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Andrea PULVIRENTI

Professore Associato
Dipartimento di Scienze della Vita sede ex-Agraria


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Pubblicazioni

2023 - Non-thermal techniques and the “hurdle” approach: How is food technology evolving? [Articolo su rivista]
Bigi, Francesco; Maurizzi, Enrico; Quartieri, Andrea; De Leo, Riccardo; Gullo, Maria; Pulvirenti, Andrea
abstract

Food technology has played a crucial role since the beginning of human civilization. Throughout the centuries, the evolution of food processing has led to an increase of food safety and quality, enhancing the overall quality of human life. Lately, academic research and industries have gained awareness about the impact of conventional preservation technologies like heat sterilization and chemical preservatives on environment and economy, besides the detrimental effects on the organoleptic and nutritional quality of foods. This consciousness oriented the efforts towards more sustainable techniques, paving the way to a new “green era” of food technology.Scope and approach This work explores seven non-thermal technologies, describing their theoretical principles, mechanism of action, effect on microorganisms, advantages, and limitations. Besides, the concept of hurdle technology to overcome the criticisms related to single processing techniques is highlighted. Key findings and conclusions Non-thermal technologies have the potential to substitute conventional techniques for microbial inactivation, improving the safety and quality of food. The efficiency of each technique strongly relies on the process parameters (treatment intensity; exposure time), equipment (geometry; conformation), product (physical state; composition; viscosity; geometry), and microorganism characteristics (strain; concentration; growth phase; resistance mechanisms). In this sense, the hurdle approach allows to overcome the limitations related to the single technologies, broadening their efficiency and application range, and minimizing their impact on food quality. Further studies are recommended to better understand the mechanisms of mutual interaction among these techniques when combined together in specific conditions, in view of their scaling-up for commercial applications.


2023 - Toxic metals in Loggerhead sea turtles (Caretta caretta) stranded freshly dead along Sicilian coasts [Articolo su rivista]
Cammilleri, Gaetano; Galluzzo, Francesco Giuseppe; Pulvirenti, Andrea; Pantano, Licia; Calabrese, Vittorio; Gentile, Antonino; Cumbo, Valentina; Macaluso, Andrea; Macaluso, Vito; Vella, Antonio; Ferrantelli, Vincenzo
abstract

Abstract Background: The Loggerhead sea turtle (Caretta caretta) is a marine reptile belonging to a monophyletic group of chelonians. As these animals are long-lived, they have the ability to accumulate pollutants. Aim: To collect epidemiological data on toxic metals in marine Loggerhead sea turtles. Materials and Methods: Forty Loggerhead sea turtles comprising 25 males and 15 females stranded freshly dead between 2013 and 2018 along the coasts of Sicily, Southern Italy, were examined for arsenic, cadmium, and lead accumulation in muscle and adipose tissues by means of a validated ICP-MS method. A modified K index as a growth condition factor, namely Fulton's K index, was used. Samples were tested in duplicate. A Wilcoxon rank sum test was carried out to evaluate metal contents differences between muscle and adipose tissues and between genders. Results: The Fulton's K index suggested a good body condition of the C. caretta recovered with mean values of 5.34±3.40 (n=40; ±SD). Detectable concentrations of lead were found in 70% of the samples analysed with mean values of 0.65±1.67 mg/kg wet weight and 0.51±1.29 mg/kg wet weight in muscle and adipose tissues, respectively. No significant differences in arsenic, cadmium, and lead were detected between genders. In addition, no significant correlation was found between modified K index and concentrations of arsenic, cadmium, and lead. Clinical relevance: Findings on muscle and adipose tissues suggest chronic exposure of Caretta caretta to high concentrations of especially lead which might negatively affect health and welfare of these marine turtles although body condition was good.


2022 - Anti-Spoilage Activity and Exopolysaccharides Production by Selected Lactic Acid Bacteria [Articolo su rivista]
Iosca, Giovanna; De Vero, Luciana; Di Rocco, Giulia; Perrone, Giancarlo; Gullo, Maria; Pulvirenti, Andrea
abstract

In this study, eight lactic acid bacteria (LAB) strains, previously isolated from traditional and gluten-free sourdoughs, and selected for their potential in improving the sensory and rheological quality of bakery products, were screened against some common spoilage agents. The anti-mould activity was tested using strains of the species Fusarium graminearum, Aspergillus flavus, Penicillium paneum and Aspergillus niger. Regarding the antibacterial activity, it was assessed against four strains of the species Escherichia coli, Campylobacter jejuni, Salmonella typhimurium and Listeria monocytogenes. Furthermore, LAB strains were evaluated for their ability to produce exopolysaccharides, which are gaining considerable attention for their functional properties and applicability in different food industrial applications. A strain-specific behaviour against the moulds was observed. In particular, F. graminearum ITEM 5356 was completely inhibited by all the LAB strains. Regarding the antibacterial activity, the strains Leuconostoc citreum UMCC 3011, Lactiplantibacillus plantarum UMCC 2996, and Pediococcus pentosaceus UMCC 3010 showed wide activity against the tested pathogens. Moreover, all the LAB strains were able to produce exopolysaccharides, which were preliminarily characterized. The assessed features of the LAB strains allow us to consider them as promising candidates for single or multiple starter cultures for food fermentation processes.


2022 - Attivitá biostimolante di un idrolizzato proteico fermentato ottenuto da sottoprodotti della pesca Ligure (PROGETTO FISH). I Convegno di Orticoltura e Floricoltura. [Abstract in Atti di Convegno]
Caser, M; Ertani, A; Benedetti, P; Gervasio, S; Masino, F; Cocchiara, S; Pulvirenti, A; Bertelli, C; Scariot, V; Bertora, C.
abstract


2022 - Candidate Acetic Acid Bacteria Strains for Levan Production [Articolo su rivista]
Anguluri, Kavitha; LA CHINA, Salvatore; Brugnoli, Marcello; DE VERO, Luciana; Pulvirenti, Andrea; Cassanelli, Stefano; Gullo, Maria
abstract

In this study, twelve strains of acetic acid bacteria (AAB) belonging to five different genera were tested for their ability to produce levan, at 70 and 250 g/L of sucrose concentration, respectively. The fructan produced by the bacterial strains was characterized as levan by NMR spectroscopy. Most of the strains produced levan, highlighting intra- and inter-species variability. High yield was observed for Neoasaia chiangmaiensis NBRC 101099 T, Kozakia baliensis DSM 14400 T and Gluconobacter cerinus DSM 9533 T at 70 g/L of sucrose. A 12-fold increase was observed for N. chiangmaiensis NBRC 101099 T at 250 g/L of sucrose concentration. Levan production was found to be affected by glucose accumulation and pH reduction, especially in Ko. baliensis DSM 14400 T. All the Gluconobacter strains showed a negative correlation with the increase in sucrose concentration. Among strains of Komagataeibacter genus, no clear effect of sucrose on levan yield was found. Results obtained in this study highlighted the differences in levan yield among AAB strains and showed interdependence between culture conditions, carbon source utilization, and time of incubation. On the contrary, the levan yield was not always related to the sucrose concentration.


2022 - Clonal selection of wine yeasts with differential adsorption activities towards phenolics and ochratoxin A [Articolo su rivista]
Caridi, Andrea; Sidari, Rossana; Pulvirenti, Andrea; Blaiotta, Giuseppe; Ritieni, Alberto
abstract

To design a rapid, simple, and low-cost procedure for yeast selection with differential adsorption activities toward phenolics and ochratoxin A, 284 yeast strains were screened. This was done by evaluating the type of growth during grape must fermentation, acetic acid production on Chalk agar, H2S production on BiGGY agar, and spore-formation on acetate agar. After that initial step, 84 strains were pre-selected and further studied by Petri plate tests and to determine their wine-making ability in trials and evaluating their differential adsorption activities toward phenolics and ochratoxin A. Three yeast strains were selected based on the above evaluations. After confirming that they belonged to Saccharomyces cerevisiae species and were diploids, a spore clonal selection was performed. The strain Sc1741A_1D was selected and used in winemaking at six Calabrian wineries and found to be suitable as wine starter to improve quality and safety of red wines.


2022 - Effect of foliar fertlizers from fish by-product on growth and development of Lettuce (L. sativa)in green-house conditions [Poster]
Ben Hassine, M.; Caradonia, F.; Milc, J. A.; Caccialupi, G.; Masino, F.; Pulvirenti, A.; Francia, E.; Antonelli, A.
abstract

In Liguria Region, as in all coastal areas, fish represents a relevant part of human diet due to its high-quality protein content, low fat content, and importance as a source of vitamins and minerals (Prato and Biandolino, 2015). Therefore, in the contest of circular economy, the huge quantity of fish by-product produced could be a resource instead of an environmental issue, thus producing materials with high added value. Such an example are fertilizers or biostimulants obtained from fish by-product that can be used to increase the sustainability of crops cultivation. Under the frame of project “FISH – Fertilizzante Innovativo Suolo e Habitat” (PSR Regione Liguria 2014 – 2020 Misura 16), the effect of foliar fertilizers obtained from fish by-product were evaluated on the growth and development of lettuce (Lactuca sativa L. ‘Summerbel’) in green-house conditions.


2022 - Fatty Acids Composition of Stomach Oil of Scopoli’s Shearwater (Calonectris diomedea) from Linosa’s Colony [Articolo su rivista]
Galluzzo, Francesco Giuseppe; Cumbo, Valentina; Cammilleri, Gaetano; Calabrese, Vittorio; Pulvirenti, Andrea; Cicero, Nicola; Pantano, Licia; Mascetti, Antonietta; Lo Cascio, Giovanni; Bacchi, Emanuela; Macaluso, Andrea; Vella, Antonio; Seminara, Salvatore; Ferrantelli, Vincenzo
abstract

Calonectris diomedea is a Procellariforms seabird having a very representative colony in Linosa Island (Southern Italy). The adult forms of C. diomedea produce a pasty oil from their proven- triculus to feed their chicks during the rearing period. In this work, we examined the fatty acids composition of the stomach oil of C. diomedea from Linosa Island by gas chromatography with flame ionization detection (GC-FID). The samples were collected at 20 and 70 days after hatching. Twenty different fatty acids (FAs) were identified. Saturated fatty acids (SFA) were the most abundant in percentage (41.6%) at day 20 followed by polyunsaturated fatty acids (PUFA, 38.7%) and monounsat- urated fatty acids (MUFA, 19.7%). MUFAs were the most abundant in samples collected at day 70 (53.8%), followed by SFAs (36.6%) and PUFAs (9.8%). Oleic acid (C18:1ω9) in the samples on day 70 was 4 times higher than that in the samples on day 20. The Principal Component Analysis (PCA) verified a clear separation of the stomach oil samples in two groups, according to the day of sampling. The results obtained confirm the role of FAs analysis of stomach oil to understand the ecology and breeding behaviour of C. diomedea, highlighting a resemblance with signatures recorded in marine organisms of Linosa Island.


2022 - Secondary shelf life assessment of UHT milk and its potential for food waste reduction [Articolo su rivista]
Nicosia, Carola; Fava, Patrizia; Pulvirenti, Andrea; Licciardello, Fabio
abstract

A scientifically based assessment of the secondary shelf life (SSL) might help reducing food waste generation at the household level. The present study addressed the SSL of two UHT milk brands upon simulated domestic use and storage after the first opening. Microbiological, sensory and physico-chemical parameters were monitored during 16 days of domestic storage to determine the period after opening during which UHT milk is still suitable for consumption. According to specific microbial and sensory acceptability limits, results showed that UHT milk is suitable for consumption until 6–7 days after first opening, even when managed in the worst conditions. The SSL of UHT milk may be extended from the current 3–4 days to 6–7 days, according to a precautionary approach. The reassessment of the labeled SSL of staple foods, such as milk, will contribute to reduce domestic food wastes, to increase sustainability of the food industry and competitiveness of food companies.


2022 - The Green Era of Food Packaging: General Considerations and New Trends [Articolo su rivista]
Maurizzi, Enrico; Bigi, Francesco; Quartieri, Andrea; De Leo, Riccardo; Volpelli, Luisa Antonella; Pulvirenti, Andrea
abstract

Recently, academic research and industries have gained awareness about the economic, environmental, and social impacts of conventional plastic packaging and its disposal. This consciousness has oriented efforts towards more sustainable materials such as biopolymers, paving the way for the “green era” of food packaging. This review provides a schematic overview about polymers and blends of them, which are emerging as promising alternatives to conventional plastics. Focus was dedicated to biopolymers from renewable sources and their applications to produce sustainable, active packaging with antimicrobial and antioxidant properties. In particular, the incorporation of plant extracts, food-waste derivatives, and nano-sized materials to produce bio-based active packaging with enhanced technical performances was investigated. According to recent studies, bio-based active packaging enriched with natural-based compounds has the potential to replace petroleum-derived materials. Based on molecular composition, the natural compounds can diversely interact with the native structure of the packaging materials, modulating their barriers, optical and mechanical performances, and conferring them antioxidant and antimicrobial properties. Overall, the recent academic findings could lead to a breakthrough in the field of food packaging, opening the gates to a new generation of packaging solutions which will be sustainable, customised, and green.


2022 - Trace elements in stomach oil of Scopoli's shearwater (Calonectris diomedea) from Linosa's colony [Articolo su rivista]
Cumbo, V.; Galluzzo, F. G.; Cammilleri, G.; Mascetti, A.; Lo Cascio, G.; Giangrosso, I. E.; Pulvirenti, A.; Seminara, S.; Ferrantelli, V.
abstract

Calonectris diomedea is a colonial Procellariiform breeding on Mediterranean islands. The stomach oil produced during chick rearing is a peculiar trait of this species. The composition of the stomach oil is likely to reflect the composition of the prey ingested and might reveal the contaminants uptake with prey becoming a possible tool for the marine pollution monitoring. We examined the concentration of 15 trace elements by ICP-MS and direct mercury analyser. The principal component analysis revealed a heterogeneous pattern of metal concentration, showing a significant separation between samples collected 20 and 70 days after hatching. The data obtained in this work give preliminary information on the feeding habits and breeding ecology of Linosa's colony of Scopoli's shearwater. The trace metals variability found suggest that the stomach oil may have a role as trophic markers to understand predator-prey relationships and to have evidence on the accumulation of pollutants in the latter.


2022 - Un nuovo biostimolante dai sottoprodotti della filiera ittica [Articolo su rivista]
Caser, M; Masino, F; Gervasio, S; Fassio, F; Ben Hassine, M; Benedetti, P; Caradonia, F; Cocchiara, S; Milc, J. A.; Caccialupi, G; Pulvirenti, A; Antonelli, A; Torri, L; Cevasco, R; Rovera, F; Scariot, V; Francia, E; Bertora, C; Ertani, A; Bertelli, C.
abstract

I sottoprodotti della pesca contengono proteine, grassi e amminoacidi, rendendoli così ottimi candidati per la produzione di biostimolanti. Il progetto FiSH (Fertilizzante idrolizzato Suolo e Habitat), al quale collaborano sei aziende agricole liguri, ha come obiettivo lo studio di un idrolizzato proteico fermentato derivato da scarti del pescato sulla crescita, resa e qualità di diverse colture floricole e orticole (Ranunculus asiaticus, Anemone x hybrida ‘Mistral Blu’, Solanum tuberosum ‘Quarantina’, Ocimum basilicum, Cucurbita pepo ‘Trombetta d’Albenga’, Allium sativum e Allium cepa). Durante il primo anno di progetto (2020-2021) sono stati testati in campo quattro concentrazioni (0,3 ml/L, 0,5 ml/L, 1 ml/L e 2 ml/L) del prototipo Fish n.7 in applicazione fogliare. Su patata è stata prevista anche un’applicazione su tubero preimpianto e valutato l’attacco di peronospora (Phytophthora infestans). I risultati ottenuti hanno evidenziato un’ampia variabilità di risposta. Tra le sei varietà di ranuncolo, ‘Elengance bianco’ ha mostrato una tendenza a produrre piante più rigogliose e fiori più grandi in risposta alla dose più elevata di Fish n.7, mentre la dose 0,5 ml/L ha indotto un maggiore sviluppo vegetativo e maggior produzione di rizomi nella varietà ‘Venere’. Su patata i risultati suggeriscono di evitare l’immersione del tubero, di mantenere i trattamenti rameici e di utilizzare concentrazioni < 1 ml/L. Su basilico e zucchina è stata osservata una maggiore produzione di biomassa nelle piante trattate alla concentrazione 2 ml/L. Nessun effetto è emerso in anemone, cipolla e aglio anche se, i dati delle ultime due specie sembrano indicare un beneficio fornito dalla dose 1 ml/L. Sulla base dei risultati, sono in corso valutazioni con otto nuovi prodotti.


2021 - Acrylamide assessment of wheat bread incorporating chia seeds (Salvia hispanica L.) by LC-LM/MS [Articolo su rivista]
Galluzzo, Francesco Giuseppe; Cammilleri, Gaetano; Pantano, Licia; Lo Cascio, Giovanni; Pulvirenti, Andrea; Macaluso, Andrea; Vella, Antonio; Ferrantelli, Vincenzo
abstract

We examined the acrylamide content in samples of wheat bread with chia seeds added at different concentrations (2%, 5%, 7%, 10%) and cooked at predefined conditions (20 min at 200°C) by a validated LC-MS/M method after QuEChERS extraction. The acrylamide contents of the bread samples with added chia seeds were compared with control wheat bread samples. The highest acrylamide values were found in bread with 5% chia seeds, showing a mean value of 156.5 ± 115.4 µg/kg, followed by bread with 10% chia seeds (150.2 ± 103.8 µg/kg). About 6% of the bread samples with added chia seeds reached acrylamide levels above the benchmark level set by the EU Regulation. No significant differences in acrylamide values were found between control samples and bread with different percentages of chia seeds (p > .05). The results obtained provide a first report on the possible contribution of chia to the increase of acrylamide formation in bread.


2021 - Characterization of chitosan-hydroxypropyl methylcellulose blend films enriched with nettle or sage leaf extract for active food packaging applications [Articolo su rivista]
Bigi, F.; Haghighi, H.; Siesler, H. W.; Licciardello, F.; Pulvirenti, A.
abstract

The incorporation of plant leaf extracts into biodegradable food packaging materials is a promising green approach to develop active films with antioxidant and antimicrobial activities. In this context, this study aimed to develop active films based on chitosan/hydroxypropyl methylcellulose blend (CS/HPMC) enriched with sage (SLE) and nettle (NLE) leaf extracts (7.5–15% w/w of biopolymer) to characterize their surface and cross-section morphology, optical, mechanical, water barrier, and antioxidant properties for food packaging applications. Scanning electron microscopy confirmed microstructural integrity and compatibility between CS and HPMC and incorporated leaf extracts. The successful incorporation of plant extracts was confirmed by Fourier transform infrared spectroscopy. Active films showed improvement in UV–Vis light barrier properties (p < 0.05) with opacity value lower than five. Addition of leaf extracts induced a slightly darker color by inducing a green and yellow shade. Addition of NLE increased the water solubility and water vapor permeability compared to the control film (p < 0.05). Total phenolic content and antioxidant activity were increased upon addition of leaf extracts. Overall, CS/HPMC films incorporated with SLE and NLE could be employed as a green alternative for partial substitution of synthetic plastics with antioxidant activity and to prolong the shelf-life of food products.


2021 - Domestic Use Simulation and Secondary Shelf Life Assessment of Industrial Pesto alla genovese [Articolo su rivista]
Nicosia, Carola; Fava, Patrizia; Pulvirenti, Andrea; Antonelli, Andrea; Licciardello, Fabio
abstract

The secondary shelf life (SSL) is defined as the time after package opening during which the food product retains a required level of quality. The SSL, indicated in labels as “best if used within x days after opening”, could lead to domestic food waste if not correctly evaluated. In this context, the SSL of two brands of industrial shelf-stable pesto products (with an indicated SSL of 5 days) was studied through a domestic use simulation performed in five households under two scenarios simulating real opening and storage conditions. The quality of pesto after opening was assessed through microbiological and sensory analyses, determination of instrumental colour parameters, pH and volatiles profiling. For both pesto sauces tested, a SSL 20 days was proven. Irrespective of the intensity of use (scenarios 1 and 2), the pesto was microbiologically stable: the maximum count for total aerobic mesophilic bacteria (TMB) observed during 20 days of storage was 9.64 1.7 102 CFU/g, starting from a commercially stable product. Colour parameters L* and DE did not change significantly during storage (p > 0.05), while the a* and BI values significantly changed (p < 0.05) during the first 5 days, and then stabilized during the rest of the household storage. Nevertheless, the slight colour modifications were not perceived by the sensory panel. Moreover, sensory assessors were not able to discern pesto samples stored for up to 20 days after first opening, from a just-opened reference sample, proving that the sensory appreciation of pesto was not influenced by the time after opening. The results of this study suggest the possibility to significantly extend or even omit the SSL indications for industrial pesto sauces. The objective assessment of SSL could have impressive practical outcomes both for the industry and the end user. The elongation of the SSL on the food label might increase food sustainability, thanks to the potential reduction of food wastes, thus giving added value to the commercial products. In addition, the end user could benefit the increase of the useful period for the food consumption after first opening, with significant domestic food waste reduction, reduced household stock turnover and consequent cost savings.


2021 - Effect of Time and Temperature on Physicochemical and Microbiological Properties of Sous Vide Chicken Breast Fillets [Articolo su rivista]
Haghighi, Hossein; Belmonte, Anna Maria; Masino, Francesca; Minelli, Giovanna; Lo Fiego, Domenico Pietro; Pulvirenti, Andrea
abstract

Temperature and time are two critical parameters in sous vide cooking which directly affect eating quality characteristics and food safety. This study aimed to evaluate physicochemical and microbiological properties of sous vide chicken breast fillets cooked at twelve different combinations of temperature (60, 70, and 80 C) and time (60, 90, 120, and 150 min). The results showed that cooking temperature played a major role in the moisture content, cooking loss, pH, a* color value, shear force, and thiobarbituric acid reactive substances (TBARS). Increasing cooking temperature caused an increase in cooking loss, lipid oxidation, TBARS, and pH, while moisture content was reduced (p < 0.05). Cooking time played a minor role and only moisture content, cooking loss, and a* color value were affected by this parameter (p < 0.05). Total mesophilic aerobic bacteria, Psychrotrophic bacteria, and Enterobacteriaceae were not detected during 21 days of storage at 4 C. Cooking at 60 C for 60 min showed the optimum combination of temperature and time for sous vide cooked chicken breast fillets. The result of this study could be interesting for catering, restaurants, ready-to-eat industries, and homes to select the optimum combination of temperature and time for improving the eating quality characteristics and ensuring microbiological safety.


2021 - Efficacy of an antifungal edible coating for the quality maintenance of Tarocco orange fruit during cold storage [Articolo su rivista]
Strano, Maria Concetta; Restuccia, Cristina; De Leo, Riccardo; Mangiameli, Solidea; Bedin, Elisa; Allegra, Maria; Quartieri, Andrea; Cirvilleri, Gabriella; Pulvirenti, Andrea
abstract

This study evaluated the effectiveness of a novel pectin-based edible coating (PBEC), alone or added with the antagonistic yeast Wickerhamomyces anomalus BS91 (PBECY), in reducing postharvest rots of Tarocco orange fruit. Their antifungal efficacy was assessed in vitro and in vivo, as well as their ability to preserve fruit quality under refrigerated conditions on three different cultivars, comparing a commercial wax amended with a fungicide. For all the treatments, the decay incidence, physiological disorders, weight loss and physical-chemical parameters were evaluated. The in vivo tests results indicated that PBECY enhanced the antifungal effect compared to PBEC, given the significant reduction (p ≤ 0.01) of Penicillium digitatum (PD) decay incidence (up to 90%). After 30 d of cold storage, both PBEC and PBECY reduced fruit decay (p≤ 0.05) in all cultivars, with values between 0 and 1.2%, relatively to the untreated control (up to 10%). Pectin-coated fruit showed a cultivar-dependent weight loss, with values comparable or higher to the other treatments, respectively for T. Lempso and T. Messina, whereas weigh loss in T. Sciara was higher only when coated with PBEC. Fruit physical and chemical parameters were preserved until 30 d of storage, thus demonstrating the potential of this edible coating for future industrial application.


