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This study aimed to determine the glycemic index (GI) of a commercial mixed fruit juice (apple, orange, grape, and pomegranate; FJ) fortified with vitamin D3 or n-3 polyunsaturated fatty acids (PUFA) or probiotics, and their combination, and their effects on glycemic responses and salivary insulin concentrations. In a randomized controlled, double-blind, crossover study, 11 healthy participants (25 ± 2 years; five women; body mass index = 23 ± 1 kg/m2) were randomly assigned to receive five types of FJs [vitD (with vitamin D3); n-3 (with n-3 PUFA); probiotics (with Lacticaseibacillus casei Shirota and Lacticaseibacillus rhamnosus GG); vitD-n-3-probiotics FJ (combination of vitD3-n-3-probiotics), control (regular FJ)], all containing 50 g available carbohydrate, and glucose as reference drink. All FJs provided low GI values (control: 54; vitD3: 52; n-3: 51; probiotics: 50; and vitD-n-3-probiotics combination: 52, on glucose scale). Compared to the FJ control, the enriched FJs produced different postprandial glycemic and insulinemic responses and affected satiety scores. All FJ types, regardless of the added biofunctional ingredients, attenuated postprandial glycemic responses, which may offer advantages to glycemic control.

Lactobacillus is a diverse genus that includes species of industrial and biomedical interest. Lactiplantibacillus pentosus, formerly known as Lactobacillus pentosus, is a recently reclassified species, that contains strains isolated from diverse environmental niches, ranging from fermented products to mammalian gut microbiota. Importantly, several L. pentosus strains present health-promoting properties, such as immunomodulatory and antiproliferative activities, and are regarded as potential probiotic strains. In this study, we present the draft genome sequence of the potential probiotic strain L. pentosus L33, originally isolated from fermented sausages. Comprehensive bioinformatic analysis and whole-genome annotation were performed to highlight the genetic loci involved in host-microbe interactions and the probiotic phenotype. Consequently, we found that this strain codes for bile salt hydrolases, adhesins and moonlighting proteins, and for Class IIb bacteriocin peptides lacking the GxxxG and GxxxG-like motifs, crucial for their inhibitory activity. Its adhesion ability was also validated in vitro, on human cancer cells. Furthermore, L. pentosus L33 contains an exopolysaccharide (EPS) biosynthesis cluster, and it does not carry transferable antibiotic resistance genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and CAZymes analyses showed that L. pentosus L33 possesses biosynthetic pathways for seven amino acids, while it can degrade a wide array of carbohydrates. In parallel, Clusters of Orthologous Groups (COGs) and KEGG profiles of L. pentosus L33 are similar to those of 26 L. pentosus strains, as well as of two well documented L. plantarum probiotic strains. Conclusively, L. pentosus L33 exhibits good probiotic potential, although further studies are needed to elucidate the extent of its biological properties.

The present study concerns the serious issue of biodeterioration of the caves belonging to natural and cultural heritage sites due to the development of various microorganisms. Thus, a series of 18 essential oils (EOs) extracted from various Greek plants were evaluated in vitro (concentrations of 0.1, 0.2, 0.5, 1.0 and 5.0% v/v) against 35 bacterial and 31 fungi isolates (isolated from a Greek cave) and the antimicrobial activity was evident through the changes in optical density of microbial suspensions. In continuance, eight (8) representative bacterial and fungal isolates were further used to evaluate the minimum inhibitory concentration (MIC) and non-inhibitory concentration (NIC) values of the most effective EOs. According to the results, two EOs of Origanum vulgare were the most effective by inhibiting the growth of all the tested microorganisms at 0.1% (v/v), followed by that of Satureja thymbra which inhibited all bacterial isolates at 0.1% (v/v) and fungal isolates at 0.1, 0.2 and 0.5% (v/v) (depending on the isolate). The MIC ranged between 0.015–0.157 and 0.013–0.156 (v/v) for the bacterial and fungal isolates respectively, depending on the case. The current study demonstrated that conventional biocides may be replaced by herbal biocides with significant prospects for commercial exploitation.

