Industrial-produced lemon nanovesicles ameliorate experimental colitis-associated damages in rats via the activation of anti-inflammatory and antioxidant responses and microbiota modification.

Plant-derived nanovesicles (PDNVs) have recently emerged as natural delivery systems of biofunctional compounds toward mammalian cells. Considering their already described composition, anti-inflammatory properties, stability, and low toxicity, PDNVs offer a promising path for developing new preventive strategies for several inflammatory diseases, among which the inflammatory bowel disease (IBD). In this study, we explore the protective effects of industrially produced lemon vesicles (iLNVs) in a rat model of IBD. Characterization of iLNVs reveals the presence of small particles less than 200 nm in size and a profile of bioactive compounds enriched in flavonoids and organic acids with known beneficial properties. In vitro studies on human macrophages confirm the safety and anti-inflammatory effects of iLNVs, as evidenced by the reduced expression of pro-inflammatory cytokines and increased levels of anti-inflammatory markers. As evidenced by in vivo experiments, pre-treatment with iLNVs significantly alleviates symptoms and histological features in 2,4 dinitrobenzensulfuric acid (DNBS)-induced colitis in rats. Molecular pathway analysis reveals modulation of NF-κB and Nrf2, indicating anti-inflammatory and antioxidant effects. Finally, iLNVs affects gut microbiota composition, improving the consistent colitis-related alterations. Overall, we demonstrated the protective role of industrially produced lemon nanovesicles against colitis and emphasized their potential in managing IBD through multifaceted mechanisms.

Azithromycin preserves adult hippocampal neurogenesis and behavior in a mouse model of sepsis.

The mammalian hippocampus can generate new neurons throughout life. Known as adult hippocampal neurogenesis (AHN), this process participates in learning, memory, mood regulation, and forgetting. The continuous incorporation of new neurons enhances the plasticity of the hippocampus and contributes to the cognitive reserve in aged individuals. However, the integrity of AHN is targeted by numerous pathological conditions, including neurodegenerative diseases and sustained inflammation. In this regard, the latter causes cognitive decline, mood alterations, and multiple AHN impairments. In fact, the systemic administration of Lipopolysaccharide (LPS) from E. coli to mice (a model of sepsis) triggers depression-like behavior, impairs pattern separation, and decreases the survival, maturation, and synaptic integration of adult-born hippocampal dentate granule cells. Here we tested the capacity of the macrolide antibiotic azithromycin to neutralize the deleterious consequences of LPS administration in female C57BL6J mice. This antibiotic exerted potent neuroprotective effects. It reversed the increased immobility time during the Porsolt test, hippocampal secretion of pro-inflammatory cytokines, and AHN impairments. Moreover, azithromycin promoted the synaptic integration of adult-born neurons and functionally remodeled the gut microbiome. Therefore, our data point to azithromycin as a clinically relevant drug with the putative capacity to ameliorate the negative consequences of chronic inflammation by modulating AHN and hippocampal-related behaviors.

Chronic Consumption of Cocoa Rich in Procyanidins Has a Marginal Impact on Gut Microbiota and on Serum and Fecal Metabolomes in Male Endurance Athletes.

Cocoa is used in the sports world as a supplement, although there is no consensus on its use. We investigated the effect of cocoa intake on intestinal ischemia (intestinal fatty acid-binding protein (I-FABP)), serum lipopolysaccharide (LPS) levels, gastrointestinal symptoms, and gut microbiota in endurance athletes during their training period on an unrestricted diet. We also performed a metabolomics analysis of serum and feces after a bout of exercise before and after supplementation. Cocoa consumption had no effect on I-FABP, LPS, or gastrointestinal symptoms. Cocoa intake significantly increased the abundance of Blautia and Lachnospira genera and decreased the abundance of the Agathobacter genus, which was accompanied by elevated levels of polyphenol fecal metabolites 4-hydroxy-5-(phenyl)-valeric acid and O-methyl-epicatechin-O-glucuronide. Our untargeted approach revealed that cocoa had no significant effects on serum and fecal metabolites and that its consumption had little impact on the metabolome after a bout of physical exercise.

