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Strawberry fruit is a nonclimacteric fruit and is one of the most consumed berries in the world. It is characterized by high levels of vitamin C, folate, vitamin E, β-carotene, and phenolic constituents as well asanthocyanins that are strictly related to health benefits. Strawberries are highly perishable fruit with a very short postharvest life due to their susceptibility to mechanical injury, rapid texture softening, physiological disorders, and infection caused by several pathogens (yeast and mold) that can rapidly reduce fruit quality. The aim of the present study was to evaluate the effect of the application of Opuntia ficus-indica mucilage in combination with ascorbic acid, as edible coating, on quality, sensorial parameters, and microbiological characteristics of strawberry fruit during cold storage at 4 ± 0.5°C and 85% RH. Strawberries were characterized by a linear increase of weight loss during the storage at 4°C that was significantly higher (+11.3% on average) in the uncoated strawberries. The coating affected the ascorbic acid content of the strawberries that increased by 36.0% in coated strawberries; total soluble solid content and color of the strawberries were only affected by storage. Visual quality and sensorial analysis recorded higher scores in the coated samples at the end of the cold storage period. Furthermore, the mucilage coating did not negatively affect the natural taste of strawberries. The application of O. ficus-indica gel-based edible coating in combination with ascorbic acid, although not able to inhibit the microbial growth, limited significantly their development in coated strawberry fruits. Our results suggest that Opuntia mucilage plus 5% ascorbic acid could be a useful biochemical way of maintaining strawberry fruit quality and extending their postharvest life.

The agricultural practices applied in pre-harvest greatly influence the presence and the levels of microorganisms in fresh produce. Among these, Listeria monocytogenes represents one of the most lethal foodborne pathogens associated with vegetables. The main hypothesis of this work is that bacteriocin producer Enterococcus mundtii strains can be effective against L. monocytogenes in soil. To this purpose, bacteriocin production by E. mundtii WFE3, WFE20 and WFE31, three strains showing a strong bacteriocin activity in terms of inhibitory power and inhibition spectra, was evaluated in sterile extracts from agricultural soil and peat moss, in organic nutrient solution (ONS) and mineral nutrient solution (MNS). ONS supernatants from E. mundtii WFE3 showed the highest inhibition of the strain L. monocytogenes ATCC 19114. Thus, this strain [104 colony forming units (CFU) g−1 dw] was co-inoculated with E. mundtii WFE3 (106 CFU g−1 dw) in the sterile extracts and solutions. A general increase of E. mundtii cell densities and the contemporary decrease of the levels of L. monocytogenes were observed, particularly in ONS. This solution was used to amend autoclaved soil to test in vivo the competition among bacteriocin producer and sensitive strain. L. monocytogenes decreased of almost 1.5 log CFU when E. mundtii was added to soil. The test was also carried out with basil plants showing that the anti-Listeria effect of E. mundtii is exerted during the very first days from inoculation. After 2 days, the levels of NO3−-N in soil increased for all trials, while the concentrations of ammonium and α-amino N decreased. The lowest concentrations of NH4+-N were found in presence of L. monocytogenes. At harvest, the plants were analysed for the presence of E. mundtii and L. monocytogenes, but none of the two bacterial species deliberately added to soil was transferred to plants. Although a biocontrol based on bacteriocin application in soil has to be properly set, this study provides useful insight for the future development of chemical-free agricultural strategies.

Cactus pear (Opuntia ficus-indica (L.) Mill.) is a non-climacteric fruit with a relatively short postharvest life span, being very sensitive to water loss, darkening and decay. Cactus pear is a spiny fruit, and the presence of glochids limits fruit consumption and diffusion, therefore, minimally processing, as well as peel removing, could be an opportunity to improve its availability, consumption, and diffusion in national and international markets. In this study, cactus pear minimally processed fruits were treated with a mucilage-based coating extracted from Opuntia ficus-indica cladodes and stored at 5 °C for 9 days. The effect of mucilage edible coating on the postharvest life, qualitative attributes, and nutraceutical value of fruit were evaluated by colors, firmness, total soluble solids content, titratable acidity, ascorbic acid, betalains and DPPH (2,2-diphenyl-1-picrylhydrazyl). Results showed that mucilage-based coating improved the quality and preserves the nutraceutical value of minimally processed cactus pear fruits during storage. The edible coating was effective in maintaining fruit fresh weight, total soluble solids content, fruit firmness, ascorbic acid and betalain content, sensorial traits, and visual score. Coated fruits showed a significantly lower microbiological growth than uncoated control fruits during the entire cold storage period.

