Search

Abstract Background The new trend demanding for “natural” agri-food products has encouraged the application of more sustainable and eco-friendly farming methods, which limit or avoid the use of synthetic chemicals. This approach is increasing in viticulture, one of the sectors with the highest commercial value since grapes and derived products are largely consumed foodstuffs, with appreciated nutritional and sensory features. In this work, 1H Nuclear Magnetic Resonance spectroscopy (1H NMR) was applied for the metabolic profiling of cv. Italia table grapes samples, from the same origin area, cultivated with different treatments (biodynamic, organic and integrated) and collected in three subsequent vintages. Multivariate statistical analysis was performed on NMR-data with the aim of comprehensively researching the possible influences on metabolites due to the use of diverse agricultural practices. Results Both inter-annual variability (2020, 2021 and 2022 vintages) and different vineyard treatments (biodynamic, organic and integrated) resulted as significant drivers for samples differentiation in the preliminary unsupervised analysis of the (1H NMR spectra derived) metabolic profile data. Nevertheless, supervised data analyses showed that inter-vineyards variability, due to application of diverse farming methods, had a comparable discriminating effect with respect to harvesting years. Ethanol, sugars (as α-/β-glucose), organic acids (as malate) and amino acids (as arginine, leucine, glutamine) resulted the most viticultural practices-dependent metabolites. Interestingly, results from pairwise comparisons between treatments indicated the biodynamic samples with respect to the organic ones as the best-observed differentiation. This was followed by the biodynamic vs integrated and organic vs integrated samples comparisons, in decreasing discrimination order, as confirmed by the descriptiveness and predictive ability parameters of the corresponding pairwise OPLS-DA models. Conclusions Results highlighted that metabolites’ composition in cv. Italia table grapes juice is significantly affected by the use of different kinds of vineyard managements (biodynamic, organic and integrated, here investigated). Metabolomics study, here employing 1H NMR spectroscopy combined with multivariate statistical analysis, offers powerful tools to elucidate the metabolic differences among classes of samples. Graphical Abstract

(1) Background: A preliminary investigation of Protected Designation of Origin (PDO) wines (red and rosé) produced from Negroamaro grapes—a native Salento (Apulia, Southern Italy) vine that is part of the Salice s.no PDO area—was performed in this work. (2) Methods: 1H-NMR spectroscopy, in combination with multivariate statistical analysis (MVA), was employed to characterize the metabolic profiles of 39 wine samples. Spectrophotometric methods were used to obtain preliminary information on the phenolic composition of wines and the associated antioxidant activity. The HepG2 liver cell line was used to assess the biological activity (effect on cell viability and genotoxicity activity) of wine samples. (3) Results: The NMR spectra analysis revealed the presence of signals ascribable to phenolic compounds such as gallic, hydroxycinnamic, and syringic acids. Relative content of these metabolites has been shown to be higher in red than in rosés wines and related to the wine producers. Interestingly, a similar pattern was observed in biological analyses. Red wines compared to the rosé wines display great variations in antioxidant capacity when evaluated as fresh samples using the DPPH and ORAC methods. Furthermore, all red wines exhibited a concentration-dependent decrease in cellular viability and live cells; this phenomenon is much less pronounced in rosé wines. (4) Conclusions: The resulting findings from this study reveal that winemaking operations could lead to final products with different chemical compositions and related properties. Even when starting from the same crop variety and cultivation region, significant differences were observed in the wine samples NMR-metabolic profiles and in vitro biotoxicological activity.

This study was designed as a randomized, placebo-controlled, double-blinded, cross-over trial performed to investigate the effects of a dietary supplement containing undenatured type II collagen (UCII®) and Boswellia Serrata on mobility, pain and joint metabolism in mild moderate osteoarthritis (OA) in dogs. A total of 60 dogs with mobility problems were evaluated and enrolled in the study. Seventeen of these dogs with mild/moderate OA were randomized to receive the product A (UCII® + Boswellia Serrata supplement-UCII®-BW) or product B (Placebo -PL), 1 chew per day for 8 weeks by oral route, and repeated in a crossover design after 4 weeks of washout period. All the subjects had veterinary evaluations during the trial and owners were requested to fill out a questionnaire on mobility impairment using the Liverpool Osteoarthritis in dogs scale (L.O.A.D.) at each time of the study. Objective tools were used to assess mobility, activity, and pain. Metabolomic analysis was performed on synovial fluid of most affected joint at the beginning and the end of the study. The results proved that UCII®+Boswellia serrata supplemented group over a period of eight weeks results in an improvement of mobility impairment, already at 4 weeks of administration, according to the owner´s evaluation. In contrast, its absence increased the risk of OA crisis and decreased the pain threshold on the most affected joint. Furthermore, the synovial fluid metabolic profile showed moderate differences between the beginning and the end of the supplementation period, with a particular influence associated to the time of UCII®-BW administration.

