Butyrate is formed in the gut during bacterial fermentation of dietary fiber and is attributed numerous beneficial effects on the host metabolism. We aimed to develop a method for the assessment of functional capacity of gut microbiota butyrate synthesis based on the qPCR quantification of bacterial gene coding butyryl-CoA:acetate CoA-transferase, the key enzyme of butyrate synthesis. In silico, we identified bacteria possessing but gene among human gut microbiota by searching but coding sequences in available databases. We designed and validated six sets of degenerate primers covering all selected bacteria, based on their phylogenetic nearness and sequence similarity, and developed a method for gene abundance normalization in human fecal DNA. We determined but gene abundance in fecal DNA of subjects with opposing dietary patterns and metabolic phenotypes-lean vegans (VG) and healthy obese omnivores (OB) with known fecal microbiota and metabolome composition. We found higher but gene copy number in VG compared with OB, in line with higher fecal butyrate content in VG group. We further found a positive correlation between the relative abundance of target bacterial genera identified by next-generation sequencing and groups of but gene-containing bacteria determined by specific primers. In conclusion, this approach represents a simple and feasible tool for estimation of microbial functional capacity.
- MeSH
- bakteriální geny * MeSH
- butyráty metabolismus MeSH
- DNA bakterií genetika MeSH
- dospělí MeSH
- feces mikrobiologie MeSH
- fenotyp MeSH
- fylogeneze MeSH
- genová dávka MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- neparametrická statistika MeSH
- obezita mikrobiologie MeSH
- polymerázová řetězová reakce * MeSH
- reprodukovatelnost výsledků MeSH
- střevní mikroflóra genetika MeSH
- vegani MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Characterization of metabolites and microbiota composition from human stool provides powerful insight into the molecular phenotypic difference between subjects with normal weight and those with overweight/obesity. The aim of this study was to identify potential metabolic and bacterial signatures from stool that distinguish the overweight/obesity state in children/adolescents. Using 1H NMR spectral analysis and 16S rRNA gene profiling, the fecal metabolic profile and bacterial composition from 52 children aged 7 to 16 was evaluated. The children were classified into three groups (16 with normal-weight, 17 with overweight, 19 with obesity). The metabolomic analysis identified four metabolites that were significantly different (p < 0.05) among the study groups based on one-way ANOVA testing: arabinose, butyrate, galactose, and trimethylamine. Significantly different (p < 0.01) genus-level taxa based on edgeR differential abundance tests were genus Escherichia and Tyzzerella subgroup 3. No significant difference in alpha-diversity was detected among the three study groups, and no significant correlations were found between the significant taxa and metabolites. The findings support the hypothesis of increased energy harvest in obesity by human gut bacteria through the growing observation of increased fecal butyrate in children with overweight/obesity, as well as an increase of certain monosaccharides in the stool. Also supported is the increase of trimethylamine as an indicator of an unhealthy state.
- MeSH
- Bacteria genetika MeSH
- dítě MeSH
- feces mikrobiologie MeSH
- lidé MeSH
- metabolom MeSH
- metabolomika MeSH
- mladiství MeSH
- obezita dětí a dospívajících metabolismus mikrobiologie MeSH
- obezita metabolismus mikrobiologie MeSH
- protonová magnetická rezonanční spektroskopie MeSH
- RNA ribozomální 16S genetika MeSH
- střevní mikroflóra genetika MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Dietary supplementation with calanus oil, a novel wax ester-rich marine oil, has been shown to reduce adiposity in high-fat diet (HFD)-induced obese mice. Current evidence suggests that obesity and its comorbidities are intrinsically linked with unfavorable changes in the intestinal microbiome. Thus, in line with its antiobesity effect, we hypothesized that dietary supplementation with calanus oil should counteract the obesity-related deleterious changes in the gut microbiota. Seven-week-old female C57bl/6J mice received an HFD for 12 weeks to induce obesity followed by 8-week supplementation with 2% calanus oil. For comparative reasons, another group of mice was treated with exenatide, an antiobesogenic glucagon-like peptide-1 receptor agonist. Mice fed normal chow diet or nonsupplemented HFD for 20 weeks served as lean and obese controls, respectively. 16S rRNA gene sequencing was performed on fecal samples from the colon. HFD increased the abundance of the Lactococcus and Leuconostoc genera relative to normal chow diet, whereas abundances of Allobaculum and Oscillospira were decreased. Supplementation with calanus oil led to an apparent overrepresentation of Lactobacillus and Streptococcus and underrepresentation of Bilophila. Exenatide prevented the HFD-induced increase in Lactococcus and caused a decrease in the abundance of Streptococcus compared to the HFD group. Thus, HFD altered the gut microbiota composition in an unhealthy direction by increasing the abundance of proinflammatory genera while reducing those considered health-promoting. These obesity-induced changes were antagonized by both calanus oil and exenatide.