UHT bovine milk (low-fat) was inoculated with a starter culture and one adjunct strain, Lactobacillus plantarum 2035 or Lb. plantarum T571, with probiotic potential, and also with a cocktail mixture of three strains of Listeria monocytogenes at three different initial levels of inoculum and the milk was fermented to pH 4.5. Microbiological, physicochemical, molecular and sensory analyses were performed during storage at 4 and 12 °C. The probiotic samples displayed higher counts of lactic acid bacteria, higher acidity, lower pH and reduced counts of L. monocytogenes, in shorter time period than the control at 4 °C. Molecular analysis verified the presence of the probiotic strains until the end of storage at both temperatures and in adequate amounts; survival of L. monocytogenes strains depended on the treatment. Sensory evaluation showed that the probiotic fermented milks had desirable organoleptic characteristics, similar to the control.

Alicyclobacillus acidoterrestris is considered to be one of the most important target microorganisms in quality control of heat-processed acidic foods. This species can cause spoilage by forming spores with very high heat resistance; however, no gas production or visible changes occur in the contaminated product during storage. Thus, the differentiation of A. acidoterrestris from other Alicyclobacillus species is of great importance. Τhe present study aims to find a rapid method for the identification of A. acidoterrestris. To achieve this, 78 Alicyclobacillus isolates were subjected to PCR-RFLP (restriction fragment length polymorphism). Specifically, PCR products of amplified V1-V3 region of the 16S rRNA gene were digested with restriction endonuclease HhaI. According to the obtained results, all A. acidoterrestris isolates showed similar restriction fragments of the 16S rRNA gene and different from A. acidocaldarius and A. hesperidum. In conclusion, a single enzyme PCR-RFLP assay was developed and showed rapid, inexpensive and direct identification of Alicyclobacillus isolates. The application of this method will be useful to identify this contaminant in fruit juices.

The current study aimed to explore the performance of a probiotic Lactobacillus strain as an adjunct culture in yogurt production and to assess Fourier transform infrared spectroscopy as a rapid, noninvasive analytical technique to evaluate the quality and the shelf life of yogurt during storage. In this respect, bovine milk (full-fat) was inoculated with the typical yogurt starter culture without (control case) or with the further addition of Lactobacillus plantarum T571 as an adjunct (probiotic case). The milk was also inoculated with a cocktail mixture of three strains of Listeria monocytogenes in two different initial levels of inoculum, and the fermentation process was followed. Accordingly, yogurt samples were stored at 4 and 12°C, and microbiological, physicochemical, molecular, and sensory analyses were performed during storage. Results showed that the lactic acid bacteria exceeded 7 log CFU/g during storage in all samples, where the probiotic samples displayed higher acidity, lower pH, and reduced counts of Lb. monocytogenes in a shorter period than the control ones at both temperatures. Pulsed-field gel electrophoresis verified the presence of the probiotic strain until the end of storage at both temperatures and in adequate amounts, whereas the survival and the distribution of Listeria strains depended on the case. The sensory evaluation showed that the probiotic samples had desirable organoleptic characteristics, similar to the control. Finally, the spectral data collected from the yogurt samples during storage were correlated with microbiological counts and sensory data. Partial least squares and support vector machine regression and classification models were developed to provide quantitative estimations of yogurt microbiological counts and qualitative estimations of their sensory status, respectively, based on Fourier transform infrared fingerprints. The developed models exhibited satisfactory performance, and the acquired results were promising for the rapid estimation of the microbiological counts and sensory status of yogurt.

Chicken is one of the most widely consumed meats worldwide. The exploration of the bacterial diversity of chicken meat may provide new insights into the chicken-associated microbiome that will lead to moderation of food spoilage or safety. This study was undertaken to explore the bacterial communities of chicken breast and thigh fillets stored at refrigeration (0 °C and 5 °C) and slightly abuse (10 °C) temperatures for 5 days through conventional cultural methods along with next-generation sequencing (NGS) analysis. Total viable counts (TVC), Brochothrix thermosphacta, Pseudomonas spp., and lactic acid bacteria (LAB) were enumerated, while the bacterial communities were mapped through 16S rRNA gene amplicon sequencing. Chicken breast and thigh fillets possessed a complex bacterial structure that incorporated a total of &gt, 200 Operational Taxonomic Units (OTUs) at the genus level. The core microbiota of fresh samples consisted of Acinetobacter, Brochothrix, Flavobacterium, Pseudomonas, Psychrobacter, and Vibrionaceae (family). These genera persisted until the end of storage in &gt, 80% of samples, except Psychrobacter and Flavobacterium, while Photobacterium was also identified. Hierarchical clustering showed a distinction of samples based on storage time and chicken part. Conventional plate counting with growth media commonly used in spoilage studies did not always correspond to the microbial community profiles derived from NGS analysis, especially in Pseudomonas, Acinetobacter, Photobacterium, and Vibrionaceae. Results of the present study highlight Photobacterium and Vibrionaceae, in general, as potent chicken meat spoilers and suggest the necessity to combine classical microbiological methods along with NGS technologies to characterize chicken meat spoilage microbiota.