In 2013, an outbreak of Xylella fastidiosa (Xf) was identified for the first time in Europe, in the extreme south of Italy (Apulia, Salento territory). The locally identified subspecies pauca turned out to be lethal for olive trees, starting an unprecedented phytosanitary emergency for one of the most iconic cultivations of the Mediterranean area. Xf pauca (Xfp) is responsible for a severe disease, the olive quick decline syndrome (OQDS), spreading epidemically and with dramatic impact on the agriculture, the landscape, the tourism and the cultural heritage of this region. The bacterium, transmitted by insects that feed on xylem sap, causes rapid wilting in olive trees due to biofilm formation, which obstructs the plant xylematic vessels. The aim of this review is to perform a thorough analysis that offers a general overview of the published work, from 2013 to December 2023, related to the Xfp outbreak in Apulia. This latter hereto has killed millions of olive trees and left a ghostly landscape with more than 8000 square kilometers of infected territory, that is 40% of the region. The majority of the research efforts made to date to combat Xfp in olive plants are listed in the present review, starting with the early attempts to identify the bacterium, the investigations to pinpoint and possibly control the vector, the assessment of specific diagnostic techniques and the pioneered therapeutic approaches. Interestingly, according to the general set criteria for the preliminary examination of the accessible scientific literature related to the Xfp outbreak on Apulian olive trees, fewer than 300 papers can be found over the last decade. Most of them essentially emphasize the importance of developing diagnostic tools that can identify the disease early, even when infected plants are still asymptomatic, in order to reduce the risk of infection for the surrounding plants. On the other hand, in the published work, the diagnostic focus (57%) overwhelmingly encompasses all other possible investigation goals such as vectors, impacts and possible treatments. Notably, between 2013 and 2023, only 6.3% of the literature reports addressing the topic of Xfp in Apulia were concerned with the application of specific treatments against the bacterium. Among them, those reporting field trials on infected plants, including simple pruning indications, were further limited (6%).

The study of plant metabolome and the role of cellular pathway end products has gained increased attention [...]

Abstract The aim of the study was to compare the metabolomic synovial fluid (SF) profile of dogs affected by spontaneous osteoarthritis (OA) and supplemented with undenatured type II collagen (UC-II), with that of healthy control dogs. Client-owned dogs were enrolled in the study and randomized in two different groups, based on the presence/absence of OA (OA group and OA-free group). All dogs were clinically evaluated and underwent SF sampling for 1H-Nuclear Magnetic Resonance spectroscopy (1H-NMR) analysis at time of presentation. All dogs included in OA group were supplemented with UC-II orally administered for 30 days. After this period, they were reassessed (OA-T30). The differences in the 1H-NMR metabolic SFs profiles between groups (OA-free, OA-T0 and OA-T30) were studied. The multivariate statistical analysis performed on SFs under different conditions (OA-T0 vs OA-T30 SFs; OA-T0 vs OA-free SFs and OA-T30 vs OA-free SFs) gave models with excellent goodness of fit and predictive parameters, revealed by a marked separation between groups. β-Hydroxybutyrate was identified as a characteristic compound of osteoarthritic joints, showing the important role of fat metabolism during OA. The absence of β-hydroxybutyrate after UC-II supplementation suggests the supplement’s effectiveness in rebalancing the metabolism inside the joint. The unexpectedly high level of lactate in the OA-free group suggests that lactate could not be considered a good marker for OA. These results prove that 1H-NMR-based metabolomic analysis is a valid tool to study and monitor OA and that UC-II improves clinical symptoms and the SF metabolic profile in OA dogs.

