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The aim of this study was to develop a novel and healthier fermented meat product by replacing pork fat with avocado pulp (AVP) during salami production. Experimental salamis were produced under laboratory conditions by substituting pork fat with AVP partially (10-AVP) and totally (20-AVP), while control salamis (CTR) remained AVP-free. The microbial composition of control and experimental salamis was assessed using a combined culture-dependent and -independent approach. Over a 20-days ripening period, lactic acid bacteria, coagulase-negative staphylococci, and yeasts dominated the microbial community, with approximate levels of 9.0, 7.0 and 6.0 log CFU/g, respectively. Illumina technology identified 26 taxonomic groups, with leuconostocs being the predominant group across all trials [constituting 31.26-59.12 % of relative abundance (RA)]. Gas Chromatography-Mass Spectrometry (GC-MS) analysis revealed changes in fatty acid composition and volatile organic compounds due to the substitution of pork fat with AVP. Specifically, monounsaturated fatty acids and terpene compounds increased, while saturated fatty acids and lipid oxidation products decreased. Although AVP influenced the sensory characteristics of the salamis, the highest overall satisfaction ratings were observed for the 10-AVP salamis. Consequently, substituting pork fat with AVP emerges as a viable strategy for producing healthier salamis and diversifying the meat product portfolio., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Typically, Swiss-type cheese is made from cow milk. However, in the present work an attempt to expand the sheep supply chain and product offering in this field was made by developing a new type of cheese using Swiss-type cheese technology. The cheese was manufactured under industrial conditions, and fermentations were carried out using freeze-dried commercial starters that are traditionally used in the production of Swiss cheese. Two experimental "Ewiss cheese" (EC) products were produced using raw milk (RM) and pasteurized milk (PM), respectively. Fourteen microbial groups were investigated by plate counts from curd until ripened cheeses. According to microbiological analyses, no statistically significant differences were found between the 2 productions with respect to the group of lactic acid bacteria (LAB). The curds were mainly characterized by mesophilic LAB cocci (7.45 log 10 cfu/g in RM-EC and 7.33 log 10 cfu/g in PM-EC). However, at the end of the ripening period (9 mo), the cheeses exhibited a higher presence of mesophilic LAB rods. Undesired microbiological groups were found only in the curd of raw milk cheese in the range of 10 4 to 10 5 cfu/g, but they were reaching undetectable levels by plate count in the cheese at the end of ripening. The RM-EC and PM-EC were characterized by 76% and 68% of DM, respectively. These cheeses contained 29.30% and 34.36% of protein, and 51.31% and 50.38% of fat, respectively. Textural analysis showed differences in terms of hardness, chewiness, and gumminess between the experimental cheeses and Swiss cheese sold on the market. These differences could be attributed to the higher protein content of ewe milk. The main fatty acids in the cheeses were palmitic acid, myristic acid, oleic acid, and capric acid. Among the organic acids, RM-EC had higher concentrations of lactic acid, whereas PM-EC was higher in propionic acid. The ewe cheeses emitted 46 volatile compounds, including acids, aldehydes, ketones, esters, alcohols, and other compounds. The PM-EC was characterized by the main compounds of Swiss-type cheese: acetic acid, butyric acid, ethyl butyrate, ethyl caproate, propanoic acid, and tetramethylpyrazine. Sensory evaluation showed that the new dairy products were generally appreciated, and PM-EC was the most preferred by the judges. This research has enabled the development of new ewe milk products, which could stimulate the valorization of a sector that has been long neglected and still has a large margin of improvement., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

In this study, 327 presumptive lactic acid bacteria (LAB) were isolated from goats' milk acid curds produced at a Sicilian dairy farm with the aim to identify potential starter cultures for traditional cheeses. All isolates were first processed by randomly amplified polymorphic DNA (RAPD)-PCR analysis. This approach identified 63 distinct strains which were evaluated for their acidifying capacity. Only 15 strains specifically stood out for their acidification capacity and were identified through 16S rRNA gene sequencing as Lactococcus lactis (11 strains) Enterococcus faecalis (three strains), and Ligilactobacillus animalis (one strain). Notably, all 15 LAB isolates produced bacteriocin-like inhibitory substances and anti-biofilm compounds, against both planktonic and biofilm forms of Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli, and Staphylococcus aureus, albeit at varying levels. Among these 15 LAB, En. faecalis RGM25 and Lc. lactis RGM55, susceptible to five antibiotics tested, were put in contact with wooden vat prototypes, because all equipment used in traditional cheese production in Sicily are made of wood. Scanning electron microscopy and bacterial plate counts of the wooden vat prototypes showed the development of biofilms at levels of approximately 6.0 log CFU/cm 2 . Overall, this study contributes to establishing a custom-made LAB starter cultures with bio-preservatives properties for Sicilian cheese productions., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The Castelvetrano method is the most widely used among the various table olive processing styles in Sicily. After debittering, the product is stored at low temperatures to prevent the growth of undesirable microorganisms. In an effort to enhance the production process, yeast isolates underwent genotypic characterization and technological screening. The screening process identified two yeast strains Candida norvegica OC10 and Candida boidinii LC1, which can grow at low temperatures and tolerate high pH values (up to 10) and salinity [10% (w/v)]. During the monitoring period, the inoculated trials showed limited presence of spoilage/pathogenic microorganisms. Additionally, the yeasts limited oxidative phenomena and softening of the drupes. The organic compounds detected were higher in the inoculated trials than in the control, and cold storage induced aromatic decay, which was less pronounced in the trial inoculated with C. norvegica. Sensory analysis revealed that the inoculated trials scored higher in sweetness, hardness and crispness., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Recently, the interest in improving livestock product nutraceutical profiles through sustainable feeding systems has increased. In this context, the overall quality and nutraceutical profiles were examined in dairy products obtained from 16 lactating Cinisara cows given an integrated feed in dry season with Opuntia ficus-indica cladodes. Two homogeneous groups of cows (milk yield: 6.3 ± 1.5 kg; body weight: 213 ± 55 kg) were fed with 2 different diets (CON: pasture and wheat bran; OFI: pasture, wheat bran, and cladodes), according to a 2 × 2 Latin square design. The bulk milk produced during the study was used to make Caciotta cheese and was analyzed at 0, 15, and 30 storage days. Milk and cheese samples were analyzed for chemical, physical, and microbiological traits. The nutraceutical and sensorial profiles, as well as the antioxidant capacity, were also determined in the final products. For milk, the urea content in individual samples was reduced in OFI but not in CON. In the cheese, integration of cladodes did not influence the starter cultures development with 2 strains of Streptococcus thermophilus, but it caused a higher content of polyphenols and a consequent greater antioxidant capacity and a change in the fatty acids profile. In particular, the caprylic, capric, lauric, myristic, and palmitic fatty acids were higher, as were the petroselinic, vaccenic, rumenic, and α-linolenic fatty acids. In contrast, the oleic and the γ-linolenic fatty acids were lower for OFI. The cheese from the OFI group showed better overall acceptability, and a higher yellow color, odor intensity, and butter flavor. The multivariate analysis well distinguished the cheeses belonging to the 2 groups. Further investigations should be conducted to formulate well-balanced diets that include cladodes for Cinisara lactating cows, but also to determine the content of other important bioactive compounds in fresh and treated cladodes, as well as their effects on animal welfare and production., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

