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"Pellegrini, Michela"
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New Methodologies as Opportunities in the Study of Bacterial Biofilms, Including Food-Related Applications
by
Wang, Zhe
,
Bianco, Vittorio
,
Coppola, Francesca
in
Adaptability
,
Antibiotics
,
Antimicrobial agents
2025
Traditional food technologies, while essential, often face limitations in sensitivity, real-time detection, and adaptability to complex biological systems such as microbial biofilms. These constraints have created a growing demand for more advanced, precise, and non-invasive tools to ensure food safety and quality. In response to these challenges, cross-disciplinary technological integration has opened new opportunities for the food industry and public health, leveraging methods originally developed in other scientific fields. Although their industrial-scale implementation is still evolving, their application in research and pilot settings has already significantly improved our ability to detect and control biofilms, thereby strengthening food safety protocols. Advanced analytical techniques, the identification of pathogenic species and their virulence markers, and the screening of “natural” antimicrobial compounds can now be conceptualized as interconnected elements within a virtual framework centered on “food” and “biofilm”. In this short review, starting from the basic concepts of biofilm and associated microorganisms, we highlight a selection of emerging analytical approaches—from optical methods, microfluidics, Atomic Force Microscopy (AFM), and biospeckle techniques to molecular strategies like CRISPR, qPCR, and NGS, and the use of organoids. Initially conceived for biomedical and biotechnological applications, these tools have recently demonstrated their value in food science by enhancing our understanding of biofilm behavior and supporting the discovery of novel anti-biofilm strategies.
Journal Article
Alcohol or No Alcohol in Wine: Half a Century of Debate
by
Testa, Bruno
,
Coppola, Francesca
,
Iorizzo, Massimo
in
Alcohol abuse
,
Alcohol free
,
Alcoholic beverages
2025
Alcoholic beverages have been consumed for centuries in different countries around the world. Today, we know that the ethanol they contain is associated with significant health risks, especially in the case of abuse, in individuals with special health conditions, and in pregnant women. However, over the years, awareness has grown that wine, especially red wine, has a beneficial effect on human health due to the powerful effect of the antioxidant substances it contains, known under the generic term of polyphenols. The main concern remains around the ethanol content of wine and its effects on health. After fifty years of research and studies, the debate is still open, with conflicting indications about the positive effect of moderate wine consumption in the context of a balanced diet and the toxic effect of ethanol even in low doses. In this disputed area, the market for low-alcohol and alcohol-free wines has found its place in the last decade, creating a new opportunity for the global wine trade. These new types of wine are going to open a new chapter in the history of wine. In this review, we have summarised the main aspects of the health implications of wine consumption considering scientific evidence from the last 50 years, including low-alcohol and dealcoholised wine.
Journal Article
Evaluation of Total Volatile Basic Nitrogen, Formaldehyde, and Formic Acid as Markers to Define the Acceptability of Farmed Sea Bass and Sea Bream Stored Under Vacuum (VP) or in Modified-Atmosphere Packaging (MAP) at 4 ± 2 °C
2025
The aim of this work was to propose an objective parameter to define the acceptability of fresh sea bass and sea bream among concentrations of formaldehyde, formic acid, and TVB-N. As indicated, TVB-N appeared to be the most appropriate index. The formaldehyde value cannot be used because it increased until day 6 and then decreased because it was transformed into formic acid. The decrease was observed at all of the times tested. Nevertheless, formic acid also cannot be considered as a valid parameter because in both of the tested fish, it reached values of less than 7.2 mg/kg at 15 days of storage, even though the sensorial analysis indicated the loss of acceptability. In addition, this value is 4-7 times lower than the concentration present in other fresh fish. Consequently, TVB-N represents the only parameter of interest for defining acceptability for both fish species and it can be accepted as the freshness index. Considering the results of the microbial, physico-chemical, and sensory analysis, a level of TVB-N less or equal to 35 mg N/100 g of product was observed up to 12 days of storage. Then, at 15 days, TVB-N reached values over 40 mg N/100 g and both the fish were no longer acceptable, as demonstrated by sensory analysis. For this reason, this value can also be proposed as the limit of freshness for sea bass and sea bream, stored either in VP or in MAP at 4 ± 2 °C. Considering the microbial, physico-chemical, and sensorial analysis of both fish species, a shelf-life limit of 12 days was proposed.
