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  • Bioprocess to produce biostimulants/biofertilizers based on microalgae grown using piggery wastewater as nutrient source
    Publication . Ferreira, Alice; Corrêa, D. O.; Ribeiro, Belina; Silva, Teresa Lopes da; Santos, Cláudia Marques dos; Acién, F. Gabriel; Gouveia, Luisa
    ABSTRACT: 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.
  • Innovative approach in sustainable agriculture: Harnessing microalgae potential via subcritical water extraction
    Publication . Ferreira, Alice; Vladic, Jelena; Corrêa, D. O.; Butzke, Valéria Louzada Leal; Martins, Pedro L.; Ribeiro, Belina; Santos, Cláudia Marques dos; Acién, F. Gabriel; Gouveia, Luisa
    ABSTRACT: 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.
  • Unlocking the potential of Euglena gracilis cultivated in piggery wastewater: biomass production, nutrient removal, and biostimulant potential in lettuce and tomato plants
    Publication . Butzke, Valéria Louzada Leal; Ferreira, Alice; Corrêa, D. O.; Furlan, Júnior Mendes; Gouveia, Luisa; Schneider, Rosana C. S.; Corbellini, Valeriano Antonio
    ABSTRACT: Microalgae are increasingly recognized as a valuable resource for bolstering sustainability in agriculture. However, current research and patents primarily focus on Chlorella spp., Scenedesmus spp., and Spirulina spp., thus leaving the vast diversity of microalgae relatively unexplored for agricultural applications. Euglena gracilis (Euglenophyta) is a microalga renowned for its resilience to diverse environmental stressors and capability to produce a variety of bioactive metabolites. This study investigated the potential of cultivating E. gracilis in piggery wastewater for nutrient recycling and as a source of beneficial biomolecules, particularly for biostimulant use. Utilizing raw wastewater diluted to 25% (P25) and pre-treated wastewater with photo-Fenton (PF), the research found that E. gracilis exhibited elevated cell density, biomass concentration, and overall cell health in both wastewaters compared to a synthetic medium (BG11-NPK). This was due to its efficient removal of nutrients, especially ammoniacal-nitrogen and phosphate, resulting in a biomass rich in polyunsaturated fatty acids, amino acids, and paramylon content. The whole-cell biomass significantly enhanced the germination index of lettuce and tomato seeds compared to the water control. Additionally, it promoted cell expansion and root formation in cucumber cotyledons, exhibiting similarities to phytohormones such as gibberellin, cytokinin, and auxin. Furthermore, it is suggested that E. gracilis biomass contains molecules related to resistance to environmental stresses, particularly in tomatoes, given the enhancement in the seedling vigor index. E. gracilis exhibited remarkable adaptability to piggery wastewater, recycling nutrients and yielding biomass rich in bioactive molecules with potential as plant biostimulants. These findings significantly contribute to understanding E. gracilis's potential applications in agriculture and developing a circular bioeconomy.
  • Design optimisation of five pilot-scale two-stage vertical flow-constructed wetlands for piggery wastewater treatment
    Publication . Karan, N.; Gogoi, Jayanta; Ganguly, Anasuya; Brito, António; Marques dos Santos, C.; de Oliveira Corrêa, Diego; Gouveia, Luisa; Mutnuri, Srikanth
    ABSTRACT: With growing pig farming, sustainable piggery wastewater treatment methods are essential for environmental protection. This study evaluated five pilot-scale two-stage vertical flow-constructed wetlands (VFCWs) with varying configurations of aeration, plantation, and saturation zones. Three VFCW configurations (1VFCW, 2VFCW, and 3VFCW) were unsaturated, while 4VFCW and 5VFCW were saturated in the second stage (up to 60 and 90 cm, respectively). The 5VFCW featured a stacked configuration with no space between its two stages. Passive aeration was selectively applied in 2VFCW, 3VFCW, 4VFCW, and 5VFCW, while plants were present in most configurations except the control. Saturated 4VFCW achieved the highest removal efficiency for TN (77.03 ± 16.24%) and NO3− (46.06 ± 45.96%), while the stacked 5VFCW showed the highest removal for chemical oxygen demand (COD) (94.17 ± 4.85%) and Total ammoniacal nitrogen (TOC) (86.35 ± 6.78%). Unsaturated 1VFCW excelled in TAN removal (98.89 ± 0.33%), and the control system (C) showed the highest removal efficiency for PO43− (90.38 ± 6.52%) and TOC (87.52 ± 9.83%). Overall, 4VFCW emerged as the most balanced and effective system, supported by an optimal combination of aerobic and anaerobic conditions that facilitated sequential nitrification and denitrification, along with an extended hydraulic retention time due to saturation.