Publications by authors named "Miguel Gueimonde"

147 Publications

Early-Life Development of the Bifidobacterial Community in the Infant Gut.

Int J Mol Sci 2021 Mar 25;22(7). Epub 2021 Mar 25.

Department of Microbiology and Biochemistry of Dairy Products, IPLA-CSIC, 33300 Villaviciosa, Spain.

The establishment of the gut microbiota poses implications for short and long-term health. is an important taxon in early life, being one of the most abundant genera in the infant intestinal microbiota and carrying out key functions for maintaining host-homeostasis. Recent metagenomic studies have shown that different factors, such as gestational age, delivery mode, or feeding habits, affect the gut microbiota establishment at high phylogenetic levels. However, their impact on the specific bifidobacterial populations is not yet well understood. Here we studied the impact of these factors on the different species and subspecies at both the quantitative and qualitative levels. Fecal samples were taken from 85 neonates at 2, 10, 30, 90 days of life, and the relative proportions of the different bifidobacterial populations were assessed by 16S rRNA-23S rRNA internal transcribed spacer (ITS) region sequencing. Absolute levels of the main species were determined by q-PCR. Our results showed that the bifidobacterial population establishment is affected by gestational age, delivery mode, and infant feeding, as it is evidenced by qualitative and quantitative changes. These data underline the need for understanding the impact of perinatal factors on the gut microbiota also at low taxonomic levels, especially in the case of relevant microbial populations such as . The data obtained provide indications for the selection of the species best suited for the development of bifidobacteria-based products for different groups of neonates and will help to develop rational strategies for favoring a healthy early microbiota development when this process is challenged.
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http://dx.doi.org/10.3390/ijms22073382DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036440PMC
March 2021

Longitudinal Study Depicting Differences in Complementary Feeding and Anthropometric Parameters in Late Preterm Infants up to 2 Years of Age.

Nutrients 2021 Mar 18;13(3). Epub 2021 Mar 18.

Department of Functional Biology, University of Oviedo, 33006 Oviedo, Spain.

Ensuring the nutritional demands of preterm (PT) infants during complementary feeding could contribute significantly to the infants' long-term health and development. However, the dietary guidelines for complementary feeding in PT are scarce. Thus, describing dietary intake and identifying nutritional targets for these infants could be of great interest. The aim of this study is to assess the food intake and anthropometric parameters in a Mediterranean infant cohort from 6 to 24 months and to identify nutritional targets especially focused on late preterm infants. This is a longitudinal prospective study analyzing information from administered questionnaires about general characteristics and food frequency consumption in 115 infants (20 PT (32 to 36 gestational weeks), 95 full-term (FT)) at 6, 12 and 24 months of age. Results show that the differences in the prevalence of underweight observed in PT infants vs. FT infants are maintained for up to 6 months of age but disappear at 12 and 24 months. The age of inclusion of new foods and the average intake of the main food groups was not different from that of FTs. Although protein intake at 6 months was directly correlated with weight gain and growth in FT, these associations were not observed in PT. At the nutritional level, the low intake of vitamin D in preterm infants is noteworthy. These findings may be useful when designing new intervention strategies for this population group.
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http://dx.doi.org/10.3390/nu13030982DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8003336PMC
March 2021

Intestinal microbiota alterations by dietary exposure to chemicals from food cooking and processing. Application of data science for risk prediction.

Comput Struct Biotechnol J 2021 29;19:1081-1091. Epub 2021 Jan 29.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA-CSIC), 33300 Villaviciosa, Asturias, Spain.

Diet is one of the main sources of exposure to toxic chemicals with carcinogenic potential, some of which are generated during food processing, depending on the type of food (primarily meat, fish, bread and potatoes), cooking methods and temperature. Although demonstrated in animal models at high doses, an unequivocal link between dietary exposure to these compounds with disease has not been proven in humans. A major difficulty in assessing the actual intake of these toxic compounds is the lack of standardised and harmonised protocols for collecting and analysing dietary information. The intestinal microbiota (IM) has a great influence on health and is altered in some diseases such as colorectal cancer (CRC). Diet influences the composition and activity of the IM, and the net exposure to genotoxicity of potential dietary carcinogens in the gut depends on the interaction among these compounds, IM and diet. This review analyses critically the difficulties and challenges in the study of interactions among these three actors on the onset of CRC. Machine Learning (ML) of data obtained in subclinical and precancerous stages would help to establish risk thresholds for the intake of toxic compounds generated during food processing as related to diet and IM profiles, whereas Semantic Web could improve data accessibility and usability from different studies, as well as helping to elucidate novel interactions among those chemicals, IM and diet.
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http://dx.doi.org/10.1016/j.csbj.2021.01.037DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7892627PMC
January 2021

and environmental enrichment reverse cognitive impairment associated with high-fat high-cholesterol consumption in rats.

Gut Microbes 2021 Jan-Dec;13(1):1-20

Instituto De Neurociencias Del Principado De Asturias (INEUROPA), Asturias, Spain.

Nonalcoholic steatohepatitis (NASH) is one of the most prevalent diseases globally. A high-fat, high-cholesterol (HFHC) diet leads to an early NASH model. It has been suggested that gut microbiota mediates the effects of diet through the microbiota-gut-brain axis, modifying the host's brain metabolism and disrupting cognition. Here, we target NASH-induced cognitive damage by testing the impact of environmental enrichment (EE) and the administration of either GG (LGG) or CIP107961 (AKK). EE and AKK, but not LGG, reverse the HFHC-induced cognitive dysfunction, including impaired spatial working memory and novel object recognition; however, whereas AKK restores brain metabolism, EE results in an overall decrease. Moreover, AKK and LGG did not induce major rearrangements in the intestinal microbiota, with only slight changes in bacterial composition and diversity, whereas EE led to an increase in Firmicutes and Verrucomicrobia members. Our findings illustrate the interplay between gut microbiota, the host's brain energy metabolism, and cognition. In addition, the findings suggest intervention strategies, such as the administration of AKK, for the management of the cognitive dysfunction related to NASH.
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http://dx.doi.org/10.1080/19490976.2021.1880240DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7946069PMC
March 2021

Gut microbes and health.

