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Mattia Pia ARENA

Ricercatore t.d. art. 24 c. 3 lett. A
Dipartimento di Scienze della Vita sede ex-Agraria

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2024 - Oxidative fermentation of glucose and ethanol in designed media and cooked grape must by acetic acid bacteria [Articolo su rivista]
Brugnoli, M.; Cantadori, E.; Arena, M. P.; Gullo, M.

In this study, acetic acid bacteria strains were investigated for their ability to oxidize different carbon sources producing the corresponding oxidative products. Bacterial strains were cultivated in seven designed media and their acetification ability was assessed. The most performing strains were further tested to evaluate gluconic acid production in cooked grape must. Organic acids, sugars, and ethanol concentrations were assayed by high-pressure liquid chromatography. Overall, the findings showed high variability amongst strains of the same species, especially amongst Gluconobacter oxydans strains. However, strains ATCC 621H and DSM 3503T resulted to be the highest gluconic acid producers in all tested conditions. This study shows that grape must can be further valorized by selective fermentations for the production of gluconic based products.

2023 - Zero- and Low-Alcohol Fermented Beverages: A Perspective for Non-Conventional Healthy and Sustainable Production from Red Fruits [Articolo su rivista]
Brugnoli, Marcello; Cantadori, Elsa; Arena, Mattia Pia; DE VERO, Luciana; Colonello, Andrea; Gullo, Maria

The growing health consciousness among consumers is leading to an increased presence of functional foods and beverages on the market. Red fruits are rich in bioactive compounds such as anthocyanins with high antioxidant activity. In addition, red fruits contain sugars and are rich in phenolic compounds, vitamin C, dietary fibers, and manganese. Due to these characteristics, they are also suitable substrates for fermentation. Indeed, nowadays, microbial transformation of red fruits is based on alcoholic or lactic fermentation, producing alcoholic and non-alcoholic products, respectively. Although products fermented by acetic acid bacteria (AAB) have been thoroughly studied as a model of health benefits for human beings, little evidence is available on the acetic and gluconic fermentation of red fruits for obtaining functional products. Accordingly, this review aims to explore the potential of different red fruits, namely blackberry, raspberry, and blackcurrant, as raw materials for fermentation processes aimed at producing low- and no-alcohol beverages containing bioactive compounds and no added sugars. AAB are treated with a focus on their ability to produce acetic acid, gluconic acid, and bacterial cellulose, which are compounds of interest for developing fruit-based fermented beverages.

2020 - From Microbial Ecology to Innovative Applications in Food Quality Improvements: the Case of Sourdough as a Model Matrix. [Articolo su rivista]
Arena, M; Russo, P; Spano, G; Capozzi, V

Since millennia, humankind has exploited microbial diversity associated to give foodmatrices in order to obtain fermented foods and beverages, resulting in products with improvedquality and extended shelf life. This topic has received deserved and continuous interest in thescientific community, for the reason of its significance as a driver of innovation in the food and beveragesector. In this review paper, using sourdough as a model matrix, we provide some insights into thefield, testifying the relevance as a transdisciplinary subject. Firstly, we encompassed the prokaryoticand eukaryotic microbial diversity associated with the sourdough ecosystems. The importance ofthis micro-biodiversity in the light of flour-related chemical diversity was examined. Finally, wehighlighted the increasing interest in microbial-based applications oriented toward biocontrol solutionin the field of sourdough-based products (i.e., bread).

2018 - Exopolysaccharides Produced by Lactic Acid Bacteria and Their Role in the Food Industry [Capitolo/Saggio]
Arena, M. P.; Russo, P.; Spano, G.; Capozzi, V.; Fiocco., D.

Bacteria can produce a wide range of biopolymers (mainly polyesters, polyamides, inorganic polyanhydrides, and polysaccharides), with significant and heterogeneous physiological functions in prokaryotes (Rehm, 2010). Several of these compounds find application in the industrial and medical field, often offering—because of their biocompatibility and biodegradability—sustainable solutions from the economic, environmental, and social point of view (Rehm, 2010; Ates, 2015).

2018 - Small heat shock proteins characterization in a probiotic model [Abstract in Atti di Convegno]
Arena, Mp; Longo, A; Capozzi, V.; Spano, G.; Fiocco, D.

Small heat shock proteins (sHSP) are ubiquitous, low molecular weight proteins with chaperon-like activity. sHSP protect cell under stress conditions, critically contributing to survival to heat shock. Lactobacillus plantarum (Lp), a member of the lactic acid bacteria (LAB), is a probiotic species with biomedical and biotechnological applications. Unlike most lactobacilli, which have single shsp genes, three sHSP-encoding genes were identified in Lp WCFS1. Such redundancy might endow Lp with the capacity to cope with a broad range of stresses, thus accounting for its extraordinary environmental adaptability. To unravel the role of Lp sHSP, knock out (KO) mutants for hsp1 and hsp3 were generated and phenotypically characterized. Growth and survival rates under diverse stress conditions, which are typical for probiotics, revealed a different contribute of the two sHSP to thermotolerance induction and cryprotection. However, neither hsp seemed essential to tackle such challenges. Accordingly, cellular protein aggregation in both mutants was not significantly different from the wild type. Comparative transcriptional patterns revealed that in the mutant genetic backgrounds there is an up-regulated basal expression of the un-mutated mate hsp and other stress-related genes genes, which may compensate for the loss of sHSP function, hence underlying the lack of a marked susceptibility to stress. hsp KO affected biofilm adhesive capacity, changed cell surface physicochemical properties, and drastically modified membrane fluidity upon stress. These findings indicate that hsp1 and hsp3 have pleiotropic effects, fulfill overlapping activities in stress tolerance and housekeeping functions, and regulate membrane fluidity by a plausible direct association.

