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- Biostimulant and biopesticide potential of microalgae growing in piggery wastewaterPublication . Ferreira, Alice; Melkonyan, Lusine; Carapinha, Sofia; Ribeiro, Belina; Figueiredo, Daniel; Avetisova, Gayane; Gouveia, LuisaABSTRACT: Pig farming generates highly polluting wastewaters which entail serious environmental issues when not adequately managed. Microalgae systems can be promising for cost, energy and environment-efficient treatment of piggery wastewater (PWW). Aside from clean water, the produced biomass can be used as biostimulants and biopesticides contributing to a more sustainable agriculture. Three microalgae (Tetradesmus obliquus, Chlorella protothecoides, Chlorella vulgaris) and one cyanobacterium (Synechocystis sp.) were selected after a preliminary screening in diluted wastewater (1:20) to treat PWW. The nutrient removals were 62-79% for COD (chemical oxygen demand), 84-92% for TKN (total Kjeldahl nitrogen), 79-92% for NH4+ and over 96% for PO43−. T. obliquus and C. protothecoides were the most efficient ones. After treating PWW, the produced biomass, at 0.5 g L−1, was assessed as a biostimulant for seed germination, root/shoot growth, and pigment content for tomato, watercress, cucumber, soybean, wheat, and barley seeds. We observed an overall increase on germination index (GI) of microalgae-treated seeds, owing to the development of longer roots, especially in T. obliquus and C. vulgaris treatments. The microalgae treatments were especially effective in cucumber seeds (75-138% GI increase). The biopesticide activity against Fusarium oxysporum was also evaluated at 1, 2.5 and 5 g L−1 of microalgae culture. Except for Synechocystis sp., all the microalgae tested inhibited the fungus growth, with T. obliquus and C. vulgaris achieving inhibitions above 40% for all concentrations.
- Green approach for the valorization of microalgae Tetradesmus obliquusPublication . Gouveia, Luisa; Jazic, Jelena Molnar; Ferreira, Alice; Maletic, Snezana; Cvetkovic, Dragoljub; Vidovic, Senka; Vladic, JelenaABSTRACT: The main goal of this study was to develop an efficient, green approach for the valorization of Tetradesmus obliquus biomass, with zero waste. This microalga was selected because it is widespread, resistant, easy for cultivation, and fast-growing. In the first step, supercritical carbon dioxide (ScCO2) extraction followed by rapid gas decompression was used for the extraction of biomass. The following step was to apply ultrasound-assisted (UA), microwave-assisted (MA), and subcritical water (SW) extraction on the ScCO2-treated biomass to determine the most efficient processing technology. SW demonstrated to be a superior technique over MW and UA with regard to extraction yield and antioxidant content. Moreover, the chemical and microbiological profiles of SW extracts were determined to evaluate their potential and safety. In addition, to create a procedure with zero waste, the solid waste after SW extraction (residue) was analyzed. The organic profile of extracts and residues contained compounds that belong to groups of aliphatic saturated hydrocarbons, aliphatic unsaturated hydrocarbons, alkylated hydrocarbons, ketones, phenols, and esters. Furthermore, these compounds can be applied in different industries including the pharmaceutical and cosmetic industries. Additionally, the content of metals in residues indicated that this material can be used as animal feed and in agriculture. Finally, a complete reduction of microorganisms present in the initial biomass was obtained for the extracts and residues, indicating their safety.
