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1.
Int J Syst Evol Microbiol ; 73(10)2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37882323

RESUMO

A Gram-stain-negative, non-spore-forming, rod-shaped, obligately anaerobic bacterium, designated strain BP5GT, was isolated from the hindgut of a silver drummer (Kyphosus sydneyanus) fish collected from the Hauraki Gulf, New Zealand. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the isolate belonged to the family Lachnospiraceae in the phylum Bacillota and was most closely related to Anaerotignum propionicum with 94.06 % sequence identity. Isolate BP5GT grew on agar medium containing mannitol and fish gut fluid as carbon sources. Clear colonies of approximately 1 mm diameter of the isolate grew within a week at 20-28 °C (optimum, 28 °C) and pH 7.6-8.5 (optimum, pH 8.5). Strain BP5GT was very sensitive to NaCl and the optimal concentration for growth was 0.045 % (w/v). Acetate and propionate were the major fermentation products. The major cellular fatty acids were C12 : 0, C14 : 0, C15 : 0 and C16 : 0. The genome sequence of the isolate was determined. Its G+C content was 38.41 mol% and the 71.41 % average nucleotide identity of the BP5GT genome to its closest neighbour with a sequenced genome (A. propionicum DSM 1682T) indicated low genomic relatedness. Based on the phenotypic and taxonomic characteristics observed in this study, a novel genus and species named Chakrabartyella piscis gen. nov., sp. nov. is proposed for isolate BP5GT (=ICMP 24687T=JCM 35769T).


Assuntos
Ácidos Graxos , Perciformes , Animais , Ácidos Graxos/química , Filogenia , Composição de Bases , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Peixes
3.
Food Funct ; 13(18): 9355-9371, 2022 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-35972507

RESUMO

It is important to understand variability in consumer chewing behavior for designing food products that deliver desired functionalities for target consumer segments. In this study, we selected 29 participants, representing the large range of chewing variation we had observed in 142 healthy young adults, and investigated the influence of chewing behavior on gastrointestinal digestion and colonic fermentation, using in vitro models and brown rice as a model food. Chewing behavior measured by video observations and chewing outcome differed widely between participants, resulting in large differences in the digestibility of carbohydrates. Inter-individual differences in chewing behavior and chewing outcome also significantly affected in vitro patterns of microbial composition and the production of organic acid metabolites, resulting from colonic fermentation, which is increasingly recognized to be important for human health. These digestion/fermentation outcomes were largely related with the chewing time per mouthful, proportion of bolus particles bigger than 2 mm and amount of saliva added to the bolus during chewing. No significant relationships were found with other chewing trajectory and oral physiological measures. These results suggest that modification of chewing may be an effective strategy to control blood glucose levels and to shape gut microbiota and their metabolites, without altering diets, and that further in vivo studies are warranted to confirm these in vitro findings.


Assuntos
Digestão , Mastigação , Glicemia , Fermentação , Alimentos , Humanos , Mastigação/fisiologia , Adulto Jovem
4.
Artigo em Inglês | MEDLINE | ID: mdl-35536223

RESUMO

A Gram-stain-positive, non-spore-forming, rod-shaped, obligately anaerobic bacterium, designated strain BP52GT, was isolated from the hindgut of a Silver Drummer (Kyphosus sydneyanus) fish collected from the Hauraki Gulf, New Zealand. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that the isolate belonged to the family Erysipelotrichaceae in the phylum Firmicutes and was most closely related to Clostridium saccharogumia with 93.3 % sequence identity. Isolate BP52GT grew on agar medium containing mannitol as the sole carbon source. White, opaque and shiny colonies of the isolate measuring approximately 1 mm diameter grew within a week at 20-28 °C (optimum, 24 °C) and pH 6.9-8.5 (optimum, pH 7.8). BP52GT tolerated the addition of up to 1 % NaCl to the medium. Formate and acetate were the major fermentation products. The major cellular fatty acids were C16 : 0, C16:1n-7t and C18:1n-7t. The genome sequence of the isolate was determined. Its G+C content was 30.7 mol%, and the 72.65 % average nucleotide identity of the BP52GT genome to its closest neighbour with a completely sequenced genome (Erysipelatoclostridium ramosum JCM 1298T) indicated low genomic relatedness. Based on the phenotypic and taxonomic characteristics observed in this study, a novel genus and species Tannockella kyphosi gen. nov., sp. nov. is proposed for isolate BP52GT (=NZRM 4757T=JCM 34692T).


