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The first complete cloning, purification and characterization of a proteolytic enzyme in Bifidobacterium animalis subsp. lactis to utilize milk proteins and milk-derived peptides are reported. The activity of B. animalis subsp. lactis endopeptidase (PepO) is found with oligopeptide substrates of many amino acid residues and the predominant peptide bond is on the N-terminal side of phenylalanine residues.

Antibiotics have contributed significantly to the improvement of animal health and to the availability of animal-derived foodstuffs. However, the routine use of antimicrobials in animal production has raised concerns not only about the potential presence of residues in foods but, particularly, about the development and spread of antibiotic-resistant bacteria that may compromise the treatment of animal and human infectious diseases. In this context, strategies to reduce antibiotic use and/or mitigate their collateral damage, especially the development of resistance and adverse effects on the host microbiota, must be developed. In this article, different strategies and lines of action or research are proposed to be applied within the framework of the Spanish National Plan for Antibiotic Resistance. [ABSTRACT FROM AUTHOR]

There is an increased interest in secondary plant metabolites, such as polyphenols and carotenoids, due to their proposed health benefits. Much attention has focused on their bioavailability, a prerequisite for further physiological functions. As human studies are time consuming, costly, and restricted by ethical concerns, in vitro models for investigating the effects of digestion on these compounds have been developed and employed to predict their release from the food matrix, bioaccessibility, and assess changes in their profiles prior to absorption. Most typically, models simulate digestion in the oral cavity, the stomach, the small intestine, and, occasionally, the large intestine. A plethora of models have been reported, the choice mostly driven by the type of phytochemical studied, whether the purpose is screening or studying under close physiological conditions, and the availability of the model systems. Unfortunately, the diversity of model conditions has hampered the ability to compare results across different studies. For example, there is substantial variability in the time of digestion, concentrations of salts, enzymes, and bile acids used, pH, the inclusion of various digestion stages; and whether chosen conditions are static (with fixed concentrations of enzymes, bile salts, digesta, and so on) or dynamic (varying concentrations of these constituents). This review presents an overview of models that have been employed to study the digestion of both lipophilic and hydrophilic phytochemicals, comparing digestive conditions in vitro and in vivo and, finally, suggests a set of parameters for static models that resemble physiological conditions. [ABSTRACT FROM AUTHOR]

Food polyphenols are able to selectively modify the growth of susceptible micro-organisms. Wine is a good source of polyphenols and thus, the moderate consumption of this beverage can lead to the modulation of both oral and gut microbiota. This review aims to bring together the knowledge acquired concerning the potential effects of wine polyphenols on human microbiota, as well as taking into account the ability of bacteria to metabolize these compounds. Red wine phenolic composition, characterized by the occurrence of flavan-3-ols, flavonols, anthocyanins, hydroxybenzoic and hydroxycinnamic acids, stilbenes and phenolic alcohols as the main phenolic compounds, will determine the microbiota-modulating effects of wine consumption. Moreover, although the same bacterial genera can be found in oral and gut ecosystems, their relative amount is different, which can influence the metabolic transformations of wine polyphenols. Taking all this into account, the potential implications of these studies on human microbiota are finally discussed together with perspective and future research trends in this field. [Copyright &y& Elsevier]

Abstract: The diversity of the cheese microbiota contributes greatly to the complexity of the cheese-ripening process, crucially important for developing the unique sensory characteristics of each traditional cheese variety. Interactions between microorganisms, e.g. microbial co-operation and antagonistic effects, markedly impact on the survival, growth and activity of microorganisms in cheese. Bacteriocins are antimicrobials that are produced by lactic acid bacteria (LAB) and enable them to dominate the cheese ecosystem. Recently, there has been interest in the possible use of bacteriocins to promote the lysis of LAB and hence contribute to cheese proteolysis and flavour development. This review examines some of the strategies proposed for controlling bacteria in the cheese ecosystem as a means of ensuring high-quality production standards. It focuses on approaches such as the selection of adjuncts, especially facultatively heterofermentative lactobacilli, the selection of new wild strains of LAB and strategies based on microbial co-operation and microbial antagonism. [Copyright &y& Elsevier]