This work was carried out with the aim to reduce the transformation duration of Protected Designation of Origin (PDO) Pecorino Siciliano cheese. To this purpose, the cooking in hot water (experimental production, EXP) was compared to the traditional cheese cooking under whey permeate (control production, CTR). The microbiological composition of under rind (UR) and core (Co) section of CTR and EXP cheeses was determined by a combined culture-dependent and -independent approach. Total mesophilic microorganisms and lactic acid bacteria (LAB) present in raw ewes' milk (5.0 log CFU/mL) increased during cheese making and reached values of about 8.0 log CFU/g in both sections (UR and Co) of 5-month ripened cheeses of both productions (CTR and EXP) monitored. The identification of the viable LAB populations in ripened cheeses showed that Enterococcus, Lacticaseibacillus, Lactiplantibacillus, Levilactobacillus, Limosilactobacillus and Streptococcus dominated UR and Co sections of all cheeses. MiSeq Illumina analysis demonstrated that LAB populations (lactobacilli, lactococci and streptococci) dominated the bacterial community of cheeses at 95.63-98.41 % of relative abundance. The two different cooking operations did not influence the physicochemical characteristics of PDO Pecorino Siciliano cheeses. Sensory evaluation performed by artificial senses analysis and trained panelists confirmed that the modification of PDO Pecorino Siciliano cheese production protocol did not significantly affect product characteristics and overall acceptance. Thus, data of this work confirmed that cooking under hot water allowed to reduce transformation duration and safeguard typicality of PDO Pecorino Siciliano cheese., Competing Interests: Declaration of competing interest No conflict of interest exists., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The rind acts as a protective barrier for internally-bacterial ripened cheeses. Unlike surface-inoculated smear cheeses, centripetal maturation is not assumed to occur in these cheeses. This research was aimed to evaluate the microbial diversity of the wooden shelves used for the ripening of Protected Denomination of Origin (PDO) Pecorino di Filiano and Protected Geographical Indication (PGI) Canestrato di Moliterno cheeses. The microorganisms associated with the rind of these cheeses were also investigated. Both wooden shelf surfaces and cheese rinds were sampled by brushing method to collect their biofilms. Wooden shelves showed levels of total mesophilic microorganisms (TMM) between 5.6 and 7.2 log CFU/cm 2 , while cheese rinds between 6.1 and 7.8 log CFU/cm 2 . The major dairy pathogens (Salmonella spp., Listeria monocytogenes, Escherichia coli, and Staphylococcus aureus) were never detected, while mesophilic and thermophilic bacteria dominated the surfaces of all wooden shelves and cheese rinds. LAB community was represented by Enterococcus spp., Leuconostoc spp., and Marinilactibacillus spp. Among yeasts, Debaryomyces spp., Candida spp., were identified, while Aspergillus spp., and Penicillium spp., dominated the community of filamentous fungi. MiSeq Illumina analysis identified 15 phyla, 13 classes, 28 orders, 54 families, and 56 genera among bacteria. Staphylococcus spp. was identified from all wooden surfaces, with a maximum abundance of 71 %. Brevibacterium, Corynebacterium and halophilic bacteria were detected in almost all samples. Regarding fungi, wooden shelves mainly hosted Aspergillus, Penicillium and Debaryomyces hansenii, while cheese rinds especially Penicillium and D. hansenii. Alpha diversity confirmed a strict correlation between the microbiota of wooden shelves and that of cheese rinds for the majority of factories. This study confirmed that the wooden shelves used for cheese ripening are microbiologically active and represent safe systems. Furthermore, the results of this work clarified the transfer flow between wooden shelves and PDO Pecorino di Filiano and PGI Canestrato di Moliterno cheese surfaces: smear-active microorganisms are mainly transferred from wooden shelves to cheese rind, which potentially contribute to the development of the final organoleptic characteristics; meanwhile, cheeses transfer LAB that are potentially involved in defining the safety aspects of the shelves., Competing Interests: Declaration of competing interest No conflict of interest exists., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

In recent years, a main goal of research has been to exploit waste from agribusiness industries as new sources of bioactive components, with a view to establishing a circular economy. Non-compliant avocado fruits, as well as avocado seeds and peels, are examples of promising raw materials due to their high nutritional yield and antioxidant profiles. This study aimed to recycle avocado food waste and by-products through dehydration to produce functional bread. For this purpose, dehydrated avocado was reduced to powder form, and bread was prepared with different percentages of the powder (5% and 10%) and compared with a control bread prepared with only semolina. The avocado pulp and by-products did not alter organoleptically after dehydration, and the milling did not affect the products' color and retained the avocado aroma. The firmness of the breads enriched with avocado powder increased due to the additional fat from the avocado, and alveolation decreased. The total phenolic content of the fortified breads was in the range of 2.408-2.656 mg GAE/g, and the antiradical activity was in the range of 35.75-38.235 mmol TEAC/100 g ( p < 0.0001), depending on the percentage of fortification.

This work was carried out with the aim to investigate the microbiological, physicochemical, and sensory properties of an innovative yoghurt produced from ewe's milk. Experimental yoghurt productions were performed with a commercial freeze-dried starter preparation and a natural milk starter culture (NMSC) of Streptococcus thermophilus and Lactobacillus delbrueckii . The two yoghurts did not differ for colour parameters, showing an average value of lightness, redness, and yellowness of 94.99, -3.74, and 9.37, respectively. The yoghurt produced using the NMSC as a fermenting agent was characterised by a significantly lower fat percentage and a higher antioxidant potential than commercial starters. Microbiological analysis confirmed the safety of the final product and a level of living lactic acid bacteria of 10 8 CFU/g. Sensory analysis revealed some differences among yoghurts regarding unpleasant odour, homogeneity, and persistence in the mouth, but the yoghurt processed with NMSC was more appreciated. Thus, the production of ewe's yoghurt fermented by a selected multi-strain starter culture represents an interesting strategy to enlarge the functional ovine dairy product portfolio.

Griffonia simplicifolia , a tropical plant endemic to West Africa, is highly regarded for its significant pharmacological potential. The objective of this study was to evaluate the metabolomic profile and to explore the antioxidant properties, antiproliferative activity, and antimicrobial potential of G. simplicifolia seed extracts obtained through either maceration, microwave-assisted extraction (MAE), or Soxhlet extraction using water, acetone, methanol and ethanol as solvents. Overall, methanol possessed superior total extraction efficiency. HPLC analyses confirmed the efficacy of acetone and ethanol as optimal solvents for the extraction of flavonoids and flavan-3-ols, whereas MAE exhibited enhanced effectiveness in extracting N-containing compounds, including 5-hydroxytryptophan (5-HTP). HPLC-MS analyses identified forty-three compounds, including thirty-four phenolic compounds and nine N-containing molecules. Isomyricitrin, taxifolin and a flavonol glucuronide were the main polyphenols, whereas 5-HTP was the main N-containing compound. Hydroalcoholic G. simplicifolia extracts showed the highest radical scavenging and metal-reducing antioxidant power, suggesting that most of the contribution to antioxidant activity depends on the more polar bioactive compounds. G. simplicifolia extracts showed dose-dependent antiproliferative activity against three distinct cancer cell lines (HeLa, HepG2, and MCF-7), with notable variations observed among both the different extracts and cell lines and divergent GI 50 values, emphasizing substantial discrepancies in cell sensitivity to the various extracts. Furthermore, G. simplicifolia extracts revealed antibiotic activity against Staphylococcus aureus . Our results highlight the potential of G. simplicifolia phytochemicals in the development of functional foods, nutraceuticals, and dietary supplements.

In this study, Caciocavallo Podolico Lucano (CPL) cheese was deeply characterized for its bacterial community, chemical composition and sensory aspects. The entire cheese making process (from milk collection to ripened cheese) was performed by strictly applying the traditional protocol for CPL production in four dairy factories (A-D) representative of the production area. The vat made of wood represents the main transformation tool for CPL cheese production and the biofilms hosted onto the internal surfaces of all vats analyzed in this study were dominated by lactic acid bacteria. Total mesophilic microorganisms present in bulk milk (4.7-5.0 log CFU/ml) increased consistently after contact with the wooden vat surfaces (5.4-6.4 log CFU/ml). The application of Illumina sequencing technology identified barely 18 taxonomic groups among processed samples; streptococci and lactobacilli constituted the major groups of the wooden vat biofilms [94.74-99.70 % of relative abundance (RA)], while lactobacilli dominated almost entirely (94.19-100 % of total RA) the bacterial community of ripened cheeses. Except coagulase positive staphylococci, undesirable bacteria were undetectable. Among chemical parameters, significant variations were registered for unsaturated, monounsaturated, polyunsaturated fatty acids and antioxidant properties (significantly lower for CPL cheeses produced in factory B). The cheeses from factories A, C and D were characterized by a higher lactic acid and persistence smell attributes than factory B. This work indicated that the strict application of CPL cheese making protocol harmonized the main microbiological, physicochemical and sensory parameters of the final cheeses produced in the four factories investigated., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

In the present work, oregano essential oils (OEOs) were applied to process the fresh ovine cheese "Tuma" obtained by pressed cheese technology. Cheese making trials were performed under industrial conditions using ewe's pasteurized milk and two strains of Lactococcus lactis (NT1 and NT4) as fermenting agents. Two experimental cheese products (ECP) were obtained through the addition of 100 (ECP100) and 200 (ECP200) µL/L of OEO to milk, while the control cheese product (CCP) was OEO-free. Both Lc. lactis strains showed in vitro and in vivo ability to grow in the presence of OEOs and to dominate over indigenous milk lactic acid bacteria (LAB) resistant to pasteurization. In the presence of OEOs, the most abundant compound found in cheese was carvacrol, constituting more than 65% of the volatile fraction in both experimental products. The addition of OEOs did not influence ash, fat, or protein content, but it increased by 43% the antioxidant capacity of the experimental cheeses. ECP100 cheeses showed the best appreciation scores by the sensory panel. In order to investigate the ability OEOs to be used as a natural preservative, a test of artificial contamination was carried out, and the results showed a significant reduction of the main dairy pathogens in OEO-added cheeses.

