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ABSTRACT Gluten possesses unique properties that render it only partially digestible. Consequently, it exerts detrimental effects on a part of the worldwide population who are afflicted with celiac disease (1%) or related disorders (5%), particularly due to the potential for cross-contamination even when adhering to a gluten-free diet (GFD). Finding solutions to break down gluten during digestion has a high nutritional and social impact. Here, a randomized double-blind placebo-controlled in vivo challenge investigated the gluten-degrading activity of a novel probiotic preparation comprising lactobacilli and their cytoplasmic extracts, Bacillus sp., and bacterial protease. In our clinical trial, we collected feces from 70 healthy volunteers at specific time intervals. Probiotic/placebo administration lasted 32 days, followed by 10 days of wash-out. After preliminary GFD to eliminate residual gluten from feces, increasing amounts of gluten (50 mg–10 g) were administered, each one for 4 consecutive days. Compared to placebo, the feces of volunteers fed with probiotics showed much lower amounts of residual gluten, mainly with increased intakes. Probiotics also regulate the intestinal microbial communities, improving the abundance of genera pivotal to maintaining homeostasis. Quantitative PCR confirmed that all probiotics persisted during the intervention, some also during wash-out. Probiotics promoted a fecal metabolome with potential immunomodulating activity, mainly related to derivatives of branched-chain amino acids and short-chain fatty acids.IMPORTANCEThe untapped potential of gluten-degrading bacteria and their application in addressing the recognized limitations of gluten-related disorder management and the ongoing risk of cross-contamination even when people follow a gluten-free diet (GFD) emphasizes the significance of the work. Because gluten, a common protein found in many cereals, must be strictly avoided to stop autoimmune reactions and related health problems, celiac disease and gluten sensitivity present difficult hurdles. However, because of the hidden presence of gluten in many food products and the constant danger of cross-contamination during food preparation and processing, total avoidance is frequently challenging. Our study presents a novel probiotic preparation suitable for people suffering from gluten-related disorders during GFD and for healthy individuals because it enhances gluten digestion and promotes gut microbiota functionality.

Abstract In vitro simulators of the human gastrointestinal (GI) tract are remarkable technological platforms for studying the impact of food on the gut microbiota, enabling continuous and real-time monitoring of key biomarkers. However, comprehensive real-time monitoring of gaseous biomarkers in these systems is required with a cost-effective approach, which has been challenging to perform experimentally to date. In this work, we demonstrate the integration and in-line use of carbon nanotube (CNT)-based chemiresitive gas sensors coated with a thin polydimethylsiloxane (PDMS) membrane for the continuous monitoring of gases within the Simulator of the Human Microbial Ecosystem (SHIME). The findings demonstrate the ability of the gas sensor to continuously monitor the different phases of gas production in this harsh, anaerobic, highly humid, and acidic environment for a long exposure time (16 h) without saturation. This establishes our sensor platform as an effective tool for real-time monitoring of gaseous biomarkers in in vitro systems like SHIME.

Fava bean (Vicia faba L.) is a protein-rich pulse with high nutritional value, but its functional and sensory characteristics limit its application in foods. Solid-state fermentation (SSF) can modify the composition of plant proteins, modulate its functionality, and enhance the sensory aspects. In this study, fava bean flour (FB) was fermented with Aspergillus oryzae and Rhizopus oligosporus to produce FBA and FBR, respectively, ingredients with distinct nutritional, functional, and aroma characteristics. The protein content increased by 20% in FBA and 8% in FBR, while fat levels rose more significantly in FBR (+40%). The overall content of fermentable oligo-, di-, mono-saccharides, and polyols (FODMAPs) decreased by 47% (FBA) and 57% (FBR), although polyol production by A. oryzae was observed. SSF improved the nutritional profile of FBA and FBR, with a notable increase in the concentration of essential amino acids observed, and a reduction in most antinutrients, with the exception of trypsin inhibitors. SSF resulted in the formation of aggregates, which increased the particle size and reduced protein solubility. Emulsions prepared with the fermented ingredients separated faster, and the foaming capacity of both FBA and FBR was decreased, but an increase in water-holding capacity was observed. SSF resulted in the production of predominantly savoury-associated aroma compounds, with compounds characteristic of metallic and mouldy aromas reduced. These results indicate the potential of SSF to transform FB with enhanced nutritional value and improved sensory and functional properties.

This study explored the impact of starter-assisted fermentation on apple blossoms to enhance their potential as a source of antioxidant and antifungal molecules. Fructobacillus fructosus PL22 and Wickerhamomyces anomalus GY1 were chosen as starters owing to their origin and promising ability to modify plant secondary metabolites. An initial assessment through microbiological and physicochemical analyses showed superior outcomes for starter-assisted fermentation compared to the spontaneous process. Enzymatic hydrolysis of proteins, primarily controlled by starters, orchestrated the generation of new low-molecular-weight peptides. W. anomalus GY1 also induced modifications in the phenolic profile, generating a diverse array of bioactive metabolites. These metabolic changes, particularly the release of potentially bioactive peptides, were associated with significant antioxidant activity and marked antifungal efficacy against three common mold species. Our results shed light on the potential of microbial starters to valorize agricultural wastes and convert them into a valuable resource for industry.

Plant protein ingredients are gaining attention for human nutrition, yet they differ significantly from animal proteins in functionality and nutrition. Fungal solid-state fermentation (SSF) can modulate the composition and functionality, increasing their applicability in foods. Quinoa flour (QF) served as a substrate for Aspergillus oryzae and Rhizopus oligosporus, resulting in two fermented ingredients (QFA and QFR) with different nutritional, functional, and aroma characteristics. A higher increase in protein (+35%) and nitrogen (+24%) was observed in the QFA, while fat was predominantly increased in the QFR (+78%). Fermentable oligo-, di-, monosaccharides and polyols (FODMAPs) decreased in the QFR but increased in the QFA due to polyol production. Metabolomic analysis revealed higher lactic acid concentrations in the QFA, and higher citric, malic, and fumaric acid contents in the QFR. The SSF reduced most antinutrients, while R. oligosporus produced saponins. Olfactometry showed the development of fruity ester compounds and a decrease in metallic and cardboard aromas. Both ingredients showed an enhanced water-holding capacity, with the QFA also demonstrating an increased oil-holding capacity. Complex formation increased the particle size, reduced the solubility, and decreased the foaming properties. Mycelium production darkened the ingredients, with the QFR having a higher differential colour index. This study highlights the potential of SSF to produce ingredients with improved nutritional, sensory, and functional properties.

Excessive sugar consumption in young people, who are the major consumers of sugary drinks, combined with limited physical activity, is an important determinant of obesity. Despite their natural appeal, fruit juices have a similar sugar content to that of sugary drinks and once metabolized, they may induce the same biological response. This study aimed to verify whether fermentation processes can make juice consumption healthier and whether reduced-sugar juices have a specific impact on intestinal function. We designed a tailored fermentation of apple-pear juices with lactic acid bacteria and yeasts, which resulted in a reduction of sugar content (27–66%) and caloric intake, and an increase in mannitol content. The impact of newly developed apple-pear juices on gut microbiome composition and functionality was evaluated in vitro using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME). Promising changes were found in the gut microbiota and its metabolic responses and functionality, targeting pathways related to obesity and weight loss (lipopolysaccharide and secondary metabolite biosynthesis, polycyclic aromatic hydrocarbon degradation, and amino sugar and nucleotide sugar metabolism). Additionally, the fermented apple-pear juices positively modulated the intestinal epithelial features. While the simulation of the study simplifies the complex in vivo conditions, it suggests that low-sugar fermented apple-pear juices can elicit targeted responses in the gut ecosystem, contributing to healthier alternatives to traditional fruit juices.