Journal Article
Chalk Yeasts Cause Gluten-Free Bread Spoilage
by
Montanari, Chiara
,
Barbieri, Federica
,
Iacumin, Lucilla
in
Acetic acid
,
Alcohols
,
Amino acids
2025
Four different yeast strains were isolated from industrial gluten-free bread (GFB) purchased from a local supermarket. These strains, including Hyphopichia burtonii, Wickerhamomyces anomalus, Saccharomycopsis fibuligera, and Cyberlindnera fabianii, are responsible for spoilage, which consists of white powdery and filamentous colonies due to the fragmentation of hyphae into short-length fragments (dust-type spots) that is typical of the spoilage produced by chalk yeasts. The isolated strains were identified using genomic analysis. Among them, C. fabianii was also isolated, which is a rare ascomycetous opportunistic yeast species with low virulence attributes, uncommonly implicated in bread spoilage. The yeast growth was studied in vitro on Malt Extract Agar (MEA) at two temperatures (20 and 25 °C) and at different Aws (from 0.99 to 0.90). It was inferred that the temperature did not influence the growth. On the contrary, different Aws reduced the growth, but all the yeast strains could grow until a minimum Aw of about 0.90. Different preservatives (ethanol, hop extract, and sorbic and propionic acids) were used to prevent the growth. In MEA, the growth was reduced but not inhibited. In addition, the vapor-phase antimicrobial activity of different preservatives such as ethanol and hop extract was studied in MEA. Both preservatives completely inhibited the yeast growth either at 20 or at 25 °C. Both preservatives were found in GFB slices. Contrary to hop extract, 2% (v/w) ethanol completely inhibited all the strains. The spoilage was also confirmed by the presence of various compounds typically present in yeasts, derived from sugar fermentation and amino acid degradation. These compounds included alcohols, ketones, organic acids, and esters, and they were identified at higher concentrations in the spoiled samples than in the unspoiled samples. The concentration of acetic acid was low only in the spoiled samples, as this compound was consumed by yeasts, which are predominately present in the spoiled samples, to produce acetate esters.
Journal Article
Diversity of Lactiplantibacillus plantarum in Wild Fermented Food Niches
by
Gagliardi, Roberto
,
Iarusso, Ilenia
,
Succi, Mariantonietta
in
Adaptation
,
Bacteria
,
Bacteriocins
2025
This study aimed to explore the genetic and functional diversity of Lactiplantibacillus plantarum (Lpb. plantarum) strains from wild fermented foods to identify traits that are useful for food innovation. The growing demand for clean-label, plant-based, and functionally enriched fermented foods exposes the limitations of current industrial fermentation practices, which rely on standardized lactic acid bacteria (LAB) strains with limited metabolic plasticity. This constraint hinders the development of new food formulations and the replacement of conventional additives. To address this gap, 343 LAB strains were analyzed, including 69 Lpb plantarum strains, isolated from five minimally processed, spontaneously fermented matrices: fermented millet, kombucha, and sourdough (plant-based), wild fermented mountain milk, and natural whey starter (animal-based). Whole-genome sequencing was performed to assess phylogenetic relationships and to annotate genes encoding carbohydrate-active enzymes (CAZymes) and antimicrobial compounds. The results revealed a marked strain-level diversity. Glycoside hydrolase (GH) families GH13 and GH1 were widely distributed, while GH25 and GH32 showed variable presence across clusters. Strains grouped into clusters enriched with plant-based isolates exhibited distinct CAZyme profiles adapted to complex carbohydrates. Clusters with animal-based strains exhibited a broader gene repertoire related to bacteriocin biosynthesis. These findings highlight the untapped potential of wild fermented food environments as reservoirs of Lpb. plantarum with unique genomic traits. Harnessing this diversity can expand the functional capabilities of starter cultures, promoting more sustainable, adaptive, and innovative fermentation systems. This study underscores the strategic value of underexploited microbial niches in meeting the evolving demands of modern food production.