Gastroenterol Hepatol 2021 Feb 27. Epub 2021 Feb 27.

Ecología Microbiana, Nutrición y Salud, Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Científicas (IATA-CSIC), Valencia, España.

The human body is populated by myriads of microorganisms throughout its surface and in the cavities connected to the outside. The microbial colonisers of the intestine (microbiota) are a functional and non-expendable part of the human organism: they provide genes (microbiome) and additional functions to the resources of our species and participate in multiple physiological processes (somatic development, nutrition, immunity, etc.). Some chronic non-communicable diseases of developed society (atopias, metabolic syndrome, inflammatory diseases, cancer and some behaviour disorders) are associated with dysbiosis: loss of species richness in the intestinal microbiota and deviation from the ancestral microbial environment. Changes in the vertical transmission of the microbiome, the use of antiseptics and antibiotics, and dietary habits in industrialised society appear to be at the origin of dysbiosis. Generating and maintaining diversity in the microbiota is a new clinical target for health promotion and disease prevention.
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http://dx.doi.org/10.1016/j.gastrohep.2021.01.009DOI Listing
February 2021

Resistance of Bifidobacteria Toward Antibiotics.

Methods Mol Biol 2021 ;2278:195-208

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (IPLA-CSIC), Villaviciosa, Spain.

The genus Bifidobacterium constitutes one of the main groups of the human microbiota and some species have a long history of safe consumption supporting an excellent safety record. However, in the context of the increasing worldwide problems associate to the rise of pathogenic microorganisms with acquired resistance to antibiotics, the risk associated to the presence of antibiotic resistance determinants should always be a key starting point for the introduction of any microbial strain into the food chain. Bifidobacteria are not an exception and the presence of resistance to antibiotics is of interest since these microorganisms could potentially act as a reservoir of such resistances. In this context it is necessary to evaluate the presence of antibiotic resistance in any bifidobacterial strain to be included into the food chain. To this end, the first step is the determination of the antibiotic resistance pattern of the strain and the comparison with the susceptibility breakpoints for that species, allowing identifying the presence of atypical resistances in the strain. In this chapter we discuss the many efforts done to harmonize the methods used for the evaluation of antimicrobial susceptibility in the genus Bifidobacterium and the currently available guidelines. Moreover, we describe, in detail, the reference protocols used for evaluating the in vitro antimicrobial activity on bifidobacteria.
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http://dx.doi.org/10.1007/978-1-0716-1274-3_16DOI Listing
April 2021

Selection of Probiotics for Microbiota Modulation in Normal-Weight and Severely Obese Individuals: Focus on Gas Production and Interaction With Intestinal Epithelial Cells.

Front Microbiol 2021 9;12:630572. Epub 2021 Feb 9.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (IPLA-CSIC), Villaviciosa, Spain.

The intestinal microbiota plays important roles in the maintenance of health. Strategies aiming at its modulation, such as probiotics, have received a deal of attention. Several strains have been studied in different models; however, the correlation of results obtained with the data has been limited. This questions the usefulness of such selection models, traditionally relying on over-simplified tests, not considering the influence of the accompanying microbiota or focusing on microbiota composition without considering functional traits. Here we assess the potential of six , and strains in an model to determine their impact on the microbiota not just in terms of composition but also of functionality. Moreover, we compared the responses obtained in two different population groups: normal-weight and severely obese subjects. Fecal cultures were conducted to evaluate the impact of the strains on specific intestinal microbial groups, on the production of short-chain fatty acids, and on two functional responses: the production of gas and the interaction with human intestinal epithelial cells. The response to the different probiotics differed between both human groups. The addition of the probiotic strains did not induce major changes on the microbiota composition, with significant increases detected almost exclusively for the species added. Higher levels of gas production were observed in cultures from normal-weight subjects than in the obese population, with some strains being able to significantly reduce gas production in the latter group. Moreover, in obese subjects all the strains tested and GG were able to modify the response of the intestinal cells, restoring values similar to those obtained with the microbiotas of normal-weight subjects. Our results underline the need for the screening and selection of probiotics in a target-population specific manner by using appropriate models before enrolling in clinical intervention trials.
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http://dx.doi.org/10.3389/fmicb.2021.630572DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7899977PMC
February 2021

Identification of Nutritional Targets in Spanish Children Belonging to the LAyDI Cohort for the Development of Health Promotion Strategies in the First Two Years of Life.

Int J Environ Res Public Health 2021 Jan 22;18(3). Epub 2021 Jan 22.

Area of Physiology, Department of Functional Biology, Faculty of Medicine, University of Oviedo, Julián Claveria, 33006 Oviedo, Spain.