This study investigated the development of hybrid cheese analogues (HCA) made with fermented brewery side-stream ingredients (spent yeast and malt rootlets) and dairy milk. Different percentages of side-stream flours (3.5%, 5%, and 7.5%) were mixed with pasteurized milk, and the developed HCA were evaluated for their biochemical and textural properties. The addition of a fermentation step improved nutrient availability and led to pH (range 4.79–5.60) and moisture content (range 45.86%–61.29%) similar to traditional animal-based fresh cheeses (control). The inclusion of side-stream flours led to coagulation, even without rennet addition. The higher the concentration of the flour used, the faster the coagulation time, suggesting synergistic effect between the enzymes of the rennet and the enzymes present in the fermented side-stream flours. Nevertheless, textural properties were inferior compared to the control. Selected HCA formulations with added 3.5% flour exhibited increased counts of enterococci and enterobacteria cell densities, ranging from 7.28 ± 0.03 to 7.72 ± 0.09 log CFU/g and 4.90 ± 0.16 to 5.41 ± 0.01 log CFU/g, respectively. Compared to the control sample, HCA formulations exhibited higher concentrations of organic acids, peptides, and free amino acids (FAAs). Lactic acid reached up to 23.78 ± 0.94 g/kg of dry matter (DM), while the peptide area reached up to 22918.50 ± 2370.93 mL⋅AU. Additionally, the total concentration of individual FAAs reached up to 2809.74 ± 104.85 mg/kg of DM, contrasted with the control, which resulted in lower concentrations (847.65 ± 0.02 mg/kg of DM). The overall findings suggested that despite challenges in microbiological quality and textural properties, HCA produced with the inclusion of up to 3.5% brewery side-stream flours could be a sustainable solution to produce nutritious dairy alternatives.

Abstract In the current trend where plant‐based foods are preferred over animal‐based foods, pulses represent an alternative source of protein but also of bioactive peptides (BPs). We investigated the pattern of protein hydrolysis during fermentation of red lentils protein isolate (RLPI) with various lactic acid bacteria and yeast strains. Hanseniaspora uvarum SY1 and Fructilactobacillus sanfranciscensis E10 were the most proteolytic microorganisms. H. uvarum SY1 led to the highest antiradical, angiotensin‐converting enzyme‐inhibitory and antifungal activities, as found in low molecular weight water soluble extracts (LMW‐WSE). The 2039 peptide sequences identified by LMW‐WSE were screened using BIOPEP UWM database, and 36 sequences matched with known BPs. Fermentation of RLPI by lactic acid bacteria and yeasts generated 12 peptides undetected in raw RLPI. Besides, H. uvarum SY1 led to the highest abundance (peak areas) of BPs, in particular with antioxidant and ACE‐inhibitory activities. The amino acid sequences LVR and LVL, identified in the fermented RLPI, represent novel findings, as they were detected for the first time in substrates subjected to microbial fermentation. KVI, another BP highly characteristic of RLPI‐SY1, was previously observed only in dried bonito. 44 novel potential BPs, worthy of further characterization, were correlated with antifungal activity.

Spontaneous fermentation of Tritordeum flour enhances the nutritional potential of this hybrid cereal. However, the effect of consumption of Tritordeum sourdough bread (SDB) on gut health remains to be elucidated. This study investigated the effect of in vitro digestion and faecal fermentation of SDB compared to that of traditional baker's yeast (BYB) Tritordeum bread. After 24-h anaerobic faecal fermentation, both SDB and BYB (1% w/v) induced an increase in the relative abundances of Bifidobacterium, Megasphaera, Mitsuokella, and Phascolarctobacterium genera compared to baseline, while concentrations of acetate and butyrate were significantly higher at 24 h for SDB compared to those for BYB. Integrity of intestinal epithelium, as assessed through in vitro trans-epithelial electrical resistance (TEER) assay, was slightly increased after incubation with SDB fermentation supernatants, but not after incubation with BYB fermentation supernatants. The SDB stimulated in vitro mucosal immune response by inducing early secretion of inflammatory cytokines, IL-6 and TNF-α, followed by downregulation of the inflammatory trigger through induction of anti-inflammatory IL-10 expression. Overall, our findings suggest that Tritordeum sourdough can modulate gut microbiota fermentation activity and positively impact the gut health.

Different types of milk are used in the production of milk kefir, but little information is available on the release of potentially antioxidant exopolysaccharides (EPS). The aim of this study was to investigate whether the microbial dynamics and EPS release are dependent on the milk substrate. In our study, the inoculated microbial consortium was driven differently by each type of milk (cow, ewe, and goat). This was evident in the sugar consumption, organic acid production, free amino release, and EPS production. The amount and the composition of the secreted EPS varied depending on the milk type, with implications for the structure and functional properties of the EPS. The low EPS yield in ewe’s milk was associated with a higher lactic acid production and thus with the use of carbon sources oriented towards energy production. Depending on the milk used as substrate, the EPS showed different monosaccharide and FT-IR profiles, microstructures, and surface morphologies. These differences affected the antiradical properties and reducing power of the EPS. In particular, EPS extracted from cow’s milk had a higher antioxidant activity than other milk types, and the antioxidant activity was concentration dependent.

In recent years, heightened attention has been devoted to unravelling the intricate interplay between genetic and environmental factors shaping the gut microbiota and its significance for human health. This study delves into exploring the plausible connection between Alopecia Areata (AA), an autoimmune disease, and the dynamics of the gut microbiome. Examining a cohort of healthy adults and individuals with AA, both the gut microbiota composition and volatile organic compound (VOC) metabolites from faeces and urine were analysed. While overall microbiota composition showed no significant differences, intra-individual variability revealed distinctions related to age, gender, and pathology status, with AA individuals exhibiting reduced species richness and evenness. Differential abundance analysis identified microbial biomarkers for AA, notably Firmicutes, Lachnospirales, and Blautia, while Coprococcus stood out for healthy individuals. The Data Integration Analysis for Biomarker discovery using Latent Components (DIABLO) method further supported these findings including metabolite biomarkers, such as esters of branched chain fatty acids and branched chain amino acids as predictors for AA, suggesting potential links to oxidative stress. Despite certain limitations, the study highlights the complexity of the gut microbiome and its metabolites in the context of AA, while the biomarkers identified could be useful starting points for upcoming studies.

Abstract Background The goal of this study was to create a multi-strain probiotic gel that would foster a lactobacilli-dominated vaginal microbiota in pregnant women and ensure appropriate eubiosis for the newborn. Nomadic lactobacilli (95 strains), mostly isolated from food sources, were preliminarily screened for functional traits before being characterized for their capability to inhibit the two vaginal pathogens Streptococcus agalactiae and Candida albicans, which may lead to adverse pregnancy-related outcomes. Eight best-performing strains were chosen and furtherly investigated for their ability to produce biofilm. Lastly, the two selected potential probiotic candidates were analyzed in vitro for their ability to reduce the inflammation caused by C. albicans infection on the reconstituted human vaginal epithelium (HVE). Results Lactiplantibacillus plantarum produced both isomers of lactic acid, while Lacticaseibacillus paracasei produced only l-isomer. The production of hydrogen peroxide was strain-dependent, with the highest concentrations found within Lact. paracasei strains. The auto-aggregation capacity and hydrophobicity traits were species-independent. S. agalactiae 88II3 was strongly inhibited both at pH 7.0 and 4.0, whereas the inhibition of C. albicans UNIBZ54 was less frequent. Overall, L. plantarum strains had the highest pathogen inhibition and functional scoring. L. plantarum C5 and POM1, which were selected as potential probiotic candidates also based on their ability to form biofilms, were able to counteract the inflammation process caused by C. albicans infection in the HVE model. Conclusions Our multi-step and cumulative scoring-based approach was proven successful in mining and highlighting the probiotic potential of two nomadic lactobacilli strains (L. plantarum C5 and POM1), being applicable to preserve and improve human vaginal health.