Almond production generates large amounts of by-products rich in polyphenols. In this study, almond skin was explored as a valuable food ingredient in bread making. To this purpose, almond skin was used to produce functional products modifying a traditional sourdough bread recipe. The doughs were prepared replacing semolina with powdered almond skin (PAS) at 5 and 10 % (w/w). Sourdough inoculum was started with a mix of lactic acid bacteria (LAB) and propagated in semolina until reaching pH 3.7. The pH of PAS added breads was higher than that of control (CTR) breads before and after fermentation. Plate counts showed a similar evolution of LAB and total mesophilic microorganisms, but members of Enterobacteriaceae and coliform were detectable in PAS doughs. Illumina data clearly showed a dominance of lactobacilli in all trials, but PAS doughs displayed the presence of Bacillus. The final bread characteristics were influenced by PAS and its addition percentage; in particular, crust and crumb colour resulted darker, the alveolation decreased and, regarding sensory attributes, odour intensity increased, while bread odour diminished. In presence of PAS, bread emissions were characterized by lower percentages of alcohols and aromatic hydrocarbons and higher percentages of the other volatile compound classes, especially terpenoids like β-pinene, β-myrcene and limonene than CTR trial. After in vitro simulated digestion, the final release of phytochemicals from 10 % PAS bread was almost 100 %. Thus, PAS determined an increase of the antioxidant capacity of the breads. Phytochemicals released from digested PAS-fortified bread can provide antioxidant protection in a complex biological environment such as human intestinal-like cells. Besides the positive functional properties of PAS, this work also evidenced the hygienic issues of almond skin and, in order to avoid potential risks for the human health, highlighted the need to preserve its microbiological characteristics during storage for their reuse in bread production., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

In this study, PDO Provola dei Nebrodi cheese was deeply characterized for its bacterial community and chemical composition. Four dairy factories (A-D) were monitored from milk to ripened cheese. Wooden vat biofilms were dominated by thermophilic rod LAB (4.6-6.5 log CFU/cm 2 ). Bulk milk showed consistent levels of total mesophilic microorganisms (TMM) (5.0-6.0 log CFU/mL) and, after curdling, a general increase was recorded. The identification of the dominant LAB in wooden vat biofilms and ripened cheeses showed that the majority of wooden vat LAB were lactococci and Streptococcus thermophilus, while cheese LAB mainly belonged to Lacticaseibacillus paracasei and Enterococcus. Illumina sequencing identified 22 taxonomic groups; streptococci, lactococci, lactobacilli and other LAB constituted the majority of the total relative abundance % of the wooden vat (69.01-97.58 %) and cheese (81.57-99.87 %) bacterial communities. Regarding chemical composition, the effect of dairy factories was significant only for protein content. Inside cheese color was lighter and yellower than surface. Differences in fatty acids regarded only myristic acid and total amount of monounsaturated fatty acids. The sensory evaluation indicated some differences among cheeses produced in the four dairies regarding color, homogeneity of structure, overall intensity, salty, spicy, and hardness. The integrated approach applied in this study showed that PDO Provola dei Nebrodi cheese characteristics are quite stable among the dairy factories analyzed and this has to be unavoidably imputed to the application of the same cheese making protocol among different dairies., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In this study, almond skin of Prunus dulcis L. variety Casteltermini was characterised for its chemical composition and for the inhibitory activity towards food associated microorganisms belonging to pathogenic ( Salmonella Enteritidis, Staphylococcus aureus , Escherichia coli and Listeria monocytgenes ), spoilage ( Pseudomonas poae and Brochotrix thermospacta ) and useful ( Levilactobacillus brevis and Saccharomyces cerevisiae ) groups. The n -hexane extract of P. dulcis skin, investigated by GC-MS, evidenced linoleic, palmitic, and oleic acids as the main abundant compounds. Staphylococcus aureus ATCC 33862 growth was strongly limited by n -hexane extract and the minimum inhibition concentration (MIC) was 25.5 mg/mL. These results highlight the potential of almond by-products as food bio-preservatives as well as pharmaceutical products.

The aim of this study was to evaluate the fermentation performance of the commercial starter Lactiplantibacillus pentosus OM13 with four nutrients (A, B, C, and D) that differed in the following ingredients: starch, sugars, maltodextrin, inactivated yeast, inactivated yeast rich in amino acids, inactivated yeast rich in mannoproteins, and salt (NaCl). For this purpose, six different experimental productions of Nocellara del Belice table olives were carried out. During transformation, the fermentation process was monitored by measuring pH and plate counts for lactic acid bacteria (LAB), yeasts, Enterobacteriaceae, Staphylococcaceae, and Pseudodomondaceae populations. At the end of the production process, each trial was subjected to volatile organic compound analysis and sensory evaluation. The addition of the different nutrients resulted in a significant reduction in pH (around 2.5 points) after 3 days of fermentation. At the same time, a significant increase in the number of LAB populations (> 6.6 log CFU/mL) was observed for all trials. Volatile organic compound (VOC) analysis revealed the presence of 39 compounds. In this study, nutrient C was optimal for improving the fermentation activity of L. pentosus OM13. These results provide elements for the implementation of experimental protocols to reduce product losses and improve sensory characteristics., Competing Interests: The authors declare no conflict of interest.

Catarratto is one of the most widely cultivated grape varieties in Sicily. It is an indigenous non-aromatic white grape variety. Despite its widespread use in winemaking, knowledge of the aroma and chemical and microbiological properties of Catarratto wines is quite limited. The influence of Metschnikowia pulcherrima combined with Saccharomyces cerevisiae on the aromatic expression of Catarratto wines was investigated with and without the addition of glutathione-rich inactivated yeast. The substance is a natural specific inactivated yeast with a guaranteed glutathione level used to limit oxidative processes. The aromatic profiles of the final wines were determined through analysis of the volatile organic compounds using a solid-phase microextraction technique that identified 26 aromatic compounds. The addition of M. pulcherrima in combination with the natural antioxidant undoubtedly increased the aromatic complexity of the wines. Dodecanal was exclusively detected in the wines processed with glutathione-rich inactivated yeasts. Furthermore, the presence of this natural antioxidant increased the concentration of six esters above the perception threshold. Sensory analysis was also performed with a panel of trained judges who confirmed the aromatic differences among the wines. These results suggest the suitability of glutathione-rich inactivated yeasts for determining the oxidative stability of Catarratto wines, thus preserving its aromatic compounds and colour.

Bacteria producers of plant growth-promoting (PGP) substances are responsible for the enhancement of plant development through several mechanisms. The purpose of the present work was to evaluate the PGP traits of 63 bacterial strains that were isolated from an anthropogenic soil, and obtained by modification of vertisols in the Sicily region (Italy) seven years after creation. The microorganisms were tested for the following PGP characteristics: indole acetic acid (IAA), NH 3 , HCN and siderophore production, 1-aminocyclopropane-1-carboxylate deaminase activity (ACC) and phosphate solubilization. The results of principal component analysis (PCA) showed that Bacillus tequilensis SI 319, Brevibacterium frigoritolerans SI 433, Pseudomonas lini SI 287 and Pseudomonas frederiksbergensis SI 307 expressed high levels of IAA and production of ACC deaminase enzyme, while for the rest of traits analyzed the best performances were registered with Pseudomonas genus, in particular for the strains Pseudomonas atacamensis SI 443, Pseudomonas reinekei SI 441 and Pseudomonas granadensis SI 422 and SI 450. The in vitro screening provided enough evidence for future in vivo growth promotion tests of these eight strains.

