Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 16 de 16
Filtrar
Más filtros










Base de datos
Intervalo de año de publicación
1.
Nat Commun ; 14(1): 7770, 2023 Nov 27.
Artículo en Inglés | MEDLINE | ID: mdl-38012160

RESUMEN

The transgenic 116C-NOD mouse strain exhibits a prevalent Th17 phenotype, and reduced type 1 diabetes (T1D) compared to non-obese diabetic (NOD) mice. A cohousing experiment between both models revealed lower T1D incidence in NOD mice cohoused with 116C-NOD, associated with gut microbiota changes, reduced intestinal permeability, shifts in T and B cell subsets, and a transition from Th1 to Th17 responses. Distinct gut bacterial signatures were linked to T1D in each group. Using a RAG-2-/- genetic background, we found that T cell alterations promoted segmented filamentous bacteria proliferation in young NOD and 116C-NOD, as well as in immunodeficient NOD.RAG-2-/- and 116C-NOD.RAG-2-/- mice across all ages. Bifidobacterium colonization depended on lymphocytes and thrived in a non-diabetogenic environment. Additionally, 116C-NOD B cells in 116C-NOD.RAG-2-/- mice enriched the gut microbiota in Adlercreutzia and reduced intestinal permeability. Collectively, these results indicate reciprocal modulation between gut microbiota and the immune system in rodent T1D models.


Asunto(s)
Subgrupos de Linfocitos B , Diabetes Mellitus Tipo 1 , Microbioma Gastrointestinal , Ratones , Animales , Diabetes Mellitus Tipo 1/genética , Ratones Endogámicos NOD , Microbioma Gastrointestinal/genética , Linfocitos B
2.
Gut Microbes ; 15(1): 2205425, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37131291

RESUMEN

Emerging evidence implicates microbial proteolytic activity in ulcerative colitis (UC), but whether it also plays a role in Crohn's disease (CD) remains unclear. We investigated the effects of colonizing adult and neonatal germ-free C57BL/6 mice with CD microbiota, selected based on high (CD-HPA) or low fecal proteolytic activity (CD-LPA), or microbiota from healthy controls with LPA (HC-LPA) or HPA (HC-HPA). We then investigated colitogenic mechanisms in gnotobiotic C57BL/6, and in mice with impaired Nucleotide-binding Oligomerization Domain-2 (NOD2) and Protease-Activated Receptor 2 (PAR2) cleavage resistant mice (Nod2-/-; R38E-PAR2 respectively). At sacrifice, total fecal proteolytic, elastolytic, and mucolytic activity were analyzed. Microbial community and predicted function were assessed by 16S rRNA gene sequencing and PICRUSt2. Immune function and colonic injury were investigated by inflammatory gene expression (NanoString) and histology. Colonization with HC-LPA or CD-LPA lowered baseline fecal proteolytic activity in germ-free mice, which was paralleled by lower acute inflammatory cell infiltrate. CD-HPA further increased proteolytic activity compared with germ-free mice. CD-HPA mice had lower alpha diversity, distinct microbial profiles and higher fecal proteolytic activity compared with CD-LPA. C57BL/6 and Nod2-/- mice, but not R38E-PAR2, colonized with CD-HPA had higher colitis severity than those colonized with CD-LPA. Our results indicate that CD proteolytic microbiota is proinflammatory, increasing colitis severity through a PAR2 pathway.


Asunto(s)
Colitis Ulcerosa , Colitis , Enfermedad de Crohn , Microbioma Gastrointestinal , Microbiota , Animales , Ratones , Ratones Endogámicos C57BL , Receptor PAR-2/genética , ARN Ribosómico 16S/genética , Inflamación , Serina Proteasas
3.
Nat Aging ; 2(10): 941-955, 2022 10.
Artículo en Inglés | MEDLINE | ID: mdl-36398033