Abstract Background In nature, microbial communities undergo changes in composition that threaten their resiliency. Here, we interrogated sourdough, a natural cereal-fermenting metacommunity, as a dynamic ecosystem in which players are subjected to continuous environmental and spatiotemporal stimuli. Results The inspection of spontaneous sourdough metagenomes and transcriptomes revealed dominant, subdominant and satellite players that are engaged in different functional pathways. The highest microbial richness was associated with the highest number of gene copies per pathway. Based on meta-omics data collected from 8 spontaneous sourdoughs and their identified microbiota, we de novo reconstructed a synthetic microbial community SDG. We also reconstructed SMC-SD43 from scratch using the microbial composition of its spontaneous sourdough equivalent for comparison. The KEGG number of dominant players in the SDG was not affected by depletion of a single player, whereas the subdominant and satellite species fluctuated, revealing unique contributions. Compared to SMC-SD43, SDG exhibited broader transcriptome redundancy. The invariant volatilome profile of SDG after in situ long-term back slopping revealed its stability. In contrast, SMC-SD43 lost many taxon members. Dominant, subdominant and satellite players together ensured gene and transcript redundancy. Conclusions Our study demonstrates how, by starting from spontaneous sourdoughs and reconstructing these communities synthetically, it was possible to unravel the metabolic contributions of individual players. For resilience and good performance, the sourdough metacommunity must include dominant, subdominant and satellite players, which together ensure gene and transcript redundancy. Overall, our study changes the paradigm and introduces theoretical foundations for directing food fermentations. Video Abstract

Summary The alteration of a eubiosis status in honeybees’ gut microbiota is directly linked to the occurrence of diseases, and likely to the honeybees decline. Since fructophilic lactobacilli were suggested as symbionts for honeybees, we mechanistically investigated their behaviour under the exposure to agrochemicals (Roundup, Mediator and Reldan containing glyphosate, imidacloprid and chlorpyrifos‐methyl as active ingredients respectively) and plant secondary metabolites (nicotine and p‐coumaric acid) ingested by honeybees as part of their diet. The effects of exposure to agrochemicals and plant secondary metabolites were assessed both on planktonic cells and sessile communities of three biofilm‐forming strains of Apilactobacillus kunkeei. We identified the high sensitivity of A. kunkeei planktonic cells to Roundup and Reldan, while cells embedded in mature biofilms had increased resistance to the same agrochemicals. However, agrochemicals still exerted a substantial inhibitory/control effect if the exposure was during the preliminary steps of biofilm formation. The level of susceptibility resulted to be strain‐specific. Exopolysaccharides resulted in the main component of extracellular polymeric matrix (ECM) in biofilm, but the exposure to Roundup caused a change in ECM production and composition. Nicotine and p‐coumaric acid had a growth‐promoting effect in sessile communities, although no effect was found on planktonic growth.

Summary We proposed a novel phenomic approach to track the effect of short‐term exposures of Lactiplantibacillus plantarum and Leuconostoc pseudomesenteroides to environmental pressure induced by brewers' spent grain (BSG)‐derived saccharides. Water‐soluble BSG‐based medium (WS‐BSG) was chosen as model system. The environmental pressure exerted by WS‐BSG shifted the phenotypes of bacteria in species‐ and strains‐dependent way. The metabolic drift was growth phase‐dependent and likely underlay the diauxic profile of organic acids production by bacteria in response to the low availability of energy sources. Among pentosans, metabolism of arabinose was preferred by L. plantarum and xylose by Leuc. pseudomesenteroides as confirmed by the overexpression of related genes. Bayesian variance analysis showed that phenotype switching towards galactose metabolism suffered the greatest fluctuation in L. plantarum. All lactic acid bacteria strains utilized more intensively sucrose and its plant‐derived isomers. Sucrose‐6‐phosphate activity in Leuc. pseudomesenteroides likely mediated the increased consumption of raffinose. The increased levels of some phenolic compounds suggested the involvement of 6‐phospho‐β‐glucosidases in β‐glucosides degradation. Expression of genes encoding β‐glucoside/cellobiose‐specific EII complexes and phenotyping highlighted an increased metabolism for cellobiose. Our reconstructed metabolic network will improve the understanding of how lactic acid bacteria may transform BSG into suitable food ingredients.

ABSTRACT Raw cow milk is one of the most complex and unpredictable food matrices shaped by the interaction between biotic and abiotic factors. Changes in dairy farming conditions impact the quality and safety of milk, which largely depend on seasonality. Changes in microbiome composition and relative metabolic pathways are derived from microbial interactions, as well as from seasonality, mammary, and extramammary conditions (e.g., farm management and outdoor environment). Breeding data from >600 Apulian farms were examined, and the associated physicochemical parameters were processed by a reductionist approach to obtain a raw cow milk sample subset. We investigated the microbiological variability in cultivable and 16S rRNA sequencing microbiota as affected by seasonal fluctuations at two time points (winter and summer seasons). We identified families (Xanthomonadaceae, Enterobacteriaceae, and Pseudomonadaceae) whose increased abundance during winter may cause a shift toward a pathobiont microbial niche that leads to lower milk quality. Apulian summer season conditions were advantageous to the presence of specific taxa, i.e., Streptococcaceae (i.e., Lactococcus) and Limosilactobacillus fermentum, which in turn may favor better milk preservation. IMPORTANCE The strength of this study lies in the microbiological characterization of a wide range of farm management data to achieve a more comprehensive framework of Apulian milk. Specific regional pedoclimatic and management conditions impact the taxa present and their abundances within this ecological food niche. The obtained results lay the groundwork for comparison with other worldwide extensive farming areas.

Baked goods manufacturing parameters and fermentation conditions interfere with the nutrients content and affect their gastrointestinal fate. Pinsa Romana is a type of pizza that, recently, has been commercially rediscovered and that needed elucidation from a nutritional and digestibility perspective. In this study, six types of Pinsa Romana (five made with indirect method and one produced with straight dough technology) were characterized for their biochemical and nutritional features. Several variables like indirect (biga) Pinsa Romana production process, fermentation time and use of sourdough were investigated. The Pinsa Romana made with biga including sourdough and fermented for 48 h at 16 °C ((PR_48(SD)) resulted in the lowest predicted glycemic index, in the highest content of total peptides, total and individual free amino acids and gamma-amino butyric acid (GABA), and in the best protein quality indexes (protein efficiency ratio and nutritional index). The static in vitro digestion showed that the digesta from PR_48(SD) confirmed a reduced in vitro glycemic response after intake, and it showed a lower bioavailability of hydrophilic peptides. Furthermore, the inclusion of sourdough in biga enhanced the bioavailability of protein-related end-products including human health promoting compounds such as essential amino acids.

ABSTRACT This study provided a new perspective on the bacterial community succession during sauerkraut fermentation, and on resulting metabolic functions. While culture-dependent methods confirmed the key role of the well-known core microbiome species, metagenomic approach (shotgun) revealed Secundilactobacillus malefermentans as a species of the core microbiome, especially during the last weeks of fermentation. Although the potentiality of S. malefermentans has not yet fully explored, it held core functional genes usually attributed to others lactic acid bacteria driving sauerkraut fermentation. Based on our results it is arguable that S. malefermentans might have a key a role during sauerkraut fermentation carried out at low temperature. Under our experimental conditions, the profile of phenolic compounds changed throughout sauerkraut fermentation. The amount of free phenolics, including free phenolic acids, increased at the beginning of the fermentation, whereas the conversion of phenolic acids into microbial derivatives was consistent during the last part of the sauerkraut fermentation. We pioneered correlating changes in the phenolics profile to changes in the microbiome, although the framework presented is still fragmentary. Annotated genes linked to the phenolic compounds metabolism (VprA and padA) were found in many core species during the whole process. A high metabolic potential for phenolics bioconversion emerged for lactobacilli and Pediococcus spp. through correlation analysis between microbiome composition and phenolics profile. IMPORTANCE Our study was not limited to describe the succession pattern of the microbial community during sauerkraut fermentation, but also revealed how some neglected bacterial players belong to the core species during sauerkrauts processing, especially at low temperature. Such species might have a role as potential starters to optimize the fermentation processes and to obtain sauerkrauts with improved and standardized nutritional and sensory features. Furthermore, our correlations between microbiome composition and phenolics profile might also represent new references for sauerkraut biotechnology, aiming to identify new metabolic drivers of potential sauerkraut functionalities. Finally, sauerkraut ecosystem is a tractable model, although with high level of complexity, and resultant ecological information might be extended to other plant ecosystems.

