The variation in the coding capacity within Oenococcus oeni can have a significant impact on wine quality. The detection of several genes involved in important metabolic pathways (i.e. citrate, sulphur and arginine metabolisms) was performed on 10 indigenous O. oeni strains from Negroamaro wine, a red table wine (Apulia, Italy). These strains were selected from 95 isolates, collected during spontaneous malolactic fermentation, according to the results of an Amplified Fragment Length Polymorphism (AFLP) analysis. A total of 16 genes were screened, most (11) of which had never previously been assayed on O. oeni. All strains possessed 10 genes encoding enzymes such as malolactic enzyme (mleA), esterase (estA), citrate lyase (citD, citE and citF), citrate transporter (maeP), alpha-acetolactate decarboxylase (alsD), alpha-acetolactate synthase (alsS), S-adenosylmethionine synthase (metK) and cystathionine beta-lyase (metC) and resulted negative in the detection of genes encoding cystathionine gamma-lyase (metB), ornithine transcarbamylase (arcB) and carbamate kinase (arcC). The sequence of PCR fragments of 11 genes of a representative strain (ITEM 15929) was compared to those of three reference O. oeni strains. The indigenous strain was phylogenetically more similar to PSU-1 and ATCC BAA1163 than AWRI B429. This study describes new genetic markers useful for detecting the genetic potential of O. oeni strains to contribute to aroma production and for investigating the population structure of the species. (C) 2014 Elsevier Ltd. All rights reserved.

Alginate micro beads containing Lactobacillus kefiri (the principal bacteria present in the kefir probiotic drink)were produced by a novel technique based on dual aerosols spaying of alginate based solution and CaCl2 ascross linking agent. Carboxymethylcellulose (CMC) has been also added to the alginate in order to change thephysic-chemical properties (viscosity and permeability) of the microbeads. Calcium alginate and CMC are biopolymersthat can be used for developing oral drug-delivery systems. These biopolymers have been reportedto show a pH-dependent swelling behaviour. Calciumalginate and CMC have also been known to possess an excellentmucoadhesive property. The loaded microbeads have been characterized in terms of morphology, chemicalcomposition and stability in different conditions mimicking the gastric environment.In this study, we demonstrate the feasibility of a continuous fabrication of alginate microbeads in a range of50-70 ?m size, encapsulating L. kefiri as active ingredient. The technique involves the use of a double aerosols ofalginate based solution and CaCl2 as crosslinking agent. Moreover, the encapsulation process was proved to beeffective and not detrimental to bacteria viability. At the same time, it was verified the protective efficacy ofthe microcapsules against the gastric environment using both SGF pH 1.2 (fasted state) and pH 2.2 (feed state).

The bioactivities of Santolina corsica Jord. & Fourr. n-hexane (EHS) and methanol (EMS) extracts were evaluated in relation to their chemical profile. Main methods: EHS and EMS were analysed by gas chromatography-mass spectrometry and high performance liquid chromatography-diode array detection (HPLC-DAD), respectively. Antioxidant activity was determined by ?-carotene bleaching, Ferric Reducing Activity Power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2?-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) tests. Nitric oxide (NO) production was assessed in LPS-stimulated RAW 264.7 cells. Anti-proliferative activity was evaluated by MTT assay on A549, HeLa, PC3, MCF-7, MDA-MB-231 cancer cells, and non-tumorigenic MCF10 A cells. Cell motility, migration and invasion were assessed by wound-healing scratch, migration and invasion assays, respectively. DNA fragmentation was tested by TUNEL assay. Cells morphology was studied by phase-contrast microscopy. Procaspase-8, -9, poly (ADP-ribose) polymerase and COX-2 expression levels were evaluated by immunoblotting analysis. Key findings: Kaempferol-3-O-glucoside (5878.67 mg/100 g of extract), chlorogenic acid (746.11 mg/100 g), and rosmarinic acid (550.16 mg/100 g) were the dominant EMS constituents. EHS showed myrcene (18.86%) as the main compound, followed by palmitic acid methyl and ethyl esters (9.35 and 9.16%, respectively), ?-phellandrene (8.48%), and ar-curcumene (5.63%). Both extracts showed promising anti-proliferative activity on all tested cancer cells, without inducing cytotoxicity in non-tumorigenic cells MCF-10 A. Moreover, extracts inhibited motility, migration, and invasion of MDA-MB-231 cells, inducing apoptosis. EHS decreased NO production, showing anti-inflammatory activity. Significance: S. corsica extracts might be potentially useful in cancer treatment, since reduce invasive and migratory potential of MDA-MB-231 cells triggering apoptosis.