2021 - Functional and Healthy Features of Conventional and Non-Conventional Sourdoughs [Articolo su rivista]
De Vero, Luciana; Iosca, Giovanna; Gullo, Maria; Pulvirenti, Andrea
abstract

Sourdough is a composite ecosystem largely characterized by yeasts and lactic acid bacteria which are the main players in the fermentation process. The specific strains involved are influenced by several factors including the chemical and enzyme composition of the flour and the sourdough production technology. For many decades the scientific community has explored the microbiological, biochemical, technological and nutritional potential of sourdoughs. Traditionally, sourdoughs have been used to improve the organoleptic properties, texture, digestibility, palatability, and safety of bread and other kinds of baked products. Recently, novel sourdough-based biotechnological applications have been proposed to meet the demand of consumers for healthier and more natural food and offer new inputs for the food industry. Many researchers have focused on the beneficial effects of specific enzymatic activities or compounds, such as exopolysaccharides, with both technological and functional roles. Additionally, many studies have explored the ability of sourdough lactic acid bacteria to produce antifungal compounds for use as bio-preservatives. This review provides an overview of the fundamental features of sourdoughs and their exploitation to develop high value-added products with beneficial microorganisms and/or their metabolites, which can positively impact human health.


2021 - Impact of low‐dose gaseous ozone treatment to reduce the growth of in vitro broth cultures of foodborne pathogenic/spoilage bacteria in a food storage cold chamber [Articolo su rivista]
Bigi, Francesco; Haghighi, Hossein; Quartieri, Andrea; De Leo, Riccardo; Pulvirenti, Andrea
abstract

Cold storage coupled with gaseous ozone represents a potential strategy to reduce or inhibit the presence of pathogenic and spoilage bacteria in a food storage cold chamber. This study aims to evaluate the impact of gaseous ozone treatment (0.05 ppm at exposure times of 30 and 60 min) on the bacterial contamination of internal surface and air in a cold chamber (3C) intended for food storage. The bacterial load of internal surfaces was reduced by 0.99 ± 0.24 and 1.35 ± 0.27 log after 30 and 60 min ozone treatment, respectively. Airborne bacterial load was reduced by 0.93 ± 0.24 log after 30 min ozone treatment and became non-detectable after 60 min. Gaseous ozone treatments (0.05 ppm at exposure times of 1, 2, 6, 24, 30, and 48 hr) of the cold chamber were investigated to evaluate the effectiveness of this technology against Escherichia coli, Listeria monocytogenes, Salmonella enterica Typhimurium, Campylobacter jejuni, and Pseudomonas fluorescens cultured in broth cultures. Ozone treatment was effective against C. jejuni since the population at the highest concentration of inoculum (3.34 log CFU/plate) was reduced by 2.23 log after 1 hr and it was completely undetectable after 2 hr. S. enterica and E. coli showed the highest resistance to short ozone treatment since 6 hr treatment did not show antibacterial activity whereas after 24 hr treatment around 2 log reduction was observed for both pathogens. Short ozone treatment did not affect L. monocytogenes viability. P. fluorescens showed high sensitivity to short treatments, with 0.75–1.32 log reductions after 1 hr and further 0.22–0.53 log reductions after 6 hr.


2021 - Multivariate exploratory data analysis by PCA of the combined effect of film-forming composition, drying conditions, and UV-C irradiation on the functional properties of films based on chitosan and pectin [Articolo su rivista]
Bigi, F.; Haghighi, H.; De Leo, R.; Ulrici, A.; Pulvirenti, A.
abstract

The technical features of biodegradable films depend on several manufacturing parameters. Exploiting these films on an industrial basis requires statistical models for the fast and effective evaluation of these parameters. This study analyses the impact of compositional and drying factors on the microstructural and functional properties of films based on chitosan and pectin through a multivariate approach. A total of 32 formulations were developed and the results were analysed through principal component analysis (PCA). Four formulations did not form films due to the photodegradation caused by prolonged UV-C irradiation. Films containing glycerol had a higher elasticity (1.5–39%) than those with propylene glycol or that were not plasticized (1.2–12%). Glycerol increased the thickness of pectin films (56–77 μm). Sodium trimethaphosphate (STMP) enhanced the UV mean absorbance of both pectin and chitosan films (0.7–1.7). In addition, STMP led to an increase of thickness (72–129 μm) and air permeability (38.6 × 103 to 9206.1 × 103 cm3/m2 × day × kPa) of the chitosan films, while CaCl2 led to a decrease of thickness (31–59 μm) and air permeability (23.9 × 103 to 46.8 × 103 cm3/m2 × day × kPa) of the pectin films.


2021 - Selection of Promising Exopolysaccharide-Producing Starter Cultures for Gluten-Free Sourdough [Abstract in Rivista]
Iosca, Giovanna; Febo, Camilla; Haghighi, Hossein; De Vero, Luciana; Gullo, Maria; Pulvirenti, Andrea
abstract

Celiac disease is an autoimmune enteropathy that affects the small intestine and is caused by the gliadin fraction of wheat gluten and other alcohol-soluble proteins (prolamines) of barley and rye in genetically predisposed subjects. Nowadays, the only recognized cure for this pathology is the use of gluten-free (GF) products. Although the market for GF products has grown in the last few years, their improvement is still a challenge for the food industry. With regard to GF bakery products, some of them can result in an unappetizing taste with a high fat content. These inconveniences can be minimized or solved by using sourdough fermentation. Sourdough is a stable culture of lactic acid bacteria (LAB) and yeast in a mixture of flour and water. In particular, GF sourdough represents a rich source of naturally occurring LAB and wild yeasts, which play important roles in food fermentation. This is mainly due to their potential in improving functional, technological, and probiotics properties, contributing to safe food with a better taste. Moreover, exopolysaccharide (EPS)-producing strains can also contribute to improving the sensory and rheological quality of the different GF products. This study aims to isolate and characterize the microbial pool from a GF sourdough made with rice flour and to evaluate the ability of selected LAB strains to produce exopolysaccharides (EPS). The yeast and LAB were isolated in selective agar media. A total of 220 isolates (110 yeasts and 110 bacteria) were obtained. They were taxonomically identified and characterized using both traditional and molecular approaches. Sequencing analysis of the D1/D2 domain of the 26S rRNA and the 16S rRNA genes, for yeasts and bacteria, respectively, revealed the occurrence of Saccharomyces cerevisiae as predominant yeasts and Lactiplantibacillus plantarum and Pediococcus pentosaceus as dominant LAB. Regarding the EPS production, 25 strains were able to produce dextran-like EPS with a maximum yield of 2 g/L. This study allowed the isolation and selection of promising EPS-producing starter cultures for the different types of flour sourdough, in particular, GF sourdough


2021 - Suitability Assessment of PLA Bottles for High-Pressure Processing of Apple Juice [Articolo su rivista]
Cubeddu, Arianna; Fava, Patrizia; Pulvirenti, Andrea; Haghighi, Hossein; Licciardello, Fabio
abstract


2021 - The cold chain and the COVID-19 pandemic: an unusual increase in histamine content in fish samples collected in Southern Italy during lockdown [Articolo su rivista]
Galluzzo, Francesco Giuseppe; Cammilleri, Gaetano; Cicero, Antonello; Pantano, Licia; Pulvirenti, Andrea; Macaluso, Andrea; Cicero, Nicola; Calabrese, Vittorio; Ferrantelli, Vincenzo
abstract

Objectives We analysed 900 samples of fresh (250) and processed (650) fish products collected in Sicily (Southern Italy) in 2020 during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic (hereafter: COVID-19). Materials and methods The samples were divided temporally based on five phases relating to the various restrictions imposed by the Italian government in this period. The validated method of ultra-high performance liquid chromatography (UHPLC) combined with a diode array detector (DAD) was then employed for the analysis. Results The samples collected during the Phase I lockdown period and after it had ended (Phase II) revealed significant increases in the mean histamine levels: 41.89±87.58 mg/kg -1 and 24.91±76.76 mg/kg -1, respectively. The 11 (1.3% of the total) fresh fish samples that were identified as being non-compliant with EC Reg. 2073/2005 were only found during these two periods. All the processed samples were always compliant. The histamine values decreased as the restrictions eased, achieving a mean value of 11.16±9.3 mgkg -1 (Phase III). Conclusions There was an increase in the incidence of fish samples that were non-compliant with EC Reg. 2073/2005 compared to previous surveillance data. These results provide a first report on the effect of lockdown measures on food safety and the cold chain. Our findings must cause food safety operators to intensify their controls over fresh fish products in such periods to safeguard consumer health. Further studies are required to evaluate whether the same trend would be observed with other food contaminants.


2021 - Yeasts and Lactic Acid Bacteria for Panettone Production: An Assessment of Candidate Strains [Articolo su rivista]
De Vero, Luciana; Iosca, Giovanna; La China, Salvatore; Licciardello, Fabio; Gullo, Maria; Pulvirenti, Andrea
abstract

The recovery of yeasts and lactic acid bacteria (LAB) involved in sourdough fermentation is the first step in the selection of starters with suitable technological aptitude and capable of producing desired aromas and/or aromatic precursors. In this work, two sourdoughs samples (MA and MB) and the derived doughs (samples A and B) were collected from a bakery during artisanal Panettone manufacture. Yeasts and bacteria were isolated at different fermentation steps on selective agar media. A total of 77 isolates were obtained and characterized. Representative strains of yeasts and LAB were identified by sequencing the D1/D2 domain of the 26S rRNA and the 16S rRNA genes, respectively. Moreover, the volatile organic compounds (VOCs) produced in the collected samples were detected and correlated to the species found in the same samples. The results highlighted the occurrence of Kazachstania humilis in both samples A and B, while Saccharomyces cerevisiae strains were detected only in samples B. Among LAB, Fructilactobacillus sanfranciscensis was the main species detected in both sourdoughs. Furthermore, strains belonging to the species Lactiplantibacillus plantarum, Furfurilactobacillus rossiae, Lactobacillus parabuchneri, Leuconostoc citreum, and Leuconostoc mesenteroides were assessed in the dough samples.


2020 - Antimicrobial peptide cocktail activity in minced turkey meat [Articolo su rivista]
Palman, Yael; De Leo, Riccardo; Pulvirenti, Andrea; Green, Stefan J.; Hayouka, Zvi
abstract

Meat products contain valuable nutrients that are important for human health and development but are also highly susceptible to colonization by microorganisms. This can lead to spoilage and serious foodborne illnesses. Natural antimicrobial peptides, produced by many organisms as part of their innate immune system to fight microbial infections, have great potential as food preservatives. In this study, we explored the effect of ternary antimicrobial random peptide mixtures (RPMs) on food spoilage bacteria in minced turkey meat. Amendment of RPMs to meat led to significant reductions in bacterial abundance in experimental tests, and RPMs worked synergistically with nitrite to reduce bacterial loads. Using high-throughput 16 S ribosomal RNA gene amplicon sequencing, we characterized the effect of RPMs and nitrite on meat microbial community structure before and during incubation under refrigerated conditions. Our findings reveal strong antimicrobial activity for RPMs against spoilage bacteria in meat, including Listeria monocytogenes and Pseudomonas putida. These results demonstrate the potential of RPMs as a safer preservative for reducing spoilage in meat and other food products.


2020 - Bioplastic Film From Black Soldier Fly Prepupae Proteins Used As Mulch: Preliminary Results [Articolo su rivista]
Setti, Leonardo; Francia, Enrico; Pulvirenti, Andrea; De Leo, Riccardo; Martinelli, Simone; Maistrello, Lara; Macavei, Laura Ioana; Montorsi, Monia; Barbi, Silvia; Ronga, Domenico
abstract

A protein‐based film (PBF), obtained from black soldier fly prepupae proteins, was assessed for its agronomic performance as mulch. PBF was investigated in a potting experiment and compared with Mater‐Bi (MB), polyethylene (PE) and bare soil. During the experiment, degraded surface area, weight and thickness of the film, water evaporated from the pot, and the soil microbiological content (SMC) were recorded. In addition, films were buried to assess their biodegradability and impact on SMC. During the mulching process, the PBF showed a significant degradation. In terms of evaporated water, the PBF performed similarly to MB and worse than PE. Regarding SMC, film of any nature caused an increase in the Clostridia spp. and a decrease of total mesophilic aerobic bacteria and fungi contents. When buried, only PBF recorded a faster biodegradability, showing a reduction of surface area, thickness and weight. PBF and MB highlighted a significant increase in contents of Clostridia spp., mesophilic aerobic bacteria and fungi. Our results reported, for the first time, the valorization of black soldier fly (BSF) prepupae proteins as a biodegradable film for mulching purposes. However, further study is needed to reduce the PBF biodegradability and allow it to be used for the most important mulched crops.


2020 - Characterization of bio-nanocomposite films based on gelatin/polyvinyl alcohol blend reinforced with bacterial cellulose nanowhiskers for food packaging applications [Articolo su rivista]
Haghighi, Hossein; Gullo, Maria; La China, Salvatore; Pfeifer, Frank; Siesler, Heinz Wilhelm; Licciardello, Fabio; Pulvirenti, Andrea
abstract

Bacterial cellulose nanowhiskers (BCNW) were synthesized from Komagataeibacter xylinus (strain K2G30; UMCC 2756) using sulfuric acid hydrolysis and incorporated into a gelatin-polyvinyl alcohol (GL/PVA) blend film matrix. The effect of BCNW content (1–10 wt% of biopolymer) on the microstructural, mechanical, optical, and water barrier properties of bio-nanocomposites was studied. Transmission electron microscopy showed that BCNW had a needle shape morphology with an average length of 600 nm and an average width of 30 nm. The crystallinity index of BCNW was 94.7% using X-ray diffraction. Scanning electron microscopy (SEM) illustrated good miscibility between GL/PVA blend film matrix and BCNW up to 7.5 wt%. Fourier-transform infrared spectroscopy in the attenuated total reflection mode showed molecular interactions between functional groups of the GL/PVA blend film matrix and BCNW. The incorporation of BCNW up to 7.5% into the GL/PVA blend reduced the water vapor transmission rate and water vapor permeability by about 22% and 14%, respectively, while tensile strength, elongation at break, and elastic modulus increased by about 21.5%, 41% and 19%, respectively (p <0.05). Films transparency was not affected by the addition of BCNW (p >0.05) suggesting that the BCNW were dispersed uniformly at the nanoscale. All films were colorless (ΔE*<2) with low opacity value (<2) comparable to synthetic plastics. Overall, the characterization of functional properties revealed that GL/ PVA blend film reinforced with BCNW could be used as an environmentally friendly packaging material to partially replace or reduce the use of current petroleum-based packaging materia


2020 - Combined Effects of Dewatering, Composting and Pelleting to Valorize and Delocalize Livestock Manure, Improving Agricultural Sustainability [Articolo su rivista]
Ronga, Domenico; Mantovi, Paolo; Pacchioli, Maria Teresa; Pulvirenti, Andrea; Bigi, Francesco; Allesina, Giulio; Pedrazzi, Simone; Tava, Aldo; Dal Prà, Aldo
abstract

An agronomic strategy to mitigate climate change impact can be the build-up of soil organic carbon. Among agronomic management approaches, the administration of organic fertilizers like livestock manure represents an eective strategy to increase soil organic carbon. However, livestock manure usually contains a high amount of water, reducing its sustainable delocalization and impacting on greenhouse gas emissions and nutrient leaching. Furthermore, the possible presence of weed seeds and harmful microorganisms could reduce the agronomic value of the manure. To overcome these issues, the combined eects of dewatering, composting and pelleting were investigated on livestock manure to produce sustainable organic fertilizers. Our results showed that composting and pelleting can represent a feasible and sustainable solution to reduce the potential risks related to the presence of weed seeds and harmful bacteria, concentrating nutrients and allowing a sustainable valorization and delocalization of the livestock manure. In addition, the processed manures were assessed as fertilizers in the growing medium (GM), displaying an increase in water retention and nutrient availability and a decrease of GM temperature and weed seed emergences. However, further study is needed to validate, both in open field and greenhouse productions, the eects of the proposed fertilizers in real cropping systems.


2020 - Development of a rapid and eco-friendly UHPLC analytical method for the detection of histamine in fish products [Articolo su rivista]
Cicero, A.; Galluzzo, F. G.; Cammilleri, G.; Pulvirenti, A.; Giangrosso, G.; Macaluso, A.; Vella, A.; Ferrantelli, V.
abstract

We developed, validated, and confirmed with proficiency tests a fast ultra-high-performance liquid chromatography with diode array detector (UHPLC-DAD) method to determine histamine in fish and fishery products. The proposed method consists of two successive solid–liquid extractions: one with a dilute solution of perchloric acid (6%) and the second only with water. The instrumental analysis with UHPLC provides a very fast run time (only 6 min) with a retention time of approximately 4 min, a limit of quantification (LOQ) of 7.2 mg kg−1, a limit of detection (LOD) of 2.2 mg kg−1, a recovery around 100%, a relative standard deviation (RSD%) between 0.5 and 1.4, and an r2 of calibration curve equal to 0.9995. The method detected optimal values of the validation parameters and required a limited number of reagents in comparison to other methods reported in the literature. Furthermore, the method could detect histamine in a very short time compared with other methods. This method, in addition to being validated, precise, specific, and accurate, avoids wasting time, money, and resources, and limits the use of organic solvents.


2020 - Development of antimicrobial films based on chitosan-polyvinyl alcohol blend enriched with ethyl lauroyl arginate (LAE) for food packaging applications [Articolo su rivista]
Haghighi, Hossein; KAMENI LEUGOUE, Serge; Pfeifer, Frank; Siesler, Heinz Wilhelm; Licciardello, Fabio; Fava, Patrizia; Pulvirenti, Andrea
abstract

The main aim of this study was to characterize microstructural, physical, optical, mechanical, water barrier and antimicrobial properties of chitosan-polyvinyl alcohol blend films (CS-PVA) enriched with ethyl lauroyl arginate (LAE) (1–10% w/v) for food packaging applications. The film microstructure was determined by scanning electron microscopy. Active films containing 10% LAE showed cracks on the surface with irregular shape in the cross-section indicating a weaker cohesion of the CS-PVA polymer blend at high LAE concentrations. The possible interaction of CS-PVA blend film with incorporated LAE was also investigated using Fourier-transform infrared (FT-IR) spectroscopy in the attenuated total reflection (ATR) mode. FT-IR/ATR spectra showed a low molecular interaction between the CS-PVA and LAE up to 2.5% while for films containing 5 and 10% LAE such interactions between the functional groups of the CS-PVA matrix and LAE have been detected. The active films were transparent and showed barrier properties against UV and visible light. The incorporation of LAE into the CS-PVA increased the thickness, water solubility, water vapor permeability, and the b* and ΔE* values, while it decreased mechanical properties and transparency (p < 0.05). Active films inhibited the growth of four major food bacterial pathogens including Campylobacter jejuni, Escherichia coli, Listeria monocytogenes and Salmonella typhimurium. Particularly, films containing 5 and 10% LAE were the most effective (p < 0.05). Overall, the characterization of functional properties revealed that CS-PVA blend film incorporated with LAE could be used as an environmentally friendly antimicrobial packaging material to extend the shelf life of food products.


2020 - Exploring the Microbial Community of Traditional Sourdoughs to Select Yeasts and Lactic Acid Bacteria [Articolo su rivista]
Iosca, Giovanna; Vero, Luciana De; Gullo, Maria; Licciardello, Fabio; Quartieri, Andrea; Pulvirenti, Andrea
abstract

Sourdoughs represent an awesome example of ecosystem in which yeasts and lactic acid bacteria (LAB) interact with each other, defining the characteristics of the final product in terms of composition, texture, taste and flavor. Therefore, the identification of dominant yeasts and LAB involved in the fermentation process can lead to the selection of starters with suitable fermentation aptitude and capable of producing desired aromas and/or aromatic precursors. In this work, two sourdoughs samples (A and B) for Panettone production were collected from an artisan bakery. Yeasts and bacteria were isolated at different fermentation steps on selective agar media. A total of 120 isolates were obtained and firstly characterized by conventional microbiological methods. Afterward, genomic DNA was extracted from the cultures, and (GTG)5-PCR fingerprinting analysis was carried out to reduce the redundance among the isolates. Representative yeasts and LAB strains, having a unique profile, were identified by sequencing the D1/D2 domain of the 26S rRNA and the 16S rRNA genes, respectively. The results highlighted the occurrence of Kazachstania humilis and Fructilactobacillus sanfranciscensis in both sourdoughs. Among LAB, also some other strains belonging to Lactobacillus genus were found. Moreover, Saccharomyces cerevisiae and Staphylococcus spp. strains were detected in sample B. In this study, a pool of yeasts and LAB strains for producing starter cultures with specific technological traits for sourdoughs production was obtained.


2020 - Genetic Improvement of wine yeasts for opposite adsorption activity of phenolics and ochratoxin A during red winemaking [Articolo su rivista]
Caridi, Andrea; Sidari, Rossana; Pulvirenti, Andrea; Blaiotta, Giuseppe
abstract

The aim of this research was to acquire new strains of Saccharomyces cerevisiae exhibiting opposite characteristics of cell wall adsorption: very high adsorption activity toward the ochratoxin A, very low adsorption activity toward the pigmented phenolic compounds contained in musts from black grapes. For this purpose, starting from 313 strains of Saccharomyces cerevisiae, 12 strains were pre-selected and used to obtain 27 intraspecific hybrids. Eleven crosses out of 27 were validated as hybrids; the best five hybrids were used in guided winemaking at four Calabrian wineries. The employed experimental protocol has allowed to select yeast strains for their different adsorption activity, improving the strains by spore clone selection and construction of intraspecific hybrids. These results suggest an efficacious way to improve the characteristics of interest in wine yeasts


2020 - High hydroxycinnamic acids contents in fennel honey produced in Southern Italy [Articolo su rivista]
Vella, Antonio; Cammilleri, Gaetano; Pulvirenti, Andrea; Galluzzo, Francesco; Randisi, Barbara; Giangrosso, Giuseppe; Macaluso, Andrea; Gennaro, Samuele; Ciaccio, Gabriele; Cicero, Nicola; Ferrantelli, Vincenzo
abstract

A total of 122 honey samples (Apis mellifera ssp. Ligustica) collected from Southern Italy were examined for floral identification by melissopalynological examination and for polyphenols detection by an LC-ESI-OrbitrapTM -MS/MS method. The melissopalynological examination confirmed all the samples examined as fennel (Foeniculum vulgare) unifloral variety. The analytical method carried out for polyphenols detection showed satisfactory linearity and recovery values, achieved during the validation of the method. Very high amounts of flavonols (kaempferol and quercetin) and hydroxycinnamic acids (caffeic acid, chlorogenic acid and ferulic acid), were found in all the samples examined. Among the hydroxycinnamic acids group, caffeic acid showed the highest mean contents (865.90 ± 67.07 µg/kg). The results of this work confirmed the high presence of phenolic acids with strong free radical-scavenging activity in fennel products such as honey, suggesting their use to reduce oxidative stress.


2020 - Histamine in fish products randomly collected in Southern Italy: A 6-year study [Articolo su rivista]
Cicero, A.; Cammilleri, G.; Galluzzo, F. G.; Calabrese, I.; Pulvirenti, A.; Giangrosso, G.; Cicero, N.; Cumbo, V.; Vella, A.; MacAluso, A.; Ferrantelli, V.
abstract

In total, 4,615 fresh and processed fish samples collected from 2010 to 2015 were analyzed for histamine by ultrahighperformance liquid chromatography with diode array detection. Histamine levels were detected in 352 (7.6%) samples, with a maximum of 4,110 mg kg1 and mean values of 908.9 6 1,226.79 and 344.01 6 451.18 mg kg1 for fresh and processed fish samples, respectively. No histamine levels were found in canned tuna and smoked fish samples in contrast to most of the data reported in the literature. A low percentage (2.79%) of noncompliant samples was found. The highest mean values were found during 2011 and 2015 for fresh and processed fish samples, respectively, showing a significant (P , 0.05) difference between the sampling years. The histamine contents found in fresh fish samples were significantly higher (P , 0.05) than those of processed samples. Most of the positive samples came from street vendors, suggesting the need to improve inspection measures in these commercial categories to ensure fish product safety.