This study aimed to recycle whey milk by-products (protein source) in fruit smoothies (phenolic compounds source) through started-assisted fermentation and delivering sustainable and healthy food formulations capable of providing nutrients that are unavailable due to an unbalanced diet or incorrect eating habits. Five lactic acid bacteria strains were selected as best starters for smoothie production based on the complementarity of pro-technological (kinetics of growth and acidification) traits, exopolysaccharides and phenolics release, and antioxidant activity enhancement. Compared to raw whey milk-based fruit smoothies (Raw_WFS), fermentation led to distinct profiles of sugars (glucose, fructose, mannitol, and sucrose), organic acids (lactic acid and acetic acid), ascorbic acid, phenolic compounds (gallic acid, 3-hydroxybenzoic acid, chlorogenic acid, hydrocaffeic acid, quercetin, epicatechin, procyanidin B2, and ellagic acid) and especially anthocyanins (cyanidin, delphinidin, malvidin, peonidin, petunidin 3-glucoside). Protein and phenolics interaction enhanced the release of anthocyanins, notably under the action of Lactiplantibacillus plantarum. The same bacterial strains outperformed other species in terms of protein digestibility and quality. With variations among starters culture, bio-converted metabolites were most likely responsible for the increase antioxidant scavenging capacity (DPPH, ABTS, and lipid peroxidation) and the modifications in organoleptic properties (aroma and flavor).

Our study investigated the effectiveness of 446 strains of lactic acid bacteria (LAB) belonging to different species and isolated from diverse sources (food, human, and animal) as potential probiotic candidates, with the perspective of producing dietary supplements or pharmacological formulations suitable for enhancing gastrointestinal digestion. The survival capability of all the isolates under harsh gastrointestinal tract conditions was evaluated, in which only 44 strains, named high-resistant, were selected for further food digestibility investigations. All 44 strains hydrolyzed raffinose and exhibited amino and iminopeptidase activities but at various extents, confirming species- and strain-specificity. After partial in vitro digestion mimicking oral and gastric digestive phases, food matrices were incubated with single strains for 24 h. Fermented partially digested matrices provided additional functional properties for some investigated strains by releasing peptides and increasing the release of highly bio-accessible free phenolic compounds. A scoring procedure was proposed as an effective tool to reduce data complexity and quantitively characterize the probiotic potential of each LAB strain, which could be more useful in the selection procedure of powerful probiotics.

Our study proposed date seeds flour (DSF) as an innovative ingredient for sourdough bread production through sustainable bio-recycling. We isolated autochthonous lactic acid bacteria and yeasts from DSF and DSF-derived doughs to build up a reservoir of strains from which to select starters ensuring rapid adaptation and high ecological fitness. The screening based on pro-technological criteria led to the formulation of a mixed starter consisting of Leuconostoc mesenteroides, Lactiplantibacillus plantarum, and Saccharomyces cerevisiae strains, which allowed obtaining a mature type I sourdough after consecutive refreshments, in which an aliquot of the durum wheat flour (DWF) was replaced by DSF. The resulting DSF sourdough and bread underwent an integrated characterization. Sourdough biotechnology was confirmed as a suitable procedure to improve some functional and sensory properties of DWF/DSF mixture formulation. The radical scavenging activity increased due to the consistent release of free phenolics. Perceived bitterness and astringency were considerably diminished, likely because of tannin degradation.

Abstract Background FODMAPs (Fermentable oligosaccharides, disaccharides, monosaccharides, and polyols) intake is associated with the onset of irritable bowel syndrome symptoms. FODMAPs in wheat-derived baked goods may be reduced via bioprocessing by endogenous enzymes and/or microbial fermentation. Because of the inherent enzyme activities, bread made by baker’s yeast and sourdough may result in decreased levels of FODMAPs, whose values are, however, not enough low for people sensitive to FODMAPs. Results Our study investigated the complementary capability of targeted commercial enzymes and metabolically strictly fructophilic lactic acid bacteria (FLAB) to hydrolyze fructans and deplete fructose during wheat dough fermentation. FLAB strains displayed higher fructose consumption rate compared to conventional sourdough lactic acid bacteria. Fructose metabolism by FLAB was faster than glucose. The catabolism of mannitol with the goal of its reuse by FLAB was also investigated. Under sourdough conditions, higher fructans breakdown occurred in FLAB inoculated doughs compared to conventional sourdough bacteria. Preliminary trials allowed selecting Apilactobacillus kunkeei B23I and Fructobacillus fructosus MBIII5 as starter candidates, which were successfully applied in synergy with commercial invertase for low FODMAPs baking. Conclusions Results of this study clearly demonstrated the potential of selected strictly FLAB to strongly reduce FODMAPs in wheat dough, especially under liquid-dough and high oxygenation conditions.

The phenotype-genotype landscape is a projection coming from detailed phenotypic and genotypic data under environmental pressure. Although phenome of microbes or microbial consortia mirrors the functional expression of a genome or set of genomes, metabolic traits rely on the phenotype. Phenomics has the potential to revolution functional genomics. In this review, we discuss why and how phenomics was developed. We described how phenomics may extend our understanding of the assembly of microbial consortia and their functionality, and then we outlined the novel applications within the study of phenomes using Omnilog platform together with a revision of its current application to study lactic acid bacteria (LAB) metabolic traits during food processing. LAB were proposed as a suitable model system to analyze and discuss the implementation and exploitation of this emerging omics approach. We introduced the ‘phenotype switching’, as a new phenotype microarray approach to get insights in bacterial physiology. An overview of methodologies and tools to manage and analyze the generated data was provided. Finally, pro and cons of pipelines developed so far, including the most innovative ones were critically analyzed. We propose an R pipeline, recently deposited, which allows to automatically analyze Omnilog data integrating the latest approaches and implementing the new concepts described here.

This study aimed at investigating the potential of a novel functional herbal tea to act as the carrier for the probiotic Bacillus coagulans GanedenBC30. Spores markedly survived to infusion treatments and the simulation of the gastrointestinal transit only slightly affected the survival, which was 94.8 ± 2.8%. An in-depth investigation using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) which incorporated mucin-covered microcosms was performed. Q-PCR with targeted primers demonstrated that the probiotic quickly colonized mucosal compartments and still persisted after 4 days of wash out. As estimated by plate count combined with 16S rRNA sequencing, B. coagulans GanedenBC30 showed almost the same behavior in luminal compartments both during herbal tea intake and wash out. By summing the luminal and mucosal values and referring to whole volume of colon bioreactors, the content of viable cells of B. coagulans GanedenBC30 was largely above the probiotic threshold.

Autochthonous strains belonging to Lacticaseibacillus paracasei and Lacticaseibacillus casei were screened based on pro-technological (kinetics of growth and acidification, exopolysaccharides biosynthesis), functional (cell viability during processing and storage, in vitro protein digestibility, and in vitro gastrointestinal batch digestion), and sensory properties to ferment milk intended to be included in the ice-cream formulation. The main discrimination among the strains resulted from the sensory evaluation by the panelists, who assigned the highest scores to the ice cream produced with milk fermented with L. casei F14. Antiradical and anti-inflammatory features of such fermented ice cream (with and without the addition of hydroxytyrosol) were shown in Caco-2 cell culture. The pretreatment of Caco-2 cells with fermented ice creams counteracted the accumulation of pro-inflammatory interleukin mediators IL-8 and IL-6 and reactive oxygen species (ROS) induced by detrimental stimuli, and preserved the Caco-2 cell monolayer integrity. The fermentation with L. casei F14 per se played a key role, whereas the inclusion of hydroxytyrosol only contributed to further enhancing the antiradical activity of ice cream. No protective effect on Caco-2 cells was brought about by the addition of dietary fiber.