Pomegranate (Punica granatum L.) fruits are a historical agricultural product of the Mediterranean basin that became increasingly popular in the latest years for being rich in antioxidants and other micronutrients, and are extensively commercialized as fruits, juice, jams and, in some Eastern countries, as a fermented alcoholic beverage. In this work, four different pomegranate wines specifically designed using combinations of two cultivars (Jolly Red and Smith) and two yeast starters with markedly different characteristics (Saccharomyces cerevisiae Clos and Saccharomyces cerevisiae ex-bayanus EC1118) were analyzed. The chemical characterization of the wines together with the originating unfermented juices was performed by 1H NMR spectroscopy metabolomic analysis. The full spectra were used for unsupervised and supervised statistical multivariate analysis (MVA), namely Principal Component Analysis (PCA), Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA), and sparse PCA (SPCA). The MVA of the wines showed a clear discrimination between the cultivars, and a smaller, yet significant, discrimination between the yeasts used. In particular, a higher content of citrate and gallate was observed for the Smith cv. and, on the contrary, a statistically significant higher content of fructose, malate, glycerol, 2,3 butanediol, trigonelline, aromatic amino acids and 4-hydrophenylacetate was observed in Jolly Red pomegranate wines samples. Significant interaction among the pomegranate cultivar and the fermenting yeast was also observed. Sensorial analysis was performed by a panel of testing experts. MVA of tasting data showed that the cultivar significantly affected the organoleptic parameters considered, while the yeast had a minor impact. Correlation analysis between NMR-detected metabolites and organoleptic descriptors identified several potential sensorially-active molecules as those significantly impacting the characteristics of the pomegranate wines.

Abstract Xylella fastidiosa is a xylem-limited bacterium causing a range of economically important plant diseases in hundreds of crops. Over the last decade, a severe threat due to Olive Quick Decline Syndrome (OQDS), caused by Xylella fastidiosa subspecies pauca, affected the Salento olive groves (Apulia, South-East Italy). Very few phyto-therapeutics, including a Zn/Cu citric acid biocomplex foliar treatment, were evaluated to mitigate this disease. However, the traditional foliar applications result in the agro-actives reaching only partially their target. Therefore the development of novel endo-therapeutic systems was suggested. Metabolite fingerprinting is a powerful method for monitoring both, disease progression and treatment effects on the plant metabolism, allowing biomarkers detection. We performed, for the first time, short-term monitoring of metabolic pathways reprogramming for infected Ogliarola salentina and Cima di Melfi olive trees after precision intravascular biocomplex delivery using a novel injection system. Upon endo therapy, we observed specific variations in the leaf content of some metabolites. In particular, the 1H NMR-based metabolomics approach showed, after the injection, a significant decrease of both the disease biomarker quinic acid and mannitol with simultaneous increase of polyphenols and oleuropein related compounds in the leaf’s extracts. This combined metabolomics/endo-therapeutic methodology provided useful information in the comprehension of plant physiology for future applications in OQDS control.

Metabolomics is a powerful tool in food sciences, widely used in food analysis for authenticity and traceability assessment and regulatory compliance, processing, quality, and safety [...]

Here we report the medium-term effects of foliar spray and endo-therapy treatments with different doses of a Cu/Zn citric acid biocomplex (Dentamet®) in Xylella fastidiosa infected olive trees of Salento, Apulia region (South-east Italy). Leaf extract samples from field-treated 150 years old olive trees cvs Ogliarola salentina and Cellina di Nardò were studied by 1H NMR-based metabolomics. The result of different applications of Dentamet® endo-therapy after 60, 120 and 180 days in comparison with traditional foliar spray treatment and water injection as a control have been investigated. The metabolic profile analyses, performed by 1H NMR-based metabolomic approach, indicated plant metabolites variations connected to the disease progression such as mannitol, quinic acid, and oleuropein related compounds. The best results, in terms of discrimination of the metabolic profiles with respect to water injection, were found for monthly endo-therapy treatments. Dentamet® foliar application demonstrated more specific time related progressive effectiveness with respect to intravascular treatments. Therefore, besides a possible more effective performance of endo-therapy with respect to foliar treatments, the need of further doses/frequencies trimming to obtain long-term results was also assessed. The present field studies confirmed the indication of Dentamet® effectiveness in metabolic variation induction, potentially linked with reducing the X. fastidiosa subspecies pauca related Olive Quick Decline Syndrome (OQDS) symptoms development.

Abstract Hepatocellular carcinoma (HCC) is nowadays the sixth cause of tumour-related deceases worldwide, estimated to become the third in Western countries by 2030. New drugs for HCC treatment still have many adverse effects. Several lines of evidence indicate that plant metabolites offer concrete opportunities for developing new therapeutic strategies for many diseases, including cancer. We previously reported that ethyl acetate extract of a spontaneous edible plant harvested in Apulia, Crithmum maritimum, significantly inhibited cell growth in HCC cells. By 1H-NMR spectroscopy, here we show that Crithmum maritimum ethyl acetate extract counteracts the Warburg effect, by reducing intracellular lactate, inhibits protein anabolism, by decreasing amino acid level, and affects membrane biosynthesis by lowering choline and phosphocholine. Also, we observed an effect on lipid homeostasis, with a reduction in triglycerides, cholesterol, monounsaturated fatty acids (MUFA), and diunsaturated fatty acids (DUFA), and an increase in polyunsaturated fatty acids (PUFA). Taken together, these data demonstrate that Crithmum maritimum-induced cytostasis is exerted through a multi-effect action, targeting key metabolic processes in HCC cells. Overall, our findings highlight the role of Crithmum maritimum as a promising tool for the prevention and the improvement of the therapeutic options for HCC and other types of tumours.