2020 - Recent advances on chitosan-based films for sustainable food packaging applications [Articolo su rivista]
Haghighi, Hossein; Licciardello, Fabio; Fava, Patrizia; Siesler, Heinz Wilhelm; Pulvirenti, Andrea
abstract

The recent sharp increase of sensitivity towards environmental issues arising from plastic packaging has boosted interest towards alternative sustainable packaging materials. This new trend promotes the industrial exploitation of knowledge on chitosan-based films. Chitosan has been extensively investigated and used due to its unique biological and functional properties. However, inherent drawbacks including low mechanical properties and high sensitivity to humidity represent major limitations to its industrial applications, including food packaging. In the present study, the scientific literature of the last five years has been extensively reviewed (source: Web of Science) addressing chitosan-based films for their potential application in the food packaging industry. The contribution summarizes the various strategies adopted to overcome inherent drawbacks and improve the properties of chitosan-based films, with special regards for blending with natural and synthetic biopolymers.


2020 - Residues of 165 pesticides in citrus fruits using LC-MS/MS: a study of the pesticides distribution from the peel to the pulp [Articolo su rivista]
Calvaruso, Enza; Cammilleri, Gaetano; Pulvirenti, Andrea; Lo Dico, Gianluigi Maria; Lo Cascio, Giovanni; Giaccone, Vita; Vitale Badaco, Valeria; Ciprì, Valentina; Alessandra, Mobilia Maria; Vella, Antonio; Macaluso, Andrea; Di Bella, Calogero; Ferrantelli, Vincenzo
abstract

Abstract A sensitive LC–ESI-MS/MS method was developed for the determination of 165 pesticides in 50 citrus fruit samples collected in Sicily. Moreover, an evaluation of pesticides levels in the citrus layers (peel, albedo, and pulp) was carried out. The method presented acceptable trueness, precision, and linearity with LOQ of 5 μg/kg. The results obtained showed a high frequency of fungicides class pesticides in all the citrus samples examined (>95%) with the highest concentrations in the peel (4468 µg/Kg). A significant difference of concentrations was found between the layers of the citrus fruits analysed (p < 0.05). In particular, the peel and albedo present higher pesticides significantly higher than the pulp. Our findings confirming the widespread use of these substances in citrus cultivation and suggesting the importance of pesticides analysis in all the citrus fruit layers separately, considering the different interactions between the physicochemical characteristics of the matrices and the pesticides.


2020 - Seasonal trend of Anisakidae infestation in South Mediterranean bluefish [Articolo su rivista]
Cammilleri, Gaetano; Pulvirenti, Andrea; Costa, Antonella; Graci, Stefania; Collura, Rosaria; Buscemi, Maria Drussilla; Sciortino, Sonia; Vitale Badaco, Valeria; Vazzana, Mirella; Brunone, Mariagrazia; Vella, Antonio; Di Bella, Calogero; Ferrantelli, Vincenzo
abstract

A total of 1104 fish samples from markets of Sicily were analysed for the detection and species identification of Anisakidae nematodes. The preliminary analysis of the fish samples showed the presence of 2459 larvae. All the fish species revealed different prevalence of infestation, with a maximum of 100% for Lepidopus caudatus and a minimum of 4.5% in Sardina pilchardus. The 80% of the larvae examined by PCR-RFLP analysis belonged to Anisakis pegreffii species. The seasonal infestation trend of Anisakis was evaluated in all the fish sample examined. The results of the seasonal infestation trend showed a marked connection with the ecological aspects of the fish species examined. As far as we know, this work report for the first time important ecological aspects of Lepidopus caudatus specimens of South Mediterranean. This work could be useful to plan a seasonal fishing strategy aimed at reducing the health risks related to Anisakis.


2020 - Selection of Wine Saccharomyces cerevisiae Strains and Their Screening for the Adsorption Activity of Pigments, Phenolics and Ochratoxin A [Articolo su rivista]
Pulvirenti, Andrea; De Vero, Luciana; Blaiotta, Giuseppe; Sidari, Rossana; Iosca, Giovanna; Gullo, Maria; Caridi, Andrea
abstract

Ochratoxin A is a dangerous mycotoxin present in wines and is considered the principal safety hazard in the winemaking process. Several authors have investigated the ochratoxin A adsorption ability of Saccharomyces cerevisiae yeasts, and specifically selected strains for this desired trait. In the present work, a huge selection of wine yeasts was done starting from Portuguese, Spanish and Italian fermenting musts of dierent cultivars. Firstly, 150 isolates were collected, and 99 non-redundant S. cerevisiae strains were identified. Then, the strains were screened following a multi-step approach in order to select those having primary oenological traits, mainly (a) good fermentation performance, (b) low production of H2S and (c) low production of acetic acid. The preselected strains were further investigated for their adsorption activity of pigments, phenolic compounds and ochratoxin A. Finally, 10 strains showed the desired features. The goal of this work was to select the strains capable of absorbing ochratoxin A but not pigments and phenolic compounds in order to improve and valorise both the quality and safety of red wines. The selected strains are considered good candidates for wine starters, moreover, they can be exploited to obtain a further enhancement of the specific adsorption/non-adsorption activity by applying a yeast breeding approach.


2020 - Toxic mineral elements in Mytilus galloprovincialis from Sicilian coasts (Southern Italy) [Articolo su rivista]
Cammilleri, Gaetano; Galluzzo, Paola; Pulvirenti, Andrea; Giangrosso, Innocenzo Ezio; Lo Dico, Gianluigi Maria; Montana, Giovanna; Lampiasi, Nadia; Mobilia, Maria Alessandra; Lastra, Antonio; Vazzana, Mirella; Vella, Antonio; La Placa, Pietro; Macaluso, Andrea; Ferrantelli, Vincenzo
abstract

We assessed the relationship between V, Cr, Mn, Hg, As, Cd, Sn, Sb and Pb concentrations in Mytilus galloprovincialis samples from the coasts of Sicily and the expression of metallothioneins. Toxic mineral elements assessment was carried out by A.A. Spectrometry and ICP-MS. The metallothioneins expression was performed by q-PCR method. Low metals' levels were found in the mussel samples examined, in comparison with what was reported in literature. The highest mean values of toxic mineral elements were found in Gela (Cr 0.178 ± 0.03 mg/Kg, Mn 4.325 ± 0.012 mg/Kg, As 3.706 ± 0.009 mg/Kg, Sn 0.148 ± 0.014 mg/Kg, Sb 0.009 ± 0.004 mg/Kg e Pb 0.364 ± 0.01 mg/Kg). Significant levels of Hg were found in samples from Catania (0.014 ± 0.005 mg/Kg). Only vanadium and lead concentrations showed significant differences between sampling areas (p < 0.05). Molecular analysis verified a basal expression of Mt1 and the absence of over-expression of Mt2, confirming the low mineral's concentrations found in the samples examined.


2020 - Validation of a Commercial Loop-Mediated Isothermal Amplification (LAMP) Assay for the Rapid Detection of Anisakis spp. DNA in Processed Fish Products [Articolo su rivista]
Cammilleri, Gaetano; Ferrantelli, Vincenzo; Pulvirenti, Andrea; Drago, Chiara; Stampone, Giuseppe; Del Rocio Quintero Macias, Gema; Drago, Sandro; Arcoleo, Giuseppe; Costa, Antonella; Geraci, Francesco; Di Bella, Calogero
abstract

Parasites belonging to the Anisakis genera are organisms of interest for human health because they are responsible for the Anisakiasis zoonosis, caused by the ingestion of raw or undercooked fish. Furthermore, several authors have reported this parasite to be a relevant inducer of acute or chronic allergic diseases. In this work, a rapid commercial system based on Loop-Mediated Isothermal Amplification (LAMP) was optimised and validated for the sensitive and rapid detection of Anisakis spp. DNA in processed fish products. The specificity and sensitivity of the LAMP assay for processed fish samples experimentally infected with Anisakis spp. larvae and DNA were determined. The LAMP system proposed in this study was able to give positive amplification for all the processed fish samples artificially contaminated with Anisakis spp., giving sensitivity values equal to 100%. Specificity tests provided no amplification for the Contracaecum, Pseudoterranova, or Hysterothylacium genera and uninfected samples. The limit of detection (LOD) of the LAMP assay proposed was 102 times lower than the real-time PCR method compared. To the best of our knowledge, this is the first report regarding the application of the LAMP assay for the detection of Anisakis spp. in processed fish products. The results obtained indicate that the LAMP assay validated in this work could be a reliable, easy-to-use, and convenient tool for the rapid detection of Anisakis DNA in fish product inspection.


2019 - Aflatoxin M1 in cow, sheep, and donkey milk produced in Sicily, Southern Italy [Articolo su rivista]
Cammilleri, Gaetano; Graci, Stefania; Collura, Rosaria; Buscemi, Maria Drussilla; Vella, Antonio; Macaluso, Andrea; Giaccone, Vita; Giangrosso, Giuseppe; Cicero, Antonello; Lo Dico, Gianluigi Maria; Pulvirenti, Andrea; Cicero, Nicola; Ferrantelli, Vincenzo
abstract

Samples (n = 485) of raw (n = 394) or heat-treated (n = 91) milk of three different species (cow, n = 170; sheep, n = 133; donkey, n = 84), collected 2013–2016 in Western Sicily (Southern Italy), were analyzed for aflatoxin M1 (AFM1) by enzyme-linked immunosorbent assay (ELISA). Positive ELISA results were further analyzed by HPLC with fluorescence detection. Both methods had a detection limit for AFM1 in milk of 7 ng kg−1. ELISA yielded 12.9 and 5% positives in cows and sheep milk, respectively, all samples of donkey milk were negative. Levels of AFM1 were in most cases at 0.007–< 0.05 μg kg−1, only two samples (sheep milk) slightly exceeded the European Union maximum level of 0.05 μg kg−1. Only 6% of the samples were positive for AFM1 in a concentration range of 0.008–0.15 μg kg−1. Only milk samples collected directly from farms were positive. Overall, the levels were much lower than previously reported for Southern Italy cow and sheep milk samples purchased in retail stores. The results of this work indicate a continuous improvement of the feeding techniques on dairy farms of Southern Italy, which is essential to ensure consumers’ protection.


2019 - Comparative analysis of blend and bilayer films based on chitosan and gelatin enriched with LAE (lauroyl arginate ethyl) with antimicrobial activity for food packaging applications [Articolo su rivista]
Haghighi, Hossein; De Leo, Riccardo; Bedin, Elisa; Pfeifer, Frank; Siesler, Heinz Wilhelm; Pulvirenti, Andrea
abstract

Blend and bilayer bio-based active films were developed by solvent casting technique, using chitosan (CS) and gelatin (GL) as biopolymers, glycerol as a plasticizer and lauroyl arginate ethyl (LAE) as an antimicrobial compound. Blend films had higher tensile strength and elastic modulus and lower water vapor permeability than bilayer films (p < 0.05). Bilayer films demonstrated as effective barriers against UV light and showed lower transparency values (p < 0.05). FT-IR spectra indicated that interactions existed between CS and GL due to electrostatic interactions and hydrogen bond formation. However, the addition of LAE did not interfere in the network structure. Active films incorporated with LAE (0.1%, v/v) inhibited the growth of Listeria monocytogenes, Escherichia coli, Salmonella typhimurium and Campylobacter jejuni. This study highlighted the development of blend and bilayer bio-based active films based on CS and GL enriched with LAE for food packaging applications with improved physical, mechanical, barrier and antimicrobial properties.o


2019 - Comprehensive characterization of active chitosan-gelatin blend films enriched with different essential oils [Articolo su rivista]
Haghighi, Hossein; Biard, Syméon; Bigi, Francesco; De Leo, Riccardo; Bedin, Elisa; Pfeifer, Frank; Siesler, Heinz Wilhelm; Licciardello, Fabio; Pulvirenti, Andrea
abstract

Natural extracts and plant essential oils (EOs) have long been recognized as valid alternatives to synthetic food additives owing to their proved wide-spectrum antimicrobial capacity. The main aim of this study was to characterize the physical, mechanical, water barrier, microstructural and antimicrobial properties of chitosan-gelatin blend films enriched with cinnamon, citronella, pink clove, nutmeg and thyme EOs. The film microstructure determined by scanning electron microscopy, showed that all active films had heterogeneous surface: in particular, films including cinnamon, nutmeg and thyme EOs showed remarkable pores on the surface. The possible interaction of chitosan-gelatin blend film with incorporated EOs was investigated using Fourier-transform infrared (FT-IR) spectroscopy. Presence of new bands and changes in the FT-IR spectra confirmed intermolecular interactions between the chitosan-gelatin matrix and the EOs. The antimicrobial activity of films was determined using the disk diffusion assay. Active films inhibited the growth of four major food bacterial pathogens including Campylobacter jejuni, Escherichia coli, Listeria monocytogenes and Salmonella typhimurium and, among the tested EOs, thyme was the most effective (p &lt; 0.05). The active films can be considered as effective barriers against UV light. The incorporation of EOs to the chitosan-gelatin film increased thickness, moisture content, water vapor permeability, b* and ΔE* values (p &lt; 0.05) while it decreased L* value, light transparency and opacity (p &lt; 0.05). Overall, the characterization of functional properties revealed that chitosan-gelatin films incorporated with EOs could be used as environmentally friendly active food packaging with antimicrobial properties and potential to extend the shelf-life of food products.


2019 - Effects of solid and liquid digestate for hydroponic baby leaf lettuce (Lactuca sativa L.) cultivation [Articolo su rivista]
Ronga, Domenico; Setti, Leonardo; Salvarani, Chiara; De Leo, Riccardo; Bedin, Elisa; Pulvirenti, Andrea; Milc, Justyna; Pecchioni, Nicola; Francia, Enrico
abstract

Digestate was evaluated as an alternative and sustainable growing medium and nutrient solution in the hydroponic cultivation of baby leaf lettuce (Lactuca sativa L.). Nine hydroponic combinations of substrate and fertilization (agriperlite + standard solution, agriperlite + liquid digestate, solid digestate + standard solution, solid digestate + liquid digestate, soil + standard solution, peat moss + standard solution; peat moss + liquid digestate, pelleted digestate + standard solution and pelleted digestate + liquid digestate) were tested and compared for the cultivation of baby leaf lettuce, in three different experiments. During the crop cycles, yield as other agronomical and microbiological parameters were investigated. The combination of agriperlite + liquid digestate, solid digestate + standard solution and pelleted digestate + standard solution enhanced plant growth by affecting the root, the shoot, the total dry weight and SPAD parameters, in the all investigated experiments (+32%, +40%, +29%, +17% respectively). Based on the obtained results, digestate represents a sustainable and alternative growing media or nutrient solution for the production of baby leaf lettuce cultivated in hydroponic system.


2019 - Features of chitosan-gelatin films loaded with plant extracts [Articolo su rivista]
Haghighi, H.; Siesler, H. W.; Licciardello, F.; Pulvirenti, A.
abstract

The main aim of this study was to characterize the physical, optical and antimicrobial properties of chitosan-gelatin blend films enriched with cinnamon, nutmeg and thyme essential oils. Microstructure determined by scanning electron microscopy showed that active films had heterogeneous surface. The presence of new bands and changes in the FTIR spectra confirmed intermolecular interactions between the chitosan-gelatin matrix and essential oils. The active films proved as effective barriers against UV light and inhibited the growth of four major food bacterial pathogens, showing potential as environmentally friendly antimicrobial packaging material.


2019 - Land Snails as a Valuable Source of Fatty Acids: A Multivariate Statistical Approach [Articolo su rivista]
Galluzzo, Francesco Giuseppe; Cammilleri, Gaetano; Ulrici, Alessandro; Calvini, Rosalba; Pulvirenti, Andrea; Lo Cascio, Giovanni; Macaluso, Andrea; Vella, Antonio; Cicero, Nicola; Amato, Andtonella; Ferrantelli, Vincenzo
abstract

The fatty acid (FA) profile of wild Theba pisana, Cornu aspersum, and Eobania vermiculata land snail samples, collected in Sicily (Southern Italy), before and after heat treatment at +100 °C were examined by gas chromatography with a flame ionization detector (GC‐FID). The results show a higher content of polyunsaturated fatty acids (PUFAs) in all of the examined raw snails samples, representing up to 48.10% of the total fatty acids contents, followed by monounsaturated fatty acids (MUFAs). The thermal processing of the snail samples examined determined an overall reduction of PUFA levels (8.13%, 7.75%, and 4.62% for T. pisana, C. aspersum and E. vermiculata samples, respectively) and a species‐specific variation of saturated fatty acid (SFA) contents. Oleic acid remained the most abundant FA of all of the snails species examined, accounting for up to 29.95% of the total FA content. A relevant decrease of ɷ3/ɷ6 ratio was found only for T. pisana samples. The principal component analysis (PCA) showed a separation of the snail samples in terms of species and heat treatment. The results of this work suggest land snails as a valuable source of MUFA and PUFA contents and boiling as appropriate treatment, according to the maintenance of healthy properties.


2019 - MICROBIOLOGICAL ASSESSMENT OF LAND SNAIL’S CAVIAR PRODUCED IN SICILY, SOUTHERN ITALY: A FIRST REPORT Microbial Diversity 2019 [Abstract in Atti di Convegno]
Sciortino, S.; Pulvirenti, A.; Cammilleri, G.; Cardamone, C.; Giangrosso, G.; Fiorista, C.; Vella, Arianna; Ferrantelli, V.
abstract

Terrestrial gastropods, commonly named snails, constitute a niche food product, traditionally appreciated by Italian consumers. Recently, attention has also been paid for land snails’ caviar, given the role they are playing in luxury gastronomy. The production of Sicilian snail caviar is constantly expanding, it is also exported to Spain, France, Portugal and Belgium. Sicily's climate facilitates its growth. Requested even in Russia, Switzerland and Dubai by starred chefs and in niche markets, caviar is currently sold at 1800 euros per kg, arousing curiosity and interest. However, as far we know, no toxicological and microbiological data are available in order to ensure the consumer health. This work aimed at give the first report on microbiological assessment of land snails’ caviar produced by Cornu aspersum species in Sicily (Southern Italy).All the sample pools examined were negative to the most tested microbes except to the Total mesophilic count, coliform, yeasts and moulds. The hygienic condition did not have a significant change after 10 and 20 days except for yeasts and moulds, that suffer a proportional increase of 1 log UFC/g. Regarding the pH and the free water, the sample pools showed overlapping values in the three analysis times.The results of microbiological analysis revealed good hygienic conditions of the product examined, showing an elevated shelf-life until 20 days of storage. The present work reported for the first time the health aspects of this innovative product, revealing the lack of necessity to the use of preservative additives (such as ascorbic acid etc.), that could compromise its organoleptic properties.


2019 - Mercury Detection in Benthic and Pelagic Fish Collected from Western Sicily (Southern Italy) [Articolo su rivista]
Cammilleri, Gaetano; Galluzzo, Francesco Giuseppe; Fazio, Francesco; Pulvirenti, Andrea; Vella, Antonio; Lo Dico, Gianluigi Maria; Macaluso, Andrea; Ciaccio, Gabriele; Ferrantelli, Vincenzo
abstract

In highly polluted water, fish can accumulate mercury up to a concentration of 10 mgKg-1. This has occurred on the eastern coasts of Sicily (Southern Italy), probably due to the intense industrial activity of this area. However, little is known about Hg accumulation in fish of the western Sicilian coasts. In this work, we examined the Hg accumulation of 108 fish samples belonging to 14 species collected from western Sicilian coasts using a direct mercury analyzer. The samples showed a mean mercury concentration of 0.165 ± 0.22 mg kg-1 with a maximum in Lepidopus caudatus (1.72 mgKg-1), exceeding the limits provided by EC Reg. 1881/2006. The lowest Hg levels were found in Sparus aurata samples (0.001 mgKg-1). A significant difference was found between the fish species examined (p &lt; 0.05). The comparison between benthic and pelagic species did not show statistical differences (p &lt; 0.05). Fish food constitutes the main route of Hg uptake for humans. Only four of the 130 samples examined reached a mercury concentration over the European limits. The comparative analysis of Hg pollution for benthic and pelagic species did not confirm a different trend in metal contamination.


2019 - Multivariate statistical analysis of the polyphenols content for the discrimination of honey produced in Sicily (Southern Italy) [Articolo su rivista]
Lo Dico, Gianluigi Maria; Ulrici, Alessandro; Pulvirenti, Andrea; Cammilleri, Gaetano; Macaluso, Andrea; Vella, Antonio; Giaccone, Vita; Lo Cascio, Giovanni; Graci, Stefania; Scuto, Maria; Trovato Salinaro, Angela; Calabrese, Vittorio; Lo Dico, Rea; Ferrantelli, Vincenzo
abstract

The polyphenols content of 105 honey samples produced by black honeybees (Apis Mellifera ssp. Sicula) and common honeybees (Apis mellifera ssp. Ligustica) from Western Sicily (Southern Italy) was examined using TurboFlow™ liquid chromatography Orbitrap™ high-resolution mass spectrometry. The results showed very high kaempferol and quercetin contents, with average values higher than what was reported in literature (3967.9±2184.16 and 2206.1±1666.4μg kg⁠−1 for kaempferol and quercetin, respectively). The honey samples produced by Apis Mellifera ssp. Sicula subspecies showed polyphenols content up to two times higher than Apis mellifera ssp. Ligustica. The Principal Component Analysis (PCA) model calculated on the polyphenols content showed a satisfactory separation of the honey samples in terms of honeybee subspecies and production area. The model proposed in this work shows the possibility to safeguard the authenticity of the honey produced in the various geographic areas of Sicily.


2019 - Preparation and characterization of active chitosan/bacterial cellulose nano-whisker composite film enriched with lauroyl arginate ethyl for food packaging applications - Microbial Diversity 2019 [Abstract in Atti di Convegno]
Haghighi, Hossein; LA CHINA, Salvatore; Gullo, Maria; Pulvirenti, Andrea
abstract

Acetic acid bacteria are recognized for their wide range of industrial applications. Recently, applying bacterial cellulose (BC) from acetic acid bacteria as a packaging material has received considerable attention from food packaging industries. This is mainly due to its unique properties such as biocompatibility, biodegradability, water holding capacity and remarkable mechanical properties (1). Active packaging is intended to extend the shelf-life of food products and assure their safety and quality inside the packaging systems. In this context, lauroyl arginate ethyl (LAE) is considered as one of the most effective antimicrobial substances among novel food additives which is also classified as GRAS (generally recognized as safe) by the Food and Drug Administration and the European Food Safety Agency (2). Thus, the main aim of this study was to characterize the microstructural, physical, mechanical and optical properties of chitosan/bacterial cellulose nano-whisker (BCNW) composite film enriched with LAE for potential application as active food packaging. Moreover, antimicrobial activity of this film against four common food bacterial pathogens including Campylobacter jejuni, Escherichia coli, Listeria monocytogenes and Salmonella typhimurium, was investigated.


2019 - Preservation, Characterization and Exploitation of Microbial Biodiversity: The Perspective of the Italian Network of Culture Collections [Articolo su rivista]
De Vero, Luciana; Boniotti, Maria Beatrice; Budroni, Marilena; Buzzini, Pietro; Cassanelli, Stefano; Comunian, Roberta; Gullo, Maria; Logrieco, Antonio F.; Mannazzu, Ilaria; Musumeci, Rosario; Perugini, Iolanda; Perrone, Giancarlo; Pulvirenti, Andrea; Romano, Paolo; Turchetti, Benedetta; Varese, Giovanna Cristina
abstract

Microorganisms represent most of the biodiversity of living organisms in every ecological habitat. They have profound effects on the functioning of any ecosystem, and therefore on the health of our planet and of human beings. Moreover, microorganisms are the main protagonists in food, medical and biotech industries, and have several environmental applications. Accordingly, the characterization and preservation of microbial biodiversity are essential not only for the maintenance of natural ecosystems but also for research purposes and biotechnological exploitation. In this context, culture collections (CCs) and microbial biological resource centres (mBRCs) are crucial for the safeguarding and circulation of biological resources, as well as for the progress of life sciences. This review deals with the expertise and services of CCs, in particular concerning preservation and characterization of microbial resources, by pointing to the advanced approaches applied to investigate a huge reservoir of microorganisms. Data sharing and web services as well as the tight interconnection between CCs and the biotechnological industry are highlighted. In addition, guidelines and regulations related to quality management systems (QMSs), biosafety and biosecurity issues are discussed according to the perspectives of CCs and mBRCs.