2017 - β-glucans and probiotics [Articolo su rivista]
Arena, MATTIA PIA; Spano, Giuseppe; Fiocco, Daniela

In the recent decades, several health-promoting features have been ascribed to β-glucans. Biological properties such as anti-cancer, antiinflammatory and immune-modulating activity have been claimed for these polysaccharides. Moreover, when β-glucans were used in association with probiotic bacteria, prebiotic effects have been demonstrated, due to their ability to enhance growth, metabolism and/or beneficial activities of probiotics. This review shall present an overview of those studies, which have documented the diverse health-benefits of β-glucans and their association with probiotics.

2017 - Lactobacillus plantarum with broad antifungal activity: A promising approach to increase safety and shelf-life of cereal-based products [Articolo su rivista]
Russo, Pasquale; Arena, MATTIA PIA; Fiocco, Daniela; Capozzi, Vittorio; Drider, Djamel; Spano, Giuseppe

Cereal-based fermented products are worldwide diffused staple food resources and cereal-based beverages represent a promising innovative field in the food market. Contamination and development of spoilage filamentous fungi can result in loss of cereal-based food products and it is a critical safety concern due to their potential ability to produce mycotoxins. Lactic Acid Bacteria (LAB) have been proposed as green strategy for the control of the moulds in the food industry due to their ability to produce antifungal metabolites. In this work, eighty-eight Lactobacillus plantarum strains were screened for their antifungal activity against Aspergillus niger, Aspergillus flavus, Fusarium culmorum, Penicillium roqueforti, Penicillium expansum, Penicillium chrysogenum, and Cladosporium spp. The overlayed method was used for a preliminary discrimination of the strains as no, mild and strong inhibitors. L. plantarum isolates that displayed broad antifungal spectrum activity were further screened based on the antifungal properties of their cell-free supernatant (CFS). CFSs from L. plantarum UFG 108 and L. plantarum UFG 121, in reason of their antifungal potential, were characterized and analyzed by HPLC. Results indicated that lactic acid was produced at high concentration during the growth phase, suggesting that this metabolic aptitude, associated with the low pH, contributed to explain the highlighted antifungal phenotype. Production of phenyllactic acid was also observed. Finally, a new oat-based beverage was obtained by fermentation with the strongest antifungal strain L. plantarum UFG 121. This product was submitted or not to a thermal stabilization and artificially contaminated with F. culmorum. Samples containing L. plantarum UFG 121 showed the best biopreservative effects, since that no differences were observed in terms of some qualitative features between not or contaminated samples with F. culmorum. Here we demonstrate, for the first time, the suitability of LAB strains for the fermentation and antifungal biopreservation of oat-based products.

2017 - The potential of lactic acid bacteria to colonize biotic and abiotic surfaces and the investigation of their interactions and mechanisms [Articolo su rivista]
Arena, MATTIA PIA; Capozzi, Vittorio; Spano, Giuseppe; Fiocco, Daniela

Lactic acid bacteria (LAB) are a heterogeneous group of Gram-positive bacteria that comprise several species which have evolved in close association with humans (food and lifestyle). While their use to ferment food dates back to very ancient times, in the last decades, LAB have attracted much attention for their documented beneficial properties and for potential biomedical applications. Some LAB are commensal that colonize, stably or transiently, host mucosal surfaces, inlcuding the gut, where they may contribute to host health. In this review, we present and discuss the main factors enabling LAB adaptation to such lifestyle, including the gene reprogramming accompanying gut colonization, the specific bacterial components involved in adhesion and interaction with host, and how the gut niche has shaped the genome of intestine-adapted species. Moreover, the capacity of LAB to colonize abiotic surfaces by forming structured communities, i.e., biofilms, is briefly discussed, taking into account the main bacterial and environmental factors involved, particularly in relation to food-related environments. The vast spread of LAB surface-associated communities and the ability to control their occurrence hold great potentials for human health and food safety biotechnologies.

2016 - Chemical composition and the anti-melanogenic potential of different essential oils [Articolo su rivista]
Fiocco, Daniela; Arciuli, Marcella; Arena, Mattia Pia; Benvenuti, Stefania; Gallone, Anna

Tyrosinase is the key enzyme of melanogenesis, the process that determines skin and hair pigmentation in mammals. In humans, various pigmentary disorders depend on excessive melanin production and accumulation. Moreover, tyrosinase activity can cause the undesirable browning of plant-derived food. For such reasons, ongoing researches aim at developing effective melanogenesis inhibitors for medical, cosmetic and food applications. Diverse biological activities have been observed for certain plant essential oils (EOs) and their components. In this study, the EOs from nine different plant species were investigated for their anti-melanogenic potential. The EOs chemical composition was assessed by gas chromatography-mass spectrometry (GC-MS) revealing the presence of some constituents with known tyrosinase inhibitory ability. Most EOs inhibited significantly and dose-dependently mushroom tyrosinase. Two samples also inhibited the activity of tyrosinase from murine B16 melanoma cells. Our findings indicate that some of the analysed EOs may be promising natural sources for developing novel anti-melanogenic agents, thus having a good potential in medical, cosmetic and food-processing industries.