Assuntos
Cifose , Tenericutes , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Firmicutes , Bacilos Gram-Positivos/genética , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Tenericutes/genética
5.
Mol Ecol ; 31(8): 2494-2509, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35152505

RESUMO

Many marine herbivorous fishes harbour diverse microbial communities in the hindgut that can play important roles in host health and nutrition. Kyphosus sydneyanus is a temperate marine herbivorous fish that feeds predominantly on brown seaweeds. We employed 16S rRNA gene amplicon sequencing and gas chromatography to characterize microbial communities and their metabolites in different hindgut regions of six K. sydneyanus. Measurements were confined to three distal sections of the intestine, labelled III, IV and V from anterior to posterior. A total of 625 operational taxonomic units from 20 phyla and 123 genera were obtained. Bacteroidota, Firmicutes and Proteobacteria were the major phyla in mean relative abundance, which varied along the gut. Firmicutes (76%) was the most dominant group in section III, whereas Bacteroidota (69.3%) dominated section V. Total short-chain fatty acid (SCFA) concentration was highest in sections IV and V, confirming active fermentation in these two most distal sections. The abundance of Bacteroidota correlated with propionate concentration in section V, while Firmicutes positively correlated with formate in sections III and IV. Acetate levels were highest in sections IV and V, which correlated with abundance of Bacteroidota. Despite differences in gut microbial community composition, SCFA profiles were consistent between individual fish in the different hindgut regions of K. sydneyanus, although proportions of SCFAs differed among gut sections. These findings demonstrate functional compartmentalization of the hindgut microbial community, highlighting the need for regional sampling when interpreting overall microbiome function. These results support previous work suggesting that hindgut microbiota in marine herbivorous fish are important to nutrition in some host species by converting dietary carbohydrates into metabolically useful SCFAs.


Assuntos
Bactérias , Microbiota , Animais , Bacteroidetes/genética , Ácidos Graxos Voláteis/metabolismo , Fermentação , Firmicutes , Peixes/genética , Microbiota/genética , RNA Ribossômico 16S/genética
6.
Animals (Basel) ; 12(4)2022 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-35203206

RESUMO

The gastrointestinal microbiome has a range of roles in the host, including the production of beneficial fermentation end products such as butyrate, which are typically associated with fermentation of plant fibres. However, domestic cats are obligate carnivores and do not require carbohydrates. It has been hypothesised that in the wild, collagenous parts of prey-the so-called animal-derived fermentable substrates (ADFS) such as tendons and cartilage-may be fermented by the cat's gastrointestinal microbiome. However, little research has been conducted on ADFS in the domestic cat. Faecal inoculum was obtained from domestic cats either consuming a high carbohydrate (protein:fat:carbohydrate ratio of 35:20:28 (% dry matter basis)) or high protein (protein:fat:carbohydrate ratio of 75:19:1 (% dry matter basis)) diet. ADFS (hydrolysed collagen, cat hair, and cartilage) were used in a series of static in vitro digestions and fermentations. Concentrations of organic acids and ammonia were measured after 24 h of fermentation, and the culture community of microbes was characterised. The type of inoculum used affected the fermentation profile produced by the ADFS. Butyrate concentrations were highest when hydrolysed collagen was fermented with high protein inoculum (p < 0.05). In contrast, butyrate was not detectable when hydrolysed collagen was fermented in high carbohydrate inoculum (p < 0.05). The microbiome of the domestic cat may be able to ferment ADFS to provide beneficial concentrations of butyrate.