The demand for unique and exclusive food products and beverages is constantly on the increase. One of the products that mostly evolved to encounter market dynamics in the last decade is craft beer. For a long time, craft breweries have included fruit in beer production to enrich flavour and aroma profile of different beer styles. In this study, for the first time, the use of Saccharomyces and non-Saccharomyces yeast strains isolated from high-sugar matrices (manna and fermented honey by-products) were investigated to diversify fruit craft beer production, in order to improve the fermentation process and highlight the complexity of aroma profiles generated during alcoholic fermentation. Two yeast strains, Hanseniaspora uvarum YGA34 and Saccharomyces cerevisiae MN113, were tested as co-starters and starters for their beer production capacity. Commercial yeast strain US-05 was used as control. Loquat juice was added at the end of primary alcoholic fermentation in all trials. Interestingly, S. cerevisiae MN113 consumed sugars faster than control strain S. cerevisiae US-05, including maltose, even in the case of sequential inoculation. This strain showed an excellent ability to consume rapidly sugars present. All strains showed their concentrations ranged between 5 and 8 Log cycles during fermentation. The absence of off-odours and the improvement of aromatic perception were observed in experimental trials involving the use of S. cerevisiae MN113 as a monoculture and in sequential combination with H. uvarum YGA34. Esters and alcohols were the most abundant compounds emitted from the beers. The beers produced with sequential inoculation of H. uvarum YGA34 and S. cerevisiae MN113 or US-05 are characterised by a higher ester and lower alcohol concentration. These two unconventional yeast strains from high sugar matrices showed great technological properties, representing promising co-starters and starter during craft fruit beer production., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The aim of this study was to produce a fresh ovine pressed cheese within Pecorino "Primosale" typology with the addition of citrus essential oils (EOs). For this purpose, ewe's pasteurized milk was added with EOs from the peel of lemons, oranges and tangerines. Seven cheese productions were performed at the pilot plant scale level, including one control production (CP) without the addition of EOs and six experimental productions obtained by the addition of two EO concentrations (100 and 200 µL/L) to milk. The acidification process was obtained by means of the starter cultures Lactococcus lactis CAG4 and PON36. All cheeses showed levels of lactic acid bacteria (LAB) around 10 9 CFU/g, indicating that citrus EOs did not negatively influence the starter evolution. The addition of citrus EOs did not determine significant variations for dry matter, fat and protein percentages but increased the antioxidant capacity of all the experimental cheeses of about 50% in comparison to the control trial. The citrus EOs impacted cheese VOCs, especially for terpene class (limonene, β-pinene, myrcene, carene, linalool and α-terpineol). The sensory evaluation showed that cheeses enriched with 100 µL/L of citrus EOs were mostly appreciated by the panelists.

Mead is a beverage produced by alcoholic fermentation of honey-must. The starter yeasts that are commonly used for the alcoholic fermentation of honey-must are oenological Saccharomyces cerevisiae strains. The objective of the present work was, for the first time, to apply yeasts of honey by-products origin to evaluate the influences the taste-olfactory attributes of mead. For this purpose, three experimental productions were set up, which included: (i) single inoculation of S. cerevisiae; (ii) single inoculation of Hanseniaspora uvarum; (iii) sequential inoculation of H. uvarum/S. cerevisiae. Two control trials were performed, using a commercial strain of S. cerevisiae of oenological origin and a spontaneous fermentation. The results of the chemical parameters showed differences between the trials in terms of residual sugars, acetic acid, glycerol, ethanol and volatile organic compounds. Sensorial analysis also showed a high heterogeneity among trials. The attributes of sweetness, honey and floral were found in mead fermented with H. uvarum, whereas all meads obtained with S. cerevisiae were dry, balanced and without off-odors and off-flavours. The results obtained showed that the controlled application of conventional and non-conventional yeast strains isolated from honey by-products origin could be a promising approach to improve the quality of meads., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In the context of the contemporary research on sustainable development and circular economy, the quest for effective strategies aimed at revaluation of waste and by-products generated in industrial and agricultural production becomes important. In this work, an ethanolic extract from red raspberry ( Rubus idaeus ) seed waste (WRSP) was evaluated for its phytochemical composition and functional properties in term of antioxidative, antiproliferative, and antimicrobial activities. Chemical composition of the extract was determined by both HPLC-ESI-MS/MS and spectrophotometric methods. Phytochemical analysis revealed that flavan-3-ols and flavonols were the major phenolic compounds contained in WRSP. The extract demonstrated very high radical-scavenging (4.86 ± 0.06 µmol TE/DW) and antioxidant activity in a cell-based model (0.178 ± 0.03 mg DW/mL cell medium). The WRSP extract also exhibited antiproliferative activity against three different epithelial cancer cell lines (MCF-7, HepG2, and HeLa cells) in a dose-dependent manner. Finally, microbiological assays showed the absence of colonies of bacteria and microscopic fungi (yeasts and molds) and revealed that the WRSP extract has a large inhibition spectrum against spoilage and pathogenic bacteria, without inhibitory activity against pro-technological bacteria. In conclusion, the obtained results show that WRSP is a rich source of phytochemical compounds exerting interesting biological activities. For these reasons WRSP could find applications in the nutritional, nutraceutical, and pharmacological fields.

Aims: The purpose of this study was to functionalize an ovine stretched cheese belonging to 'Vastedda' typology with red grape pomace powder (GPP) of Nero d'Avola cultivar and to characterize the microbiological, physicochemical, phenolic profile and sensory characteristics of the final cheeses., Methods and Results: Before cheeses production, GPP was characterized for its microbiological profile, antibacterial activity and polyphenolic content. No colonies of bacteria and yeasts were detected in the GPP. GPP showed a large inhibition spectrum against spoilage and pathogenic bacteria. Three classes of polyphenolic compounds belonging to flavan-3-ols, flavonol and phenolic acids were identified. Two cheeses [0 and 1% (w w -1 ) of GPP] were produced with pasteurized ewe's milk and commercial starter cultures. Plate counts and randomly amplified polymorphic DNA analysis demonstrated the ability of the starter strains to drive the fermentation process in the presence of GPP. GPP enrichment resulted in an increase of protein, phenolic compounds, sensory traits and reduced fat., Conclusions: GPP addition to cheese represents an optimal strategy for the valorization of winemaking by-products and to obtain polyphenol-enriched cheese., Significance and Impact of the Study: This study allowed to achieve an ovine cheese with specific physicochemical, nutraceutical and sensorial characteristics able to enlarge the functional dairy product portfolio., (© 2021 The Society for Applied Microbiology.)

This research aimed to analyze variations in chemical properties, microbiological characteristics and generated volatile organic compounds (VOCs) profile during sourdough fermentation. Sourdoughs were collected from different cities in Turkey at two different times and lactic acid bacteria (LAB) in the samples were identified with culture-independent and culture-dependent molecular methods. According to culture-dependent methodology, thirteen LAB species were identified. Lactobacillus spp. were identified as the major group according to MiSeq Illumina analysis. Technological potential of commonly isolated LAB species was evaluated. Due to high frequency of isolation, Fructilactobacillus sanfranciscensis and Lactiplantibacillus plantarum strains were better investigated for their technological traits useful in sourdough production. Experimental sourdoughs were produced with mono- and dual-culture of the selected strains and chemical properties and microbiological characteristics, as well as VOCs profile of the sourdoughs, were subjected to multivariate analysis which showed the relevance of added starter, in terms of acidification and VOCs profile., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

"Spiritu re fascitrari" is a Sicilian alcoholic beverage obtained through distillation of a decoction of spontaneously fermented honey by-products (FHP). The production process often leads to sensorial defects due to the unstable alcoholic fermentation. The objective of this work was to select Saccharomyces cerevisiae strains from FHP to be used as starter in decoction fermentation. Based on chemical, microbiological and technological data, from a total of 91 strains three S. cerevisae were selected for further testing to produce FHP at laboratory scale level. After FHP distillation, the analysis of volatile organic compounds showed a complex mixture of sensory active molecules, mainly alcohols and aldehydes. Among the alcohols, 3-methyl-1-butanol, 2-methyl-1-butanol, phenylethyl alcohol, hexadecanol and octadecanol were found at the highest concentrations. Among the carboxylic acids, acetic acid was mainly detected in the spontaneously fermented samples. FHP fermented with the three selected strains were not characterized by the presence of off-odors or off-flavours. The results obtained in this work demonstrate that the selected S. cerevisiae strains are promising starters to stabilize the production of distilled alcoholic beverages produced from honey by-products., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

During a survey for isolating sourdough lactic acid bacteria (LAB), 20 dough samples produced at the bakery level (BL) or home-made (HM) were collected. An enzyme-based colorimetric method revealed a total biogenic amines (BAs) concentration in the range 41.4-251.8 ppm for six (three BL and three HM) sourdoughs characterised by unpleasant odours. Eight BAs generally investigated in foods were identified and quantified from these six samples by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Only one HM sample contained almost all analysed BAs. Tryptamine was exclusively detected in HM sourdoughs (0.71-24.1 ppm). Putrescine, tryptamine, spermidine, and spermine were the only BAs detected in BL sourdoughs. MiSeq Illumina analysis was applied to study the total bacterial community of sourdoughs. LAB accounted from 67.89 to 92.17% of total bacterial diversity, and Levilactobacillus brevis was identified in all six sourdoughs. Leuconostoc , Pediococcus , and Weissella were also dominant. Plate counts detected neither the presence of Pseudomonas nor members of the Enterobacteriaceae family, and LAB levels were, on average, barely 5.89 Log CFU/g for BL, and 7.33 Log CFU/g for HM sourdoughs. Data suggested that the microorganisms mainly imputable of BAs formation in sourdough are members of the LAB community.

