Influence of flash heating and aspergillopepsin I supplementation on must and wine attributes of aromatic varieties.

The protein instability with haze formation represents one of the main faults occurring in white and rosé wines. Among the various solutions industrially proposed, aspergillopepsin I (AP-I) supplementation coupled with must heating (60-75 °C) has been recently approved by OIV and the European Commission for ensuring protein stability of wines. This study investigates the impact of AP-I either applied independently or in combination with flash pasteurization on the chemical composition of grape must and wines derived from Sauvignon Blanc and Gewürztraminer. The efficacy on protein stability of a complete treatment combining heat (70 °C) and AP-I (HP) was confirmed through heat test and bentonite requirement, although no differences were observed between must heating and HP treatments. However, high-performance liquid chromatography analysis of unstable pathogenesis-related proteins revealed that AP-I supplementation reduced chitinases and thaumatin-like proteins compared to the non-enzymed samples, with and without must heating. Amino acid increase was reported only in HP musts, particularly in Sauvignon Blanc. The concentration of yeast-derived aroma compounds in Gewürztraminer wines was increased by must heating; compared to controls, flash pasteurization rose the overall acetate esters content of 85 % and HP of 43 %, mostly due to isoamyl acetate. However, heat treatments -with or without AP-I- reduced terpenes up to 68 %. Despite the different aroma profiles, no differences were observed for any descriptor for both varieties in wine tasting, and only a slight decrease trend was observed for the floral intensity and the typicality descriptors in heated wines.

Evidence of the Relationship between the Oxygen Radical Absorbance Capacity (ORAC) and the Potential Exposure of White Wines to Atypical Aging (ATA).

Atypical aging (ATA) is a well-known wine defect that leads to unacceptable off-odors of wet mop, soap, waxy, furniture varnish, and dish rag. The main compound that is responsible for these undesired scents, 2-aminoacetophenone (AAP), results from the oxidative degradation of 3-indole acetic acid. This study aimed to investigate different technological factors for a possible effect on the oxygen radical absorption capacity (ORAC) and ATA development in white wine. Oxygenation during fermentation did not result in a significant effect on the development of ORAC or ATA development. Instead, the addition of ascorbic acid after vinification led to increased ORAC and decreased AAP formation. The first evidence of a tendency toward a correlation between ORAC and potential ATA development was found, with r = 0.365. This study showed the potential of the ORAC assay for an early assessment of the possible formation of ATA during wine storage.

Evaluating the Atypical Aging Potential Development in Sparkling Wines Can Be Achieved by Assessing the Base Wines at the End of the Alcoholic Fermentation.

Traditional sparkling wine production is a lengthy and costly process, involving a double fermentation step and a period of aging sur lie; thus, monitoring quality during the key manufacturing stages is crucial. The effects of the second fermentation on the development of 2-aminoacetophenone (AAP), the main marker of the atypical aging (ATA) defect, were investigated on 55 base wines (BWs) and corresponding sparkling wines (SWs) produced in an experimental winery. While the AAP content of the SWs was observed to be higher than the BWs, it was found that an artificial aging test carried out on the BWs could be a good predictor of ATA development in SWs. Further, the antioxidant capacity of the SWs was noticed to correlate well with the potential AAP formed during accelerated aging. Finally, an analysis of covariance (ANCOVA) model of linearization capable of predicting AAP formation in SWs using the data obtained from the corresponding BWs was created.

Monitoring hydrogen sulfide de-novo formation from polysulfides present in wine using ion chromatography and ultra high-pressure liquid chromatography combined with fraction collection and high-resolution mass spectrometry.

The study of polysulfides has been a recent topic of interest for wine research due to the possibility of these compounds to release hydrogen sulfide (H2S) during storage. However, studying these compounds has been challenging for several reasons. Polysulfides are low in concentration in natural samples, they are chemically unstable and pure standards of the single compounds (RSnR with n > 2) are not commercially available. In the present study, a method was developed in order to collect a single polysulfide and study its degradation and the consequent formation of H2S. For this approach, ultra-high pressure liquid chromatography was used with an integrated fraction collector and subsequently coupled to high resolution mass spectrometry. After fractionation, the degradation of the di-cysteinyl pentasulfide (CS5C) was induced by exposure to 30 °C and the H2S formation was measured in parallel using ion-exchange chromatography. This method showed the evolutions of different polysulfides and the H2S release originating from the target compound, an observation that to the best of our knowledge has never been made before. The method in the present study demonstrated promising applications for polysulfide studies and brought us a step closer to the understanding of the chemistry of polysulfides in wine.