- MeSH
- Bacteria klasifikace genetika růst a vývoj MeSH
- dieta s vysokým obsahem tuků * MeSH
- dietní tuky nenasycené aplikace a dávkování MeSH
- exenatid farmakologie MeSH
- feces mikrobiologie MeSH
- hmotnostní přírůstek MeSH
- kolon mikrobiologie MeSH
- látky proti obezitě farmakologie MeSH
- metagenom MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- obezita mikrobiologie patofyziologie terapie MeSH
- oleje aplikace a dávkování MeSH
- potravní doplňky * MeSH
- střevní mikroflóra * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Gut microbiome lives in the intestinal tract of animals and plays an important role in almost all life processes. Gut microbiome balance is beneficial to health, and imbalance leads to many diseases, one of which is obesity epidemic. However, gut microbiome is also influenced by host hormone, and different gut microbiome composition is observed between the sexes. Here, we studied whether castrated male Guizhou minipigs with obesity own the same gut microbiome composition and microbial function predictions with those in obese females. We sequenced the hypervariable regions V3 to V4 of bacterial 16s rRNA of fecal samples collected from our study subjects. We observed that the operational taxonomic units were small, which suggested that the abundance of gut microbiome may be influenced by low genetic diversity of host. Our results also suggested that the castrated male has different gut microbial composition compared to the obese female. An increasing Firmicutes/Bacteroidetes ratio was observed in both castrated male and obese female groups, which suggested that the main adipogenic gut microorganism in obese Guizhou minipigs in our studies is the same with that in other obese mammals. However, we also observed that there were function prediction differences of obese Guizhou minipigs between female and castrated male, which suggested that the influence of gut microbiome on obesity between them is different.
- MeSH
- Bacteria klasifikace genetika izolace a purifikace MeSH
- DNA bakterií genetika MeSH
- feces mikrobiologie MeSH
- fylogeneze MeSH
- kastrace * MeSH
- miniaturní prasata MeSH
- modely nemocí na zvířatech MeSH
- obezita mikrobiologie MeSH
- prasata MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- střevní mikroflóra * genetika MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
Obesity is linked to a wide range of serious illnesses. In addition to the important impact on the health of the individual, obesity also has a substantial impact on the economy. Disruption of physiological day-night cycles could contribute to the increased incidence of obesity. According to the American National Sleep Federation, the percentage of the people who reported a sleep duration of six hours or less increased from 12 to 37 % over ten years. Insufficient sleep leads not only to an increase of the total calorie intake but changes the meal preference in favor of palatable foods and meals with high carbohydrate content. A decrease of leptin and increase of ghrelin levels caused by sleep deficiency can also play a role. In addition to the higher caloric intake, the timing of food consumption should be taken into account. The same meal eaten during the night versus the day is associated with increased postprandial glucose and triglyceride levels. The gut microbiome has also been recently understood as an endocrine system, with links between the gut microbiome and circadian rhythm changes possibly influencing increased obesity.
- MeSH
- cirkadiánní rytmus fyziologie MeSH
- energetický příjem fyziologie MeSH
- lidé MeSH
- obezita komplikace metabolismus mikrobiologie MeSH
- postprandiální období fyziologie MeSH
- spánková deprivace komplikace metabolismus mikrobiologie MeSH
- střevní mikroflóra fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- MeSH
- dospělí MeSH
- gastrointestinální nemoci prevence a kontrola MeSH
- infekce dýchací soustavy prevence a kontrola MeSH
- kojenec MeSH
- kojení * MeSH
- lidé MeSH
- Limosilactobacillus fermentum * MeSH
- mastitida farmakoterapie MeSH
- mateřské mléko mikrobiologie MeSH
- obezita etiologie mikrobiologie MeSH
- probiotika * terapeutické užití MeSH
- střevní mikroflóra MeSH
- Check Tag
- dospělí MeSH
- kojenec MeSH
- lidé MeSH
- ženské pohlaví MeSH
- MeSH
- ateroskleróza etiologie MeSH
- Bacteria klasifikace růst a vývoj MeSH
- diabetes mellitus etiologie MeSH
- gastrointestinální trakt mikrobiologie MeSH
- lidé MeSH
- myši MeSH
- obezita * etiologie mikrobiologie parazitologie prevence a kontrola MeSH
- střevní sliznice * fyziologie mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
Gastrointestinální ekosystém je vysoce integrovaný komplex, který zahrnuje střevní mikrobiom, střevní slizniční bariéru a slizniční imunitní systém. Tento komplex zajišťuje látkovou přeměnu, výživu a regulaci slizniční a systémové imunity hostitele. Probiotika jsou živé mikroorganismy převážně lidského původu, které při podání v přiměřeném množství příznivě ovlivňují zdravotní stav hostitele. V úzké spolupráci s pre- biotiky se podílejí na udržování rovnováhy jednotlivých složek gastrointestinálního ekosystému, jejichž poruchy se uplatňují v patogenezi řady chorob včetně masových civilizačních nemocí. Modifikace střevní- ho mikrobiomu racionálním podáváním antibiotik, probiotik a prebiotik umožní u některých chorob nové terapeutické postupy.
