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Additional file1: Figure S1 DGGE fingerprints of bacterial 16S rRNA gene fragments from total community DNA. Figure S2 Fungal DGGE fingerprints of ITS fragments.

Additional file 1: Table S1. Molecules identified in chromatograms of ORG 1-3 fractions at different Retention Times. Figure S1. Chromatograms of organo-soluble fractions (ORG 1-3) solubilized during the Humeomic fractionation.

The present study, was aimed to evaluate the use of new sustainable and integrate agricultural practices, alternatively to the conventional and unsustainable use of mineral fertilizers and pesticides, developing new technologies able to understand how these interact with plants metabolism. Specifically, were tested bio-fertilizers based on the use of recycling products like composts, alone or in combination with plant bio-stimulants like microorganisms and humic substances. Such bio-effectros, frequently used in the organic farming, not only ensures food safety and increases soil biodiversity but are considered totally eco-sustainable. The comprehension of rhizospheric micro-bio-humeome interactions and the effects on plants metabolome was studied by Metabolomics through the use of advanced analytical techniques, like gas-chromatography coupled with mass spectrometry (GC-MS) and Nuclear Magnetic Resonance spectroscopy (NMR). The results underline a positive effects on plants, exerted by the combined use of bio-fertilizers and bio-stimulants, suggesting therefore, their use as valuable alternative to the conventional agricultural practices able to promote the environmental sustainability and production of healthy foods maintaining crop productivity. Additionally, in order to better understand witch molecular class of compound play a specific role in the micro-bio-humeome interaction, the Humeomics approach applied directly on materials enrich of HS, was firstly evaluated, representing a preliminary experiment with the future aim to apply this technique also on materials containing humo-like substances such as compost from different sources. The employment of analytical approach such as Metabolomics and Humeomics for understand the rhizospheric micro-bio-humeome interaction, appear necessary in order to formulate in the future, tailored prepared with specific bio-stimulats actions as a function of different crops and soils characteristics.

The molecular interaction of cocoa polyphenols with milk proteins were investigated in vitro by combined proteomic and biochemical strategies. Mass spectrometry and antioxidant activity assays allowed monitoring the binding of casein and whey protein fractions to cocoa polyphenols. In particular, the nature of interaction of beta-lactoglobulin (beta-Lg) with catechin and epicatechin was characterized and the amino acid residue at the binding site was identified. On the other side, antioxidant activity assays also showed a significant effect of the various milk protein fractions in decreasing the in vitro antioxidant activity of polyphenols, suggesting the existence of other types of protein-polyphenol interactions, probably weaker non-covalent bonds. From a nutritional point of view, these data indicate that the beta-Lg covalent modification by polyphenol alone do not support the hypothesis of a decrease in the bioavailability of polyphenols themselves (Scalbert & Williamson, 2000). This might also explain the maintenance of the antioxidant properties of cocoa polyphenols in cocoa-based beverages. These results suggest the perspective use of the model system developed to study other complex food matrices.