Atypical ageing defect in Pinot Blanc wines: a comparison between organic and conventional production management systems.

BACKGROUND: Atypical ageing (ATA) is an aroma defect that occurs in white wines and entails a loss of varietal aromas as well as scents of wet mop, shoe polish and dish rag. 2-Aminoacetophenone (2AAP) - a degradation product of indole-3-acetic acid (IAA) - has been described as the main odour-active compound and chemical marker responsible for this off-flavour. A stress reaction in the vineyard triggered by climatic, pedological and viticultural factors can ultimately cause ATA development in wines and remarkably affect wine quality. The aim of this research was to investigate the influence of three grapevine management systems on the occurrence of ATA. The experiments were carried out on Pinot Blanc grape samples from vines cultivated using one conventional and two organic approaches. The management systems mainly differed for the fertilisation regime and the weed control. RESULTS: The amino acid profiles as well as 2AAP and its precursors were quantified in musts and wines using ultra-high-performance liquid chromatography coupled to a high-resolution mass spectrometer. The results showed the existence of a strong vintage effect, while no influence of the use of different agronomic systems was observed. CONCLUSION: The study revealed that an efficient implementation of different grapevine production systems did not affect ATA development in Pinot Blanc wines. This finding is of great relevance for winegrowers and winemakers as it demonstrates that a well-planned organic management system correctly adjusted to the climatic conditions does not pose a threat towards the development of ATA-related compounds in wine. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The anti-snake activity of Nectandra angustifolia flavonoids on phospholipase A2: In vitro and in silico evaluation.

ETHNOPHARMACOLOGICAL RELEVANCE: Lauraceae family includes Nectandra angustifolia a species widely used in the folk medicine of South America against various maladies. It is commonly used to treat different types of processes like inflammation, pain, and snakebites. Snakes of the Bothrops genus are responsible for about 97% of the ophidic accidents in northeastern Argentina. AIM OF THE STUDY: To evaluate the anti-snake activity of the phytochemicals present in N. angustifolia extracts, identify the compounds, and evaluate their inhibitory effect on phospholipase A2 (PLA2) with in vitro and in silico assays. METHODS: Seasonal variations in the alexiteric potential of aqueous, ethanolic and hexanic extracts were evaluated by inhibition of coagulant, haemolytic, and cytotoxic effects of B. diporus venom. The chemical identity of an enriched fraction obtained by bio-guided fractioning was established by UPLC-MS/MS analysis. Molecular docking studies were carried out to investigate the binding mechanisms of the identified compounds to PLA2 enzyme from snake venom. RESULTS: All the extracts inhibited venom coagulant activity. However, spring ethanolic extract achieved 100% inhibition of haemolytic activity. Bio-guide fractioning led to an enriched fraction (F4) with the highest haemolytic inhibition. Five flavonoids were identified in this fraction; molecular docking and Molecular Dynamics (MD) simulations indicated the binding mechanisms of the identified compounds. The carbohydrates present in some of the compounds had a critical effect on the interaction with PLA2. CONCLUSION: This study shows, for the first time, which compounds are responsible for the anti-snake activity in Nectandra angustifolia based on in vitro and in silico assays. The results obtained in this work support the traditional use of this species as anti-snake in folk medicine.

Alkaloid profiling of Italian alpine herbs using high resolution mass spectrometry (Orbitrap-MS).