The first 1000 days of life seem to represent the temporal window of opportunity for modulating some of the risk factors associated with the later development of pathologies. Nonetheless, the dietary pattern and nutritional status of children receiving complementary feeding is still understudied. We aimed to assess the food intake in children from the LAyDI cohort (Spain) at 18 and 24 months of age and evaluate this in relation to nutrient requirements and bioactive compound consumption. This was a prospective and multicenter study analyzing information from administered questionnaires about general characteristics and food frequency consumption in 426 children of 18 months and 336 of 24 months. The observed intake of vegetables, fruits, dairy, and eggs was lower than the recommendations in both periods, contrary to the consumption of meat, fish, and pulses. The consumption of energy and macronutrients was similar for all ages studied, with protein intake being slightly higher than the recommended values. Regarding micronutrients, practically the whole sample fell below the vitamin D requirements. In addition, the estimated daily intakes of vitamin E and iron, at 24 months, were below the recommended values for this population group. The mean intake of phenols was around 650 mg/day. Flavanol intake as well as both types of fiber decreased from 18 to 24 months. In conclusion, although these results have to be confirmed in other populations, it seems pertinent to propose the design of nutritional strategies aimed at increasing the intake of vitamins D and E as well as iron in Spanish children up to 2 years.
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http://dx.doi.org/10.3390/ijerph18030939DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7908140PMC
January 2021

Comparison of Different Dietary Indices as Predictors of Inflammation, Oxidative Stress and Intestinal Microbiota in Middle-Aged and Elderly Subjects.

Nutrients 2020 Dec 15;12(12). Epub 2020 Dec 15.

Department of Functional Biology, University of Oviedo, 33006 Oviedo, Spain.

During the last decades the gut microbiota has been identified as a key mediator in the diet-health interaction. However, our understanding on the impact of general diet upon microbiota is still limited. Dietary indices represent an essential approach for addressing the link between diet and health from a holistic point of view. Our aim was to test the predictive potential of seven dietary ratings on biomarkers of inflammation, oxidative stress and on the composition and metabolic activity of the intestinal microbiota. A cross-sectional descriptive study was conducted on a sample of 73 subjects aged >50 years with non-declared pathologies. Dietary inflammatory index (DII), Empirical Dietary Inflammatory Index (EDII), Healthy Eating Index (HEI), Alternative Healthy Eating Index (AHEI), Mediterranean adapted Diet Quality Index-International (DQI-I), Modified Mediterranean Diet Score (MMDS) and relative Mediterranean Diet Score (rMED) were calculated based on a Food Frequency Questionnaire. Major phylogenetic types of the intestinal microbiota were determined by real time polymerase chain reaction (qPCR) and fecal short chain fatty acids (SCFAs) by gas chromatography. While DII, HEI, DQI-I and MMDS were identified as predictors of levels, AHEI and MMDS were negatively associated with group. HEI, AHEI and MMDS were positively associated with fecal SCFAs. In addition, DII and EDII explained lipoperoxidation level and Mediterranean scores the serum IL-8 concentrations. The lower detection of IL-8 in individuals with higher scores on Mediterranean indices may be partially explained by the increased levels of the anti-inflammatory bacterium in such individuals.
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http://dx.doi.org/10.3390/nu12123828DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7765160PMC
December 2020

subsp. CECT7210 ( IM-1) Displays In Vitro Activity against Some Intestinal Pathogens.

Nutrients 2020 Oct 24;12(11). Epub 2020 Oct 24.

Instituto de Productos Lácteos de Asturias (CSIC), P. Río Linares s/n, 33300 Villaviciosa, Spain.

Certain non-digestible oligosaccharides (NDO) are specifically fermented by bifidobacteria along the human gastrointestinal tract, selectively favoring their growth and the production of health-promoting metabolites. In the present study, the ability of the probiotic strain subsp. CECT7210 (herein referred to as IM-1) to utilize a large range of oligosaccharides, or a mixture of oligosaccharides, was investigated. The strain was able to utilize all prebiotics screened. However, galactooligosaccharides (GOS), and GOS-containing mixtures, effectively increased its growth to a higher extent than the other prebiotics. The best synbiotic combination was used to examine the antimicrobial activity against , , and in co-culture experiments. was inhibited by the synbiotic, but it failed to inhibit . Moreover, growth decreased during co-culture with IM-1. Furthermore, adhesion experiments using the intestinal cell line HT29 showed that the strain IM-1 was able to displace some pathogens from the enterocyte layer, especially and , and prevented the adhesion of and . In conclusion, a new synbiotic (probiotic strain IM-1 and GOS) appears to be a potential effective supplement for maintaining infant health. However, further studies are needed to go more deeply into the mechanisms that allow IM-1 to compete with enteropathogens.
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http://dx.doi.org/10.3390/nu12113259DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693895PMC
October 2020

How strong is the evidence that gut microbiota composition can be influenced by lifestyle interventions in a cardio-protective way?

Atherosclerosis 2020 10 11;311:124-142. Epub 2020 Sep 11.

Institute of Biomedicine, Research Centre for Integrative Physiology and Pharmacology, University of Turku, Turku, Finland. Electronic address:

Alterations in composition and function of the gut microbiota have been demonstrated in diseases involving the cardiovascular system, particularly coronary heart disease and atherosclerosis. The data are still limited but the typical altered genera include Roseburia and Faecalibacterium. Plausible mechanisms by which microbiota may mediate cardio-protective effects have been postulated, including the production of metabolites like trimethylamine (TMA), as well as immunomodulatory functions. This raises the question of whether it is possible to modify the gut microbiota by lifestyle interventions and thereby improve cardiovascular health. Nevertheless, lifestyle intervention studies that have involved modifications of dietary intake and/or physical activity, as well as investigating changes in the gut microbiota and subsequent modifications of the cardioprotective markers, are still scarce, and the results have been inconclusive. Current evidence points to benefits of consuming high-fibre foods, nuts and an overall healthy dietary pattern to achieve beneficial effects on both gut microbiota and serum cardiovascular markers, primarily lipids. The relationship between physical exercise and gut microbiota is probably complex and may be dependent on the intensity of exercise. In this article, we review the available evidence on lifestyle, specifically diet, physical activity and smoking as modifiers of the gut microbiota, and subsequently as modifiers of serum cardiovascular health markers. We have attempted to elucidate the plausible mechanisms and further critically appraise the caveats and gaps in the research.
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http://dx.doi.org/10.1016/j.atherosclerosis.2020.08.028DOI Listing
October 2020

Impact of Extreme Obesity and Diet-Induced Weight Loss on the Fecal Metabolome and Gut Microbiota.