2019 - Tetracycline Residues in Bovine Muscle and Liver Samples from Sicily (Southern Italy) by LC-MS/MS Method: A Six-Year Study [Articolo su rivista]
Cammilleri, Gaetano; Pulvirenti, Andrea; Vella, Antonio; Macaluso, Andrea; Lo Dico, Gianluigi; Giaccone, Vita; Giordano, Vincenzo; Vinciguerra, Massimo; Cicero, Nicola; Cicero, Antonello; Giangrosso, Giuseppe; Vullo, Stefano; Ferrantelli, Vincenzo
abstract

We examined a total of 369 bovine liver and muscle samples for the detection of oxytetracycline (OTC), tetracycline (TC), chlortetracycline (CTC), and doxycycline (DOX) residues by implementation and validation of a LC-MS/MS method. The method showed good recovery values between 86% and 92% at three levels of concentrations. The linearity tests revealed r2 > 0.996 for all the tetracyclines examined. Furthermore, the Youden test revealed that the method was robust. Only 14.4% of the samples showed OTC and TC residues in a concentration range of 10.4–40.2 g kg1. No CTC and DOX residues were found in all the samples analyzed. Liver samples showed the highest average values (31.5 20.6 and 21.8 18.9 for OTC and TC, respectively). The results showed a low incidence of TCs in all the samples examined, in comparison with other studies reported in the literature. A significant decrease in TC residues frequency was found from 2013 (p < 0.05). This work reports for the first time epidemiological data on the presence of TC residues in liver and muscle samples of cattle farmed in Sicily (Southern Italy). The very low incidence of TC residues indicates a continuous improvement in farming techniques in Southern Italy, which is essential to ensure consumers’ protection.


2019 - Use of black soldier fly (Hermetia illucens (L.), Diptera: Stratiomyidae) larvae processing residue in peat-based growing media [Articolo su rivista]
Setti, Leonardo; Francia, Enrico; Pulvirenti, Andrea; Gigliano, Silvia; Zaccardelli, Massimo; Pane, Catello; Caradonia, Federica; Bortolini, Sara; Maistrello, Lara; Ronga, Domenico
abstract

The Black Soldier Fly (Hermetia illucens(L.), Diptera: Stratiomyidae) is an insect whose larvae thrive onagro-industrial by-products. This study reports the first use of black soldier fly larvae processing residue(BSPR) as an innovative ingredient for growing media. BSPR was characterized and evaluated to partiallyreplace commercial peat (CP) in the production of potted plants. Chemical and microbiological analysisshowed the suitability of BSPR for soilless production. Hence, six growing media mixtures (CP 100%+ slow acting synthetic solid fertilizer, CP 90% + BSPR 10%, CP 80% + BSPR 20%, CP 70% + BSPR 30%, CP60% + BSPR 40% and CP 100% without fertilizer) were assessed for the production of baby leaf lettuce,basil and tomato potted plants. Using BSPR in a proportion up to 20%, all investigated crops showed val-ues significantly greater than or comparable to those obtained using CP 100% + slow acting synthetic solidfertilizer. In general, BSPR used in a proportion up to 20% increased the crop growth of baby leaf lettuce,basil and tomato, recording a high total dry weight (+31%, compared to the total average) and the mea-sured leaf parameters (+39% of leaf area, +14% of leaf number), without showing abiotic stresses. Thisstudy indicates that BSPR used in a proportion up to 20% might be a valid approach for soilless productionof potted baby leaf lettuce, basil and tomato plants.


2018 - A LC-HRMS After QuEChERS Cleanup Method for the Rapid Determination of Dye Residues in Fish Products [Articolo su rivista]
Giaccone, Vita; Cammilleri, Gaetano; Macaluso, Andrea; Cicero, Nicola; Pulvirenti, Andrea; Vella, Antonio; Ferrantelli, Vincenzo
abstract

The reliability of a rapid LC-HRMS method was studied in order to find a sensitive and accurate, simple, cheap, and rapid method for screening and quantitative determination of malachite green (MG), leucomalachite green (LMG), brilliant green (BG), crystal violet (CV), and leucocrystal violet (LCV) in fish muscle. All the analytes were extracted using a mixture of acetonitrile and citrate buffer, while the cleanup phase was carried out by Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method. All the data analyzed were acquired using both full MS and dd-MS2 modes. Good specificity, precision (relative standard deviation <11% for each sample tested), recovery (10–60%), decision limit (CCα between 0.55 and 0.62 μg kg−1), detection capabilities (CCβ between 0.65 and 0.78 μg kg−1), and ruggedness were achieved for the reliability of the method. Satisfactory results were obtained during the linearity test in the range of 0.10–2 ng mL−1 (r2 > 0.999). Best recoveries were obtained by the QuEChERS cleanup method for all the analytes examined, presenting values between 70 and 104%. The application of 70,000 FWHM mass resolution and narrow mass windows significantly improved the selectivity of the method, leading to simultaneous screening and quantification of dye residues in comparison to other methods proposed in literature. The optimized method proposed in this work enables a simple and fast preparation; it offers exceptional sensitivity and selectivity and maximizes efficiency and reproducibility with a low consumption of reagents. Finally, the present method was successfully employed to detect dye residues in 73 fish samples, as provided for the national residue control plan.


2018 - Application of pectin-alginate and pectin-alginate-laurolyl arginate ethyl coatings to eliminate Salmonella enteritidis cross contamination in egg shells [Articolo su rivista]
DE LEO, Riccardo; Quartieri, Andrea; Haghighi, Hossein; Gigliano, Silvia; Bedin, Elisa; Pulvirenti, Andrea
abstract

This study highlights the potential application of pectin‐alginate blend (PA) and pectin‐alginate‐LAE blend (PAL) coatings to eliminate Salmonella enteriditis 10,118 cross‐contamination without changing the shelf‐life of fresh eggs and their physico‐chemical properties during storage at 7 °C for 42 days. Egg shells were dipped in a solution of Salmonella enteritidis 10,118 with a concentration of 7 x 106 cfu/ml to assess Salmonella cross‐contamination. PA and PAL coatings did not have a significant effect on shelf‐life based on physico‐chemical properties. The egg shells treated with PA and PAL coatings had a significantly lower microbial population compared to the uncoated egg shells. PA and PAL coatings effectively inhibited the growth of Salmonella after 1 and 7 days of storage, respectively. In addition, no outgrowth was observed up to 42 days.


2018 - Evaluation of the antimicrobial activity of a blend of monoglycerides against Escherichia coli and Enterococci with multiple drug resistance [Articolo su rivista]
Immacolata, Anacarso; Andrea, Quartieri; Riccardo De, Leo; Pulvirenti, Andrea
abstract

Bacterial antibiotic resistance is a natural phenomenon, seriously affecting the treatment of infections. The biggest danger is that current antibiotics are not able to eradicate the resistant strains. In recent years, alternative antibacterial substances are being sought, which can help in these cases. Fatty acids and monoglycerides are known among the natural substances for their antimicrobial properties and, important detail, bacteria do not develop resistance to them. In this work, we studied the antimicrobial effects of a monoglyceride blend against some multi-resistant Enterococci and Escherichia coli strains. Based on literature data, a blend of fatty acids and their monoglycerides was created and its antimicrobial activity was evaluated against 37 strains of E. coli and 17 Enterococci presenting resistance to at least two antibiotics. A different behavior was observed in the two groups of bacteria, proving that alternative substances can be considerate for the potential treatment of multidrug-resistant strains.


2018 - PRESENCE OF ANISAKIS PEGREFFII IN FARMED SEA BASS ( DICENTRARCHUS LABRAX L.) COMMERCIALIZED IN SOUTHERN ITALY: A FIRST REPORT [Articolo su rivista]
Cammilleri, Gaetano; Costa, Antonella; Graci, Stefania; Buscemi, Maria Drussilla; Collura, Rosaria; Vella, Antonio; Pulvirenti, Andrea; Cicero, Antonello; Giangrosso, Giuseppe; Schembri, Pietro; Ferrantelli, Vincenzo
abstract

We examined 151 European sea bass (Dicentrarchus labrax L.) samples from farms and fish markets of Sicily (Southern Italy) for Anisakidae larvae detection. All the samples were examined by visual inspection and modified chloro-peptic digestion. Two nematode larvae were found in the viscera of only one European sea bass sample from a farm located in Greece (FAO 37.3), giving a total prevalence of infestation of 0.7%. No other parasites were found after chloro-peptic digestion of the samples. The larvae were morphologically ascribed, at genus level, to morphotypes I and molecularly identified as Anisakis pegreffii. To the best if our knowledge, this is the first report on the presence of anisakid parasites in farmed European sea bass of Mediterranean Sea. Our findings suggest that the risk of exposure to Anisakidae nematodes in farmed European sea bass remains very low. However, further data on Mediterranean farms are needed to have a detailed risk analysis.


2018 - Properties of poly(vinyl alcohol) films as determined by thermal curing and addition of polyfunctional organic acids [Articolo su rivista]
Bellelli, Massimo; Licciardello, Fabio; Pulvirenti, Andrea; Fava, Patrizia
abstract

The aim of the study was to assess the effect of the addition of citric and malic acid and heat curing on the mechanical, physical and optical properties of poly(vinyl alcohol) (PVOH) films. The addition of the organic acids without successive thermal treatments has a mere plasticising effect, while their application with heat curing has a combined crosslinking and plasticising effect. While conventional plasticizers and crosslinkers improve either extensibility or tensile strength of films, respectively, the addition of citric and malic acid coupled with heat curing determined good tensile strength and extensibility. Hydrophilicity was significantly reduced by thermal curing and even further reduced with the organic acids addition. The high transparency of the PVOH films was not affected either by heat-curing, acid addition and their combination, while the use of high curing temperature coupled with acid addition caused a slight yellowing of the films. The use of citric and malic acid in combination with thermal curing is a viable strategy for tailoring the performances of PVOH films thus broadening their spectrum of application.


2018 - Vegan foods: Mimic meat products in the Italian market [Articolo su rivista]
Bedin, Elisa; Torricelli, Chiara; Gigliano, Silvia; De Leo, Riccardo; Pulvirenti, Andrea
abstract

The current concern about meat consumption tackles many aspects: health, social life, food behaviours, animal welfare, natural resources exploitation. People are more sensitive about these topics and they are shifting individual food habits in favour of a more plant-based diet. Surveys provide a concise view of the increasing percentage of both vegetarians and vegans. The aim of the present research was to develop adequate recipes to prepare food products that mimic the shape and the texture of traditional würstel and Mortadella, focusing on the Italian market. The challenge faced was multiple: firstly, to maintain the similar characteristics of the traditional foods; secondly, to accomplish the consumer's requests; thirdly, to enlarge the market share of the food industries. The results obtained demonstrated that “mimic- würstel” and “mimic-mortadella” were created with vegan allowed ingredients and proteins of vegetal origin. In the case of “mimic-mortadella” the addition of tofu cubes tried to recreate the visual effect of fat globules.


2017 - Detection of food and skin pathogen microbiota by means of an electronic nose based on metal oxide chemiresistors [Articolo su rivista]
Nunez Carmona, Estefania; Sberveglieri, Veronica; Ponzoni, Andrea; Galstyan, Vardan; Zappa, Dario; Pulvirenti, Andrea; Comini, Elisabetta
abstract

This work illustrates the potentiality of a mixed array merging the Nanowire and thin film metal oxide technologies to develop an electronic nose as a tool to monitor the human skin and food pathogen microbiota. Nanowire and thin film sensors have been fabricated, characterized and then integrated together to develop a hybrid device that can enhance the sensing performance. Different blends of microorganisms grown in artificial sweat have been tested. Classical techniques, like GC–MS (Gas Chromatography–Mass Spectrometry) with SPME (Solid Phase Micro Extraction) have been employed in parallel, in order to have a multidisciplinary approach and a consistent data set. The obtained results show the potentiality of the proposed electronic nose to discriminate between the different blends of microorganisms and to follow up microbiota growth inside the blends.


2017 - Elimination of Salmonella cross-contamination on eggs using antimicrobial coating [Poster]
DE LEO, Riccardo; Francesca, Diazzi; Quartieri, Andrea; Pulvirenti, Andrea
abstract

Gastro-intestinal infections caused by Salmonella strains, together with Campylobacter, are among the most common food-borne diseases in Italy (EFSA journal 2016). Therefore, it is important to avoid the spreading of bacterial colonization on different type of foods. A common vector of Salmonella are eggs, which can be contaminated by the laying hen (vertical or horizontal infection) or by contact with contaminated eggs of food (cross-contamination). At the moment, the Italian prevention practices to control Salmonella spreading are bacterial controls on laying hens and their environment and eggs brushing before packaging. Other countries allow eggs washing, but this process decreases the shelf-life of the product. This study investigates the efficiency of an antimicrobial coating to avoid Salmonella cross-contamination on eggs. Coating the eggs with a pectin-alginate polymer drastically improves their food safety and reduces the recovery of Salmonella colonies from the egg surface. Therefore, the occurrence of cross-contamination would be highly reduced as well. The positive effect of the coating itself showed to be highly effective even without the addition of antibacterial agents such as LAE. This active packaging could guarantee higher safety for eggs and opens the possibility to test this procedure on other food matrices common carriers of potentially pathogenic bacteria.


2017 - First report on the presence of Alloxan in bleached flour by LC-MS/MS method [Articolo su rivista]
Vita, Giaccone; Cammilleri, Gaetano; Vita Di Stefano, ; Rosa, Pitonzo; Antonio, Vella; Pulvirenti, Andrea; Gianluigi Maria Lo Dico, ; Vincenzo, Ferrantelli; Andrea, Macaluso
abstract

In this work the presence of Alloxan in bread, pastry and cake bleached flour was investigate in order to verify possible risk for consumers related to the use of chemicals for flour bleaching. A selective UHPLC–MS/MS method has been developed and validated for the purpose. Alloxan is one of the possible minor side products of oxidation after chemical bleaching of wheat flours, when several chemical agents are used. One hundred and seventy-five flour samples were analyzed for Alloxan determination. The validation of the method was performed in accordance with the ISO/IEC/EN 17025 for linearity, detection limit, quantification limit, accuracy, precision and ruggedness determination. Satisfactory performances were obtained for the analyte, with a Limit of Detection (LOD) of 0.73 mg kg−1, a Limit of Quantification (LOQ) of 0.85 mg kg−1 and recovery values between 94% and 102%. The present work report for the first time the presence of trace levels of Alloxan in 24% of the analyzed samples, with mean values of 0,95 ± 0,04 mg kg−1. The presence of Alloxan was detected only in cake flour samples. Further studies on toxicological levels of Alloxan are needed in order to evaluate possible risks for consumers linked to the consumption of bakery products.


2017 - Grated parmigiano reggiano cheese: Authenticity determination and characterization by a novel nanowire device (S3) and GC-MS [Capitolo/Saggio]
Sberveglieri, Veronica; BHANDARI MANOHAR, Prasar; Pulvirenti, Andrea; Carmona, Estefania Núñez
abstract

Parmigiano Reggiano (PR) cheese is produced from the bovine milk in a limited geographic area in northern Italy. It is one of the oldest traditional cheeses manufactured in Europe (XII century) and is still one of the most esteemed Protected Designation of Origin (PDO) cheeses of Italy. The Parmigiano Reggiano Cheese Consortium (CFPR) is the institutional system that covers all the cheese dairy factories and controls the cheese production in terms of cow feeding, cheese manufacture and ripening processes. The aim of this study was to establish a new, rapid, portable, easy-to-use, economic and non-destructive fouling based on nanowire technology device (S3) to control the presence of false grated PR. All the analyzed samples were collected from the Consortium that picked the samples produced in different European countries (three samples from Germany, one from Ireland, one from Latvia and one from Lithuania). The achieved samples were analyzed with the cooperation of multidisciplinary technologies like Colorimeter, pH Meter, GC-MS with SPME and the new nanowire gas sensor device (S3). The applied techniques were able to create a specific database for the S3 device, capable to give results in a few minutes. The use of a Small Sensor System (S3) device to determine the specific volatile organic compounds (VOCs) of the grated PR is a promising approach offering, at the same time, a simpler, faster and easier to handle (no specialized technicians are required) solution with respect to the analytical techniques (GC-MS, HPLC). However, from the commercial point-of-view, low cost and reduced power consumption are the fundamental features that should entail a sensor device. In this study, S3 device was applied and found to be efficient in the discrimination of the different grated PR cheese samples. In particular, the discrimination was checked quali-quantitatively thanks to the analysis performed with the GC-MS-SPME technique. In fact, the achieved results demonstrated the quali-quantitative differences of the VOCs emitted by the samples. S3 device is a promising tool for the quality discrimination of the PR cheese and other foodstuffs. In our future works, specific database of the grated PR will be amplified for the direct use of S3 in the cheese-chain productions.


2017 - LE CERTIFICAZIONI DI QUALITÀ NEL SETTORE AGROALIMENTARE [Altro]
Pulvirenti, Andrea; DE GIORGIO, Roberto
abstract

Le imprese vogliono aumentare la loro credibilità, competitività e reddito. Le richieste del mercato spingono le imprese a investire in competitività attraverso sistemi di certificazione, ispezione e verifica. L'accreditamento è un valore aggiunto per le imprese ed è anche una garanzia di qualità dei processi e dei prodotti e uno strumento fidato per i consumatori. Oggi, i dettaglianti di massa ricorrono sempre più all'accreditamento, che è diventato un'abitudine in tutte le relazioni che richiedono una valutazione di conformità qualificata per proteggere il mercato e i consumatori. Il regolamento (CE) n. 178/2002 ha introdotto il principio di responsabilità di tutti gli operatori della catena alimentare. I rivenditori del mercato di massa (GDO) hanno adottato sistemi di sicurezza e controllo per i loro fornitori. Ciò ha portato allo sviluppo di parametri di riferimento, certificati da organizzazioni di terze parti accreditate, che forniscono requisiti specifici per garantire un elevato livello di sicurezza nei processi di produzione.


2017 - Le certificazioni di qualità nel settore Agroalimentare: stato dell'arte ed evoluzione [RAPPORTO TECNICO] [Altro]
Pulvirenti, Andrea; De Giorgio, Roberto
abstract

Scopo di questo lavoro è di proporre una linea guida e di consulenza per gli operatori del settore alimentare, per la definizione e lo sviluppo di certificazioni e standard ivi comprese la frequenza e tipologia di verifiche svolte per il controllo dello standard stesso. Punto di partenza è stato la valutazione gli obiettivi, i vantaggi, e la lista degli audit al fine di rendere univoci e sostenibili i piani di verifica aziendale, nonché orientare la politica aziendale nel tema della sicurezza alimentare. Secondo la definizione della FAO nel World Food Summit del 1996: “La sicurezza alimentare esiste quando ciascun individuo, in ogni momento, ha accesso a una quantità di cibo sufficiente, sicuro e nutriente in modo da soddisfare i bisogni dietetici e le preferenze alimentari per garantire una vita sana ed attiva” (FAO, 1996). In seguito ad alcuni casi di malattie diffusosi nella popolazione europea negli anni ottanta, imputata alla presenza di ormoni della crescita nella carne bovina, la Comunità europea ha adottato una serie di direttive che vietano tutte le sostanze contenenti ormoni impiegate nell’allevamento di bestiame. In concomitanza con l’approvazione dei primi divieti, in seno al Codex Alimentarius Commission si aprì una discussione sull’opportunità di adottare uno standard sulla presenza di residui di ormoni derivanti da medicinali e antibiotici utilizzati. Mediante standard internazionali come quelli adottati dalla CAC si va a integrare anche il merito delle decisioni i contenuti degli atti di regolazione e di gestione, determinando un impatto su finalità, contenuti e forme dell’amministrazione ivi compresi i diritti e le aspettative degli individui. Il processo di “comunitarizzarzione” della disciplina della sicurezza alimentare risale agli anni settanta del XX secolo. Per salvaguardare la libertà di circolazione dei prodotti alimentari, minacciata dalla difformità delle regole nazionali in materia, la Comunità ha fatto ampio ricorso a misure di armonizzazione normativa e al suo mutuo riconoscimento. Per rendere più trasparente e scientifico il settore della regolamentazione alimentare, alla fine degli anni ’90 è stata avviata una revisione del quadro normativo UE. Nel 1997 è stato messo a punto un nuovo sistema di consulenza scientifica. Oltre al Comitato Direttivo Scientifico, sono state istituite otto nuove commissioni scientifiche. Nel 2002 è stata creata l’EFSA, European Food Safety Authority (Agenzia Europea per la Sicurezza Alimentare), organismo indipendente che lavora in stretta collaborazione con vari enti e istituti scientifici degli stati Membri, offrendo una consulenza scientifica indipendente su tutte le questioni che influiscono direttamente o indirettamente sulla sicurezza alimentare. Inoltre, con la creazione della Food and Agricultural Organization, la materia è divenuta anche di competenza internazionale: “a disciplinare l’agricoltura, sia nella fase della produzione di beni che in quella della loro commercializzazione concorrono oltre al legislatore nazionale, organismi diversi come CE, WTO, ecc. In primo luogo, la disciplina della sicurezza alimentare ha visto la disposizione di carattere regolatori, implicanti poteri di programmazione, autorizzazione, vigilanza, controllo e conformativi, attuati nelle diverse fasi della filiera produttiva. In tal modo la governance del settore non si limita alla tutela del singolo ma è destinata alla generalità dei consumatori di alimenti e finalizzata alla tutela di beni giuridici pubblici come la salute e l’accesso ai cibi sani: l’oggetto della regolazione non riguarda non riguarda solo i rischi che provengono da agenti biologici o fisici ma anche quelli di produzione industriale e chimici; pertanto la sicurezza alimentare non si limita solo a preservare l’igiene e a impedire il deterioramento organico dei cibi, ma è finalizzata a garantire che’ tutti i metodi


2016 - Dessert freddi: criteri per la loro classificazione [Cold dessert: criteria for their classification] [Articolo su rivista]
Chiari, Paola; Pulvirenti, Andrea; Giudici, Paolo
abstract

The expression “cold desserts” names a heterogeneous group of sweet products, which includes gelati, ice cream, pudding, mousse and more. The present work develop an inventory of all the sweet products falling under the definition of “cold desserts”. To reach this aim, several databases (Mintel GNPD), pastry’s books, industrial, as well as artisanal, products are going to be consulted. Moreover, the research has the objective to advance a rational organization of the sweet products comprised by the “cold dessert”’s definition. Thanks to such a logical organization, it will be possible to develop homogenous groupings of products, according to their characteristics. The variables with the greatest discriminating and grouping power employed in this research are: the product’s temperature of consumption, its structure, and the modalities through which it incorporates the air. The main difference between industrial and artisanal gelato is indeed in the modalities through which the air is incorporated in the products and in their consequently different rheological properties.


2016 - Nanowire device (S3) to characterize the genuine aroma of Parmigiano Reggiano cheese [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Carmona, Estefania Nunez; Pulvirenti, Andrea
abstract

Parmigiano Reggiano (PR) is a much-appreciated cheese all over the world although it is produced in a little region in the northeast part of Italy from raw bovine milk.


2016 - Small Sensor Sistem S3 device to control the microbial contamination in water [Relazione in Atti di Convegno]
Carmona, Estefania Nunez; Sberveglieri, Veronica; Ponzoni, Andrea; Zappa, Dario; Pulvirenti, Andrea
abstract

The control and sanitation of water it's a worldwide concerning problem. In particular is in developing countries where the need is more evident due to the lack of sources and appropriate structures to cope with the outbreaks of waterborne diseases. In most of cases, the cause of these significant epidemiological events have microbial origin. Despite of it the situation of water supplies and sanitation has improve all over the world in the last decades. Nanowire technology has already shown their ability to perform very effective and fast monitoring microbiological spoilage and quality control. The aim of this study was test the ability of a novel S3 (Small Sensor System) nanowire device for the detection of complex mixtures of bacteria in potable water in order to approach a real condition in cooperation with GC-MS- SPME technique. The achieved results strongly suggest the application of S3 device as a very easy to use, fast and accurate tool in water quality control.