7.
Food Funct ; 12(17): 8007-8016, 2021 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-34269359

RESUMO

Kiwifruit (KF) fiber, a mixture of soluble and insoluble fibers, elicits mucosal changes in the gastrointestinal tract (GIT). This study aimed to define the nature of these changes in mucosal features throughout the GIT of the growing pig in response to semi-synthetic iso-fiber diets containing cellulose (CEL, low GIT luminal functionality) as the sole fiber source (4.5%), or diets where half of the CEL was replaced by either PSY fiber (PSY husk, high GIT luminal functionality) or KF fiber (consumed as intact fruit). Entire male growing pigs (n = 24, 21 kg bodyweight) received the three diets (n = 8) for 42 d. GIT tissues, digesta, and feces were sampled. The partial replacement of CEL increased (P≤ 0.05) the ileal (KF 22% and PSY 33%) and colonic (PSY 86%) mucus layer thickness, whereas it decreased the rectal crypt depth (KF -26%), and small intestinal (duodenum to ileum) villus length (PSY -17%). The number of duodenal goblet cells was 77% higher (P≤ 0.05) for KF than CEL. Pigs fed the KF-containing diet had greater (P≤ 0.05) apparent ileal organic matter digestibility and apparent total tract organic matter digestibility compared with CEL, but the lowest amount of fermented organic matter in the large intestine. In conclusion, partial substitution of CEL with PSY or KF at a constant, practically-relevant dietary fiber intake, affected several measures of GIT functionality with effects being specific to the added fiber.


Assuntos
Actinidia/metabolismo , Celulose/metabolismo , Trato Gastrointestinal/metabolismo , Muco/metabolismo , Psyllium/metabolismo , Suínos/metabolismo , Ração Animal/análise , Animais , Fibras na Dieta/metabolismo , Digestão , Frutas/metabolismo , Trato Gastrointestinal/crescimento & desenvolvimento , Histologia , Masculino , Suínos/crescimento & desenvolvimento
8.
Metabolites ; 10(3)2020 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-32155792

RESUMO

Our understanding of the human gut microbiome has grown exponentially. Advances in genome sequencing technologies and metagenomics analysis have enabled researchers to study microbial communities and their potential function within the context of a range of human gut related diseases and disorders. However, up until recently, much of this research has focused on characterizing the gut microbiological community structure and understanding its potential through system wide (meta) genomic and transcriptomic-based studies. Thus far, the functional output of these microbiomes, in terms of protein and metabolite expression, and within the broader context of host-gut microbiome interactions, has been limited. Furthermore, these studies highlight our need to address the issues of individual variation, and of samples as proxies. Here we provide a perspective review of the recent literature that focuses on the challenges of exploring the human gut microbiome, with a strong focus on an integrated perspective applied to these themes. In doing so, we contextualize the experimental and technical challenges of undertaking such studies and provide a framework for capitalizing on the breadth of insight such approaches afford. An integrated perspective of the human gut microbiome and the linkages to human health will pave the way forward for delivering against the objectives of precision medicine, which is targeted to specific individuals and addresses the issues and mechanisms in situ.