RESUMEN

Older adults represent a vulnerable population with elevated risk for numerous morbidities. To explore the association of the microbiome with aging and age-related susceptibilities including frailty and infectious disease risk, we conducted a longitudinal study of the skin, oral, and gut microbiota in 47 community- or skilled nursing facility-dwelling older adults vs. younger adults. We found that microbiome changes were not associated with chronological age so much as frailty: we identified prominent changes in microbiome features associated with susceptibility to pathogen colonization and disease risk, including diversity, stability, heterogeneity, and biogeographic determinism, which were moreover associated with a loss of Cutibacterium (C.) acnes in the skin microbiome. Strikingly, the skin microbiota were also the primary reservoir for antimicrobial resistance, clinically important pathobionts, and nosocomial strains, suggesting a potential role particularly for the skin microbiome in disease risk and dissemination of multidrug resistant pathogens.


Asunto(s)
Fragilidad , Microbioma Gastrointestinal , Infecciones , Microbiota , Humanos , Anciano , Fragilidad/epidemiología , Estudios Longitudinales , Susceptibilidad a Enfermedades/microbiología
4.
Gastroenterology ; 160(5): 1532-1545, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33310084

RESUMEN

BACKGROUND & AIMS: Altered gut microbiota composition and function have been associated with inflammatory bowel diseases, including ulcerative colitis (UC), but the causality and mechanisms remain unknown. METHODS: We applied 16S ribosomal RNA gene sequencing, shotgun metagenomic sequencing, in vitro functional assays, and gnotobiotic colonizations to define the microbial composition and function in fecal samples obtained from a cohort of healthy individuals at risk for inflammatory bowel diseases (pre-UC) who later developed UC (post-UC) and matched healthy control individuals (HCs). RESULTS: Microbiota composition of post-UC samples was different from HC and pre-UC samples; however, functional analysis showed increased fecal proteolytic and elastase activity before UC onset. Metagenomics identified more than 22,000 gene families that were significantly different between HC, pre-UC, and post-UC samples. Of these, 237 related to proteases and peptidases, suggesting a bacterial component to the pre-UC proteolytic signature. Elastase activity inversely correlated with the relative abundance of Adlercreutzia and other potentially beneficial taxa and directly correlated with known proteolytic taxa, such as Bacteroides vulgatus. High elastase activity was confirmed in Bacteroides isolates from fecal samples. The bacterial contribution and functional significance of the proteolytic signature were investigated in germ-free adult mice and in dams colonized with HC, pre-UC, or post-UC microbiota. Mice colonized with or born from pre-UC-colonized dams developed higher fecal proteolytic activity and an inflammatory immune tone compared with HC-colonized mice. CONCLUSIONS: We have identified increased fecal proteolytic activity that precedes the clinical diagnosis of UC and associates with gut microbiota changes. This proteolytic signature may constitute a noninvasive biomarker of inflammation to monitor at-risk populations that can be targeted therapeutically with antiproteases.


Asunto(s)
Bacterias/enzimología , Proteínas Bacterianas/metabolismo , Colitis Ulcerosa/microbiología , Heces/microbiología , Microbioma Gastrointestinal , Péptido Hidrolasas/metabolismo , Adolescente , Adulto , Animales , Bacterias/efectos de los fármacos , Bacterias/genética , Proteínas Bacterianas/genética , Biomarcadores/metabolismo , Estudios de Casos y Controles , Niño , Colitis Ulcerosa/diagnóstico , Colitis Ulcerosa/tratamiento farmacológico , Modelos Animales de Enfermedad , Trasplante de Microbiota Fecal , Femenino , Microbioma Gastrointestinal/efectos de los fármacos , Vida Libre de Gérmenes , Humanos , Masculino , Metagenoma , Metagenómica , Ratones Endogámicos C57BL , Péptido Hidrolasas/genética , Valor Predictivo de las Pruebas , Estudios Prospectivos , Inhibidores de Proteasas/uso terapéutico , Proteolisis , Reproducibilidad de los Resultados , Ribotipificación , Adulto Joven
5.
J Crohns Colitis ; 14(11): 1535-1546, 2020 Nov 07.
Artículo en Inglés | MEDLINE | ID: mdl-32333762