Journal Article
Hygienic Quality of Air-Packed and Refrigerated or Frozen Stored Döner Kebab and Evaluation of the Growth of Intentionally Inoculated Listeria monocytogenes
by
Coppola, Francesca
,
Iacumin, Lucilla
,
Ferluga, Giada
in
Air quality
,
Analysis
,
Biodegradation
2025
Döner kebab, a meat product of Middle-Eastern origin, has gained significant popularity and is now widely consumed across Europe. The recipe varies depending on the area, with beef, turkey, lamb, or chicken being used as main ingredients. The aim of this work was to assess the hygienic-sanitary quality of raw and cooked döner kebabs stored at 4 ± 2 °C for 10 days and at 8 ± 2 °C for the next 20 days or frozen (−18 °C) for one month. One additional aim was to determine the potential growth of Listeria monocytogenes intentionally inoculated in cooked döner kebab during storage at 4 ± 2 °C or freezing. The concentration of Total Viable Count (TVC) and the Enterobacteriaceae of the 100 samples of raw döner kebab were less than 7 log CFU/g and 4 log CFU/g, respectively. Consequently, the samples can be considered acceptable and similar to traditional raw meat. The cooked döner kebab can be considered safe for a period of 30 days, especially from a microbiological point of view, when stored under refrigerated conditions, also taking into account possible thermal abuse. Coagulase Positive Cocci (CPC), Clostridium H2S+, Salmonella spp., and Listeria monocytogenes were never found in any of the samples. After 30 days, the TVC was at the level of 6 log CFU/g and Enterobacteriaceae at less than 4 log CFU/g. The main concern was related to microbial or tissue activity, resulting in an increase in total volatile basic nitrogen (TVB-N) content. However, in the cooked samples, the TVB-N content remained below 40 mg N/100 g at the end of the shelf-life period (32.5 mg N/100 g), which is still considered an acceptable value. In addition, the level of Malondialdehyde (MDA) was found to be within acceptable limits, with a reading of 1.4 nmol/g attained after 30 days. The same product, when frozen and stored at −18 °C, can be considered stable for a minimum of 6 months, both from a microbiological and a physico-chemical point of view. No microbial growth was observed. The TVB-N and the MDA levels increased, but after 6 months, their levels were still acceptable, with values of 19.1 mg N/100 g and 1.2 nmol/g, respectively. These observations demonstrate low protein degradation and lipid oxidation during the shelf-life period. The challenge test showed that Listeria monocytogenes did not grow in döner kebab either when stored at 4 ± 2 °C for 10 days and 8 ± 2 °C for 20 days or when stored at −18 °C for 6 months. The concentration of L. monocytogenes was found to be 5.4 log CFU/g in the refrigerated products and 4.9 log CFU/g in the frozen products. At the end of the shelf-life period, the L. monocytogenes load in both products was lower than the initial concentration that had been added. Finally, the use of air-packaging has been proven to be beneficial to the preservation of the product and maintained its microbiological and physico-chemical properties intact. Despite these good results, future directions could be to investigate different plastic films and packaging such as Modified Atmosphere (MAP), Vacuum (VP), and Sous Vide packaging (SVP).
Journal Article
Evaluation of Different Techniques, including Modified Atmosphere, under Vacuum Packaging, Washing, and Latilactobacillus sakei as a Bioprotective Agent, to Increase the Shelf-Life of Fresh Gutted Sea Bass (Dicentrarchus labrax) and Sea Bream (Sparus aurata) Stored at 6 ± 2 °C
2022
Fish meat is very perishable because of indigenous and microbial enzymes, which determine spoilage and shelf life. The deterioration processes, which lead to an important, sequential, and progressive modification of the initial state of freshness, are fast and depend on rearing, harvesting, slaughtering, handling, and storage conditions. Usually, the shelf life of gutted fish stored at 4 ± 2 °C under vacuum packaging (VP—1.0 bar) and modified atmosphere packaging (MAP, 70% N2, <1% O2, 30% CO2) is approximately 9 days. The aim of this work was to improve the shelf life and preserve the microbiological and sensory quality of farmed gutted sea bass (Dicentrarchus labrax) and sea bream (Sparus aurata) using different methods, including VP, MAP, and bioprotective culture containing Latilactobacillus sakei, until 12–14 days. Microbiological, physicochemical, and sensory quality indices were monitored to confirm the effectiveness of biopreservation on product quality during proper refrigeration (4 ± 2 °C) or abuse (6 ± 2 °C, simulating supermarkets and consumer fridges) storage period. Considering the quality indexes represented by Enterobacteriaceae, total volatile nitrogen (TVB-N), and malonaldehyde concentrations (TBARS) and the sensorial analysis, the VP samples were more acceptable than the MAP fish, even though the shelf-life of the VP and MAP fish was similar at about 12 days. The second phase of the work was to evaluate the shelf-life of both VP fish stored at 6 ± 2 °C, which simulates the normal abuse temperature of supermarkets or consumer fridges. Data confirmed the previous results and demonstrated, despite the abuse temperature of storage, a shelf-life of about 12 days. Finally, the third phase consisted of prolonging the shelf life until 14 days of storage at 6 ± 2 °C by washing the gutted sea bass and sea bream in a suspension of bioprotective starter (7 log CFU/mL) with or without the addition of dextrose (0.1%) and by VP packaging. The bioprotective culture reduced the growth of spoilage microorganisms. Consequently, the total volatile nitrogen (TVB-N) concentration in both fish species was low (<35 mg N/100 g). Nonprofessional and untrained evaluators confirmed the acceptability of the inoculated samples by sensorial analysis.