9.
PLoS One ; 14(5): e0216072, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31042730

RESUMO

Commercial diets high in animal protein and fat are increasingly being developed for pets, however little is understood about the impacts of feeding such diets to domestic cats. The carbohydrate content of these diets is typically low, and dietary fibre is often not included. Dietary fibre is believed to be important in the feline gastrointestinal tract, promoting stool formation and providing a substrate for the hindgut microbiome. Therefore, we aimed to determine the effects of adding plant-based dietary fibre to a high animal protein and fat diet. Twelve domestic short hair cats were fed three complete and balanced diets in a cross-over design for blocks of 21 days: raw meat (Raw), raw meat plus fibre (2%, 'as is' inclusion of inulin and cellulose; Raw+Fibre) and a commercially available Kibble diet. A commercially available canned diet was fed for 21 days as a washout phase. Apparent macronutrient digestibility, faecal output, score, pH, organic acid concentrations and bacteriome profiles were determined. Diet significantly affected all faecal parameters measured. The addition of dietary fibre to the raw meat diet was found to reduce apparent macronutrient digestibility, increase faecal output, pH and score. Thirty one bacterial taxa were significantly affected by diet. Prevotella was found to dominate in the Kibble diet, Clostridium and Fusobacterium in the Raw diet, and Prevotella and a group of unclassified Peptostreptococcaceae in the Raw+Fibre diet. Our results show that diets of different macronutrient proportions can strongly influence the faecal microbiome composition and metabolism, as shown by altered organic acid concentrations and faecal pH, in the domestic cat. The addition of 2% of each fibre to the Raw diet shifted faecal parameters closer to those produced by feeding a Kibble diet. These results provide a basis for further research assessing raw red meat diets to domestic cats.


Assuntos
Ração Animal/análise , Dieta Hiperlipídica/veterinária , Dieta Rica em Proteínas/veterinária , Fenômenos Fisiológicos da Nutrição Animal , Animais , Gatos/metabolismo , Dieta/veterinária , Fibras na Dieta/análise , Fezes/química , Feminino , Masculino , Carne/análise , Microbiota/fisiologia , Carne Vermelha
10.
ISME J ; 13(6): 1437-1456, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30728469

RESUMO

Pectin is abundant in modern day diets, as it comprises the middle lamellae and one-third of the dry carbohydrate weight of fruit and vegetable cell walls. Currently there is no specialized model organism for studying pectin fermentation in the human colon, as our collective understanding is informed by versatile glycan-degrading bacteria rather than by specialist pectin degraders. Here we show that the genome of Monoglobus pectinilyticus possesses a highly specialized glycobiome for pectin degradation, unique amongst Firmicutes known to be in the human gut. Its genome encodes a simple set of metabolic pathways relevant to pectin sugar utilization, and its predicted glycobiome comprises an unusual distribution of carbohydrate-active enzymes (CAZymes) with numerous extracellular methyl/acetyl esterases and pectate lyases. We predict the M. pectinilyticus degradative process is facilitated by cell-surface S-layer homology (SLH) domain-containing proteins, which proteomics analysis shows are differentially expressed in response to pectin. Some of these abundant cell surface proteins of M. pectinilyticus share unique modular organizations rarely observed in human gut bacteria, featuring pectin-specific CAZyme domains and the cell wall-anchoring SLH motifs. We observed M. pectinilyticus degrades various pectins, RG-I, and galactan to produce polysaccharide degradation products (PDPs) which are presumably shared with other inhabitants of the human gut microbiome (HGM). This strain occupies a new ecological niche for a primary degrader specialized in foraging a habitually consumed plant glycan, thereby enriching our understanding of the diverse community profile of the HGM.


Assuntos
Colo/microbiologia , Firmicutes/isolamento & purificação , Firmicutes/metabolismo , Pectinas/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Firmicutes/classificação , Firmicutes/genética , Microbioma Gastrointestinal , Humanos , Polissacarídeo-Liases/genética , Polissacarídeo-Liases/metabolismo , Proteômica
11.
AMIA Annu Symp Proc ; 2019: 637-646, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-32308858

RESUMO

With the advent of interoperability standards such as FHIR, SMART, CDS Hooks, and CQL, interoperable clinical decision support (CDS) holds great promise for improving healthcare. In 2018, the Agency for Healthcare Research and Quality (AHRQ)-sponsored Patient-Centered CDS Learning Network (PCCDS LN) chartered a Technical Framework Working Group (TechFWG) to identify barriers, facilitators, and potential solutions for interoperable CDS, with a specific focus on addressing the opioid epidemic. Through an open, multi-stakeholder process that engaged 54 representatives from healthcare, industry, and academia, the TechFWG identified barriers in 6 categories: regulatory environment, data integration, scalability, business case, effective and useful CDS, and care planning and coordination. Facilitators and key recommendations were also identified for overcoming these barriers. The key insights were also extrapolated to CDS-facilitated care improvement outside of the specific opioid use case. If applied broadly, the recommendations should help advance the availability and impact of interoperable CDS delivered at scale.