This study aims to evaluate the Proton Nuclear Magnetic Resonance (1H-NMR)-based metabolomic profiles of monovarietal olive oils obtained from Arbequina and Koroneiki, which are typically present in the Mediterranean basin and grown in Spain and Greece, respectively. Micro-milled oils were obtained from olives harvested both in their cultivar home countries (Spain and Greece) and in Italy. Investigations by 1H NMR metabolic profiling were carried out to develop a strategy for the correct interpretation of differences based on geographical origins. The NMR Spectroscopy and multivariate statistical analysis (MVA) revealed significant differences in fatty acids profile as well as the unsaponifiable fraction, not only according to the cultivars but also to the specific geographical origin of the olives used. In particular, the oils from Spain were higher in polyunsaturated fatty acids (PUFAs) content than those from Italy. Conversely, the Italian oils of Koroneiki showed higher content of monounsaturated fatty acids (MUFAs) than Greek oils. Regarding the Extra Virgin Olive Oils (EVOOs) minor fraction, for both cultivars, the olive oils obtained using olives harvested in Italy were characterized by higher relative content of phenolic compounds. It was found that each of the investigated cultivars (Arbequina and Koroneiki), very popular in super high density (SHD) planting systems, assumed a specific well-characterized metabolic EVOO profile when the olives are harvested in Italy. These results may contribute to extending and enforcing available literature data on 1H NMR-based chemometric models as powerful tools for EVOOs geographical origin discrimination.

Improved cellulose biosynthesis and plant biomass represent important economic targets for several biotechnological applications including bioenergy and biofuel production. The attempts to increase the biosynthesis of cellulose by overexpressing CesAs proteins, components of the cellulose synthase complex, has not always produced consistent results. Analyses of morphological and molecular data and of the chemical composition of cell walls showed that tobacco plants (F31 line), stably expressing the Arabidopsis CesA6 fused to GFP, exhibits a “giant” phenotype with no apparent other morphological aberrations. In the F31 line, all evaluated growth parameters, such as stem and root length, leaf size, and lignified secondary xylem, were significantly higher than in wt. Furthermore, F31 line exhibited increased flower and seed number, and an advance of about 20 days in the anthesis. In the leaves of F31 seedlings, the expression of primary CesAs (NtCesA1, NtCesA3, and NtCesA6) was enhanced, as well as of proteins involved in the biosynthesis of non-cellulosic polysaccharides (xyloglucans and galacturonans, NtXyl4, NtGal10), cell wall remodeling (NtExp11 and XTHs), and cell expansion (NtPIP1.1 and NtPIP2.7). While in leaves the expression level of all secondary cell wall CesAs (NtCesA4, NtCesA7, and NtCesA8) did not change significantly, both primary and secondary CesAs were differentially expressed in the stem. The amount of cellulose and matrix polysaccharides significantly increased in the F31 seedlings with no differences in pectin and hemicellulose glycosyl composition. Our results highlight the potentiality to overexpress primary CesAs in tobacco plants to enhance cellulose synthesis and biomass production.

Geographical origin assessment of extra virgin olive oil (EVOO) is recognised worldwide as raising consumers’ awareness of product authenticity and the need to protect top-quality products. The need for geographical origin assessment is also related to mandatory legislation and/or the obligations of true labelling in some countries. Nevertheless, official methods for such specific authentication of EVOOs are still missing. Among the analytical techniques useful for certification of geographical origin, nuclear magnetic resonance (NMR) and mass spectroscopy (MS), combined with chemometrics, have been widely used. This review considers published works describing the use of these analytical methods, supported by statistical protocols such as multivariate analysis (MVA), for EVOO origin assessment. The research has shown that some specific countries, generally corresponding to the main worldwide producers, are more interested than others in origin assessment and certification. Some specific producers such as Italian EVOO producers may have been focused on this area because of consumers’ interest and/or intrinsic economical value, as testified also by the national concern on the topic. Both NMR- and MS-based approaches represent a mature field where a general validation method for EVOOs geographic origin assessment could be established as a reference recognised procedure.