- Bioprocess to produce biostimulants/biofertilizers based on microalgae grown using piggery wastewater as nutrient sourcePublication . Ferreira, Alice; Corrêa, D. O.; Ribeiro, Belina; Silva, Teresa Lopes da; Santos, Cláudia Marques dos; Acién, F. Gabriel; Gouveia, LuisaABSTRACT: In the present work, two downstream processes - high-pressure homogenization at 100 (HPH-100) and 1200 bar (HPH-1200), and enzymatic hydrolysis (EH) - were tested to produce biostimulant extracts from Tetradesmus obliquus grown in piggery wastewater at two concentrations (12.8 and 88.3 g/L). Extracts before and after centrifugation (C) were evaluated in four bioassays using garden cress (germination), mung bean (auxin-like activity), and cucumber (auxin- and cytokinin-like activity) relative to distilled water. The initial microalgal culture, without any treatment, had the best germination results (162 % at 0.2 g/L) and the only one that showed cytokinin-like activity (141 % at 0.5 g/L). In both auxin-like bioassays, the HPH-1200 + C and EH + C originated high values (186 and 155 % for cucumber, 290 and 285 % for mung bean, respectively). For mung bean, the HPH-1200 achieved the highest auxin-like effect (378 %). Finally, the extracted biomass contained essential nutrients for biofertilization, complementing the biostimulant extracts for sustainable agriculture application.
- Application of Green Technology to Extract Clean and Safe Bioactive Compounds from Tetradesmus obliquus Biomass Grown in Poultry WastewaterPublication . Vladic, Jelena; Jazic, Jelena Molnar; Ferreira, Alice; Maletic, Snezana; Cvetkovic, Dragoljub; Agbaba, Jasmina; Vidovic, Senka; Gouveia, LuisaABSTRACT: Microalgae are capable of assimilating nutrients from wastewater (WW), producing clean water and biomass rich in bioactive compounds that need to be recovered from inside the microalgal cell. This work investigated subcritical water (SW) extraction to collect high-value compounds from the microalga Tetradesmus obliquus after treating poultry WW. The treatment efficiency was evaluated in terms of total Kjeldahl nitrogen (TKN), phosphate, chemical oxygen demand (COD) and metals. T. obliquus was able to remove 77% TKN, 50% phosphate, 84% COD, and metals (48-89%) within legislation values. SW extraction was performed at 170 degrees C and 30 bar for 10 min. SW allowed the extraction of total phenols (1.073 mg GAE/mL extract) and total flavonoids (0.111 mg CAT/mL extract) with high antioxidant activity (IC50 value, 7.18 mu g/mL). The microalga was shown to be a source of organic compounds of commercial value (e.g., squalene). Finally, the SW conditions allowed the removal of pathogens and metals in the extracts and residues to values in accordance with legislation, assuring their safety for feed or agriculture applications.
- From piggery wastewater to wheat using microalgae towards zero wastePublication . Ferreira, Alice; Figueiredo, Daniel; Ferreira, Francisca; Marujo, Ana; Bastos, Carolina R. V.; Martin-Atanes, Guillermo; Ribeiro, Belina; Štěrbová, Karolína; Santos, Cláudia Marques dos; Acién, F. Gabriel; Gouveia, LuisaABSTRACT: Microalgae production is still expensive, driving the need to lower costs while strengthening the industry's environmental sustainability. Microalgae are recognized tools for efficient wastewater treatment, offering the recycling of nutrients and water for agriculture, and producing biomass rich in growth-promoting compounds to improve plant productivity and resistance to adverse conditions. The use of wastewater can reduce cultivation costs as it is a source of nutrients and water. Alternative low-cost methods can significantly decrease harvesting costs, which represents one of the most expensive steps of the whole process.The goal of this work was to evaluate the potential of wastewater-grown microalga biomass for agriculture purposes. To reduce production costs, the microalga Tetradesmus obliquus was produced in pre-treated photo-Fenton (PF) piggery wastewater in combination with the use of different harvesting techniques - electro-coagulation, flocculation, and centrifugation, and different combinations. From the wastewater treatment pro-cess, two fractions (biomass and supernatant) were evaluated for germination and growth of wheat (Triticum aestivum L.) plants and compared to non-harvested microalga culture (MC), distilled water, and Hoagland (synthetic) solution. The concentrated resulting from PF was also tested as a biofertilizer.The results confirm that both biomass and supernatants are useful for agricultural applications. The obtained biomass elicited a 20-105 % increase in germination index compared to the control, while supernatants were inhibiting. The opposite trend was observed at later stages of wheat growth, where the nutrient-enriched su-pernatants and the PF concentrate (PF-CC) increased the number of tillers (3-5) and leaves (30-42) after 83 days. Wheat plants treated with MC and PF-CC produced similar number of ears (3.4 & PLUSMN; 0.5 and 6.0 & PLUSMN; 4.1 ears per plant, respectively) than the synthetic control (5.7 & PLUSMN; 1.4) after 182 days. All fractions obtained from the process can be used in a zero-waste process.
