Dynamic performance of combined biochar from co-pyrolysis of pig manure with invasive weed: Effect of natural aging on Pb and As mobilization in polluted mining soil.

Due to the natural biochar aging, the improvement of soil quality and immobilization of soil pollutants achieved by biochar may change; understanding the dynamic evolution of the in situ performance of biochar in these roles is essential to discuss the long-term sustainability of biochar remediation. Therefore, in this study, combined biochar from co-pyrolysis of pig manure and invasive Japanese knotweed - P1J1, as well as pure pig manure - PM - and pure Japanese knotweed - JK - derived biochar were applied to investigate their remediation performance in a high As- and Pb-polluted soil with prolonged incubation periods (up to 360 days). Biochar application, especially P1J1 and PM, initially promoted soil pH, dissolved organic carbon, and EC, but the improvements were not constant through time. The JK-treated soil exhibited the highest increase of soil organic matter (OM), followed by P1J1 and then PM, and OM did not change with aging. Biochar, especially P1J1, was a comprehensive nutrient source of Ca, K, Mg, and P to improve soil fertility. However, while soluble cationic Ca, K, and Mg increased with time, anionic P decreased over time, indicating that continuous P availability might not be guaranteed with the aging process. The total microorganism content declined with time; adding biochars slowed down this tendency, which was more remarkable at the later incubation stage. Biochar significantly impeded soil Pb mobility but mobilized soil As, especially in PM- and P1J1-treated soils. However, mobilized As gradually re-fixed in the long run; meanwhile, the excellent Pb immobilization achieved by biochars was slightly reduced with time. The findings of this study offer fresh insights into the alterations in metal(loid)s mobility over an extended duration, suggesting that the potential mobilization risk of As is reduced while Pb mobility slightly increases over time.

Impact of soil treatment with Nitrilo Triacetic Acid (NTA) on Cd fractionation and microbial biomass in cultivated and uncultivated calcareous soil.

Purpose: The aim of this study was to evaluate the effectiveness of nitrilotriacetic acid (NTA) on cadmium (Cd) fractions and microbial biomass in a calcareous soil spiked with Cd under cultivated (Zea mays L.) and uncultivated regime subject to soil leaching condition. Expanding investigations related to soil-plant interactions on metal-contaminated soils with insights on microbial activity and associated soil toxicity perspective provides novel perspectives on using metal-chelating agents for soil remediation. Methods: The experimental factors were three levels of Cd contamination (0, 25, and 50 mg kg-1 soil) and three levels of NTA (0, 15, and 30 mmol L-1) in loamy soil under maize-cultured and non-cultured conditions. During the experiment, the adding NTA and leaching processes were performed three times. Results: The results showed that the amount of leached Cd decreased in cultivated soil compared to uncultivated soil due to partial uptake of soluble Cd by plant roots and changes in Cd fractions in soil, so that Cd leached in Cd50NTA30 was 9.2 and 6.1 mg L-1, respectively, in uncultivated and cultivated soils. Also, Cd leached in Cd25NTA30 was 5.7 and 3.1 mg L-1 respectively, in uncultivated and cultivated soils. The best treatment in terms of chemical and microbial characteristics of the soil with the high percentage of Cd removed from the soil was Cd25NTA30 in cultivated soil. In Cd25NTA30 compared to Cd25NTA0 in cultivated soil, pH (0.25 unit), microbial biomass carbon (MBC, 65.0 mg kg-1), and soil respiration (27.5 mg C-CO2 kg-1 24 h-1) decreased, while metabolic quotient (qCO2, 0.05) and dissolved organic carbon (DOC, 20.0 mg L-1) increased. Moreover, the changes of Cd fractions in Cd25NTA30 in cultivated soil compared to uncultivated soil were as follows; the exchangeable Cd (F1, 0.27 mg kg-1) and Fe/Mn-oxide-bounded Cd (F4, 0.15 mg kg-1) fractions increased, in contrast, carbonate-Cd (F2, 2.67 mg kg-1) and, organically bounded Cd (F3, 0.06 mg kg-1) fractions decreased. NTA had no significant effect on the residual fraction (F5). Conclusion: The use of NTA, especially in calcareous soils, where most of the Cd is bound to calcium carbonate, was able to successfully convert insoluble fractions of Cd into soluble forms and increase the removal efficiency of Cd in the phytoremediation method. NTA is a non-toxic chelating agent to improve the accumulation of Cd in maize.