Glycyrrhiza glabra cultivation and harvesting produces substantial quantities of aerial parts as waste. With the aim to prospect an innovative valorization of these byproducts, the aerial parts were harvested in May and October and analyzed for their chemical profile, antioxidant properties, and effects on viability of five cancer cell lines. Pinocembrin was the main constituent. A significant protection of lipid permddation was observed with the May total extract (IC50 of 4.2 +/- 0.4 mu g/mL at 30 min of incubation). The effects on viability of HeLa, MCF-7, MDA-MB-231, Caco-2, and PC3 human cancer cells were investigated. All samples shown a remarkable activity with IC50 values below 25 mu g/mL. Samples from plants harvested in May exhibited greater activity than those harvested in October. MCF-7 and HeLa were the most sensitive cells with IC50 in the range 2.73-3.01 and 3.28-5.53 mu g/mL, respectively. G. glabra aerial parts represent a good source of valuable products.

Oenococcus oeni is the most important LAB species involved in MLF. Highgenotypic heterogeneity of O. oeni strains and a further divergence within thebacterial strain population has been demonstrated. Genotypic diversityseems to be correlated with different metabolic characteristics of bacteria andthis may affect the organoleptic properties of obtained wines. The aim ofthis work was to detect several genes involved in important metabolic pathways(i.e. citrate, sulphur and arginine metabolisms) in 10 indigenous O. oeni strainsselected from Negroamaro wine, a red table wine (Apulia, Italy).This work revealed several new genetic markers that made possible to use a PCRdetection approach to investigate strain heterogeneity for a large range ofgenes encoding enzymes of oenological relevance.

There is scarce information on the occurrence of several fungi that infect witheredgrapes to produce passito wine. Isolation and characterization of Neofusicoccum parvumstrains and evaluation of their effects on withered grape and wine were carried out.Methods and Results: Nine isolates were phenotypically characterized by colonymorphology and genetically discriminated by molecular methods. Two representative strainswere identified as N. parvum according to phylogenetic analysis of internal transcribed spacer (ITS), and a part of translation elongation factor 1-alfa (TEF) and ?-tubulin DNA sequences.The pathogenicity of both strains on grape berries varied according to the inoculation andincubation conditions. Under withering conditions, infected berries showed browning andshriveling and some berries showed pycnidial development on the surface. The infectionaffected laccase, esterase, ?-glucosidase and tannase on grape juice as well as the content ofseveral aroma molecules on resulting wines. Strain-specific effects on wine composition werealso observed.Conclusions: N. parvum occurred in withered grapes and was able to infect grapes underwithering condition changing the aroma wine.Significance and Impact of the Study: This study reports for the first time the N. parvumisolation in fruit-drying rooms and indicates its important role on post-harvest grapeinfection.

In the last two decades knowledge on lactic acid bacteria (LAB) associated with wine has increased considerably. Investigations on genetic and biochemistry of species involved in malolactic fermentation, such as Oenococcus oeni and of Lactobacillus have enabled a better understand of their role in aroma modification and microbial stability of wine. In particular, the use of molecular techniques has provided evidence on the high diversity at species and strain level, thus improving the knowledge on wine LAB taxonomy and ecology. These tools demonstrated to also be useful to detect strains with potential desirable or undesirable traits for winemaking purposes. At the same time, advances on the enzymatic properties of wine LAB responsible for the development of wine aroma molecules have been undertaken. Interestingly, it has highlighted the high intraspecific variability of enzymatic activities such as glucosidase, esterase, proteases and those related to citrate metabolism within the wine LAB species. This genetic and biochemistry diversity that characterizes wine LAB populations can generate a wide spectrum of wine sensory outcomes. This review examines some of these interesting aspects as a way to elucidate the link between LAB diversity with wine aroma and flavour. In particular, the correlation between inter- and intra-species diversity and bacterial metabolic traits that affect the organoleptic properties of wines is highlighted with emphasis on the importance of enzymatic potential of bacteria for the selection of starter cultures to control MLF and to enhance wine aroma.