Mol Nutr Food Res 2021 03 7;65(5):e2000030. Epub 2020 Oct 7.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Villaviciosa, Asturias, 33300, Spain.

Scope: A limited number of human studies have characterized fecal microbiota and metabolome in extreme obesity and after diet-induced weight loss.

Methods And Results: Fecal samples from normal-weight and extremely obese adults and from obese participants before and after moderate diet-induced weight loss are evaluated for their interaction with the intestinal adenocarcinoma cell line HT29 using an impedance-based in vitro model, which reveals variations in the interaction between the gut microbiota and host linked to obesity status. Microbiota composition, short chain fatty acids, and other intestinal metabolites are further analyzed to assess the interplay among diet, gut microbiota, and host in extreme obesity. Microbiota profiles are distinct between normal-weight and obese participants and are accompanied by fecal signatures in the metabolism of biliary compounds and catecholamines. Moderate diet-induced weight loss promotes shifts in the gut microbiota, and the primary fecal metabolomics features are associated with diet and the gut-liver and gut-brain axes.

Conclusions: Analyses of the fecal microbiota and metabolome enable assessment of the impact of diet on gut microbiota composition and activity, supporting the potential use of certain fecal metabolites or members of the gut microbiota as biomarkers for the efficacy of weight loss in extreme obesity.
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http://dx.doi.org/10.1002/mnfr.202000030DOI Listing
March 2021

The Relationship between Choline Bioavailability from Diet, Intestinal Microbiota Composition, and Its Modulation of Human Diseases.

Nutrients 2020 Aug 5;12(8). Epub 2020 Aug 5.

Instituto de Neurociencias del Principado de Asturias (INEUROPA), 33003 Oviedo, Asturias, Spain.

Choline is a water-soluble nutrient essential for human life. Gut microbial metabolism of choline results in the production of trimethylamine (TMA), which, upon absorption by the host is converted into trimethylamine-N-oxide (TMAO) in the liver. A high accumulation of both components is related to cardiovascular disease, inflammatory bowel disease, non-alcoholic fatty liver disease, and chronic kidney disease. However, the relationship between the microbiota production of these components and its impact on these diseases still remains unknown. In this review, we will address which microbes contribute to TMA production in the human gut, the extent to which host factors (e.g., the genotype) and diet affect TMA production, and the colonization of these microbes and the reversal of dysbiosis as a therapy for these diseases.
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http://dx.doi.org/10.3390/nu12082340DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7468957PMC
August 2020

New players in the relationship between diet and microbiota: the role of macromolecular antioxidant polyphenols.

Eur J Nutr 2021 Apr 27;60(3):1403-1413. Epub 2020 Jul 27.

Department of Functional Biology, University of Oviedo, Oviedo, Spain.

Purpose: Solid evidence has emerged supporting the role of polyphenols and fibers as gut microbiota modulators. These studies have been limited to the data available in food composition databases, which did not include the food content of non-extractable polyphenols (NEPP). The main objective of this work is to quantify the intake of the different types of dietary polyphenols including NEPP and to evaluate their impact on the composition and activity of the intestinal microbiota.

Methods: Cross-sectional descriptive study conducted on a sample of 147 adults with no declared pathologies. Dietary intake has been registered by a semi-quantitative Food Frequency Questionnaire (FFQ) and transformed into extractable (EPP) and NEPP, and dietary fibers based on available databases. Major phylogenetic types of the intestinal microbiota were determined by qPCR and fecal SCFA quantification was performed by gas chromatography.

Results: NEPP account for two-thirds of the total polyphenols intake. A combined analysis by stepwise regression model including all dietary fiber and (poly)phenols has identified hydrolysable (poly)phenol (HPP) intake, as the best predictor of Bacteroides-Prevotella-Porphyromonas group and Bifidobacterium levels in feces. Also, HPPs were positively associated with butyric acid, while insoluble fiber was identified as a predictor of propionic acid in feces.

Conclusion: The intake of macromolecular (poly)phenols could contribute to modulate the gut microbiota by increasing the levels of certain intestinal microorganisms with proven health benefits.
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http://dx.doi.org/10.1007/s00394-020-02339-5DOI Listing
April 2021

An Overview on Fecal Branched Short-Chain Fatty Acids Along Human Life and as Related With Body Mass Index: Associated Dietary and Anthropometric Factors.

Front Microbiol 2020 27;11:973. Epub 2020 May 27.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (IPLA-CSIC), Villaviciosa, Spain.

Short-chain fatty acids (SCFA) are the main bacterial products of the catabolism of carbohydrates and proteins in the gut, and their role is essential in host-microbiota interactions. Acetic, propionic, and butyric acids are the major SCFA produced in the gut, and they have been extensively studied. In contrast, branched short-chain fatty acids (BCFA), mainly isovaleric and isobutyric acids, are produced in less amounts and their fecal levels in different human groups, intestinal microbial producing populations, and influence on health are insufficiently known. They have been proposed as markers of protein fermentation, which leads to the concomitant production of other fermentation products that can be harmful for the colon epithelium. In this context, the aim of this study was to shed light into the production of BCFA by the human intestinal microbiota, as related to age, body mass index (BMI), and diet. Fecal levels of the different SCFA were analyzed by gas chromatography in 232 healthy individuals with ages between 3 months and 95 years, and BMI in adults ranging from 19 to 54. Dietary assessments in adults were obtained through a food frequency questionnaire (FFQ). Molar proportions of BCFA in feces were strongly and positively related with aging. However, not a significant relationship was obtained between BCFA and BMI. A negative correlation was found between the consumption of dietary insoluble fiber and fecal levels of BCFA. More studies are needed for improving our understanding on the relationship of BCFA production profile with the intestinal microbiota composition and human health.
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http://dx.doi.org/10.3389/fmicb.2020.00973DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7271748PMC
May 2020

Donated Human Milk as a Determinant Factor for the Gut Bifidobacterial Ecology in Premature Babies.