2016 - Combinations of cereal β-glucans and probiotics can enhance the anti-inflammatory activity on host cells by a synergistic effect [Articolo su rivista]
Arena, MATTIA PIA; Russo, Pasquale; Capozzi, Vittorio; Rascón, Ana; Felis, Giovanna E.; Spano, Giuseppe; Fiocco, Daniela

The influence of oat and barley β-glucans on some features of diverse probiotic strains was investigated. Neither barley nor oat β-glucans supported the growth of any of the investigated strains when used as the sole carbon source in fermentation media. No protective effect by β-glucans was observed on bacterial survival to in vitro simulation of the human digestive tract. Moreover, the presence of β-glucans did not enhance the ability of probiotic bacteria to adhere on human intestinal cells. The immunomodulatory activities of probiotics, β-glucans, and their combinations were studied and compared in vitro by transcriptional analysis of immune-related genes on LPS-stimulated THP-1 macrophages. Immune modulating properties were evidenced. In particular, mixtures of probiotic microorganisms and barley β-glucans exhibited synergistic effects in modulating the transcriptional level of several immune-related genes, leading to an overall enhanced anti-inflammatory effect. Together, these findings suggest a promising application of probiotic bacteria and β-glucans in the preparation of dietary mixtures presenting health-promoting features such as immunomodulatory properties.

2016 - β-Glucans and Synbiotic Foods [Capitolo/Saggio]
Arena, MATTIA PIA; Russo, Pasquale; Fiocco, Daniela; Capozzi, Vittorio; Spano, Giuseppe

The beneficial effects of probiotic, prebiotic, and synbiotic on human health are increasingly expanding their claim. Concurrently with the selection and characterization of microorganisms is selected and characterized their for probiotic features, at the same time there is a growing interest to individuate and study novel prebiotic compounds aiming to develop novel synbiotic formulations which hold health-promoting properties. In the view of what has been earlier reported, this work includes a comprehensive overview of β-glucans compounds that are currently the focus of numerous research studies due to their attractive biological activity and prebiotic characteristics.

2016 - Industrial microorganisms: tolerance to antibiotics and application of antimicrobial agents [Capitolo/Saggio]
Arena, MATTIA PIA; Russo, Pasquale; Fiocco, Daniela; Spano, Giuseppe

Lactic acid bacteria (LAB) are a group of microorganisms generally associated to food industry as they are traditionally applied in the production of several fermented food. Beside the aim to optimize the quality of the end product determining the organoleptic and nutritional profile, LAB can also contribute to the safety assessment of food producing different compounds with antimicrobial activity such as bacteriocins, antimicrobial peptides and low molecular weight compounds. The production of antimicrobial compounds could be considered a suitable trait to be sought in the selection of starter, non-starter and probiotic strains due to the realistic use of these bacteria in the food industry in order to reduce spoilage and pathogenic microorganisms, increase the shelf life, moderate or eliminate the chemical additives. Moreover, the antagonistic activity can also aim to contrast the alarming problem of antibiotic resistance that is a crucial safety pre-assessment. Here, the potential use of LAB as biopreservatives and biotherapeutic agents in the food industry, and an overview about their resistance to antibiotics are discussed.

2016 - Starter Cultures for Sparkling Wine [Articolo su rivista]
Garofalo, C; Arena, M; Laddomada, B; Cappello, Ms; Grieco, F; Beneduce, L; Spano, G; Capozzi, V

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.

2016 - Stressors and Food Environment: Toward Strategies to Improve Robustness and Stress Tolerance in Probiotics [Capitolo/Saggio]
Capozzi, Vittorio; Arena, MATTIA PIA; Russo, Pasquale; Spano, Giuseppe; Fiocco, Daniela

The stress encountered by probiotics in food during manufacturing, processing, and storage can reduce their survival, stability, and functionality, thus having a negative impact on the performance of related functional food. A clear knowledge of the stress factors threatening probiotic viability in food, understanding the microbial cell mechanisms underlying stress tolerance, and investigations on how stress resistance may be ameliorated, can contribute to the development of strategies to enhance the potential health benefits of probiotic food. The recent evidence suggesting a consistent influence of the food matrix on the probiotic properties sheds new light on the field of food-related probiotic stress response. In addition, the improvement of probiotic robustness might lead to reconsideration of the use of probiotic strains with interesting health properties but with poor technological characteristics.

2016 - The lactic acid bacteria of fermented fruits and vegetables [Capitolo/Saggio]
Russo, Pasquale; Caggianiello, Graziano; Arena, MATTIA PIA; Fiocco, Daniela; Capozzi, Vittorio; Spano, Giuseppe

Vegetables fermented foods and beverages are traditional worldwide consumed foodstuffs, since fermentation is one of the most important methods of food preservation still in use for vegetables and fruits. Several species of lactic acid bacteria are involved in the fermentation of vegetable matrices. These microorganisms transform raw materials trough their metabolism conferring specific nutritional and organoleptic properties to the product. The industrial demand of standardization of the product and the process, as well as the need to ensure food safety, require to drive a controlled fermentation. Therefore, selected cultures of lactic acid bacteria are recommended as microbial starter for industrial fermentations. In the last years, an increasing interest has been generated about the beneficial role played by lactic acid bacteria in the gut environment. For this reason, fermented food and beverages from vegetables have been proposed as optimal carriers to deliver probiotic bacteria and for the elaboration of functional foods. In this chapter, we shall provide a background on the key role of lactic acid bacteria in vegetable fermentations, focusing on both traditional and innovative applications for the food industry.

2016 - The Microbiota of Non-cow Milk and Products [Capitolo/Saggio]
Alexandraki, V.; Kazou, M.; Angelopoulou, A.; Arena, MATTIA PIA; Capozzi, Vittorio; Russo, Pasquale; Fiocco, Daniela; Spano, Giuseppe; Papadimitriou, K.; Tsakalidou, E.

Milk, a basic component of the human diet, is rich in all kinds of nutrients, such as proteins, carbohydrates, lipids, vitamins, and minerals. These compounds comprise an ideal growth substrate for both harmful and beneficial microorganisms, making milk a perishable raw material. As a result, milk contains numerous microorganisms that originate either from the animal itself, even when the animal is clinically healthy, or from the environment during the collection and storage of milk. The increasing social, economic, and research on non-cow types of milk and its products illustrates the renewed interest in the microbiota associated with these matrices. The present work is an overview of existing knowledge on the microbiota of sheep, goat, buffalo, camel, equine, yak, and human milk and their products. Relevant studies concerning both classical microbiological approaches as well as advanced molecular methods are included.

2016 - Use of Lactobacillus plantarum strains as a bio-control strategy against food-borne pathogenic microorganisms [Articolo su rivista]
Arena, MATTIA PIA; Silvain, Amandine; Normanno, GIOVANNI GIUSEPPE; Grieco, Francesco; Drider, Djamel; Spano, Giuseppe; Fiocco, Daniela

Lactobacillus plantarum is one of the most versatile species extensively used in the food industry both as microbial starters and probiotic microorganisms. Several L. plantarum strains have been shown to produce different antimicrobial compounds such as organic acids, hydrogen peroxide, diacetyl, and also bacteriocins and antimicrobial peptides, both denoted by a variable spectrum of action. In recent decades, the selection of microbial molecules and/or bacterial strains able to produce antagonistic molecules to be used as antimicrobials and preservatives has been attracting scientific interest, in order to eliminate or reduce chemical additives, because of the growing attention of consumers for healthy and natural food products. The aim of this work was to investigate the antimicrobial activity of several food-isolated L. plantarum strains, analyzed against the pathogenic bacteria Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus. Antagonistic activity was assayed by agar spot test and revealed that strain L. plantarum 105 had the strongest ability to contrast the growth of L. monocytogenes, while strains L. plantarum 106 and 107 were the most active microorganisms against E. coli O157:H7. The antimicrobial ability was also screened by well diffusion assay and broth micro-dilution method using cell-free supernatants (CFS) from each Lactobacillus strain. Moreover, the chemical nature of the molecules released in the CFS, and possibly underlying the antagonistic activity, was preliminary characterized by exposure to different constraints such as pH neutralization, heating, catalase, and proteinase treatments. Our data suggest that the ability of L. plantarum cultures to contrast pathogens growth in vitro depends, at least in part, on a pH-lowering effect of supernatants and/or on the presence of organic acids. Cluster analysis was performed in order to group L. plantarum strains according to their antimicrobial effect. This study emphasizes the tempting use of the tested L. plantarum strains and/or their CFS as antimicrobial agents against food-borne pathogens.

2015 - Functional Starters for Functional Yogurt. [Articolo su rivista]
Arena, Mattia P.; Caggianiello, Graziano; Russo, Pasquale; Albenzio, Marzia; Massa, Salvatore; Fiocco, Daniela; Capozzi, Vittorio; Spano, Giuseppe

In this study, we investigated the multifunctionality (microbial starters and probiotics) of Lactobacillus plantarum WCFS1 and Lactobacillus plantarum CECT 8328 strains used as microbial starters for the production of yogurt in combination with Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus. The ability of the probiotic strains to survive to oro-gastrointestinal stresses was monitored by an in vitro assay simulating the human digestive tract. The transcriptional level of several genes involved in the immune response suggested that the probiotic strains may have a favorable influence on immunomodulation. Overall, this study revealed that the tested lactobacilli exhibited suitable technological features for yogurt production and might be used to formulate novel food with immunomodulating effects.

2015 - Nuovo ceppo di Lactobacillus [Brevetto]
Russo, Pasquale; Arena, MATTIA PIA; Capozzi, Vittorio; Fiocco, Daniela; Spano, Giuseppe

2014 - Ananas di IV gamma come alimento funzionale per veicolare batteri lattici probiotici [Relazione in Atti di Convegno]
Russo, P.; de Chiara, M. L. V.; Vernile, A.; Amodio, M. L.; Arena, M. P.; Capozzi, V.; Spano, G

2014 - Antimicrobial activity of lactic acid bacteria on biofilm formed by Listeria monocytogenes [Abstract in Atti di Convegno]
Spano, Giuseppe; Arena, MATTIA PIA; Capozzi, Vittorio; A., Hadjilouka; S., Paramithiotis; E. F., Drosinos; Russo, Pasquale