Previous investigations on pizza dough lactic acid bacteria (LAB) revealed that facultative heterofermentative species (FHS) were more represented than obligate heterofermentative species (OHS) within the Lactobacillus genus. Thus, the main hypothesis of this work was that facultative and obligate heterofermentative Lactobacillus species can impact differently the appreciation of baked pizza. The performances of different Lactobacillus, including L. sanfranciscensis, L. brevis and L. rossiae among OHS and L. plantarum, L. graminis and L. curvatus among FHS were tested in single or multiple combinations during pizza production. The values of pH, total titratable acidity and LAB levels indicated that the acidification process was almost comparable among trials. The fermentation quotient of FHS trials was above 4.0. All trials were dominated by the added LAB and for the trials with the multiple strain starter inoculums, the species found at the highest cell densities were L. sanfranciscensis, L. brevis and L. plantarum. Significant differences among pizzas were found for weight loss, colour, morphology and volatile organic compounds (VOCs). The last analysis revealed the presence of eight chemical classes with aldehydes, esters, alcohols and acids as major compounds and allowed the separation of the trials FHS and OHS. Sensory attributes were significantly different for judges and pizzas and the most relevant differences were found for crust colour, presence of bubbles, resistance to tearing, crispness and chewiness. The overall assessment reached the highest scores for the mixed culture of OHS and FHS together.

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 (Na2SeO3) 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.

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 109 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.

Adequate fruit consumption helps to prevent several chronic age-related diseases. Selenium (Se) is an elemental micronutrient with antioxidant capacity. In general, fruits and Se ingest by humans are below the recommended daily intake value. Fresh fruits are highly susceptible to deterioration during storage. Fermentation can improve the storage period, sensory profile, and bioactive compound content of foods; moreover, some lactic acid bacteria can accumulate organic Se intracellularly. In this work, microbial growth and Se accumulation by Levilactobacillus brevis CRL2051 and Fructobacillus tropaeoli CRL2034 in tropical fruit juices were evaluated. The strains could grow 1–2 log cfu/mL, alone or combined, in mango, passion fruit, and mango/passion fruit juices, although they could not completely eliminate native microorganisms in unpasteurised juices. In pasteurised trials, both strains consumed fruit carbohydrates producing lactic acid, in addition to acetic acid, and mannitol by F. tropaeoli. Both strains accumulated Se intracellularly in the fruit juices, especially (123.0 μg/L) in the passion fruit juice fermented by the mixed culture. Finally, the Fructobacillus strain increased 3.42 times the phenolic compound concentration in the mango/passion fruit juice with added Se after 24 h. The assayed LAB strains could be used for preparing functional fermented fruit beverages bio-enriched in Se.

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.

Red grape Nero d’Avola cultivar grape pomace powder (GPP) was applied during fresh ovine cheese production in order to increase polyphenol content. Before cheeses were produced, the bacteria of a freeze-dried commercial starter culture were isolated and tested in vitro against GPP. Two dominant strains, both resistant to GPP, were identified. Thestarter culture was inoculated in pasteurized ewe’s milk and the curd was divided into two bulks, one added with 1% (w/w) GPP and another one GPP-free. GPP did not influence the starter culture development, since lactic acid bacteria (LAB) counts were 109 CFU/g in both cheeses at 30 d. To exclude the interference of indigenous LAB, the pasteurized milk was analyzed, and several colonies of presumptive LAB were isolated, purified and typed. Four strains were allotted into Enterococcus and Lacticaseibacillus genera. The direct comparison of the polymorphic profiles of cheese bacteria evidenced the dominance of the starter culture over milk LAB. The addition of GPP increased cheese total phenolic compounds by 0.42 g GAE/kg. Sensory evaluation indicated that GPP-enriched cheese was well appreciated by the judges, providing evidence that GPP is a suitable substrate to increase the availability of total phenolic content in fresh ovine cheese.