Alpine pastures have not yet been extensively studied with regard to the presence of alkaloids in herbaceous plants. In this work the alkaloid profiles were characterised from a selection of 62 herbs collected from alpine pastures in north-eastern Italy. High-performance liquid chromatography coupled to a hybrid quadrupole-orbitrap mass spectrometer was used to evaluate the presence of 41 different alkaloids and quantify them using a targeted approach. Provisionally, 118 alkaloids were identified, including both free and glycosylated forms, making use of a homemade database and a suspect screening approach. rucifoline, gramine, heliotrine, lycopsamine, seneciphylline, and veratramine were quantified with concentrations ranging from 6 to about 100 µg kg-1 in 6 plants. Herbaceous plants belonging to the most well-represented plant families (Poaceae, 9 species; Asteraceae, 7; Lamiaceae, 6) showed distinct and characteristic alkaloid profiles and were correctly reclassified with an average accuracy of 85% (Partial Least Squares - Discriminant Analysis).

Hydrochar and hydrochar co-compost from OFMSW digestate for soil application: 2. agro-environmental properties.

The work concerns the study of the hydrochar from digestate and hydrochar co-compost characterization as amendments. The processes for hydrochar and co-compost production were described in Part 1 of this work (Scrinzi et al., 2022). The amendment properties of hydrochar (produced at 180-200-220 °C for 3 h) and co-composts (25%, 50%, and 75% hydrochar percentage of digestate substitution) were assessed by phytotoxicity, plant growth bioassay, and soil effect. Different seeds species (Lepidium sativum, Cucumis sativus, and Sorghum bicolor sp.) were dosed at increased concentrations using both wet raw amendments and their water extracts. The chemical characterization showed phytotoxic compounds content depending on both the initial feedstock (digestate) and the HTC process; at the same time, the analysis highlighted the reduction of these compounds by composting (organic acid, polyphenols, salt concentration). The dose-response was analyzed by the Cedergreen-Streibig-Ritz model and the half-maximal effective concentration (EC50) was calculated based on this equation. The soil properties and GHG emissions measurements (CH4, CO2, N2O, and NH3) highlighted the effect on N dynamics and on soil respiration induced by substrates. The HC200 soil application determined a significant impact on CO2 and N2O emission and NH3 volatilization (10.82 mol CO2/m2; 51.45 mmol N2O/m2; 112 mol NH3/m2) and a significant reduction of total N and TOC (46% of TKN and 49% of TOC). The co-compost (75%) showed specific effects after soil application compared to other samples an increase of available P (48%), a greater content of nitrogen (1626 mg/kg dry basis), and a reduction of organic carbon (17%). Our results demonstrate the good quality of co-compost and at the same time the validity of this post-treatment for addressing many issues related to hydrochar use in the soil as an amendment, confirming the suitability of HTC process integration for digestate treatment in anaerobic digestion plants.

Tannat Grape Skin: A Feasible Ingredient for the Formulation of Snacks with Potential for Reducing the Risk of Diabetes.

In the present work the feasibility of Tannat grape skin (TGS) as a functional ingredient in the formulation of two snacks (yogurt and biscuits) was studied. The research provided novel information on the effects of the food matrix and digestion process, under simulated human oral gastrointestinal conditions, in the bioaccessibility of TGS bioactive compounds composing of the snacks with health promoting properties (antioxidant, anti-inflammatory, and antidiabetic). TGS polyphenolic profile was analyzed by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) finding mainly flavonoids, phenolic acids, and anthocyanins, which may exert antioxidant, anti-inflammatory, and carbohydrase inhibition capacities. TGS digest showed antioxidant and antidiabetic potential compared to the undigested sample (p < 0.05). Yogurt and biscuits with TGS were developed with the nutrition claims "no-added sugars" and "source of fiber" and were digested in vitro to evaluate the bioaccessibility of compounds with health promoting properties after food processing and digestion. After in vitro simulation of digestion, bioactive properties were enhanced for control and TGS snacks which may be attributed to the formation/release of compounds with health-promoting properties. Biscuits showed significant increase in ABTS antioxidant capacity and yogurt showed increased α-glucosidase inhibition capacity by the addition of TGS (p < 0.05). Polyphenols from TGS and bioactive peptides from snacks which may be released during digestion might be responsible for the observed bioactivities. Consumer's acceptance of TGS yogurt and biscuits showed scores of 6.3 and 5.1 (scale 1−9), respectively, showing TGS yogurt had higher overall acceptance. Sensory profile assessed by check-all-that-apply + just-about-right (CATA+JAR) showed most of the attributes were evaluated as "just about right", supporting good food quality. The developed yogurt presented adequate shelf-life parameters for 28 days. TGS yogurt with higher acceptability showed reduced ROS formation (p < 0.05) induced by tert-butyl hydroperoxide (1 mM) in CCD-18Co colon cells and RAW264.7 macrophages when pre-treated with concentrations 500−1000 and 100−500 µg/mL of the digests, respectively. Moreover, TGS yogurt digest pre-treatment reduced nitric oxide (NO) production (p < 0.05) in lipopolysaccharide (LPS)-induced RAW264.7 macrophages, showing anti-inflammatory potential. Bioactive peptides generated during lactic fermentation and digestion process may be contributors to intracellular effects. In conclusion, yogurt and biscuits with Tannat grape skin addition were obtained with nutrition claims "no-added sugars" and "source of fiber" with the potential to modulate key biochemical events associated with diabetes pathogenesis.

White rot fungal impact on the evolution of simple phenols during decay of silver fir wood by UHPLC-HQOMS.

INTRODUCTION: Silver fir (Abies alba Mill.) is one of the most valuable conifer wood species in Europe. Among the main opportunistic pathogens that cause root and butt rot on silver fir are Armillaria ostoyae and Heterobasidion abietinum. Due to the different enzymatic pools of these wood-decay fungi, different strategies in metabolizing the phenols were available. OBJECTIVE: This work explores the changes in phenolic compounds during silver fir wood degradation. METHODOLOGY: Phenols were analyzed before and after fungus inoculation in silver fir macerated wood after 2, 4 and 6 months. All samples were analyzed using high-performance liquid chromatography coupled to a hybrid quadrupole-orbitrap mass spectrometer. RESULTS: Thirteen compounds, including simple phenols, alkylphenyl alcohols, hydroxybenzoketones, hydroxycinnamaldehydes, hydroxybenzaldehydes, hydroxyphenylacetic acids, hydroxycinnamic acids, hydroxybenzoic acids and hydroxycoumarins, were detected. Pyrocatechol, coniferyl alcohol, acetovanillone, vanillin, benzoic acid, 4-hydroxybenzoic acid and vanillic acid contents decreased during the degradation process. Methyl vanillate, ferulic acid and p-coumaric were initially produced and then degraded. Scopoletin was accumulated. Pyrocatechol, acetovanillone and methyl vanillate were found for the first time in both degrading and non-degrading wood of silver fir. CONCLUSIONS: Despite differences in the enzymatic pool, both fungi caused a significant decrease in the amounts of phenolic compounds with the accumulation of the only scopoletin. Principal component analysis revealed an initial differentiation between the degradation activity of the two fungal species during degradation, but similar phenolic contents at the end of wood degradation.

Time course accumulation of polysulfides in Chardonnay and model juice fermentations.

Over the last five years, polysulfides in wine have become a topic of interest. The finding that both yeast and the wine composition could contribute to the formation and evolution of these compounds has prompted several studies, further corroborated by the discovery of a link between polysulfides and H2S release during ageing. In the present study, the accumulation of cysteinylated and glutathionylated polysulfides was followed during fermentation for the first time. Synthetic grape media and Chardonnay juice treated with CuSO4, elemental sulfur or both were fermented, and subsequently analysed using an in-house UHPLC-HRMS method. Differences in polysulfide accumulation were observed between the two media, highlighting the role of the matrix composition. Elemental sulfur, especially when combined with CuSO4, resulted in significantly increased accumulations of polysulfides compared to controls in both media. Polysulfides with longer S-chains generally appeared later in the fermentation, an observation that prompted further questions on these metabolites' formation.

Improvement of Caciotta-like cheese nutritional value by means of enrichment with blackcurrant (Ribes nigrum) and Cornelian cherry (Cornus mas).