Anigrides Nymphes of Lake Kaiafas is a thermal spring that is well known for its therapeutical properties, as the hot water (32&ndash, 34 &deg, C) is rich in sulfur compounds and minerals. Nowadays, efforts are made from the Hellenic Republic to modernize the existing facilities and infrastructure networks of the area. To study the complex ecosystem of the thermal spring, we collected water from four sampling points (Lake, and Caves 1, 2, and 3). Filtration method was used for microbial enumeration. In parallel, total bacterial DNA was extracted and subjected to next-generation sequencing (NGS). A total of 166 different bacterial families were detected. Differences in families, genera, and species abundances were detected between the different sampling points. Specifically, Comamonadaceae was the most common family detected in Lake and Cave 3. Similarly, in Caves 1 and 2, Rhodobacteraceae was detected at a higher percentage compared to the rest of the families. Moreover, the detection of sequences assigned to waterborne or opportunistic pathogens, i.e., Enterobacteriaceae, Legionellaceae, Coxiellaceae, and Clostridiaceae, as well as Enterococcus and Vibrio, is of great importance. Although the presence of pathogens was not examined by quantitative PCR, the detection of their sequences strengthens the need of the planned rehabilitation actions of this natural environment in order to allow human swimming.

Chicken liver is a highly perishable meat product with a relatively short shelf-life and that can get easily contaminated with pathogenic microorganisms. This study was conducted to evaluate the behavior of spoilage microbiota and of inoculated Salmonella enterica on chicken liver. The feasibility of Fourier-transform infrared spectroscopy (FTIR) to assess chicken liver microbiological quality through the development of a machine learning workflow was also explored. Chicken liver samples [non-inoculated and inoculated with a four-strain cocktail of ca. 103 colony-forming units (CFU)/g Salmonella] were stored aerobically under isothermal (0, 4, and 8°C) and dynamic temperature conditions. The samples were subjected to microbiological analysis with concomitant FTIR measurements. The developed FTIR spectral analysis workflow for the quantitative estimation of the different spoilage microbial groups consisted of robust data normalization, feature selection based on extra-trees algorithm and support vector machine (SVM) regression analysis. The performance of the developed models was evaluated in terms of the root mean square error (RMSE), the square of the correlation coefficient (R2), and the bias (Bf) and accuracy (Af) factors. Spoilage was mainly driven by Pseudomonas spp., followed closely by Brochothrix thermosphacta, while lactic acid bacteria (LAB), Enterobacteriaceae, and yeast/molds remained at lower levels. Salmonella managed to survive at 0°C and dynamic conditions and increased by ca. 1.4 and 1.9 log CFU/g at 4 and 8°C, respectively, at the end of storage. The proposed models exhibited Af and Bf between observed and predicted counts within the range of 1.071 to 1.145 and 0.995 to 1.029, respectively, while the R2 and RMSE values ranged from 0.708 to 0.828 and 0.664 to 0.949 log CFU/g, respectively, depending on the microorganism and chicken liver samples. Overall, the results highlighted the ability of Salmonella not only to survive but also to grow at refrigeration temperatures and demonstrated the significant potential of FTIR technology in tandem with the proposed spectral analysis workflow for the estimation of total viable count, Pseudomonas spp., B. thermosphacta, LAB, Enterobacteriaceae, and Salmonella on chicken liver.