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).

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.

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.

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.

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 109 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.

Pleurotus eryngii (DC.) Quél. powder was used in bread production. Three dough trials (0, 5 and 10% of mushroom) were obtained with commercial baker's yeast. P.eryngii powder was first tested against several yeast species; 10% P.eryngii trial was characterised by the highest pH and total titratable acidity. P.eryngii did not influence negatively the fermentation process, since all trials reached yeast levels of 10 8 CFUg −1 . Mushroom powder decreased bread height and softness, increased crust redness and crumb void fraction and cell density and, although the breads were scored diverse, the overall assessment was comparable. The final breads provided higher concentrations of thiamin, riboflavin and pantothenic acid than control breads and, mostly importantly, supplied biotin, cobalamin and cholecalciferol generally absent in wheat bread. P.eryngii can be cultivated on food residues. Thus, its inclusion in functional bread production represents an optimal strategy for the valorisation of food processing by-products.

This work was aimed to produce an “active” food ice to preserve its microbiological safety over time. With this in mind, ice cubes were processed with the addition of H2O2 to water before freezing. Four food ice productions were performed at the industrial level: one control trial without the addition of H2O2 (0OX) and three experimental trials obtained by adding 4, 8, and 12 mg/L of H2O2 (4OX, 8OX, and 12OX), respectively. After production, all food ice trials were artificially contaminated with 102 CFU/100 mL of water-borne pathogenic bacteria (Escherichia coli ATCC 25922, Enteroccus faecalis ATCC 29212, and Pseudomonas aeruginosa ATCC 27853) inoculated individually. Thawed ice samples were then subjected to microbiological analyses performed by the membrane filtration method and the results indicated that only trial 12OX was able to inactivate all bacteria strains. In conclusion, the addition of 12 mg/L H2O2 represents an optimal cost-effective strategy to preserve the microbiological stability of food ice even when it is improperly handled after production.

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.

Powdered mealworm (MW) and buffalo worm (BW) larvae were used to functionalize sourdough Italian-style breads. Sourdough inoculum was started with Levilactobacillus brevis, Weissella cibaria and Leuconostoc citreum. The doughs were SBS (semolina plus powdered BW larvae and sourdough) and SMS (semolina plus powdered MW larvae and sourdough) whose pHs (4.32 and 4.21, respectively) were higher than that of control (3.81). The highest fermentation quotient (lactate/acetate molar ratio) was recorded in SMS (4.46). LAB reached viable counts of about 109 CFU g−1 in almost all doughs. Insects impacted bread VOCs with dodecanal, 2.4-dodecadienal and 2-octenal-2-butyl. SBS and SMS increased the antioxidant capacity of breads by 42 and 69%, respectively. SMS decreased the glucose release in the bio-accessible fraction by 70% reducing the glycemic index of bread. Control breads were more appreciated by the sensory panel than insect-containing breads, though SMS breads were characterized by a slightly lower overall assessment.

A commercial biodegradable starch-based polymer (Mater-Bi) was activated with carvacrol to develop a biodegradable and compostable polymer to be used in food packaging. Based on previous tests, carvacrol was added at 20 % weight of foam. MB foams, with and without carvacrol, were tested for their morphological characteristics, mechanical tests and kinetics of carvacrol release under refrigerated storage conditions. Carvacrol slightly increased the porosity of the foams, induced a reduction of the compressive elastic modulus (Ecom) of foamed MB from 6 to ∼ 3.4 MPa and a decrease of the tensile elastic modulus from ∼70 MPa to ∼16.5 MPa. Carvacrol release from the foam at 4 °C was almost 7% of the initially loaded amount at the fifth hour, while 35.4 % at the end of the test (900 h). MB containing carvacrol was active against several pathogenic and spoilage bacteria in vitro. Trays made of MB containing carvacrol were put in contact with meat, fish and vegetable food systems, artificially contaminated with Pseudomonas poae 4G558 and Listeria monocytogenes 13BO. No antimicrobial effect of MB foams was registered for ham and salmon trials, while a clear inhibitory effect of carvacrol activated MB foams was observed for contaminated melon and pumpkin in which both bacteria decreased of about 1 log cycle after 3 d and completely disappeared from the 7th d of refrigerated storage. This study provided evidences on the suitability of MB foams containing 20 % carvacrol as active packaging systems for vegetables even though it negatively affected their physicochemical parameters and overall quality.