Although fermentation and hydrolyzation are well-known processes to improve the bioavailability of nutrients and enable the fortification with dietary fibers, the effect of such pre-treatments on the prebiotic features of arabinoxylan-oligosaccharides (AXOS) had not been explored. The middle-term in vitro simulation through the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) demonstrated that the feeding with different formulations (namely oat bran, rye bran and wheat bran) containing hydrolyzed AXOS fermented by lactic acid bacteria significantly increased the synthesis of short-chain fatty acids (SCFA) by colon microbiota, with hydrolyzed and fermented rye bran displaying the highest effect. After two weeks from the interruption of intake, SCFA concentrations significantly decreased but remained still significantly higher compared to the original condition. The microbiome was also affected, with a significant abundance increase in Lactobacillaceae taxon after feeding with all fermented and hydrolyzed formulates. Hydrolyzed and fermented rye bran showed the highest changes. The fungal community, even if it had a lower variety compared to bacteria, was also modulated after feeding with AXOS formulations, with an increase in Candida relative abundance and a decrease in Issatchenkia. On the contrary, the intake of non-hydrolyzed and non-fermented wheat bran did not produce relevant changes of relative abundances. After two weeks from intake interruption (wash out period) such changes were mitigated, and the gut microbiome modulated again to a final structure that was more like the original condition. This finding suggests that hydrolyzed AXOS fermented by lactic acid bacteria could have a more powerful prebiotic effect compared to non-hydrolyzed and non-fermented wheat bran, shaping the colon microbiome and its metabolic answer. However, the intake should be continuous to assure persistent effects. Opening a window into the ecological evolutions and plausible underlying mechanisms, the findings reinforce the perspective to explore more in depth the use of hydrolyzed and fermented AXOS as additional ingredient for bread fortification.

ABSTRACT The contribution of sustainably food processing to healthy intestinal microbial functions is of recent acquisition. The sourdough fermentation fits well with the most sustainable bread making. We manufactured baker’s yeast (BYB) and sourdough (t-SB30) breads, which first underwent to an in-depth characterization. According to nutritional questionnaires, we selected 40 volunteers adhering to the Mediterranean diet. Data on their fecal microbiota and metabolome allowed the selection of two highly representative fecal donors to separately run the Twin Mucosal-SHIME (Twin M-SHIME) under 2-week feeding with BYB and t-SB30. Bread feeding did not affect the microbial composition at phylum and family levels of both donors, in all Twin M-SHIME colon tracts, and lumen and mucosal compartments. The genus core microbiota showed few significant fluctuations, which regarded the relative abundances of Lactobacillus and Leuconostoc according to feeding with BYB and t-SB30, respectively. Compared with BYB, the content of all short chain fatty acids (SCFA), and isovaleric and 2-methylbutyric acids significantly increased with t-SB30 feeding. This was evident for all Twin M-SHIME colon tracts and both donors. The same was found for the content of Asp, Thr, Glu, GABA, and Orn. The bread characterization made possible to identify the main features responsible for this metabolic response. Compared with BYB, t-SB30 had much higher contents of resistant starch, peptides, and free amino acids, and an inhomogeneous microstructure. We used the most efficient approach to investigate a staple food component, excluding interferences from other dietary factors and attenuating human physiology overlaps. The daily consumption of sourdough bread may promote the healthy microbiota metabolism at colon level. IMPORTANCE Knowledge on environmental factors, which may compose the gut microbiota, and drive the host physiology and health is of paramount importance. Human dietary habits and food compositions are pivotal drivers to assemble the human gut microbiota, but, inevitably, unmapped for many diet components, which are poorly investigated individually. Approximately 30% of the human diet consists of fermented foods and beverages. Bread, a fermented/leavened food, is a basic component of the human diet. Its potential effect on gut microbiota composition and functionality is challenging. In this study, we industrially made baker’s yeast and sourdough breads, which were used to feed the Twin Mucosal-SHIME, a worldwide scientifically validated gastrointestinal simulator. Only the consumption of sourdough bread has the potential to enhance the synthesis of short chain fatty acids and free amino acids at the colon level.

Mankai® is a cultivated strain of Wolffia globosa an aquatic plant of the family Lemnaceae commonly known as Duckweeds. Recent studies suggest that consumption of a Mankai® enriched diet may provide positive health effects by decreasing body weight and improving glucose homeostasis and plasma lipid profiles. However, the effects of Mankai® alone on the composition and metabolic output of the human gut microbiota has not been fully investigated. Here, Mankai® was digested and fermented in vitro using a batch culture model of the proximal colon. Inulin and cellulose were used as readily and poorly fermentable control fibers respectively. Mankai® significantly stimulated the production of phenolic metabolites and short chain fatty acids by the gut microbiota (p

Background: Fermented foods are attracting increasing interest due to their nutritional and health benefits, including a positive impact on gut microbiota exerted by their associated microbes. However, information relative to traditional fermented dairy products, along with their autochthonous microbiota, is still fragmented and poorly standardized. Therefore, our aim was to collect and aggregate data useful for obtaining a comprehensive overview translated in a classical database interface that can be easily handled by users. Methods: a preliminary inventory was built up by systematically collecting data from publicly available resources for the creation of a list of traditional dairy foods produced worldwide, including additional metadata useful for stratifying, and collapsing subgroups. Results: we developed the Fermented Dairy Food Database (FDF-DB), a feasible resource comprising 1852 traditional dairy foods (cheeses, fermented milks, and yogurt) for which microbial content and other associated metadata such as geographical indication label, country/region of origin, technological aspects were gathered. Conclusions: FDF-DB is a useful and user-friendly resource where taxonomic information and processing production details converge. This resource will be of great aid for researchers, food industries, stakeholders and any user interested in the identification of technological and microbiological features characterizing traditional fermented dairy products.

Organic farming is gaining a broad recognition as sustainable system, and consumer demand for organic products has increased dramatically in the recent past. Whether organic agriculture delivers overall advantages over conventional agriculture is, however, contentious. Here, the safety, nutritional, and sensory implications of using commercial organic rye, soft, and durum wheat flours rather than conventional-made sourdough bread have been investigated. Culture-dependent and culture-independent approaches were used to explore the microbial architecture of flours and to study their dynamics during sourdough propagation. Besides biochemical features, the main nutritional (amino acid content, asparagine level, and antioxidant activity) characteristics of sourdoughs were investigated, and their effect on the structural, nutritional, and sensory profiles of breads assessed. Overall, the organic farming system led to flours characterized by lower content of asparagine and cell density of Enterobacteriaceae while showing higher concentration of total free amino acids. Differences of the flours mirrored those of sourdoughs and breads. The use of sourdough fermentation guaranteed a further improvement of the flour characteristics; however, a microbial and sensory profile simplification as well as a slight decrease of the biochemical parameters was observed between breads with sourdough after one-cycle fermentation and 10 days of propagation.