Gastrointestinal ecosystem is a highly integrated complex including intestinal microbiome, intestinal mucosal barrier, and mucosal immune system. The complex retains the metabolism, nutrition, and regulation of the mucosal and systemic immunity of the host. Probiotics are living microorganisms predominantly of human origin which applied in adequate amounts may confer a benefit on the host. Probiotics in close cooperation with prebiotics maintain the balance of individual components of the gastrointestinal ecosystem. Their disturbances participate in the pathogenesis of different diseases including civilization diseases of mass occurrence. Modification of the intestinal microbiome using rational application of antibiotics, probiotics, and prebiotics will enable to develop new therapeutic approaches.
- Klíčová slova
- metabolická endotoxémie,
- MeSH
- antibakteriální látky aplikace a dávkování škodlivé účinky MeSH
- diabetes mellitus 2. typu MeSH
- dieta s omezením tuků využití MeSH
- inzulinová rezistence MeSH
- lidé MeSH
- obezita dietoterapie enzymologie mikrobiologie MeSH
- prebiotika mikrobiologie využití MeSH
- probiotika farmakologie terapeutické užití MeSH
- střevní sliznice mikrobiologie MeSH
- synbiotika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- adipozita fyziologie MeSH
- alkalická fosfatasa metabolismus MeSH
- Bacteroides izolace a purifikace MeSH
- Enterococcus izolace a purifikace MeSH
- financování organizované MeSH
- krysa rodu rattus MeSH
- Lactobacillus izolace a purifikace MeSH
- metagenom MeSH
- obezita mikrobiologie MeSH
- odstavení MeSH
- potkani Sprague-Dawley MeSH
- Prevotella izolace a purifikace MeSH
- riziko MeSH
- střeva mikrobiologie MeSH
- tělesná hmotnost MeSH
- velikost vrhu fyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
The influence was evaluated of post-weaning normal nutrition and over-nutrition upon the development of the intestinal microbiota, the alkaline phosphatase activity (AP) and occurrence of obesity in male Sprague-Dawley rats (from days 21 to 40 the control rats were submitted to ad libitum intake of a standard laboratory diet whereas overfed rats received the same diet supplemented with milk-based high fat liquid diet). The jejunal numbers of two dominant divisions of bacteria, i.e. Firmicutes (Lactobacillus/ Enterococcus--LAB) and the Bacteroidetes (Bacteroides/Prevotella--BAC), were determined using the fluorescent in situ hybridization (FISH) method, and the jejunal AP activity was assayed histochemically. On day 40, the overfed rats in comparison with control animals displayed increased adiposity accompanied by enhanced AP activity, abundance of LAB, lower amounts of BAC and, thereafter, higher LAB/BAC ratio (L/B). The numbers of LAB and L/B index positively correlated with body fat, energy intake and AP activity, whereas numbers of BAC showed an opposite tendency. These results revealed the significance of nutritional imprint upon the post-weaning development of intestinal microbial and functional axis and contribute to better understanding of their involvement in energy-balance control and in adverse and/or positive regulation of adiposity.
- MeSH
- alkalická fosfatasa metabolismus MeSH
- Bacteroidetes izolace a purifikace klasifikace MeSH
- biodiverzita MeSH
- dieta MeSH
- financování organizované MeSH
- grampozitivní bakterie izolace a purifikace klasifikace MeSH
- histocytochemie MeSH
- hybridizace in situ fluorescenční MeSH
- jejunum enzymologie mikrobiologie MeSH
- krysa rodu rattus MeSH
- obezita mikrobiologie MeSH
- potkani Sprague-Dawley MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH