Literature search – Exploring in silico protein toxicity prediction methods to support the food and feed risk assessment

This report is the outcome of an EFSA procurement (NP/EFSA/GMO/2018/01) reviewing relevant scientific information on in silico prediction methods for protein toxicity, that could support the food and feed risk assessment. Several proteins are associated with adverse (toxic) effects in humans and animals, by a variety of mechanisms. These are produced by plants, animals and bacteria to prevail in hostile environments. In the present report, we present an integrated pipeline to perform a comprehensive literature and database search applied to proteins with toxic effects. \u201cToxin activity\u201d and \u201ctoxin-antitoxin system\u201d strings were used as inputs for this pipeline. UniProtKB was considered as the reference database, and only the UniProtKB curator-reviewed proteins were considered in the pipeline. Experimentally- determined structures and homology-based in silico 3D models were retrieved from protein structures repositories; family-, domain-, motif- and other molecular signature-related information was also obtained from specific databases which are part of the InterPro consortium. Protein aggregation associated with adverse effects was also investigated using different search strategies. This work can serve as the basis for further exploring novel risk assessment strategies for new proteins using in silico predictive methods

Effetti degli oli essenziali di agrumi applicati in post-emergenza sul controllo delle infestanti e sulla comunit\ue0 microbica del suolo

Sebbene gli erbicidi sintetici ad oggi siano il mezzo pi\uf9 utilizzato per il controllo delle erbe infestanti, \ue8 ampiamente dimostrato che essi hanno un impatto negativo sul suolo, sull'ambiente e, inoltre, iniziano a manifestarsi fenomeni di resistenza. Pertanto, \ue8 importante sviluppare mezzi alternativi che siano al contempo efficaci e rispettosi dell\u2019ambiente. Gli olii essenziali estratti dalle piante sono stati da sempre utilizzati per molteplici usi come agenti battericidi, virucidi, fungicidi, insetticidi, medicinali e cosmetici. Diversi studi hanno dimostrato tali attivit\ue0 degli oli essenziali estratti dagli agrumi, ma scarsi sono gli studi che hanno investigato il loro potenziale uso come erbicidi. Inoltre, anche se diversi studi hanno dimostrato un effetto antimicrobico, tali studi sono stati condotti prevalentemente in vitro e non in vivo. Obiettivo del presente lavoro \ue8 stato quello di valutare la potenzialit\ue0 degli oli essenziali estratti dalle bucce di limone e arancia i) per limitare lo sviluppo delle erbe infestanti e ii) il loro effetto sulla comunit\ue0 microbica del suolo. Gli oli essenziali testati sono stati estratti per idrodistillazione e con pressa a freddo: LI e AI, olio essenziale di limone e arancio, rispettivamente, estratti con pressa a freddo; LU e AU, olio essenziale di limone e arancio, rispettivamente, estratti tramite idrodistillazione. Ai fini del presente lavoro, il suolo utilizzato \ue8 stato prelevato presso un campo sperimentale dell'Universit\ue0 degli Studi di Palermo. Dopo il prelievo, il suolo \ue8 stato essiccato all'aria e setacciato con setaccio con fori del diametro di un centimetro. Successivamente, 500 grammi di suolo sono stati posti in 20 vaschette di alluminio (10 7 20 cm) ed incubati in condizioni naturali. Durante l\u2019incubazione il suolo \ue8 stato mantenuto al 50% della sua capacit\ue0 idrica di campo (WHC). Dopo l\u2019emergenza delle piantine, ovvero alla comparsa della terza foglia (ventesimo giorno dall\u2019inizio dell\u2019incubazione), ciascun olio essenziale \ue8 stato applicato nebulizzandolo sulle piantine, a giorni alterni per una settimana (tre applicazioni in totale). Per ogni vaso e per ogni trattamento, sono stati applicati 5 mL di una soluzione contenente 1 L di acqua, 10 mL di olio essenziale e 1 mL di emulsionante. Nel suolo di controllo la soluzione applicata alle piantine non conteneva olio essenziale. Dopo 15 giorni dall\u2019ultima applicazione dei trattamenti, i suoli sono stati campionati in modo totale, setacciati ed analizzati per la determinazione del carbonio della biomassa microbica (metodo fumigazione-estrazione), respirazione basale (emissione di CO2) e struttura della comunit\ue0 microbica (analisi degli acidi grassi dei fosfolipidi). Inoltre, per ogni trattamento sono stati determinati alcuni parametri delle piante come peso secco e fresco totale, numero di specie e numero di piante per specie. In questo lavoro vengono riportati e discussi i principali risultati

ATR-FTIR chemical screening for adulterants and sugar characterisation in honeys

The search for improvements in honey analysis is an important topic due to the relevance that adulterations and variations of sugar composition have in this food. Attenuated Total Reflectance Fourier Transform InfraRed spectroscopy (ATR-FTIR) is a reliable and fast analytical instrumentation for analysis of liquids, semi-solids and solids, which is currently used in food chemistry for qualitative and quantitative investigations. This work is aimed to build a fast analytical method, using ATR-FTIR and Partial Least Square (PLS) chemometric tool, to quantitative determination of glucose, fructose and sucrose present in honeys. We analyzed 25 honeys from Trentino and Sicily and a significant spectral range from the whole ATR-FTIR spectra were selected. Then a PLS multivariate calibration model was built by using mixtures of glucose, fructose and sucrose at various concentration. Samples for cross-validation were selected, thereby allowing quantitative evaluation of glucose, fructose and sucrose in all samples. Furthermore, a representative number of honeys were mixed with common adulterant syrups to simulate adulteration procedures and to verify the reliability of the ATR-FTIR technique in recognizing them. Preliminary results revealed not only the quantitative reliability of ATR-FTIR spectroscopy in the evaluation of sugars in honeys, but also its ability in recognizing honey adulteration

From Waste to Resources: Sewage Sludges from the Citrus Processing Industry to Improve Soil Fertility and Performance of Lettuce (Lactuca sativa L.)

The citrus industry produces a large number of sludges as a consequence of citrus wastewater treatment. The correct disposal of citrus sewage sludges (CSSs) has been attempted using anaerobic digestion, aerobic digestion, and lime stabilization. However, since CSSs hold nitrogen, phosphorus, and other macronutrients required by crops, in line with the circular economy principles, they could be utilized for agricultural purposes, such as organic fertilizer. The aim of this study was to assess the effect of CSSs supplied at different doses on soil fertility and lettuce performance. To this end, a pot experiment was established. The soil was amended with CSSs at three different concentrations (2.5, 5, 10 t ha(-1)). After 46 days of lettuce growth, the experiment was stopped, and soils and plants were analyzed. Soil amended with CSSs showed an increase in total organic C ranging from 7% to 11%. Additionally, available P increased but only at the highest CSS dose. The addition of CSSs affected the biochemical properties of soil, but a univocal trend related to the number of CSSs applied was not found. Microbial biomass C increased only with the highest dose of CSS applied, while the metabolic quotient (qCO(2)) decreased. Such a positive effect on soil fertility and soil microorganisms, in turn, lead to an increase in lettuce biomass. Moreover, results indicated that following CSS addition, lettuce crops adsorbed more N in leaves than in roots, whereas P, Ca, Mg, K, and Na showed an opposite pattern and increased more consistently in roots. In conclusion, amendment with CSSs enhances soil fertility by increasing, regardless of CSS dose, total organic C, and, at the highest dose, P availability and microbial biomass C. Such improvement in soil fertility, in turn, increases lettuce biomass production without affecting its quality, i.e., alteration of the (K + Na)/(Ca + Mg) ratio