A simple procedure is proposed for selective proteinsolubilization and trypsin digestion, followed by off-line liquid chromatography-matrix assisted laser desorption ionization mass spectrometry (LC-MALDI MS) analysis of Oenococcus oeni (O. oeni) bacterium. Peptides wereidentified from tryptic digests using sequencing by tandem massspectrometry and database searches. Cytoplasmic and membrane relatedproteins (MRP) were identified in the O. oeni bacterium. MS/MS dataanalysis points out 13 peptides having one point mutation from 9 proteins.The major microheterogeneity was found for Zn-dependent alcoholdehydrogenase (Zn-ADH, Q04GE6) and 60 kDa chaperonin (GroEL,Q04E64) that are involved in methionine catabolism and post-translationalprotein folding, respectively. MS/MS data processing also leads to theidentification of 34 unique phosphorylation sites from 19 phosphoproteins.

The diversity of indigenous Oenococcus oeni strains was investigated by molecular and biochemical characterization of isolates from Malvasia Nera wine, an economically important red wine of the Salento Region (Apulia, Italy), during spontaneous malolactic fermentation (MLF). A total of 82 isolates of this species, identified by species-specific PCR and 16S rDNA sequence analysis, were molecularly characterized by the Amplified Fragment Length Polymorphism (AFLP) technique. Three main groups resulted by clustering analysis and showed intraspecific homology higher than 50% and a total of seven subgroups, with similarity values ranged from 80 to 98%, were obtained within these groups. Enzymatic activities, such as esterase, ²-glucosidase, protease, and the consumption rate of L-malic acid, citric acid, acetaldehyde and arginine were assessed in the representative strains according to AFLP analysis. The results displayed different enzymatic activities and consumption rates of tested metabolites among the strains. No correlation between molecular and biochemical data was observed. The evidence of biochemical variability observed among ‘Malvasia Nera’ strains demonstrated that the wine aroma can be modulated depending on the strains involved in the MLF. Hence, the heterogeneity existing within natural O. oeni populations represents an interesting ecological source that can be useful for technological purposes.

Actually, the knowledge about wine yeasts remains largely dominated by the extensive studies on Saccharomyces (S.) cerevisiae. Molecular methods, allowing discrimination of both species and strains in winemaking, can profitably be applied for characterization of the microflora occurring in winemaking and for monitoring the fermentation process. Recently, some novel yeast isolates have been described as hybrid between S. cerevisiae xSaccharomyces species, leaving the Saccharomyces strains containing non-Saccharomyces hybrids essentially unexplored. In this study, we have analyzed a yeast strain isolated from “Primitivo” grape (http://www.ispa.cnr.it/index.php?page=collezioni&lang=en accession number 12998) and we found that, in addition to the S. cerevisiae genome, it has acquired genetic material from a non-Saccharomyces species. The study was focused on the analysis of chromosomal and mitochondrial gene sequences (ITS and 26S rRNA, SSU and COXII, ACTIN-1 and TEF), 2D-PAGE mitochondrial proteins, and spore viability. The results allowed us to formulate the hypothesis that in the MSH199 isolate a DNA containing a rDNA sequence from Hanseniaspora vineae, a non-Saccharomyces yeast, was incorporated through homologous recombination in the grape environment where yeast species are propagated. Moreover, physiological characterization showed that the MSH199 isolate possesses high technological quality traits (fermentation performance) and glycerol production, resistance to ethanol, SO2 and temperature) useful for industrial application.

The purpose of this study was to perform the molecular and technological characterization of indigenous strains of Oenococcus oeni, with excellent winemaking skills, isolated from wines produced by grapes of two different Salento areas (Apulia, Italy), in order to select strains from using as a malolactic fermentation (MLF) starter in wine. Sequences discriminating the different species were obtained by species-specific PCR with primers specific for the MLE gene and subsequent sequence analysis of PCR products on RNA containing the region for the 16S gene. In this way were identified as O. oeni, 75 lactic acid bacteria from a total of 210, isolated from Negro amaro wine (Salice Salentino area) and 80 from a total of 220 lactic acid bacteria, isolated from Negro amaro wine (Guagnano area), during malolactic fermentation. Genotypic analysis was performed by molecular technique (AFLP) based on amplified fragment length polymorphism, in order to develop molecular profiles that differentiate and characterize several O. oeni strains.To design specific bacterial strains for the different characteristics of a wine, it is necessary to consider various criteria as resistance to ethanol, SO2, growth at low pH, assessment of the malate metabolism and absence of negative traits to the health of consumers (biogenic amines [BA], ethyl carbamate). These selection criteria were applied to several strains (chosen from different subgroups of the cluster obtained by AFLP). The results allowed to select some of these strains that may have a good fermentation performance.

