Preserving the efficacy of probiotic bacteria exhibits paramount challenges that need to be addressed during the development of functional food products. Several factors have been claimed to be ...responsible for reducing the viability of probiotics including matrix acidity, level of oxygen in products, presence of other lactic acid bacteria, and sensitivity to metabolites produced by other competing bacteria. Several approaches are undertaken to improve and sustain microbial cell viability, like strain selection, immobilization technologies, synbiotics development etc. Among them, cell immobilization in various carriers, including composite carrier matrix systems has recently attracted interest targeting to protect probiotics from different types of environmental stress (e.g., pH and heat treatments). Likewise, to successfully deliver the probiotics in the large intestine, cells must survive food processing and storage, and withstand the stress conditions encountered in the upper gastrointestinal tract. Hence, the appropriate selection of probiotics and their effective delivery remains a technological challenge with special focus on sustaining the viability of the probiotic culture in the formulated product. Development of synbiotic combinations exhibits another approach of functional food to stimulate the growth of probiotics. The aim of the current review is to summarize the strategies and the novel techniques adopted to enhance the viability of probiotics.
Cheese whey constitutes one of the most polluting by-products of the food industry, due to its high organic load. Thus, in order to mitigate the environmental concerns, a large number of valorization ...approaches have been reported; mainly targeting the recovery of whey proteins and whey lactose from cheese whey for further exploitation as renewable resources. Most studies are predominantly focused on the separate implementation, either of whey protein or lactose, to configure processes that will formulate value-added products. Likewise, approaches for cheese whey valorization, so far, do not exploit the full potential of cheese whey, particularly with respect to food applications. Nonetheless, within the concept of integrated biorefinery design and the transition to circular economy, it is imperative to develop consolidated bioprocesses that will foster a holistic exploitation of cheese whey. Therefore, the aim of this article is to elaborate on the recent advances regarding the conversion of whey to high value-added products, focusing on food applications. Moreover, novel integrated biorefining concepts are proposed, to inaugurate the complete exploitation of cheese whey to formulate novel products with diversified end applications. Within the context of circular economy, it is envisaged that high value-added products will be reintroduced in the food supply chain, thereby enhancing sustainability and creating "zero waste" processes.
The in vitro and in situ antifungal activity of Lactobacillus plantarum against the ochratoxigenic fungus Aspergillus carbonarius was investigated in this study. Four different fungal isolates from ...grape berries were co-cultured with four different strains of L. plantarum on Malt Extract Agar (MEA) plates at 30 °C. Bacterial strains inhibited fungal growth up to 88% and significantly reduced toxin production up to 100%. In addition, L. plantarum was evaluated as biocontrol agent against A. carbonarius growth and OTA production on table grapes. Temporal studies of bacterial antagonism were performed with two different grape cultivars. Artificially wounded and unwounded berries were pre-treated with 108 CFU/mL bacteria and inoculated with 106 spores/mL of A. carbonarius ochratoxigenic isolates. Biocontrol agents displayed high rate of colonization on grapes during 5 days of incubation at 30 °C. Scanning electron microscopy (SEM) also determined the presence of microorganisms on grape surface. Bacterial strains were effective in controlling fungal infection reaching up to 71% inhibition rates. However the presence of wounds on grape skin facilitated infection of berries by A. carbonarius, since unwounded berries showed lower levels of infection. Results also revealed significant reduction in mycotoxin production ranging between 32% and 92%. Transcriptome analysis following exposure to co-cultivation, exhibited differential expression for each gene studied of AcOTAnrps (Aspergillus carbonarius OTA nonribosomal), AcOTApks (Aspergillus carbonarius OTA polyketide synthase) and laeA, emphasizing the significance of strain variability. The genes AcOTAnrps and laeA were most influenced by the presence of L. plantarum. This work is a contribution for the potential biocontrol of toxigenic fungi in table grapes by lactic acid bacteria (LAB). The above findings underline the significance of bacterial strain variability on the effectiveness of biopreservative features of L. plantarum strains.
•L. plantarum presented strain dependent antifungal effect on A. carbonarius growth.•Bacteria successfully colonized and influenced fungal toxin production on grapes.•Intra-species differences on gene expression were revealed for A. carbonarius isolates.•Genes OTAnrps and laeA were significantly affected by bacterial co-culture.
Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS), a culture based alternative for microbial diversity studies, is an attractive tool to dereplicate large ...numbers of isolates to a smaller set of representatives for downstream characterization. In the present study, MALDI-TOF MS, combined with a database of reference spectra compiled in previous studies, was applied to identify 88 non-starter lactic acid bacteria (NSLAB) isolated from 18 samples of four different artisanal cheeses produced in the Island of Naxos, Greece, from raw sheep and goat milk without the addition of starters. Eighty-four isolates (95.5%) could be identified directly via MALDI-TOF MS. Lactobacillus brevis and Lactobacillus plantarum were the dominant species, followed by Lactococcus lactis, Leuconostoc mesenteroides Lactobacillus paracasei, Lactobacillus rhamnosus, Pediococcus pentosaceus and Enterococcus faecium. The remaining four isolates represented species present in the database; however, within-species diversity was insufficiently covered. Additionally, pheS sequencing was applied to confirm identification.
•Application of MALDI-TOF MS for the identification of lactic acid bacteria•First fingerprinting of the non-starter lactic acid bacteria of artisanal cheeses produced in the Island of Naxos, Greece•PheS sequencing was performed to validate MALDI-TOF MS identication
The objective of this study was to investigate the effect of environmental factors, namely temperature and water activity, on genes involved in the regulation of ochratoxin A (OTA) production over ...time. For this purpose, the previously characterized toxigenic
Ac29 isolate from Greek vineyards and the
ITEM 5010 reference strain were subjected to combined temperature and water activity (a
) treatments to study OTA production and relative gene expression. The fungal isolates were grown on a synthetic grape juice liquid medium (SGM) under different temperature (20 °C, 25 °C and 30 °C) and a
(0.94 and 0.98) regimes. The expression of the AcOTA
, AcOTA
, and
OTA related genes was investigated using real time PCR. Gene expression was monitored at the same time points, along with fungal biomass and OTA accumulation at three, six and nine days of incubation. In gene expression analysis, stimulation of the biosynthetic genes was observed a few days before any toxin could be detected. This fact may underline a possible early indicator of potential toxin contamination of grapes. However, the transcript levels varied with respect to the different combinations of ecophysiological conditions and time, highlighting a complex regulation of OTA related gene expression of
in the specific medium.
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• Indigenous yeast strains contribute to greater expression of terroir fingerprint.• High-throughput sequencing confers an emerging tool for novel yeast strains identification.• ...Multi-starter cultures fermentation using non-conventional strains is the new trend in modern vinification.• Interspecies hybridization techniques may generate Saccharomyces phenotypes of special oenological traits.
Research on grape microbiome dynamics inaugurates new perspectives for sophisticated complex bio-transformations that can profoundly enhance wine character. The exploitation of the significant variations in yeast biota of grapevines in different viticultural areas entails the potential to confer specific regional character in wines. Thereby, numerous studies are following this flourishing oenological interest on distinct microbial fingerprints and dynamic interactions over fermentation. Premium quality ‘natural’ wines are an emerging topic of the current decade in winemaking. Thus, research is directed towards the characterization of genotypes of indigenous yeasts and their beneficial role in unique wine profiles. The identification of new fermentative yeast strains undertakes continuous challenges in the production of top-of-the-line wines with exclusive regional traits. Herein, distinctive major trends in winemaking are reviewed whereas novel and promising studies are being highlighted.
The development of innovative functional products with potential health benefits, under the concept of bio-economy, is flourishing. This study undertook an evaluation of non-dairy lactobacilli ...Lactiplantibacillus pentosus B329 and Lactiplantibacillus plantarum 820 as “ready to use” starter cultures. Lactic acid bacteria (LAB) cultures were evaluated for their fermentation efficiency, before and after freeze-drying, using cheese whey (CW) as a fermentation substrate and subsequent immobilization on bacteria cellulose (BC) to produce a novel biocatalyst. The biocatalyst was applied in functional sour milk production and compared with free cells via the assessment of physicochemical and microbiological properties and sensory evaluation. Evidently, LAB strains exhibited high fermentative activity before and after freeze-drying. Results of a 5-month storage stability test showed that viability was 19% enhanced by immobilization on BC, supporting the concept of “ready to use” cultures for the production of fermented beverages. Likewise, sour milk produced by the BC biocatalyst presented higher organoleptic scores, compared to the free cells case, whereas immobilization on BC enhanced probiotic viability during post-fermentation storage (4 °C, 28 days). The obtained high viability (>107 log cfu/g) demonstrated the efficacy of the proposed bioprocess for the production of functional/probiotic-rich beverages. Ultimately, this work presents a consolidated scheme that includes the advantages and the cooperative effect of probiotic LAB strains combined with a functional biopolymer (BC) towards the formulation of novel functional products that coincide with the pillars of food systems sustainability.