The present study was carried to correlate the microbial diversity of the biofilms developed on the wooden boards used for aging traditional Sicilian cheeses with cheese typology. To this end, the microbial diversity of the shelves in contact with the cheeses PDO Pecorino Siciliano, PDO Piacentinu Ennese, and TAP Caciocavallo Palermitano, during ripening, was evaluated by a multivariate statistical approach. The shelf biofilms of this study were previously analyzed for their microbial composition, but no correlation between biodiversity and cheese type was investigated. Canonical discriminant analysis confirmed a cheese typology effect on the microbial loads of the wooden shelves investigated. Regarding the plate count data, the centroids of different cheeses were statistically distant from one another. This analysis also showed a good graphic separation of data regarding bacterial order operational taxonomy units (OTUs). Thus, the microbiological differences imputed to the cheese typologies were not affected by the environmental conditions of the facilities. Furthermore, wooden shelf lactic acid bacteria (LAB) were investigated for their ability to inhibit the main dairy pathogens. Although inhibitors were mainly enterococci, P. pentosaceus WS287 and W. paramesenteroides WS581 showed the highest inhibition activity, indicating their possible application to control the undesired bacteria in situ.

This work was performed to produce a new soft ewe's milk cheese, namely "Quadrello di ovino" (QdO) cheese, to enlarge ewe's dairy product portfolio of South Italy, barely limited to Pecorino cheese typology. Cheese making was performed applying the technology for "Crescenza" cheese typology with some modifications. In particular, pasteurized ewes' milk was inoculated with two commercial starter formulations (SF1 and SF2) of Streptococcus thermophilus to obtain two different productions (QdO-P1 and QdO-P2, respectively). Plate counts demonstrated the ability of both starter formulations to drive the fermentation process, since S. thermophilus counts reached 10 9 CFU/g in both productions. Generally, the two starter formulations did not affect the chemical composition of QdO cheeses that contained, on average, 64.08% dry matter of which approximately 54.99% were fats and 36.39% proteins. Among chemical parameters, significant differences were registered for secondary lipid oxidation state (significantly lower for QdO-P2), fatty acids and volatile organic compounds (VOCs). However, the differences registered among cheese VOCs from were not perceived by the panelists who recognized both cheese productions highly similar, although QdO-P2 cheeses were mostly appreciated by the judges. This study allowed to produce a novel fresh ovine cheese with specific chemical and sensorial characteristics well appreciated by consumers.

Catarratto is one of the most common non-aromatic white grape varieties cultivated in Sicily (Southern Italy). In order to improve the aromatic expression of Catarratto wines a trial was undertaken to investigate the effect of yeast strain, nutrition and reduced glutathione. Variables included two Saccharomyces cerevisiae strains, an oenological strain (GR1) and one isolated from honey by-products (SPF52), three different nutrition regimes (Stimula Sauvignon Blanc™ (SS), Stimula Chardonnay™ (SC) and classic nutrition practice), and a specific inactivated yeast rich in reduced glutathione to prevent oxidative processes [Glutastar™ (GIY)] ensuing in ten treatments (T1-T10). Microbiological and chemical parameters demonstrated the aptitude of strain SPF52 to successfully conduct alcoholic fermentation. During fermentation, the Saccharomyces yeast populations ranged from 7 to 8 logarithmic CFU/mL. All wines had a final ethanol content ranging between 12.91 and 13.85% (v/v). The dominance of the two starter strains over native yeast populations was higher than 97% as estimated by interdelta analysis. The addition of nutrients SS or SC increased the aromatic complexity of the wines as reflected by volatile organic compounds (VOCs) composition and sensory profiles. In particular, 32 VOCs were identified; alcohols (62.46-81.1%), thiols (0.27-0.87%), ethers (0.09-0.16%), aldehydes (0-1.21%), ketones (0-2.28%), carboxylic acids (4.21-12.32%), esters (0-10.85%), lactones (0.9-1.49%) and other compounds (0.77-6.9%). Sensory analysis demonstrated a significant impact on wine aroma in relation to yeast starter strain used, the type of nutrition (SS, SC or classic nutrition) and the presence/absence of GIY. The wines produced with GR1 yeast strain and SS (T2), SPF52 with SC (T9) both in presence of GIY showed higher overall quality. Trials T2 and T9 showed the highest scores for 13 and 18 attributes, respectively. The different nutrition, addition of GIY and the yeast starter strains diversified and enhanced sensory expression of Catarratto wines., (Copyright © 2021 Elsevier B.V. All rights reserved.)

In this work, the feasibility of antibacterial biopolymeric films containing carvacrol (CRV) and a nisin commercial formulation (Nis) for potential food packaging applications was investigated. As polymer matrix, a commercial biodegradable polymer formulation of Mater-Bi (MB) was chosen due to its significant food packaging applications. CRV and Nis were chosen due to their well-established antibacterial properties and their potential synergistic effect. MB/CRV, MB/Nis, and MB/CRV/Nis systems were produced by melt mixing and compression molding. The mechanical properties of the films were evaluated by tensile tests. Differential scanning calorimetry was assessed aiming at investigating the effect of the two compounds and their mixture on the thermal properties of MB. The release profile of CRV and Nis from the MB-based films was evaluated in water at 4 °C by UV-Vis measurements and it was fitted with a power-law model. The antibacterial activity of MB-based films was tested in vitro against Listeria monocytogenes, Salmonella enteritidis, Escherichia coli, and Staphylococcus aureus. The combination of CRV and Nis strongly affected the properties of the MB-based films and ensured higher antibacterial activity if compared to MB/CRV and MB/Nis systems., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The main goal of this research was to characterize the bacterial diversity of the wooden boards used for aging traditional Sicilian cheeses and to evaluate whether pathogenic bacteria are associated with these surfaces. Eighteen cheese dairy factories producing three traditional cheese typologies (PDO Pecorino Siciliano, PDO Piacentinu Ennese, and Caciocavallo Palermitano) were selected within the region of Sicily. The wooden shelf surfaces were sampled by a destructive method to detach wood splinters as well as by a nondestructive brushing to collect microbial cells. Scanning electron microscopy showed the presence of almost continuous bacterial formations on the majority of the shelves analyzed. Yeasts and fungal hyphae were also visualized, indicating the complexity of the plank communities. The amplicon library of the 16S rRNA gene V3-V4 region was paired-end sequenced using the Illumina MiSeq system, allowing the identification of 14 phyla, 32 classes, 52 orders, 93 families, and 137 genera. Staphylococcus equorum was identified from all wooden surfaces, with a maximum abundance of 64.75%. Among cheese-surface-ripening bacteria, Brevibacterium and Corynebacterium were detected in almost all samples. Several halophilic ( Halomonas , Tetragenococcus halophilus, Chromohalobacter, Salimicrobium , Marinococcus , Salegentibacter , Haererehalobacter , Marinobacter , and Idiomarinaceae ) and moderately halophilic ( Salinicoccus , Psychrobacter , and Salinisphaera ) bacteria were frequently identified. Lactic acid bacteria (LAB) were present at low percentages in the genera Leuconostoc , Lactococcus , Lactobacillus , Pediococcus , and Streptococcus. The levels of viable microorganisms on the wooden shelves ranged between 2.4 and 7.8 log CFU/cm 2 . In some cases, LAB were counted at very high levels (8.2 log CFU/cm 2 ). Members of the Enterobacteriaceae family were detected in a viable state for only six samples. Coagulase-positive staphylococci, Salmonella spp., and Listeria monocytogenes were not detected. Seventy-five strains belonged to the genera Leuconostoc , Lactococcus , Pediococcus , Enterococcus , Lactobacillus , and Weissella . IMPORTANCE This study provides evidence for the lack of pathogenic bacteria on the wooden shelves used to ripen internal bacterially ripened semihard and hard cheeses produced in Sicily. These three cheeses are not inoculated on their surfaces, and surface ripening is not considered to occur or, at least, does not occur at the same extent as surface-inoculated smear cheeses. Several bacterial groups identified from the wooden shelves are typically associated with smear cheeses, strongly suggesting that PDO Pecorino Siciliano, PDO Piacentinu Ennese, and Caciocavallo Palermitano cheese rind contributes to their final organoleptic profiles.