Oenococcus oeni is the main lactic acid bacterium involved in Malolactic Fermentation since its high adaptation capacity in wine (1). Extensive studies, carried out over the years, furnished a considerable amount of information on the genetic, physiology and metabolism of this bacterium (2-4). The increasing acquisition of data about the secondary metabolic activities exhibited by O. oeni, which impact strongly to sensory properties of wine, stimulated the investigations on variability within indigenous populations isolated from various winemaking environments.Indeed, strains can modified differently the flavour, quality and safety of wine according to their to metabolic diversity and, due to the economic importance of wine, great interest has been addressed to the study of intraspecific heterogeneity of O. oeni (5, 6).In this work, the detection of several genes involved on important metabolic pathways (i.e. citrate, sulphur and arginine metabolism) was performed on 10 indigenous O. oeni strains of Negroamaro wine, a table red wine (Apulia, Italy). These strains were selected from 95 isolates, collected during a spontaneous malolactic fermentation, according to the results of Amplified Fragment Length Polymorphism (AFLP) analysis. It was screened a total of 16 genes, most of them (11) never assayed before on O. oeni. All strains possessed 10 genes encoding enzymes such as malolactic enzyme (mleA), esterase (estA), citrate lyase ?citD, citE and citF), citrate transporter (maeP), ?-acetolactate decarboxylase (alsD), ?-?acetolactatesynthase (alsS), S-adenosylmethionine synthase (metK) and cystathionine ?-lyase (metC) and resulted negative in the detection of genes encoding cystathionine ?-lyase (metB), ornithine transcarbamylase (arcB) and carbamate kinase (arcC) (table 1.). The sequence of PCR fragments of 11 genes of a representative strain (ITEM 15929) were compared to those of three reference O. oeni strain. The indigenous strain phylogenetically resulted more similar to PSU-1 and ATCC BAA1163 than AWRI B429. The present study provides information on population structure of the species and describes new genetic markers useful for detecting the genetic potential of O. oeni strains to contribute to aroma production and to improve the quality of wine.Table 1. Results of PCR detection of different enzyme-encoding genes in a population of 10 O.oeni strains.Target geneAccession number protein activities

Table olives are one of the most important traditional fermented vegetables in Europe and their world consumption is constantly increasing. In the Greek style, table olives are obtained by spontaneous fermentations, without any chemical debittering treatment. Evolution of sugars, organic acids, alcohols, mono and polyphenol compounds and volatile compounds associated with the fermentative metabolism of yeasts and bacteria throughout the natural fermentation process of the two Italian olive cultivars Cellina di Nardò and Leccino were determined. A new protocol was developed and applied aimed at the technological characterization of LAB and yeast strains as possible candidate autochthonous starters for table olive fermentation from Cellina di Nardò and Leccino cultivars. The study of the main physical, chemical and aromatic parameters during fermentation helped to determine chemical descriptors that may be suitable for monitoring olive fermentation. In both the analyzed table olive cultivars, aldehydes proved to be closely related to the first stage of fermentation (30 days), while higher alcohols (2-methyl-1-propanol; 3-methyl-1-butanol), styrene, and o-cymene were associated with the middle stage of fermentation (90 days) and acetate esters and acetic acid with the final step of olive fermentation (180 days).