Metal behavior and soil quality changes induced by the application of tailor-made combined biochar: An investigation at pore water scale.

The remediation performance of biochar varies based on the biomass used for its production. Further innovation involves developing tailor-made biochar by combining different raw materials to compensate for the limitations of pure biochar. Therefore, tailor-made combined biochar produced from the co-pyrolysis of pig manure and invasive Japanese knotweed (P1J1), as well as biochars produced from these feedstocks separately, i.e., pure pig manure (PM) and pure Japanese knotweed (JK), were applied to Pb and As contaminated soil to evaluate the biochar-induced changes on soil properties, microbial activity, DOM, and metal and metalloids solubility at the soil pore water scale. Biochar application reduced soluble Pb, whereas enhanced the As mobility; the increased soil pH after biochar addition played a fundamental role in reducing the Pb solubility, as revealed by their significant negative correlation (r = -0.990, p < 0.01). In contrast, the release of dissolved P strongly influenced As mobilization (r = 0.949, p < 0.01), especially in P-rich PM and P1J1 treatments, while JK showed a marginal effect in mobilizing As. Soils treated with PM, P1J1, and JK mainly increased Gram-negative bacteria by 56 %, 52 %, and 50 %, respectively, compared to the control. Fluorescence excitation-emission matrix spectroscopy combined with parallel factor analysis identified three components in pore water DOM, C1 (long wavelength humic-like), C2 (short wavelength humic-like), and C3 (protein-like), which were dominant respectively in the P1J1, JK, and PM-added soil. A principal component analysis (PCA) confirmed that the PM and P1J1 had similar performance and were more associated with releasing P and Mg and specific DOM components (C1 and C3). Meanwhile, P1J1 supplemented soil OM/OC and K, similar to JK. The results of this study suggest that combined biochar P1J1 can comprehensively enhance soil quality, embodying the advantages of pure PM and JK biochar while overcoming their shortcomings.

Improving biochar properties by co-pyrolysis of pig manure with bio-invasive weed for use as the soil amendment.

Over recent years, pyrolysis has grown into a mature technology with added value for producing soil improvers. Further innovations of this technology lie in developing tailor-made products from specific feedstocks (or mixtures thereof) in combination with adjusted mixing ratio-temperature regimes. In this context, co-pyrolysis of pig manure (PM) and the invasive plant Japanese knotweed (JK) at different mixture ratios (w/w) of 3:1 (P3J1), 1:1 (P1J1), and 1:3 (P1J3) and varying temperatures (400-700 °C) was studied to address the low carbon properties and heavy metals (HMs) risks of manure-derive biochars and beneficially ameliorate the bio-invasion situation by creating value from the plant biomass. Co-pyrolysis of PM with JK increased by nearly 1.5 folds the fixed carbon contents in the combined feedstock biochars obtained at 600 °C compared with PM-derived biochar alone, and all combined feedstock biochars met the requirements for soil improvement and carbon sequestration. The total HMs in PM biochars were significantly reduced by adding JK. The combined feedstock biochar P1J1 generated at 600 °C was the most effective in transforming Cu and Zn into more stable forms, accordingly reducing the associated environmental risk of heavy metal leaching from the biochar. In addition, the accumulation of macronutrients can be an added benefit of the co-pyrolysis process, and P1J1-600 was also the biochar that retained the most nutrients (P, Ca, Mg, and K).

The Impact of Salt Accumulation on the Growth of Duckweed in a Continuous System for Pig Manure Treatment.

Duckweed (Lemna) is a possible solution for the treatment of aqueous waste streams and the simultaneous provision of protein-rich biomass. Nitrification-Denitrification effluent (NDNE) from pig manure treatment has been previously used as a growing medium for duckweed. This study investigated the use of a continuous duckweed cultivation system to treat NDNE as a stand-alone technology. For this purpose, a system with a continuous supply of waste streams from the pig manure treatment, continuous biomass production, and continuous discharge that meets the legal standards in Flanders (Belgium) was simulated for a 175-day growing season. In this simulation, salt accumulation was taken into account. To prevent accumulating salts from reaching a toxic concentration and consequently inhibiting growth, the cultivation system must be buffered, which can be achieved by altering the depth of the system. To determine the minimum depth of such a system, a tray experiment was set up. For that, salt accumulation data obtained from previous research were used for simulating systems with different pond depths. It was found that a depth of at least 1 m is needed to prevent a significant relative growth inhibition at the end of the growing season compared to the start. This implies a high water consumption (5-10 times more than maize). As a response, a second cultivation system was investigated for the use of more concentrated NDNE. For this purpose, salt tolerance experiments were conducted on synthetic and biological media. Surprisingly, it was observed that duckweed grows better on diluted NDNE (to 75% NDNE, or EC of 8 mS/cm) than on a synthetic medium (EC of 1.5 mS/cm), indicating the potential of such a system.