Introduction: In this study, we supplemented models of Caciotta-like cheese with blackcurrant (Ribes nigrum) and Cornelian cherry (Cornus mas), as they have a high content of polyphenols, known as phytochemicals associated with health benefits. We evaluated the microbial composition, organoleptic aspects, total phenolic content, and chemical composition of model cheeses enriched with blackcurrant and Cornelian cherry. Methods: Two different suppliers have been tested: a conventional and an organic one. Two different conditions of preparation (freeze-dried and not freeze-dried) were tested in two different amounts (0.3 and 0.6% dry weight w/v milk volume). Polyphenols were determined using Folin-Ciocalteu reaction and spectrometry; microbial community was determined with selective 24 media and plate counts; composition was determined using nuclear magnetic resonance spectrometry. Organoleptic tests with an untrained panel have been performed. Results: The enrichments with blackcurrant and Cornelian cherry increased the total polyphenol content in model cheeses, in particular, when blackcurrant and Cornelian cherry were from conventional farming. Blackcurrant-enriched cheeses showed higher counts of lactic acid bacteria, higher levels of organic acids, amino acids, gamma-aminobutyric acid, histamine, and lower amount of monosaccharides deriving from bacterial lactose fermentation in cheese, suggesting a positive effect of blackcurrant compounds on the growth and activity of lactic acid bacteria. The enrichments did not affect the acceptance of the cheese, neither by blackcurrant nor by Cornelian cherry incorporation, with the exception of the appearance. Discussion: Overall, we showed that cheeses enriched with blackcurrant or Cornelian cherry from conventional farming increased the bioactive potential of the dairy product without having an adverse effect on the microbial community, physiochemical properties, or organoleptic properties.

In Vitro Bioaccessibility of Bioactive Compounds from Citrus Pomaces and Orange Pomace Biscuits.

The present investigation aimed to provide novel information on the chemical composition and in vitro bioaccessibility of bioactive compounds from raw citrus pomaces (mandarin varieties Clemenule and Ortanique and orange varieties Navel and Valencia). The effects of the baking process on their bioaccessibility was also assessed. Samples of pomaces and biscuits containing them as an ingredient were digested, mimicking the human enzymatic oral gastrointestinal digestion process, and the composition of the digests were analyzed. UHPLC-MS/MS results of the citrus pomaces flavonoid composition showed nobiletin, hesperidin/neohesperidin, tangeretin, heptamethoxyflavone, tetramethylscutellarein, and naringin/narirutin. The analysis of the digests indicated the bioaccessibility of compounds possessing antioxidant [6.6-11.0 mg GAE/g digest, 65.5-97.1 µmol Trolox Equivalents (TE)/g digest, and 135.5-214.8 µmol TE/g digest for total phenol content (TPC), ABTS, and ORAC-FL methods, respectively; significant reduction (p < 0.05) in Reactive Oxygen Species (ROS) formation under tert-butyl hydroperoxide (1 mM)-induced conditions in IEC-6 and CCD-18Co cells when pre-treated with concentrations 5-25 µg/mL of the digests], anti-inflammatory [significant reduction (p < 0.05) in nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 macrophages], and antidiabetic (IC50 3.97-11.42 mg/mL and 58.04-105.68 mg/mL for α-glucosidase and α-amylase inhibition capacities) properties in the citrus pomaces under study. In addition, orange pomace biscuits with the nutrition claims "no-added sugars" and "source of fiber", as well as those with good sensory quality (6.9-6.7, scale 1-9) and potential health promoting properties, were obtained. In conclusion, the results supported the feasibility of citrus pomace as a natural sustainable source of health-promoting compounds such as flavonoids. Unfractionated orange pomace may be employed as a functional food ingredient for reducing the risk of pathophysiological processes linked to oxidative stress, inflammation, and carbohydrate metabolism, such as diabetes, among others.

Stable Isotope Ratios of Herbs and Spices Commonly Used as Herbal Infusions in the Italian Market.