Microorganisms 2020 May 19;8(5). Epub 2020 May 19.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA-CSIC), 33300 Villaviciosa, Asturias, Spain.

Correct establishment of the gut microbiome is compromised in premature babies, with being one of the most affected genera. Prematurity often entails the inability to successfully breastfeed, therefore requiring the implementation of other feeding modes; breast milk expression from a donor mother is the recommended option when their own mother's milk is not available. Some studies showed different gut microbial profiles in premature infants fed with breast milk and donor human milk, however, it is not known how this affects the species composition of the genus . The objective of this study was to assess the effect of donated human milk on shaping the gut bifidobacterial populations of premature babies during the first three months of life. We analyzed the gut bifidobacterial communities of 42 premature babies fed with human donor milk or own-mother milk by the 16S rRNA-23S rRNA internal transcriber spaces (ITS) region sequencing and q-PCR. Moreover, metabolic activity was assessed by gas chromatography. We observed a specific bifidobacterial profile based on feeding type, with higher bifidobacterial diversity in the human donor milk group. Differences in specific species composition may contribute to the development of specific new strategies or treatments aimed at mimicking the impact of own-mother milk feeding in neonatal units.
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http://dx.doi.org/10.3390/microorganisms8050760DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7285294PMC
May 2020

Long-Term Coffee Consumption is Associated with Fecal Microbial Composition in Humans.

Nutrients 2020 May 1;12(5). Epub 2020 May 1.

Diet, Microbiota and Health Group, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), 33011 Oviedo, Spain.

Coffee consumption has been related to a preventive effect against several non-transmissible pathologies. Due to the content of this beverage in phytochemicals and minerals, it has been proposed that its impact on health may partly depend on gut microbiota modulation. Our aim was to explore the interaction among gut microbiota, fecal short chain fatty acids, and health-related parameters in 147 healthy subjects classified according to coffee consumption, to deepen the association of the role of the (poly)phenol and alkaloid content of this beverage. Food daily intake was assessed by an annual food frequency questionnaire (FFQ). Coffee consumption was categorized into three groups: non-coffee-consumers (0-3 mL/day), moderate consumers (3-45 mL/day) and high-coffee consumers (45-500 mL/day). Some relevant groups of the gut microbiota were determined by qPCR, and concentration of fecal short chain fatty acids by gas chromatography. Serum health related biomarkers were determined by standardized methods. Interestingly, a higher level of was observed in the high consumers of coffee, who also had lower levels of lipoperoxidation. Two groups of coffee-derived (poly)phenol, methoxyphenols and alkylphenols, and caffeine, among alkaloids, were directly associated with group levels. Thus, regular consumption of coffee appears to be associated with changes in some intestinal microbiota groups in which dietary (poly)phenol and caffeine may play a role.
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http://dx.doi.org/10.3390/nu12051287DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7282261PMC
May 2020

In Vitro Evaluation of Different Prebiotics on the Modulation of Gut Microbiota Composition and Function in Morbid Obese and Normal-Weight Subjects.

Int J Mol Sci 2020 Jan 30;21(3). Epub 2020 Jan 30.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA-CSIC), 33300 Villaviciosa, Asturias, Spain.

The gut microbiota remains relatively stable during adulthood; however, certain intrinsic and environmental factors can lead to microbiota dysbiosis. Its restoration towards a healthy condition using best-suited prebiotics requires previous development of in vitro models for evaluating their functionality. Herein, we carried out fecal cultures with microbiota from healthy normal-weight and morbid obese adults. Cultures were supplemented with different inulin-type fructans (1-kestose, Actilight, P95, Synergy1 and Inulin) and a galactooligosaccharide. Their impact on the gut microbiota was assessed by monitoring gas production and evaluating changes in the microbiota composition (qPCR and 16S rRNA gene profiling) and metabolic activity (gas chromatography). Additionally, the effect on the bifidobacterial species was assessed (ITS-sequencing). Moreover, the functionality of the microbiota before and after prebiotic-modulation was determined in an in vitro model of interaction with an intestinal cell line. In general, 1-kestose was the compound showing the largest effects. The modulation with prebiotics led to significant increases in the group and in obese subjects, whereas in normal-weight individuals, substantial rises in and were appreciated. Notably, the results obtained showed differences in the responses among the tested compounds but also among the studied human populations, indicating the need for developing population-specific products.
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http://dx.doi.org/10.3390/ijms21030906DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7038051PMC
January 2020

Microbiota, Food, and Health.

Int J Mol Sci 2019 Dec 15;20(24). Epub 2019 Dec 15.

Instituto de Productos Lácteos de Asturias (IPLA), CSIC, 33300 Asturias, Spain.

The effects of specific foods, such as products containing probiotics or prebiotics, on human health and the role of intestinal microbiota in this interaction have been a subject of scientific interest for several decades [...].
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http://dx.doi.org/10.3390/ijms20246329DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6940968PMC
December 2019

Use of Fecal Slurry Cultures to Study In Vitro Effects of Bacteriocins on the Gut Bacterial Populations of Infants.

Probiotics Antimicrob Proteins 2020 09;12(3):1218-1225

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (IPLA-CSIC), Villaviciosa, Spain.