RESUMEN

BACKGROUND AND AIMS: Intestinal microbiota dysbiosis is implicated in Crohn's disease [CD] and may play an important role in triggering postoperative disease recurrence [POR]. We prospectively studied faecal and mucosal microbial recolonisation following ileocaecal resection to identify the predictive value of recurrence-related microbiota. METHODS: Mucosal and/or faecal samples from 121 CD patients undergoing ileocaecal resection were collected at predefined time points before and after surgery. Ileal biopsies were collected from 39 healthy controls. POR was defined by a Rutgeerts score ≥i2b. The microbiota was evaluated by 16S rRNA sequencing. Prediction analysis was performed using C5.0 and Random Forest algorithms. RESULTS: The mucosa-associated microbiota in CD patients was characterised by a depletion of butyrate-producing species (false discovery rate [FDR] <0.01) and enrichment of Proteobacteria [FDR = 0.009] and Akkermansia spp. [FDR = 0.02]. Following resection, a mucosal enrichment of Lachnospiraceae [FDR <0.001] was seen in all patients but in POR patients, also Fusobacteriaceae [FDR <0.001] increased compared with baseline. Patients without POR showed a decrease of Streptococcaceae [FDR = 0.003] and Actinomycineae [FDR = 0.06]. The mucosa-associated microbiota profile had good discriminative power to predict POR, and was superior to clinical risk factors. At Month 6, patients experiencing POR had a higher abundance of taxa belonging to Negativicutes [FDR = 0.04] and Fusobacteria [FDR = 0.04] compared with patients without POR. CONCLUSIONS: Microbiota recolonisation after ileocaecal resection is different between recurrence and non-recurrence patients, with Fusobacteria as the most prominent player driving early POR. These bacteria involved in the early recolonisation and POR represent a promising therapeutic strategy in the prevention of disease recurrence.


Asunto(s)
Enfermedad de Crohn , Disbiosis , Fusobacterias/aislamiento & purificación , Microbioma Gastrointestinal/fisiología , Mucosa Intestinal , Complicaciones Posoperatorias , Adulto , Bélgica/epidemiología , Biopsia/métodos , Ciego/cirugía , Enfermedad de Crohn/epidemiología , Enfermedad de Crohn/microbiología , Enfermedad de Crohn/patología , Enfermedad de Crohn/cirugía , Procedimientos Quirúrgicos del Sistema Digestivo/efectos adversos , Procedimientos Quirúrgicos del Sistema Digestivo/métodos , Disbiosis/diagnóstico , Disbiosis/etiología , Disbiosis/microbiología , Heces/química , Heces/microbiología , Femenino , Humanos , Íleon/cirugía , Mucosa Intestinal/microbiología , Mucosa Intestinal/patología , Masculino , Complicaciones Posoperatorias/diagnóstico , Complicaciones Posoperatorias/microbiología , Recurrencia , Resultado del Tratamiento
6.
EBioMedicine ; 51: 102611, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31901867

RESUMEN

BACKGROUND: The remission of Crohn's disease (CD) can be accomplished by faecal microbiota transplantation (FMT). However, this procedure has a low success rate, which could be attributed to mis-communication between recipient intestinal mucosa and donor microbiota. METHODS: Here we used a human explant tissue model and an in vivo mouse model to examine changes in recipient intestinal mucosa upon contact with a faecal suspension (FS) obtained from a healthy donor. CD patients provided resected inflamed and non-inflamed mucosal tissues, whereas control colonic mucosa samples were collected from colorectal cancer patients. For the models, mucosal microbiome composition and tissue response were evaluated. FINDINGS: We show that cytokine release and tissue damage were significantly greater in inflamed compared to non-inflamed CD tissues. Moreover, mucosal samples harbouring an initial low microbial load presented a shift in composition towards that of the FS, an increase in the relative count of Faecalibacterium prausnitzii, and a higher secretion of anti-inflammatory cytokine IL-10 compared to those with a high microbial load. INTERPRETATION: Our results indicate that FMT during active inflammatory disease can compromise treatment outcome. We recommend the stratification of FMT recipients on the basis of tissue microbial load as a strategy to ensure successful colonization. FUNDING: This study was supported by grants from the Instituto de Salud Carlos III/FEDER (PI17/00614), the European Commission: (INCOMED-267128) and PERIS (SLT002/16). K.M. is a postdoctoral fellow and S.V. a senior clinical investigator of the Fund for Scientific Research Flanders, Belgium (FWO-Vlaanderen).