Assuntos
Analgésicos Opioides/uso terapêutico , Sistemas de Apoio a Decisões Clínicas , Interoperabilidade da Informação em Saúde , Manejo da Dor , Assistência Centrada no Paciente , Tomada de Decisões , Regulamentação Governamental , Humanos , Epidemia de Opioides , Transtornos Relacionados ao Uso de Opioides/terapia , Estados Unidos/epidemiologia , United States Agency for Healthcare Research and Quality
12.
J Am Med Inform Assoc ; 25(12): 1657-1668, 2018 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-30371862

RESUMO

This White Paper presents the foundational domains with examples of key aspects of competencies (knowledge, skills, and attitudes) that are intended for curriculum development and accreditation quality assessment for graduate (master's level) education in applied health informatics. Through a deliberative process, the AMIA Accreditation Committee refined the work of a task force of the Health Informatics Accreditation Council, establishing 10 foundational domains with accompanying example statements of knowledge, skills, and attitudes that are components of competencies by which graduates from applied health informatics programs can be assessed for competence at the time of graduation. The AMIA Accreditation Committee developed the domains for application across all the subdisciplines represented by AMIA, ranging from translational bioinformatics to clinical and public health informatics, spanning the spectrum from molecular to population levels of health and biomedicine. This document will be periodically updated, as part of the responsibility of the AMIA Accreditation Committee, through continued study, education, and surveys of market trends.


Assuntos
Acreditação , Educação de Pós-Graduação/normas , Informática Médica/educação , Competência Profissional , Currículo , Política Organizacional , Sociedades Médicas , Estados Unidos
13.
Front Microbiol ; 9: 1231, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29971046

RESUMO

In humans, aging is associated with changes in the gastrointestinal microbiota; these changes may contribute to the age-related increase in incidence of many chronic diseases, including Type 2 diabetes. The life expectancies of cats are increasing, and they are also exhibiting the same types of diseases. While there are some studies investigating the impacts of diets on gastrointestinal microbiota in young cats, the impacts of aging in older cats has not been explored. We followed a cohort of related kittens, maintained on two commercial diets (kibbled and canned) from weaning (8 weeks) to 5 years of age (260 weeks). We hypothesized that the long-term feeding of specific diet formats would (a) lead to microbial composition changes due to aging, (b) impact body composition, and (c) affect insulin sensitivity in the aging cat. We observed that both diet and age affected fecal microbial composition, and while age correlated with changes in body composition, diet had no effect on body composition. Similarly insulin sensitivity was not affected by age nor diet. 16S rRNA sequencing found unclassified Peptostreptococcaceae were prominent across all ages averaging 21.3% of gene sequence reads and were higher in cats fed canned diets (average of 25.7% of gene sequence reads, vs. 17.0% for kibble-fed cats). Age-related effects on body composition and insulin sensitivity may become apparent as the cats grow older; this study will continue to assess these parameters.

14.
Nutrients ; 10(2)2018 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-29415499

RESUMO

Resistant starch (RS) is the digestion resistant fraction of complex polysaccharide starch. By reaching the large bowel, RS can function as a prebiotic carbohydrate, i.e., it can shape the structure and activity of bowel bacterial communities towards a profile that confers health benefits. However, knowledge about the fate of RS in complex intestinal communities and the microbial members involved in its degradation is limited. In this study, 16S ribosomal RNA (rRNA)-based stable isotope probing (RNA-SIP) was used to identify mouse bowel bacteria involved in the assimilation of RS or its derivatives directly in their natural gut habitat. Stable-isotope [U13C]-labeled native potato starch was administrated to mice, and caecal contents were collected before 0 h and 2 h and 4 h after administration. 'Heavy', isotope-labeled [13C]RNA species, presumably derived from bacteria that have metabolized the labeled starch, were separated from 'light', unlabeled [12C]RNA species by fractionation of isolated total RNA in isopycnic-density gradients. Inspection of different density gradients showed a continuous increase in 'heavy' 16S rRNA in caecal samples over the course of the experiment. Sequencing analyses of unlabeled and labeled 16S amplicons particularly suggested a group of unclassified Clostridiales, Dorea, and a few other taxa (Bacteroides, Turicibacter) to be most actively involved in starch assimilation in vivo. In addition, metabolic product analyses revealed that the predominant 13C-labeled short chain fatty acid (SCFA) in caecal contents produced from the [U13C] starch was butyrate. For the first time, this study provides insights into the metabolic transformation of RS by intestinal bacterial communities directly within a gut ecosystem, which will finally help to better understand its prebiotic potential and possible applications in human health.


Assuntos
Bactérias/metabolismo , Ceco/microbiologia , Microbioma Gastrointestinal/fisiologia , RNA Bacteriano/genética , Amido/metabolismo , Animais , Bactérias/genética , Feminino , Masculino , Camundongos , RNA Ribossômico 16S/genética , Distribuição Aleatória , Organismos Livres de Patógenos Específicos
15.
Int J Syst Evol Microbiol ; 67(12): 4992-4998, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29039307

RESUMO

A novel anaerobic pectinolytic bacterium (strain 14T) was isolated from human faeces. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 14T belonged to the family Ruminococcaceae, but was located separately from known clostridial clusters within the taxon. The closest cultured relative of strain 14T was Acetivibrio cellulolyticus (89.7 % sequence similarity). Strain 14T shared ~99 % sequence similarity with cloned 16S rRNA gene sequences from uncultured bacteria derived from the human gut. Cells were Gram-stain-positive, non-motile cocci approximately 0.6 µm in diameter. Strain 14T fermented pectins from citrus peel, apple, and kiwifruit as well as carbohydrates that are constituents of pectins and hemicellulose, such as galacturonic acid, xylose, and arabinose. TEM images of strain 14T, cultured in association with plant tissues, suggested extracellular fibrolytic activity associated with the bacterial cells, forming zones of degradation in the pectin-rich regions of middle lamella. Phylogenetic and phenotypic analysis supported the differentiation of strain 14T as a novel genus in the family Ruminococcaceae. The name Monoglobus pectinilyticus gen. nov., sp. nov. is proposed; the type strain is 14T (JCM 31914T=DSM 104782T).


Assuntos
Clostridiales/classificação , Fezes/microbiologia , Pectinas/metabolismo , Filogenia , Adulto , Técnicas de Tipagem Bacteriana , Composição de Bases , Clostridiales/genética , Clostridiales/isolamento & purificação , DNA Bacteriano/genética , Feminino , Humanos , Nova Zelândia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
16.
Front Microbiol ; 8: 1331, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28790981