The increasing interest in novel beer productions focused on non-Saccharomyces yeasts in order to pursue their potential in generating groundbreaking sensory profiles. Traditional fermented beverages represent an important source of yeast strains which could express interesting features during brewing. A total of 404 yeasts were isolated from fermented honey by-products and identified as Saccharomyces cerevisiae, Wickerhamomyces anomalus, Zygosaccharomyces bailii, Zygosaccharomyces rouxii and Hanseniaspora uvarum. Five H. uvarum strains were screened for their brewing capability. Interestingly, Hanseniaspora uvarum strains showed growth in presence of ethanol and hop and a more rapid growth than the control strain S. cerevisiae US-05. Even though all strains showed a very low fermentation power, their concentrations ranged between 7 and 8 Log cycles during fermentation. The statistical analyses showed significant differences among the strains and underlined the ability of YGA2 and YGA34 to grow rapidly in presence of ethanol and hop. The strain YGA34 showed the best technological properties and was selected for beer production. Its presence in mixed- and sequential-culture fermentations with US-05 did not influence attenuation and ethanol concentration but had a significant impact on glycerol and acetic acid concentrations, with a higher sensory complexity and intensity, representing promising co-starters during craft beer production., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This work was carried out to elaborate selenium (Se) bio-enriched fermented Mediterranean fruit juices. To this purpose, pomegranate and table red grape juices were added with sodium selenite (Na 2 SeO 3 ) and fermented by Levilactobacillus brevis CRL 2051 and Fructobacillus tropaeoli CRL 2034 individually or combined. To better evaluate the effect of selenite addition and starter strain inoculums on the total bacterial community of the fruit juices, fermentation trials were performed with raw and pasteurized fruit juices. No statistical significant differences were observed for total mesophilic microorganisms (TMM) and rod-shaped lactic acid bacteria (LAB) levels among raw and pasteurized juices inoculated with the starter strains, while significant differences between those juices with and without selenite were registered. LAB cocci, Pseudomonadaceae and yeasts were detected only for the raw juice preparations. The dominance of L. brevis CRL 2051 and F. tropaeoli CRL 2034 was confirmed by randomly amplified polymorphic DNA (RAPD)-PCR analysis. After fermentation, pH dropped for all inoculated trials and control raw juices. The soluble solid content (SSC) levels of the raw juices were higher than the corresponding pasteurized trials. The thermal treatment affected consistently yellowness of grape juice trials and redness of pomegranate juices. No microbial Se accumulation was registered for pomegranate juices, while F. tropaeoli CRL 2034 accumulated the highest amount of Se (65.5 μg/L) in the grape juice. For this reason, only trials carried out with raw grape juices were investigated by metagenomics analysis by Illumina MiSeq technology. Non-inoculated grape juices were massively fermented by acetic acid bacteria while Fructobacillus and Lactobacillus (previous genus name of Levilactobacillus) represented the highest operational taxonomy units (OTUs) relative abundance % of the trials inoculated with the starter strains as confirmed by this technique., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Fresh ovine "primosale" cheese was processed with the addition of grape pomace powder (GPP). Cheese making was performed using pasteurized ewes' milk and four selected Lactococcus lactis strains (Mise36, Mise94, Mise169 and Mise190) inoculated individually. For each strain the control cheese (CCP) was not added with GPP, while the experimental cheese (ECP) was enriched with 1% ( w / w ) GPP. GPP did not influence the starter development that reached levels of 10 9 CFU/g in all final cheeses. The comparison of the bacterial isolates by randomly amplified polymorphic DNA (RAPD)-PCR showed the dominance of the added strains over indigenous milk bacteria resistant to pasteurization. GPP addition reduced fat content and determined an increase of protein and of secondary lipid oxidation. Sensory tests indicated that cheeses CCP94 and ECP94, produced with the strain Mise94, reached the best appreciation scores. Following in vitro simulated human digestion, bioaccessible fraction of ECP94 showed antioxidant capacity, evaluated as radical scavenging activity and inhibition of membrane lipid oxidation, significantly higher than that from CCP94, with promising increase in functional properties. Thus, the main hypothesis was accepted since the functional aspects of the final cheeses improved, confirming that GPP is relevant for sustainable nutrition by using winemaking by-products.

An innovative ovine cheese enriched with red grape pomace powder (GPP) was produced to improve the functional properties of Vastedda cheese typology. Vastedda cheese making was performed adding GPP and four selected Lactococcus lactis strains (Mise36, Mise94, Mise169 and Mise190). For each strain, 40 L of pasteurized ewe's milk was divided into two aliquots representing control and experimental trials. Control cheese (CC) production did not contain GPP, while the experimental cheese (EC) production was enriched with 1% ( w / w ) GPP. GPP did not slow down starter development and acid generation. Plate counts and randomly amplified polymorphic DNA (RAPD)-PCR analysis confirmed the dominance of the starters in all trials. The evolution of the physicochemical parameters showed that EC productions were characterized by lower fat content, higher protein content, and higher values of secondary lipid oxidation. Sensory evaluation indicated that the cheeses produced with the strain Mise94 were those more appreciated by the judges. Thus, the last cheeses were investigated for some functional aspects: GPP enrichment significantly increased antioxidant activity and lipoperoxyl radical scavenger capacity, confirming that grape polyphenol inclusion in cheese represents an optimal strategy for the valorization of ovine cheeses as well as winemaking industry by-products.

This review article focuses on the technological aspects and microbiological critical points of pressed-cooked cheeses processed from raw ewe's milk without the inoculation of starter cultures, in particular "Pecorino" cheese typology produced in Italy. After showing the composition of the biofilms adhering to the surface of the traditional dairy equipment (mainly wooden vat used to collect milk) and the microbiological characteristics of PDO Pecorino Siciliano cheese manufactured throughout Sicily, this cheese is taken as a case study to develop a strategy to improve its hygienic and safety characteristics. Basically, the natural lactic acid bacterial populations of fresh and ripened cheeses were characterized to select an autochthonous starter and non-starter cultures to stabilize the microbial community of PDO Pecorino Siciliano cheese. These bacteria were applied at a small scale level to prove their in situ efficacy, and finally introduced within the consortium for protection and promotion of this cheese to disseminate their performances to all dairy factories. The innovation in PDO Pecorino Siciliano cheese production was proven to be respectful of the traditional protocol, the final cheeses preserved their typicality, and the general cheese safety was improved. An overview of the future research prospects is also reported.

The present work was carried out to investigate the Antimicrobial Resistance (AMR) of enterococci isolated from raw ewes' and cows' milk. The samples were collected from eighteen semi-extensive dairy sheep and cow farms throughout western Sicily. Plate counts, carried out on Rapid Enterococcus Agar commonly used to detect food enterococci, revealed a maximal enterococcal concentration of approximately 4.58 Log Colony Forming Unit (CFU)/mL. Colonies were isolated and differentiated based on genetic analysis by Randomly Amplified Polymorphic DNA (RAPD)-PCR. Thirty-eight different strains were identified. Analysis by a species-specific multiplex PCR assay grouped the strains into three Enterococcus species such as Enterococcus durans , Enterococcus faecalis and Enterococcus faecium . The 38 strains were also investigated for their antimicrobial resistance by a phenotypic approach. All 38 Enterococcus displayed resistance to at least one or more of the antimicrobials tested confirmed that the dairy enterococci could be a vector for the dissemination of antimicrobial resistance. This work showed that enterococci with AMR traits are commonly present in semiextensive dairy sheep and cow farms of western Sicily pointed out the relevance of informing dairy makers and veterinary regarding the antimicrobial use in order to mitigate problems of public health and veterinary medicine., Competing Interests: Conflict of interests: The authors declare no conflict of interest., (©Copyright: the Author(s).)