Bacteriocins are antimicrobial peptides produced by bacteria to compete with other bacteria for nutrients and ecological niches. The antimicrobial effect of these peptides on the bacterial populations in the gut is likely dynamic as the survival of the microbes in this environment depends on both competition and cooperation. In this study, we evaluated four different bacteriocins from lactic acid bacteria (LAB): nisin, enterocin A (EntA), enterocin K1 (EntK1), and garvicin ML (GarML), which have different inhibition spectra and physicochemical properties. The bacteriocins were tested in vitro using fecal slurry batch cultures from infants. The abundances of some bacterial populations in the cultures were determined using quantitative PCR (qPCR) and the metabolic activity of the gut microbiota was assessed by measuring the production of short-chain fatty acids (SCFA) using gas chromatography. The effects of the bacteriocins correlated well with their antimicrobial spectra and the administered concentrations. Nisin and GarML, with broad antimicrobial spectra, shifted the abundance of several intestinal bacterial groups, while EntA and EntK1, with relative narrower inhibition spectra, showed no or little effect. Moreover, the results from the SCFA analysis were consistent with changes obtained in the bacterial composition. In particular, a reduction in acetate concentration was observed in the samples with low abundance of Bifidobacterium, which is a well-known acetate producer. The variability imposed on the intestinal bacterial populations by the different bacteriocins tested suggests that this type of antimicrobials have great potential to modulate the gut microbiota for medical purposes.
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http://dx.doi.org/10.1007/s12602-019-09614-wDOI Listing
September 2020

Microbiome: Effects of Ageing and Diet.

Curr Issues Mol Biol 2020 27;36:33-62. Epub 2019 Sep 27.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias, Spanish Research Council (IPLA-CSIC), Villaviciosa, Spain.

The microbial community inhabiting our intestine, known as 'microbiota', and the ensemble of their genomes (microbiome) regulate important functions of the host, being essential for health maintenance. The recent development of next-generation sequencing (NGS) methods has greatly facilitated the study of the microbiota and has contributed to evidence of the strong influence exerted by age and diet. However, the precise way in which the diet and its components modify the functionality of the intestinal microbiome is far from being completely known. Changes in the intestinal microbiota occur during ageing, frequently accompanied by physiological changes of the digestive tract, modification of dietary patterns and impairment of the immune system. Establishing nutritional strategies aiming to counterbalance the specific alterations taking place in the microbiota during ageing would contribute to improved health status in the elderly. This review will analyse changes appearing in the intestinal microbiota from adulthood to old age and their association with dietary patterns and lifestyle factors.
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http://dx.doi.org/10.21775/cimb.036.033DOI Listing
September 2020

Neurobehavioral dysfunction in non-alcoholic steatohepatitis is associated with hyperammonemia, gut dysbiosis, and metabolic and functional brain regional deficits.

PLoS One 2019 20;14(9):e0223019. Epub 2019 Sep 20.

Institute of Neurosciences of the Principality of Asturias (INEUROPA), Asturias, Spain.

Non-alcoholic steatohepatitis (NASH) is one of the most prevalent diseases worldwide. While it has been suggested to cause nervous impairment, its neurophysiological basis remains unknown. Therefore, the aim of this study is to unravel the effects of NASH, through the interrelationship of liver, gut microbiota, and nervous system, on the brain and human behavior. To this end, 40 Sprague-Dawley rats were divided into a control group that received normal chow and a NASH group that received a high-fat, high-cholesterol diet. Our results show that 14 weeks of the high-fat, high-cholesterol diet induced clinical conditions such as NASH, including steatosis and increased levels of ammonia. Rats in the NASH group also demonstrated evidence of gut dysbiosis and decreased levels of short-chain fatty acids in the gut. This may explain the deficits in cognitive ability observed in the NASH group, including their depressive-like behavior and short-term memory impairment characterized in part by deficits in social recognition and prefrontal cortex-dependent spatial working memory. We also reported the impact of this NASH-like condition on metabolic and functional processes. Brain tissue demonstrated lower levels of metabolic brain activity in the prefrontal cortex, thalamus, hippocampus, amygdala, and mammillary bodies, accompanied by a decrease in dopamine levels in the prefrontal cortex and cerebellum and a decrease in noradrenalin in the striatum. In this article, we emphasize the important role of ammonia and gut-derived bacterial toxins in liver-gut-brain neurodegeneration and discuss the metabolic and functional brain regional deficits and behavioral impairments in NASH.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0223019PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6754158PMC
March 2020

Fecal Changes Following Introduction of Milk in Infants With Outgrowing Non-IgE Cow's Milk Protein Allergy Are Influenced by Previous Consumption of the Probiotic LGG.

Front Immunol 2019 2;10:1819. Epub 2019 Aug 2.

Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain.

Cow's milk protein allergy (CMPA) is the most common allergy in the first year of life. Non-IgE mediated CMPA is characterized by digestive symptoms and tolerance development before the age of three. Gut microbiota composition in early life has been associated with food allergy. The ingestion of different foods/nutrients may mark different shifts in the microbial colonization of the infant intestine as well as the consumption of probiotics. To analyze changes in microbiota composition and metabolic and cytokine profiles in fecal samples from infants with non-IgE mediated CMPA after successful milk challenges, tolerance acquisition, and increasing dairy introduction in their diet. Twelve children with CMPA, aged between 1 and 2 years old, were recruited for the study. Participants were initially consuming hypoallergenic hydrolyzed formulas (four of them supplemented with the probiotic GG), before being exposed to a standardized oral challenge (SOC) with cow's milk. Fecal samples were collected before, 1 week, and 1 month after performing the SOC. Changes in gut microbiota were determined by high-throughput amplicon sequencing of the 16S rRNA gene. Levels of lactobacilli were also determined by quantitative PCR (qPCR). Microbial metabolites were analyzed by chromatographic methods and fecal cytokines related to the Th1/Th2 balance were determined by immunoassay. Lactic acid bacteria significantly increased in infants who outgrew non-IgE CMPA, after the introduction of milk. Microbial metabolites derived from the fermentation of proteins, such as branched chain fatty acids, and -cresol, diminished. After the SOC, some cytokines related to inflammation (TNF-α, IFN-γ) increased. Accompanying the introduction of an unrestricted diet, we found significant differences in fecal microbial composition, metabolites, and cytokines between infants who did not consume the probiotic GG and those that did. These findings indicate that the introduction of intact milk proteins is followed by modifications in the infant gut environment through changes in immune mediators, microbiota, and its metabolic end-products. Consumption of probiotics during CMPA may contribute to gut homeostasis by fine-tuning these profiles.
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http://dx.doi.org/10.3389/fimmu.2019.01819DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689952PMC
October 2020

