Integrating microalgae-based wastewater treatment, biostimulant production, and hydroponic cultivation: a sustainable approach to water management and crop production.

A natural appearing microalgae-bacteria consortium was used to process urban wastewater. The process was done in an 80 m2 raceway reactor and the results were compared to an identical reactor operated using freshwater supplemented with commercial fertilisers. The biomass harvesting was done using commercial ultrafiltration membranes to reduce the volume of culture centrifuged. The membrane allowed achieving a biomass concentration of ∼9-10 g L-1. The process proposed avoids the use of centrifuges and the drying of the biomass, two of the most energy consuming steps of conventional processes. The specific growth rate in freshwater and the wastewater-based media was estimated as 0.30 ± 0.05 and 0.24 ± 0.02 days-1, respectively (p < 0.05). The maximum concentration reached at the end of the batch phase was 0.96 ± 0.03 and 0.83 ± 0.07 g L-1 when the biomass was produced using freshwater and wastewater, respectively (p < 0.05). The total nitrogen removal capacity of the system was on average 1.35 g m-2·day-1; nitrogen assimilation into biomass represented 60%-95% of this value. Furthermore, the P-PO4 3- removal capacity of the system varied from 0.15 to 0.68 g m-2·day-1. The outlet effluent of the reactor was used as a nutrient source in the hydroponic production of zucchini seedlings, leading to an increase in the root dry weight and the stem diameter compared to the water alone. The produced biomass showed potential for use as feedstock to produce plant biostimulants with positive effects on root development and chlorophyll retention.

Effect of seawater on the biomass composition of Spirulina produced at a pilot-scale.

The microalga Arthrospira platensis BEA 005B was produced in 11.4 m3 raceway photobioreactors and a culture medium based on commercial fertilisers and either freshwater or seawater. The biomass productivity of the reactors operated at a fixed dilution rate of 0.3 day-1 decreased from 22.9 g·m-2·day-1 when operated using freshwater to 16.3 g·m-2·day-1 when the biomass was produced using seawater. The protein content of the biomass produced in seawater was lower; however, the content of essential amino acids including valine, leucine and isoleucine was higher. Seawater also triggered the production of carotenoids and altered the synthesis and accumulation of fatty acids. For example, the biomass produced using seawater showed a 319% and 210% higher content of oleic and eicosenoic acid, respectively. The results demonstrate that it is possible to produce the selected microalga using seawater after an adaptation period and that the composition of the produced biomass is suitable for food applications.

Agricultural products from algal biomass grown in piggery wastewater: A techno-economic analysis.

The intensification of livestock activities lead to an increase in waste generation with high content of nutrients, as is the case of piggery wastewater. However, this type of residue can be used as culture media for algae cultivation in thin-layer cascade photobioreactors to reduce its environment impact and produce a valorizable algal biomass. Biostimulants were produced by enzymatic hydrolysis and ultrasonication of microalgal biomass, using membranes (Scenario 1) or centrifugation (Scenario 2) as harvesting methods. The co-production of biopesticides by solvent extraction was also evaluated using membranes (Scenario 3) or centrifugation (Scenario 4). The four scenarios were analyzed by a technoeconomic assessment estimating the total annualized equivalent cost and the production cost, i.e., the minimum selling price. Centrifugation provided biostimulants approximately 4 times more concentrated than membranes, but with higher expense due to the cost of the centrifuge (contribution of 62.2 % in scenario 2) and the electricity requirements. The biopesticide production resulted the highest contribution to investment cost in scenarios 3 and 4 (34 % and 43 % respectively). The use of membranes was also more advantageous to produce biopesticides, although it was 5 times more diluted than using centrifuge. The biostimulant production cost was 65.5 €/m3 with membranes and 342.6 €/m3 by centrifugation and the biopesticide production cost was 353.7 €/m3 in scenario 3 and 2,122.1 €/m3 in scenario 4. Comparing the treatment of 1 ha of land, the cost of the biostimulant produced in the four scenarios was lower than the commercial one (48.1 %, 22.1 %, 45.1 % and 24.2 % respectively). Finally, using membranes for biomass harvesting allowed economically viable plants with lower capacity and longer distance for biostimulant distribution (up to 300 km) than centrifuge (188 km). The algal biomass valorization for agricultural products production is an environmentally and economically feasible process with the adequate capacity of the plant and distribution distance.