RESUMO

The impact of the intestinal microbiota on human health is becoming increasingly appreciated in recent years. In consequence, and fueled by major technological advances, the composition of the intestinal microbiota in health and disease has been intensively studied by high throughput sequencing approaches. Observations linking dysbiosis of the intestinal microbiota with a number of serious medical conditions including chronic inflammatory disorders and allergic diseases suggest that restoration of the composition and activity of the intestinal microbiota may be a treatment option at least for some of these diseases. One possibility to shape the intestinal microbiota is the administration of prebiotic carbohydrates such as resistant starch (RS). In the present study, we aim at establishing RNA-based stable isotope probing (RNA-SIP) to identify bacterial populations that are involved in the assimilation of RS using anaerobic in vitro fermentation of murine fecal material with stable [U13C] isotope-labeled potato starch. Total RNA from these incubations was extracted, processed by gradient ultracentrifugation and fractionated by density. 16S rRNA gene sequences were amplified from reverse transcribed RNA of high and low density fractions suspected to contain labeled and unlabeled RNA, respectively. Phylogenetic analysis of the obtained sequences revealed a distinct subset of the intestinal microbiota involved in starch metabolism. The results suggest Bacteroidetes, in particular genera affiliated with Prevotellaceae, as well as members of the Ruminococcacea family to be primary assimilators of resistant starch due to a significantly higher relative abundance in higher density fractions in RNA samples isolated after 2 h of incubation. Using high performance liquid chromatography coupled to isotope ratio mass spectrometry (HPLC-IRMS) analysis, some stable isotope label was recovered from acetate, propionate and butyrate. Here, we demonstrate the suitability of RNA-SIP to link specific groups of microorganisms with fermentation of a specific substrate. The application of RNA-SIP in future in vivo studies will help to better understand the mechanisms behind functionality of a prebiotic carbohydrate and its impact on an intestinal ecosystem with potential implications for human health.

17.
Biomed Res Int ; 2017: 1829685, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28299315

RESUMO

RNA-based stable isotope probing (RNA-SIP) and metabolic profiling were used to detect actively glucose-consuming bacteria in a complex microbial community obtained from a murine model system. A faeces-derived microbiota was incubated under anaerobic conditions for 0, 2, and 4 h with 40 mM [U13C]glucose. Isopycnic density gradient ultracentrifugation and fractionation of isolated RNA into labeled and unlabeled fractions followed by 16S rRNA sequencing showed a quick adaptation of the bacterial community in response to the added sugar, which was dominated by unclassified Lachnospiraceae species. Inspection of distinct fractions of isotope-labeled RNA revealed Allobaculum spp. as particularly active glucose utilizers in the system, as the corresponding RNA showed significantly higher proportions among the labeled RNA. With time, the labeled sugar was used by a wider spectrum of faecal bacteria. Metabolic profiling indicated rapid fermentation of [U13C]glucose, with lactate, acetate, and propionate being the principal 13C-labeled fermentation products, and suggested that "cross-feeding" occurred in the system. RNA-SIP combined with metabolic profiling of 13C-labeled products allowed insights into the microbial assimilation of a general model substrate, demonstrating the appropriateness of this technology to study assimilation processes of nutritionally more relevant substrates, for example, prebiotic carbohydrates, in the gut microbiota of mice as a model system.


Assuntos
Firmicutes/metabolismo , Microbioma Gastrointestinal , Glucose/metabolismo , RNA Bacteriano/química , Animais , Isótopos de Carbono/química , Cromatografia Líquida de Alta Pressão , DNA Complementar/metabolismo , Fezes/microbiologia , Fermentação , Firmicutes/genética , Marcação por Isótopo , Camundongos , Camundongos Endogâmicos C57BL , Filogenia , Análise de Componente Principal , RNA Bacteriano/genética , RNA Ribossômico 16S/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA
18.
Food Funct ; 7(4): 1825-32, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26778667

RESUMO

The fate of stable-isotope (13)C labelled and non-labelled inulin catabolism by the gut microbiota was assessed in a healthy rat model. Sprague-Dawley male rats were randomly assigned to diets containing either cellulose or inulin, and were fed these diets for 3 days. On day (d) 4, rats allocated to the inulin diet received (13)C-labelled inulin. The rats were then fed the respective non-labelled diets (cellulose or inulin) until sampling (d4, d5, d6, d7, d10 and d11). Post feeding of (13)C-labelled substrate, breath analysis showed that (13)C-inulin cleared from the host within a period of 36 hours. Faecal (13)C demonstrated the clearance of inulin from gut with a (13)C excess reaching maximum at 24 hours (d5) and then declining gradually. There were greater variations in caecal organic acid concentrations from d4 to d6, with higher concentrations of acetic, butyric and propionic acids observed in the rats fed inulin compared to those fed cellulose. Inulin influenced caecal microbial glycosidase activity, increased colon crypt depth, and decreased the faecal output and polysaccharide content compared to the cellulose diet. In summary, the presence of inulin in the diet positively influenced large bowel microbial fermentation.