- Nannochloropsis oceanica biomass enriched by electrocoagulation harvesting with promising agricultural applicationsPublication . Figueiredo, Daniel; Ferreira, Alice; Gama, Florinda; Gouveia, LuisaABSTRACT: Electrocoagulation is a promising technology to harvest and concentrate microalgae while saving costs on secondary dewatering steps. However, the sacrificial electrodes release salts that impact the media and the harvested biomass. This study evaluated the effects of Fe, Zn, and Mg electrodes on Nannochloropsis oceanica harvesting and elementary composition of biomass and supernatants. Moreover, plant bioavailability of electrocoagulation minerals attached to biomass was assessed in the tomato plant model Solanum lycopersicum (cv. ‘Cherry’). Fe electrodes had better performance at lower power consumption and operation costs, followed by Zn and Mg. Electrocoagulation changes biomass and supernatant nutrient composition. Electrodes precipitated Mg and Ca from the nutrient media, enriching N. oceanica biomass, but increased Pb 2–4 times and depleted P in supernatants. Finally, Fe and Mg electrode metals in the biomass were proven bioavailable to S. lycopersicum seedlings, making electrocoagulation harvested biomass a promising bioresource to agricultural applications.
- Combining biotechnology with circular bioeconomy: from poultry, swine, cattle, brewery, dairy and urban wastewaters to biohydrogenPublication . Ferreira, Alice; Marques, Paula; Ribeiro, Belina; Assemany, Paula; Vieira De Mendonça, Henrique; Barata, Ana; Oliveira, Ana Cristina; Reis, Alberto; Pinheiro, H. M.; Gouveia, LuisaABSTRACT: The ability of microalgae to grow in nutrient-rich environments and to accumulate nutrients from wastewaters (WW) makes them attractive for the sustainable and low-cost treatment of WW. The valuable biomass produced can be further used for the generation of bioenergy, animal feed, fertilizers, and biopolymers, among others. In this study, Scenedesmus obliquus was able to remove nutrients from different wastewaters (poultry, swine and cattle breeding, brewery and dairy industries, and urban) with removal ranges of 95-100% for nitrogen, 63-99% for phosphorus and 48-70% for chemical oxygen demand. The biomass productivity using wastewaters was higher (except for poultry) than in synthetic medium (Bristol), the highest value being obtained in brewery wastewater (1025 mg/(L.day) of freeze-dried biomass). The produced biomass contained 31-53% of proteins, 12-36% of sugars and 8-23% of lipids, regardless of the type of wastewater. The potential of the produced Scenedesmus obliquus biomass for the generation of BioH(2) through batch dark fermentation processes with Enterobacter aerogenes was evaluated. The obtained yields ranged, in mL H-2/g Volatile Solids (VS), from 50.1 for biomass from anaerobically digested cattle WW to 390 for swine WW, whereas the yield with biomass cultivated in Bristol medium was 57.6 mL H-2/gvs.