Production of enzymatic hydrolysates with in vitro antioxidant, antihypertensive, and antidiabetic properties from proteins derived from Arthrospira platensis.

The microalga Arthrospira platensis BEA 005B was produced using 80 m2 (9 m3) raceway photobioreactors achieving a biomass productivity of 28.2 g·m-2·day-1 when operating the reactors in semi-continuous mode (0.33 day-1). The produced biomass was rich in proteins (58.1 g·100 g-1) and carbohydrates (25.6 g·100 g-1); the content of phycocyanins and allophycocyanins was 115.4 and 36.9 mg·g-1, respectively. Ultrasounds and high-pressure homogenisation allowed recovering approximately 90% of the initial protein content of the biomass; however, the energetic requirements of the former (∼100 kJ·kg-1) were significantly lower than those of high-pressure homogenisation (∼200 kJ·kg-1). An in silico analysis revealed that papain and ficin would allow releasing a large number of bioactive peptides with antioxidant, antihypertensive (ACE-I and renin), and antidiabetic (DPP-IV, α-amylase, and α-glucosidase) properties. Both were assessed in vitro together with Alcalase and pepsin leading to the generation of enzymatic hydrolysates with in vitro bioactivity.

Long-term assessment of the nutrient recovery capacity and biomass productivity of Scenedesmus almeriensis in raceway reactors using unprocessed urban wastewater.

The present work aims to assess the treatment of unprocessed urban wastewater using the microalga Scenedesmus almeriensis. Two 12 m3 raceway reactors, one supplemented by wastewater and the second by chemical fertilizer, operating outdoors in a semi-continuous mode, were used for eight months. Results suggested that S. almeriensis can be produced in wastewater without affecting the photosynthetic apparatus reaching a productivity of 13 g·m-2·day-1 on average in both the systems. Furthermore, the nutrient content in terms of nitrogen, phosphorous and chemical oxygen demand of the wastewater was reduced under the European limitations during most of the period, with an average removal rate of 2.2, 0.2 and 3.0 g·m-2·day-1 respectively. Therefore, raceways demonstrated a high potential for microalgal production and successful biotreatment, proving robust and reliable. Finally, the effect of environmental conditions on biomass productivity of the clean system was evaluated in a model with high accuracy (R2 = 0.9, p = 0.0002).

Biostimulant Capacity of Chlorella and Chlamydopodium Species Produced Using Wastewater and Centrate.

The aim of the present study was to assess the potential of producing four microalgal strains using secondary-treated urban wastewater supplemented with centrate, and to evaluate the biostimulant effects of several microalgal extracts obtained using water and sonication. Four strains were studied: Chlorella vulgaris UAL-1, Chlorella sp. UAL-2, Chlorella vulgaris UAL-3, and Chlamydopodium fusiforme UAL-4. The highest biomass productivity was found for C. fusiforme, with a value of 0.38 ± 0.01 g·L-1·day-1. C. vulgaris UAL-1 achieved a biomass productivity of 0.31 ± 0.03 g·L-1·day-1 (the highest for the Chlorella genus), while the N-NH4+, N-NO3-, and P-PO43- removal capacities of this strain were 51.9 ± 2.4, 0.8 ± 0.1, and 5.7 ± 0.3 mg·L-1·day-1, respectively. C. vulgaris UAL-1 showed the greatest potential for use as a biostimulant-when used at a concentration of 0.1 g·L-1, it increased the germination index of watercress seeds by 3.5%. At concentrations of 0.5 and 2.0 g·L-1, the biomass from this microalga promoted adventitious root formation in soybean seeds by 220% and 493%, respectively. The cucumber expansion test suggested a cytokinin-like effect from C. vulgaris UAL-1; it was also the only strain that promoted the formation of chlorophylls in wheat leaves. Overall, the results of the present study suggest the potential of producing C. vulgaris UAL-1 using centrate and wastewater as well as the potential utilisation of its biomass to develop high-value biostimulants.