A large subset of fermented foods act as vehicles of live environmental microbes, which often contribute food quality assets to the overall diet, such as health-associated microbial metabolites. Foodborne microorganisms also carry the potential to interact with the human gut microbiome via the food chain. However, scientific results describing the microbial flow connecting such different microbiomes as well as their impact on human health, are still fragmented. The aim of this systematic review is to provide a knowledge-base about the scientific literature addressing the connection between foodborne and gut microbiomes, as well as to identify gaps where more research is needed to clarify and map gut microorganisms originating from fermented foods, either traditional or added with probiotics, their possible impact on human gut microbiota composition and to which extent foodborne microbes might be able to colonize the gut environment. An additional aim was also to highlight experimental approaches and study designs which could be better standardized to improve comparative analysis of published datasets. Overall, the results presented in this systematic review suggest that a complex interplay between food and gut microbiota is indeed occurring, although the possible mechanisms for this interaction, as well as how it can impact human health, still remain a puzzling picture. Further research employing standardized and trans-disciplinary approaches aimed at understanding how fermented foods can be tailored to positively influence human gut microbiota and, in turn, host health, are therefore of pivotal importance.

There is an increasing interest in investigating dietary strategies able to modulate the gut microbial ecosystem which, in turn, may play a key role in human health. Dietary fibers (DFs) are widely recognized as molecules with prebiotic effects. The main objective of this systematic review was to: (i) analyze the results available on the impact of DF intervention on short chain fatty acids (SCFAs) production; (ii) evaluate the interplay between the type of DF intervention, the gut microbiota composition and its metabolic activities, and any other health associated outcome evaluated in the host. To this aim, initially, a comprehensive database of literature on human intervention studies assessing the effect of confirmed and candidate prebiotics on the microbial ecosystem was developed. Subsequently, studies performed on DFs and analyzing at least the impact on SCFA levels were extracted from the database. A total of 44 studies from 42 manuscripts were selected for the analysis. Among the different types of fiber, inulin was the DF investigated the most (n = 11). Regarding the results obtained on the ability of fiber to modulate total SCFAs, seven studies reported a significant increase, while no significant changes were reported in five studies, depending on the analytical methodology used. A total of 26 studies did not show significant differences in individual SCFAs, while the others reported significant differences for one or more SCFAs. The effect of DF interventions on the SCFA profile seemed to be strictly dependent on the dose and the type and structure of DFs. Overall, these results underline that, although affecting microbiota composition and derived metabolites, DFs do not produce univocal significant increase in SCFA levels in apparently healthy adults. In this regard, several factors (i.e., related to the study protocols and analytical methods) have been identified that could have affected the results obtained in the studies evaluated. Future studies are needed to better elucidate the relationship between DFs and gut microbiota in terms of SCFA production and impact on health-related markers.

We compared raw bee-collected pollen (Raw-BCP), spontaneously fermented BCP (Unstarted-BCP), and BCP fermented with selected microbial starters (Started-BCP) to deepen whether fermentation may favorably affect the nutrients bioaccessibility and functional features of BCP. Under in vitro gastrointestinal batches, the highest serum-availability of phenolic compounds was found in Started-BCP, highlighting the positive effect exerted by selected microbial starters. The same effect was not found in spontaneously fermented BCP. In colon adenocarcinoma cell line-2 (Caco-2) cells stressed by a pro-inflammatory stimulus, the treatment with Started-BCP halted the increase of pro-inflammatory mediator’s level. Started-BCP counteracted efficiently the deleterious effects of inflammatory stimuli on the integrity of the Caco-2 cells monolayer and its barrier function. Started-BCP successfully counteracted the H2O2-induced intracellular accumulation of reactive oxygen species (ROS) in Caco-2 cells. A protective role against lipopolysaccharide (LPS)-induced inflammation was exerted by Started-BCP in human keratinocytes. The same protective effects on Caco-2 and keratinocyte cell lines were negligible after treatments with Raw-BCP or Unstarted-BCP.

Involvement of the microbiome in many different scalp conditions has been investigated over the years. Studies on the role of the scalp microbiome in specific diseases, such as those involving hair growth alterations like non-cicatricial [androgenetic alopecia (AGA), alopecia areata (AA)] and cicatricial alopecia lichen planopilaris, are of major importance. In the present work, we highlighted the differences in microbial populations inhabiting the scalp of AA subjects and a healthy sample cohort by using an integrated approach relying on metagenomic targeted 16S sequencing analysis, urine metabolomics, and human marker gene expression. Significant differences in genera abundances (p < 0.05) were found in the hypodermis and especially the dermis layer. Based on 16S sequencing data, we explored the differences in predicted KEGG pathways and identified some significant differences in predicted pathways related to the AA pathologic condition such as flagellar, assembly, bacterial chemotaxis, mineral absorption, ABC transporters, cellular antigens, glycosaminoglycan degradation, lysosome, sphingolipid metabolism, cell division, protein digestion and absorption, and energy metabolism. All predicted pathways were significantly enhanced in AA samples compared to expression in healthy samples, with the exceptions of mineral absorption, and ABC transporters. We also determined the expression of TNF-α, FAS, KCNA3, NOD-2, and SOD-2 genes and explored the relationships between human gene expression levels and microbiome composition by Pearson's correlation analysis; here, significant correlations both positive (SOD vs. Staphylococcus, Candidatus Aquiluna) and negative (FAS and SOD2 vs. Anaerococcus, Neisseria, and Acinetobacter) were highlighted. Finally, we inspected volatile organic metabolite profiles in urinary samples and detected statistically significant differences (menthol, methanethiol, dihydrodehydro-beta-ionone, 2,5-dimethylfuran, 1,2,3,4, tetrahydro-1,5,7-trimethylnapthalene) when comparing AA and healthy subject groups. This multiple comparison approach highlighted potential traits associated with AA and their relationship with the microbiota inhabiting the scalp, opening up novel therapeutic interventions in such kind of hair growth disorders mainly by means of prebiotics, probiotics, and postbiotics.

This study aimed to explore common purslane (Portulaca oleracea L.) as potential raw substrate to develop novel functional preparations through lactic acid fermentation. Different lactic acid bacteria were used as single starter. P. oleracea fermented with Lactobacillus kunkeii B7 and Lactobacillus plantarum POM1 resulted in decreased levels (ca. 30%) of oxalic acid, which is commonly recognized as an anti-nutritional compound. Lactobacillus brevis POM4 enhanced by ca. 400% the concentration of γ-amino butyric acid, a non-protein amino acid with various physiological functions. Fermentation of P. oleracea led to a marked increase of α-linalool, a monoterpene compounds with various pharmacological properties. Bioconversion of oleraceins by L. kunkeii B7 is likely responsible for the increased antioxidant activity. The decrease of polyunsaturated fatty acids potentially underlined the synthesis of various derivatives with potential health or nutritional effects. Hence lactic acid fermentation of P. oleracea is a powerful tool to achieve standardized functional foods.

A comprehensive study into the potential of bioprocessing techniques (sprouting and sourdough fermentation) for improving the technological and nutritional properties of wheat breads produced using barley and lentil grains was undertaken. Dextran biosynthesis in situ during fermentation of native or sprouted barley flour (B or SB) alone or by mixing SB flour with native or sprouted lentil flour (SB-L or SB-SL) by Weissella paramesenteroides SLA5, Weissella confusa SLA4, Leuconostoc pseudomesenteroides DSM 20193 or Weissella confusa DSM 20194 was assessed. The acidification and the viscosity increase during 24 h of fermentation with and without 16% sucrose (on flour weight), to promote the dextran synthesis, were followed. After the selection of the fermentation parameters, the bioprocessing was carried out by using Leuconostoc pseudomesenteroides DSM 20193 (the best LAB dextran producer, up to 2.7% of flour weight) and a mixture of SB-SL (30:70% w/w) grains, enabling also the decrease in the raffinose family oligosaccharides. Then, the SB-SL sourdoughs containing dextran or control were mixed with the wheat flour (30% of the final dough) and leavened with baker’s yeast before baking. The use of dextran-containing sourdough allowed the production of bread with structural improvements, compared to the control sourdough bread. Compared to a baker’s yeast bread, it also markedly reduced the predicted glycemic index, increased the soluble (1.26% of dry matter) and total fibers (3.76% of dry matter) content, giving peculiar and appreciable sensory attributes.