Biodegradation and effects of EDDS and NTA on Zn in soil solutions during phytoextraction by alfalfa in soils with three Zn levels.

In chelator-enhanced Zn phytoremediation studies, it is crucial to understand how the degradable chelators and the competition from other ions influence the concentration of Zn in soil solutions. This study investigated the biodegradability of two chelators (EDDS: Ethylenediamine-N,N'-disuccinic acid, and NTA: Nitrilotriacetic acid) and their effects on the Zn concentration in the soil solution during the growth of alfalfa (Medicago Sativa L.). The chelators were added at four doses (0, 0.5, 2 and 5 mmol kg-1) in soils with varying Zn levels (189, 265 and 1496 mg kg-1). The results showed that the lag phase before EDDS and NTA biodegradation varied from 0 to 7 days in the three soils. EDDS and NTA were completely decomposed within the assessed 57 days regardless of the applied dosage, with a half-life of 1.3-3.0 days in highly Zn-contaminated soil and 4.2-10.8 days in the two other soils. In soil solutions, the change in solubilized Zn was in line with EDDS and NTA degradation kinetics. Cu, Al, Fe and Mn were the main metal ions that competed against Zn for chelation. Besides, Ni competed with Zn in the whole process. Ca did not compete effectively in the three soils, while Mg was a competitor only at the initial stage. Our results show the importance of considering both the biodegradation rate and the competition between the target cation and other elements present in the soil when using chelators to enhance phytoremediation. A 30-day explorative incubation experiment is recommended to evaluate the appropriate application time of chelators and the target Zn exposure time for plants during phytoremediation.

Maximizing nutrient recycling from digestate for production of protein-rich microalgae for animal feed application.

The integration of phototrophic microalgal production and anaerobic digestion can recycle excess nutrients across European surplus hotspots to produce protein-rich biomass for nutritional applications. However, the challenging physico-chemical properties of raw digestate constrain microalgal growth and limit digestate valorization potential. This study focused on the pre-treatment of food waste-based digestate using paper-filtration to improve its properties for cultivating Desmodesmus sp. and Chlorella vulgaris. The microalgal growth performance in paper-filtered digestate (PFD, 10 µm-pore size) was then compared to growth in membrane-filtered digestate (MFD, 0.2 µm-pore size). A microplate-based screening coupled with Cytation device assessment of PFD and MFD samples after dilution and with/without phosphorus supplementation showed that PFD was the best substrate. Moreover, phosphorus supplementation resulted in improved growth at higher digestate concentrations (5-10% v/v PFD), indicating the importance of using a balanced growth medium to increase the volumetric usage of digestate. Results were validated in a 3-L bioreactor at 10% PFD with phosphorus supplementation, reaching a biomass concentration of 2.4 g L-1 with a protein and carbohydrate content of 67% and 13% w/w respectively. This trial indicates that paper-filtration is a promising pre-treatment technique to maximize digestate recycling and deliver a sustainable animal feed-grade protein alternative.

Cu phytoextraction and biomass utilization as essential trace element feed supplements for livestock.