Journal Article
Use of Inulin and Pumpkin Oil in the Manufacture of High-Quality Mortadella-Style Sausage from Buffalo Meat
by
Grazia, Luigi
,
Iarusso, Ilenia
,
Coppola, Francesca
in
Animal fat
,
Antimicrobial activity
,
Antimicrobial agents
2025
The growing demand for healthier meat products has driven the reformulation of processed meats to reduce saturated fat while preserving sensory and technological attributes. Buffalo meat (Bubalus bubalis), with its high protein content, low intramuscular fat, and favorable fatty acid profile, offers a promising base for healthier formulations. However, its fat content may compromise texture, juiciness, and flavor, necessitating strategies to optimize product quality. This study investigated the effects of replacing pork fat with inulin and pumpkin seed oil in a cooked buffalo meat product, focusing on compositional, oxidative, microbiological, and sensory parameters. Two plant-based ingredients were selected: inulin from chicory, used as a fat mimic due to its gel-forming ability, and pumpkin seed oil, a structural analog with antimicrobial activity. Preliminary trials identified optimal concentrations for balancing technological and functional performance. A 2% inclusion of pumpkin seed oil, exceeding its in vitro MIC (0.4–1.5%), ensured effectiveness in the food matrix. Reformulated products exhibited significantly reduced fat (p < 0.05), enhanced fiber, and a lipid profile rich in polyunsaturated fatty acids (>45%), qualifying for European Union health claims. Oxidative stability improved (p < 0.01), and sensory analysis revealed enhanced aroma complexity, with nutty and roasted notes. Microbiological assessments confirmed a protective effect against spoilage bacteria. These results support the development of a nutritionally improved, microbiologically safer cooked product, such as mortadella-style sausage, while also offering strategies for broader innovation in reformulating functional meat products.
Journal Article
Microbial Spoilage of Traditional Goose Sausages Produced in a Northern Region of Italy
2023
Recently, during the ripening of goose sausage, a defect consisting of ammonia and vinegar smell was noticed. The producer of the craft facility, located in Lombardia, a Northern region of Italy, asked us to identify the cause of that defect. Therefore, this study aimed to identify the potential responsible agents for the spoilage of this lot of goose sausages. Spoilage was first detected by sensory analysis using the “needle probing” technique; however, the spoiled sausages were not marketable due to the high ammonia and vinegar smell. The added starter culture did not limit or inhibit the spoilage microorganisms, which were represented by Levilactobacillus brevis, the predominant species, and by Enterococcus faecalis and E. faecium. These microorganisms grew during ripening and produced a large amount of biogenic amines, which could represent a risk for consumers. Furthermore, Lev. brevis, being a heterofermentative lactic acid bacteria (LAB), also produced ethanol, acetic acid, and a variation in the sausage colour. The production of biogenic amines was confirmed in vitro. Furthermore, as observed in a previous study, the second cause of spoilage can be attributed to moulds which grew during ripening; both the isolated strains, Penicillium nalgiovense, added as a starter culture, and P. lanosocoeruleum, present as an environmental contaminant, grew between the meat and casing, producing a large amount of total volatile nitrogen, responsible for the ammonia smell perceived in the ripening area and in the sausages. This is the first description of Levilactobacillus brevis predominance in spoiled goose sausage.
Journal Article
Microbial Characterization of Retail Cocoa Powders and Chocolate Bars of Five Brands Sold in Italian Supermarkets
by
Iacumin, Lucilla
,
Comi, Giuseppe
,
Colautti, Andrea
in
Bacillus (bacteria)
,
Bacteria
,
Chemical properties
2022
A microbial characterization of cocoa powder and chocolate bars of three batches of five different brands sold in Italian markets was performed. The results showed a variable microbial population consisting of mesophilic and thermophilic spore formation in both types of products. The chocolate bars were also contaminated with molds of environmental origin. Bacillus spp. and Geobacillus spp. were found in both products. The chocolate bars were also contaminated by molds belonging to the genera Penicillium and Cladosporium. The sporogenous strains mainly originate from the raw materials, i.e., cocoa beans, as the heat treatments involved (roasting of the beans and conching of the chocolate) are not sufficient to reach commercial sterility. Furthermore, the identified spore-forming species have often been isolated from cocoa beans. The molds isolated from chocolate seem to have an origin strictly linked to the final phases of production (environment and packaging). However, the level of contaminants is limited (<2 log CFU/g); the molds do not develop in both products due to their low Aw (<0.6) and do not affect the safety of the products. However, a case of mold development in chocolate bars was observed. Among the isolated molds, only Penicillium lanosocoeruleum demonstrated a high xero-tolerance and grew under some conditions on chocolate bars. Its growth could be explained by a cocoa butter bloom accompanied by the presence of humidity originating from the bloom or acquired during packaging.
Journal Article