Filamentous fungi are the main pathogens of withered grapes destined for passito wine production. Knowledge of which species inhabit these post-harvest fruits and their pathogenicity is essential in order to develop strategies to control infection, but is still scarce. This study investigated the predominant mycobiota of withered grapes through a cultivation-dependent approach. Strain and species heterogeneity was evidenced on examining isolates collected over three consecutive years. Colony morphology and PCR-restriction fragment length polymorphism (PCR-RFLP) analysis revealed the occurrence of several phenotypes and haplotypes, respectively. Strains were phylogenetically analyzed based on sequence typing of different genes or regions (e.g. calmodulin, ?-tubulin and internal transcribed spacer region). Beside the most common necrotrophic-saprophytic species of Penicillium, Aspergillus, Alternaria and Botrytis species responsible for fruit rot, other saprobic species were identified (e.g. Trichoderma atroviride, Sarocladium terricola, Arthrinium arundinis and Diaporthe eres) generally not associated with post-harvest fruit diseases. Species such as Penicillium ubiquetum, Cladosporium pseudocladosporioides, Lichtheimia ramosa, Sarocladium terricola, Diaporthe nobilis, Bipolaris secalis, Paraconiothyrium fuckelii and Galactomyces reessii that had never previously been isolated from grapevine or grape were also identified. Moreover, it was not possible to assign a species to some isolates, while some members of Didymosphaeriaceae and Didymellaceae remained unclassified even at genus level. This study provides insights into the diversity of the epiphytic fungi inhabiting withered grapes and evidences the importance of their identification to understand the causes of fruit diseases. Finally, phylogenetic species delimitation furnished data of interest to fungal taxonomy.

Polyunsaturated Fatty Acids (PUFAs) are a class of natural compoundswith interesting biochemical effects on human health. Indeed, in addition totheir beneficial effects in brain and cardiovascular disorders, ?-3 LC-PUFAsupplementation can exert antineoplastic activity by triggering cell death inhuman cancer cells, either alone or in combination with conventional therapies.The aim of this review is, by analyzing the recent scientific literature, tohighlight the molecular mechanisms of Omega-3Polyunsaturated Fatty Acids(?-3 PUFAs) in antineoplastic events, and the effects of supplementing dietswith them during chemotherapy regimens. This analysis may provide specificinformation to carry out future pre-clinical and clinical studies aimed at a betteruse of omega-3polyunsaturated fatty acids in cancer therapy.

The anticancer properties of Santolina corsica Jord. & Fourr. n-hexane (EHS) and methanol (EMS) extracts were herein evaluated, in relation to their chemical profile. EHS and EMS were analysed by gas chromatography-mass spectrometry and high performance liquid chromatography-diode array detection, respectively. Quercetin-like molecules were the dominant EMS constituents, while EHS showed high concentrations of sesquiterpenes and fatty acid esters. The anti-proliferative activity of those extracts was evaluated on a wide panel of cancer cell lines. Interestingly, anti-proliferative activities of both extracts were found to be very specific for cancer cells, since their IC50 values measured on a non-tumorigenic breast epithelial cell line (MCF 10A) were remarkably higher than those found for all the tested cancer cell lines. Moreover, our results proved that they were able to activate the extrinsic apoptosis pathway, reducing motility, as well as invasive and migratory potential of MDA-MB-231 cells, so counteracting their metastatic behaviour. Concluding, EHS and EMS might be taken into account for potential therapeutic applications in cancer treatment.

The sparkling wine market has expanded in recent years, boosted by the increasing demand of the global market. As for other fermented beverages, technological yeasts and bacteria selected to design commercial starter cultures represent key levers to maximize product quality and safety. The increasing economic interest in the sector of sparkling wine has also implied a renewed interest in microbial resource management. In this review, after a brief introduction, we report an overview of the main characterization criteria in order to select Saccharomyces cerevisiae strains suitable for use as starter cultures for the production of base wines and to drive re-fermentation of base wines to obtain sparkling wines. Particular attention has been reserved to the technological characterization aspects of re-fermenting phenotypes. We also analysed the possible uses of selected non-Saccharomyces and malolactic strains in order to differentiate specific productions. Finally, we highlighted the main safety aspects related to microbes of enological interest and underlined some microbial-based biotechnological applications helpful to pursue product and process innovations. Overall, the sparkling wine industry may find a relevant benefit from the exploitation of the wide resources associated with vineyard/wine microbial diversity

Table olives are one of the most important traditional fermented vegetables in Southern European countries and their consumption is constantly increasing throughout the world. Today, the industrial production of black table olives is carried out by spontaneous fermentation processes which are not predictable and are strongly influenced by the autochthonous microflora, the physical-chemical conditions, the availability of fermentable substrates and salt content. Evolution of sugars, organic acids, alcohols, mono and polyphenol compounds and volatile compounds associated with the fermentative metabolism of yeasts and bacteria throughout the natural fermentation process of the two Italian olive cultivars Cellina di Nardò and Leccino were determined. A new protocol was developed and applied aimed at the selection of LAB and yeast strains as candidate autochthonous starters for table olive fermentation from Cellina di Nardò and Leccino cultivars.