Asunto(s)
Enfermedad de Crohn/microbiología , Trasplante de Microbiota Fecal , Heces/microbiología , Mucosa Intestinal/microbiología , Animales , Enfermedad de Crohn/patología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Humanos , Inflamación/microbiología , Inflamación/patología , Mucosa Intestinal/patología , Ratones , Modelos Biológicos , Reproducibilidad de los Resultados
7.
EBioMedicine ; 48: 630-641, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31628021

RESUMEN

BACKGROUND: Faecal microbiota transplantation (FMT) is a novel potential therapy for inflammatory bowel diseases, but it is poorly characterised. METHODS: We evaluated the performance of the mouse and rat as a pre-clinical model for human microbiota engraftment. We then characterised the effect of a single human stool transfer (HST) on a humanised model of DSS-induced colitis. Colonic and faecal microbial communities were analysed using the 16S rRNA approach and clinical manifestations were assessed in a longitudinal setting. FINDINGS: The microbial community of rats showed greater similarity to that of humans, while the microbiome of mice showed less similarity to that of humans. Moreover, rats captured more human microbial species than mice after a single HST. Using the rat model, we showed that HST compensated faecal dysbiosis by restoring alpha-diversity and by increasing the relative abundance of health-related microbial genera. To some extent, HST also modulated the microbial composition of colonic tissue. These faecal and colonic microbial communities alterations led to a relative restoration of colon length, and a significant decrease in both epithelium damage and disease severity. Remarkably, stopping inflammation by removing DSS before HST caused a faster and greater recovery of both microbiome and clinical manifestation features. INTERPRETATION: Our results indicate that the rat outperforms the mouse as a model for human microbiota engraftment and show that the efficacy of HST can be enhanced when inflammation stimulation is withdrawn. Finally, our findings support a new therapeutic strategy based on the use FMT combined with anti-inflammatory drugs.


Asunto(s)
Colitis/etiología , Colitis/terapia , Trasplante de Microbiota Fecal , Microbioma Gastrointestinal , Interacciones Microbianas , Animales , Biodiversidad , Biomarcadores , Biopsia , Colitis/patología , Modelos Animales de Enfermedad , Trasplante de Microbiota Fecal/métodos , Humanos , Enfermedades Inflamatorias del Intestino/etiología , Enfermedades Inflamatorias del Intestino/patología , Enfermedades Inflamatorias del Intestino/terapia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Mucosa Intestinal/patología , Masculino , Ratones , Ratas , Resultado del Tratamiento
8.
mSystems ; 4(1)2019.
Artículo en Inglés | MEDLINE | ID: mdl-30801032