Age-Associated Changes in Gut Microbiota and Dietary Components Related with the Immune System in Adulthood and Old Age: A Cross-Sectional Study.

Nutrients 2019 Jul 31;11(8). Epub 2019 Jul 31.

Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Villaviciosa, 33300 Asturias, Spain.

The fecal microbiota plays an important role in human health, and alterations in the microbiota-host interaction seem to be involved in the ageing process. Therefore, it is of interest to develop strategies for promoting a balanced microbiota in old age in order to prevent the physiological and immune decline associated with age. However, the specific microbiota changes in the transition from adulthood to senescence are not yet well understood. Here we assessed the levels of some intestinal microorganisms and short chain fatty acids (SCFAs) across different age-groups. In total, 153 adults from four age groups (<50, 50-65, 66-80, and >80 years-old) were recruited; the levels of different bacterial groups in fecal samples were determined by quantitative polymerase chain reaction (qPCR), and those of SCFA by gas chromatography. Dietary information was collected by using a Food Frequency Questionnaire. The presence of the , , group, and cluster XIVa decreased with age up to 66-80 years of age, with differences reaching statistical significance for the latter group. Interestingly, the levels of some of these microorganisms recovered in the very old age group (>80 years), with these older individuals presenting significantly higher counts of and group than adults and the younger elderly. In addition, ageing was associated with a progressively and statistically significant reduction in the fecal concentrations of SCFAs. Dietary intakes also showed some statistically significant differences among the groups for some macro- and micronutrients. Moreover, associations of some microorganisms with age and macronutrients were also evidenced. Considering the role that fecal microbiota alterations may have in terms of impairing homeostasis and resilience, our results underline the importance of understanding the ageing and immunosenescence processes by including the microbiota perspective.
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http://dx.doi.org/10.3390/nu11081765DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6722604PMC
July 2019

Intestinal Immunomodulation and Shifts on the Gut Microbiota of BALB/c Mice Promoted by Two and Strains Isolated from Human Samples.

Biomed Res Int 2019 18;2019:2323540. Epub 2019 Apr 18.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA-CSIC), 33300 Villaviciosa, Asturias, Spain.

subsp. IPLA 20020 and IPLA 20212, two strains isolated from human samples, were evaluated for safety and influence over the intestinal microbiota and cytokine production by the intestinal tissue of adult BALB/c mice. Mice were divided into four groups receiving during 8 days PBS or a suspension of each strain, prepared fresh or lyophilized (bifidobacteria), at an amount of 4x10 viable cells/day. This dose could be comparable to the probiotic intake of a human adult who consumed about 100-200 mL of functional fermented milk per day, considering the usual level of probiotics in commercial products. No microbial translocation to liver or alterations in food intake, weight, and behavior were observed in treated mice. Intestinal content of secretory immunoglobulin A (s-IgA) was not affected, discarding any adverse effect on the mucosa-associated immunity. The profile of intestinal proinflammatory/regulatory cytokines after intervention evidenced that the microbial strain administered and its cellular state (fresh or lyophilized) as well as the host tissue analyzed (small or large intestine) influenced the immune response and suggests a moderate shift towards a T helper 1 profile (Th1) in the large intestine after the administration of both strains. Changes on relative levels of some intestinal microbial groups were evidenced after intervention. It is noteworthy that butyrate was positively associated with a balanced pro-Th1 immune response. Therefore, subsp IPLA20020 and IPLA 20212 could be considered potential probiotic candidates to be included in functional foods for balancing the intestinal immune response.
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http://dx.doi.org/10.1155/2019/2323540DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6500685PMC
November 2019

Exploring the interactions between serum free fatty acids and fecal microbiota in obesity through a machine learning algorithm.

Food Res Int 2019 07 5;121:533-541. Epub 2018 Dec 5.

Area of Physiology, Department of Functional Biology, Faculty of Medicine, University of Oviedo, C/Julián Clavería s/n, 33006 Oviedo, Asturias, Spain; Group Diet, Microbiota and Health, Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Avda. Roma s/n, 33011 Oviedo, Asturias, Spain. Electronic address:

Serum free fatty acids (FFA) are generally elevated in obesity. The gut microbiota is involved in the host energy metabolism through the regulation of body fat storage, and a link between diet, FFA and the intestinal microbiota seems to exist. Our aim was to explore the interaction among serum FFA levels, gut microbiota, diet and obesity through a model regression tree in 66 subjects (age 52.7 ± 11.2 y) classified according to Body Mass Index (BMI). Total and individual FFA were analyzed by colorimetric enzymatic assay and methyl-tert-butylether-based extraction protocol (MTBE), respectively. Microbiota was determined by qPCR and diet through a food frequency questionnaire. Statistical analyses were performed, and predictive factors for obesity were obtained via classification by decision trees using machine learning methods. An obese-linked FFA profile was characterized by decreased eicosapentaenoic (EPA) and increased linoleic, gamma-linolenic and palmitic acids levels simultaneously. Serum EPA and gender were identified as the most significant variables with 100% and 80% of importance, respectively. Palmitic acid, Bifidobacterium and Faecalibacterium explained >30%, followed by Bacteroides group with 20% and docosahexaenoic acid (DHA) almost with 15% of importance. Also, the regression tree model obtained for predicting obesity, showed a non-obese-linked profile, independently of gender, with serum EPA > 0.235 μg/mL and Bacteroides > 9.055 log n° cells per g of feces. Moreover, Faecalibacterium and Bifidobacterium seemed to play an important role by complementing the levels of FFA in predicting obesity in males and females, respectively.
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http://dx.doi.org/10.1016/j.foodres.2018.12.009DOI Listing
July 2019

Reply: "Letter to the editor Re: Diaz M., et al. 2018, , 1481".