- Innovative approach in sustainable agriculture: Harnessing microalgae potential via subcritical water extractionPublication . Ferreira, Alice; Vladic, Jelena; Corrêa, D. O.; Butzke, Valéria Louzada Leal; Martins, Pedro L.; Ribeiro, Belina; Santos, Cláudia Marques dos; Acién, F. Gabriel; Gouveia, LuisaABSTRACT: Microalgae can contribute to sustainable agriculture and wastewater treatment. This study investigated Tetradesmus obliquus, grown in piggery wastewater (To-PWW), as a biostimulant/biofertilizer compared to biomass grown in synthetic medium (To-B). Subcritical water extraction was tested for disruption/hydrolysis of wet biomass, at three temperatures (120, 170, and 220 °C) and two biomass loads (1:10 and 1:80 (g dry biomass/mL water)). Extracts were evaluated for germination, and root formation/expansion. Residues were quantified for nutrient composition to assess their biofertilizer potential and tested for their affinity to oil compounds for bioremediation. The best germination was achieved by To-B extracts at 170 °C (1:10: 148 % at 0.2 g/L, 1:80: 145 % at 0.5 g/L). Only To-PWW extracts at 0.2 g/L had a significant germination effect (120 °C: 120–123 % for both loads; 170 °C: 115 % for 1:80). To-PWW extract at 120 °C and 1:10 significantly affected cucumber and mung bean root formation (224 and 268 %, respectively). Most extracts significantly enhanced root expansion, with all To-B extracts at 1:10 showing the best results (139–181 %). The residues contained essential nutrients (NPK), indicating their biofertilizer potential, helping decrease synthetic fertilizers demands. To-B residues had high affinity to toluene and diesel but lower to used cooking and car oils. To-PWW showed very low affinity to all oil compounds. Finally, all residues were only able to form stable emulsions with the used car oil. This study fully exploits the use of microalgal biomass in sustainable agriculture, producing biostimulant extracts, and residues for biofertilizer and bioremediation, from a low-cost wastewater source.
- Resources recovery from domestic wastewater by a combined process: anaerobic digestion and membrane photobioreactorPublication . Ferrera, Elvira; Ruigómez, Ignacio; Vela‑Basto, Carolina; Ferreira, Alice; Gouveia, Luisa; Vera, LuisaABSTRACT: Anaerobic and membrane technologies are a promising combination to decrease the energy consumption associated with wastewater treatment, allowing the recovery of resources: organic matter as biomethane, nutrient assimilation by microalgae and reclaimed water. In this study, domestic wastewater was treated using a combination of an upflow anaerobic sludge blanket sludge reactor (UASB) and a membrane photobioreactor (MPBR). The outdoor facilities were operated continuously for three months under unfavourable environmental conditions such as lack of temperature control, winter season with lower solar irradiation and lower daylight hours which was a challenge for the present work, not previously described. The energetic valorisation of the organic matter present in the wastewater by biomethane produced in the UASB would contribute to reducing overall facilities’ energy requirements. The ultrafiltration (UF) membrane facilitated the harvesting of biomass, operating at 10 L·h−1·m−2 during the experimental period. Although the main contribution to fouling was irreversible, chemical cleanings were not necessary due to effective fouling control, which prevented the final TMP from exceeding 25 kPa. In addition, microalgae-bacterial consortium developed without prior inoculation were harvested from the MPBR using membrane assistance. The obtained biomass was also successfully tested as a biostimulant for corn germination/growth, as well as a biopesticide against Rhizoctonia solani and Fusarium oxysporum.
- Biostimulant potential of Scenedesmus obliquus grown in brewery wastewaterPublication . Navarro-Lopez, Elvira; Ruiz-Nieto, Angela; Ferreira, Alice; Acien, Gabriel; Gouveia, LuisaABSTRACT: Microalgae are microorganisms with the capacity to contribute to the sustainable and healthy food production, in addition to wastewater treatment. The subject of this work was to determine the potential of Scenedesmus obliquus microalga grown in brewery wastewater to act as a plant biostimulant. The germination index of watercress seeds, as well as the auxin-like activity in mung bean and cucumber, and in the cytokinin-like activity in cucumber bioassays were used to evaluate the biostimulant potential. Several biomass processes were studied, such as centrifugation, ultrasonication and enzymatic hydrolysis, as well as the final concentration of microalgal extracts to determine their influence in the biostimulant activity of the Scenedesmus biomass. The results showed an increase of 40% on the germination index when using the biomass at 0.1 g/L, without any pre-treatment. For auxin-like activity, the best results (up to 60% with respect to control) were obtained at 0.5 g/L of biomass extract, after a combination of cell disruption, enzymatic hydrolysis and centrifugation. For cytokinin-like activity, the best results (up to 187.5% with respect to control) were achieved without cell disruption, after enzymatic hydrolysis and centrifugation at a biomass extract concentration of 2 g/L.
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