RESUMEN

Whether the interaction between the gut microbiota and the immune response influences the evolution of cirrhosis is poorly understood. We aimed to investigate modifications of the microbiome and the immune response during the progression of cirrhosis. Rats were treated with carbon tetrachloride (CCl4) to induce cirrhosis. We then assessed microbiome load and composition in stool, ileocecal contents (ICCs), mesenteric lymph nodes (MLNs), blood, and ascitic fluids (AFs) at 6, 8, and 10 weeks or ascites production and measured cytokine production in MLNs and blood. The microbiome of MLN, blood, and AF showed a distinct composition compared to that of stool and ICCs. Betaproteobacteria (Sutterella) were found associated with the appearance of a decompensated state of cirrhosis. Microbial load increased and showed a positive correlation with the relative abundance of pathobionts in the MLN of decompensated rats. Among several genera, Escherichia and "Candidatus Arthromitus" positively correlated with elevated levels of systemic proinflammatory cytokines. "Candidatus Arthromitus," a segmented filamentous bacteria, was detected in ICC, MLN, and AF samples, suggesting a possible translocation from the gut to the AF through the lymphatic system, whereas Escherichia was detected in ICC, MLN, AF, and blood, suggesting a possible translocation from the gut to the AF through the bloodstream. In the present study, we demonstrate that microbiome changes in distinct intestinal sites are associated with microbial shifts in the MLNs as well as an increase in cytokine production, providing further evidence of the role the gut-liver-immunity axis plays in the progression of cirrhosis. IMPORTANCE Cirrhosis severity in patients was previously shown to be associated with progressive changes in the fecal microbiome in a longitudinal setting. Recent evidence shows that bacterial translocation from the gut to the extraintestinal sites could play a major role in poor disease outcome and patient survival. However, the underlying mechanisms involving the microbiota in the disease progression are not well understood. Here, using an animal model of cirrhosis in a longitudinal and multibody sites setting, we showed the presence of a distinct composition of the microbiome in mesenteric lymph nodes, blood, and ascitic fluid compared to that in feces and ileocecal content, suggesting compartmentalization of the gut microbiome. We also demonstrate that microbiome changes in intestinal sites are associated with shifts in specific microbial groups in the mesenteric lymph nodes as well as an increase in systemic cytokine production, linking inflammation to decompensated cirrhosis in the gut-liver-immunity axis.

9.
Hepatology ; 67(4): 1485-1498, 2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29113028

RESUMEN

Portal hypertension (PH) drives most of the clinical complications in chronic liver diseases. However, its progression in nonalcoholic steatohepatitis (NASH) and its association with the intestinal microbiota (IM) have been scarcely studied. Our aim was to investigate the role of the IM in the mechanisms leading to PH in early NASH. The experimental design was divided in two stages. In stage 1, Sprague-Dawley rats were fed for 8 weeks a high-fat, high-glucose/fructose diet (HFGFD) or a control diet/water (CD). Representative rats were selected as IM donors for stage 2. In stage 2, additional HFGFD and CD rats underwent intestinal decontamination, followed by IM transplantation with feces from opposite-diet donors (heterologous transplant) or autologous fecal transplant (as controls), generating four groups: CD-autotransplanted, CD-transplanted, HFGFD-autotransplanted, HFGFD-transplanted. After IM transplantation, the original diet was maintained for 12-14 days until death. HFGFD rats developed obesity, insulin resistance, NASH without fibrosis but with PH, intrahepatic endothelial dysfunction, and IM dysbiosis. In HFGFD rats, transplantation with feces from CD donors caused a significant reduction of PH to levels comparable to CD without significant changes in NASH histology. The reduction in PH was due to a 31% decrease of intrahepatic vascular resistance compared to the HFGFD-autotransplanted group (P < 0.05). This effect occurs through restoration of the sensitivity to insulin of the hepatic protein kinase B-dependent endothelial nitric oxide synthase signaling pathway. CONCLUSION: The IM exerts a direct influence in the development of PH in rats with diet-induced NASH and dysbiosis; PH, insulin resistance, and endothelial dysfunction revert when a healthy IM is restored. (Hepatology 2018;67:1485-1498).


Asunto(s)
Disbiosis/complicaciones , Microbioma Gastrointestinal , Hipertensión Portal/etiología , Enfermedad del Hígado Graso no Alcohólico/complicaciones , Animales , Modelos Animales de Enfermedad , Trasplante de Microbiota Fecal/métodos , Hipertensión Portal/microbiología , Resistencia a la Insulina , Hígado/patología , Masculino , Ratas , Ratas Sprague-Dawley
10.
Sci Rep ; 7: 45712, 2017 04 03.
Artículo en Inglés | MEDLINE | ID: mdl-28368023