Nutrients 2019 Feb 24;11(2). Epub 2019 Feb 24.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA)-Consejo Superior de Investigaciones Científicas (CSIC), 33300 Villaviciosa-Asturias, Spain.

The objective of this letter of reply is to provide answers to the doubts and critical issues that Martín Martinez and López Liñan [...].
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http://dx.doi.org/10.3390/nu11020476DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6413100PMC
February 2019

The role of yogurt in food-based dietary guidelines.

Nutr Rev 2018 12;76(Suppl 1):29-39

Functional Foods Forum, Faculty of Medicine, University of Turku, Turku, Finland.

In the modern food technology era, one of the aims of food safety and quality is to eliminate or reduce the number of microorganisms in food. This may now be changing. In particular, the importance of live microorganisms as beneficial food constituents is now being recognized. Microorganisms present in food that contribute to the human diet include not only viable bacteria but also metabolites and bioactive components. Yogurt is one of the most biologically active foods consumed by humans. It is an excellent source of proteins, vitamins, and minerals. Additionally, the nutritional value is especially high relative to cost. Potential nutritional benefits are also associated with the ingestion of the bacteria that are ordinarily present at the time of consumption. Thus, yogurt serves as a major source of live bacteria in the human diet, as well as a delivery vehicle for added probiotic bacteria. Yogurt may provide a simple and affordable solution for enhancing the nutritional value of the diet, including the intake of live bacteria and their metabolites. A further benefit may be obtained when yogurt is used as a carrier for specific probiotic bacteria and/or prebiotic compounds. These factors suggest that yogurt could have a more visible role in food-based dietary guidelines.
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http://dx.doi.org/10.1093/nutrit/nuy059DOI Listing
December 2018

Microbiota and Derived Parameters in Fecal Samples of Infants with Non-IgE Cow's Milk Protein Allergy under a Restricted Diet.

Nutrients 2018 Oct 11;10(10). Epub 2018 Oct 11.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA)-Consejo Superior de Investigaciones Científicas (CSIC), 33300 Villaviciosa, Spain.

Cow's milk protein allergy (CMPA) is the most common food allergy in infancy. Non-IgE mediated (NIM) forms are little studied and the responsible mechanisms of tolerance acquisition remain obscure. Our aim was to study the intestinal microbiota and related parameters in the fecal samples of infants with NIM-CMPA, to establish potential links between type of formula substitutes, microbiota, and desensitization. Seventeen infants between one and two years old, diagnosed with NIM-CMPA, were recruited. They were all on an exclusion diet for six months, consuming different therapeutic protein hydrolysates. After this period, stool samples were obtained and tolerance development was evaluated by oral challenges. A control group of 10 age-matched healthy infants on an unrestricted diet were included in the study. Microbiota composition, short-chain fatty acids, calprotectin, and transforming growth factor (TGF)-β₁ levels were determined in fecal samples from both groups. Infants with NIM-CMPA that consumed vegetable protein-based formulas presented microbiota colonization patterns different from those fed with an extensively hydrolyzed formula. Differences in microbiota composition and fecal parameters between NIM-CMPA and healthy infants were observed. Non-allergic infants showed a significantly higher proportion of Bacteroides compared to infants with NIM-CMPA. The type of protein hydrolysate was found to determine gut microbiota colonization and influence food allergy resolution in NIM-CMPA cases.
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http://dx.doi.org/10.3390/nu10101481DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6213916PMC
October 2018

Fecal microbiota profile in a group of myasthenia gravis patients.

Sci Rep 2018 09 26;8(1):14384. Epub 2018 Sep 26.

Department of Microbiology and Biochemistry of Dairy Products, Instituto de Productos Lácteos de Asturias (IPLA-CSIC), 33300, Villaviciosa, Asturias, Spain.

The intestinal microbiota plays a key role in the maintenance of human health. Alterations in this microbiota have been described in several autoimmune diseases, including nervous system diseases. Nevertheless, the information regarding neuromuscular conditions is still limited. In this study, we aimed at characterizing the intestinal microbiota composition in myasthenia gravis patients (MG). To this end fecal samples were taken from ten patients, with antibodies against the acetylcholine receptor, and ten age and sex matched controls from the same population (Asturias region, Spain). Fecal samples were submitted to microbiota analyses by 16S rRNA gene profiling, bifidobacterial ITS-region profiling and qPCR. The fecal levels of short chain fatty acids were determined by gas chromatography. MG patients were found to harbor lower relative proportions of Verrucomicrobiaceae and Bifidobacteriaceae, among others, and increased of the phylum Bacteroidetes and the family Desulfovibrionaceae. The increase of these latter microbial groups was also confirmed at quantitative level by qPCR. In contrast, no statistically significant differences were found between MG patients and the control group in the bifidobacterial population at the species level or in short chain fatty acids profiles. Our data indicates an altered fecal microbiota pattern in MG patients and point out at specific microbiota targets for intervention in this population.
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http://dx.doi.org/10.1038/s41598-018-32700-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6158187PMC
September 2018