Stable isotope ratio analysis has been widely used for traceability and authenticity purposes in relation to various food commodities, but only in a limited number of herb and spice species. This study explored the stable isotope ratios of carbon, nitrogen, sulfur, oxygen, and hydrogen (δ13C, δ15N, δ34S, δ18O, and δ2H) of 119 herbs and spices belonging to 116 plant species and 57 plant families collected from the Italian market for the first time. The characteristic value ranges of δ13C, δ15N, δ34S, δ18O, and δ2H of the herbs and spices went from -31.0 to -11.6, -4.7 to 12.0, -5.0 to 22.0, 14.7 to 46.0, and -158 to -12‰, respectively. The isotopic profiles within and between common botanical families and their similarity/dissimilarity between herbs and spices belonging to the common botanical families are also discussed here. The results of this exploratory work highlight the possibility of characterizing herbs and spices and suggest widening the scope of the survey through more extensive sampling and focusing on specific plant species.

In Vitro Bioaccessibility of Extractable Compounds from Tannat Grape Skin Possessing Health Promoting Properties with Potential to Reduce the Risk of Diabetes.

Diabetes pathogenesis encompasses oxidative stress, inflammation, insulin malfunctioning and partial or total insulin secretion impairment, which leads to a constant hyperglycemia. Polyphenols are known to possess bioactive properties, being Tannat grape skin a natural and sustainable source of these compounds. The present study aimed to find out the bioaccessibility of health-promoting molecules composing a multifunctional extract from Tannat grape skin obtained under hydro-alcoholic-acid conditions. The identification of phenolic compounds in the samples was performed by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Subsequently, the samples were in vitro digested mimicking the human oral gastrointestinal conditions and the bioactivity of the digest (antioxidant, anti-inflammatory and modulation of glucose metabolism) was assessed. Effect on glucose metabolism was estimated by measuring carbohydrases activity and the functionality of glucose transporters of small intestine cells in presence and absence of the digested extract. Flavonoids, phenolic acids and phenolic alcohols were the major phenol compounds detected in the extract. The bioaccessible compounds protected the intestinal cells and macrophages against the induced formation of reactive oxygen species (ROS) and nitric oxide (NO). In addition, glucose transporters were inhibited by the digested extract. In conclusion, the bioaccessible compounds of the extract, including phenols, modulated key biochemical events involved in the pathogenesis of diabetes such as oxidative stress, inflammation and glucose absorption. The extract was effective under prevention with co-administration conditions supporting its potential for either reducing the risk or treating this disease.

A novel LC-HRMS method reveals cysteinyl and glutathionyl polysulfides in wine.

A novel ultra high pressure liquid chromatography combined with high resolution mass spectrometry (UHPLC-HRMS) method was developed to study glutathionyl and cysteinyl polysulfides in wine. Different HPLC columns were investigated in order to optimise the chromatographic resolution of the polysulfide standard mixtures synthesised in-house. The optimisation of the chromatographic conditions when trying to separate glutathionylated and cysteinylated species containing from 3 to 5 sulfur atoms proved particularly challenging, with the cationic exchange column IonPac CS12A-MS resulting to be the best column for this task.The synergistic application of the newly developed methods together with the synthesised reference standard mixtures allowed the identification and the detection of 11 different glutathionyl and cysteinyl polysulfides. Moreover, analysing 15 young white wines was possible to confirm the presence of GSSSG in wine (GS = glutathione). More importantly, this study allowed for the first identification of several symmetric and asymmetric new polysulfides, namely: GSSSSG, CSSSC (CS = cysteine), CSSSSC, CSSSG, and CSSSSG. These molecules have not previously been identified in wine, raising the question on their biogenesis and role on wine quality.

Identification and characterisation of thiolated polysulfides in must and wine using online SPE UHPLC-HRMS.

3-Mercaptohexanol and 4-methyl-4-mercaptopentanone are volatile thiols with a low perception threshold and are found in relatively low concentrations in several types of wines, e.g. Sauvignon blanc. They contribute positively to the flavour of a wine when in their free form, but they can oxidise, especially in the presence of Cu2+ and sulfur residues originating from pesticide treatments on the grapes. This condensation reaction results in the formation of polysulfides, which during storage can cause the release of H2S: a compound known to give rise to off-flavour in wine. The formation of these polysulfides has been proposed to originate from cysteinyl and glutathionyl S-conjugate precursors, but they have not yet been characterised. In this work, a method using ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry and online solid-phase extraction was designed and optimised for detection of these S-conjugates. The method allowed the detection of 21 suspected symmetrical and asymmetrical S-conjugates in thiol-supplemented aqueous solutions, of which 17 were also recovered in supplemented synthetic musts and wines. Moreover, the proposed method was used to investigate polysulfide formation upon addition of two different types of sulfur. Differences in formation of S-conjugates were evident in the synthetic samples: a higher relative abundance was observed upon addition of wettable sulfur compared to washed sulfur. For the commercial wines, differences in polysulfide formation were minor and merely related to the differences between the wines and not to the type of sulfur added.