Abstract Background Xylella fastidiosa is a Gram-negative bacterium which lives in the xylem of plants, causing its occlusion and other alterations inducing eventually the death of the infected plants. In Salento, the sub-peninsula in the south-eastern of Apulia Region (southern Italy), the infection of X. fastidiosa has been associated with the widespread presence of CoDiRO (complex of parasitic agents that constitute the so-called “olive quick decline syndrome”) and currently represents a serious local emergence. The need to adopt specific agronomic measures to contrast the further disease spread has been recently raised. The extensive NMR-based metabolomic approach to study the metabolic effects of CoDiRO on local olive cultivars such as Ogliarola salentina and Cellina di Nardò was used. Results In this study, the effects of a CE approved fertilizer containing zinc, copper, and citric acid, known as DENTAMET®, on CoDiRO-exhibiting olive trees infected by X. fastidiosa were studied by 1H NMR spectroscopy. The changes in the metabolomic profiles of aqueous extracts obtained from leaves of the two olive cultivars are reported. Upon the DENTAMET® treatments, different and opposite polyphenolic and sugars patterns in the two cultivars, which showed a different incidence and severity of disease before the treatments, were detected. Conclusions Differences in the sugars and polyphenols content of treated versus untreated trees could potentially contribute to the syndrome monitoring and might be related to the X. fastidiosa presence. Graphical abstract Apulian (Salento) CoDiRO-exhibiting olive trees infected by X. fastidiosa have been studied by 1H NMR spectroscopy. Changes in the metabolomic profiles are reflected in different sugars and polyphenols content after DENTAMET®-treatment

Xylella fastidiosa subsp. pauca is associated with the olive quick decline syndrome in Salento (Apulia region, Southern Italy). The first outbreak of the disease was noticed in the Gallipoli district, from where it subsequently reached nearby areas. To date, no specific study has verified if abnormal soil or leaf micronutrients is associated with the disease. Soil and leaf samples were taken from 23 olive farms showing symptoms of the syndrome located in Gallipoli and nearby areas. Each sample was analyzed for magnesium and micronutrients content using inductively coupled plasma atomic emission spectroscopy. Real-time PCR indicated that X. f. subsp. pauca was present in each sampled tree. There is a general lack of molybdenum in soil and low bioavailability of copper and molybdenum in tree leaves. Low content of manganese in soil was also found in some farms located in Gallipoli, Galatone and Trepuzzi. Olive trees grown in Gallipoli area also had low content of boron. Principal component analysis showed that soil and leaf samples from this area had lower micronutrient contents compared with other areas. General copper depletion in leaves was uncommon and has not been previously recorded in Italy in a large area. This could indicate that X. f. subsp. pauca infection causes a depletion of copper within olive leaves. Reduced copper content has been previously recorded in leaves infected with X. f. subsp. multiplex and for other bacterial pathogens. The role of copper in relation to X. fastidiosa infection is discussed.

Xylella fastidiosa subsp. pauca is the causal agent of “olive quick decline syndrome” in Salento (Apulia, Italy). On April 2015, we started interdisciplinary studies to provide a sustainable control strategy for this pathogen that threatens the multi-millennial olive agroecosystem of Salento. Confocal laser scanning microscopy and fluorescence quantification showed that a zinc-copper-citric acid biocomplex—Dentamet®—reached the olive xylem tissue either after the spraying of the canopy or injection into the trunk, demonstrating its effective systemicity. The biocomplex showed in vitro bactericidal activity towards all X. fastidiosa subspecies. A mid-term evaluation of the control strategy performed in some olive groves of Salento indicated that this biocomplex significantly reduced both the symptoms and X. f. subsp. pauca cell concentration within the leaves of the local cultivars Ogliarola salentina and Cellina di Nardò. The treated trees started again to yield. A 1H-NMR metabolomic approach revealed, upon the treatments, a consistent increase in malic acid and γ-aminobutyrate for Ogliarola salentina and Cellina di Nardò trees, respectively. A novel endotherapy technique allowed injection of Dentamet® at low pressure directly into the vascular system of the tree and is currently under study for the promotion of resprouting in severely attacked trees. There are currently more than 700 ha of olive groves in Salento where this strategy is being applied to control X. f. subsp. pauca. These results collectively demonstrate an efficient, simple, low-cost, and environmentally sustainable strategy to control this pathogen in Salento.