Lactic acid bacteria are proposed as an innovative eco-friendly strategy to control relevant food-borne pathogens including Listeria monocytogenes and L. monocytogenes represent a safety concern for the food industry due to its ability to survive and grow under several harsh conditions associated with food processing and preservation. In addition, L. monocytogenes may persist in food plants and equipment since able to form biofilms. In the present work, we screened 152 lactic acid bacteria for their ability to inhibit the growth of L. monocytogenes serovar ½a and 4b from animal and vegetable origin. The antagonistic effect was investigated by analysing the halo of inhibition on agar plates co-inoculated with LAB strains or in presence of the correspondent cell-free supernatant. The same approach was employed to evaluate the reduction of biofilm formation on glass, polystirene, and stainless steel. L. monocytogenes biofilms were quantified spectrophotometrically and the viability of the pathogen assessed by qPCR. LAB were clustered in four categories according to their inhibitor effect. The strains with the stronger antagonistic activity suggest a potential employment to control L. monocytogenes proliferation and the corresponding biofilm formation in food processing and plants.

2014 - Antimicrobial activity of Lactobacillus plantarum strains and antimicrobial properties of L. plantarum small heat shock proteins [Abstract in Atti di Convegno]
Spano, G.; Arena, M. P.; Caggianiello, G.; Capozzi, V.; Russo, P.; Fiocco, D.

The aim of this study was to investigate Lactobacillus plantarum strains possessing antimicrobial features to be proposed as potential bioagents and biopreservant stains that could prevent the growth of pathogenic bacteria in fermented food. A total of 85 Lactobacillus plantarum strains, including strains overproducing small heat shock proteins, were investigated for their antimicrobial activity against the pathogenic bacteria Escherichia coli O157:H7, Staphylococcus aureus, Listeria monocytogenes and Salmonella enterica. Firstly, the antimicrobial effect was evaluated using over-night cultures (ONC) of lactobacilli by agar spot test. The antagonistic effects of L. plantarum strains against pathogens were detected measuring the growth inhibition halos on plates. All strains showed significant antibacterial activity and we classified L. plantarum strains as no, mild, strong and very strong inhibiting strain when inhibitions zone of 1 mm, 2 mm, 2-5 mm and more than 5 mm were produced. Subsequentially, 15 strains showing strong and very strong antimicrobial activity were selected and their cell-free supernatant (CFS) was harvested and used for agar diffusion and microdilution bioassays over an incubation time of 24 h at 30 °C, in order to investigate the substances determining the antimicrobial ability. Overall, the results showed an interesting antagonism depending both on L. plantarum strain analysed and pathogens bacteria tested. Moreover, antimicrobial activity was observed by L. plantarum strains overproducing small heat shock proteins. The applications of L. plantarum strains exhibiting antimicrobial activity need to be considered as safety bio-alternative to traditional antibiotics used against human infection and to chemical substances used in the food processing.

2014 - Barley β-Glucans-Containing Food Enhances Probiotic Performances of Beneficial Bacteria [Articolo su rivista]
Arena, Mattia P.; Caggianiello, Graziano; Daniela, Fiocco; Pasquale, Russo; Torelli, Michele; Giuseppe, Spano; Vittorio, Capozzi

Currently, the majority of prebiotics in the market are derived from non-digestible oligosaccharides. Very few studies have focused on non-digestible long chain complex polysaccharides in relation to their potential as novel prebiotics. Cereals β-glucans have been investigated for immune-modulating properties and beneficial effects on obesity, cardiovascular diseases, diabetes, and cholesterol levels. Moreover, β-glucans have been reported to be highly fermentable by the intestinal microbiota in the caecum and colon, and can enhance both growth rate and lactic acid production of microbes isolated from the human intestine. In this work, we report the effects of food matrices containing barley β-glucans on growth and probiotic features of four Lactobacillus strains. Such matrices were able to improve the growth rate of the tested bacteria both in unstressed conditions and, importantly, after exposure to in vitro simulation of the digestive tract. Moreover, the effect of β-glucans-containing food on bacterial adhesion to enterocyte-like cells was analyzed and a positive influence on probiotic-enterocyte interaction was observed.

2014 - Comparative genome analysis reveals strains specific gene clusters involved in exopolysaccharides biosynthesis in Lactobacillus plantarum [Abstract in Atti di Convegno]
Spano, G.; Caggianiello, G.; Fiocco, D.; Arena, M. P.; Russo, P.; Orrù, L.; Lamontanara, A.; van Hijum, S.; Capozzi, V.

