Enhanced washing of polycyclic aromatic hydrocarbons from contaminated soils by the empowered surfactant properties of de novo O-alkylated humic matter

Aqueous solutions of humic acid (HA) derivatized by a catalyzed O-alkylation reaction with methyl, pentyl, and benzyl groups at 40, 60, and 80% of total HA acidity were used to wash off polycyclic aromatic hydrocarbons (PAHs) from two contaminated soils. The enhanced surfactant properties enabled the alkylated HA to remove phenanthrene, anthracene, fluoranthene, and pyrene from both soils more extensively than the original unmodified HA, the 60% benzylation generally showing the greatest soil washing efficiency. For both soils, all alkylated HA revealed greater PAH removals than Triton X-100 nonionic surfactant, while the benzylated and methylated HA nearly and fully matched pollutants release by the anionic SDS in the coarse- and fine-textured soils, respectively. A consecutive second washing with 60% benzylated HA removed additional PAHs, in respect to the first washing, from the coarser-textured soil, except for fluoranthene, while removal from the finer-textured soil incremented even more for all PAHs. These findings indicate that the enhanced hydrophobicity obtained by a simple and unexpensive chemical derivatization of a natural humic surfactant can be usefully exploited in the washing of polluted soils, without being toxic to the soil biota and by potentially promoting the subsequent bio-attenuation of organic pollutants

Optimized procedure for the determination of P species in soil by liquid-state 31 P-NMR spectroscopy

Liquid-state 31P-NMR spectroscopy becomes progressively an important role for studying phosphorus (P) dynamics in soil. Soils of different origin and organic matter content were used to optimize sample preparation and re-dissolution procedures to improve characterization of P species in soil by 31P-NMR spectroscopy. The efficiency of P extraction from an untreated fresh soil was compared to that from freeze-dried and air-dried soil samples. A freeze-drying pretreatment not only provided the greatest extraction yields of total and organic P from both farmland and forest soils but also enhanced the intensity of signals for inorganic and organic P species in 31P-NMR spectra, except for polyphosphates. Re-dissolution of freeze-dried soil extracts in relatively dilute alkaline solution and addition of a small aliquot of concentrated HCl to the NMR tube prior to analysis improved the quality of NMR spectra. Finally, the visibility of relatively weak P signals, such as for phosphorus diesters, phosphonates, polyphosphate, phospholipids, and DNA were reproducibly enhanced when 31P-NMR spectra were generated after at least 15 h of acquisition time. The optimized procedure presented here ensured the greatest detectability of inorganic and organic P species by liquid-state P-NMR spectroscopy in soil extracts

Combination of humic biostimulants with a microbial inoculum improves lettuce productivity, nutrient uptake, and primary and secondary metabolism

Biostimulants of natural origin represent a growing ecological strategy to increase crops productivity, especially when applied in combination with microbial bioeffectors. We studied the effect of biostimulants such as Potassium Humates (KH) from Leonardite and Compost Tea (CT) from green compost on both productivity and nutritional status of lettuce plants, as well as on the primary and secondary metabolism of treated plants, when amended either alone or in combination with a commercial microbial inoculum (M+), mainly based on arbuscular mycorrhizal fungi (Micosat TabPlus)

Antibacterial and antioxidant properties of humic substances from composted agricultural biomasses

Background: Bioactive components isolated from composted agricultural biomasses have been receiving progressive attention, because they may improve the antibiotic susceptibility of drug resistant bacterial strains. Here, three different humic substances (HS) were isolated from composted artichoke (HS-CYN) and pepper (HS-PEP) wastes, and from coffee grounds (HS-COF), and characterized by infrared spectrometry, NMR spectroscopy, thermochemolysis–GC/MS, and high-performance size-exclusion chromatography. The antibacterial activity of HS was evaluated against some pathogenic bacterial strains, while their bioactivity was determined by a germination assay on basil (Red–Violet variety) seeds. Results: HS-CYN and HS-PEP exhibited the largest antioxidant activity and most significant antimicrobial capacity against some gram-positive bacterial strains, such as Staphylococcus aureus and Enterococcus faecalis. The same HS determined a significant increase of both root and epicotyls in seed germination experiments. The bioactivity of HS was related not only to their specific molecular composition but also to the conformational stability of their suprastructures. Specifically, the greatest bioactive and antimicrobial properties were related to the largest abundance of hydrophobic aromatic and phenolic components and to a more rigid conformational arrangement, that, in turn, appeared to be related to a small fragmentation degree of lignin structures. Conclusions: Our results showed that extraction of bioactive HS from green composts may be a sustainable and eco-compatible way to valorise agricultural byproducts. HS may be indeed exploited as substrates to produce novel materials not only to improve plant productivity but also for medical applications. Graphical Abstract: [Figure not available: see fulltext.