2015 - Detection of microorganism in water and different food matrix by electronic nose [Capitolo/Saggio]
Sberveglieri, V.; Carmona, E. N.; Pulvirenti, A.
abstract

Nowadays in alimentary industry exist very restrictive quality controls to assure the security of the products and the consumer health. It is assumed that features like the product technology and microbiological quality has been tested when a product arrives on the market. One of the most critical points in food industry today is to avoid microbiological contamination of the products during the different step of the food chain production. The importance of prevention of this episodes relapse not only in the safety of the consumer but also in the prevention of huge economic losses for the industry. On the other hand the presence of microorganisms in food industry is totally required since some kind of them are one of the most important food matrix processors. The current microbiological quality test or those carried out to select the most adequate strains for a determinate target, requires a substantial amount of money, specialized skills and time to reveal the results. The aim of this study was to survey the microbial development trough the Electronic Nose (EN) to be applied in food quality control laboratories. The novel EN used in this study is equipped with an array of chemical gas sensor composed by 6 MOS (Metal oxide sensor) two of this constructed using nanowire technology. This combination of sensors array allow enhancing the threshold of the instrument detecting compounds in low concentration. It also provides information of the presence of a determinate microorganisms and their development in a food matrix reducing drastically the required time for the classical microbiological analysis.


2015 - Nanowire sensor array for the rapid quality control on grated Parmigiano Reggiano [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Carmona, E. Nunez; Pulvirenti, Andrea
abstract

Parmigiano Reggiano (PR) is a much-appreciated cheese all over the world although it is produced in a little region in the northeast part of Italy. The production and elaboration of this kind of cheese is based in a very restrictive normative that is regulated by the “Consorzio del Parmigiano Reggiano”. It is stablish as the 18% as the maximum limit of crust allowed inside the packs of grated PR.


2015 - Nanowire technology to Asses the bacterial presence in water and other food stuff [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Estefania Nunez, Carmona; Pulvirenti, Andrea
abstract

It has been broadly documented the presence of bacteria, yeast and mold in food processing like fermentations, ripening and also spoiling. The early detection of the contamination is critical to preserve the consumer’s health and to avoid economic losses for the industry. The aim of this work was the establish a new fouling based on the cooperation between the use of a novel EN and classical techniques, like GC-MS coupled with classical microbiology, for the detection of bacterial presence in water and other foodstuff.


2015 - Pelleting is a successful method to eliminate the presence of Clostridium spp. from the digestate of biogas plants [Articolo su rivista]
Pulvirenti, Andrea; Ronga, Domenico; Mannella, Lorenzo; Pecchioni, Nicola; Zaghi, Massimo; Tomasselli, Anna Rita; Mannella, Lorenzo; Pecchioni, Nicola
abstract

Biogas production is increasing as a sustainable energy supply, with digestate resulting as a by-product of biogas plants. As a result, the high concentration of Clostridium spp. in digestate became a concern in dairy farming areas. Clostridium spores can contaminate soils and crops when digestate is used as fertilizer, causing a con fl ictual cohabitation of biogas with traditional cheese productions. In order to solve the problem, this study aimed to search for a technical solution enabling either a drastic reduction or the elimination of the content of Clostridium spp. within digestate. Results showed a complete elimination of Clostridium spp. in pelleted stored solid digestate; in addition, pelleting caused a reduction of pH and water mass fraction in terms of fresh weight, and a concentration of mineral nutrients compared to stored solid digestate. Pellet can represent a possible sustainable solution both in reducing potential risks linked to the presence of Clostridium spp. in digestate and in improving the transportation and distri- bution of high-value fertilizer. Hence, pelleting of solid digestate could offer a simple and ef ficient method to allow cohesistence of biogas plants and dairy farming


2015 - Rapid diagnosis of Enterobacteriaceae in vegetable soups by a metal oxide sensor based electronic nose [Articolo su rivista]
Gobbi, Emanuela; Falasconi, Matteo; Zambotti, Giulia; Sberveglieri, Veronica; Pulvirenti, Andrea; Sberveglieri, Giorgio
abstract

A rapid protocol for the early diagnosis of microbial contamination of commercial food products by Electronic Nose is presented. Mixed vegetable soup samples are artificially contaminated by Enterobacter hormaechei and Escherichia coli and a large dataset of 584 samples, over two experimental campaigns, was analyzed by the electronic nose EOS507C based on a four metal oxide sensors array. Diagnosis of the contamination is obtained after 21 h and 18 h from the inoculation of E. hormaechei and E. coli respectively.


2015 - Raw meat shel life increasing by antimicrobial edible coating [Poster]
Pulvirenti, Andrea
abstract

The aim of this work is show how the integration of antimicrobial and edible pectin films from food waste could increase the shelf life of different kind of meat[1]. Chicken and beef raw meat are the two type of meat considered; in two different form: hamburger and sliced meat. To obtain this edible film are used low methoxyl (LM) pectin esterification degree 30-36% and amidation degree 14-20%, in addition with glycerol as plasticizers[2]. The antimicrobial activity are improved with LAE (Ethyl-N-lauryl-L-arginine), 1,6 mg/ml[3]. To observe the real activity of LAE and the migration of that from the coating, all kind of meat are wrapped with different coating: • Pectin film without LAE • Reticulated Pectin film without LAE • Pectin Film with LAE • Reticulated Pectin film with LAE To reticulate the film Calcium D-Pantothenate are added by spraying techniques. All the samples are wrapped in the antimicrobial coating, put inside a Styrofoam tray, closed with polyethylene film, and stored at 4°C. The test consist to check the microbial growth during the storage periods with classic microbiological protocol. 10 grams of sample is mixed with 90 ml of sterile physiological water with a stomacher, the solution formed after serial dilution is inoculated inside Plate count agar media. The bacterial colonies in Petri dish, after 24 hour of incubation at 30°C, are counted to determinate Ufc/g so to establish when meat are not more safe for human health[4]. The final goal of this work is show how the integration of not polluting coating, produced from food waste, had a synergic activity with classic packaging to increase shelf life and safety of high perishable food. The point above means a longer life of the plastic packaging too, then a less production of plastic waste and a better health of environment.


2015 - Skin Microbiota Monitoring by Nanowire MOS Sensors [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Carmona, Estefania Núñez; Ponzoni, Andrea; Comini, Elisabetta; Galstyan, Vardan; Zappa, Dario; Pulvirenti, Andrea
abstract

In this work is illustrated the potentiality use of a Nanowire (Nw) MOS sensor array to be use as tool to monitor the human skin microbiota in order to identify quantitative and qualitative changes. Tree different blends of microorganisms grown in artificial sweat have been tested. Classical techniques, like GC-MS with SPME, have been performed as well, in order to have a multidisciplinary approach and consistent data set. The obtained results show the ability of the NW technology to discriminate between the different blends of microorganisms and to follow up the development of the growth inside the blends during the analysis. The previous results have been confirmed by GC-MS with SPME.


2014 - A Novel Electronic Nose as Adaptable Device to Judge Microbiological Quality and Safety in Foodstuff [Articolo su rivista]
Sberveglieri, Veronica; Nunez Carmona, Estefania; Elisabetta, Comini; Andrea, Ponzoni; Dario, Zappa; Pirrotta, Onofrio; Pulvirenti, Andrea
abstract

This paper presents different applications, in various foodstuffs, by a novel electronic nose (EN) based on a mixed metal oxide sensors array composed of thin films as well as nanowires. The electronic nose used for this work has been done, starting from the commercial model EOS835 produced by SACMI Scarl. The SENSOR Lab (CNR-INO, Brescia) has produced both typologies of sensors, classical MOX and the new technologies with nanowire. The aim of this work was to test and to illustrate the broad spectrum of potential uses of the EN technique in food quality control and microbial contamination diagnosis. The EN technique was coupled with classical microbiological and chemical techniques, like gas chromatography with mass spectroscopy (GC-MS) with SPME technique. Three different scenarios are presented: (a) detection of indigenous mould in green coffee beans, (b) selection of microbiological spoilage of Lactic Acid Bacteria (LAB), and (c) monitoring of potable water. In each case, the novel EN was able to identify the spoiled product by means of the alterations in the pattern of volatile organic compounds (VOCs), reconstructed by principal component analysis (PCA) of the sensor responses. The achieved results strongly encourage the use of EN in industrial laboratories. Finally, recent trends and future directions are illustrated.


2014 - Candida Milleri Detected by Electronic Nose in Tomato Sauce [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Matteo, Falasconi; Emanuela, Gobbi; E., Núñez Carmona; Giulia, Zambotti; Pulvirenti, Andrea
abstract

The tomato sauce is a product of great importance for its massive production in Italy. Microbial contamination is a constant concern for the industries, causing severe economic losses, posing risks to consumers’ health and contributing to an enormous wasting of food. This work shows how the use of the Electronic Nose (EN) EOS 507C can be effective compare to the current procedures in the food production. EN composed of an array of thin film sensors, 6 Metal Oxide (MOX). All the samples were analyzed in parallel with classical chemical technique, like GC-MS with SPME.


2014 - Classification of Different Roasting Processes by MOX Nanowire [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Nunez Carmona, Estefania; Zappa, D.; Comini, E.; Pulvirenti, Andrea
abstract

In the industry of coffee, to obtain a cup of coffee with a balanced aroma, every step in the coffee production chain is crucial one of the most important steps is the roasting process. The roasted coffee is one of the most difficult food matrixes for the complexity of the aroma (VOCs). The aim of this study was to monitor different roasting processes using a novel electronic nose, equipped with an array of MOX gas sensors based on thin films as well as nanowires, in parallel with classical colorimetric techniques to define the homogeneity of the final coffee samples.


2014 - Dalla valutazione qualitativa ai modelli di previsione dei rischi ignoti [Capitolo/Saggio]
Sberveglieri, Veronica; Pulvirenti, Andrea; Giudici, Paolo
abstract

Dalla valutazione qualitativa ai modelli di previsione dei rischi ignoti. L'analisi del rischio come tecnica di valutazione, gestione e comunicazione del rischio era già conosciuta dai primi anni '70, ma solo recentemente ha ricevuto un'attenzione particolare come applicazione specifica nel campo della microbiologia degli alimenti e più in generale, della sicurezza alimentare.


2014 - Detection of microbial contamination in potable water by Nanowire technology [Articolo su rivista]
Carmona, E. N.; Sberveglieri, V.; Ponzoni, A.; Zappa, D.; Pulvirenti, A.
abstract

It is well known that the lack of control and sanitation of water in developing countries has cause very significant epidemiological events. In the last decades the situation of water supplies and sanitation has improve all over the world. Despite of it, in the European Union there are a considerable number of confirmed cases of water-borne infections even though the restrictive law. Electronic Noses (ENs) has shown to be a very effective and fast tool for monitoring microbiological spoilage and quality control. The aim of this study was test the ability of a novel EN for the detection of bacterial presence in potable water in cooperation with analytical (pH) and optical (photometer) techniques. The achieved results notably advocate the use of EN in industry laboratories as a very important tool in water quality control.


2014 - Detection of microbial contamination in potable water by Nanowire technology [Relazione in Atti di Convegno]
Nunez Carmona, Estefania; Sberveglieri, Veronica; A., Ponzoni; D., Zappa; Pulvirenti, Andrea
abstract

It is well known that the lack of control and sanitation of water in developing countries has cause very significant epidemiological events. In the last decades the situation of water supplies and sanitation has improve all over the world. Despite of it, in the European Union there are a considerable number of confirmed cases of water-borne infections even though the restrictive law. Electronic Noses (ENs) has shown to be a very effective and fast tool for monitoring microbiological spoilage and quality control. The aim of this study was test the ability of a novel EN for the detection of bacterial presence in potable water in cooperation with analytical (pH) and optical (photometer) techniques. The achieved results notably advocate the use of EN in industry laboratories as a very important tool in water quality control.


2014 - Electronic nose and gas chromatography-mass spectrometry for the Philippine civet coffee discrimination [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Ongo, Emelda; Falasconi, Matteo; Concina, Isabella; Pulvirenti, Andrea; Sevilla III, Fortunato
abstract

Civet coffee is recognized as the world's most expensive "gourmet" coffee due to its unique taste and aroma. In this work a concrete and promising approach to the headspace profile aroma attributes of Philippine civet coffee using electronic nose (EN) and gas chromatography-mass spectrometry (GC-MS) with SPME techniques was investigated. Chemometric pattern technique was applied to improve the discrimination of civet coffee against its control coffee beans (not eaten by civet animal). EN analysis has shown that aroma characteristic is one of the most important quality indicators of civet coffee. The result was supported by classical chemical techniques like GC-MS analysis with SPME. The chromatographic fingerprints indicated that civet coffees varied with their control beans in terms of composition and concentration of individual volatile constituents (qualitative and quantitative differences). Chemometric discrimination of EN and GC-MS data demonstrated a clearly separated civet from their control coffees, indicating that cultivar and geographic origins decree the aroma and volatile variations in coffee.


2014 - Fast Identification of Different Kind of Green Coffee by Electronic Nose [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Carmona, E. Nunez; Falasconi, M.; Concina, I.; Pulvirenti, Andrea
abstract

The present contribution is focused on the application of the Electronic Nose to the fast detection of fungi in green coffee. The Electronic nose applied in this study is based on array of 6 metal oxide gas sensors with 4 obtained by the film technology and 2 by nanowire technology. The detection of mold in green coffee was achieved by the EN thanks to the cooperation use of different chemical and microbiological (fenotipic) techniques aimed to detect metabolities.


2014 - Fast Identification of Microbiological Contamination in Vegetable Soup by Electronic Nose [Relazione in Atti di Convegno]
Zambotti, G.; Sberveglieri, Veronica; Gobbi, E.; Falasconi, M.; Nunez Carmona, Estefania; Pulvirenti, Andrea
abstract

Microbial contamination, either before or during food production phases, is one of the major concerns of food manufacturers. In this work we present the EOS507C Electronic Nose (EN) for early screening of Enterobacter hormaechei type strain (ATCC 49162) contamination in vegetable soup. The EOS507C, based on an array of metal oxide semiconductor (MOX) sensors, is a rather innovative system equipped with dynamic headspace autosampler and new functionalities such as real-time sample humidity compensation, sensor response linearization and automated periodic calibration. The EOS507C has provided excellent results in terms of screening capabilities: E.hormaechei contamination was detected in 24 hours.


2014 - Microbiological safety in pistachios and pistachio containing products [Articolo su rivista]
Al, Moghazy; M, Boveri; S, ; Pulvirenti, Andrea
abstract

Pistachio nuts, including salted pistachios, unsalted pistachios and shelled pistachios which are ready to eat foods, recently appeared as possible sources of infection with foodborne bacteria besides the high levels of aflatoxins. Several types of pistachio and pistachio containing products were evaluated for the presence of Staphylococcus aureus, and Salmonella spp; also aerobic mesophilic bacteria, coliforms and yeasts and moulds, were enumerated. The analysis was performed in three different sampling times. Aflatoxins levels were determined using a commercial kit.Unsatisfactory levels of mesophilic, coliforms and yeasts and moulds counts were obtained in 55%, 22% and 15% of the samples respectively. Samples contained ≥105CFU/g moulds were tested for aflatoxins. For Salmonella detection, conventional methods were used, and all the suspected colonies on two different selective media were confirmed biochemically. Isolates that biochemically identified as Salmonella spp. were confirmed using multiplex PCR which included primers for 16S rDNA (bacteria control), invA (for Salmonella spp.) and STM4057 (for Salmonella enterica subsp. enterica). Due to presence of 16S rDNA bands only or other bands not explained by any of used primers in multiplex PCR patterns, some isolates were sequenced. Sequencing results revealed that those isolates included Klebsiella pneumonia and Salmonella sp. These results reveled that pistachios and pistachio products can be a possible source of infection with foodborne bacteria and aflatoxins contamination. © 2013 Elsevier Ltd.


2014 - Nanowire Technology for the Detection of Microorganisms in Potable Water [Relazione in Atti di Convegno]
Nunez Carmona, Estefania; Sberveglieri, Veronica; Comini, Elisabetta; Zappa, Dario; Pulvirenti, Andrea
abstract

The lack of surveillance of the water supplies carry out a lot of epidemic issues supplies in developing countries ending in loss of human lives every year, being children the most affected age range. The aim of this study was surveying the microbial quality of water through the Electronic Nose (EN). The novel EN used in this study is equipped with an array of chemical gas sensor composed by 6 MOS (Metal oxide sensor), two of them fabricated using nanowire technology. This sensors array allows enhancing the threshold of the instrument in the detection of compounds in low concentration.


2014 - Poly(vinyl alcohol)-Based Film Potentially Suitable for Antimicrobial Packaging Applications. [Articolo su rivista]
Musetti, Alessandro; Paderni, Katia; Fabbri, Paola; Pulvirenti, Andrea; Al Moghazy, Marwa; Fava, Patrizia
abstract

This work aimed at developing a thin and water-resistant food-grade poly(vinyl alcohol) (PVOH)-based matrix able to swell when in contact with high moisture content food products without rupturing to release antimicrobial agents onto the food surface. This film was prepared by blending PVOH and 7.20% (wt/wt of PVOH) of poly(ethylene glycol) (PEG) with citric acid as crosslinking agent. The film-forming solution was then casted onto a flat surface and the obtained film was 60 μm in thickness and showed a good transparency (close to T = 100%) in the visible region (400 to 700 nm). After immersion in water for 72 h at room temperature, the crosslinked matrix loses only 19.2% of its original weight (the percentage includes the amount of unreacted crosslinking agent, antimicrobial in itself). Water content, degree of swelling, and crosslinking density of the film prove that the presence of PEG diminishes the hydrophilic behavior of the material. Also the mechanical properties of the wet and dry film were assessed. Alongside this, 2.5% (wt/wt of dry film) of grapefruit seed extract (GSE), an antimicrobial agent, was added to the film-forming solution just before casting and the ability of the plastic matrix to release the additive was then evaluated in vitro against 2 GSE-susceptible microorganisms, Salmonella enteritidis and Listeria innocua. The results indicate that the developed matrix may be a promising food-grade material for the incorporation of active substances.


2014 - What happens at the aroma of coffee beans after roasting? [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Pulvirenti, Andrea; E., Comini; Nunez Carmona, Estefania
abstract

The coffee aroma is one of the most important quality evaluation criteria employed for coffee commercialization and consumption. The purpose of this study was following the roasting process VOCs creations with the novel Electronic Nose equipped whit 2 of 6 MOX nanowire sensors. The nanowires exhibit exceptional crystalline quality and a very high length-to-width ratio, resulting in enhanced sensing capability as well as long-term material stability for prolonged operation. Four different methods of roasting, made by ROSTAMATIC (Table 1) machine, were applied to gain a clearer picture of the differences in roasted coffee aromas by means of a volatile compound analysis. Different methods applied on four different origins of green coffee (India, Indonesia, Honduras, Santos and Nicaragua). The commercial coffees products are made from a blending from minimum five different kinds of coffee and the consumers have developed an addiction/expectation to a specific flavor and taste. Different methods of roasting process will provide the coffee different aroma that will add flexibility to those one that already posses the matrix due to different origins. This work tests and illustrates the broad spectrum of potential uses of the EN technique in food quality control.


2014 - What happens at the aroma of coffee beans after roasting? MOX nanowire technology by Novel Electronic Nose to discover the finger print [Articolo su rivista]
Sberveglieri, V.; Pulvirenti, A.; Comini, E.; Carmona, E. N.
abstract

The coffee aroma is one of the most important quality evaluation criteria employed for coffee commercialization and consumption. The purpose of this study was following the roasting process VOCs creations with the novel Electronic Nose equipped whit 2 of 6 MOX nanowire sensors. The nanowires exhibit exceptional crystalline quality and a very high length-to-width ratio, resulting in enhanced sensing capability as well as long-term material stability for prolonged operation. Four different methods of roasting, made by ROSTAMATIC (Table 1) machine, were applied to gain a clearer picture of the differences in roasted coffee aromas by means of a volatile compound analysis. Different methods applied on four different origins of green coffee (India, Indonesia, Honduras, Santos and Nicaragua). The commercial coffees products are made from a blending from minimum five different kinds of coffee and the consumers have developed an addiction/expectation to a specific flavor and taste. Different methods of roasting process will provide the coffee different aroma that will add flexibility to those one that already posses the matrix due to different origins. This work tests and illustrates the broad spectrum of potential uses of the EN technique in food quality control.


2013 - Detection of microorganisms in water and different food matrix by Electronic Nose [Relazione in Atti di Convegno]
Carmona, Estefania Nunez; Sberveglieri, Veronica; Pulvirenti, Andrea
abstract

In the food matrix are involved very elemental products like water to other more complex like, for example, processed and non processed, vegetables and dairy products. All these matrix pass very restrictive controls and a intensive monitoring during the processing time to evaluate their safety and quality. The early detection of the contamination is critical to preserve the consumer's health and to avoid economic losses for the industry. On the other hand, microorganisms are part of a wide range and significant managing process, like fermentations used since ancient times and applied to many different matrix. One of this group of microorganism are Lactic Acid Bacteria (LAB) that play a different role, fermentative or contaminant, depending on the matrix where are founded. Electronic Noses (ENs) has shown to be a very effective and fast tool for monitoring microbiological spoilage and food quality control. The ability of this instrument can also be used for the selection of the most appropriate species or strains for a determinate purpose. The aim of this study was essay the ability of a novel EN for the detection of bacterial presence in water and other foodstuff in cooperation with classical microbiological and chemical techniques like Gas Chromatography Mass Spectrometry with SPME (GC-MS-SPME). The achieved results notably advocate the use of EN in industry laboratories like a very important tool in quality control


2013 - Electronic nose for the early detection of different types of indigenous mold contamination in green coffee [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Comini, Elisabetta; Zappa, Dario; Pulvirenti, Andrea; Carmona, Estefania Nunez
abstract

In the last few years Electronic Noses (ENs) have been revealed to be a very effective and fast tool for monitoring the microbiological spoilage and food quality control. European regulations report the maximum concentration of mycotoxins permitted in green coffee beans. The aim of this study was to test the ability of a novel EN, equipped with an array of MOX gas sensors based on thin films as well as nanowires, to early detect mold contaminations from Aspergillus spp., in cooperation with classical microbiological and chemical techniques like Gas Chromatography coupled with Mass Spectroscopy with SPME technique. In general the selection of the green coffee is controlled by visual inspection of shape, color and size. However, this process in often not enough to prevent the entrance in the food chains of contaminated products. We have demonstrated that the novel EN is able to early detect the qualitative and quantitative differences between contaminate and uncontaminated samples. Achieved results vividly recommend the use of our EN as a quality control tool in coffee producer industry


2012 - Discrimination between different type of coffee according to their country of origin [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Isabella, Concina; Matteo, Falasconi; Pulvirenti, Andrea; Fava, Patrizia
abstract

Geographical origin traceability of food is a relevant issue for both producers’ business protection and customers’ rights safeguard. Between the most widely consumed beverage, coffee is a valuable one, with an aroma constituted by hundreds of volatiles. Differentiation of coffee on the basis of geographical origin still a challenging issue, tough possible by means of chemical techniques. Nonetheless, producer companies need cheaper and simpler tools, able to give a yes/no response in a short time and possibly in a non destructive way. Since the final global volatile composition is also determined by the cultivation climatic conditions, it is in principle possible to distinguish geographical proveniences by exploiting the differences in chemical volatile profile. The present investigation is direct toward the characterization by EN of green and roasted coffees samples according with their geographical origin. The analyzes have been carried out in parallel with chemical classical techniques like GC-MS with SPME. The GC-MS analyses were in good agree with EN results, without sample treatment performed before the analysis.