Abstract Food and beverage consumption has a great impact on the environment, although there is a lack of information concerning the whole diet. The environmental impact of 153 Italian adults (51 omnivores, 51 ovo-lacto-vegetarians, 51 vegans) and the inter-individual variability within dietary groups were assessed in a real-life context. Food intake was monitored with a 7-d dietary record to calculate nutritional values and environmental impacts (carbon, water, and ecological footprints). The Italian Mediterranean Index was used to evaluate the nutritional quality of each diet. The omnivorous choice generated worse carbon, water and ecological footprints than other diets. No differences were found for the environmental impacts of ovo-lacto-vegetarians and vegans, which also had diets more adherent to the Mediterranean pattern. A high inter-individual variability was observed through principal component analysis, showing that some vegetarians and vegans have higher environmental impacts than those of some omnivores. Thus, regardless of the environmental benefits of plant-based diets, there is a need for thinking in terms of individual dietary habits. To our knowledge, this is the first time environmental impacts of three dietary regimens are evaluated using individual recorded dietary intakes rather than hypothetical diet or diets averaged over a population.

Date fruit (Phoenix dactylifera L.) is a suitable raw matrix for manufacturing of functional ingredients. The aptitude of selected autochthonous lactic acid bacteria for manufacturing of freeze-dried powder from fermented date fruits puree as dietary supplement with health-promoting features was investigated. Fermentation of date fruits puree with selected Lactobacillus plantarum strains allowed the highest concentration of γ-amino butyric acid, conjugated fatty acids, and insoluble dietary fibers. The highest Trolox equivalent antioxidant activity was also induced. Lactic acid fermentation affected the profile of phenolic acids and flavonoids, enriching date fruits puree in phenolic derivatives with high human bioavailability. Fermented date fruits puree was also capable to modulate positively in vitro the growths of several cultivable gut microbial taxa, and enhanced the synthesis of short chain fatty acids. In conclusion, the lactic acid fermentation of date fruits is a suitable tool to achieve a standardized and marketable functional dietary supplement.

Abstract No national legislation anywhere in the world regulates and protects traditional/typical sourdough breads. Sourdough fermentation is firmly associated with a century-old tradition, and with sensory and nutritional quality of breads. A well-defined cell density of lactic acid bacteria has to be reached at the end of fermentation, and be indirectly detectable in baked breads. A Quantitative PCR (qPCR) method was developed to discriminate between breads made with and without sourdoughs. Universal primers targeting an approximately 178-bp fragment of the 16S rRNA-encoding gene of lactic acid bacteria were designed, covering the known diversity of sourdough lactic acid bacteria and excluding commonly encountered flour bacterial contaminants. A total of 191 breads either made with traditional type I and dried sourdough and baker’s yeast, or by a chemical leavening method were shown to be accurately discriminated by means of qPCR. Discriminating values of gene copy number were only weakly correlated with pH values, and with lactate and acetate concentration, thus questioning the validity of these latter indirect indices. The use of sourdough has to be guaranteed to meet both bakery and consumer expectations, and to fulfil legal requirements; our work presents a reliable authentication method providing a suitable tool to satisfy such requirements.

Apple by-products (ABP) underwent fermentation (48 h at 30°C, Fermented-ABP) with a selected binary culture of Weissella cibaria PEP23F and Saccharomyces cerevisiae AN6Y19. Compared to Raw-ABP and Chemically Acidified-ABP (CA-ABP), fermentation markedly increased the hydration properties of ABP. Fermentation led to the highest increases of total and insoluble dietary fibers (DF). Raw-, CA- and Fermented-ABP, at 5 and 10% (w w–1 of flour), were the ingredients for making fortified wheat breads. Addition of ABP and mainly fermentation enhanced dough water absorption and stability, and markedly increased the content of DF. Fortification mainly with 5% of Fermented-ABP did not interfere with bread rheology and color. As shown by profiling volatile compounds, Fermented-ABP imparted agreeable and specific sensory attributes, also appreciated by sensory analysis, and decreased bread hydrolysis index, and delayed mold contamination and firming. Fermented-ABP were suitable ingredients to fortify wheat bread formula, which agreed with bio-economy and environmental sustainability concepts.

This study aimed at characterising the endophytic bacterial communities living in durum wheat roots, as affected by wheat cultivar and inoculation of the Arbuscular mycorrhizal fungus Funneliformis mosseae IMA1 and the wheat root endophytic bacterium Lactobacillus plantarum B.MD.R.A2. These microorganisms were inoculated, alone or in combination, in durum wheat (cultivars Odisseo and Saragolla). Non-inoculated plants of each cultivar represented the controls. Forty-three days after sowing, roots were deprived of the epiphytic microbiota and subjected to DNA extraction. The DNA was used as template in PCR-DGGE analysis of the 16S rRNA gene (variable region V3–V5) and 16S (region V1–V3) metagenetics. Odisseo and Saragolla root endophytic bacterial biotas differed for number of OTUs and composition. In detail, Pseudomonas was higher in Odisseo than in Saragolla. The inoculation of F. mosseae and L. plantarum increased the abundance of Pseudomonas, some Actinobacteria (e.g., Streptomyces, Microbacterium, two genera including several plant growth promoting (PGP) strains) and Bacteroidetes in both cultivars. However, the endophytic bacterial biota of Saragolla roots inoculated just with lactobacilli did not differ from that of the control. The inoculation of Saragolla with F. mosseae, alone or in combination with lactobacilli, led to higher abundance of Rhodococcus, belonging to Actinobacteria and encompassing PGP strains. First, this work showed that F. mosseae and L. plantarum shape the endophytic bacterial biota of durum wheat roots. Abundance of some OTUs was affected by the microbial inoculation, depending on the cultivar. This result represents a starting point for exploitation of beneficial endophytes of wheat roots.

The goal of this study was to investigate the adaptation of L. monocytogenes Scott A cells to treatments with sublethal doses of antimicrobials (ethanol, citral, carvacrol, E-2-hexenal and thyme essential oil). The survival of L. monocytogenes cells was not affected by the antimicrobials at the concentrations assayed, with the exception of ethanol (1% v/v) and thyme essential oil (100 mg/L), which decreased cell viability from 8.53 ± 0.36 to 7.20 ± 0.22 log CFU/mL (P = 0.04). We subsequently evaluated how L. monocytogenes regulates and shapes its proteome in response to antimicrobial compounds. Compared to the control cells grown under optimal conditions, L. monocytogenes treated for 1 h with the antimicrobial compounds showed increased or decreased (≥ or ≤2-fold, respectively, P < 0.05) levels of protein synthesis for 223 protein spots. As shown multivariate clustering analysis, the proteome profiles differed between treatments. Adaptation and shaping of proteomes mainly concerned cell cycle control, cell division, chromosome, motility and regulatory related proteins, carbohydrate, pyruvate, nucleotide and nitrogen metabolism, cofactors and vitamins and stress response with contrasting responses for different stresses. Ethanol, citral (85 mg/l) or (E)-2-hexenal (150 mg/L) adapted cells increased survival during acid stress imposed under model (BHI) and food-like systems.

Although recognized as important sources of functional compounds, milling by-products are often removed from the cereal kernel prior milling process. Indeed, the high presence of fiber in bran and the co-presence of lipids and lipase in germ are often considered as downsides for breadmaking. In this work, Lactobacillus plantarum T6B10 and Weissella confusa BAN8 were used as selected starters to ferment maize milling by-products mixtures made with heat-treated or raw germ and bran. The effects on the biochemical and nutritional features as well as the stability of the milling by-products were investigated. Lactic acid bacteria metabolisms improved the free amino acids and peptides concentrations and the antioxidant activity and caused a relevant phytic acid degradation. Moreover, fermentation allowed a marked decrease of the lipase activity, stabilizing the matrix by preventing oxidative processes. The use of fermented by-products as ingredients improved the nutritional, textural and sensory properties of wheat bread. Fortified breads (containing 25% of fermented by-products) were characterized by a concentration in dietary fiber and proteins of ca. 11 and 13% of dry matter, respectively. Compared to the use of the unfermented ones, the addition of pre-fermented by-products to bread caused a significant increase in protein digestibility (up to 60%), and a relevant decrease of the starch hydrolysis index (ca. 13%). According to the results, this study demonstrates the potential of fermentation to convert maize bran and germ, commonly considered food wastes, into nutritive improvers, meeting nutritional and sensory requests of modern consumers.