Considering the growing number of extra virgin olive oil (EVOO) producers in the world, knowing the influence of olive oils with different geographical origins on the characteristics of the final blend becomes an interesting goal. The present work is focused on commercial organic EVOO blends obtained by mixing multiple oils from different geographical origins. These blends have been studied by 1H-NMR spectroscopy supported by multivariate statistical analysis. Specific characteristics of commercial organic EVOO blends originated by mixing oils from Italy, Tunisia, Portugal, Spain, and Greece were found to be associated with the increasing content of the Italian component. A linear progression of the metabolic profile defined characteristics for the analysed samples—up to a plateau level—was found in relation to the content of the main constituent of the Italian oil, the monocultivar Coratina. The Italian constituent percentage appears to be correlated with the fatty acids (oleic) and the polyphenols (tyrosol, hydroxytyrosol, and derivatives) content as major and minor components respectively. These results, which highlight important economic aspects, also show the utility of 1H-NMR associated with chemometric analysis as a powerful tool in this field. Mixing oils of different national origins, to obtain blends with specific characteristics, could be profitably controlled by this methodology.

Xylella fastidiosa subsp. pauca, is a bacterial phytopathogen associated with the “olive quick decline syndrome” (OQDS) causing severe economic losses to olive groves in Salento area (Apulia, Italy). In a previous work, we analyzed by 1H-NMR the metabolic pattern of naturally infected Ogliarola salentina and Cellina di Nardò susceptible cultivars untreated and treated with a zinc-copper citric acid biocomplex and we observed the treatment related variation of the disease biomarker quinic acid. In this study, we focused also on the Leccino cultivar, known to exhibit tolerance to the disease progression. The 1H-NMR-based metabolomic approach was applied with the aim to characterize the overall metabolism of tolerant Leccino in comparison with the susceptible cultivars Ogliarola salentina and Cellina di Nardò under periodic mid-term treatment. In particular, we studied the leaf extract molecular patterns of naturally infected trees untreated and treated with the biocomplex. The metabolic Leccino profiles were analyzed for the first time and compared with those exhibited by the susceptible Cellina di Nardò and Ogliarola salentina cultivars. The study highlighted a specificity in the metabolic response of the tolerant Leccino compared to susceptible cultivars. These differences provide useful information to describe the defensive mechanisms underlying the change of metabolites as a response to the infection, and the occurrence of different levels of disease, season and treatment effects for olive cultivars.

During the last few years, the global demand for extra virgin olive oil (EVOO) is increased. Olive oil represents a significant percentage of world fat consumption determining an important development of its market. In this context, the problems related to counterfeiting and product fraud is becoming extremely relevant. Thus, the quality and authenticity control of EVOOs is nowadays mandatory. In this study we focused on the use of 1H NMR technique associated with multivariate statistical analysis to characterize Italian EVOOs commercial blends. In particular, a specific database including 126 monocultivar EVOOs reference samples, was used to characterize a total of 241 Italian EVOOs blends over four consecutive harvesting years. Moreover, the effect of the minor components (phenolic compounds) on the qualitative characterization of blended EVOOs was also evaluated. The correlation analysis of classification scores obtained using two pairwise orthogonal partial least square-discriminant analysis models (built with major and combined major–minor components NMR data) revealed that both could be profitably used to generally classify the studied Coratina containing blends.

Xylella fastidiosa subsp. pauca is responsible for the “olive quick decline syndrome” (OQDS) in Salento (Apulia). The main epidemiological aspects of the syndrome are related to the pathogen spread and survival in the area, and to the biology of the insect vector. The assessment of the macro and microelements content (i.e., ionome) in soil and leaves could provide basic and useful information. Indeed, knowledge of host ionomic composition and the possibility of its modification could represent a potential tool for the management of diseases caused by X. fastidiosa. Therefore, soil and leaf ionomes of naturally infected, not infected, and zinc–copper–citric acid biocomplex treated trees of different areas of Apulia and the bordering Basilicata regions were compared. We observed that soil and leaf ionomic composition of olive farms growing in the pathogen-free areas north of the Salento Barletta-Andria-Trani BAT (Apulia) and Potenza PZ (Basilicata, Apulia bordering region) provinces is significantly different from that shown by the infected olive groves of the Salento areas (LE, BR, TA provinces). In particular, a higher content of zinc and copper both in soil and leaves was found in the studied northern areas in comparison to the southern areas. This finding could partly explain the absence of OQDS in those areas. In the infected Salento areas, the leaf ionomic profile resulted as being markedly different for the biocomplex treated compared to the untreated trees. A higher zinc content in leaves characterized treated with respect to untreated trees. On the other hand, among the not-infected trees, Xylella-resistant Leccino showed higher manganese content when compared with the higher pathogen sensitive Ogliarola salentina and Cellina di Nardò. According to these results, soil and olive leaf ionome could provide basic information for the epidemiologic study and possible control of X. f. subsp. pauca in Apulia.

The Mediterranean basin is one of the regions heavily affected by jellyfish bloom phenomena, mainly due to the presence of scyphozoans, such as Rhizostoma pulmo. The jellyfish have few natural predators, and their bodies represent an organic-rich substrate that can support rapid bacterial growth with great impact on the structure of marine food webs. In Asiatic countries, jellyfish are widely studied for their health benefits, but their nutritional and nutraceutical values still remain poorly characterized. In this study, the differences in the 1H NMR spectroscopy metabolic profiles of R. pulmo female gonads and body fractions (including umbrella and oral arms), in different sampling periods, were studied. For each body compartment both lipid and aqueous extracts were characterized and their 1H NMR metabolic profiles subjected to multivariate analysis. From a statistical analysis of the extracts, a higher contents of ω-3 polyunsaturated fatty acids (PUFAs), amino acid and osmolytes (homarine, betaine, taurine) with important roles in marine invertebrates were observed in female gonads, whereas umbrella and oral arms showed similar metabolic profiles. These results support a sustainable exploitation of the jellyfish for the extraction of bioactive compounds useful in nutraceutical, nutricosmetics, and functional food fields.

The bacterium Xylella fastidiosa subsp. pauca is associated with the “olive quick decline syndrome” in the Apulia region of southern Italy. To investigate control of this phytopathogen, a compound containing zinc and copper complexed with citric-acid hydracids (Dentamet®) was evaluated for in vitro and in planta bactericidal activity. Confocal laser scanning microscopy, fluorescent quantification and atomic emission spectroscopy were then used to determine if the compound reached the xylem networks of leaves, twigs and branches of olive, to release zinc and copper within the xylem. A 3-year field trial in an olive orchard containing mature Cellina di Nardò and Ogliarola salentina olive trees, and officially declared infected by X. fastidiosa subsp. pauca,was also carried out o to determine if the compound affected severity of the disease. Each year, from early April to October (excluding July and August), six spray treatments of 0.5% (v:v) Dentamet® were applied on the olive tree crowns. The compound reduced severity of symptoms in both cultivars. Most untreated trees died by the end of the trial, whereas all treated trees survived with good vegetative status as assessed by a normalized difference vegetation index. Quantitative real-time PCR was performed from June 2016 to September 2017, following the official procedures established by the European and Mediterranean Plant Protection Organization. The analysis revealed a statistically significant reduction of X. fastidiosa cell densities within the leaves of treated trees. These promising results suggest that integrated management to reduce severity of X. fastidiosa that includes regular pruning and soil harrowing with spring and summer spray treatments with Dentamet®, is likely to effectively control the disease.

Xylella fastidiosa subsp. pauca is a xylem-limited bacterial phytopathogen currently found associated on many hectares with the “olive quick decline syndrome” in the Apulia region (Southern Italy), and the cultivars Ogliarola salentina and Cellina di Nardò result in being particularly sensitive to the disease. In order to find compounds showing the capability of reducing the population cell density of the pathogen within the leaves, we tested, in some olive orchards naturally-infected by the bacterium, a zinc-copper-citric acid biocomplex, namely Dentamet®, by spraying it to the crown, once per month, during spring and summer. The occurrence of the pathogen in the four olive orchards chosen for the trial was molecularly assessed. A 1H NMR metabolomic approach, in conjunction with a multivariate statistical analysis, was applied to investigate the metabolic pattern of both infected and treated adult olive cultivars, Ogliarola salentina and Cellina di Nardò trees, in two sampling periods, performed during the first year of the trial. For both cultivars and sampling periods, the orthogonal partial least squares discriminant analysis (OPLS-DA) gave good models of separation according to the treatment application. In both cultivars, some metabolites such as quinic acid, the aldehydic form of oleoeuropein, ligstroside and phenolic compounds, were consistently found as discriminative for the untreated olive trees in comparison with the Dentamet®-treated trees. Quinic acid, a precursor of lignin, was confirmed as a disease biomarker for the olive trees infected by X. fastidiosa subsp. pauca. When treated with Dentamet®, the two cultivars showed a distinct response. A consistent increase in malic acid was observed for the Ogliarola salentina trees, whereas in the Cellina di Nardò trees the treatments attenuate the metabolic response to the infection. To note that in Cellina di Nardò trees at the first sampling, an increase in γ-aminobutyric acid (GABA) was observed. This study highlights how the infection incited by X. fastidiosa subsp. pauca strongly modifies the overall metabolism of olive trees, and how a zinc-copper-citric acid biocomplex can induce an early re-programming of the metabolic pathways in the infected trees.