2012 - Electronic Nose for Microbiological Quality Control of Food Products [Articolo su rivista]
M., Falasconi; I., Concina; E., Gobbi; Sberveglieri, Veronica; Pulvirenti, Andrea; G., Sberveglieri
abstract

Electronic noses (ENs) have recently emerged as valuable candidates in various areas of food quality control and traceability, including microbial contamination diagnosis. In this paper, the EN technology for microbiological screening of food products is reviewed. Four paradigmatic and diverse case studies are presented: (a) Alicyclobacillus spp. spoilage of fruit juices, (b) early detection of microbial contamination in processed tomatoes, (c) screening of fungal and fumonisin contamination of maize grains, and (d) fungal contamination on green coffee beans. Despite many successful results, the high intrinsic variability of food samples together with persisting limits of the sensor technology still impairs ENs trustful applications at the industrial scale. Both advantages and drawbacks of sensor technology in food quality control are discussed. Finally, recent trends and future directions are illustrated.


2012 - Method for the validation of intraspecific crosses of Saccharomycescerevisiae strains by minisatellite analysis [Articolo su rivista]
Boveri, Silvio; Rainieri, Sandra; Pulvirenti, Andrea
abstract

The crossing of Saccharomyces strains by spore conjugation is one of the ways toobtain new starter cultures for the fermentation industry. One of the majordifficulties of this practice is the identification of the newly formed hybrids. In thiswork we describe an effective molecular method for the validation ofSaccharomyces intraspecific crosses. The method described is based in thehypothesis that hybrids constructed by spore conjugation contain the sum of thegenomes of both parental strains. As a consequence, the conjugation of spores oftwo yeasts showing different genomic fingerprinting profiles will result in a hybridculture that will show the sum of both profiles. In this work wWe demonstrated thatthe detection of polymorphism in two genes containing minisatellite-like sequences;,either SED1 or AGA1, is suitable for this purpose. Using this strategy we were ableto validate 15 crosses out of 162 hybridization attempts.


2012 - Monitoring the fermentation process in honey and the evaluationof chemical, microbiological and sensory parameters [Articolo su rivista]
P., Zucchi; Pulvirenti, Andrea; F., De Salvo; G. L., Mazzacan
abstract

In questo lavoro, conclusione di lavori precedentieseguiti su diverse tipologie di mieli monoflora,sono stati riportati i risultati di una sperimentazionecondotta su 7 tipologie di miele. I campioni,messi nelle condizioni ideali per l’innesco dellafermentazione, monitorati per circa un anno, sonostati processati per isolare e identificare i lievitipresenti; valutare temporalmente l’andamento dellivello di etanolo presente nei campioni fermentatie confrontarlo con i risultati ottenuti da un paneldi assaggio incaricato di valutare l’inizio percettivodella fermentazione. L’esito delle ricerche ha messoin evidenza la presenza di Zygosaccharomyces melliscome tipologia di lievito presente e come la sogliapercettiva di etanolo sia da ascriversi intorno ai 100mg/kg per le tipologie di mieli utilizzate in questaricerca.SUMMARYThis work reports the research on 7 monoflora honeys:lime (Tilia spp. L.); clover (Trifolium spp. L.); thyme(Thymus capitatus Hofmgg. et Lk.); thistle (Galactitestormentosa Moench); rhododendron (Rhododendronspp.); tree of heaven (Ailanthus altissima Swingle) treeheather (Erica arborea L.). Honey water was addedat 19%, and the honeys were stored at 30°C untilfermentation. All clover and rhododendron honeyhad fermentation. One of each of the lime honeys,Ailanthus and tree heather did not ferment. Only oneof the thyme and thistle honeys fermented. Ethanoldetermined by enzymatic kit, correlated with sensorialdata. Fermentation was perceived at 100 mg/kg forlight flavoured honeys and at 150 mg/kg for moreflavoured ones. For the microbiological evaluation,4 ground layers were used for yeast checking andidentified by means of PCR/RFLP. The results havehighlighted the presence of only one species of yeastin all of the fermented samples: Zygosaccharomycesmellis. In accordance with the data obtained so far,a content of 50 mg/kg of ethanol is an indicator thathoney is in the fermentation process.


2012 - New methods for the early detection of fungal contamination on green coffee beans by an Electronic Nose [Relazione in Atti di Convegno]
Sberveglieri, Veronica; Fava, Patrizia; Pulvirenti, Andrea; Concina, I.; Falasconi, M.
abstract

Electronic Noses (ENs) are attracting a relevant interest as valuable monitoring tool in several fields, between which the food industry, with special emphasis on microbial contamination detection on food products. Herein we present the ability of an Electronic Nose to early identify the fungi contamination in green coffee. The detection of mold in green coffee was achieved thanks to the cooperative use of different chemical and microbiological (fenotipic) techniques aimed to detect the secondary metabolites


2012 - Ochratoxin A adsorption phenotype: an inheritable yeast trait [Articolo su rivista]
Andrea, Caridi1; Rossana, Sidari; Pulvirenti, Andrea; Giuseppe, Meca; Alberto, Ritieni
abstract

This study aimed to evaluate the inheritance of the trait ochratoxin A adsorption in two wine strains of Saccharomyces cerevisiae and their 46 descendants. Each strain was inoculated intriplicate in test tubes containing 10 mL of must obtained from the Calabrian Zibibbo white grape variety, artificially contaminated with ochratoxin A to reach a total content of 4.10 ng/mL. Themicrovinification trials were performed at 25°C. After 30 days, ochratoxin A values ranged from 0.74 to 3.18 ng/mL, from 0.01 to 2.69 ng/mL, and from 0.60 to 2.95 ng/mL respectively in wines,in lees after washing, and in the saline solution used to wash the lees. The analysis of OTA in wines was performed to find the residual toxin content after yeast activity, thus obtainingtechnological evidence of yeast influence on wine detoxification. The analysis of OTA in lees after washing was performed to distinguish the OTA linked to cells. The analysis of OTA in thesaline solution used to wash the lees was performed to distinguish the OTA adsorbed on yeast cell walls and removed by washing, thus focusing on the adsorption activity of wine yeastthrough electrostatic and ionic interactions between parietal mannoproteins and OTA. Ploidy of the two parental strains was controlled by flow cytometry. Results demonstrated that theochratoxin A adsorption is genetically controlled and is a polygenic inheritable trait of wine yeasts. The majority of the descendants are characterized by a great and significant diversitycompared to their parents. Both the parental strains had genome sizes consistent with their being diploid, so validating the observed results. These findings constitute an initial step to demonstrate the mechanisms of inheritance and establish breeding strategies to improve the ochratoxin A adsorption trait in wine yeasts. This will allow a decrease in the ochratoxin A content of contaminated musts during winemaking, by using genetically improved wine yeasts.


2011 - Application of antimicrobial alginate-oregano essential oil edibile coating on chicken met [Abstract in Atti di Convegno]
Marwa Al, Moghazy; Pulvirenti, Andrea; Fava, Patrizia
abstract

Application of antimicrobial alginate-oregano essential oil edibile coating on chicken met


2011 - DETERMINATION BY ELECTRONIC NOSE OF COFFEE FROM DIFFRENT GEOGRAPHICAL AREAS [Abstract in Atti di Convegno]
Sberveglieri, Veronica; Concina, I.; Falasconi, M.; Gobbi, E.; Pulvirenti, Andrea; Fava, Patrizia
abstract

DETERMINATION BY ELECTRONIC NOSE OF COFFEE FROM DIFFRENT GEOGRAPHICAL AREAS


2011 - DISCRIMINATION BETWEEN DIFFERENT TYPES OFCOFFEE ACCORDING TO THEIR COUNTRY OF ORIGINPresentazione oraleVERONICA SBERVEGLIERI [Abstract in Atti di Convegno]
Sberveglieri, Veronica; Isabella, Concina; Matteo, Falasconi; Pulvirenti, Andrea; Fava, Patrizia
abstract

DISCRIMINATION BETWEEN DIFFERENT TYPES OFCOFFEE ACCORDING TO THEIR COUNTRY OF ORIGIN


2011 - Early detection of fungal contamination on green coffee by a MOX sensors based Electronic Nose [Relazione in Atti di Convegno]
Sberveglieri, Veronica; I., Concina; M., Falasconi; E., Gobbi; Pulvirenti, Andrea; Fava, Patrizia
abstract

ISOEN 2011 - NYEarly detection of fungal contamination on green coffee by a MOX sensors based Electronic Nose


2011 - Identification Of Geographical Origin Of Coffee Before And After Roasting By Electronic Noses [Relazione in Atti di Convegno]
Sberveglieri, Veronica; I., Concina; M., Falasconi; E., Ongo; Pulvirenti, Andrea; Fava, Patrizia
abstract

Identification Of Geographical Origin Of CoffeeBefore And After Roasting By Electronic Noses


2011 - Il processo fermentativo in mieli di sulla ed erba medica:valutazione della specie di lieviti dominante [Articolo su rivista]
Gian Luigi, Mazzacan; Pulvirenti, Andrea; Fausto De, Salvo; Paola, Zucchi
abstract

SUMARYThe aim of our research is toidentify the yeasts responsible forfermentation, the minimum value ofethanol above which fermentationcan be said to begin, and to evaluatethe sensory threshold of fermentationperception related to the value ofethanol and the type of honey. Weanalysed two kinds of honey: Frenchhoneysuckle honey (Hedysarumcoronarium L.); Alfalfa (Medicagosativa L.) from Sardinia.All the alfalfa honeys underwentfermentation but two samples ofFrench honeysuckle honey didnot ferment. The Ethanol contentwas determined by an enzymatickit. For microbiological evaluationfour ground layers were used tocheck for yeasts. The yeasts wereidentified by PCR/RFLP of the ITSregions. Zygosaccharomyces melliswas present in all. The Ethanolcontent was correlated with sensorialanalysis data. The expert assessorsdetect fermentation at a low level ofethanol. Fermentation was alreadyperceived at just 90 mg/kg of ethanol.It can be stated that a content of 50mg/kg of ethanol shows the start ofthe fermentation process.SOMARIOSi sono identificati i lievitiresponsabili della fermentazione,il livello di etanolo, e la soglia diriconoscimento sensoriale. Utilizzati2 mieli italiani: sulla (Hedysarumcoronarium L.); erba medica (Medicagosativa L.). I campioni di medicasono fermentati, mentre 2 campionidi sulla non sono fermentati. Imieli furono analizzati con Kitenzimatico per valutare l’etanoloprodotto: bisettimanalmente finoal raggiungimento di 50 mg/kgdi etanolo, poi settimanalmente.Sono stati usati 4 tipi di terrenoper l’isolamento dei lieviti osmofiliquindi le analisi molecolari perestrarre il DNA. I lieviti isolati sonostati identificati mediante l’analisiPCR/RFLP delle regioni ITS conenzimi di restrizione HaeIII eHinf I. Il lievito presente è statoZygosaccharomyces mellis. L’etanoloè stato correlato con i dati ottenutidall’analisi sensoriale: gli assaggiatorihanno rilevato la fermentazione giàa 90 mg/kg: si può affermare chevalori di etanolo prossimi ai 40-50mg/kg esprimono l’innesco dellafermentazione.


2011 - Microbiological safety and moleculaar analysis of Klebsiella ssp. and Salmonella ssp. Isolated from different pistachio's product [Abstract in Atti di Convegno]
Marwa Al, Moghazy; Boveri, Silvio; Pulvirenti, Andrea
abstract

Microbiological safety and moleculaar analysis of Klebsiella ssp. nd Salmonella ssp. Isolated from different pistachio's product


2011 - SETTIN UP A NEW METHOD FOR RAPID IDENTIFICATION OF FUNGAL CONTAMINATION ON GREEN COFFEE BY MOX SENSORS BASED ELETRONIC NOSE [Abstract in Atti di Convegno]
Sberveglieri, Veronica; Concina, I.; Falasconi, M.; Gobbi, E.; Pulvirenti, Andrea; Fava, Patrizia
abstract

SETTIN UP A NEW METHOD FOR RAPID IDENTIFICATION OF FUNGAL CONTAMINATION ON GREEN COFFEE BY MOX SENSORS BASED ELETRONIC NOSE


2010 - Shelf-life improvement of horse meat by preservative lactic cultures [Abstract in Atti di Convegno]
Pulvirenti, Andrea; Boveri, Silvio
abstract

The aim of this work was to improve the Shelf-life of horse meat by applying lactic acid bacteria starter cultures directly on the meat samples. We selected some lactic cultures by isolating strains directly from pork sausage, and by identifying good phenotypic and metabolic traits, such as, lactic antagonism, good acidification and non-producing bacteriocins. These traits were exploited to reduce the indigenous, spoilage and pathogens microorganisms present in horse meat thus improving the Shelf-life.In particular, a Lactobacillus sakeii classified as GRAS (generally recognized as safe) was selected; the isolate was identified by phenotypic and molecular methods.The action of L. sakeii was assessed over 21 days the correspond to the entire commercial time of horse meat required by Italian large scale retail distribution. Microbiological analysis (aerobic mesophilic and psycrophilic count, lactic bacteria count and the search of clostridia) and sensorial analysis were carried out on two kind of meat cuts: minced and sliced horse meat inoculated with L. sakeii suspension and packaged in MAP condition (60% CO2, 40% N2). The results obtained showed that minced and slices horse meat treated with the lactic starter cultures preserved the sensorial and microbiological characteristics for over 21 days when stored at 4°C.Based on commercial and microbiological parameters, the inoculated horse meat showed good freshness property.


2009 - Aspetti qualitativi del miele fermentato [Articolo su rivista]
Pulvirenti, Andrea; P., Zucchi; F., Desalvo; G., Marcazzan; A. G., Sabatini; Giudici, Paolo
abstract

La fermentazione alcolica ad operadei lieviti è l’unica alterazionemicrobiologica che può subire ilmiele, con conseguenze negativeirreversibili sulla qualità delprodotto.L’elevato contenuto d’acqua delmiele è una condizione necessariaallo sviluppo dei lieviti. Mieli checontengono meno del 17-18% diacqua sono poco suscettibili allafermentazione, la cui probabilitàaumenta con l’aumentare delcontenuto d’acqua. In questo lavoro21 campioni di miele di diversaprovenienza sono stati conservatie regolarmente monitorati per unperiodo di 12 mesi, alla fine delquale sono stati isolati ed identificatii lieviti presenti. Nei mieli sonostate isolate tre specie di lievito:Rhodotorula glutinis, Candidamagnoliae, Hanseniaspora uvarum.Quest’ultima specie è stata isolata neicampioni che presentavano evidentisegni di fermentazione (presenzadi alcool). Inoltre, la presenza dietanolo è stata correlata con i risultatidell’analisi sensoriale.


2009 - Colture lattiche protettive e miglioramento della shelf-life della carne equina [Articolo su rivista]
Pulvirenti, Andrea; Boveri, Silvio; M., Masina
abstract

Colture lattiche protettive e miglioramento della shelf-life della carne equina


2009 - Microbiologia degli Alimenti [Traduzione di Libro]
Pulvirenti, Andrea
abstract

Questo volume rappresenta la versione italiana della (TM)ultima edizione di uno dei testi piA autorevoli e completi sulla microbiologia degli alimenti a " Modern Food Microbiology a " giA tradotto in varie lingue, tra le quali cinese e hindi. La trattazione introduce i fattori intrinseci ed estrinseci che influenzano la crescita microbica negli alimenti e quindi approfondisce il ruolo e la rilevanza dei diversi microrganismi prendendo in esame le principali categorie di prodotti alimentari, compresi quelli di quarta gamma e pronti al consumo. Una parte del volume A] specificamente dedicata alle tecniche di ricerca dei microrganismi e dei loro metaboliti, dalle metodiche tradizionali a quelle piA avanzate. I diversi aspetti e le problematiche della conservazione degli alimenti sono trattati in relazione alle tecniche disponibili e ai fattori e alle forme di resistenza dei diversi gruppi microbici. Sono inoltre approfonditi i temi della valutazione e della (TM)analisi del rischio e degli indicatori di qualitA e di sicurezza in tutte le fasi della produzione alimentare. Conclude il volume una (TM)esaustiva rassegna delle principali malattie trasmesse da alimenti, dei patogeni responsabili e delle misure di controllo e prevenzione. La (TM)opera A] ricca di illustrazioni, tabelle e grafici e ogni capitolo A] completato da una (TM)ampia bibliografia. Un testo indispensabile per gli studenti e i ricercatori, ma anche un prezioso strumento di lavoro e di consultazione per tutti coloro che operano professionalmente nel settore alimentare o a stretto contatto con esso.


2009 - Optimizing the selection process of yeast starter cultures by preselecting strains dominating spontaneous fermentations [Articolo su rivista]
Pulvirenti, Andrea; Rainieri, Sandra; Boveri, Silvio; Giudici, Paolo
abstract

We propose an efficient and time-saving strategy for starter culture selection. Our approach is based on the accomplishment of 3 phases: (i) the selection of yeast strains dominating spontaneous fermentations, (ii) the selection among the dominant strains of those showing the best technological characteristics, and (iii) the final selection among good technological strains of those showing the desired qualitative traits. We applied this approach to wine fermentations, even though the same strategy has the potential to be employed for the selection of any type of starter culture. We isolated and identified yeast strains at the mid- and final stages of 6 spontaneous fermentations carried out in 3 different Spanish wineries. We identified all strains as Saccharomyces cerevisiae by restriction fragment length polymorphism of the ribosomal DNA internal transcribed spacer region, and subsequently distinguished each strain by analyzing the polymorphism of the inter-δ regions. Strains that were detected both at the mid- and final stages of the fermentation were considered dominant. Four dominant strains were finally selected and tested in pilot-scale fermentation, and their performance was compared with that of a commercial wine strain. All dominant strains showed good fitness and resulted suitable to be employed as starter cultures. One of the dominant strains isolated in this study is currently commercialized.


2009 - Selection of Saccharomyces cerevisiae strains with high ability to adsorb ochratoxin A during fermentation [Abstract in Atti di Convegno]
Giuseppe, Blaiotta; Valentina, Coppola; Alberto, Ritieni; Pulvirenti, Andrea; Andrea, Caridi
abstract

Ochratoxin A (OTA) exerts several toxic effects, mainly involving kidney and liver. Itsoccurrence in wines, mainly in the red ones, has been widely reported (Zimmerli andDick, 1996). Recent studies prove the possibility to decontaminate grape musts withselected yeasts able to remove OTA during winemaking (Bejaoui et al., 2004; Caridiet al., 2006); this ability seems related to the adsorption on the outermost layer of theyeast celi wall and has been associated to the parietal mannoproteins, whosecontent in S. cerevisiae depends on the specific yeast strain (Caridi, 2006). In thisstudy, 120 strains of S. cerevisiae coming from three Italian collections [Portici(Naples), Reggio Calabria and Reggio Emilia] previously characterized for theirphysiological, technologieal and genetic features were tested in vitro for OTA removalcapacity in synthetic rnust. Pre-cultures were prepared in YPD at 28'C for 48 h. Testswere performed in triplicate on 10 mi of synthetie must artificially contaminated wilh 5ppb of OT A. After inoculums with 0.2 mi of YPD pre-culture, tubes were incubated at25°C for 21 days. Residual OTA in wines was determined by HPLC as alreadydeseribed by Caridi et al. (2006), analyzing three aliquots of each tesi tube. In aglobal view of results oblained it seams that S. cerevisiae strains are able to removeabout 45% of OTA during fermentalion (the mean value of the 120 wines was45.75% ± 6.29). By contrast, analyzing in detail the results it is evident a highvariability: some strains were able to remove less Ihan 30% of OTA, others reachedremoval levels of 55-60%. The occurrence of strains showing an OTA removal majorthan 55%, and with a low standard deviation, were very low: a} two strains out 38 ofthe Portici colleclion (% of OTA removal: 58.83 ± 1.12 and 57.13 ± 4.42,respectively); b) one strain out 18 of the Reggio Emilia colleclion (% of OTA removal:61.08 ± 1.09); c) three strain out 64 of the Reggio Calabria colleetion (% of OTAremoval: 57.96 ± 2.68, 57.35 ± 1.99, and 57.19 ± 3.88, respectively). ReferencesCaridi et al. (2006). Enzyme and Microbial Technology 40, 122-126. Caridi (2006).Antonie van Leeuwenhoek 89, 417-422. Bejaoui et al. (2004). Journal of AppliedMicrobiology 97, 1038-1044. Zimmerli and Dick (1996). Food Additives andContaminants 13, 655-68.


2009 - Wine strain improvement strategies to enhance red wine safety based on parietal adsorption activity [Abstract in Atti di Convegno]
Andrea, Caridi; Rossana, Sidari; Pulvirenti, Andrea; Giuseppe Blaiotta e., Alberto Ritieni
abstract

There is a great diversity among yeasts for parietal adsorption activity: the outermostlayer of the celi wall notably varies for composition and structure. Parietalmannoproteins can contain negative charges that modify electrostatic and ionicinteraction with wine components, giving good reason to propose a specific selectionof wine yeasts, based on type and amount of parietal mannoproteins. Phenoliccompounds interact with yeast mannoproteins; they can diminish the total polyphenolindex of wine by interaction between anthocyanins and celi walls. Wine yeastinfluences concentration and composition of phenolic compounds in wine, above aliby adsorbing them on celi wall. Remarkable correlations between yeast used forwinemaking and phenolic compounds in wine were reported, demonstrating thatstrain behaviour can modify chromatic properties, phenolic profile and antioxidantpower of wine. The parietal adsorption of grape pigments during winemaking hasimportant effects for wine quality, because of the amount of pigment removed. Yeastcan also interact with wine colour producing anthocyanin-beta-D-glucosidase, pyruvicacid, acetaldehyde, and polysaccharides. Wine yeasts can also selectively removeochratoxin A (OTA) during winemaking because they possess the ability to adsorbOTA. Based on parietal adsorption activity, a specific selection of yeasts will beperformed to protect wine colour and phenolic compounds responsible for antioxidantactivity of wines but to remove OTA. Genomic strategies for genetic improvement ofstrains will be used to obtain a further enhancement of the adsorption/non-adsorptionactivity of wine yeasts. The production of new strains through sexual recombinationcan be achieved through two different approaches: gametes derived from cropsindividuai spores of parental strains gametes or directly from the parental strains. Theinheritable nature of the adsorption of wine colour was recently analysed on thedescendants derived fram three wine strains of S. cerevisiae. Investigation of theprageny demonstrated that the adsorption of wine colour is a polygenic inheritablequantitative traits loci, partially and interdependently correlated to colour and phenoliccontent of wines. This work was supported by PRIN 2007 "Wine strain improvementstrategies to enhance red wine safety based on parietal adsorption activity".


2009 - Wine strain improvement strategies to enhance red wine safety based on parietal adsorption activity. Selection of Spanish, Portuguese and Sicilian wine yeasts. [Abstract in Atti di Convegno]
Caterina Vadalà, Andrea Carldi Rossana S. i. c. a. r. i.; Pulvirenti, Andrea
abstract

Aim of this study was lo perform a specific selection of yeasts for red winemaking.Using a phenotypic-based methodology 99 slrains of Saccharomyces cerevisiae ablelo easily sporify, low H2S-producing, low acetic acid-producing, noi toam producingand exhibiting high fermenlalion power during red winemaking have been selecledfrom a major yeast collection. In a second slep, a Petri plate evaluation of wine colouradsorplion phenolype of yeasls and the result confirmation by microvinificalion trialswere carried out. Strains were split based on Iheir ability to adsorb phenolic grapepigments lo identify a maximum of ten strains, the lowest adsorbing ones. The 99strains of S. cerevisiae were isolated from Spanish, Portuguese and Sicilian mustsand wines and are included in the Department of "Scienze Agrarie e degli Alimenti"collection. The yeasts were studied in Petri plates for the following biochemicalcharacteristics: spore production on acetate agar at 25°C for 10d, H2S production onBiGGY agar at 25°C for 48h, aceti c acid production on calcium carbonate agar at25°C far 7d, adsorption of grape pigments on grape skin agar al 28°C far 10d (Caridiet al., 2002; 2007). The microvinification Irials were performed using Calabrian blackgrapes from Ihe cultivar Gaglioppo; Ihe grapes were destemmed, crushed andsoaked at O°C for 3 days, performing a punch down Iwice per day. The mustobtained after pressing was divided in aliquols of 20 mi and fermenled at 20°C withthe 99 pre-selecled yeasts inoculaled at 5% in triplicate. Analyses were performed ondiluled wine and values were reported considering Ihe dilution. Significant differencesfor each parameter were observed. Biomass colour varied from 51 to 129 (redcomponent), from 27 lo 88 (green component), and from 31 to 82 (blue component).Wine parameters vary from 0.782 to 3.480 (420 nm), from 0.497 to 2.105 (520 nm),from 0.148 to 0.950 (620 nm), from 1.452 to 6.535 (intensity), from 1.304 lo 2.040(tint), from 6.150 to 25.483 (Folin-Ciocalteu index). Based on these results, parietaladsorption activity is notably different among yeasts and justifies a programme ofgenelic improvemenl. References Caridi A. et ai., (2002), J Gen Appl Microbiol, 48,261 Caridi A et al., (2007), J Appl Microbiol, 103,735. This work was supported byPRIN 2007 "Wine slrain improvement strategies lo enhance red wine safety based onparietal adsorption activity".77


2009 - Zygosaccharomyces mellis specie dominante nei processi fermentativi in mieli di agrumi, castagno, girasole, melata. [Articolo su rivista]
Pulvirenti, Andrea; Boveri, Silvio; P., Zucchi; F., Desalvo; G., Marcazzan
abstract

Zygosaccharomyces mellis specie dominante nei processi fermentativi in mieli di agrumi, castagno, girasole, melata.