Several lactic acid bacteria (LAB) produce capsular and secreted forms of exopolysaccharides (EPS) (Remus et al., 2012). The EPS production is correlated to the presence of specific gene clusters (eps/cps) which exhibit a conserved modular organization and include genes encoding both regulatory factors and enzymes involved in EPS biosynthesis, polymerization and secretion, including glycosyl-transferases, which are responsible for the assembly of the characteristic EPS-repeating unit. The Lactobacillus plantarum WCFS1 genome encodes 4 cps clusters of genes that are associated with surface polysaccharide production. The cps2A-J and cps4A-J clusters encode all functions required for capsular polysaccharide formation, while the cps1A-I and cps3A-J clusters lack genes encoding chain-length control functions and a priming glycosyl-transferase (Remus et al., 2012). L. plantarum Lp90 exhibited a ropy phenotype which was associated to its capacity to produce EPS. The genetic basis of EPS production was investigated by identification, sequencing and comparative analysis of genes clusters involved in EPS biosynthesis with other L. plantarum genomes. It was found that the cluster 4 (cps4) is the most conserved, while the cluster 3 presents high homology with cps3 of WCFS1, ST-III and ATCC 14917 L. plantarum strains. Conversely, cluster 1 (cps1) is fully present only in L. plantarum JDM1, while the first genes of cluster 2 are homologous to the cps2A-E genes of other L. plantarum. The remaining genes of cluster 2 are homologous to proteins of Lactobacillus fabifermentans T30PCM01, probably involved in the exopolysaccharides biosynthesis. This feature makes unique the organization of cps clusters L. plantarum Lp90 and may explain the ropy phenotype of this strain.

2014 - Fresh-cut pineapple as a new carrier of probiotic lactic acid bacteria [Articolo su rivista]
Russo, Pasquale; Maria Lucia Valeria de, Chiara; Vernile, Anna; Amodio, MARIA LUISA; Arena, MATTIA PIA; Spano, Giuseppe; Capozzi, Vittorio

Due to the increasing interest for healthy foods, the feasibility of using fresh-cut fruits to vehicle probiotic microorganisms is arising scientific interest. With this aim, the survival of probiotic lactic acid bacteria, belonging to Lactobacillus plantarum and Lactobacillus fermentum species, was monitored on artificially inoculated pineapple pieces throughout storage. The main nutritional, physicochemical and sensorial parameters of minimally processed pineapples were monitored. Finally, probiotic Lactobacillus were further investigated for their antagonistic effect against Listeria monocytogenes and Escherichia coli O157:H7 on pineapple pieces. Our results show that at eight-days of storage, the concentration of L. plantarum and L. fermentum on pineapples pieces ranged between 7.3 and 6.3 log cfu g-1 respectively, without affecting the final quality of the fresh-cut pineapple. The antagonistic assays indicated that L. fermentum was able to inhibit the growth of both pathogens, while L. plantarum was effective only against L. monocytogenes. This study suggest that both L. plantarum and L. fermentum could be successfully applied during processing of fresh-cut pineapples, contributing at the same time to inducing a protective effect against relevant foodborne pathogens.

2014 - Lactobacillus plantarum as a Strategy for an In situ Production of Vitamin B2 [Articolo su rivista]
Arena, MATTIA PIA; Fiocco, Daniela; Massa, Salvatore; Capozzi, Vittorio; Russo, Pasquale; Spano, Giuseppe

The European Food Safety Agency (EFSA) has recently introduced a system for a pre-market safety assessment of selected taxonomic groups of microorganisms leading to a Qualified Presumption of Safety (QPS), European equivalent of the Generally Recognized as Safe (GRAS) status. Several species of food-related lactic acid bacteria (LAB) associated with food, have obtained a QPS status. The adaptability of LAB to fermentation process, their biosynthetic capacity and metabolic versatility, are some of the principal features that facilitate the application of LAB in foods for producing, releasing and/or increasing specific beneficial compounds. Among these, vitamin production by LAB has recently gained attention of the scientific community. The proper selection and exploitation of nutraceutical-producing LAB is an interesting strategy to produce novel fermented foods with increased nutritional and/or health promoting properties. Fermented milks or bread with high levels of B-group vitamins (such as folate and riboflavin) can be produced by LAB-promoted biosynthesis. In this paper, the use of probiotics bacteria such as Lactobacillus plantarum as a strategy for an in situ production and/or overproduction of vitamin B2 will be discussed.

2014 - Probiotic abilities of riboflavin-overproducing Lactobacillus strains: a novel promising application of probiotics [Articolo su rivista]
Arena, Mattia P.; Russo, Pasquale; Capozzi, Vittorio; López, Paloma; Fiocco, Daniela; Spano, Giuseppe

The probiotic potential of Lactobacillus plantarum and Lactobacillus fermentum strains, capable of overproducing riboflavin, was investigated. The riboflavin production was quantified in co-cultures of lactobacilli and human intestinal epithelial cells, and the riboflavin overproduction ability was confirmed. When milk and yogurt were used as carrier matrices, L. plantarum and L. fermentum strains displayed a significant ability to survive through simulated gastrointestinal transit. Adhesion was studied on both biotic and abiotic surfaces. Both strains adhered strongly on Caco-2 cells, negatively influenced the adhesion of Escherichia coli O157:H7, and strongly inhibited the growth of three reference pathogenic microbial strains. Resistance to major antibiotics and potential hemolytic activity were assayed. Overall, this study reveals that these Lactobacillus stains are endowed with promising probiotic properties and thus are candidates for the development of novel functional food which would be both enriched in riboflavin and induce additional health benefits, including a potential in situ riboflavin production, once the microorganisms colonize the host intestine.

2014 - Probiotic attributes of riboflavin over-producing lactic acid bacteria strains [Abstract in Atti di Convegno]
Spano, G.; Arena, M. P.; Fiocco, D.; Russo, P.; Capozzi, V.