Fermented chickpea liquid is used as a leavening agent in chickpea bread production. In the present study, traditional chickpea liquid starter and dough samples were collected from bakeries in Turkey and microbiologically investigated. Culture-independent analysis for microbiota diversity, performed by MiSeq Illumina, identified Clostridium perfringens as major group in all samples, while Weissella spp. Dominated LAB community. A culture-dependent methodology was applied and 141 isolates were confirmed to be members of the LAB group based on 16s rRNA gene sequence analysis. In particular, 11 different LAB species were identified confirming the high frequency of isolation of weissellas, since Weissella confusa and Weissella cibaria constituted 47.8 and 12.4%, respectively, of total LAB isolated. The other species were Enterococcus faecium, Enterococcus lactis, Lactobacillus brevis, Lactobacillus plantarum, Leuconostoc mesenteroides, Leuconostoc mesenteroides subsp. Dextranium, Pediococcus acidilactici, Pediococcus pentosaceus and Streptococcus lutetiensis. Due to high frequency of isolation, W. confusa strains were investigated at technological level and W. confusa RL1139 was used as mono-culture starter in the experimental chickpea sourdough production. Chemical and microbiological properties, as well as volatile organic compounds (VOCs) of the chickpea liquid starters and doughs were subjected to a multivariate analysis. Control and W. confusa inoculated chickpea liquid starter and dough samples were close to each other in terms of some characteristics related to chemical, microbiological and VOCs profile, but the inoculated sourdough showed a higher generation of certain VOCs, like butanoic acid (81.52%) and ethyl acetate (8.15%) than control sourdough. This is important in order to maintain typical characteristics of the traditional chickpea dough, but at the same time improving the aroma profile. This work demonstrated that W. confusa RL1139 can be applied at large scale production level without compromising the typical characteristics of the final product., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The present study was carried out to produce Protected Denomination of Origin (PDO) Pecorino Siciliano cheese with a multi-species lactic acid bacteria (LAB) culture, composed of starter and non-starter strains in order to reduce the microbiological variability of the products derived without LAB inoculums. To this end, cheese samples produced in six factories located in five provinces (Agrigento, Catania, Enna, Palermo and Trapani) of Sicily, and previously characterised for physicochemical, microbiological and sensory aspects, have been investigated in this work for bacterial microbiome, fatty acid (FA) composition as well as volatile organic compound (VOC) profiles. Analysis of the cheese microbiomes indicated that streptococci (30.62-77.18% relative abundance) and lactobacilli (on average 25.90% relative abundance) dominated the bacterial communities of control cheeses, produced without exogenous inoculums, whereas the cheeses produced with the selected multi-strain culture saw the dominance of lactococci (in the range 6.49-14.92% relative abundance), streptococci and lactobacilli. After the addition of the selected mixed culture, Shannon index increased in all cheeses, but only the cheeses produced with the selected LAB mixed culture in the factory 2 showed Gini-Simpson diversity index (0.79) closer to the reference value (0.94) for a perfect even community. FA composition, mainly represented by saturated FA (on average 69.60% and 69.39% in control cheeses and experimental cheeses, respectively), was not affected by adding LAB culture. The presence of polyunsaturated FA ranged between 7.93 and 8.03% of FA. VOC profiles were different only for the content of butanoic acid, registered for the experimental cheeses at higher concentrations (on average 662.54 mg/kg) than control cheeses (barely 11.96 mg/kg). This study validated addition of the ad hoc starter/non-starter culture for PDO Pecorino cheese production., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In recent years, the market for minimally processed fruit has increased. Fresh-cut fruits are characterized by a short shelf life due to the processing phases that accelerate the ripening courses. The aim of this work is to analyze the effect on the quality of fresh-cut Hayward kiwis of three different edible coatings based on (1) Aloe vera gel, (2) Aloe vera gel + hydroxypropyl methylcellulose and (3) Aloe vera gel + lemon essential oil. Fruit firmness, weight loss, color, soluble solids content, titratable acidity, microbial load and sensory analysis were evaluated as fresh after 2, 4, 7 and 10 days. Aloe vera gel and Aloe vera gel + lemon essential oil maintained the best values, as they acted as a barrier to gas exchange and further reduced the microbial load. These results were confirmed by sensory analysis: Aloe vera gel + hydroxypropyl methylcellulose does not alter the natural taste of kiwi slices, Aloe vera gel + lemon essential oil gives the characteristic taste of lemon essential oil and Aloe vera gel gives an herbaceous taste. The Aloe vera gel, in combination with these additives, maintains the ability to preserve the quality of fresh-cut kiwifruit.

In the present work the microbial dynamics in wheat kernels were evaluated over time. The main aim of this research was to study the resistance of lactic acid bacteria (LAB) and yeasts associated to unprocessed cereals used for bread making during long term conservation. To this purpose four Triticum durum Desf. genotypes including two modern varieties (Claudio and Simeto) and two Sicilian wheat landraces (Russello and Timilia) were analysed by a combined culture-independent and -dependent microbiological approach after one, two or three years from cultivation and threshing. DNA based MiSeq Illumina technology was applied to reveal the entire bacterial composition of all semolina samples. The samples showed a different distribution of bacterial taxa per variety and time of storage. The groups mostly represented were Stenotrophomonas, Pseudomonas, Erwinia, Delftia and Sphingomonas genera, Enterobacteriaceae and Oxalobacteriaceae families, and Actinobacteria phylum. Among LAB, only Enterococcus genus was detected barely in a single sample (Simeto stored for one year) by the next generation sequencing, indicating that LAB remained unassigned or their abundances were below 0.1% or their DNAs were rendered inaccessible. Plate counts showed consistent differences in relation to genotypes and duration of storage, with the highest levels found for total mesophilic microorganisms detected up 6.8 Log CFU/g. Colonies of presumptive sourdough LAB were detected only in a few samples. Cocci constituted the major group of LAB in almost all samples. Following the enrichment procedure, almost all samples were characterised by the presence of acidifying microorganisms. All isolates collected before and after enrichment represented 28 different strains belonging to 10 species of Enterococcus, Lactobacillus and Pediococcus genera. The most resistant species during aging were Enterococcus faecium, Enterococcus durans, Lactobacillus brevis, Lactobacillus pentosus and Lactobacillus paracasei demonstrating that lactobacilli and enterococci are able to overcome the stressing conditions represented by cereal storage better than other LAB genera commonly found associated to cereals after harvest. Yeast community included mostly species with no interest in bread making., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The main hypothesis of this work was to evaluate the presence of lactic acid bacteria (LAB) intrinsically resistant to plant essential oils in sage (Salvia officinalis L.) and laurel (Laurus nobilis), for future applications in functional cheese production by addition of aromatic herbs. The effect of the drying process on the viability of LAB was evaluated with three biomass densities (3, 4 and 5 kg/m2). The drying densities did not affect weight loss, but influenced the levels of LAB of sage and laurel. A total of 10 different strains of Enterococcus faecium, Enterococcus mundtii, Enterococcus raffinosus and Leuconostoc mesenteroides were identified from laurel, while sage did not host any LAB species. In particular, L. mesenteroides was the only species sensitive to the heat treatment. Only five strains, all enterococci, were resistant to at least one antibiotic, even though no strain showed gelatinase or haemolytic activity. The investigation on the technological traits useful in cheese making demonstrated that all LAB can be considered non starter LAB, because they were characterized by a slow acidification capacity (the pH was still above 6.00 after 3 d) and a very limited autolysis (the maximum decrease of the optical density at 599 nm was barely 0.2)., (© FEMS 2020.)