The “Spiritu re fascitrari” (SRF) is a typical Sicilian distillate obtained from the by-products of traditional process of honey production. Although some alcoholic fermentation of honey based products have been described, the present research represents the first investigation on the yeast ecology and the physico-chemical characteristics of honey by-products subjected to an alcoholic fermentation followed by distillation. All samples collected during manufacturing process were analysed for the count of total, osmophilic and osmotolerant yeasts. The honeycombs and equipment surfaces showed the presence of yeasts that was 1.7 and 1.1 Log (CFU/mL), respectively. After enrichment, yeast populations increased and a significant increase of yeasts was registered during the alcoholic fermentation (AF), reaching loads higher than 7 Log CFU/mL after day 6. A total of 2816 colonies of yeasts were isolated from the count plates and the following species were genetically identified: Lachancea fermentati, Pichia anomala, Pichia kudriavzevii, Saccharomyces cerevisiae, Wickerhamomyces anomalus, Zygosaccharomyces bailii and Zygosaccharomyces rouxii. During the spontaneous AF process, the species S. cerevisiae, Z. bailii and Z. rouxii were mainly isolated and the feed conversion ratio of sugars into ethanol was about 53%; high contents of acetic acid and glycerol were also found. The highest concentrations of volatile organic compounds (VOCs) were registered for esters, alcohols and aldehydes (346.55, 331.041 and 13.65 μg/L, respectively). Many VOCs identified as “specific floral markers” such as nonanal and 1-hexanol, 1-octanal and linalool oxide were found. Although more studies are needed, our results suggested that the S. cerevisiae strains isolated in this study must be evaluated in situ for their potential to act as starters for the continuous production of SRF. This because these strains are expected to drive the fermentation process reducing the risk of off-odour and off-flavour formation.

Microbiological, nutritional and bioactive properties of edible powders obtained from Acheta domesticus (house cricket) and Tenebrio molitor (mealworm) were investigated. Except for the enterobacteria, viable bacteria were at a higher concentration in mealworm flour. The diversity evaluation carried out using MiSeq Illumina that mainly identified Citrobacter and Enterobacteriaceae in mealworm powder and members of the Porphyromonadaceae family in house cricket powder. Enterococci were identified and characterized for their safety characteristics in terms of the absence of antibiotic resistance and virulence. Both powders represent a good source of proteins and lipids. The fatty acid profile of mealworm powder was characterized by the predominance of the monounsaturated fatty acids and house cricket powder by saturated fatty acids. The enzymatic hydrolysis produced the best results in terms of percentage of degree of hydrolysis with the enzyme Alcalase, and these data were confirmed by SDS-PAGE electrophoresis. Furthermore, the results showed that the protein hydrolysate of these powders produces a significant antioxidant power.

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 (a2r). 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 &minus, 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.

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/mm2). The addition of either solid or molten chocolate resulted in a slight increase (P

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 104–105 CFU/ml before NWSC addition. After curdling, this group increased by 3 log cycles and was counted at 106 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 103 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.

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 evaluate the microbiological, physicochemical, and sensory characteristics of fortified pistachio breads. Pistachio powder (5% w/w) was added to flour or semolina and fermented by a commercial baker’s yeast (Saccharomyces cerevisiae). Pistachio powder did not influence the biological leavening of the doughs. The kinetics of pH and total titratable acidity (TTA) during dough fermentation showed that the leavening process occurred similarly for all trials. The concentration of yeasts increased during fermentation and reached levels of 108 CFU/g after 2 h. Pistachio powder decreased the height and softness of the final breads and increased cell density of the central slices. The amount of lysine after baking increased in pistachio breads and this effect was stronger for semolina rather than flour trials. Sensory evaluation indicated that fortified breads processed from semolina were those more appreciated by the judges. This work clearly indicated that the addition of pistachio powder in bread production represents a promising strategy to increase the availability of lysine in cereal-based fermented products.