RESUMEN

Probiotics can prevent pathological bacterial translocation by modulating intestinal microbiota and improving the gut barrier. The aim was to evaluate the effect of a fermented milk containing Lactobacillus paracasei subsp. paracasei CNCM I-1518 on bacterial translocation in rats with carbon tetrachloride (CCl4)-induced cirrhosis. Sprague-Dawley rats treated with CCl4 were randomized into a probiotic group that received fermented milk containing Lactobacillus paracasei subsp. paracasei CNCM I-1518 in drinking water or a water group that received water only. Laparotomy was performed one week after ascites development. We evaluated bacterial translocation, intestinal microbiota, the intestinal barrier and cytokines in mesenteric lymph nodes and serum. Bacterial translocation decreased and gut dysbiosis improved in the probiotic group compared to the water group. The ileal ß-defensin-1 concentration was higher and ileal malondialdehyde levels were lower in the probiotic group than in water group. There were no differences between groups in serum cytokines but TNF-α levels in mesenteric lymph nodes were lower in the probiotic group than in the water group. Fermented milk containing Lactobacillus paracasei subsp. paracasei CNCM I-1518 decreases bacterial translocation, gut dysbiosis and ileal oxidative damage and increases ileal ß-defensin-1 expression in rats treated with CCl4, suggesting an improvement in the intestinal barrier integrity.


Asunto(s)
Traslocación Bacteriana/fisiología , Microbioma Gastrointestinal/fisiología , Lacticaseibacillus paracasei/fisiología , Leche/microbiología , Animales , Traslocación Bacteriana/efectos de los fármacos , Tetracloruro de Carbono/toxicidad , Disbiosis/metabolismo , Disbiosis/fisiopatología , Disbiosis/prevención & control , Fermentación , Microbioma Gastrointestinal/efectos de los fármacos , Cirrosis Hepática Experimental/inducido químicamente , Cirrosis Hepática Experimental/prevención & control , Masculino , Malondialdehído/metabolismo , Probióticos/farmacología , Ratas Sprague-Dawley , beta-Defensinas/metabolismo
11.
Gut ; 66(5): 813-822, 2017 05.
Artículo en Inglés | MEDLINE | ID: mdl-28179361

RESUMEN

OBJECTIVE: A decade of microbiome studies has linked IBD to an alteration in the gut microbial community of genetically predisposed subjects. However, existing profiles of gut microbiome dysbiosis in adult IBD patients are inconsistent among published studies, and did not allow the identification of microbial signatures for CD and UC. Here, we aimed to compare the faecal microbiome of CD with patients having UC and with non-IBD subjects in a longitudinal study. DESIGN: We analysed a cohort of 2045 non-IBD and IBD faecal samples from four countries (Spain, Belgium, the UK and Germany), applied a 16S rRNA sequencing approach and analysed a total dataset of 115 million sequences. RESULTS: In the Spanish cohort, dysbiosis was found significantly greater in patients with CD than with UC, as shown by a more reduced diversity, a less stable microbial community and eight microbial groups were proposed as a specific microbial signature for CD. Tested against the whole cohort, the signature achieved an overall sensitivity of 80% and a specificity of 94%, 94%, 89% and 91% for the detection of CD versus healthy controls, patients with anorexia, IBS and UC, respectively. CONCLUSIONS: Although UC and CD share many epidemiologic, immunologic, therapeutic and clinical features, our results showed that they are two distinct subtypes of IBD at the microbiome level. For the first time, we are proposing microbiomarkers to discriminate between CD and non-CD independently of geographical regions.


Asunto(s)
Colitis Ulcerosa/microbiología , Enfermedad de Crohn/diagnóstico , Enfermedad de Crohn/microbiología , Disbiosis/microbiología , Heces/microbiología , ARN Bacteriano/análisis , ARN Ribosómico 16S/análisis , Adolescente , Adulto , Anciano , Bélgica , Biomarcadores , Estudios de Casos y Controles , Heces/química , Femenino , Microbioma Gastrointestinal , Alemania , Humanos , Complejo de Antígeno L1 de Leucocito/análisis , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Fumar , España , Reino Unido , Adulto Joven
12.
Sci Rep ; 6: 25001, 2016 04 26.
Artículo en Inglés | MEDLINE | ID: mdl-27112233