In this work, the inactivation kinetics of Alicyclobacillus acidoterrestris spores by temperature-assisted high hydrostatic pressure was assessed by means of the Weibull model. Spores from two A. acidoterrestris strains (a wild-type strain and a reference strain) were inoculated in commercial orange juice and subjected to high pressure levels (500 and 600 MPa) combined with four temperature regimes (25, 45, 60 and 70 &deg, C) for time up to 30 min. Results showed that for a given high-pressure level spore inactivation was higher as temperature progressively increased. Furthermore, the Weibull model consistently produced satisfactory fit to the inactivation data based on the values of the root mean squared error (RMSE &lt, 0.54 log colony-forming units (CFU)/mL) and the coefficient of determination (R2 &gt, 0.90 in most cases). The shape of inactivation curves was concave upward (p &lt, 1) for all temperature/high pressure levels tested, indicating rapid inactivation of the sensitive cells of the bacterium whereas the remaining ones adapted to high hydrostatic pressure (HHP) treatment. The values of the shape (p) and scale (&delta, ) parameters of the Weibull model were dependent on the applied temperature for a given high pressure level and they were further described in a secondary model using first-order fitting curves to provide predictions of the surviving spore population at 55 and 65 &deg, C. Results revealed a systematic over-prediction for the wild-type strain regardless of temperature and high pressure applied, whereas for the reference strain under-prediction was evident after 3 log-cycles reduction of the surviving bacteria spores. Overall, the results obtained indicate that the effectiveness of high hydrostatic pressure against A. acidoterrestris spores is strain-dependent and also underline the need for temperature-assisted HPP for effective spore inactivation during orange juice processing.

The aim of the present study was to investigate the evolution of the volatile compounds of aerobically stored sterile pork meat as a consequence of the metabolic activities of inoculated specific spoilage microorganisms. Thus, Pseudomonas fragi, Pseudomonas putida, Lactobacillus sakei and Leuconostoc mesenteroides were inoculated in monocultures, dual cultures and a cocktail culture of all strains on sterile pork meat stored aerobically at 4 and 10 &deg, C. Microbiological and sensory analyses, as well as pH measurements, were performed, along with headspace solid-phase microextraction gas chromatography/mass spectroscopy (headspace SPME&ndash, GC/MS) analysis. Data analytics were used to correlate the volatile compounds with the spoilage potential of each stain using multivariate data analysis. The results for the sensory discrimination showed that the volatiles that dominated in spoiled samples consisted mostly of alcohols, ketones and two esters (butyl acetate and ethyl acetate), while at fresh samples, dimethyl sulfide, furans, acetoin and ethyl lactate were detected. On the other hand, 2-butanone, diacetyl and acetaldehyde were among the volatile compounds that were mainly correlated with the inoculated meat during storage. In addition, P. fragi was positively correlated with a higher number of volatiles compared to the other strains, strengthening the hypothesis that volatile compound production is strain-dependent.

Autochthonous Saccharomyces cerevisiae vineyard populations are important components of the grape/wine system. Besides their direct impact on winemaking, they also constitute an untapped reservoir of genotypes with special technological attributes for the wine industry. Research so far on S. cerevisiae populations has focused on spatial distribution on large scales, yet little is known about the genetic variability of populations within viticultural zones and their temporal genotypic variation. Here, S. cerevisiae populations from different vineyards in Santorini, a small Aegean island, were genotyped and their genetic diversity was assessed within and between vineyards during two consecutive years. Despite the relative geographical isolation of the island, a relatively high genetic diversity was uncovered. The vast majority of genotypes were vineyard-specific, while in one of the vintages, significant differences in the genotypic composition of vineyards were detected. Overall, higher differences were detected between vintages rather than among vineyards. Notably, only four genotypes were common for the two vintages, three of which were commercial S. cerevisiae strains, probably &ldquo, escapees&rdquo, from wineries. Nevertheless, the populations of the two vintages were not genetically distinct. Present results highlight the magnitude of genetic diversity in natural wine yeast populations on a small spatial scale, yet the invasion of commercial starters may constitute a potential risk for loss of local yeast biodiversity. However, present results show that industrial strains do not necessarily dominate over the natural strains or their high abundance may be temporary.

The aim of the present study was to investigate the evolution of the volatile compounds of aerobically stored sterile pork meat as a consequence of the metabolic activities of inoculated specific spoilage microorganisms. Thus, Pseudomonas fragi, Pseudomonas putida, Lactobacillus sakei and Leuconostoc mesenteroides were inoculated in monocultures, dual cultures and a cocktail culture of all strains on sterile pork meat stored aerobically at 4 and 10 °C. Microbiological and sensory analyses, as well as pH measurements, were performed, along with headspace solid-phase microextraction gas chromatography/mass spectroscopy (headspace SPME–GC/MS) analysis. Data analytics were used to correlate the volatile compounds with the spoilage potential of each stain using multivariate data analysis. The results for the sensory discrimination showed that the volatiles that dominated in spoiled samples consisted mostly of alcohols, ketones and two esters (butyl acetate and ethyl acetate), while at fresh samples, dimethyl sulfide, furans, acetoin and ethyl lactate were detected. On the other hand, 2-butanone, diacetyl and acetaldehyde were among the volatile compounds that were mainly correlated with the inoculated meat during storage. In addition, P. fragi was positively correlated with a higher number of volatiles compared to the other strains, strengthening the hypothesis that volatile compound production is strain-dependent.