Fertilisation with compost mitigates salt stress in tomato by affecting plant metabolomics and nutritional profiles

Abstract Background Salinity is one of the major threats for crop growth and yield and its rate of expansion is expected to increase. We conducted a pot experiment to evaluate and compare the effect of a green compost addition and mineral fertilisation, on growth, nutrition and metabolites of tomato plants, exposed to increasing doses of NaCl. Results Although the development of stressed plants was lower than the corresponding controls, compost-treated plants performed better than mineral-amended plants watered with the same amount of salt. The different plant growth was related to an increased nutritional status. Namely, compost-treated plants showed a larger content of macro- and micronutrients, and a greater accumulation of osmoprotectants, such as soluble sugars and amino acids. Moreover, compost-treated plants showed a larger content of metabolites involved in modulating the response to salt stress, such as molecules related to energy transfer in plants and precursors of Reactive Oxygen Species scavenging compounds. Overall, the better performance of compost-added plants may be attributed to a greater availability of the organic forms of nutrients and to the positive bioactivity of compost-derived humic substances. Conclusions Compost application efficiently mitigated salt stress in tomato plants in respect to mineral fertilisation. This alleviating role was associated to the induction of a more efficient metabolic response that increased accumulation of metabolites involved in modulating the salinity stress. Therefore, fertilising with composted agricultural residue may represent a convenient alternative to mineral fertilisers to grow tomato plants in the presence of salt stress. Graphical Abstrac

Potential of three microbial bio-effectors to promote maize growth and nutrient acquisition from alternative phosphorous fertilizers in contrasting soils

Background Agricultural production is challenged by the limitation of non-renewable resources. Alternative fertilizers are promoted but they often have a lower availability of key macronutrients, especially phosphorus (P). Biological inoculants, the so-called bio-effectors (BEs), may be combined with these fertilizers to improve the nutrient use efficiency. Methods The goal of this study was to assess the potential of three BEs in combination with alternative fertilizers (e.g., composted manure, biogas digestate, green compost) to promote plant growth and nutrient uptake in soils typical for various European regions. Pot experiments were conducted in Czech Republic, Denmark, Germany, Italy, and Switzerland where the same variety of maize was grown in local soils deficient in P in combination with alternative fertilizers and the same set of BEs (Trichoderma, Pseudomonas, and Bacillus strains). Common guidelines for pot experiment implementation and performance were developed to allow data comparison, and soils were analyzed by the same laboratory. Results Efficiency of BEs to improve maize growth and nutrient uptake differed strongly according to soil properties and fertilizer combined. Promising results were mostly obtained with BEs in combination with organic fertilizers such as composted animal manures, fresh digestate of organic wastes, and sewage sludge. In only one experiment, the nutrient use efficiency of mineral recycling fertilizers was improved by BE inoculation. Conclusions These BE effects are to a large extent due to improved root growth and P mobilization via accelerated mineralization

Triple Blockade of Ido-1, PD-L1 and MEK as a Potential Therapeutic Strategy in NSCLC

BACKGROUND: Despite the recent progress in the treatment and outcome of Non Small Cell Lung Cancer (NSCLC), immunotherapy has still significant limitations reporting a significant proportion of patients not benefiting from therapy, even in patients with high PD-L1 expression. We have previously demonstrated that the combined inhibition of MEK and PD-L1 in NSCLC patients derived three dimensional cultures exerted significant synergistic effect in terms of immune-dependent cancer cell death. However, subsequent experiments analyzing the expression of Indoleamine 2,3-dioxygenase-1 (Ido-1) gene expression demonstrated that Ido-1 resulted unaffected by the MEK inhibition and even increased after the combined inhibition of MEK and PD-L1 thus representing a potential escape mechanism to this combination. METHODS: We analyzed transcriptomic profile of NSCLC lung adenocarcinoma cohort of TCGA (The Cancer Genome Atlas), stratifying tumors based on EMT (Epithelial mesenchymal Transition) score; in parallel, we investigated the activation of Ido-1 pathway and modulation of immune cytokines productions both in NSCLC cells lines, in peripheral blood mononuclear cells (PBMCs) and in ex-vivo NSCLC spheroids induced by triple inhibition with an anti-PD-L1 monoclonal antibody, the MEK inhibitor and the Ido-1 inhibitor. RESULTS: In NSCLC lung adenocarcinoma patient cohort (from TCGA) Ido-1 gene expression was significantly higher in samples classified as mesenchymal according EMT score. Similarly, on a selected panel of NSCLC cell lines higher expression of MEK and Ido-1 related genes was detected in cells with mesenchymal phenotype according EMT score, thus suggesting a potential correlation of co-activation of these two pathways in the context of EMT, with cancer cells sustaining an immune-suppressive microenvironment. While exerting an antitumor activity, the dual blockade of MEK and PD-L1 enhances the secretion of pro-inflammatory cytokines (IFNγ, TNFα, IL-12 and IL-6) and, consequently, the expression of new immune checkpoints such as Ido-1. The triple inhibition with an anti-PD-L1 monoclonal antibody, the MEK inhibitor and the Ido-1 inhibitor demonstrated significant antiproliferative and proapoptotic activity on ex-vivo NSCLC samples; at the same time the triple combination kept increased the levels of pro-inflammatory cytokines produced by both PBMCs and tumor spheroids in order to sustain the immune response and simultaneously decreased the expression of other checkpoint (such as CTLA-4, Ido-1 and TIM-3) thus promoting an immune-reactive and inflamed micro-environment. CONCLUSIONS: We show that Ido-1 activation is a possible escape mechanism to immune-mediated cell death induced by combination of PD-L1 and MEK inhibitors: also, we show that triple combination of anti-PD-L1, anti-MEK and anti-Ido-1 drugs may overcome this negative feedback and restore anti-tumor immune response in NSCLC patients\u27 derived three dimensional cultures