2008 - Diversity of lactic acid bacteria population in ripened Parmigiano Reggiano cheese [Articolo su rivista]
Gala, Elisabetta; Landi, Sara; Solieri, Lisa; Nocetti, M; Pulvirenti, Andrea; Giudici, Paolo
abstract

The diversity of dominant lactic acid bacteria population in 12 months ripened Parmigiano Reggiano cheeses was investigated by a polyphasic approach including culture-dependent and independent methods. Traditional plating, isolation of LAB and identification by 16S rDNA analysis showed that strains belonging to Lactobacillus casei group were the most frequently isolated. Lactobacillus helveticus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus parabuchneri, and Lactobacillus buchneri species were detected with lower frequency. PCR-denaturing gradient gel electrophoresis (DGGE) applied to DNA extracted directly from cheese samples and sequencing of rDNA amplicons confirmed the complex microbiological pattern of LAB in ripened Parmigiano Reggiano cheeses, with the significant exception of the Lactobacillus fermentum species, which dominated in several samples, but was not detected by cultivation. The present combination of different approaches can effectively describe the lactic acid bacteria population of Parmigiano Reggiano cheese in advanced stages of ripening, giving useful information for elucidating the role of LAB in determining the final cheese quality.


2008 - LA PREVALENZA DEI LIEVITI STARTER IN ENOLOGIA [Articolo su rivista]
Pulvirenti, Andrea; M., De Paola
abstract

In una fermentazione alcolicasi ha prevalenza delceppo di lievito inoculatosui lieviti indigeni quando ilprodotto ottenuto mostra le caratteristichedi un mosto fermentatoin purezza, cioè senzaaltri lieviti presenti, e i risultatiottenuti sono quindi coerenticon le aspettative. Diversi lavoriscientifici hanno messo in luceche è possibile dimostrare laprevalenza del ceppo inoculato,data dal rapporto numericofra le cellule del ceppo inoculatoe quelle dei lieviti indigenidel mosto


2008 - Ochratoxin A adsorption phenotype: an inheritable yeast trait [Abstract in Atti di Convegno]
Andrea, Caridi; Rossana, Sidari; Pulvirenti, Andrea; Giuseppe, Mecca; Alberto, Ritieni
abstract

21st International ICFMH Symposium “Evolving Microbial Food Quality and Safety” Aberdeen 1st – 4th September


2008 - Studio dell’evoluzione della popolazione di batteri lattici durante la produzione di Parmigiano Reggiano mediante 16S rDNA PCR-DGGE [Articolo su rivista]
DE VERO, Luciana; Solieri, Lisa; Gala, Elisabetta; Pulvirenti, Andrea; Giudici, Paolo
abstract

Il formaggio Parmigiano Reggiano è stato oggetto di numerosi studi microbiologici, in particolare sulle prime fasi di produzione e sul siero innesto. Nella presente ricerca, è stata studiata l’evoluzione delle specie di batteri lattici (BL) durante tutto il processo compresa la stagionatura. A tal fine è stata utilizzata una tecnica coltura indipendente, che prevede l’estrazione del DNA batterico direttamente dalla matrice, senza pertanto l’isolamento e la coltura dei batteri. Successivamente, parte della regione 16S del DNA ribosomale (rDNA) è stata amplificata con primers specifici per alcune specie di BL e gli amplificati ottenuti sono stati separati mediante gel elettroforetico di poliacrilammide in gradiente denaturante (PCR-DGGE). In letteratura è riportato che, sia nella produzione di Parmigiano Reggiano che di Grana Padano, Lactobacillus helveticus è la specie microbica prevalente nel siero innesto, fin oltre il 70% della popolazione microbica. Al contrario, nella nostra ricerca, abbiamo osservato che Lb. helveticus è prevalente nel siero maturo ma non nel siero a fine lavorazione. In particolare, durante il processo in caldaia si ha un recupero di Lactobacillus delbrueckii che numericamente prevale sulla specie Lb. helveticus. Successivamente, il siero raccolto e conservato per la preparazione della coltura starter della lavorazione successiva, favorisce la crescita di Lb. helveticus, ripristinando la dominanza di quest’ultima specie. In generale, la tecnica della PCR-DGGE può essere ritenuta maggiormente attendibile nella descrizione delle specie maggioritarie rispetto ai metodi coltura dipendente, in quanto consente di superare i problemi relativi alla non coltivabilità delle cellule batteriche presenti nel mezzo pur essendo esse in uno stato vitale.


2008 - The inheritance of mtDNA in lager brewing strains [Articolo su rivista]
Rainieri, Sandra; Kodama, Yukiko; Nakao, Yoshihiro; Pulvirenti, Andrea; Giudici, Paolo
abstract

n this work, we compared the mtDNA of a number of interspecific Saccharomyces hybrids (Saccharomyces cerevisiae x Saccharomyces uvarum and S. cerevisiae x Saccharomyces bayanus) to the mtDNA of 22 lager brewing strains that are thought to be the result of a natural hybridization between S. cerevisiae and another Saccharomyces yeast, possibly belonging to the species S. bayanus. We detected that in hybrids constructed in vitro, the mtDNA could be inherited from either parental strain. Conversely, in the lager strains tested, the mtDNA was never of the S. cerevisiae type. Moreover, the nucleotide sequence of lager brewing strains COXII gene was identical to S. bayanus strain NBRC 1948 COXII gene. MtDNA restriction analysis carried out with three enzymes confirmed this finding. However, restriction analysis with a fourth enzyme (AvaI) provided restriction patterns for lager strains that differed from those of S. bayanus strain NBRC 1948. Our results raise the hypothesis that the human-driven selection carried out on existing lager yeasts has favored only those bearing optimal fermentation characteristics at low temperatures, which harbor the mtDNA of S. bayanus


2006 - I microrganismi dell'aceto balsamico. [Esposizione]
Giudici, Paolo; Solieri, Lisa; Gullo, Maria; Pulvirenti, Andrea; DE VERO, Luciana; Falcone, Pasquale Massimiliano
abstract

Il processo di produzione dell’aceto balsamico tradizionale prevede quattro fasi distinte: cottura del mosto, fermentazione alcolica, fermentazione acetica ed invecchiamento. Ognuna di queste fasi va gestita attentamente sulla base di fattori sia di tipo chimico-fisico che microbiologico. Data la scalarità delle fasi, le anomalie presenti a monte del processo si ripercuotono sul prodotto finale. Ad esempio, le fermentazioni spontanee sono tra le principali cause di variabilità e di insuccesso ed è per tal motivo che occorre un accurato monitoraggio della batteria che possa preventivamente correggere eventuali irregolarità nel sistema. In particolare, i parametri da monitorare durante la gestione della batteria sono: l’acidità titolabile, la concentrazione alcolica, il pH, la temperatura e l’ossigeno. L’impiego di colture starter di lieviti e batteri acetici selezionate sulla base dei caratteri desiderabili può essere utile per la standardizzazione del processo e per l’ottenimento di un prodotto che presenti le proprietà chimico-organolettiche richieste. L’età dell’aceto balsamico tradizionale è difficile da definire e da calcolare per il particolare modo di produzione: aliquote di aceto sono trasferite annualmente da un barile all’altro. La conduzione delle batterie rende difficoltoso il calcolo dell’età, tuttavia in questo lavoro abbiamo sviluppato un modello algebrico per il calcolo dell’età. In particolare abbiamo definito i limiti di età teoricamente raggiungibili in ogni barile della batteria.


2006 - Le Fermentazioni dell’Aceto Balsamico Tradizionale [Capitolo/Saggio]
Giudici, Paolo; Gullo, Maria; Solieri, Lisa; DE VERO, Luciana; Sara, Landi; Pulvirenti, Andrea; Sandra, Rainieri
abstract

Scritto a più mani dall'équipe di ricerca dell'Università di Modena e Reggio Emilia che fa capo al professor Paolo Giudici, il libro presenta al pubblico degli amanti dell'aceto balsamico tradizionale i più recenti risultati delle ricerche svolte in ambito microbiologico nelle varie fasi di produzione di questo straordinario prodotto della nostra tradizione, amato e apprezzato in tutto il mondo. Il professor Giudici in questo volumetto articola con limpida chiarezza divulgativa parte dei risultati conseguiti dalla sua équipe, trasmettendo alcune nozioni necessarie per la conduzione delle fermentazioni. A ciò si aggiunga una sezione di letteratura specialistica particolarmente abbondante e curata sulla microbiologia dell'aceto balsamico.


2006 - Le fermentazioni dell’aceto balsamico tradizionale [Monografia/Trattato scientifico]
Giudici, Paolo; Gullo, Maria; Solieri, Lisa; DE VERO, Luciana; Landi, Sara; Pulvirenti, Andrea; Rainieri, Sandra
abstract

Scritto a più mani dall'équipe di ricerca dell'Università di Modena e Reggio Emilia che fa capo al professor Paolo Giudici, il libro presenta al pubblico degli amanti dell'aceto balsamico tradizionale i più recenti risultati delle ricerche svolte in ambito microbiologico nelle varie fasi di produzione di questo straordinario prodotto della nostra tradizione, amato e apprezzato in tutto il mondo. Il professor Giudici in questo volumetto articola con limpida chiarezza divulgativa parte dei risultati conseguiti dalla sua équipe, trasmettendo alcune nozioni necessarie per la conduzione delle fermentazioni. A ciò si aggiunga una sezione di letteratura specialistica particolarmente abbondante e curata sulla microbiologia dell'aceto balsamico.


2006 - Mitochondrial DNA and Mitochondrial proteins in interspecific Saccharomyces hybrids [Relazione in Atti di Convegno]
Pulvirenti, Andrea; Solieri, Lisa; DE VERO, Luciana; Giudici, Paolo
abstract

Mitochondrial DNA and Mitochondrial proteins in interspecific Saccharomyces hybrids


2006 - Strategie di selezione e miglioramento genetico [Articolo su rivista]
Giudici, Paolo; Rainieri, Sandra; Pulvirenti, Andrea; NUOVI APPROCCI, CARLO Z. A. M. B. O. N. E. L. L. I. . TRADIZIONE E.
abstract

This work report the basis of wine yeastselection. Moreover it shows a rationaland critic comparison of the importanceof selected starter yeast cultures vsautochthones yeast culture, whose scientificvalue has recently been overestimated.Finally it describes the major geneticimprovement techniques and propose someinnovative improvement approaches thatinvolve the entire yeast genome. Theseapproaches have the potential to efficientlyreach the improvement goals set for theoenological sector, which in most casesrefer to characters that are regulated by ahigh number of genes randomly distributedthrough the entire genome.In questo lavoro sono illustrate le basi su cuisi fonda il processo di selezione per i lieviti diinteresse enologico. Viene inoltre presentatauna visone razionale e critica sull’importanzadelle colture di lievito selezionate rispettoalle colture autoctone, il cui valorescientifico ultimamente sembra essere statosopravalutato. Infine vengono descritte leprincipali tecniche di miglioramento geneticotradizionali e proposti approcci innovativi checoinvolgono l’intero genoma. Questi approccihanno il potenziale di raggiungere in modoefficace gli obiettivi di miglioramento tipicidel settore enologico, che nella gran partedei casi riguardano caratteri regolati da unnumero molto elevato di geni distribuiti inmodo casuale nell’intero genoma.


2006 - The Yeasts of sourdough [Relazione in Atti di Convegno]
Pulvirenti, Andrea; Solieri, Lisa; Sara, Landi; Giudici, Paolo
abstract

III International Symposium on Sourdough. The yeasts of Sourdough


2005 - Limitations on the use of polymerase chain reaction restriction fragment length polymorphism analysis of the rDNA NTS2 region for the taxonomic classification of the species Saccharomyces cerevisiae. [Articolo su rivista]
Pulvirenti, Andrea; Solieri, Lisa; DE VERO, Luciana; Giudici, Paolo
abstract

Different molecular techniques were tested to determine which was the most effective in the identification of Saccharomyces cerevisiae strains. In particular, polymerase chain reaction--restriction fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer (ITS) regions and the nontranscribed spacer 2 (NTS2) region, sequencing of the D1/D2 domain, and electrophoretic karyotyping were applied to 123 yeast strains isolated from different sourdoughs and tentatively attributed to the species S. cerevisiae. All of the strains tested showed an identical PCR-RFLP pattern for the ITS regions, an identical nucleotide sequence of the D1/D2 domain, and the typical electrophoretic karyo type of S. cerevisiae. In contrast, 14 out of the 123 strains tested showed some polymorphism with BanI restriction analysis of the NTS2 region. Our results indicate that while the sequencing of the D1/D2 domain, the PCR-RFLP analysis of the ITS regions, and the electrophoretic karyotype can be employed successfully to identify S. cerevisiae strains, PCR-RFLP analysis of the NTS2 region does not allow a consistent and accurate grouping for S. cerevisiae strains. The fact that the NTS2 region of a small number of strains (8.78% of the total strains tested) is different from that of the other S. cerevisiae strains confirms that molecular methods should always be tested on a great number of strains.


2005 - Molecular procedure for yeasts identification of Traditional Balsamic Vinegar [Relazione in Atti di Convegno]
Solieri, Lisa; L., Castellari; A. R., Battagliola; Pulvirenti, Andrea; S., Landi; Giudici, Paolo
abstract

Molecular procedure for yeasts identification of Traditional Balsamic Vinegar


2005 - Screening for yeasts able to hydrolyse arbutin in the presence of glucose or ethanol [Articolo su rivista]
A., Caridi; Pulvirenti, Andrea; C., Restuccia; R., Sidari
abstract

The study was carried out using 212 glucose-fermenting yeasts isolated from Calabrian and Sicilian samples of must and wine. They were screened for beta-glucosidase activity in Petri plates containing arbutin agar medium. Eleven strains (three apiculate yeasts and eight elliptic yeasts) were able to perform arbutin hydrolysis, but only strain C5, identified as Pichia anomala, maintained the ability to hydrolyse arbutin even in the presence of remarkable amounts of glucose or ethanol. This strain did not show the capacity to excrete the beta-glucosidase into the medium; effectively, the enzymatic activity in the culture supernatant was near to zero. However, its intracellular activity was high.


2005 - Strategies and perspectives for genetic improvement of wine yeasts. [Articolo su rivista]
Giudici, Paolo; Solieri, Lisa; Pulvirenti, Andrea; Cassanelli, Stefano
abstract

Recent developments in expression profile and proteomic techniques illustrated that the main oenological traits of wine yeasts are complex and influenced by several genes, each of them identified as absolutely essential. Only for monogenic properties the genetic improvement programmes of wine yeasts can be performed by alteration of individual genes. Ideally the most productive way of improving the whole-cell biocatalysts is by evolution of the entire cell genome. In this article we briefly review the main genetic improvement techniques applied in new and optimised wine strains construction, paying particular attention to blind and whole genome strategies, such as the sexual recombination and genome shuffling.


2005 - The yeasts of Traditional Balsamic Vinegar [Abstract in Atti di Convegno]
Solieri, Lisa; L., Castellari; A. R., Battagliola; Pulvirenti, Andrea; S., Landi; Giudici, Paolo
abstract

Proocedng of Congress: International Symposium on Vinegars and Acetic Acid Bacteria


2005 - vinegars and acetic acid bacteria international symposium [Esposizione]
Giudici, Paolo; Gullo, Maria; Solieri, Lisa; DE VERO, Luciana; Pulvirenti, Andrea; W., Ludwig; F., Barja; J., Bourgeois; J. M., Guillamon; A., Mas; P., Raspor
abstract

The key objective in planning the scientific programme was to ensure that the event would achieve the highest levels of scientific interest and topicality. Hence, the programme combines plenary session lectures, symposium sessions, as well as poster session.It would give a comprehensive overview of the most recent development in field of acetic acid bacteria and vinegar production and would surely stimulate fruitful discussions between attendees.One of the purposes of the meeting is also to create an international network for all the laboratories working in this field.


2004 - Cristallizzazione dell'aceto balsamico tradizionale [Articolo su rivista]
Giudici, Paolo; Pulvirenti, Andrea; DE VERO, Luciana; S., Landi; Gullo, Maria
abstract

Il processo di produzione dell’Aceto BalsamicoTradizionale (ABT) è complesso e dipendente davariabili fisiche, chimiche e biologiche. Durantel’invecchiamento si assiste ad una rilevanteconcentrazione dei soluti che possono dare luogoalla formazione di cristalli, in quantità tale daoccupare l’intero volume del contenitore. Nelpresente lavoro sono stati analizzati campioni diABT prelevati da barili contenenti il prodotto piùvecchio e che presentavano precipitato solido ingrande quantità. Il precipitato è risultato esserecomposto prevalentemente da glucosio ed inmisura minore da fruttosio, con un rapportofra i due zuccheri diverso da quello osservatonelle uve. La presenza di cristalli di zuccheronell’ABT è spiegato da due cause concomitanti:l’elevata concentrazione dei soluti e il rapportoglucosio/fruttosio a favore del primo. Lamaggiore presenza percentuale di glucosio èimputabile al tipo di lieviti che hanno condotto lafermentazione.Traditional Balsamic Vinegar (ABT) production iscomplex and influenced by physical, chemical,and biological parameters. During the aging,a remarkable concentration of soluble solids isobserved. These solids can make a great amountof crystals that fills the whole volume of thebottle. In the present work, the precipitatedsoluble solids from oldest ABT samples wereanalyzed.The precipitate is mainly glucose and in smalleramount is fructose. Therefore the ratio betweenthe two sugars is different from the one in thegrapes. The sugar crystals observed in aged ABTare explainable on the basis of two associatedcauses: the elevated soluble solids concentrationand the ratio glucose/fructose pro glucose. Thegreater amount of glucose is due to the yeastspecies that ferment the cooked must.


2004 - Etude De La Prédominance Des Levures Sélectionnée Dans Une Fermentation De Cave [Articolo su rivista]
Pulvirenti, Andrea; Castellari, L; DE PAOLA, M; Giudici, Paolo
abstract

Au cours d'une fermentation alcoolique, on considère qu'il y a prédominance de la souche de levure inoculée sur les levures indigènes quand le produit obtenu présente les caractéristiques d'un moût fermenté avec la souche pure, c'est-à-dire sans autres levures présentes. Dans la première partie de notre travail, nous avons voulu montrer que l'implantation de la souche inoculée peut être évaluée sur la base de la cohérence entre les résultats attendus et les caractéristiques du produit obtenu. Dans la seconde partie, nous avons évalué le rapport numérique entre les levures correspondant à la souche inoculée et la somme des levures indigènes, rapport qui nous permet d'affirmer la prédominance des premières.


2004 - I batteri acetici dell’Aceto Balsamico Tradizionale: caratterizzazione fenotipica e molecolare. [Relazione in Atti di Convegno]
M., Gullo; Pulvirenti, Andrea; DE VERO, Luciana; Giudici, Paolo
abstract


2004 - I lieviti dell’Aceto Balsamico Tradizionale dopo tre lustri. [Relazione in Atti di Convegno]
Solieri, Lisa; Pulvirenti, Andrea; A. R., Battagliola; M., Gullo; DE VERO, Luciana; Giudici, Paolo
abstract


2004 - I microrganismi dell'aceto balsamico. Atti convegno:"anche la tradizione va studiata" ricerche preliminari per l'individuazione di starter per l'Aceto Balsamico Tradizionale. [Curatela]
Giudici, Paolo; Gullo, Maria; Pulvirenti, Andrea; DE VERO, Luciana; Solieri, Lisa
abstract

Il ruolo e l'importanza dei microrganismi sulla qualità dell'aceto balsamico tradizionale.


2004 - Limits of r-DNA-NTS2 regions on the taxonomy of Saccharomyces genus [Relazione in Atti di Convegno]
Pulvirenti, Andrea; Solieri, Lisa; DE VERO, Luciana; Gullo, Maria; Giudici, Paolo
abstract

The aim of this work was to test different molecular techniques for determining which was the most effective in the identification of Saccharomyces cerevisiae strains. In particular, polymerase chain reaction--restriction fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer (ITS) regions and the nontranscribed spacer 2 (NTS2) region, sequencing of the D1/D2 domain, and electrophoretic karyotyping were applied to 123 yeast strains isolated from different sourdoughs and tentatively attributed to the species S. cerevisiae. All of the strains tested showed an identical PCR-RFLP pattern for the ITS regions, an identical nucleotide sequence of the D1/D2 domain, and the typical electrophoretic karyo type of S. cerevisiae. In contrast, 14 out of the 123 strains tested showed some polymorphism with BanI restriction analysis of the NTS2 region. Our results indicate that while the sequencing of the D1/D2 domain, the PCR-RFLP analysis of the ITS regions, and the electrophoretic karyotype can be employed successfully to identify S. cerevisiae strains, PCR-RFLP analysis of the NTS2 region does not allow a consistent and accurate grouping for S. cerevisiae strains. The fact that the NTS2 region of a small number of strains (8.78% of the total strains tested) is different from that of the other S. cerevisiae strains confirms that molecular methods should always be tested on a great number of strains.


2004 - Metodi indiretti per lo studio dei microrganismi dell’ABT. [Relazione in Atti di Convegno]
Pulvirenti, Andrea; Solieri, Lisa; Gullo, Maria; Giudici, Paolo
abstract

metodi coltura-indipendenti per la caratterizzazione della microflora dell'ABT


2004 - Occurrence and dominance of yeast species in sourdough [Articolo su rivista]
Pulvirenti, Andrea; Solieri, Lisa; Gullo, Maria; DE VERO, Luciana; Giudici, Paolo
abstract

Aims: The aim of this work is to identify the dominant yeast species in homemade sourdoughs. Methods and Results: PCR/restriction fragment length polymorphism analysis of internal transcribed spacer regions was used for the identification of isolates and the data were confirmed with phenotypic tests. The strains belonging to Saccharomyces cerevisiae were identified to strain level by analysis of inter-delta regions. Conclusion: This work shows that the dominant species in homemade sourdoughs can differ from each other. Saccharomyces cerevisiae was found to be the dominant species, followed by the Candida milleri, C. humilis, S. exiguus and Issatchenkia orientalis. The inter-delta regions of S. cerevisiae strains showed high polymorphism. Significance and Impact of the Study: Occurrence of single, non-Saccharomyces species and S. cerevisiae polymorphism in the yeast populations of sourdough samples.