The aim of the present study was to assess the influence of ripening container’s material on the bacterial diversity of Feta cheese PDO (Protected Designation of Origin). The microbiota of fresh and mature cheese produced in plastic and stainless steel container was monitored by microbial enumeration and 16s rRNA gene sequencing. According to the obtained results, lactic acid bacteria (LAB) was the dominant microbiota of fresh and mature cheese. Metagenomics data revealed that fresh cheese was dominated by Lactococcus followed by members of Enterobacteriaceae family and Pseudomonas. Similarly, Lactococcus was the most abundant genus detected in mature cheese (54 days and 120 days), regardless of the container’s material. In both fresh and mature cheese, species of Pseudomonas, Streptococcus, Acinetobacter, Lactobacillus, Flavobacterium, and Carnobacterium were detected. The abundance of Enterobacteriaceae, Moraxellaceae and Pseudomonadaceae in mature cheese ripened in stainless steel container seems to be numerically reduced after 120 days of storage compared to the cheese ripened in plastic container but not significant differences were observed (p > 0.05). In conclusion, metagenomic analysis suggests that ripening container’s material does not affect the microbial community responsible for the ripening of feta cheese PDO.

Survival during transit through the gastrointestinal track, intestinal mucosa adhesion, and a potential immunomodulatory effect of Lactobacillus plantarum strains 2035 and ACA-DC 2640 were investigated in a rat model. According to microbiological and multiplex PCR analysis, both strains were detected in feces 24 h after either single-dose or daily administration for 7 days. Intestinal mucosa adhesion of L. plantarum 2035 was noted in the large intestine at 24 h after single-dose administration, while it was not detected at 48 h. Daily dosing, prolonged detection of the strain up to 48 h post-administration, and expanded adhesion to the small intestine. Adhesion of L. plantarum ACA-DC 2640 to the intestinal mucosa after single-dose administration was prolonged and more extended compared to L. plantarum 2035. Daily dosing increased both the levels and the rate of positive cultures of the strains compared to those of the single-dose scheme. In addition, both strains increased total IgG while decreased IgM and IgA serum levels. In conclusion, L. plantarum 2035 and L. plantarum ACA-DC 2640 survived transit through the gastrointestinal track, exhibited transient distinct adhesion to the intestinal mucosa and modulated the systemic immune response.

The aim of the present study was to assess application of free or immobilized kefir culture on apple pieces and delignified cellulosic material (DCM) in simultaneous alcoholic and malolactic cider fermentations at a wide temperature range (5–45 °C). Repeated batch fermentations were continued for higher than 7 months, showing a high operational stability of the systems and were completed in less than 24 h with immobilized cells on DCM at 37 °C. Malic acid conversion up to 71.5% and ethanol productivity values up to 56.9 g/(Ld) were recorded, which could be adopted by the industrial sector. PCR-DGGE analysis of kefir culture showed no changes in microbial diversity during cell immobilization and fermentations. Analysis of major volatiles documented a significant increase of ethyl acetate content and a decrease of higher alcohols at low temperatures. HS-SPME GC/MS analysis revealed that the highest content of esters and total volatiles was observed in cider fermented by kefir culture immobilized on apple pieces at 20 °C. Principal Component Analysis showed that mainly the immobilization support rather than the temperature had a significant effect on volatile composition. Finally, the sensory evaluation ascertained the high quality of the new products.

Τhe aim of the present study was to evaluate the production of probiotic yoghurts using immobilized cells of Lactobacillus plantarum 2035 on whey protein along with a commercial culture. For comparison reasons, yoghurts with free L. plantarum 2035 cells and with no probiotic culture were also prepared. All products using either bovine or ewe's milk were prepared by our industrial partner (RODOPI SA dairy industry). The pH ranged between 4.1–4.4 in all cases, while the rest physicochemical characteristics ranged within usual levels. Microbiological and strain specific multiplex PCR analysis confirmed that both free and immobilized L. plantarum 2035 cells were detected in yoghurts at levels ≥6 log cfu/g after storage at 4 °C for longer period than the expiration day (≥4 weeks). Principal Component Analysis of the compounds identified by HS-SPME GC/MS analysis revealed that the milk origin affected mainly the volatile composition. Finally, the organoleptic assessment demonstrated that the new yoghurts exhibited improved sensorial characteristics compared to the commercial products.