Effectiveness of bio-effectors on maize, wheat and tomato performance and phosphorus acquisition from greenhouse to field scales in Europe and Israel: a meta-analysis

Biostimulants (Bio-effectors, BEs) comprise plant growth-promoting microorganisms and active natural substances that promote plant nutrient-acquisition, stress resilience, growth, crop quality and yield. Unfortunately, the effectiveness of BEs, particularly under field conditions, appears highly variable and poorly quantified. Using random model meta-analyses tools, we summarize the effects of 107 BE treatments on the performance of major crops, mainly conducted within the EU-funded project BIOFECTOR with a focus on phosphorus (P) nutrition, over five years. Our analyses comprised 94 controlled pot and 47 field experiments under different geoclimatic conditions, with variable stress levels across European countries and Israel. The results show an average growth/yield increase by 9.3% (n=945), with substantial differences between crops (tomato > maize > wheat) and growth conditions (controlled nursery + field (Seed germination and nursery under controlled conditions and young plants transplanted to the field) > controlled > field). Average crop growth responses were independent of BE type, P fertilizer type, soil pH and plant-available soil P (water-P, Olsen-P or Calcium acetate lactate-P). BE effectiveness profited from manure and other organic fertilizers, increasing soil pH and presence of abiotic stresses (cold, drought/heat or salinity). Systematic meta-studies based on published literature commonly face the inherent problem of publication bias where the most suspected form is the selective publication of statistically significant results. In this meta-analysis, however, the results obtained from all experiments within the project are included. Therefore, it is free of publication bias. In contrast to reviews of published literature, our unique study design is based on a common standardized protocol which applies to all experiments conducted within the project to reduce sources of variability. Based on data of crop growth, yield and P acquisition, we conclude that application of BEs can save fertilizer resources in the future, but the efficiency of BE application depends on cropping systems and environments

Novel fertilising products from lignin and its derivatives to enhance plant development and increase the sustainability of crop production

Lignin is a by-product of biorefineries and paper mills and is usually discarded or burnt. However, it may represent a source of novel fertilising products, able to support the sustainable intensification of agricultural productions. The aim of this review is to explore the literature regarding the effect of lignin application towards plant growth and nutrient use efficiency. First, we reviewed the biostimulant role of lignin, which was reported to positively perturbing plant hormonal balances or improving the efficiency of photosynthesis and respiration. Also, when added to soils, lignin was shown to enhance nitrogen uptake, as well as the development of beneficial soil microorganisms. Then, we summarised the research related to the chemical modifications of lignin structure devised to boost its bioactivity, an approach opening possibilities to tailor the effects of lignin addition on plant development. We further examined the literature about the use of lignin and its derivatives as starting substrates to produce sustainable materials (chelates, coatings, micro- and nano-materials) for the slow release of plant nutrients. Encouraging results emerged from the summarised articles, suggesting that lignin may replace the currently used synthetic polymers exploited to chelate or entrapping nutrients. We additionally hereby highlighted the role of lignin chemical nature in affecting its biological and release properties, hence pointing out the relevance of thoroughly studying its structure at a molecular level by the most advanced analytical tools. Finally, we suggested the need for researchers to combine their skills and expertise, in order to develop more efficient lignin-inspired fertilisers