The lysozyme of hen's egg white is used in winemaking to control spontaneous lactic acid bacteria (LAB). A total of eight LAB strains, isolated from grape must and wine, were used to assess the inhibitory effects of wine phenolics on lysozyme activity. The presence of phenolics, extracted from grape pomace, in growth medium reduced the mortality rate due to the lysozyme activity. This effect was especially clear in the case of strains belonging to Lactobacillus uvarum, Pediococcus parvulus and Oenococccus oeni, which are more sensitive to lysozyme than L. plantarum and L. hilgardii strains. Cell lysis assays carried out on four strains sensitive to lysozyme and Micrococcus lysodeikticus ATCC 4698, used as a reference strain, confirmed the inhibition of grape pomace phenolics on the muramidase. There was no interference from non-flavonoids, flavanols and flavonol compounds, when they were tested individually, on the lysozyme activity against the strains. Anthocyanins extracted from grape skins slightly inhibited the activity only against M. lysodeikticus. However, proanthocyanidins extracted from seed berries, strongly inhibited the lysozyme. In this extract, dimers were the predominant oligomers of flavan-3-ol. The study demonstrated that the effectiveness of lysozyme against LAB in red winemaking is related to the amount of low molecular weight proanthocyanidins that are released when the grapes are macerating.

Table olives are one of the most important traditional fermented vegetables in Southern European (Italy, Greece and Spain) countries. In the Greek-style production system, the fruits are placed directly into the brine, thus allowing the natural fermentation to take place. The spontaneous fermentations, that can last 8-12 months, are driven by mixed populations of microorganisms, mainly the epiphytic microbial population of yeasts and lactic acid bacteria (LAB) (Romero et al., 2004). At present, the industrial table olive process is not predictable and depends on the empirical experience of the producers. In order to avoid the unpredictability of the olive spontaneous fermentation, to improve the productive process and to constantly produce high-quality final products, the use of strains of LAB as starter cultures for olive production has been proposed (Sabatini and Marsilioet al., 2008; Panagou et al., 2008; Blana et al., 2014). However, in the last years, the importance and the potential applications of yeasts as starters for table olive processing has been recognized (Arroyo-López et al., 2008, 2012; Bevilacqua et al., 2012). Objectives: In the present work, we have studied the main physical, chemical and aromatic parameters of natural fermentations of Cellina di Nardò, Leccino, Kalamata and Conservolea table olives in order to determine chemical descriptors correlated to microbiological activities and the dynamics of microorganisms in order to select LAB and yeast strains as candidate autochthonous starter cultures. Conclusions: The identified chemical descriptors can be suitable to follow the trend and to control the outcome of the fermentation and a new protocol aimed to the selection of LAB and yeast strains as candidates autochthonous starters has been developed and applied (Bleve et al. 2014 a, b). Selected microbial starters have been successfully used to ferment olives in pilot and industrial-scale and a new method for table olive production has been set up (Bleve et al. 2103). The use of selected autochthonous starter cultures produced fermented table olives with improved organoleptic, sensorial and nutritional characteristics.

La Lattoria è una azienda zootecnica del Salento per la produzione di latte bovino, che ha investito in impianti automatizzati per il veicolamento alla mungitura delle bovine e la raccolta robotizzata del latte. In questo modo, il latte è di alta qualità, paucimicrobico, e, grazie ad una alimentazione varia e controllata, ben equilibrato in sostanza grassa.L'azienda produce uno yogurt cremoso, non sgrassato, disponibile al pubblico tramite un distributore automatico refrigerato, da cui i clienti si riforniscono nelle 24 h, oltre che allo spaccio aziendale e in alcuni punti vendita a Lecce. Il benessere degli animali e un buon livello di pulizia forniscono una materia prima di elevato gusto, per la produzione di yogurt. La collaborazione con ISPA-CNR è in corso per evidenziare punti di forza e criticità e ampliare i segmenti di mercato (ceppi starter, tipi di frutta, alimenti funzionali).