Unraveling Gut Microbiota Signatures Associated with PPARD and PARGC1A Genetic Polymorphisms in a Healthy Population.

Recent studies have revealed the importance of the gut microbiota in the regulation of metabolic phenotypes of highly prevalent metabolic diseases such as obesity, type II diabetes mellitus (T2DM) and cardiovascular disease. Peroxisome proliferator-activated receptors (PPARs) are a family of ligand-activated nuclear receptors that interact with PPAR-γ co-activator-1α (PPARGC1A) to regulate lipid and glucose metabolism. Genetic polymorphisms in PPARD (rs 2267668; A/G) and PPARGC1A (rs 8192678; G/A) are linked to T2DM. We studied the association between the single-nucleotide polymorphisms (SNPs) rs 2267668 and rs 8192678 and microbiota signatures and their relation to predicted metagenome functions, with the aim of determining possible microbial markers in a healthy population. Body composition, physical exercise and diet were characterized as potential confounders. Microbiota analysis of subjects with PPARGC1A (rs 8192678) and PPARD (rs 2267668) SNPs revealed certain taxa associated with the development of insulin resistance and T2DM. Kyoto encyclopedia of gene and genomes analysis of metabolic pathways predicted from metagenomes highlighted an overrepresentation of ABC sugar transporters for the PPARGC1A (rs 8192678) SNP. Our findings suggest an association between sugar metabolism and the PPARGC1A rs 8192678 (G/A) genotype and support the notion of specific microbiota signatures as factors related to the onset of T2DM.

Evaluation of a Zingiber officinale and Bixa orellana Supplement on the Gut Microbiota of Male Athletes: A Randomized Placebo-Controlled Trial.

The gut microbiota has emerged as a factor that influences exercise performance and recovery. The present study aimed to test the effect of a polyherbal supplement containing ginger and annatto called "ReWin(d)" on the gut microbiota of recreational athletes in a pilot, randomized, triple-blind, placebo-controlled trial. Thirty-four participants who practice physical activity at least three times weekly were randomly allocated to two groups, a ReWin(d) group or a maltodextrin (placebo) group. We evaluated the gut microbiota, the production of short-chain fatty acids, and the serum levels of interleukin-6 and lipopolysaccharide at baseline and after 4 weeks. Results showed that ReWin(d) supplementation slightly increased gut microbiota diversity. Pairwise analysis revealed an increase in the relative abundance of Lachnospira (ß-coefficient = 0.013; p = 0.001), Subdoligranulum (ß-coefficient = 0.016; p = 0.016), Roseburia (ß-coefficient = 0.019; p = 0.001), and Butyricicoccus (ß-coefficient = 0.005; p = 0.035) genera in the ReWin(d) group, and a decrease in Lachnoclostridium (ß-coefficient = - 0.008; p = 0.009) and the Christensenellaceae R7 group (ß-coefficient = - 0.010; p < 0.001). Moreover, the Christensenellaceae R-7 group correlated positively with serum interleukin-6 (ρ = 0.4122; p = 0.032), whereas the Lachnospira genus correlated negatively with interleukin-6 (ρ = - 0.399; p = 0.032). ReWin(d) supplementation had no effect on short-chain fatty acid production or on interleukin-6 or lipopolysaccharide levels.

The effect of acute moderate-intensity exercise on the serum and fecal metabolomes and the gut microbiota of cross-country endurance athletes.