In this work, the feasibility of antibacterial biopolymeric foams containing carvacrol (CRV) for potential food packaging applications was investigated. Sodium bicarbonate (SB) was chosen as foaming agent and a commercial biodegradable polymer, Mater-Bi® (MB), as a matrix. MB/SB and MB/SB/CRV systems were prepared by melt mixing and the foaming process was conducted in a laboratory press. The influence of foaming agent and antibacterial additive content was investigated. The foamed samples were characterized through morphological analysis, mechanical tests and measurements of CRV release kinetics. Moreover, a mathematical model, i.e. power law model, was used to fit the release data in order to investigate the release mechanism. The antibacterial activity of foams containing CRV was tested in vitro against pro-technological, spoilage and pathogenic bacteria applying the disc diffusion method. The results showed that by using 3% of SB it is possible obtain foams with a porosity of about 40%. The presence of carvacrol, strongly influenced the properties of the biopolymeric foams but at the higher content ensured antibacterial activity of the samples.

Listeria monocytogenes is a pathogen frequently found in dairy products, and its growth is difficult to control. Bacteriocinlike inhibitory substances (BLIS), produced by lactic acid bacteria (LAB), having proven in vitro anti-Listeria activity, could provide an innovative approach to control L. monocytogenes; however, this application needs to be evaluated in vivo. In this study, twenty LAB strains isolated from different Sicilian dairy environments were tested for control of growth of L. monocytogenes in three different experimental trials. First, raw and UHT milk were inoculated with LAB strains alone, and LAB strains mixed with L. monocytogenes. Second, mini-cheeses containing LAB and/or L. monocytogenes were produced. Third, two traditional Sicilian cheeses inoculated with a multi-strain LAB mixture combined with L. monocytogenes were produced. The addition of BLIS produced by LAB to milk and in mini-cheese production was unable to inhibit the growth of L. monocytogenes. However, an anti-Listeria effect was observed in the Pecorino Siciliano cheeses, where, after 15 days of ripening, the cheeses with added LAB had fewer L. monocytogenes compared to the control cheeses with no added LAB, while in the Vastedda della valle del Belìce cheeses, the multi-strain LAB mixture completely prevented the growth of L. monocytogenes.

The main aim of this research was to investigate the biopreservative effects of lemon essential oil (EO) micro-emulsions on salted sardines. The experimental design included two experimental trials, SR1 and SR2 carried out with 25 ml of lemon EO micro-emulsion at 0.3 and 1.0% (v/v), respectively, and a control trial performed without EO addition. Chemical analyses on salted sardines inoculated with the EOs clearly showed a substantial persistence of several volatile organic compounds (VOCs) belonging to groups of monoterpene hydrocarbons, oxygenated monoterpenes and sesquiterpene hydrocarbons derived from EOs during the entire period of ripening. In particular, the molecules mostly represented were limonene, p-cymene and (beta-pinene. Immediately after the addition of EOs, the concentrations of all microbial groups decreased. The presence of Enterobacteriaceae, staphylococci and rod lactic acid bacteria (LAB) observed in the trials SR1 and SR2 was significantly lower than that registered for the control. trial during the entire period of monitoring. Furthermore, the addition of EOs determined a lower accumulation of histamine in sardines compared to those of the control trial. The highest scores of sensory evaluation were registered for flavour and overall acceptability of the experimental trials in presence of EOs. On the basis of the increasing interest toward novel food preservatives, we conclude that the use of EOs to produce salted fishes represents a valid strategy to improve safety and sensory characteristics of salted sardines. This work has also economic implications, since the flavour improvement due to the addition of lemon EOs might increase the consumption of sardines by regular and new consumers. (C) 2016 Elsevier Ltd. All rights reserved.