Riboflavin plays a crucial role in human cell metabolism as precursor of coenzymes flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) (Capozzi et al., 2012). In this study, the riboflavin production of Lactobacillus plantarum and Lactobacillus fermentum strains was investigated in chemically defined medium (CDM) (Russo et al., 2014) and in a co-cultures system of lactobacilli and human intestinal epithelial cells (Russo et al., 2012). Both strains showed the ability to over-produce riboflavin respect to the control strains. The strains displayed a significant ability to survive to an in vitro gastrointestinal transit, an aptitude to form biofilm on abiotic surface, a strong capability to adhere on intestinal cells and to contrast the adhesion of Escherichia coli O157:H7. Furthermore, both strains showed antimicrobial activity on agar plate against three human pathogenic bacteria, i.e. Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enterica. Resistance to major antibiotics and potential hemolytic activity confirmed the safety assessment. This study aims to demonstrate the combination of probiotic features and riboflavin over-production by the investigated Lactobacillus plantarum and Lactobacillus fermentum strains and their potential application as endogenousriboflavin producers that couldenrichedthe riboflavin content both in novel fermented food and in situ in the host intestine.

2014 - Probiotic survival in symbiotic yogurt-like cereal-based beverage [Abstract in Atti di Convegno]
Spano, G.; Caggianiello, G.; Fiocco, D.; Arena, M. P.; Russo, P.; Parisi, F.; Capozzi, V.

The scientific literature and market trends testify to the growing emphasis on the development of dairy-like vegetable based products, including food matrices based on fruit, vegetables and cereals, able to meet the needs of consumers allergic/intolerant, on cholesterol-restricted diets, and/or vegans. Together with the formulations based on soy flour, yogurt-like products produced from cereal flours, have been receiving increasing attention due to the good balance of texture, flavor, and nutritional properties in the final product. As substitutes of dairy productions, the corresponding non-dairy foods yogurt represent a novel alternative for the consumption of probiotic bacteria. In order to produce yogurt-like cereal-based beverages, a mixture of rice, barley, and oat flours and concentrated red grape must were used. In addition to standard formulation, we prepared a β-glucans enriched beverage using a flour of barley selected for its β-glucans content (12 g of β-glucans per 100 g of flour). β-glucans have been reported to be highly fermentable by the intestinal microbiota in the caecum and colon, and can enhance both growth rate and lactic acid production of microbes isolated from the human intestine. Throughout the technological phases, a commercial yogurt starter cultures and a selected strain of Lactobacillus plantarum were separately used to perform lactic acid fermentation (30 °C, 8 hours). During the storage at 4 °C, we assessed the survival of 7 probiotic strains ( Lactobacillus acidophilus LA5, L. plantarum CECT 8328, Lactobacillus johnsonii CECT 289, Lactobacillus fermentum CECT 8448, Lactobacillus reuteri CECT 925, Lactobacillus paracasei subsp. paracasei LC-01 , L. plantarum WCFS1) in the standard yogurt-like cereal-based beverage and in β-glucans enriched beverage (symbiotic yogurt-like cereal-based beverage).

2014 - Riboflavin-overproducing strains of Lactobacillus fermentum for riboflavin-enriched bread [Articolo su rivista]
Russo, Pasquale; Capozzi, Vittorio; Arena, MATTIA PIA; Spadaccino, Giuseppina; María Teresa, Dueñas; Paloma, López; Fiocco, Daniela; Spano, Giuseppe

Lactobacillus fermentum isolated from sourdough was able to produce riboflavin. Spontaneous roseoflavinresistant mutants were obtained by exposing the wild strain (named L. fermentum PBCC11) to increasing concentrations of roseoflavin. Fifteen spontaneous roseoflavin-resistant mutants were isolated, and the level of vitamin B2 was quantified by HPLC. Seven mutant strains produced concentrations of vitamin B2 higher than 1 mg L−1. Interestingly, three mutants were unable to overproduce riboflavin even though they were able to withstand the selective pressure of roseoflavin. Alignment of the rib leader region of PBCC11 and its derivatives showed only point mutations at two neighboring locations of the RFN element. In particular, the highest riboflavinproducing isolates possess an A to G mutation at position 240, while the lowest riboflavin producer carries a T to A substitution at position 236. No mutations were detected in the derivative strains that did not have an overproducing phenotype. The best riboflavin overproducing strain, named L. fermentum PBCC11.5, and its parental strain were used to fortify bread. The effect of two different periods of fermentation on the riboflavin level was compared. Bread produced using the coinoculum yeast and L. fermentum PBCC11.5 led to an approximately twofold increase of final vitamin B2 content

2013 - Essential oils as tyrosinase inhibitors: a different effect on tyrosinase of different sources. A preliminary study [Abstract in Atti di Convegno]
Fiocco, Daniela; M., Arciuli; Arena, MATTIA PIA; Capozzi, Vittorio; S., Benvenuti; A., Gallone

2013 - Inducciòn de crecimento de L. casei 393 despùs del proceso digestivo in vitro [Relazione in Atti di Convegno]
Melendez, Paola; Arena, MATTIA PIA; Spano, Giuseppe

gastric stress, Lactobacillus casei

2013 - Intraspecific riboflavin overproduction variability in lactic acid bacteria [Relazione in Atti di Convegno]
Russo, Pasquale; Spano, Giuseppe; Fiocco, Daniela; Capozzi, Vittorio; Arena, Mattia Pia

2013 - Nuovo ceppo di lactobacillus [Brevetto]
Russo, Pasquale; Capozzi, Vittorio; Fiocco, Daniela; Arena, MATTIA PIA; Spano, Giuseppe

