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[Display omitted] • Fermentation as 'green technology' to generate high added-value products from by-products. • Pâté Olive Cake as a matrix potentially suitable for formulation of new functional foods. • Effect of fermentation on bioactive molecules, volatile organic and biological assays. In this study, the pâté olive cake (POC), a by-product of the olive oil industry, was subjected to fermentation in a bioreactor using three microbial strains, Lactiplantibacillus plantarum , Wickerhamomyces anomalus and Candida boidinii, previously isolated from fermented table olive brines. Chemical, microbiological and molecular analyses were carried out at the beginning and at the end of fermentation. The lowest pH value (4.09) was reached after 10 days in sample inoculated with C. boidinii. Microbiological analyses exhibited the dominance of yeasts throughout the whole process (from 5.5 to 7.80 Log 10 CFU/g), as confirmed by PCR-DGGE analysis. The microbial cultures affected both phenolic and volatile organic compound profiles. Moreover, the POC samples treated with different microbial strains were investigated for biological assays. The sample fermented with W. anomalus showed the greatest diffusion speed of transepithelial transport through Caco-2 cell, the highest inhibitory activity towards the tested cyclooxygenases and the highest antioxidant activity. [ABSTRACT FROM AUTHOR]

This work investigates the effects of different combinations of selected lactic acid bacteria strains on Lactobacillus species occurrence, on safety and on sensory traits of natural green table olives, produced at large factory scale. Olives belonging to Nocellara Etnea cv were processed in a 6% NaCl brine and inoculated with six different bacterial cultures, using selected strains belonging to Lactobacillus plantarum, Lactobacillus paracasei and Lactobacillus pentosus species. The fermentation process was strongly influenced by the added starters and the identification of lactic acid bacteria isolated throughout the process confirms that L. pentosus dominated all fermentations, followed by L. plantarum , whereas L. casei was never detected. Pathogens were never found, while histamine and tyrosine were detected in control and in two experimental samples. The samples with the lowest final pH values showed a safer profile and the most appreciated sensory traits. The present study highlights that selected starters promote prevalence of L. pentosus over the autochthonous microbiota throughout the whole process of Nocellara Etnea olives. [ABSTRACT FROM AUTHOR]

In this study, different microbial strains, as single and mixed-cultures, were used to ferment the pâté olive cake (POC), a by-product of olive oil processing. In particular, strains belonging to Candida boidinii, Wickerhamomyces anomalus and Lactiplantibacillus plantarum were used. The fermentation was carried out on diluted (3:2) POC without and with glucose (2% w/v) addition. Furthermore, phenolic compounds were monitored during fermentation in POC added with glucose differetly inoculated and the microbial community, at the end of fermentation, was evaluated by Next Generation Sequencing (NGS) techniques. Data highlighted that inoculated samples showed an hydroxytyrosol content higher than the un-inoculated controls. In particular, during fermentation the sample inoculated with C. boidinii, both in single and in mixed culture together with L. plantarum, increased the hydroxytyrosol content by 275 and 261 mg/L, respectively, after 8 days, to reach the highest content at the end of fermentation. Metagenomic analysis revealed a low abundance of 16S ribosomal RNA genes and fungal ITS in all samples at any sampling times. Furthermore, at the end of fermentation, all samples exhibited a different bacterial community with a decrease in acetic acid bacteria and an increase in Lactobacillaceae biodiversity. Finally, no effect was detected in any samples on fungal metagenomic profile, where Dipodascus geotrichum was found dominant both at initial and final fermentation. In conclusion, the present study confirmed that selected cultures can drive the fermentation and have an impact on the phenolic profile. [ABSTRACT FROM AUTHOR]

Abstract: To investigate Listeria innocua fate in fresh cut mixed leafy salads, packaged both in ordinary (OA) and modified atmosphere (MA), the type strain DSM 20649 was inoculated and its behaviour was monitored by conventional and molecular approaches, during storage at 4°C. Results indicated that: fresh cut salads packaged under MA (at initial conditions of 6% CO2 and 3% O2) supported the growth of Listeria spp.; conventional plating technique could not detect L. innocua even into inoculated samples, whereas; PCR–DGGE analysis showed that the L. innocua became one of the dominant species in samples packaged in MA, starting from the 3rd day of storage. The study confirms that modified atmosphere has to be applied together with other preservative techniques in order to assure the inhibition of pathogenic micro-organisms in fresh cut vegetables. [Copyright &y& Elsevier]