The main objective was to set up a methodology to improve the high volume production of green table olives, cv. Nocellara del Belice. Lactobaccillus pentosus OM13 was applied during three different industrial processes of table olives as follows: trial one (IOP1) was subjected to an addition of lactic acid until a brine level of pH 7.0 was reached; trial two (IOP2) subjected to same addition of lactic acid as in trial one plus nutrient adjuvant; and trial three (IOP3) subjected to same addition of lactic acid as in trial one, but with the strain L. pentosus OM13 acclimatized in brine for 12 h before inoculation. These trials were compared against two untreated controls (spontaneously fermented and addition of L. pentosus OM13 only). Within the third day of fermentation, the pH of the brines decreased significantly, reaching pH 4.85 for trial three, pH 5.15 for trial two, and pH 5.92 for trial one. The pH of both controls decreased more slowly, and had values below pH 5.0 only after the fifteenth day of fermentation (control one) and the sixty-fifth day of fermtation (control two). Trial three reached the highest lactic acid bacteria (LAB) concentration on the third day of fermentation. After six days of fermentation, all trials showed similar values of LAB counts that were significantly higher compared to control number one. The result from genotypic identification showed that L. pentosus OM13 was the most frequently isolated in the inoculated trials. Lactobacillus plantarum, Lactobacillus coryniformis and Pediococcus pentosaceous were also detected at very low concentrations. Homoguaiacol, 2-butanol, 4-ethylphenol, phenylethyl alcohol and 4-ethylphenol were the volatile organic compounds detected at the highest levels in all experimental trials. Trial three showed a higher concentration of squalene that was not detected in other trials. The highest sensory scores of green olive aroma and overall satisfaction were found for all experimental olives, especially for those of trial one and trial two, that differed significantly from the untreated controls. This study provides evidence that the addition of lactic acid, nutrient adjuvants and, most importantly, the acclimatization of LAB cells significantly shortens the acidification process of olive brine, and improves safety and sensory quality. Shorter acidification processes result in a more rapid transformation of table olives, with reduced commodity loss and lower costs of production compared to conventional manufacturing protocols.

Accumulation of biogenic amines (BAs) in cheese and other foods is a matter of public health concern. The aim of this study was to identify the enzyme activities responsible for BA degradation in lactic acid bacteria which were previously isolated from traditional Sicilian and Apulian cheeses. The selected strains would control the concentration of BAs during cheese manufacture. First, 431 isolates not showing genes encoding the decarboxylases responsible for BA formation were selected using PCR-based methods. Ninety-four out of the 431 isolates degraded BAs (2-phenylethylamine, cadaverine, histamine, putrescine, spermine, spermidine, tyramine, or tryptamine) during cultivation on chemically defined medium. As shown by random amplification of polymorphic DNA-PCR and partial sequencing of the 16S rRNA gene, 78 of the 94 strains were Lactobacillus species ( Lactobacillus casei , Lb. fermentum , Lb. parabuchneri , Lb. paracasei , Lb. paraplantarum , and Lb. rhamnosus ), Leuconostoc species ( Leuconostoc lactis and Ln. mesenteroides ), Pediococcus pentosaceus , Lactococcus lactis , Streptococcus species ( Streptococcus gallolyticus and S. thermophilus ), Enterococcus lactis , and Weissella paramesenteroides . A multicopper oxidase-hydrolyzing BA was purified from the most active strain, Lb. paracasei subsp. paracasei CB9CT. The gene encoding the multicopper oxidase was sequenced and was also detected in other amine-degrading strains of Lb. fermentum , Lb. paraplantarum , and P. pentosaceus . Lb. paracasei subsp. paracasei CB9CT and another strain (CACIO6CT) of the same species that was able to degrade all the BAs were singly used as adjunct starters for decreasing the concentration of histamine and tyramine in industrial Caciocavallo cheese. The results of this study disclose a feasible strategy for increasing the safety of traditional cheeses while maintaining their typical sensorial traits. IMPORTANCE Because high concentrations of the potentially toxic biogenic amines may be found in traditional/typical cheeses, the safety of these food items should be improved. Lactic acid bacteria selected for the ability to degrade biogenic amines may be used during cheese making to control the concentrations of biogenic amines.