Brewers’ spent grain (BSG), the by-product of brewing, was subjected to a xylanase treatment followed by fermentation with Lactiplantibacillus plantarum PU1. Bioprocessed BSG has been used as ingredient to obtain a fortified semolina pasta which can be labeled as “high fiber” and “source of protein” according to the European Community Regulation No. 1924/2006. Compared to native BSG, the use of bioprocessed BSG led to higher protein digestibility and quality indices (essential amino acid index, biological value, protein efficiency ratio, nutritional index), as well as lower predicted glycemic index. Bioprocessing also improved the technological properties of fortified pasta. Indeed, brightfield and confocal laser scanning microscopy revealed the formation of a more homogeneous protein network, resulting from the degradation of the arabinoxylan structure of BSG, and the release of the components entrapped into the cellular compartments. The extensive cell wall disruption contributed to the release of phenols, and conferred enhanced antioxidant activity to the fortified pasta. The persistence of the activity was demonstrated after in vitro-mimicked digestion, evaluating the protective effects of the digested pasta towards induced oxidative stress in Caco-2 cells cultures. The fortified pasta showed a peculiar sensory profile, markedly improved by the pre-treatment, thus confirming the great potential of bioprocessed BSG as health-promoting food ingredient.

Plant matrices are widely recognized as valuable sources of several health promoting compounds [...]

This work aimed to define the microbial consortia that are able to digest gluten into non-toxic and non-immunogenic peptides in the human gastrointestinal tract. Methods: 131 out of 504 tested Bacillus and lactic acid bacteria, specifically Bacillus (64), lactobacilli (63), Pediococcus (1), and Weissella (3), showed strong gastrointestinal resistance and were selected for their PepN, PepI, PepX, PepO, and PepP activities toward synthetic substrates. Based on multivariate analysis, 24 strains were clearly distinct from the other tested strains based on having the highest enzymatic activities. As estimated by RP-HPLC and nano-ESI–MS/MS, 6 cytoplasmic extracts out of 24 selected strains showed the ability to hydrolyze immunogenic epitopes, specifically 57–68 of α9-gliadin, 62–75 of A-gliadin, 134–153 of γ-gliadin, and 57–89 (33-mer) of α2-gliadin. Live and lysed cells of selected strains were combined into different microbial consortia for hydrolyzing gluten under gastrointestinal conditions. Commercial proteolytic enzymes (Aspergillusoryzae E1, Aspergillusniger E2, Bacillussubtilis Veron HPP, and Veron PS proteases) were also added to each microbial consortium. Consortium activity was evaluated by ELISA tests, RP-HPLC-nano-ESI–MS/MS, and duodenal explants from celiac disease patients. Results: two microbial consortia (Consortium 4: Lactiplantibacillus (Lp.) plantarum DSM33363 and DSM33364, Lacticaseibacillus (Lc.) paracasei DSM33373, Bacillussubtilis DSM33298, and Bacilluspumilus DSM33301; and Consortium 16: Lp. plantarum DSM33363 and DSM33364, Lc. paracasei DSM33373, Limosilactobacillusreuteri DSM33374, Bacillusmegaterium DSM33300, B.pumilus DSM33297 and DSM33355), containing commercial enzymes, were able to hydrolyze gluten to non-toxic and non-immunogenic peptides under gastrointestinal conditions. Conclusions: the results of this study provide evidence that selected microbial consortia could potentially improve the digestion of gluten in gluten-sensitive patients by hydrolyzing the immunogenic peptides during gastrointestinal digestion.

Bioprocessing using lactic acid bacteria (LAB) is a powerful means to exploit plant-derived by-products as a food ingredient. LAB have the capability to metabolize a large variety of carbohydrates, but such metabolism only relies on few metabolic routes, conferring on them a high fermentation potential. One example of these pathways is that involving phospho-β-glucosidase genes, which are present in high redundancy within LAB genomes. This enzymatic activity undertakes an ambivalent role during fermentation of plant-based foods related to the release of a wide range of phenolic compounds, from their β-D-glycosylated precursors and the degradation of β-glucopyranosyl derived carbohydrates. We proposed a novel phenomic approach to characterize the metabolism drift of Lactiplantibacillus plantarum and Leuconostoc pseudomesenteroides caused by a lignocellulosic by-product, such as the brewers’ spent grain (BSG), in contrast to Rich De Man, Rogosa and Sharpe (MRS) broth. We observed an increased metabolic activity for gentiobiose, cellobiose and β-glucoside conjugates of phenolic compounds during BSG fermentation. Gene expression analysis confirmed the importance of cellobiose metabolism while a release of lignin-derived aglycones was found during BSG fermentation. We provided a comprehensive view of the important role exerted by LAB 6-phospho-β-glucosidases as well the major metabolic routes undertaken during plant-based fermentations. Further challenges will consider a controlled characterization of pbg gene expression correlated to the metabolism of β-glucosides with different aglycone moieties.

Bee-collected pollen (BCP) is currently receiving increasing attention as a dietary supplement for humans. In order to increase the accessibility of nutrients for intestinal absorption, several biotechnological solutions have been proposed for BCP processing, with fermentation as one of the most attractive. The present study used an integrated metabolomic approach to investigate how the use of starter cultures may affect the volatilome and the profile of bioaccessible phenolics of fermented BCP. BCP fermented with selected microbial starters (Started-BCP) was compared to spontaneously fermented BCP (Unstarted-BCP) and to unprocessed raw BCP (Raw-BCP). Fermentation significantly increased the amount of volatile compounds (VOC) in both Unstarted- and Started-BCP, as well as modifying the relative proportions among the chemical groups. Volatile free fatty acids were the predominant VOC in Unstarted-BCP. Started-BCP was differentiated by the highest levels of esters and alcohols, although volatile free fatty acids were always prevailing. The profile of the VOC was dependent on the type of fermentation, which was attributable to the selected Apilactobacillus kunkeei and Hanseniaspora uvarum strains used as starters, or to the variety of yeasts and bacteria naturally associated to the BCP. Started-BCP and, to a lesser extent, Unstarted-BCP resulted in increased bioaccessible phenolics, which included microbial derivatives of phenolic acids metabolism.

Fresh pasta is subjected to rapid spoilage, mainly due to the metabolic activity of bacteria, yeasts, and especially molds, which negatively affect the sensorial characteristics and the safety of the product. In this work, chickpea flour was fermented with selected lactic acid bacteria, characterized in terms of the antifungal activity, and used to fortify fresh semolina pasta. Pasta was characterized and subjected to a long period of storage after being artificially inoculated with Penicillium roqueforti. Conventional fresh semolina pasta, produced with or without calcium propionate addition, was used as a reference. The water/salt-soluble extract from chickpea sourdough exhibited antifungal activity towards a large spectrum of molds. Its purification led to the identification of ten potentially active peptides. Besides the high content of dietary fibers (4.37%) and proteins (11.20%), nutritional improvements, such as the decrease of the antinutritional factors concentration and the starch hydrolysis index (25% lower than the control) and the increase of the protein digestibility (36% higher than the control), were achieved in fresh pasta fortified with the chickpea sourdough. Inhibition of the indicator mold growth during a 40-day storage period was more effective than in pasta added to calcium propionate.