The present work was carried out to evaluate the microbiological and physicochemical composition of salamis produced with the meat of beef, horse, wild boar and pork. Salami productions occurred under controlled laboratory conditions to exclude butchery environmental contaminations, without the addition of nitrate and nitrite. All trials were monitored during the ripening (13 °C and 90% relative humidity) extended until 45 d. The evolution of physicochemical parameters showed that beef and pork salamis were characterized by a higher content of branched chain fatty acids (FA) and rumenic acid than horse and wild boar salamis, whereas the last two productions showed higher values of secondary lipid oxidation. Plate counts showed that lactic acid bacteria (LAB), yeasts and coagulase-negative staphylococci (CNS) populations dominated the microbial community of all productions with Lactobacillus and Staphylococcus as most frequently isolated bacteria. The microbial diversity evaluated by MiSeq Illumina showed the presence of members of Gammaproteobacteria phylum, Moraxellaceae family, Acinetobacter, Pseudomonas, Carnobacterium and Enterococcus in all salamis. This study showed the natural evolution of indigenous fermented meat starter cultures and confirmed a higher suitability of horse and beef meat for nitrate/nitrite free salami production due to their hygienic quality at 30 d., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Pistachio powder was added to flour or semolina to evaluate its contribution to increase the amount of lysine in bread. Bread production was carried out by sourdough technology using a selected 3-species (Lactobacillus sanfranciscensis/Leuconostoc citreum/Weissella cibaria) lactic acid bacterial (LAB) starter culture. All sourdoughs were subjected to a long-time fermentation (21 h) and showed levels of LAB around 10 9  CFU/g, indicating the suitability of pistachio powder for lactic fermentation. Yeasts were also detected, in particular in semolina trials. MiSeq Illumina technology was applied to investigate the bacterial composition of sourdoughs evidencing a different distribution of LAB species among the trials with Lactobacillus as major LAB group in almost all sourdoughs. Physicochemical parameters were comparable among the trials. After baking, pistachio powder was found not to influence the height of the breads, but pistachio breads were more firm than control breads. Color of the breads, void fraction and cell density, were influenced by pistachio powder. The amount of lysine increased consistently thanks to pistachio supplementation which also determined a higher presence of o-xylene, p-cymene and limonene and the appearance of α-pinene and 1-octen-3-ol in breads. Sensory tests showed the best appreciation scores for the breads produced with flour and pistachio powder., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The present work was developed to select lactic acid bacteria (LAB) to be used as starter cultures in functional cheese production. The indigenous milk LAB populations were isolated from fermented raw ewes' milks (four bulks) added with 0.5 mg/mL of nine polyphenols commonly found in winery by-products. After 48 h of fermentation, all milks were characterized by an increase of LAB levels of about 3-4 Log cycles. All different colonies were purified and characterized for the main physiological and biochemical traits and then differentiated genetically at strain level and identified. Ten species belonging to the LAB genera Lactobacillus, Streptococcus, Enterococcus, Leuconostoc and Lactococcus were identified. Only Lactococcus lactis and Leuconostoc mesenteroides strains were evaluated for the technological traits including acidification and autolytic kinetics, diacetyl formation, exopolysaccharide production and generation of antimicrobial compounds. A total of four strains (Mise36, Mise94 Mise169 and Mise190) belonging to Lc. lactis displayed potential for production of cheeses containing grape polyphenols., (© FEMS 2020.)

A novel dairy product, namely "chocolate cheese", was produced with two typical Sicilian food products: Pecorino cheese, processed from ewe's milk, and Modica chocolate. The cheese, manufactured with 0%, 5%, 10%, and 15% (w/w) solid or molten chocolate, was evaluated after 0, 2, 4, and 6 weeks of vacuum storage for its nutritional and health properties. The addition of chocolate reduced the pH, protein, fat, and ash; the addition of 5% or 10% molten chocolate reduced hardness (N/mm 2 ). The addition of either solid or molten chocolate resulted in a slight increase (P < 0.1038) in the total polyphenol content, a higher oleic acid content, and less oxidative stability. The microbiological profile showed that the total mesophilic count and the number of mesophilic coccus lactic acid bacteria (LAB) were approximately equal (about 10 8 CFU/g) in all cheese. The survival of the microorganisms was affected by both the chocolate added and the storage time. Chocolate cheese stored for 6 weeks had less Enterobacteriaceae than control cheese, whereas yeasts were detected at higher cell densities in the former cheese. Filamentous fungi were undetectable in some cheese. Differences were also observed in the number of mesophilic rod LAB, which increased progressively over time in all cheese, and in Enterobacteriaceae, yeasts, and filamentous fungi, which decreased during storage. Descriptive and hedonic sensory tests and principal component analysis showed that fresh cheese and cheese stored for 2 weeks, including 5% molten chocolate, were the most preferred by evaluators. Based on these results, chocolate cheese has the potential to be appreciated in the market for its nutritional, health, and sensory properties. PRACTICAL APPLICATION: Chocolate cheese, made by combining two typical Sicilian foods, Pecorino cheese and Modica chocolate, is proposed as a novel dairy product. The highest sensory acceptance was obtained with the addition of 5% molten chocolate and storage for 2 weeks. Given its improved antioxidant properties, healthier fat, and sensory properties, chocolate cheese has the potential to be appreciated in the market, especially by young consumers., (© 2020 Institute of Food Technologists®.)

The addition of active compounds to enhance the functional properties of foods is a quite common practice. Recently, bread became one of the target foods to incorporate functional ingredients such as those deriving from Opuntia spp. So far, only Opuntia ficus-indica cladodes in powder has been tested. The addition of fresh O. ficus-indica mucilage (in substitution to water) did not influence the biological leavening of the doughs. The resulting breads showed a biological role of the cactus mucilage, because their antioxidant activity was higher than that of control wheat bread. The sensory analysis indicated a general appreciation of the breads enriched with O. ficus-indica mucilage by the judges. The inclusion of fresh cactus mucilage in bread production might increase the dietary antioxidant intake due to its daily worldwide consumption., (Copyright © 2019 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

This work was carried out to pursue a double objective: to improve the hygienic safety of cheeses produced from raw ewes' milk; and to produce a new typology of raw ewes' milk through the application of "Grana" technology for which the name "Gran Ovino" was chosen. With this in mind, raw milk from an individual farm was transformed under controlled conditions at a dairy pilot plant. The production technology included the partial skimming of the evening and morning milk mixture by cream surfacing and the addition of a natural whey starter cultures (NWSC) prepared with four selected Streptococcus thermophilus strains (PON6, PON244, PON261 e PON413). Ten microbial groups were investigated by plate counts from raw milk until ripened cheeses. Lactic acid bacteria (LAB) were in the range 10 4 -10 5  CFU/ml before NWSC addition. After curdling, this group increased by 3 log cycles and was counted at 10 6  CFU/g after curd cooking. A rapid pH drop (to 6.05) was registered after almost 3 h from NWSC addition. The levels of members of the Enterobacteriaceae family were at about 10 3  CFU/ml in raw milk and decreased after curd cooking to 1 log cycle. A similar behavior was shown by the other undesired microbial groups and a complete disappearance of staphylococci was registered. The microbiological counts of 9-month ripened cheeses showed the dominance of LAB and undetectable levels of the undesired bacteria. MiSeq Illumina was applied to better investigate the bacterial composition of ripened cheeses and this technique evidenced that the majority of OTUs belonged to Lactobacillus and Streptococcus genera. The final cheeses were characterized by 67.65% dry matter of which 41.85% of fats and 47.02% of proteins. The main cheese fatty acids were palmitic, oleic and myristic acids and the saturated fatty acids/unsaturated fatty acids ratio was 2.17. Forty-one volatile compounds, including acids, esters, ketones, alcohols, aldehydes, phenols and one terpene were emitted from the cheese. Sensory evaluation showed a general appreciation for the new cheese product by judges., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Durum wheat bran (DWB) is a by-product mostly used in feeding ruminants, contributing to decrease in the utilization of feeds suitable as foods for human consumption, thus improving the sustainability of livestock production. However, the potential benefits of DWB, due to its content in phenolic acids, mainly consisting of ferulic acid with antioxidant properties, have not been well clarified yet. Accordingly, in this experiment, 36 lactating cows divided into three groups received, over a period of 100 days, one of three concentrates including DWB at 0% (DWB0), 10% (DWB10), or 20% (DWB20). The concentrates were formulated to be isoproteic and isoenergetic and, to balance the higher fiber content of the concentrates with DWB, the hay in the diets was slightly reduced. During the trial, the group feed intake and the individual milk production were monitored, and cheese was made with bulk milk from each group. Milk yield and microbiological characteristics of milk and cheese were similar among groups, indicating no DWB effect on cows performance and fermentation process. Milk from DWB20 group resulted slightly higher in casein and curd firmness (a 2r ). In cows fed DWB, the higher polyphenol intake was responsible for higher blood contents of these bioactive compounds, that seemed to have contributed in reducing the level of reactive oxygen metabolites (ROMs), which were higher in DWB0 cows. DWB20 cheeses showed a higher polyphenol content, lower number of peroxides, and higher antioxidant capacity than DWB0 cheeses. DWB20 and DWB10 diets resulted less expensive. In addition, the DWB20 group showed the best indexes heFCE (human edible feed conversion efficiency = milk/human edible feed) and NFP (net food production = milk - human edible food), expressed as crude protein or gross energy. In conclusion, the DWB fed to dairy cows at 12% of diet dry matter (DM) can lead to benefits, such as the improvement of oxidative status of cows, milk quality, shelf-life, and functional properties of cheese, and might contribute to reduce the feeding cost and limit the human-animal competition for feeding sources.