Physical exercise can produce changes in the microbiota, conferring health benefits through mechanisms that are not fully understood. We sought to determine the changes driven by exercise on the gut microbiota and on the serum and fecal metabolome using 16S rRNA gene analysis and untargeted metabolomics. A total of 85 serum and 12 fecal metabolites and six bacterial taxa (Romboutsia, Escherichia coli TOP498, Ruminococcaceae UCG-005, Blautia, Ruminiclostridium 9 and Clostridium phoceensis) were modified following a controlled acute exercise session. Among the bacterial taxa, Ruminiclostridium 9 was the most influenced by fecal and serum metabolites, as revealed by linear multivariate regression analysis. Exercise significantly increased the fecal ammonia content. Functional analysis revealed that alanine, aspartate and glutamate metabolism and the arginine and aminoacyl-tRNA biosynthesis pathways were the most relevant modified pathways in serum, whereas the phenylalanine, tyrosine and tryptophan biosynthesis pathway was the most relevant pathway modified in feces. Correlation analysis between fecal and serum metabolites suggested an exchange of metabolites between both compartments. Thus, the performance of a single exercise bout in cross-country non-professional athletes produces significant changes in the microbiota and in the serum and fecal metabolome, which may have health implications.

Key Bacteria in the Gut Microbiota Network for the Transition between Sedentary and Active Lifestyle.

Physical activity modifies the gut microbiota, exerting health benefits on the host; however, the specific bacteria associated with exercise are not yet known. In this work, we propose a novel method, based on hierarchical topology, to study the differences between the microbiota of active and sedentary lifestyles, and to identify relevant bacterial taxa. Our results show that the microbiota network found in active people has a significantly higher overall efficiency and higher transmissibility rate. We also identified key bacteria in active and sedentary networks that could be involved in the conversion of an active microbial network to a sedentary microbial network and vice versa.

Microbiota Features Associated With a High-Fat/Low-Fiber Diet in Healthy Adults.

A high intake of dietary saturated fatty acids (SFAs) is related to an increased risk of obesity, inflammation and cancer-related diseases, and this risk is attenuated only when SFAs are replaced by unsaturated fats and unrefined carbohydrates. The gut microbiota has recently emerged as a new environmental factor in the pathophysiology of these disorders, and is also one of the factors most influenced by diet. We sought to determine whether the gut microbiota of healthy individuals whose intake of SFAs exceeds World Health Organization (WHO) recommendations exhibits features similar to those reported in people with obesity, inflammation, cancer or metabolic disease. Healthy non-obese subjects were divided into two groups based on their SFAs intake. Body composition and gut microbiota composition were analyzed, and associations between bacterial taxa, diet and body fat composition were determined globally and separately by sex. Metagenome functional pathways were predicted by PICRUSt analysis. Subjects whose SFAs intake exceeded WHO recommendations also had a dietary pattern of low fiber intake. This high saturated fat/low fiber diet was associated with a greater sequence abundance of the Anaerotruncus genus, a butyrate producer associated with obesity. Analysis of data of high SFAs intake by sex showed that females presented with a greater abundance of Campylobacter, Blautia, Flavonifractor and Erysipelatoclostridium, whereas males showed higher levels of Anaerotruncus, Eisenbergiella, a genus from the order Clostridiales (FamilyXIIIUCG_001) and two genera from the Lachnospiraceae family. PICRUSt analysis confirmed these data, showing a correlation with a decrease in the abundance of sequences encoding for transporters of some metals such as iron, which is needed to maintain a healthy metabolism. Thus, the microbiota of healthy people on a high SFAs diet contain bacterial taxa (Anaerotruncus, Lachnospiraceae Flavonifractor, Campylobacter, Erysipelotrichacea and Eisenbergiella) that could be related to the development of some diseases, especially obesity and other pro-inflammatory diseases in women. In summary, the present study identifies bacterial taxa that could be considered as early predictors for the onset of different diseases in healthy subjects. Also, sex differences in gut microbiota suggest that women and men differentially benefit from following a specific diet.

Bioinformatic strategies to address limitations of 16rRNA short-read amplicons from different sequencing platforms.