Aphid-infested beans divert ant attendance from the rosy apple aphid in apple-bean intercropping.

Ecological intensification of cropping systems aims at restoring multi-functionality while supporting current productivity levels. Intercropping is a form of ecological intensification involving ecological processes beneficial to farmers that do not take place in monocultures. Thus, it represents a practical approach to decrease the use of synthetic inputs such as insecticides in cultivated systems. Whereas insecticide reduction via intercropping-facilitated suppression of aphids is reported in literature, the majority of published studies focussed on herbaceous crops. Thus, the effect of intercropping on aphid populations of cultivated trees remains largely unaddressed. In this study we hypothesized that intercropping a specific companion plant within perennial crops would divert ant attendance from an aphid attacking the crop to another aphid feeding on the newly introduced plant, reducing aphid damage on the crop. We tested our hypothesis in the system of apple (Malus domestica Borkhausen), the rosy apple aphid (Dysaphis plantaginea Passerini) and the black garden ant (Lasius niger L.). Bean plants (Vicia faba) with the black bean aphid (Aphis fabae Scopoli) were intercropped within apple trees inoculated with D. plantaginea. We measured ant attendance, aphid development and survival as well as honeydew composition on both plant species through semi-field and field experiments. The majority of ants chose to attend A. fabae over D. plantaginea in the semi-field experiment with potted plants. In the orchard, a larger majority of scouts were scored on A. fabae over D. plantaginea. A higher number of D. plantaginea colonies remained active in the apple control, whilst they were almost eradicated by intercropping. Although chemical analyses of honeydew disclosed differences in the carbohydrate and amino acid profiles between aphid species, the difference in honeydew composition did not explain the preference for A. fabae. Ants did not discriminate between the two honeydew mimics both in laboratory and field bioassays. Our results showed the potential of intercropping apple trees with beans as a method to reduce ant attendance and thus colony survival. We propose that intercropping represents a bottom-up approach towards ecological intensification of perennial crops. Together with other ecosystem-based measures such as habitat management, intercropping should be considered when planning ecosystem redesign to increase biological control of pests.

Production of Naturally γ-Aminobutyric Acid-Enriched Cheese Using the Dairy Strains Streptococcus thermophilus 84C and Lactobacillus brevis DSM 32386.

The cheese-derived strains Streptococcus thermophilus 84C isolated from Nostrano cheese, and Lactobacillus brevis DSM 32386 isolated from Traditional Mountain Malga cheese have been previously reported as γ-aminobutyric acid (GABA)-producers in vitro. In the present study, the ability of these strains to produce GABA was studied in experimental raw milk cheeses, with the aim to investigate the effect of the culture and the ripening time on the GABA concentration. The cultures used consisted on S. thermophilus 84C alone (84C) or in combination with L. brevis DSM 32386 (84C-DSM). The control culture was a commercial S. thermophilus strain, which was tested alone (CTRL) or in combination with the L. brevis DSM 32386 (CTRL-DSM). The pH evolution, microbiological counts, MiSeq Illumina and UHPLC-HQOMS analysis on milk and cheese samples were performed after 2, 9, and 20 days ripening. During the whole ripening, the pH was always under 5.5 in all batches. The concentration of GABA increased during ripening, with the highest content in 84C after 9 days ripening (84 ± 37 mg/kg), in 84C-DSM and CTRL-DSM after 20 days ripening (91 ± 28 and 88 ± 24 mg/kg, respectively). The data obtained support the hypothesis that S. thermophilus 84C and L. brevis DSM 32386 could be exploited as functional cultures, improving the in situ bio-synthesis of GABA during cheese ripening.