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"Comi, Giuseppe"
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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
Effect of a Debaryomyces hansenii and Lactobacillus buchneri Starter Culture on Aspergillus westerdijkiae Ochratoxin A Production and Growth during the Manufacture of Short Seasoned Dry-Cured Ham
2020
Recently, specific dry-cured hams have started to be produced in San Daniele and Parma areas. The ingredients are similar to protected denomination of origin (PDO) produced in San Daniele or Parma areas, and include pork leg, coming from pigs bred in the Italian peninsula, salt and spices. However, these specific new products cannot be marked as a PDO, either San Daniele or Parma dry cured ham, because they are seasoned for 6 months, and the mark PDO is given only to products seasoned over 13 months. Consequently, these products are called short-seasoned dry-cured ham (SSDCH) and are not branded PDO. During their seasoning period, particularly from the first drying until the end of the seasoning period, many molds, including Eurotium spp. and Penicillium spp., can grow on the surface and work together with other molds and tissue enzymes to produce a unique aroma. Both of these strains typically predominate over other molds. However, molds producing ochratoxins, such as Aspergillus ochraceus and Penicillium nordicum, can simultaneously grow and produce ochratoxin A (OTA). Consequently, these dry-cured hams may represent a potential health risk for consumers. Recently, Aspergillus westerdijkiae has been isolated from SSDCHs, which could represent a potential problem for consumers. Therefore, the aim of this study was to inhibit A. westerdijkiae using Debaryomyces hansenii or Lactobacillus buchneri or a mix of both microorganisms. Six D. hansenii and six L. buchneri strains were tested in vitro for their ability to inhibit A. westerdijkiae. The strains D. hansenii (DIAL)1 and L. buchneri (Lb)4 demonstrated the highest inhibitory activity and were selected for in situ tests. The strains were inoculated or co-inoculated on fresh pork legs for SSDCH production with OTA-producing A. westerdijkiae prior to the first drying and seasoning. At the end of seasoning (six months), OTA was not detected in the SSDCH treated with both microorganisms and their combination. Because both strains did not adversely affect the SSDCH odor or flavor, the combination of these strains are proposed for use as starters to inhibit OTA-producing A. westerdijkiae.
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
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
Cyberlindnera fabianii, an Uncommon Yeast Responsible for Gluten Bread Spoilage
2024
A single strain of yeast was isolated from industrial gluten bread (GB) purchased from a local supermarket. This strain is responsible for spoilage consisting of white powdery and filamentous colonies due to the fragmentation of hyphae into short lengths (dust-type spots), similar to the spoilage produced by chalk yeasts such as Hyphopichia burtonii, Wickerhamomyces anomalus and Saccharomycopsis fibuligera. The isolated strains were identified initially by traditional methods as Wickerhamomyces anomalus, but with genomic analysis, they were definitively identified as Cyberlindnera fabianii, a rare ascomycetous opportunistic yeast species with low virulence attributes, uncommonly implicated in bread spoilage. However, these results demonstrate that this strain is phenotypically similar to Wi. anomalus. Cy. fabianii grew in GB because of its physicochemical characteristics which included pH 5.34, Aw 0.97 and a moisture of about 50.36. This spoilage was also confirmed by the presence of various compounds typical of yeasts, derived from sugar fermentation and amino acid degradation. These compounds included alcohols (ethanol, 1-propanol, isobutyl alcohol, isoamyl alcohol and n-amyl alcohol), organic acids (acetic and pentanoic acids) and esters (Ethylacetate, n-propil acetate, Ethylbutirrate, Isoamylacetate and Ethylpentanoate), identified in higher concentrations in the spoiled samples than in the unspoiled samples. The concentration of acetic acid was lower only in the spoiled samples, but this effect may be due to the consumption of this compound to produce acetate esters, which predominate in the spoiled samples.