Abstract: Traditional artisanal Pecorino Siciliano (PS) cheeses, and two experimental PS cheeses were manufactured using either raw or pasteurised ewes'' milk with the addition of starter cultures. The bacterial diversity and dynamics of the different cheese types were evaluated both by culturing and characterisation of isolates, and a culture-independent approach based on the 16S ribosomal RNA (rRNA) gene. Following cultivation, artisanal and experimental cheese types showed similar microbial counts, and isolates belonging to Lactococcus lactis, Streptococcus thermophilus, Enterococcus faecalis and Leuconostoc mesenteroides were identified by phenotypic characterisation and comparison of the restriction fragment length polymorphism (RFLP) of the 16S rRNA gene to that of reference species. The culture-independent fingerprinting technique PCR and denaturing gradient gel electrophoresis (DGGE) of V6 to V8 regions of the 16S rRNA gene of samples taken during artisanal PS cheese manufacture, from raw milk to the ripened cheese, indicated relevant shifts in the microbial community structure. The dominance of Streptococcus bovis and Lactococcus lactis species in the traditional artisanal PS was revealed by 16S rRNA gene sequencing. Comparison of DGGE profiles of samples from milk to ripened cheese, derived from artisanal procedure and the two experimental PS cheeses during production showed similar trends with the presence of intense bands in common. Nevertheless, the profiles of several artisanal cheeses from different farms appeared more diverse, and these additional species are probably responsible for the generally superior flavour and aroma development of traditional PS cheese. [Copyright &y& Elsevier]

Microbiological safety of green table olives from different cultivars, prepared by both the Spanish-style and biological methods and fermented with starter cultures of lactic acid bacteria (Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus pentosus), was investigated. The fermentation process was monitored by measuring pH values, titratable acidities, and growth of lactic acid bacteria over time. During fermentation, lactic acid bacteria and yeasts were major microbial populations. Microbiological safety was evaluated by analysis for Listeria monocytogenes with the use of an enrichment method during storage (from 55 days to 18 months). Results demonstrated that L. monocytogenes can survive and grow in green table olives. L. monocytogenes was found in one of the commercial (thermally treated) samples analyzed and in all samples older than 2 months, irrespective of olive cultivar, lactic acid bacteria starter used, pH and titratable acidity of brine samples, or treatment applied. [ABSTRACT FROM AUTHOR]

This study investigated the phenotypic and genotypic characteristics of lactic acid bacteria in naturally fermented green olives, collected from different areas of Sicily. Both classical biochemical tests and PCR/Restriction Fragments Length Polymorphism (RFLP) of 16S rDNA were used to characterize the isolates. The identity of the isolates was obtained by the partial sequencing analysis of the 16S rDNA. The BioMerieux software assigned the 13 heterofermentative strains to the Lactobacillus brevis species; 24 homofermentative strains were classified as Lactobacillus casei and the remaining 11 homofermentative lactobacilli were identified as Lactobacillus plantarum. The rapid ID 32 STREP test identified coccal-shaped strains as Enterococcus faecium species. The PCR/RFLP analysis showed a remarkable bacterial heterogeneity within the isolates. The 16S rDNA partial sequencing did not confirm biochemical identification, revealing a strong dominance of isolates belonging to the L. casei species. It is noteworthy that this species has never been reported as dominant species in fermented vegetables.A combination of molecular and biochemical analysis allowed the identification of species involved in natural food fermentations. [Copyright &y& Elsevier]

Abstract: The yeast microbiota associated with naturally fermented and inoculated green table olives, differently treated in the field with non-conventional repellent and antiovipositional products in the control of Bactrocera oleae, was analysed using a combination of culture-dependent and -independent molecular fingerprinting. The routine yeast isolation gave rise to 118 strains, whose identification was performed by PCR-RFLP of the internal transcribed spacer (ITS) regions. Total DNA was extracted directly from the brine throughout fermentation by means of an experimental protocol that included the removal of Taq polymerase inhibitors. Denaturing Gradient Gel Electrophoresis (DGGE) of 26S rRNA gene PCR amplicons highlighted the yeast community. Comparison of both culture-dependent and independent methods indicated that the yeast species Saccharomyces cerevisiae, Wickerhamomyces anomalus, Candida diddensiae and Issatchenkia orientalis were dominant during fermentation despite the addition of the Lactobacillus plantarum starter used in brining. The resultant isolated species were unaffected by treatments in field, except for C. diddensiae whose growth was delayed by kaolin. [Copyright &y& Elsevier]

Graphical abstract Highlights • Constructed wetlands showed low efficiency in removing enterococci. • E. faecium was the dominant species in the UWTP system. • High prevalence of multidrug resistant enterococci was detected in all effluents. • Numerous unique pulso-types were detected. • Recurrent pulso-types showed different antibiotic resistance patterns. Abstract Enterococci isolated from different sites of an urban wastewater treatment plant (consisting of three horizontal subsurface flow constructed wetlands) were investigated. One-hundred-thirty isolates were identified at species level and tested for resistance to eleven antibiotics, by microdilution method, and their clonal relatedness was established by Sma I-PFGE analysis. Results highlighted the persistence of enterococcal population in all effluents and the dominance of E. faecium species. A high incidence of antibiotic resistance against erythromycin, chloramphenicol, rifampicin and ampicillin was observed, with 120 strains (93%) showing a multi-drug-resistance. Numerous pulso-types with a unique pattern were detected indicating a high diversity within enterococcal population. The recurrence of some pulso-types in different effluents was disclosed and, within the same pulso-types, different resistance patterns were observed. Comparing the MIC values of strains from inlet and outlet, different trends were observed, highlighting a certain variability among constructed wetlands in affecting the antibiotic resistance among enterococcal population. [ABSTRACT FROM AUTHOR]