The aim of the study was to evaluate the efficacy of oregano essential oil (OEO) incorporated in Na-alginate edible films when applied to sliced ham inoculated with a cocktail of Listeria monocytogenes strains, with or without pretreatment by high pressure processing (HPP). Microbiological, physicochemical and sensory analyses (in Listeria-free slices) were performed, while, the presence/absence and the relative abundance of each Listeria strain, was monitored by pulsed field gel electrophoresis (PFGE). The OEO incorporation in the films, caused approximately 1.5 log reduction in Listeria population at 8 and 12 &deg, C at the end of the storage period, and almost 2.5 log reduction at 4 &deg, C. The HPP treatment caused 1 log reduction to the initial Listeria population, while levels kept on decreasing throughout the storage for all the tested temperatures. The pH of the samples was higher in the cases where HPP was involved, and the samples were evaluated as less spoiled. Furthermore, the presence of OEO in the films resulted in color differences compared to the control samples, whilst the aroma of these samples was improved. In conclusion, the combined application of HPP and OEO edible films on the slices, led to a significant reduction or absence of the pathogen.

Multi-starter wine fermentations employing non-Saccharomyces (NS) yeasts are becoming an emerging trend in winemaking. It is therefore important to determine the impacts of different NS strains in the wine phenotype and in particular the aroma outputs in different inoculation schemes and fermentation conditions. Here, two native NS yeasts, Lachancea thermotolerans LtMM7 and Hanseniaspora uvarum HuMM19, were assessed for their ability to improve the quality of Moschofilero, a Greek aromatic white wine. The NS strains were initially examined in laboratory scale fermentations in mixed inoculations with ScMM23, a native Saccharomyces cerevisiae strain. LtMM7 was selected to be further evaluated in pilot scale fermentations. Five different inoculation schemes were considered: single inoculation of ScMM23 (IS), simultaneous inoculation of ScMM23 with HuMM19 (SMH) or LtMM7 (SML), and sequential inoculation of HuMM19 (SQH) or LtMM7 (SQL) followed by ScMM23. At laboratory scale fermentations, the chemical profiles were largely affected by both the NS species and the inoculation scheme applied. The sequential inoculation using HuMM19 produced the most divergent wine phenotype. However, HuMM19 caused significant increases in acetic acid and ethyl acetate levels that impeded its use in pilot scale trials. LtMM7 significantly affected the chemical profiles of wines produced at the winery, especially in the sequential inoculation scheme. Importantly, LtMM7 significantly increased the levels of acetate esters or ethyl esters, depending on the inoculation method applied. In particular, acetate esters like isobutyl acetate, hexyl acetate, and 2-phenylethyl acetate, which all impart fruity or floral aromas, were significantly increased in SQL. On the other hand, higher levels of total ethyl esters were associated with SML. The most striking differences were observed in the levels of fruit-impair esters like ethyl decanoate, 3-methylbutyl octanoate, and isoamyl hexanoate. This is the first study to report a significant increase in the ethyl ester fraction by L. thermotolerans. Interestingly, L. thermotolerans in SQL also increased the concentrations of damascenone and geraniol, the major teprenic compound of Moschofilero, which are associated with several typical floral and fruity aromas of the variety. Present results show that L. thermotolerans may enhance the varietal character and increase the chemical complexity of Moschofilero wines.

The beneficial effects of two potential probiotic lactobacillus strains, namely Lactobacillus pentosus B281 and L. plantarum B282 were examined and the mechanisms of action were investigated. Quantitative analysis and confocal microscopy showed that both strains exhibited a significant higher adherence to Caco-2 cells in comparison to the reference strain L. casei ATCC 393. Treatment with conditioned media (CM) of the two strains caused significant reduction of cell proliferation, as demonstrated by Sulforhodamine B (SRB) and clonogenic assays. Moreover, the CM of the two strains induced a G1-phase arrest and down-regulation of specific cyclin genes, as indicated by flow cytometry and real-time PCR analysis. To begin elucidating the nature of the bacterial components conveying these responses, the anti-proliferative effect of heat-treated CM was analysed. The anti-proliferative activity of heat-treated CM was similar to the non-heated CM in a time- and dose-dependent manner, indicating the presence of thermostable bioactive compounds.