Copper (Cu), as an essential element, is added to animal feed to stimulate growth and prevent disease. The forage crop alfalfa (Medicago sativa L.) produced during Cu phytoextraction may be considered a biofortified crop to substitute the Cu feed additives for livestock production, beneficially alleviating Cu contamination in soils and reducing its input into agriculture systems. To assess this, alfalfa was grown in three similar soils with different Cu levels, i.e., 11, 439 and 779 mg kg-1 for uncontaminated soil (A), moderately Cu-contaminated soil (B) and highly Cu-contaminated soil (C), respectively. EDDS (Ethylenediamine-N,N'-disuccinic acid) was applied to the soils seven days before the first cutting at four rates (0, 0.5, 2 and 5 mmol kg-1) to enhance bioavailable Cu uptake. Alfalfa grew well in soils A and B but not in the highly Cu-contaminated soil. After applying EDDS, a significant biomass reduction of the first cutting shoot was only observed with 5 mmol kg-1 EDDS in the highly Cu-contaminated soil, with a 45% (P < 0.05) decrease when compared to the control. Alfalfa grown in the three soils gradually wilted after the first cutting with 5 mmol kg-1 EDDS, and Cu concentrations in the first cutting shoot were augmented strongly, by 250% (P < 0.05), 3500% (P < 0.05) and 6700% (P < 0.05) compared to the controls, respectively. Cu concentrations in alfalfa shoots were found to be higher in this study than in some fodder plants and further augmented in soils with higher Cu levels and with EDDS application. These findings suggest that alfalfa grown on clean soils or soils with up to 450 mg Cu kg-1 (with appropriate EDDS dosages) has the potential to be considered as a partial Cu supplementation for livestock. This research laid the foundation for the integration between Cu-phytoextraction and Cu-biofortification for livestock.

Zn phytoextraction and recycling of alfalfa biomass as potential Zn-biofortified feed crop.

Zn is an essential micronutrient for living organisms and, in that capacity, it is added to animal feed in intensive livestock production to promote growth and eliminate diseases. Alfalfa (Medicago sativa L.) may have the potential to compensate and substitute the need for chemical Zn additives in feeds as a Zn-biofortified feed crop when grown on Zn-enriched soils. Thus, this possibility was investigated with a greenhouse experiment using three soils with Zn concentrations (mg kg-1) of 189 (soil A), 265 (soil B) and 1496 (soil C). Ethylenediamine-N,N'-disuccinate acid (EDDS) and Nitrilotriacetic acid (NTA) at different rates (0 as control, 0.5, 2 and 5 mmol kg-1) were applied as soil additives to enhance the phytoextraction efficiency of alfalfa. The results showed that Zn was highly transferable in alfalfa tissues in the three soils even without additives. EDDS was more effective than NTA in enhancing Zn phytoextraction by alfalfa. The maximum Zn accumulation in the third cutting shoots was obtained with the EDDS concentration of 5 mmol kg-1 in soil A and of 2 mmol kg-1 in soil B, with a 462% and 162% increase compared with controls, respectively. However, the higher EDDS concentration resulted in a significant reduction in biomass production. In soil C, all EDDS concentrations resulted in similar Zn accumulations in the third shoot. To improve the phytoextraction efficacy of Zn while minimizing its phytotoxicity on alfalfa, the rate of 2 mmol kg-1 EDDS proved to be optimal for soil B, and 0.5 mmol kg-1 EDDS for soils A and C. Findings suggest that phytoextraction of Zn-enriched soil can be combined with Zn biofortification, thus allowing to recycle Zn into biomass that can, to an extent, substitute Zn feed additives. This study provided a primary data set for the combination of Zn-biofortification and Zn-phytoextraction.

Effect of the growth medium composition on nitrate accumulation in the novel protein crop Lemna minor.

Duckweed is a potential alternative protein source for food and feed. However, little is known about the nitrate accumulation in this plant. A high nitrate level in vegetables can indirectly lead to an elevated intake of nitrites and N-nitroso compounds, increasing the risk of diseases for humans and animals. This research hypothesizes that the nitrate accumulation of Lemna minor differs between growing media. Additionally, it evaluates whether legal safety levels of nitrate for human and animal intake are exceeded. The duckweed was grown on (i) rainwater, and (ii) three synthetic media containing different nutrient levels. Furthermore, (iii) biological effluent of swine manure treatment and (iv) aquaculture effluent from pikeperch production were used, as these are potential media for closing nutrient loops in the agriculture sector. It was found that nitrate levels increased with the increasing availability of macronutrients in the water, and pH showed a particularly strong negative correlation with the nitrate levels in the plant. Nevertheless, nitrate content never exceeded 530 mg NO3 kg-1 fresh weight. To conclude, Lemna minor's nitrate content was below safety limits for human consumption in all tested growing media; however, a potential risk for ruminants was observed as these are more sensitive to nitrate conversions in their gastro-intestinal track.