Lactococcin A production in lactococci has recently been linked to a signal-sequence-independent secretory system consisting of a four-gene cluster. Lactococcus lactis subsp. lactis LLM23L-A1 has been obtained after Tn 5 mutagenesis of pLLM23, a plasmid containing the gene cluster responsible for lactococcin A production. In contrast to other Tn 5-generated mutants, strain LLM23L-A1 exhibited a 12-fold increase in lactococcin A production. Overproduction of lactococcin A was not linked to an increased pLLM23 copy number. Restriction-enzyme analysis indicated the site of Tn 5 insertion to be at the 3′ end of lcnD and upstream of the lcnA structural gene. From DNA sequencing, the Tn 5 insertion was located −79 bp upstream of the transcription start site of the lcnA and lciA genes, eliminating eight amino acids from the C-terminal end of lactococcin D. Northern blots revealed overproduction of a 500-base transcript in strain LLM23L-A1, which corresponded to that predicted from the positions of the lactococcin A operon transcriptional start site and the termination structures. This result suggests that the overproduction of lactococcin A in strain LLM23L-A1 is at the transcriptional level and provides further impetus for elucidating the complete regulatory mechanism for lactococcin A expression. [ABSTRACT FROM AUTHOR]

Cites a study examined the effects of various types of animal milk on the characteristics of cheeses, in an effort to optimize the process for making low fat cheeses using ultrafiltration. Information in the ultrafiltration process; Microbiological analyses of the cheeses; Details on the texture of the cheeses; Results of the study.

Caseinomacropeptide (CMP), isolated from cow’s or combined ewe’s and goat’s sweet cheese whey, and whey protein concentrate (WPC), enhanced growth of Bifidobacterium lactis in milk. Supplementation of milk at 2% CMP increased counts of B. lactis by 1.5 log cycles after 24 h incubation at 37 °C when compared with unsupplemented milk, showing no differences between the two CMPs. The highest bacterial growth (9.1 log cfu ml−1) was obtained when milk was supplemented with 2% WPC. The growth-promoting activity shown by WPC could be due to its CMP and whey protein content. CMP and WPC could be utilised to complement milk in order to increase counts of bifidobacteria in probiotic fermented milks. [Copyright &y& Elsevier]

Abstract: Cheese microbiota and their enzymatic conversion of l-methionine to volatile sulphur compounds (VSCs) play an important role in aroma formation during cheese ripening. Here, lactic acid bacteria (LAB) strains isolated from raw goats'' milk cheeses were screened for the major enzymes critical to the formation of VSCs from l-methionine. A large natural biodiversity in enzyme capabilities and high inter- and intra-species variability was found among the LAB isolates investigated. From those isolates tested, lactococci displayed higher C–S lyase specificities towards the sulphur-containing compounds examined than did Lactobacillus and Leuconostoc, in some cases generating higher levels of VSCs than B. linens, known to be an efficient producer of methanethiol (MTL) and related VSCs. Moreover, these differences in C–S lyase activities (determined spectrophotometrically by measuring the formation of free thiol groups) were shown to correspond with the enzymatic potential of the isolates as determined by visualization of enzymatic activities. This technique could therefore prove valuable for the detection and preliminary characterization of C–S lyase activities among LAB isolates. Lactococci were also found to possess higher aminotransferase activities than lactobacilli and leuconostocs, while glutamate dehydrogenase activities were observed to be highest among Leuconostoc and Lactobacillus spp. Meanwhile, α-keto acid decarboxylase activities were highly variable and were measurable in only a limited number of isolates, mainly lactobacilli. From these data, combining indigenous isolates showing high VSCs-producing capabilities with those that facilitate the completion of the metabolic pathway responsible for degrading l-methionine into volatile compounds may provide an efficient approach to enhance cheese aroma development. [Copyright &y& Elsevier]

Abstract: This study describes selective plating methodologies for enumeration of mixed cultures of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, L. acidophilus, L. paracasei subsp. paracasei and Bifidobacterium lactis in fermented milk based on selective antibiotic-free media. Enumeration of S. thermophilus was performed using M17-lactose. MRS-fructose was suitable for enumeration of L. bulgaricus and MRS-maltose for differentiation between L. acidophilus and L. paracasei. The selective enumeration of B. lactis was obtained using MRS-raffinose containing 0.05% LiCl. The bacterial counts obtained using selective methods were equivalent to those under optimum culture conditions at a probability level of 95%. Performance of the methods was verified in fermented milk products where identification of the enumerated species was confirmed by species-specific polymerase chain reaction. This study shows that combination of species-specific polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) analysis has great detection and identification potential for verification of accurate species labelling in fermented milk without prior isolation of the bacteria. [Copyright &y& Elsevier]

Abstract: Several wild lactococcal strains were tested for their ability to produce aroma compounds during growth in milk. Strains were incubated alone and in combination with Lactococcus lactis IFPL730, which is characterized by showing α-keto acid decarboxylase activity. Volatile compounds from incubated milks were analyzed by means of solid-phase microextraction (SPME) and evaluated by gas chromatography–mass spectrometry (GC–MS). Incubated milks were also sniffed for sensory analysis to describe aroma attributes. The combination of L. lactis IFPL326 that showed the highest branched chain aminotransferase activity with IFPL730 contributed to the highest formation of leucine-derived volatile compounds, such as 3-methylbutanal, 3-methyl-1-butanol and 2-hydroxy-4-methyl pentanoic acid methyl ester. In addition, the milk incubated with this combination of strains was awarded, by the test panellists, the highest scores for “ripened cheese” attribute and aroma intensity. The results indicate that combination of L. lactis strains harbouring complementary catabolic routes can contribute to improved cheese aroma formation, the combined cultures with L. lactis IFPL730 resulting in higher volatile compound formation than isolate strains. [Copyright &y& Elsevier]

A set-type fermented milk manufactured from goat''s milk was developed. Optimal curd tension was achieved by supplementation of milk with skim milk powder and whey protein concentrate (WPC). Milk was fermented employing a commercial probiotic starter culture (ABT-2), which contained Streptococcus thermophilus ST-20Y, Lactobacillus acidophilus LA-5, and Bifidobacterium BB-12. Supplementation of milk with 3% WPC reduced fermentation time by 2 h due to the increase in viable counts of S. thermophilus and Bifidobacterium by 0.3 and 0.7 log units, respectively. Addition of WPC increased the protein content (1%) as well as potassium and magnesium content (0.3 and 0.02 g kg−1, respectively). Increase of the protein content led to an increase in the apparent viscosity and gel firmness of the product, and at the same time whey syneresis was reduced. As a consequence, the product received a high score for appearance, taste, aroma, texture and overall acceptance. [Copyright &y& Elsevier]

The safety of the carrageenan (CGN) consumption as a food additive is under debate, with negative effects being associated with the products of hydrolysis of CGN. Moreover, there is an increasing need to integrate gut microbiome analysis in the scientific risk assessment of food additives. The objective of this study was to test the effects of CGN consumption on the gut microbiota and the intestinal homeostasis of young male and female mice. Female and male ICR-CD1 mice (8 weeks old) orally received 540 mg kg -1 day -1 of CGN, representing the maximum-level exposure assessment scenario surveyed for children, over the course of two weeks. Fecal material and peritoneal immune cells were analyzed to determine changes in the fecal microbiota, based on the analysis of bacterial 16S rRNA gene amplicon sequences and short-chain fatty acid (SCFA) concentrations, and some immune functions and redox parameters of peritoneal leukocytes. Non-significant microbiota taxonomical changes associated with CGN intake were found in the mouse stools, resulting the housing time in an increase in bacterial groups belonging to the Bacteroidota phylum. The PICRUSt2 functional predictions showed an overall increase in functional clusters of orthologous genes (COGs) involved in carbohydrate transport and metabolism. A significant increase in the cytotoxicity of fecal supernatants was observed in CGN-fed mice, which correlated with worsening of immune functions and oxidative parameters. The altered immunity and oxidative stress observed in young mice after the consumption of CGN, along with the fecal cytotoxicity shown towards intestinal epithelial cells, may be associated with the gut microbiota's capacity to degrade CGN. The characterization of the gut microbiota's ability to hydrolyze CGN should be included in the risk assessment of this food additive.