Current information from conventional microbiological methods on the microbial diversity of table olives is insufficient. Next-generation sequencing (NGS) technologies allow comprehensive analysis of their microbial community, providing microbial identity of table olive varieties and their designation of origin. The purpose of this study was to evaluate the bacterial and yeast diversity of fermented olives of two main Greek varieties collected from different regions&mdash, green olives, cv. Halkidiki, from Kavala and Halkidiki and black olives, cv. Konservolia, from Magnesia and Fthiotida&mdash, via conventional microbiological methods and NGS. Total viable counts (TVC), lactic acid bacteria (LAB), yeast and molds, and Enterobacteriaceae were enumerated. Microbial genomic DNA was directly extracted from the olives&rsquo, surface and subjected to NGS for the identification of bacteria and yeast communities. Lactobacillaceae was the most abundant family in all samples. In relation to yeast diversity, Phaffomycetaceae was the most abundant yeast family in Konservolia olives from the Magnesia region, while Pichiaceae dominated the yeast microbiota in Konservolia olives from Fthiotida and in Halkidiki olives from both regions. Further analysis of the data employing multivariate analysis allowed for the first time the discrimination of cv. Konservolia and cv. Halkidiki table olives according to their geographical origin.

The performance of the Lactobacillus plantarum L125 strain with probiotic potential, previously isolated from a traditional Greek fermented sausage, was assessed as an adjunct culture for the production of dry-fermented pork sausages. The probiotic was added in the meat batter with the commercial starter cultures and the microbial populations were estimated during fermentation, ripening and storage at 4 and 12 °C, respectively. Physicochemical analyses (pH and aw), sensory evaluation and Fourier Transform Infrared Spectroscopy (FTIR) were also performed, while the presence of L125 strain was monitored by Pulsed Field Gel Electrophoresis. Lactic acid bacteria (LAB) counts at the inoculation time were 8.21 and 6. 44 log CFU/g for the probiotic and control samples, respectively, while the L125 strain maintained high population levels (> 6 log CFU/g) during storage. At the end of the ripening, the products exhibited similar pH values of 4.46 and 4.42 and aw values of 0.87 and 0.85 for the control and probiotic case, respectively. FTIR spectroscopy application in combination with data analytics, resulted in the acceptable estimation of LAB population, but further studies are needed to improve the technique's performance. Overall, the Lb. plantarum L125 was found to possess desirable technological characteristics, indicating its effectiveness to be used in fermented sausage manufacturing.

Green olives cv. Halkidiki previously fermented by the potential probiotic Lactobacillus pentosus B281, Lactobacillus plantarum B282, or the indigenous microbiota were packed under modified atmospheres (70% N2–30% CO2) and stored at 4 and 20 °C for 12 months. The populations of lactic acid bacteria (LAB) and yeasts as well as the physicochemical characteristics of olives (pH, water activity, colour, acidity, salt content and firmness) were monitored. The LAB population was 5.6–5.9 at the beginning of the storage and reached 5.8–6.4 log CFU/g after one year. Pulsed-field gel electrophoresis revealed survival rates of 100% for L. pentosus B281 and 46.7% for L. plantarum B282 at the beginning of storage. Both strains were recovered in high percentages (64.7% for B281 and 94.1% for B282) after 6 months and in lower survival rates after one year (35.0% for B281 and 13.3% for B282) at 4 °C. Lower rates were also observed during storage at 20 °C. Moreover, the product was organoleptically more acceptable after 6 than 12 months of storage at both temperatures. Thus, a shelf life of six months at refrigerating temperature could be proposed as the best studied condition to deliver a promising probiotic food to the consumer.

Nowadays, the consumption of food products containing probiotics, has increased worldwide due to concerns regarding healthy diet and wellbeing. This trend has received a lot of attention from the food industries, aiming to produce novel probiotic foods, and from researchers, to improve the existing methodologies for probiotic delivery or to develop and investigate new possible applications. In this sense, edible films and coatings are being studied as probiotic carriers with many applications. There is a wide variety of materials with film-forming ability, possessing different characteristics and subsequently affecting the final product. This manuscript aims to provide significant information regarding probiotics and active/bioactive packaging, to review applications of probiotic edible films and coatings, and to discuss certain limitations of their use as well as the current legislation and future trends.