Biorefinery Approach Applied to the Production of Food Colourants and Biostimulants from Oscillatoria sp.

In this study, a biorefinery based on Oscillatoria sp. is developed to produce high-value compounds such as C-phycocyanin, used in food colourant applications, and biostimulants, used in agriculture-related applications. First, the Oscillatoria biomass production was optimized at a pilot scale in an open raceway reactor, with biomass productivities equivalent to 52 t/ha·year being achieved using regular fertilizers as the nutrient source. The biomass produced contained 0.5% C-phycocyanins, 95% of which were obtained after freeze-thawing and extraction at pH 6.5 and ionic strength (FI) 100 mM, with a purity ratio of 0.71 achieved in the final extract. This purity ratio allows for use of the extract directly as a food colourant. Then, the extract's colourant capacity on different beverages was evaluated. The results confirm that C-phycocyanin concentrations ranging from 22 to 106 mg/L produce colours similar to commercial products, thus avoiding the need for synthetic colourants. The colour remained stable for up to 12 days. Moreover, the safety of the extracted C-phycocyanin was confirmed through toxicity tests. The waste biomass was evaluated for use as a biostimulant, with the results confirming a relevant auxin-like positive effect. Finally, an economic analysis was conducted to evaluate different scenarios. The results confirm that the production of both C-phycocyanin and biostimulants is the best scenario from an economic standpoint. Therefore, the developed biomass processing scheme provides an opportunity to expand the range of commercial applications for microalgae-related processes.

Optimisation of Operational Conditions during the Production of Arthrospira platensis Using Pilot-Scale Raceway Reactors, Protein Extraction, and Assessment of their Techno-Functional Properties.

The aim of the present study was to identify the optimum combination of dilution rate and depth of the culture to maximise the Arthrospira platensis BEA005B (Spirulina) productivity using 80 m2 raceway reactors. By varying these two main operational conditions, the areal biomass productivity of the reactors varied by over 55%. The optimum combination, optimised using a surface response methodology, was a depth of 0.10 m and a dilution rate of 0.33 day-1, which led to a biomass productivity of 30.2 g·m-2·day-1 on a dry weight basis when operating the reactors in semi-continuous mode. The composition of the produced biomass was 62.2% proteins, 42.5% carbohydrates, 11.6% ashes, and 8.1% lipids. The isolated proteins contained all the essential amino acids (except for tryptophan, which was not determined); highlighting the content of valine (6.8%), histidine (8.3%), and lysine (7.5%). The functional properties of the proteins were also assessed, demonstrating huge potential for their use in the development of innovative and sustainable foods.

Microalgae for the Food Industry: From Biomass Production to the Development of Functional Foods.

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Characterisation of the volatile profile of microalgae and cyanobacteria using solid-phase microextraction followed by gas chromatography coupled to mass spectrometry.

Microalgae and microalgae-derived ingredients are one of the top trends in the food industry. However, consumers' acceptance and purchase intention of a product will be largely affected by odour and flavour. Surprisingly, the scientific literature present a very limited number of studies on the volatile composition of microalgae and cyanobacteria. In order to fill the gap, the main objective of the present study was to elucidate the volatile composition of seven microalgal and cyanobacterial strains from marine and freshwaters, with interest for the food industry while establishing its potential impact in odour. Among the seven selected strains, Arthrospira platensis showed the highest abundance and chemical diversity of volatile organic compounds (VOCs). Aldehydes, ketones, and alcohols were the families with the highest diversity of individual compounds, except in Arthrospira platensis and Scenedesmus almeriensis that showed a profile dominated by branched hydrocarbons. Marine strains presented a higher abundance of sulfur compounds than freshwater strains, while the ketones individual profile seemed to be more related to the taxonomical domain. The results of this study indicate that the VOCs composition is mainly driven by the individual strain although some volatile profile characteristics could be influenced by both environmental and taxonomical factors.