Assuntos
Bactérias/metabolismo , Ceco/metabolismo , Intestino Grosso/metabolismo , Inulina/metabolismo , Animais , Isótopos de Carbono/química , Isótopos de Carbono/metabolismo , Ceco/microbiologia , Carboidratos da Dieta/análise , Carboidratos da Dieta/metabolismo , Fezes/química , Fermentação , Microbioma Gastrointestinal , Intestino Grosso/microbiologia , Inulina/química , Marcação por Isótopo , Masculino , Ratos , Ratos Sprague-Dawley
19.
Nutr Res ; 35(1): 23-34, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25530011

RESUMO

This study examined the effect of a Boysenberry beverage (750 mg polyphenols), an apple fiber beverage (7.5 g dietary fiber), and a Boysenberry plus apple fiber beverage (750 mg polyphenols plus 7.5 g dietary fiber) on gut health. Twenty-five individuals completed the study. The study was a placebo-controlled crossover study, where every individual consumed 1 of the 4 treatments in turn. Each treatment phase was 4-week long and was followed by a 2-week washout period. The trial beverages were 350 g taken in 2 doses every day (ie, 175 mL taken twice daily). The hypothesis for the study was that the combination of polyphenols and fiber would have a greater benefit on gut health than the placebo product or the fiber or polyphenols on their own. There were no differences in fecal levels of total bacteria, Bacteroides-Prevotella-Porphyromonas group, Bifidobacteriumspecies, Clostridium perfringens, or Lactobacillus species among any of the treatment groups. Fecal short chain fatty acid concentrations did not vary among treatment groups, although prostaglandin E2 concentrations were higher after consumption of the Boysenberry juice beverage. No significant differences were found in quantitative measures of gut health between the Boysenberry juice beverage, the apple fiber beverage, the Boysenberry juice plus apple fiber beverage, and the placebo beverage.


Assuntos
Bebidas/análise , Fibras na Dieta/administração & dosagem , Ácidos Graxos Voláteis/análise , Fezes/microbiologia , Frutas/química , Polifenóis/administração & dosagem , Adulto , Bacteroides/isolamento & purificação , Bifidobacterium/isolamento & purificação , Índice de Massa Corporal , Peso Corporal , Clostridium perfringens/isolamento & purificação , Estudos Cross-Over , DNA Bacteriano/isolamento & purificação , Dinoprostona/análise , Fezes/química , Feminino , Humanos , Imunoglobulina A/análise , Lactobacillus/isolamento & purificação , Masculino , Malus/química , Pessoa de Meia-Idade , Cooperação do Paciente , Porphyromonas/isolamento & purificação , Prevotella/isolamento & purificação
20.
Adv Food Nutr Res ; 68: 205-17, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23394989

RESUMO

It is becoming clear that the ecology and functionality of the human gut microbiota are extremely diverse and complex. The microbiota have coevolved with us metabolically to live symbiotically and to share the workload of extracting nutrients and energy from the diet. It is also clear that a diet rich in fruit, vegetables, and whole grain cereals is good for general health and gut health and that this is due partly to the phytochemicals and partly to the nondigestible carbohydrates (or dietary fiber) that are present in plants. Kiwifruit contain polyphenolics and nondigestible carbohydrates in the form of pectic, hemicellulosic, and cellulosic polysaccharides, all of which can be degraded by various members of the gut microbiota and result in beneficial effects. This chapter summarizes how kiwifruit act to modify the colonic microbiota and the resultant beneficial effects on human health.


Assuntos
Actinidia/química , Colo/microbiologia , Carboidratos/farmacologia , Fermentação , Humanos , Polifenóis/farmacologia
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