The aim of the study was to investigate the performance of Lactobacillus plantarum T571 with probiotic potential as a co-starter culture in Feta cheese production and in its long-term storage. For this reason, Feta cheese was manufactured without (control) or with the probiotic T571 strain (probiotic) and then monitored during storage at 4 and 12 °C, respectively. The products were also inoculated with Listeria monocytogenes (3-strain cocktail). The results showed that lactic acid bacteria exceeded 6 log CFU/g during storage in all trials. The probiotic samples displayed higher acidity (≈1.5% lactic acid) and lower pH (≈4.2). Coliforms and L. monocytogenes, were inactivated in shorter time in probiotic samples, in comparison with the control ones. Pulsed field gel electrophoresis verified the presence of the probiotic strain in the cheese, until the end of storage at both temperatures, whilst the survival and distribution of the pathogenic strains depended on the trial. The sensory evaluation showed that the probiotic cheeses had desirable sensory characteristics similar to the control ones, with scores of ca. 8 on a 10 cm-scale. Finally, Fourier transform infrared spectroscopy was applied for the first time during Feta cheese storage with promising results for the rapid estimation of the microbiological counts and sensory status of cheese. Results showed that Lb. plantarum T571 highlighted desirable and robust technological properties and may be used in cheese making as an adjunct culture.

The aim of the present study was to investigate the potential use of Fourier transform infrared (FTIR) spectroscopy to quantify biochemical changes occurring in ham slices packed with probiotic supplemented edible films and treated with High Pressure Processing (HPP), in monitoring spoilage. Details regarding the data collection and experimental procedure were presented by Pavli et al. (2017). A series of Partial Least Squares (PLS) models were developed to correlate spectral data from FTIR analysis with ham spoilage during storage under vacuum at different temperatures (4, 8 and 12°C). FTIR spectra were collected from the surface of the ham samples in parallel with microbiological analysis of total viable counts (TVC) and lactic acid bacteria (LAB). Qualitative interpretation of spectral data was based on a sensory evaluation, using a hedonic scale, classifying the samples in three quality classes, fresh, semi-fresh and spoiled. The scope of the modeling approach was to discriminate the ham slices in their respective quality class and additionally to predict the microbial population directly from spectral data. The results obtained demonstrated that the processing of the samples affected the performance of classification in the sensory classes, with better results observed in the case of for ham slices packed with probiotic supplemented (PS) edible films and of control samples without HPP. The performance of PLS regression models on providing quantitative estimations of microbial counts were based on specific figures of merit (bias factor, accuracy factor, root mean square error, percentage of prediction error). Bias and accuracy factors were close to unity for both microbial groups tested for samples without HPP, whereas for HPP treated samples the values of these indices ranged from 0.963 to 1.332, depending on the case and indice. The results of this study demonstrated for the first time that although FTIR can be used reliably for the rapid assessment of sliced ham, additional processes such as HPP can affect its performance.

The performance of two strains of lactic acid bacteria (LAB), namely Lactobacillus pentosus B281 and Lactobacillus plantarum B282, previously isolated from industrially fermented table olives and screened in vitro for probiotic potential, was investigated as starter cultures in Spanish style fermentation of cv. Halkidiki green olives. Fermentation was undertaken at room temperature in two different initial salt concentrations (8% and 10%, w/v, NaCl) in the brines. The strains were inoculated as single and combined cultures and the dynamics of their population on the surface of olives was monitored for a period of 114 days. The survival of inoculated strains on olives was determined using Pulsed Field Gel Electrophoresis (PFGE). Both probiotic strains successfully colonized the olive surface at populations ranged from 6.0 to 7.0 log CFU/g throughout fermentation. PFGE analysis revealed that L. pentosus B281 presented higher colonization in both salt levels at the end of fermentation (81.2% and 93.3% in 8% and 10% NaCl brines, respectively). For L. plantarum B282 a high survival rate (83.3%) was observed in 8% NaCl brines, but in 10% NaCl the strain could not colonize the surface of olives. L. pentosus B281 also dominated over L. plantarum B282 in inoculated fermentations when the two strains were used as combined culture. The biochemical profile (pH, organic acids, volatile compounds) attained during fermentation and the sensory analysis of the final product indicated a typical lactic acid fermentation process of green olives.