2013 - Potential probiotic properties of riboflavin-overproducing Lactic Acid Bacteria [Poster]
Arena, MATTIA PIA; Russo, Pasquale; Capozzi, Vittorio; P., López; Spano, Giuseppe; Fiocco, Daniela

2011 - Effect of abiotic stress conditions on expression of the Lactobacillus brevis IOEB 9809 tyrosine decarboxylase and agmatine deiminase genes [Articolo su rivista]
Arena, Mp; Russo, Pasquale; Capozzi, Vittorio; Beneduce, Luciano; Spano, Giuseppe

Lactobacillus brevis IOEB 9809, isolated from red wine, is able to produce both tyramine and putrescine. Using a real-time quantitative reverse transription PCR (qRT-PCR) we analyzed the relative expression of L. brevis IOEB 9809 tdc and aguA1 genes, encoding, respectively, a tyrosine decarboxylase and an agmatine deiminase enzyme. Relative gene expression was monitored either during the different growth phases under optimal conditions or under abiotic stress commonly found in wine. The tdc and aguA1 specific cDNA were amplified with specific primers. Our results indicate that tdc and aguA1 genes are differently expressed during the different growth phases and transiently induced by ethanol (12% v/v) and acidic stresses (pH 3.2 and 5.0).

2011 - Expression of Lactobacillus brevis IOEB 9809 tyrosine decarboxylase and agmatine deiminase genes in wine is correlated to substrate availability [Articolo su rivista]
Arena, MATTIA PIA; Romano, A.; Capozzi, Vittorio; Beneduce, Luciano; Ghariani, M.; Grieco, F.; Lucas, P.; Spano, Giuseppe

Aims: Lactobacillus brevis IOEB 9809 is able to produce both tyramine and putrescine via tyrosine decarboxylase and agmatine deiminase enzymes, respectively, when cultured on synthetic media. The aims of this study were to assess the expression of L. brevis IOEB 9809 tdc and aguA1 genes, during wine fermentation and to evaluate the effect of substrate availability and pH on tdc and aguA1 expression, as well as on biogenic amine production and L. brevis viability. Methods and Results: The relative expression of L. brevis IOEB 9809 tdc and aguA1 genes was analysed in wine by quantitative real-time RT-PCR (qRTPCR) during a period of incubation of 30 days. Cell viability, pH values, putrescine and tyramine concentration were monitored throughout the experiments. Conclusions: The wine trials indicated that L. brevis IOEB 9809 is able to produce both tyramine and putrescine during wine fermentation. Increased cell viability was also observed in wine supplemented with tyrosine or agmatine. qRT-PCR analysis suggests a strong influence of substrate availability on the expression of genes coding for tyrosine decarboxylase and agmatine deiminase in L. brevis IOEB 9809. Less evident is the relationship between putrescine and tyramine production and tolerance to wine pH. Significance and Impact of Study: To our knowledge, this study represents the first assessment of relative expression of L. brevis IOEB 9809 genes involved in biogenic amine production in wine. Furthermore, an effect of biogenic amine production on viability of L. brevis during wine fermentation was established.

2011 - The hsp 16 gene of the probiotic Lactobacillus acidophilus is differently regulated by salt, high temperature and acidic stresses as revealed by reverse transcription quantitative PCR (qRT-PCR) analysis [Articolo su rivista]
Capozzi, Vittorio; Arena, Mattia Pia; Crisetti, Elisabetta; Spano, Giuseppe; Fiocco, Daniela

Small heat shock proteins (sHsps) are ubiquitous conserved chaperone-like proteins involved in cellular proteins protection under stressful conditions. In this study, a reverse transcription quantitative PCR (RT-qPCR) was developed and used to quantify the transcript level of a small heat shock gene (shs) in the probiotic bacterium Lactobacillus acidophilus NCFM, under stress conditions such as heat (45 °C and 53 °C), bile (0.3% w/v), hyperosmosis (1 M and 2.5 M NaCl), and low pH value (pH 4). The shs gene of L. acidophilus NCFM was induced by salt, high temperature and acidic stress, while repression was observed upon bile stress. Analysis of the 5' noncoding region of the hsp 16 gene reveals the presence of an inverted repeat (IR) sequence (TTAGCACTC-N9-GAGTGCTAA) homologue to the controlling IR of chaperone expression (CIRCE) elements found in the upstream regulatory region of Gram-positive heat shock operons, suggesting that the hsp16 gene of L. acidophilus might be transcriptionally controlled by HrcA. In addition, the alignment of several small heat shock proteins identified so far in lactic acid bacteria, reveals that the Hsp16 of L. acidophilus exhibit a strong evolutionary relationship with members of the Lactobacillus acidophilus group.

2010 - Are consumers aware of the risks related to Biogenic Amines in food? [Capitolo/Saggio]
Russo, P.; Spano, Giuseppe; Arena, M. P.; Capozzi, Vittorio; Fiocco, Daniela; Grieco, F.; Beneduce, Luciano

in press

2010 - Different regulation of the tyrosine decarboxylase and the agmatine deiminase genes in Lactobacillus brevis IOEB 9809 [Articolo su rivista]
Arena, M. P.; Russo, P.; Vittorio, Capozzi; Giuseppe, Spano