Journal Article
Phage Inactivation of Listeria monocytogenes on San Daniele Dry-Cured Ham and Elimination of Biofilms from Equipment and Working Environments
2016
The anti-listerial activity of generally recognized as safe (GRAS) bacteriophage Listex P100 (phage P100) was demonstrated in broths and on the surface of slices of dry-cured ham against 5 strains or serotypes (i.e., Scott A, 1/2a, 1/2b, and 4b) of Listeria monocytogenes. In a broth model system, phage P100 at a concentration equal to or greater than 7 log PFU/mL completely inhibited 2 log CFU/cm2 or 3 log CFU/cm2 of L. monocytogenes growth at 30 °C. The temperature (4, 10, 20 °C) seemed to influence P100 activity; the best results were obtained at 4 °C. On dry-cured ham slices, a P100 concentration ranging from 5 to 8 log PFU/cm2 was required to obtain a significant reduction in L. monocytogenes. At 4, 10, and 20 °C, an inoculum of 8 log PFU/cm2 was required to completely eliminate 2 log L. monocytogenes/cm2 and to reach the absence in 25 g product according to USA food law. Conversely, it was impossible to completely eradicate L. monocytogenes with an inoculum of approximately of 3.0 and 4.0 log CFU/cm2 and with a P100 inoculum ranging from 1 to 7 log PFU/cm2. P100 remained stable on dry-cured ham slices over a 14-day storage period, with only a marginal loss of 0.2 log PFU/cm2 from an initial phage treatment of approximately 8 log PFU/cm2. Moreover, phage P100 eliminated free L. monocytogenes cells and biofilms on the machinery surfaces used for dry-cured ham production. These findings demonstrate that the GRAS bacteriophage Listex P100 at level of 8 log PFU/cm2 is listericidal and useful for reducing the L. monocytogenes concentration or eradicating the bacteria from dry-cured ham.
Journal Article
Assessment of Aerobic and Respiratory Growth in the Lactobacillus casei Group
by
Ricciardi, Annamaria
,
Parente, Eugenio
,
Zotta, Teresa
in
Adaptation, Physiological
,
Aerobic conditions
,
Aerobiosis
2014
One hundred eighty four strains belonging to the species Lactobacillus casei, L. paracasei and L. rhamnosus were screened for their ability to grow under aerobic conditions, in media containing heme and menaquinone and/or compounds generating reactive oxygen species (ROS), in order to identify respiratory and oxygen-tolerant phenotypes. Most strains were able to cope with aerobic conditions and for many strains aerobic growth and heme or heme/menaquinone supplementation increased biomass production compared to anaerobic cultivation. Only four L. casei strains showed a catalase-like activity under anaerobic, aerobic and respiratory conditions and were able to survive in presence of H2O2 (1 mM). Almost all L. casei and L. paracasei strains tolerated menadione (0.2 mM) and most tolerated pyrogallol (50 mM), while L. rhamnosus was usually resistant only to the latter compound. This is the first study in which an extensive screening of oxygen and oxidative stress tolerance of members of the L. casei group has been carried out. Results allowed the selection of strains showing the typical traits of aerobic and respiratory metabolism (increased pH and biomass under aerobic or respiratory conditions) and unique oxidative stress response properties. Aerobic growth and respiration may confer technological and physiological advantages in the L. casei group and oxygen-tolerant phenotypes could be exploited in several food industry applications.
Journal Article
Listeria monocytogenes Survey in Cubed Cooked Ham Packaged in Modified Atmosphere and Bioprotective Effect of Selected Lactic Acid Bacteria
2020
The aim of this work was to study the presence of Listeria monocytogenes, as well as the potential activity of two bioprotective cultures (Lyocarni BOX-74 and Lyocarni BOX-57), versus a mix of three L. monocytogenes strains that were intentionally inoculated in cooked cubed ham, packaged in Modified Atmosphere Packaging and stored at different temperatures. The bioprotective cultures limit L. monocytogenes growth in cubed cooked ham stored either at 4 °C for 60 days and at 4 °C for 20 days and at 8 °C for 40 days. The inhibition at 8 °C is particularly useful for industrial cooked meat products, considering there are often thermal abuse conditions (8 °C) in the supermarkets. Both the starters can eliminate L. monocytogenes risk and maintain the products safe, despite the thermal abuse conditions. In addition, both culture starters grew without producing perceptible sensory variations in the samples, as demonstrated by the panel of the untrained tasters. The bioprotective LAB produced neither off-odours and off-flavours, nor white/viscous patinas, slime, discoloration or browning. Therefore, according to the obtained data, and despite the fact that cooked cubed ham did not show pH ≤ 4.4 or aw ≤ 0.92, or pH ≤ 5.0 and aw ≤ 0.94, as cited in the EC Regulation 2073/2005. It can be scientifically stated that cubes of cooked ham with the addition of bioprotective starters cultures do not constitute a favourable substrate for L. monocytogenes growth. Consequently, these products can easily fall into category 1.3 (ready-to-eat foods that are not favourable to L. monocytogenes growth, other than those for infants and for special medical purposes), in which a maximum concentration of L. monocytogenes of 100 CFU g−1 is allowed.
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