Sequencing the 16S gene rRNA has become a popular method when identifying bacterial communities. However, recent studies address differences in the characterization based on sample preparation, sequencing platforms, and data analysis. In this work, we tested some of the available user-friendly protocols for data analysis with the reads obtained from the sequencing machines Illumina MiSeq and Ion Torrent Personal Genome Machine (PGM). We sought for the advantages and disadvantages of both platforms in terms of accuracy, detected species, and abundance, analyzing a staggered mock community. Four different pipelines were applied: QIIME 1.9.1 with default parameters, QIIME 1.9.1 with modified parameters and chimera removal, VSEARCH 2.3.4, and QIIME 2 v.2018.2. To address the limitations of species level detection we used species-classifier SPINGO. The optimal pipeline for PGM platform, was the use of QIIME 1.9.1 with default parameters (QIIME1), except when a study requires the detection of Bacteroides or other Bacteroidaceae members, in which QIIME1MOD (with chimera removal) seems to be a good alternative. For Illumina Miseq, VSEARCH strategy can be a good option. Our results also confirm that all the tested pipelines can be used for metagenomic analysis at family and genus level.

[Lifestyle modulation of gut microbiota]. / Modulación a través del estilo de vida de la microbiota intestinal.

INTRODUCTION: In recent years, the advance in gut microbiota knowledge has shown that is key in the development and health status of humans. There are many factors that influence the gut microbiota and its balance, being our lifestyle one of the key factors. There is an association between feeding and practicing physical exercise. People who have an active life have a healthier diet, richer in fiber, vegetables and fruits, while sedentary lifestyle is associated with diets with higher fat content and lower fiber. Our feeding behavior and the practice of physical exercise, determine the microbial diversity, as well as the presence of beneficial bacteria for our health. The influence of these factors is determined by the physiological state of the individual (illness/health, obese/lean, young/old), thus more research is needed to determine how changes occur in the microbiota depending on the individual in order to be able to move towards customized nutrition and exercise recommendations according to the needs of each individual.

INTRODUCCIÓN: En los últimos años, el avance en el conocimiento de la microbiota intestinal nos ha demostrado que es clave en el desarrollo y en el estado de salud del ser humano. Son numerosos los factores que influyen sobre la microbiota intestinal y su equilibrio, y nuestro estilo de vida es uno de los factores claves. Existe una asociación entre la alimentación y la práctica de ejercicio físico. Las personas que tienen una vida activa tienen, además, una alimentación más saludable, más rica en fibra, verduras y frutas, mientras que el sedentarismo se asociada al consumo de dietas con mayor contenido de grasa y poca fibra. Nuestro estilo de vida, entendido como alimentación y realización de ejercicio físico, determina la diversidad microbiana, así como la presencia de bacterias beneficiosas para nuestra salud. La influencia de estos factores está determinada por el estado del individuo (enfermedad/salud, obeso/delgado, joven/anciano), por lo que es necesaria más investigación para determinar cómo se producen los cambios en la microbiota en función del individuo, con el fin de poder avanzar hacia una nutrición y unas recomendaciones de ejercicio personalizadas acordes a las necesidades de cada persona.

Can Gut Microbiota and Lifestyle Help Us in the Handling of Anorexia Nervosa Patients?

Gut microbiota is composed of different microorganisms that play an important role in the host. New research shows that bidirectional communications happen between intestinal microbiota and the brain, which is known as the gut⁻brain axis. This communication is significant and could have a negative or positive effect depending on the state of the gut microbiota. Anorexia nervosa (AN) is a mental illness associated with metabolic, immunologic, biochemical, sensory abnormalities, and extremely low body weight. Different studies have shown a dysbiosis in patients with AN. Due to the gut⁻brain axis, it was observed that some of the symptoms could be improved in these patients by boosting their gut microbiota. This paper highlights some evidence connecting the role of microbiota in the AN onset and disease progress. Finally, a proposal is done to include the microbiota analysis as part of the recovery protocol used to treat AN patients. When conducting clinical studies of gut microbiota in AN patients, dysbiosis is expected to be found. Then the prescription of a personalized treatment rich in prebiotics and probiotics could be proposed to reverse the dysbiosis.