Abstract In the present study, 110 enterococci were isolated from two Sicilian cheese types, Ragusano PDO and Pecorino Siciliano. Isolates, firstly identified by MALDI-TOF/MS and a multiplex PCR assay, were tested for susceptibility to the most relevant clinical antibiotics. Clonal relationships among isolates were evaluated by pulsed-field-gel electrophoresis (PFGE) analysis and the presence of vanA and vanB genes, in vancomycin resistant enterococci (VRE), was investigated. Overall, E. faecalis, E. durans (35% for each species) and E. faecium (28%) were the major identified species. Different occurrence between cheese types was revealed. Most isolates from Ragusano PDO cheese were identified as E. durans (46%) and/or E. faecalis (43%), while E. faecium (605) was mainly detected in Pecorino Siciliano cheese. High incidence of resistance (97% of total strains) was detected for rifampicin, erythromycin and ampicillin. Moreover, 83 isolates (75%) exhibited multidrug-resistant phenotypes and the one VRE (vanB) isolate was identified as E. durans. PFGE analysis clustered isolates into 22 genotypes and the presence of the same PFGE types, for both E. durans and E. faecalis , in the two cheese types, suggest the link between enterococci and geographical area of production. Results of present study raise concerns about possible role of dairy enterococci as reservoirs of antibiotic resistance. Highlights • E. durans and E. faecalis were found dominant in Ragusano PDO cheese. • E. faecium and E. faecalis were mainly isolated from Pecorino Siciliano cheese. • High incidence of resistance was detected for rifampicin, erythromycin and ampicillin. • 97% of strains exhibited multidrug-resistant phenotypes. • Same PFG enterococcal types, in both cheeses, suggest a link between enterococci and geographical area of production. [ABSTRACT FROM AUTHOR]

In the present study, two lactobacilli strains, Lactobacillus rhamnosus H25 and Lactobacillus paracasei N24, used as adjunct cultures, were evaluated for their heat resistance both with and without prior heat adaptation and for their survival, at industrial scale, during the production and ripening of the Pecorino Siciliano cheese. In addition, the viability and persistence of the lactobacilli strains after passage through the gastrointestinal tract of healthy volunteers were evaluated by using rep-PCR analysis of viable cells. Both strains exhibited good heat resistance and survival throughout cheese production and ripening, and positively influenced the physico-chemical, the microbiological and the sensorial characteristics of the final product. In addition, the molecular typing of the lactobacilli isolates, retrieved from fecal samples of healthy volunteers during and after 15 days of the experimental cheese administration, revealed a high survival of the strains, highlighting their persistence during passage into the GI tract. In conclusion, this study proposes the two adjunct cultures as potential probiotic candidate deliverable by cheese. [ABSTRACT FROM AUTHOR]

[Display omitted] • The membrane filtration allows to recovery biophenols from olive mill wastewater. • Olive mill wastewater is a source of biomolecules to obtain functional food. • An olive mill wastewater concentrate was added to a blood orange juice. • The fortified blood orange juice may be proposed as new functional food. In the present study, a tangential membrane filtration system was applied to recover phenols from olive mill wastewater. The obtained concentrates were characterised for physico-chemical traits, antioxidant activity and antimicrobial effects. Results indicated that the highest concentration of hydroxytyrosol (7203.7 mg/L) was detected in the concentrate obtained by reverse osmosis, which also showed the highest antioxidant and antimicrobial activity. Moreover, the same concentrate was added, at different ratio, up to 4:250 v/v, into a commercial blood orange juice. The fortified juice with the addition of the concentrate, up to 2:250 v/v ratio, did not show off-flavour and off-odour compared to the control. Furthermore, after 60 days of refrigerated storage, the fortified juice exhibited a hydroxytyrosol content still complying with the daily intake recommended by EFSA health claim. The obtained results can be industrially useful in producing orange juice added with a natural antioxidant concentrate as a 'clean label' ingredient. [ABSTRACT FROM AUTHOR]

• Streptococci and lactobacilli were among the major groups revealed by Illumina. • The inoculums added increased the biodiversity of the cheeses. • Fatty acid composition was not affected by the addition of LAB culture. • Control and experimental cheeses greatly differed for the content of butanoic acid. 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. [ABSTRACT FROM AUTHOR]