Jellyfish outbreaks in marine coastal areas represent an emergent problem worldwide, with negative consequences on human activities and ecosystem functioning. However, potential positive effects of jellyfish biomass proliferation may be envisaged as a natural source of bioactive compounds of pharmaceutical interest. We investigated the biochemical composition of mature female gonads and lysozyme antibacterial activity of oocytes in the Mediterranean barrel jellyfish Rhizostoma pulmo. Chemical characterization was performed by means of multinuclear and multidimensional NMR spectroscopy. The ovaries of R. pulmo were mainly composed of water (93.7 ± 1.9% of wet weight), with organic matter (OM) and dry weight made respectively of proteins (761.76 ± 25.11 µg mg−1 and 45.7 ± 1.5%), lipids (192.17 ± 10.56 µg mg−1 and 9.6 ± 0.6%), and carbohydrates (59.66 ± 2.72 µg mg−1 and 3.7 ± 0.3%). The aqueous extract of R. pulmo gonads contained free amino acids, organic acids, and derivatives; the lipid extract was composed of triglycerides (TG), polyunsaturated fatty acids (PUFAs), diunsaturated fatty acids (DUFAs), monounsaturated fatty acids (MUFAs), saturated fatty acids (SFAs), and minor components such as sterols and phospholipids. The R. pulmo oocyte lysate exhibited an antibacterial lysozyme-like activity (mean diameter of lysis of 9.33 ± 0.32 mm corresponding to 1.21 mg/mL of hen egg-white lysozyme). The occurrence of defense molecules is a crucial mechanism to grant healthy development of mature eggs and fertilized embryos (and the reproductive success of the species) by preventing marine bacterial overgrowth. As a corollary, these results call for future investigations for an exploitation of R. pulmo biomasses as a resource of bioactive metabolites of biotechnological importance including pharmaceuticals and nutrition.

The biological invasion of the green algae Caulerpa cylindracea represents a serious scientific and public issue in the Mediterranean Sea, essentially due to strong modifications both to habitat structure and native benthic communities. Although alterations in health status and changes in flesh quality of some marine species (dietary exposed to C. cylindracea) have been observed, no studies on cause-effect relationships have been carried out. Here, for the first time, through a controlled feeding experiment followed by 1H NMR Spectroscopy and multivariate analysis (PCA, OPLS-DA), we showed that caulerpin taken with diet is directly responsible of changes observed in metabolic profile of fish flesh, including alteration of lipid metabolism, in particular with a reduction of ω3 PUFA content. The potential of caulerpin to directly modulate lipid metabolism opens up new questions about causal mechanism triggered by algal metabolite also in view of a possible exploitation in the nutraceutical/medical field.

According to Coldiretti, Italy still continues to hold the European Quality record in extra virgin olive oils with origin designation and protected geographical indication (PDO and PGI). To date, 46 Italian brands are recognized by the European Union: 42 PDO and 4 PGI (Tuscan PGI, Calabria PGI; Tuscia PGI and PGI Sicily). Specific regulations, introduced for these quality marks, include the designation of both the geographical areas and the plant varieties contributing to the composition of the olive oil. However, the PDO and PGI assessment procedures are currently based essentially on farmer declarations. Tuscan PGI extra virgin olive oil is one of the best known Italian trademarks around the world. Tuscan PGI varietal platform is rather wide including 31 specific olive cultivars which should account for at least 95% of the product. On the other hand, while the characteristics of other popular Italian extra virgin olive oils (EVOOs) cultivars from specific geographical areas have been extensively studied (such as those of Coratina based blends from Apulia), little is still known about Tuscan PGI EVOO constituents. In this work, we performed, for the first time, a large-scale analysis of Tuscan PGI monocultivar olive oils by 1H NMR spectroscopy and multivariate statistical analyses (MVA). After genetic characterization of 217 leaf samples from 24 selected geographical areas, distributed all over the Tuscany, a number of 202 micro-milled oil samples including 10 PGI cultivars, was studied. The results of the present work confirmed the need of monocultivar genetically certified EVOO samples for the construction of 1H-NMR-metabolic profiles databases suitable for cultivar and/or geographical origin assessment. Such specific PGI EVOOs databases could be profitably used to justify the high added value of the product and the sustainability of the related supply chain.