A Critical Mutualism - Competition Interplay Underlies the Loss of Microbial Diversity in Sedentary Lifestyle.

Physical exercise improves the overall health status by preventing the development of several diseases. In recent years, it has been observed that physical exercise impacts gut microbiota by increasing the presence of beneficial bacteria and microbial diversity. In contrast, a sedentary lifestyle increases the incidence of chronic diseases that often have an associated loss of microbial diversity. The gut microbiota is a vast ecosystem in which microorganisms interact with each other in different ways; however, microbial ecosystem interactions are scarcely studied. The goal of this study was to determine whether individuals with a sedentary lifestyle have lower diversity in their gut microbiota and how microbial diversity is associated with changes in bacterial network interactions. For that purpose, diet, body composition, physical activity, and sedentarism behavior were characterized for individuals who did or did not comply with the World Health Organization recommendations for physical activity. The composition of the gut microbiome was determined by 16S rRNA gene sequencing. Reorganization of microbial structure with lifestyle was approached from network analysis, where network complexity and the topology of positive and negative interdependences between bacteria were compared and correlated with microbial diversity. Sedentary lifestyle was significantly associated with a diet low in fiber and rich in sugars and processed meat, as well as with high visceral and total corporal fat composition. The diversity (phylogenic diversity, Chao, observed species, and Shannon's index) and network complexity of the gut microbiota were significantly lower in sedentary compared to active individuals. Whereas mutualism or co-occurrence interactions were similar between groups, competitiveness was significantly higher in the active lifestyle group. The mutualism-competition ratio was moderate and positively associated with diversity in sedentary individuals, but not in active individuals. This finding indicates that there is a critical point in this ratio beyond which the stability of the microbial community is lost, inducing a loss of diversity.

Effect of a Protein Supplement on the Gut Microbiota of Endurance Athletes: A Randomized, Controlled, Double-Blind Pilot Study.

Nutritional supplements are popular among athletes to improve performance and physical recovery. Protein supplements fulfill this function by improving performance and increasing muscle mass; however, their effect on other organs or systems is less well known. Diet alterations can induce gut microbiota imbalance, with beneficial or deleterious consequences for the host. To test this, we performed a randomized pilot study in cross-country runners whose diets were complemented with a protein supplement (whey isolate and beef hydrolysate) (n = 12) or maltodextrin (control) (n = 12) for 10 weeks. Microbiota, water content, pH, ammonia, and short-chain fatty acids (SCFAs) were analyzed in fecal samples, whereas malondialdehyde levels (oxidative stress marker) were determined in plasma and urine. Fecal pH, water content, ammonia, and SCFA concentrations did not change, indicating that protein supplementation did not increase the presence of these fermentation-derived metabolites. Similarly, it had no impact on plasma or urine malondialdehyde levels; however, it increased the abundance of the Bacteroidetes phylum and decreased the presence of health-related taxa including Roseburia, Blautia, and Bifidobacterium longum. Thus, long-term protein supplementation may have a negative impact on gut microbiota. Further research is needed to establish the impact of protein supplements on gut microbiota.

Differences in gut microbiota profile between women with active lifestyle and sedentary women.

Physical exercise is a tool to prevent and treat some of the chronic diseases affecting the world's population. A mechanism through which exercise could exert beneficial effects in the body is by provoking alterations to the gut microbiota, an environmental factor that in recent years has been associated with numerous chronic diseases. Here we show that physical exercise performed by women to at least the degree recommended by the World Health Organization can modify the composition of gut microbiota. Using high-throughput sequencing of the 16s rRNA gene, eleven genera were found to be significantly different between active and sedentary women. Quantitative PCR analysis revealed higher abundance of health-promoting bacterial species in active women, including Faecalibacterium prausnitzii, Roseburia hominis and Akkermansia muciniphila. Moreover, body fat percentage, muscular mass and physical activity significantly correlated with several bacterial populations. In summary, we provide the first demonstration of interdependence between some bacterial genera and sedentary behavior parameters, and show that not only does the dose and type of exercise influence the composition of gut microbiota, but also the breaking of sedentary behavior.

Microbial transformation of the diterpene 7-epi-foliol by Fusarium fujikuroi.

The incubation of 3alpha,7alpha,18-trihydroxy-ent-kaur-16-ene (7-epi-foliol) with the fungus Fusarium fujikuroi gave 3alpha,7alpha,18-trihydroxy-ent-kaur-16-en-18-al as the sole product. The biotransformation of other 7alpha- or 7beta-hydroxy derivatives had led to the oxidation of C-19, which is a main step in the biosynthesis of gibberellins and kaurenolides. Now, the presence of the 3alpha-hydroxyl impedes that oxidation, which is directed to the adjacent C-18 hydroxymethyl forming the corresponding aldehyde.

Biotransformation of ent-kaur-16-ene and ent-trachylobane 7ß-acetoxy derivatives by the fungus Gibberella fujikuroi (Fusarium fujikuroi).

Candol A (7ß-hydroxy-ent-kaur-16-ene) (6) is efficiently transformed by Gibberella fujikuroi into the gibberellin plant hormones. In this work, the biotransformation of its acetate by this fungus has led to the formation of 7ß-acetoxy-ent-kaur-16-en-19-oic acid (3), whose corresponding alcohol is a short-lived intermediate in the biosynthesis of gibberellins and seco-ring ent-kaurenoids in this fungus. Further biotransformation of this compound led to the hydroxylation of the 3ß-positions to give 7ß-acetoxy-3ß-hydroxy-ent-kaur-16-en-19-oic acid (14), followed by a 2ß- or 18-hydroxylation of this metabolite. The incubation of epicandicandiol 7ß-monoacetate (7ß-acetoxy-18-hydroxy-ent-kaur-16-ene) (10) produces also the 19-hydroxylation to form the 18,19 diol (20), which is oxidized to give the corresponding C-18 or C-19 acids. These results indicated that the presence of a 7ß-acetoxy group does not inhibit the fungal oxidation of C-19 in 7ß-acetoxy-ent-kaur-16-ene, but avoids the ring B contraction that leads to the gibberellins and the 6ß-hydroxylation necessary for the formation of seco-ring B ent-kaurenoids. The biotransformation of 7ß-acetoxy-ent-trachylobane (trachinol acetate) (27) only led to the formation of 7ß-acetoxy-18-hydroxy-ent-trachylobane (33).

The microbiological transformation of 7alpha-hydroxy-ent-kaur-16-ene derivatives by Gibberella fujikuroi.

The biotransformation of 7alpha-hydroxy-ent-kaur-16-ene (epi-candol A) by the fungus Gibberella fujikuroi gave 7alpha,16alpha,17-trihydroxy-ent-kaur-16-ene and a seco-ring B derivative, fujenoic acid, whilst the incubation of candicandiol (7alpha,18-dihydroxy-ent-kaur-16-ene) and canditriol (7alpha,15alpha,18-trihydroxy-ent-kaur-16-ene) afforded 7alpha,18,19-trihydroxy-ent-kaur-16-ene and 7alpha,11beta,15alpha,18-tetrahydroxy-ent-kaur-16-ene, respectively. The presence of a 7alpha-hydroxyl group in epi-candol A avoids its biotransformation along the biosynthetic pathway of gibberellins, and directs it to the seco-ring B acids route. The 15alpha-hydroxyl group in canditriol inhibits oxidation at C-19 and direct hydroxylation at C-11(beta). The formation of fujenoic acid, from 7alpha-hydroxy-ent-kaur-16-ene, probably occurs via 7alpha-hydroxykaurenoic acid and 7-oxokaurenoic acid, with subsequent hydroxylation at the C-6(beta) position.