Probiotics are valuable microorganisms effective in reducing malnutrition-related infections in children. In this work, a collection of lactobacilli strains representative of traditional Andean fermented beverages was in vitro screened for their capability to survive the gastrointestinal transit, to adhere to the intestinal epithelium and to compete under simulated conditions of the child gut microbiota. The results allowed the selection of the riboflavin overproducing strain Lactiplantibacillus plantarum CECT 9435 based on its good rate of survival under in vitro gastrointestinal conditions when included in a food matrix representing the fortified food supplement Incaparina. The strain also showed good adhesion to HT29 cells producing mucus and outstanding performance in E. coli competition for the adhesion to this epithelial cell line. L. plantarum CECT 9435 gut performance was also evaluated in the child intestinal microbiota simulated in a dynamic gut model (BFBL simulator). The viability of the probiotic candidate in the gut conditions was high during the 7-day intervention period, reaching over 1 × 10 7 counts in each of the reactors simulating the three colonic regions. The transient viability of L. plantarum CECT 9435 within the child gut microbiota and its adhesion capacity to intestinal cells could facilitate the strain potential benefits as probiotic added to fortified supplementary foods destined to malnourished children., (© 2024. The Author(s).)

The gut microbiota should be included in the scientific processes of risk assessment of food additives. Xylitol is a sweetener that shows low digestibility and intestinal absorption, implying that a high proportion of consumed xylitol could reach the colonic microbiota. The present study has evaluated the dose-dependent effects of xylitol intake on the composition and the metabolic activity of the child gut-microbiota. The study was conducted in a dynamic simulator of the colonic microbiota (BFBL Gut Simulator) inoculated with a child pooled faecal sample and supplemented three times per day, for 7 days, with increasing xylitol concentrations (1 g/L, 3 g/L and 5 g/L). Sequencing of 16S rRNA gene amplicons and group-specific quantitative PCR indicated a xylitol dose-response effect on the abundance of Lachnospiraceae, particularly the genera Blautia, Anaerostipes and Roseburia. The microbial changes observed with xylitol corresponded with a dose-dependant effect on the butyrate concentration that, in parallel, favoured an increase in epithelial integrity of Caco-2 cells. The study represents a detailed observation of the bacterial taxa that are the main contributors to the metabolism of xylitol by the child gut microbiota and the results could be relevant in the risk assessment re-evaluation of xylitol as a sweetener., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Opuntia ficus-indica var. Colorada fruit is an important source of indicaxanthin, a betalain with antioxidant, anti-inflammatory, and neuromodulatory potential, proven in both in vitro and in vivo models. Other betalains and phenolic compounds with bioactive activities have also been identified in Colorada fruit extracts. These compounds may degrade by their exposure to different environmental factors, so in the present research, two double emulsion systems (W 1 /O/W 2 ) were elaborated using Tween 20 (TW) and sodium caseinate (SC) as surfactants to encapsulate Colorada fruit pulp extracts, with the aim of enhancing their stability during storage. Encapsulation efficiencies of up to 97.3 ± 2.7%, particle sizes between 236 ± 4 and 3373 ± 64 nm, and zeta potential values of up to ∣46.2∣ ± 0.3 mV were obtained. In addition, the evaluation of the in vitro gastro-intestinal stability and bioaccessibility of the main individual bioactives was carried out by standardized INFOGEST© protocol, obtaining the highest values for the encapsulated extract bioactives in comparison with the non-encapsulated extract (control). Especially, TW double emulsion showed bioaccessibility values of up to 82.8 ± 1.5% for the main bioactives (indicaxanthin, piscidic acid, and isorhamnetin glucoxyl-rhamnosyl-pentoside 2 (IG2)), indicating a promising potential for its use as a functional natural colorant ingredient.

Meniere's disease (MD) is a disorder of the inner ear characterized by chronic episodes of vertigo, tinnitus, increased aural pressure, and sensorineural hearing loss. Causes of MD are unknown, but endolymphatic hydrops is a hallmark. In addition, 5%-15% of MD cases have been identified as familial. Whole-genome sequencing studies of individuals with familial MD identified DTNA and FAM136A as candidate genes for autosomal dominant inheritance of MD. Although the exact roles of these genes in MD are unknown, FAM136A encodes a mitochondrial protein, and DTNA encodes a cytoskeletal protein involved in synapse formation and maintenance, important for maintaining the blood-brain barrier. It is also associated with a particular aquaporin. We tested vestibular and auditory function in dtna and fam136a knockout (KO) mice, using RotaRod and startle reflex-based clicker tests, respectively. Three-factor analysis of variance (ANOVA) results indicated that sex, age, and genotype were significantly correlated with reduced mean latencies to fall ("latencies") for male dtna KO mice, while only age was a significant factor for fam136a KO mice. Fam136a KO mice lost their hearing months before WTs (9-11 months vs. 15-20 months). In male dtna KO mice, divergence in mean latencies compared with other genotypes was first evident at 4 months of age, with older males having an even greater decrease. Our results indicate that fam136a gene mutations generate hearing problems, while dtna gene mutations produce balance deficits. Both mouse models should help to elucidate hearing loss and balance-related symptoms associated with MD., (© 2023 The Authors. European Journal of Neuroscience published by Federation of European Neuroscience Societies and John Wiley & Sons Ltd.)

This study aimed to explore the anti-inflammatory and gut microbiota modulation potentials of flavonoid-rich fraction (PFF) extracted from Passiflora foetida fruits. The results showed that PFF markedly reduced the production of nitric oxide (NO), tumor necrosis factor α (TNF-α), and interleukin 6 (IL-6) in LPS-stimulated RAW 264.7 cells. Meanwhile, PFF treatment also effectively decreased the phosphorylation levels of MAPK, PI3K/Akt, and NF-κB signaling-pathway-related proteins (ERK, JNK, p38, Akt, and p65). Moreover, PFF had an impact on microbial composition and metabolites in a four-stage dynamic simulator of human gut microbiota (BFBL gut model). Specifically, PFF exhibited the growth-promoting ability of several beneficial bacteria, including Bifidobacterium , Enterococcus , Lactobacillus , and Roseburia , and short-chain fatty acid (SCFA) generation ability in gut microbiota. In addition, spectroscopic data revealed that PFF mainly contained five flavonoid compounds, which may be bioactive compounds with anti-inflammatory and gut microbiota modulation potentials. Therefore, PFF could be utilized as a natural anti-inflammatory agent or supplement to health products.

Understanding the individual factors that modulate flavor perception is a central issue for the development of personalized diets strategies to fight obesity. This study aimed to investigate differences in flavor perception between adults with normal weight and those with obesity, as well as some potential biological factors related to these differences. To do that, liking and flavor perception intensity were measured against retronasal olfactory (pineapple, butter, tropical and chocolate) and taste attributes (sweetness, umami and bitter) in 77 individuals grouped as normalweight or obese, according to their body mass index (BMI). Unstimulated saliva was collected from all participants and characterized in terms of salivary flow, total protein content, total antioxidant capacity, total esterase activity and bacterial composition through 16S rDNA amplicon sequencing. The results showed that participants displayed differences in flavor perception according to their BMI group. Thus, the group with obesity showed significant lower liking and intensity scores for low calorie related food aroma (pineapple and tropical), lower taste intensity scores for sweet and umami, and a higher acceptability for umami than the group with normal weight. Significant differences between BMI groups were observed for salivary biochemical variables and specific bacterial taxa, some of which were significantly correlated to flavor intensity. This work suggests for the first time the existence of an oral-brain axis that might contribute to the development or perpetuation of obesity, which opens new and interesting avenues of research., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Meniere's disease (MD) is an inner ear disorder characterised by recurrent vertigo attacks associated with sensorineural hearing loss and tinnitus. Evidence from epidemiology and Whole Exome Sequencing (WES) suggests a genetic susceptibility involving multiple genes, including α-Dystrobrevin ( DTNA ). Here we investigate a Drosophila model. We show that mutation, or knockdown, of the DTNA orthologue in Drosophila , Dystrobrevin ( Dyb ), results in defective proprioception and impaired function of Johnston's Organ (JO), the fly's equivalent of the inner ear. Dyb and another component of the dystrophin-glycoprotein complex (DGC), Dystrophin ( Dys ), are expressed in support cells within JO. Their specific locations suggest that they form part of support cell contacts, thereby helping to maintain the integrity of the hemolymph-neuron diffusion barrier, which is equivalent to a blood-brain barrier. These results have important implications for the human condition, and notably, we note that DTNA is expressed in equivalent cells of the mammalian inner ear., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Requena, Keder, zur Lage, Albert and Jarman.)

Sourdough (SD) fermentation is a traditional biotechnological process used to improve the properties of baked goods. Nowadays, SD fermentation is studied for its potential health effects due to the presence of postbiotic-like components, which refer to a group of inanimate microorganisms and/or their components that confer health benefits on the host. Some postbiotic-like components reported in SD are non-viable microorganisms, short-chain fatty acids, bacteriocins, biosurfactants, secreted proteins/peptides, amino acids, flavonoids, exopolysaccharides, and other molecules. Temperature, pH, fermentation time, and the composition of lactic acid bacteria and yeasts in SD can impact the nutritional and sensory properties of bread and the postbiotic-like effect. Many in vivo studies in humans have associated the consumption of SD bread with higher satiety, lower glycemic responses, increased postprandial concentrations of short-chain fatty acids, and improvement in the symptoms of metabolic or gastrointestinal-related diseases. This review highlights the role of bacteria and yeasts used for SD, the formation of postbiotic-like components affected by SD fermentation and the baking process, and the implications of functional SD bread intake for human health. There are few studies characterizing the stability and properties of postbiotic-like components after the baking process. Therefore, further research is necessary to develop SD bread with postbiotic-related health benefits., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer MT declared a shared parent affiliation with the author TR to the handling editor at the time of review., (Copyright © 2022 Pérez-Alvarado, Zepeda-Hernández, Garcia-Amezquita, Requena, Vinderola and García-Cayuela.)

For developing the recycling of fruit by-products from kiwano, a polysaccharide was extracted from kiwano (Cucumis metuliferus) peels, namely Cucumis metuliferus peels polysaccharide (CMPP), with the aim of investigating the potential beneficial effects. The composition of polysaccharides was analyzed by chemical methods. RAW264.7 macrophages cells and the microbiota dynamics simulator (BFBL gut model) were used for in vitro study. The result showed that CMPP mainly consists of glucuronic acid, arabinose, galactose and rhamnose. By intervening with RAW264.7 cells, CMPP promoted cell proliferation and showed immune-enhancing activity, which significantly (p < 0.05) induced the release of nitric oxide (NO), tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) at a concentration of 50 μg/mL. In addition, CMPP had an impact on the composition of the gut bacteria, increasing the growth of Akkermansia, Bacteroides, Bifidobacterium, Feacalibacterium, and Roseburia. During the intake period, acetic, butyric and propionic acids were all increased, especially (p < 0.05) in the descending colon. Moreover, a decrease in ammonia concentration (10.17 ± 0.50 mM in the ascending colon, 13.21 ± 1.54 mM in the transverse colon and 13.62 ± 0.45 mM in the descending colon, respectively) was observed. In summary, CMPP can be considered as a pectin, showed immunological activity and function of gut microbiota modulation. This study could be the scientific basis of developing kiwano peels as beneficial to human health.