The main hypothesis of this study was that the autochthonous lactic acid bacteria (LAB) selected for their dairy traits are able to stabilize the production of PDO (Protected Denomination of Origin) Pecorino Siciliano cheese, preserving its typicality. The experimental plan included the application of a multi-strain lactic acid bacteria (LAB) culture, composed of starter (Lactococcus lactis subsp. lactis CAG4 and CAG37) and non starter (Enterococcus faecalis PSL71, Lactococcus garviae PSL67 and Streptococcus macedonicus PSL72) strains, during the traditional production of cheese at large scale level in six factories located in different areas of Sicily. The cheese making processes were followed from milk to ripened cheeses and the effects of the added LAB were evaluated on the microbiological, chemico-physical and sensorial characteristics of the final products. Results highlighted a high variability for all investigated parameters and the dominance of LAB cocci in bulk milk samples. The experimental curds showed a faster pH drop than control curds and the levels of LAB estimated in 5-month ripened experimental cheeses (7.59 and 7.27 Log CFU/g for rods and cocci, respectively) were higher than those of control cheeses (7.02 and 6.61 Log CFU/g for rods and cocci, respectively). The comparison of the bacterial isolates by randomly amplified polymorphic DNA (RAPD)-PCR evidenced the dominance of the added starter lactococci over native milk and vat LAB, while the added non starter LAB were found at almost the same levels of the indigenous strains. The sensory evaluation showed that the mixed LAB culture did not influence the majority of the sensory attributes of the cheeses and that each factory produced cheeses with unique characteristics. Finally, the multivariate statistical analysis based on all parameters evaluated on the ripened cheeses showed the dissimilarities and the relationships among cheeses. Thus, the main hypothesis of the work was accepted since the quality parameters of the final cheeses were stabilized, but all cheeses maintained their local typicality.

In this work, an ad hoc method to identify and quantify polyphenols from caper berries was developed on high-performance liquid chromatography/electrospray ionisation source/mass spectrometry (HPLC-ESI-MS). The method was applied during fermentation carried out with Lactobacillus pentosus OM13 (Trial S) and without starter (Trial C). A total of five polyphenols were identified. All samples contained high concentrations of rutin. Epicatechin was found in untreated fruits, on the contrary quercetin was detected during fermentation. Trial S was characterised by a more rapid acidification and lower levels of spoilage microorganisms than Trial C. L. pentosus dominated among the microbial community of both trials and the highest biodiversity, in terms of strains, was displayed by Trial C. Aureobasidium pullulans was the only yeast species found. The analytical method proposed allowed a high polyphenolic compound recovery from untreated and processed caper berries in short time. The starter culture reduced the bitter taste of the final product.

The essential oils (EOs) of green seeds from Daucus carota subsp. maximus growing wild in Pantelleria Island (Sicily, Italy) were characterized. EOs were extracted by steam distillation, examined for their inhibitory properties against food-borne Gram-positive and Gram-negative bacteria and analyzed for the chemical composition by gas chromatography (GC) and mass spectrometry (MS). Undiluted EOs showed a large inhibition spectrum against Gram-positive strains and also vs. Acinetobacter spp. and Stenotrophomonas maltophilia. The minimum inhibition concentration (MIC) was in the range 1.25 – 2.50 μl/ml for the most sensitive strains. The chemical analysis indicated that D. carota subsp. maximus EOs included 34 compounds (five monoterpene hydrocarbons, six oxygenated monoterpenes, 14 sesquiterpene hydrocarbons, four oxygenated sesquiterpenes, camphorene and four other compounds), accounting for 95.48% of the total oil, and that the major chemicals were carotol, β-bisabolene, and isoelemicin.

To evaluate the efficacy of lactic acid bacteria (LAB) to improve the hygienic safety of a traditional raw milk cheese, the raw ewes’ milk protected denomination of origin (PDO) Pecorino Siciliano cheese was used as a model system. Different Pecorino Siciliano curds and cheeses were used as sources of autochthonous LAB subsequently used as starter and non-starter LAB. These were screened for their acidification capacity and autolysis. Starter LAB showing the best performance were genotypically differentiated and identified: two strains of Lactococcus lactis subsp. lactis were selected. From the nonstarter LAB, Enterococcus faecalis, Lactococcus garvieae and Streptococcus macedonicus strains were selected. The five cultures were used in individual or dual inocula to produce experimental cheeses in a dairy factory for which production was characterised by high numbers of undesirable bacteria. At 5- month of ripening, the experimental cheeses produced with LAB were characterised by undetectable levels of enterobacteria and pseudomonads and the typical sensory attributes.