2004 - Prevalence of the inoculated strain on the native yeast during wine fermentation [Relazione in Atti di Convegno]
Pulvirenti, Andrea; M., DE PAOLA; L., Castellari; Giudici, Paolo
abstract

International Congress on Yeasts. Yeasts in Sciences and Biotechnology - Prevalence of the inoculated strain on the native yeast during wine fermentation


2004 - Ricerche finalizzate alla tutela della tipicità dell'aceto balsamico di Reggio Emilia [Curatela]
Pulvirenti, Andrea
abstract

Nel Volume sono riportate le ricerche finalizzate alla tutela della tipicità dell'aceto balsamico di Reggio Emilia


2004 - Ricerche finalizzate alla tutela e tipicità dell'Aceto Balsamico Tradizionale di Reggio Emilia [Esposizione]
Giudici, Paolo; Gullo, Maria; DE VERO, Luciana; Pulvirenti, Andrea; U., Rangone
abstract

La valorizzazione e la tutela dell’Aceto Balsamico Tradizionale, al pari di qualsiasi altro prodotto alimentare tradizionale, non può prescindere dalla conoscenza delle ragioni che lo rendono così specifico e particolare. La ricerca scientifica, oltre ad essere lo strumento di conoscenza necessario alla comprensione della complessità dell’ABT, è anche veicolo di promozione, nel rispetto della chiarezza, dell’autenticità e dell’oggettività, senza ricorrere a spiegazioni aleatorie e fantasiose, che nulla hanno a che fare con la concretezza dell’ABT, frutto dell’acquisizione empirica della conoscenza.


2003 - Candida humilis - dominant species in sourdoughs for the production of durum wheat bran flour bread [Articolo su rivista]
Gullo, Maria; Romano, Ad; Pulvirenti, Andrea; Giudici, Paolo
abstract

Yeasts present in the sourdough that is generally used for the production of durum wheat bran flour bread were isolated and identified. Samples were taken during the rebuilding phase and at different intervals of time in order to monitor the population dynamics. The results obtained from the phenotypic studies were further confirmed by the molecular studies and enabled us to affirm that most of the strains, more than 95%, belong to the species Candida humilis. The dominance of C. humilis was steady in time. The isolations were carried out at sufficiently long intervals so that it was possible to ascertain that the conditions in which the sourdough is kept are fundamental to the microbiological stability of the dough.


2003 - Dominant yeast species in traditional sourdoughs [Abstract in Atti di Convegno]
Solieri, Lisa; Pulvirenti, Andrea; Gullo, Maria; Giudici, Paolo
abstract

Dominant yeast species in traditional sourdoughs


2003 - Estudio del predominio de las levaduras seleccionadas en la fermentacion en bodega [Monografia/Trattato scientifico]
Pulvirenti, Andrea; Giudici, Paolo
abstract

Estudio del predominio de las levaduras seleccionadas en la fermentacion en bodega por analiasi delle regioni interdelta


2003 - Identification des levures par techniques Moléculaires [Articolo su rivista]
Pulvirenti, Andrea; Giudici, Paolo
abstract

La taxonomie conventionnelle est fondée sur la modalité de reproduction sexuelle, si elle est présente, sur la morphologie des cellules et sur les caractéristiques physiologiques et biochimiques. Les tests sont souvent longs et laborieux et trois semaines sont souvent nécessaires pour obtenir les résultats complets. Ces dernières années, les temps demandés pour l'identification des levures ont été réduits par l'introduction de techniques moléculaires appliquées à l'étude des chromosomes entiers ou à l'analyse de domaines spécifiques ou une partie d'entre eux. Dans le présent travail, nous avons pris en considération l'emploi de l'électrophorèse en champ pulsé pour l'analyse du caryotype, le PCR/RFLP des régions NTS2 et ITS du rDNA, le séquençage de domaine D1/D2 du rDNA et, pour l'identification au niveau de la souche, le PCR des régions inter-δ et le RFLP de l'ADN mitochondrial.


2003 - In Vivo Interspecific Hybrids of Saccharomyces Species [Abstract in Atti di Convegno]
Pulvirenti, Andrea; Zambonelli, C.; Gullo, Maria; Giudici, Paolo
abstract

Costruction of in Vivo Saccharomyces Species Interspecific Hybrids


2003 - Microflora Y La Tecnología De Producción Del Vinagre Balsámico Tradicional. [Relazione in Atti di Convegno]
Giudici, Paolo; Gullo, Maria; Pulvirenti, Andrea; Solieri, Lisa; DE VERO, Luciana
abstract

Acetic acid bacteria of vinegar


2003 - Sorting of mitocondrial DNA and proteins in the progeny of Saccharomyces interspecific hybrids [Articolo su rivista]
De Vero, Luciana; Pulvirenti, Andrea; Gullo, Maria; Bonatti, Piera; Giudici, Paolo
abstract

The mitochondrial transmission to the progeny of interspecific hybrids Saccharomyces cerevisiae x Saccharomyces uvarum was investigated. It was observed that la progenie degli hybrids ha un solo tipo di mitocondri parentali come è stato osservato attraverso l’analisi RFLP dell’intero mtDNA. The mitochondrial proteins, invece, sono il risultato della ricombinazione del genoma dei due parentali. We suppose an important role of mitochondrial proteins in the process of mitochondrial transmission in yeast and hypothesize that inheritance of mtDNA may depend upon nuclear DNA genes. It seems to be probable that uniparental inheritance is independent on position of mitochondria in the buds of zygotes when the haploid cells fuse.


2003 - Studio molecolare e fenotipico di lieviti in paste acide tradizionali [Articolo su rivista]
Solieri, Lisa; DE VERO, Luciana; Gullo, Maria; Pulvirenti, Andrea
abstract

In questo lavoro è stato condotto uno studiodelle specie di lievito più rappresentatein impasti acidi tradizionali, preparatisenza l’aggiunta di lieviti starter. Tali speciesi contraddistinguono per importanzatecnologica in quanto numericamente prevalentie maggiormente responsabili delprocesso fermentativo. Le procedure diidentificazione adottate, basate su tecnichesia molecolari che fenotipiche, hannoconsentito di evidenziare una grande biodiversitànegli impasti acidi esaminati, ciascunodei quali rappresenta un unicuumsotto il profilo eumicetico.In this work a study on dominant yeastsspecies was carried out, using phenotypicand molecular techniques. These speciesare of technological interest since theyare the main responsible of the fermentationprocess. The obtained identificationsindicate that the examined home-madesourdoughs were characterized by a greatbiodiversity: each has got a distinguishingand unique yeast profile. The prevalenceof different yeast species in sourdoughssuggests to consider in additionto S. cerevisiae, also C. humilis, C. milleri,I. orientalis, and S. exiguus as speciessuitable for bread making.


2003 - Transmission of mt DNA and mitochondrial proteins in the progeny of Saccharomyces hybrids [Abstract in Atti di Convegno]
DE VERO, Luciana; Pulvirenti, Andrea; Gullo, Maria; Solieri, Lisa; Giudici, Paolo
abstract

Transmission of mt DNA and mitochondrial proteins in the progeny of Saccharomyces hybrids


2003 - Transmission of mtdna and mitochondrial proteins in the progeny of Saccharomyces interspecific hybrids. [Relazione in Atti di Convegno]
DE VERO, Luciana; Pulvirenti, Andrea; Gullo, Maria; Solieri, Lisa; Giudici, Paolo
abstract


2002 - A b-glucosidase positive strain of Saccharomyces cerevisiae isolated from grape must [Articolo su rivista]
C., Restuccia; Pulvirenti, Andrea; C., Caggia; Giudici, Paolo
abstract

A b-glucosidase positive strain of Saccharomyces cerevisiae isolated from grape must


2002 - Effects of a new packaging on the shelf-life of almond cookies [Articolo su rivista]
Lanza, C. M.; Tomaselli, F.; Pulvirenti, A.
abstract

To prolong the shelf-life of almond cookies the AA have proceeded to the packaging of the product (with and without sugar in surface) in polistyrene/poliethylene settled with flexible film in air and modified atmosphere. During the conservation period (50 days) physical-chemical, micro-biological, and sensorial controls have been carried out. The analysis of the Principal Components has underlined the difference between samples with and on the basis of the first 2 PC positively correlated with the sensorial parameters and negatively with the rheological softness. According to discriminant analysis the 62% of samples were correctly classified.


2002 - Horizontal transfer of genetic materials in Saccharomyces promoted by animals [Articolo su rivista]
Giudici, Paolo; Pulvirenti, Andrea; Todaro, Mary Antonio Donatello; Zambonelli, C.
abstract

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2002 - Interspecific hybridation by digestive tract of invertebrates as sorce of environmental biodiversity within the Saccaromyces cerevisiae. [Articolo su rivista]
Pulvirenti, Andrea; Zambonelli, C.; Todaro, Mary Antonio Donatello; Giudici, Paolo
abstract

Interspecific hybridation by digestive tract of invertebrates as sorce of environmental biodiversity within the Saccaromyces cerevisiae


2002 - Interspecific hybridisation by digestive tract of invertebrates as a source of environmental biodiversity within the Saccharomyces cerevisiae [Articolo su rivista]
Pulvirenti, Andrea; C., Zambonelli; Todaro, Mary Antonio Donatello; Giudici, Paolo
abstract

In order to verify whether animals can promote the formation of new yeast strains by increasing the chance of encounters between otherwise segregate spores within persistent asci, three invertebrate species representative of the marine, freshwater and terrestrial ecosystems were fed with two different diets composed of one or a combination of two strains of sporulated yeast and the egested material was analyzed for evidence. The digestive apparatus of the tested species is able to break open the asci wall (most probably by enzymatic action) produced in the species belonging to the Saccharomyces sensu stricto group without affecting the viability of the spores contained therein. The spores ejected with the fecal material have a high capacity for agglutination and, in the presence of favorable conditions, can germinate directly or conjugate to form hybrids. Hybrids between strains of the same species give rise to cultures in which the parent characteristics have new combinations. The interspecific hybrids Saccharomyces cerevisiae x Saccharomyces uvarum, although sterile, can propagate by asexual reproduction. These results support the hypothesis of horizontal transfer of genetic material between yeast species and the likelihood of hybrid formation in natural settings, and possibly shed light on the high biodiversity of yeast observed in nature.


2002 - Microbiologia degli aceti [Esposizione]
Giudici, Paolo; Pulvirenti, Andrea; Gullo, Maria
abstract

Il processo di produzione dell’Aceto Balsamico Tradizionale (ABT) è complesso ed è dipendente da variabili fisiche, chimiche e biologiche. Durante l’invecchiamento si assiste ad una rilevante concentrazione dei soluti tale per cui alcuni di essi possono precipitare dando luogo alla formazione di cristalli in quantità tale da essere visibili ad occhio nudo ed in alcuni casi possono occupare uno spazio corrispondente ad un terzo del prodotto. Nel presente lavoro abbiamo preso in considerazione campioni di ABT, prelevati dai barili contenente il prodotto più vecchio, che presentavano precipitato solido in grande quantità. Il precipitato è risultato essere rappresentato prevalentemente da glucosio ed in misura minore da fruttosio, mentre il rapporto fra i due zuccheri nel prodotto è risultato meno ampio, mantenendosi tuttavia lontano dal rapporto di circa 1/1 che si riscontra nelle uve. La ragione della formazione del precipitato nell’ABT invecchiato è spiegabile sulla base di due cause concomitanti: l’elevata concentrazione dei soluti, il rapporto glucosio/fruttosio spostato a favore del primo. La maggiore presenza percentuale di glucosio è imputabile al tipo di lieviti che hanno fermentato il mosto cotto base di partenza.


2002 - Molecular Methods for identification of wine yeasts [Capitolo/Saggio]
Giudici, Paolo; Pulvirenti, Andrea
abstract

Conventional taxonomy is based on the method of sexual reproduction, where is present, on the morphology of cells, and on physiological and biochemical features.


2002 - Uniparental Mitochondrial Inheritance in Saccaromyces spore coniugation. [Abstract in Atti di Convegno]
Pulvirenti, Andrea; Gullo, Maria; DE VERO, Luciana; Giudici, Paolo
abstract

Uniparental Mitochondrial Inheritance in Saccaromyces spore coniugation.


2001 - DNA fingerprinting methods used for identification of yeasts isolated from Sicilian sourdoughs [Articolo su rivista]
Pulvirenti, Andrea; C., Caggia; C., Restuccia; Gullo, Maria; Giudici, Paolo
abstract

The nature of sourdough yeast microflora was investigated. Samples were collected from different bakeries that do not use starter cultures, but rely on the traditional art of bread making. Pulsed field gel electrophoresis (PFGE), PCR/RFLP of the Non-Transcribed Spacer 2 (NTS2), the inter delta regions and the Internal Transcribed Spacer (ITS) of the ribosomal DNA (rDNA and partial sequencing of the 26S rDNA were used to identify and differentiate the yeast cultures isolated. In particular, PCR/RFLP of the ITS region, with restriction enzyme Hae III made it possible to differentiate Candida milleri from Candida humilis. A high degree of polymorphism was observed in the inter-delta regions of the strains belonging to the S. cerevisiae. species; contrary to what has been mentioned in literature, strains with the same profile are not necessarily identical. Saccharomyces cerevisiae, Saccharomyces cerevisiae old type anamensis, Issatchenkia orientalis, Candida mil[eri and Candida humilis were found to be the predominant species in Sicilian sourdough.


2001 - Identification of Pichia anomala isolated from yoghurt by RFLP of the ITS region [Articolo su rivista]
C., Caggia; C., Restuccia; Pulvirenti, Andrea; Giudici, Paolo
abstract

Several packs of swollen retailed plain and flavoured yoghurt were examined. The most commonly found species was Pichia anomala, identified both by physiological tests and RFLP analysis of the rDNA internal transcribed spacer (ITS) region. The isolated strains did not ferment lactose and were positive for galactose fermentation, confirming the hypothesis that galactose-fermenting yeast could be the cause of spoilage in yoghurt. (C) 2001 Elsevier Science B.V. All rights reserved.


2001 - Metodi molecolari per l’identificazione di lieviti isolati dalle paste acide siciliane [Capitolo/Saggio]
Pulvirenti, Andrea; Giudici, Paolo
abstract

Le paste acide sono impasti fermentati grazie alla presenza di una ricca floramicrobica costituita da batteri lattici e lieviti. Questi ultimi rivestono un ruolomolto importante, in quanto i prodotti primari del loro metabolismo energetico,anidride carbonica ed etanolo, determinano l’aumento di volume dell’impasto ela formazione della struttura porosa e alveolata della pasta, mentre alcuniprodotti secondari contribuiscono a determinare l’aroma tipico del pane. Lepaste acide sono costituite da farina, acqua e “lievito naturale”. Quest’ultimo èl’impasto della lavorazione precedente opportunamente rinnovato da impiegarecome coltura starter. Originariamente il lievito naturale era ottenutodall’acidificazione spontanea dell’impasto, dovuta alla contaminazione dellematerie prime e dagli attrezzi utilizzati. Il tempo necessario alla maturazionedegli impasti varia da qualche ora a tutta la notte precedente la lavorazionevera e propria. Invece con il termine “lievito madre” si intende la frazione diimpasto conservata in forma disidratata o congelata da impiegare come inoculoper la produzione di lievito naturale.In questo lavoro sono stati identificati, impiegando tecniche molecolari, ilieviti che caratterizzano gli impasti acidi siciliani. L’elettroforesi in campopulsato (PFGE), la PCR/RFLP delle regioni NTS2 (Non-Transcribed spacer2) eITS (Internal Transcribed Spacer) contenute nel DNA ribosomale (rDNA) edancora il parziale sequenziamento della subunità 26S del rDNA sono statiimpiegati per l’identificazione dei ceppi in esame. I risultati, di seguito esposti,mostrano la presenza di ceppi di lievito appartenenti alle specie Saccharomycescerevisiae, Saccharomyces anamensis, Issatchenkia orientalis, Candida milleri.


2000 - Effetti del confezionamento sulla conservabilità delle paste di mandorla [Articolo su rivista]
CARMELA M., Lanza; Filippo, Tomaselli; Pulvirenti, Andrea
abstract

Effetti del confezionamento sulla conservabilità delle paste di mandorla. Industrie Alimentari.


2000 - Inheritance of mitochondrial DNA in interspecific Saccharomyces hybrids [Articolo su rivista]
Pulvirenti, Andrea; C., Caggia; C., Restuccia; Giudici, Paolo; C., Zambonelli
abstract

Spore conjugation between Saccharomyces cerevisiae and S. uvarum strains yields hybrids which have mitochondria of only a parental type: either of S. cerevisiae or S. uvarum. The peculiarity of examined hybrids is due to the homogeneity of their clone populations that are consisting of cells, all characterised by the same mitochondrial type. It seems to be probable to suppose an incompatibility of parental mitochondrial DNA (mtDNA).


2000 - Rapid differentiation of the closely related Kluyveromyces lactis var. lactis and K. marxianus strains isolated from dairy products using selective medium and PCR/RFLP of the rDNA non trascribed spacer 2 [Articolo su rivista]
NGUYEN H., Vang; Pulvirenti, Andrea; Claude, Gaillarden
abstract

Rapid differentiation of the closely related Kluyveromyces lactis var. lactis and K. marxianus strains isolated from dairy products using selective medium and PCR/RFLP of the rDNA non trascribed spacer 2


2000 - Saccharomyces uvarum, a proper species within Saccharomyces sensu stricto [Articolo su rivista]
Pulvirenti, Andrea; Hv, Nguyen; C., Caggia; Giudici, Paolo; S., Rainieri; C., Zambonelli
abstract

Saccharomyces uvarum is proposed as a proper species within the complex Saccharomyces sensu stricto. Molecular characteristics including the similarity of the restriction profile of the non-transcribed spacer 2 (NTS2) and of the D1/D2 sequences of the rDNA, as well as other genotypic and phenotypic characteristics confirm that this group of strains is highly homogeneous and distinguishable from other species of the Saccharomyces sensu stricto group. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.


2000 - Studio sulla conservabilità delle brioche da gelato. [Articolo su rivista]
CARMELA M., Lanza; Filippo, Tomaselli; Giuseppe, Muratore; Pulvirenti, Andrea
abstract

Studio sulla conservabilità delle brioche da gelato


1999 - Cryotolerant Saccharomyces strains and spoilage o f refrigerated must [Articolo su rivista]
Giudici, Paolo; Caggia, C.; Pulvirenti, Andrea
abstract

This study regards yeasts isolated from refrigerated musts with patent ongoing fermentation. The isolates belonged to the genus Saccharomyces sensu stricto with an optimal growth temperature less than 30 °C. Karyotype analysis highlighted the constant presence of two well-defined trait bands in the low molecular weight between 225 and 375 kb that distinguished them unequivocally from normal Saccharomyces cerevisiae strains. The isolated strains individually reinoculated in sterile must at 25 °C, produce complete fermentation. Analysis of minor fermentation compounds revealed other differences between these strains and Saccharomyces cerevisiae strains. These strains are responsible for the spoilage of fermenting refrigerated must. Phenotype traits, combined with karyotype, suggest that they should not be assigned to Saccharomyces cerevisiae but more reasonably to Saccharomyces bayanus uvarum type.


1999 - Cryotolerant Saccharomyces strains and spoilage of refrigerated musts [Articolo su rivista]
Giudici, Paolo; C., Caggia; Pulvirenti, Andrea; C., Restuccia
abstract

This study regards yeasts isolated from refrigerated musts with patent ongoing fermentation. The isolates belonged to the genus Saccharomyces sensu stricto with an optimal growth temperature less than 30 degrees C. Karyotype analysis highlighted the constant presence of two well-defined trait bands in the low molecular weight between 225 and 375 kb that distinguished them unequivocally from normal Saccharomyces cerevisiae strains. The isolated strains individually reinoculated in sterile must at 25 degrees C, produce complete fermentation. Analysis of minor fermentation compounds revealed other differences between these strains and Saccharomyces cerevisiae strains. These strains are responsible for the spoilage of fermenting refrigerated must. Phenotype traits, combined with karyotype, suggest that they should not be assigned to Saccharomyces cerevisiae but more reasonably to Saccharomyces bayanus uvarum type.


1999 - Identification of yeasts isolated from spoiled yoghurt [Monografia/Trattato scientifico]
Caggia, C.; Restuccia, C.; Pulvirenti, Andrea; Giudici, Paolo
abstract

everal packs of swollen retailed plain and flavoured yoghurt were examined. The most commonly found species was Pichia anomala, identified both by physiological tests and RFLP analysis of the rDNA internal transcribed spacer (ITS) region. The isolated strains did not ferment lactose and were positive for galactose fermentation, confirming the hypothesis that galactose-fermenting yeast could be the cause of spoilage in yoghurt.


1999 - Saccharomyces uvarum, a distinct group within Saccharomyces sensu stricto [Articolo su rivista]
S., Rainieri; C., Zambonelli; E., JOHON HALLSWORT; Pulvirenti, Andrea; Giudici, Paolo
abstract

Saccharomyces uvarum, a distinct group within Saccharomyces sensu stricto


1998 - Electroforetic profiles of hybrids between cryotolerant and non-cryotolerant Saccharomyces strains [Articolo su rivista]
Giudici, Paolo; C., Caggia; Pulvirenti, Andrea; C., Zambonelli; S., Rainieri
abstract

The chromosomal DNAs of cryotolerant Saccharomyces bayanus, non-cryotolerant Saccharomyces cerevisiae strains and their intra and interspecific hybrids were separated by pulsed field electrophoresis (PFGE). The cryotolerant and non-cryotolerant strains gave distinctly different electrophoretic profiles. The hybrids cryotolerant x cryotolerant and non-cryotolerant x non-cryotolerant were fertile and they gave the same electrophoretic karyotype as the respective parents. The cryotolerant x non-cryotolerant hybrids were sterile and gave electrophoretic karyotypes which showed both the bands the parents have in common and those they do not share.


1998 - Electrophoretic Profile of Thermotolerant Saccaromyces cerevisiae [Articolo su rivista]
S., Rainieri; Pulvirenti, Andrea; C., Caggia; Giudici, Paolo
abstract

Electrophoretic Profile of Thermotolerant Saccaromyces cerevisiae


1998 - Karyotiping of Saccharomyces strains with different temperature profiles [Articolo su rivista]
Giudici, Paolo; C., Caggia; Pulvirenti, Andrea; S., Ranieri
abstract

This study examined the, the fermentation performance and the optimum growth temperature (Topt) of 28yeast strains all identified as species belonging to Saccharomyces sensu stricto. The strainswere isolated from fermented musts, which had not been inoculated, at twotemperature ranges: 20–40 °C and approximately 0–6 °C. The results demonstrated acorrelation between the Topt and the chromosome organization. In particular, strains withTopt of less than 30 °C showed only two bands in the region between 365 and 225 kb,while those with a Topt greater than 30 °C had three bands in this size range. From ataxonomic viewpoint, the Topt is a better indicator for the Saccharomyces sp. than theceiling temperature of 37 °C currently used to differentiate cryotolerantSaccharomyces bayanus and S. pastorianus from non-cryotolerant S. cerevisiae and S.paradoxus strains.


1998 - Pulsed field gel electrophoresis: optimisation of the running conditions to improve the differentiation of Saccharomyces cerevisiae and Saccharomyces bayanus strains [Abstract in Atti di Convegno]
Pulvirenti, Andrea; C., Caggia; S., Rainieri; Giudici, Paolo
abstract

Pulsed field gel electrophoresis: optimisation of the running conditions to improve the differentiation of Saccharomyces cerevisiae and Saccharomyces bayanus strains


1997 - I batteri lattici del pecorino siciliano [Articolo su rivista]
Daniela, Migliorisi; Cinzia, Caggia; Pulvirenti, Andrea; Giudici, Paolo
abstract

I batteri lattici del pecorino siciliano


1997 - La sicurezza microbiologica nella preparazione casalinga delle olive verdi al naturale [Articolo su rivista]
Giudici, Paolo; MARIA CONCETTA, Strano; Pulvirenti, Andrea
abstract

sicurezza microbiologica nella preparazione casalinga delle olive verdi al naturale


1997 - La sicurezza microbiologica nella preparazione casalinga delle olive verdi al naturale siciliane. [Articolo su rivista]
Giudici, Paolo; M. C., Strano; Pulvirenti, Andrea
abstract

La sicurezza microbiologica nella preparazione casalinga delle olive verdi al naturale siciliane.


1997 - L’acido gluconico e l’acido lattico come indici di qualità microbiologica dei succhi d’arancia [Articolo su rivista]
Giudici, Paolo; Pulvirenti, Andrea; C., Caggia
abstract

L’acido gluconico e l’acido lattico come indici di qualità microbiologica dei succhi d’arancia