The in vitro susceptibility to sodium metabisulphite (NaMBS) was investigated in 10 different food spoilage filamentous fungi, namely Aspergillus flavus, A. carbonarius, A. niger, A. ochraceus, A. ...tubingensis, A. westerdijkiae, Cladosporium cladosporioides, Fusarium oxysporum, Penicillium commune and P. expansum. The fungi were inoculated in sterile 96-well microtiter plates containing Yeast-extract Sucrose (YES) semi-solid agar supplemented with NaMBS in concentrations ranging from 2000 to 3.9 mg l−1 and incubated at 25 °C. Growth was monitored by absorbance measurements at 600 nm using a multi-spectrophotometer. The surface areas under the optical density (OD) vs. time growth curves obtained were used to calculate the fractional area f(a), from which the non-inhibitory (NIC) and minimum inhibitory concentrations (MIC) of the antifungal agent were calculated for each fungus using the Gompertz model for decay. Most Aspergillus species showed remarkable resistance to NaMBS, presenting NIC and MIC values higher than 250 and 2500 mg l−1, respectively. The most susceptible fungi were the two Penicillium species and A. carbonarius, which presented very low NIC (<100 mg l−1) and MIC (<1300 mg l−1) values, whereas C. cladosporioides and F. oxysporum presented intermediate values. The method has the advantage of good repeatability and accuracy, rapid results within 48–72 h, growth detection and susceptibility to the antifungal agent for several fungi at the same time, and optimal use of microbiological media by using small volumes of consumables.
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•Fast determination of inhibition concentrations of sodium metabisulphite for different food spoilage fungi•A. carbonarius and Penicillium spp. found to be the more sensitive among fungi tested.•A. flavus and A. tubingensis were the least susceptible species to sodium metabisulphite.
This review elaborates on the significance of Mediterranean raisins, focusing particularly on indigenous Greek varieties (e.g., Zante currants) as a previously overlooked traditional food, currently ...brought on the spotlight, resulting from the increased consumers’ awareness to improve wellness through diet modification. Recent studies on the effect of processing steps on final quality, along with findings on the potential health benefits raisins and currants elicit, are also presented. The development of novel functional food products to further exploit the nutritional value and the bioactive compounds of raisins is evidenced in view of indicating potential food industry applications. Moreover, valorization options of waste and by-product streams obtained from processing facilities are also proposed. Conclusively, raisins and currants should be further enhanced and incorporated in a balanced diet regime through the inclusion in novel foods formulation. Evidently, both the processing of the onset material and side-streams management, are essential to ensure sustainability. Hence, the article also highlights integrated biorefinery approaches, targeting the production of high-value added products that could be re-introduced in the food supply chain and conform with the pillars of bio-economy.
Cheese whey (CW) constitutes a dairy industry by-product, with considerable polluting impact, related mostly with lactose. Numerous bioprocessing approaches have been suggested for lactose ...utilization, however, full exploitation is hindered by strain specificity for lactose consumption, entailing a confined range of end-products. Thus, we developed a CW valorization process generating high added-value products (crude enzymes, nutrient supplements, biopolymers). First, the ability of Aspergillus awamori to secrete β-galactosidase was studied under several conditions during solid-state fermentation (SSF). Maximum enzyme activity (148 U/g) was obtained at 70% initial moisture content after three days. Crude enzymatic extracts were further implemented to hydrolyze CW lactose, assessing the effect of hydrolysis time, temperature and initial enzymatic activity. Complete lactose hydrolysis was obtained after 36 h, using 15 U/mL initial enzymatic activity. Subsequently, submerged fermentations were performed with the produced hydrolysates as onset feedstocks to produce bacterial cellulose (5.6–7 g/L). Our findings indicate a novel approach to valorize CW via the production of crude enzymes and lactose hydrolysis, aiming to unfold the output potential of intermediate product formation and end-product applications. Likewise, this study generated a bio-based material to be further introduced in novel food formulations, elaborating and conforming with the basic pillars of circular economy.
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