dry weight) in Dunaliella biomass and β-carotene abundance (90% of total carotenoids) as well as the highest 9-cis to all-trans β-carotene isomer ratio (1.5 at sunrise).     

Hydrogen production by Rhodopseudomonas palustris WP 3-5 in a serial photobioreactor fed with hydrogen fermentation effluent

In this study, a lab-scale serial photobioreactor composed of three column reactors was constructed and continuously operated to investigate several parameters influencing photohydrogen production when using the synthetic wastewater and the anaerobic hydrogen fermentation effluents as the influents. The results indicated that better hydrogen production rate was obtained when the serial photobioreactor was operated under cellular recycling at a short HRT of 8 h. The serial photobioreactor maintained high hydrogen content ca. 80% in the produced gas and 0.4× dilution ratio was the suitable ratio for hydrogen production. When the photobioreactor fed with the real wastewater (Effluent 1) containing 100 mg/L NH₄Cl, Column 1 reactor successfully reduced ammonia concentration to about 60 mg/L for cell synthesis, resulting in a steady hydrogen production in the following two column reactors. The average hydrogen production rate was 205 mL-H₂/L/d.     

Biomass production and nitrogen and phosphorus removal by the green alga Neochloris oleoabundans in simulated wastewater and secondary municipal wastewater effluent

Biomass productivity of 350 mg DCW L⁻¹ day⁻¹ with a final biomass concentration of 3.15 g DCW L⁻¹ was obtained with Neochloris oleoabundans grown in artificial wastewater at sodium nitrate and phosphate concentrations of 140 and 47 mg L⁻¹, respectively, with undetectable levels of residual N and P in effluents. In secondary municipal wastewater effluents enriched with 70 mg N L⁻¹, the alga achieved a final biomass concentration of 2.1 g DCW L⁻¹ and a biomass productivity of 233.3 mg DCW L⁻¹ day⁻¹. While N removal was very sensitive to N:P ratio, P removal was independent of N:P ratio in the tested range. These results indicate that N. oleoabundans could potentially be employed for combined biofuel production and wastewater treatment.     

Nitrogen removal efficiencies in a free water surface constructed wetland in relation to plant coverage

Vegetation coverage is considered to be a key factor controlling nitrogen removal in wetlands. We describe the use of newly designed stainless steel incubation chambers to detect shifts in the in situ nitrate reduction activities associated to areas covered with common reed (Phragmites australis) and cattail (Typha latifolia) in the sediment of a free water surface constructed wetland (FWS-CW). Activities were measured at six different positions and times of the year and were related to physicochemical and hydraulic variables. Mean nitrate + nitrite reduction activities varied from 11.1 to 69.4 mg N/m²/h and showed a high variability within sediment types. Ammonification rates accounted for roughly 10% of the total nitrate reduction and were especially relevant in vegetated areas. Measured activities were highly above total nitrogen removal efficiencies estimated in the three parallel treatment cells of the Empuriabrava FWS-CW, indicating the potentiality of the system. In situ nitrate reduction activities correlated well with physichochemical characteristics such as pH and temperature. Additionally, differences in the total nitrogen removal efficiencies were detected between the three treatment cells and were related to changes in the water retention time. The plant species effect was detected in treatment cells of comparable hydraulic loads in which vegetation belts dominated by Typha latifolia were shown to have greater nitrogen removal efficiencies.     

Aerobic granulation with low strength wastewater at low aeration rate in A/O/A SBR reactor

The formation of aerobic granules with low organic loading synthetic wastewater (150-200 mg L⁻¹ of influent COD, acetate/propionate = 1/3) at low aeration rate (0.6 cm s⁻¹ of superficial gas velocity) had been investigated in the anaerobic/oxic/anoxic SBR. Aerobic granules with smooth surface and compact structure were successfully obtained after 50 days. However, these aerobic granules were unstable when the d(0.9) of granules increased to more than 1 mm. The results suggested that the aerobic granules with small diameter (smaller than 1000 μm) were more favorable for treating the low substrate loading wastewater at the low aeration rate. The cycle test revealed that most of the influent COD was removed at the anaerobic stage. The effluent concentrations of N-NH₄⁺ and P-PO₄³⁻ were lower than 1 mg L⁻¹, and the effluent concentration of nitrate gradually decreased with the granulation. Phosphate accumulating organisms were found to utilize O₂ or NO_x⁻ as electron acceptor for phosphorus removal in the study. Simultaneous nitrogen and phosphorus removal occurred inside the granules.     

Kinetic exploration of nitrate-accumulating microalgae for nutrient recovery

Within sustainable resource management, the recovery of nitrogen and phosphorus nutrients from waste streams is becoming increasingly important. Although the use of microalgae has been described extensively in environmental biotechnology, the potential of nitrate-accumulating microalgae for nutrient recovery has not been investigated yet. The ability of these marine microorganisms to concentrate environmental nitrate within their biomass is remarkable. The aim of this study was to investigate the application potential of nitrate-accumulating diatoms for nutrient recovery from marine wastewaters. The intracellular nitrate storage capacity was quantified for six marine diatom strains in synthetic wastewater. Amphora coffeaeformis and Phaeodactylum tricornutum stored the highest amount of nitrate with respectively 3.15 and 2.10 g N L^?1 of cell volume, which accounted for 17.3 and 4.6 %, respectively, of the total nitrogen content. The growth and nitrate and phosphate uptake of both diatoms were further analyzed and based on these features P. tricornutum showed the highest potential for nutrient recovery. A mathematical model was developed which included intracellular nitrate storage and the kinetic parameters were derived for P. tricornutum. Furthermore, a simulation study was performed to compare the performance of a proposed microalgal nutrient recovery unit with a conventional denitrification system for marine wastewater treatment. Overall, this study demonstrates the potential application of P. tricornutum for saline wastewater treatment with concurrent nitrogen and phosphorus recycling.     

Heterologous expression of Anabaena PCC 7120 all3940 (a Dps family gene) protects Escherichia coli from nutrient limitation and abiotic stresses

This study presents first hand data on the cloning and heterologous expression of Anabaena PCC 7120 all3940 (a dps family gene) in combating nutrients limitation and multiple abiotic stresses. The Escherichia coli transformed with pGEX-5X-2-all3940 construct when subjected to iron, carbon, nitrogen, phosphorus limitation and carbofuron, copper, UV-B, heat, salt and cadmium stress registered significant increase in growth over the cells transformed with empty vector under iron (0%), carbon (0.05%), nitrogen (3.7 mM) and phosphorus (2 mM) limitation and carbofuron (0.025 mg ml⁻¹), CuCl₂ (1 mM), UV-B (10 min), heat (47 °C), NaCl (6% w/v) and CdCl₂ (4 mM) stress. Enhanced expression of all3940 gene measured by semi-quantitative RT-PCR at different time points under above mentioned treatments clearly demonstrates its role in tolerance against aforesaid abiotic stresses. This study opens the gate for developing transgenic cyanobacteria capable of growing successfully under above mentioned stresses.