RESUMEN

The progression of cirrhosis is associated with alterations in the composition of the gut microbiome. To assess microbial translocation, we compared the serum microbial composition of patients with and without ascites and characterized the ascitic fluid microbiome using 16S rDNA high-throughput sequencing data. A complex and specific microbial community was detected in the serum and ascitic fluid of patients with cirrhosis but barely detectable in the serum of healthy controls. The serum microbiome of patients with ascites presented higher levels of lipopolysaccharide binding protein, a marker of microbial translocation, associated with higher diversity and relative abundance of Clostridiales and an unknown genus belonging to the Cyanobacteria phylum compared to patients without ascites. The composition of the fecal microbiome was also more altered in patients with than without ascites, confirming previous studies on fecal microbiome. We propose that alteration of the serum and fecal microbiome composition be considered indicators of cirrhosis progression.


Asunto(s)
Líquido Ascítico/microbiología , Bacterias/clasificación , Heces/microbiología , Cirrosis Hepática/microbiología , Suero/microbiología , Proteínas de Fase Aguda , Bacterias/genética , Proteínas Portadoras/sangre , Progresión de la Enfermedad , Microbioma Gastrointestinal , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Cirrosis Hepática/sangre , Glicoproteínas de Membrana/sangre , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
13.
Sci Rep ; 5: 12693, 2015 Aug 04.
Artículo en Inglés | MEDLINE | ID: mdl-26239401

RESUMEN

The pathophysiology of irritable bowel syndrome (IBS) remains unclear. Here we investigated the microbiome of a large cohort of patients to identify specific signatures for IBS subtypes. We examined the microbiome of 113 patients with IBS and 66 healthy controls. A subset of these participants provided two samples one month apart. We analyzed a total of 273 fecal samples, generating more than 20 million 16S rRNA sequences. In patients with IBS, a significantly lower microbial diversity was associated with a lower relative abundance of butyrate-producing bacteria (P = 0.002; q < 0.06), in particular in patients with IBS-D and IBS-M. IBS patients who did not receive any treatment harboured a lower abundance of Methanobacteria compared to healthy controls (P = 0.005; q = 0.05). Furthermore, significant correlations were observed between several bacterial taxa and sensation of flatulence and abdominal pain (P < 0.05). Altogether, our findings showed that IBS-M and IBS-D patients are characterized by a reduction of butyrate producing bacteria, known to improve intestinal barrier function, and a reduction of methane producing microorganisms a major mechanism of hydrogen disposal in the human colon, which could explain excess of abdominal gas in IBS.


Asunto(s)
Dolor Abdominal/microbiología , Butiratos/metabolismo , Euryarchaeota/metabolismo , Flatulencia/microbiología , Síndrome del Colon Irritable/microbiología , Metano/biosíntesis , Dolor Abdominal/patología , Dolor Abdominal/fisiopatología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Bacteroides/clasificación , Bacteroides/genética , Bacteroides/metabolismo , Estudios de Casos y Controles , Euryarchaeota/clasificación , Euryarchaeota/genética , Heces/microbiología , Femenino , Firmicutes/clasificación , Firmicutes/genética , Firmicutes/metabolismo , Flatulencia/patología , Flatulencia/fisiopatología , Variación Genética , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Síndrome del Colon Irritable/patología , Síndrome del Colon Irritable/fisiopatología , Masculino , Microbiota/genética , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , ARN Ribosómico 16S/genética
14.
BMC Microbiol ; 14: 112, 2014 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-24884524

RESUMEN

BACKGROUND: The microbial community analysis of stools requires optimised and standardised protocols for their collection, homogenisation, microbial disruption and nucleic acid extraction. Here we examined whether different layers of the stool are equally representative of the microbiome. We also studied the effect of stool water content, which typically increases in diarrhoeic samples, and of a microbial disruption method on DNA integrity and, therefore, on providing an unbiased microbial composition analysis. RESULTS: We collected faecal samples from healthy subjects and performed microbial composition analysis by pyrosequencing the V4 region of the 16S rRNA gene. To examine the effect of stool structure, we compared the inner and outer layers of the samples (N = 8). Both layers presented minor differences in microbial composition and abundance at the species level. These differences did not significantly bias the microbial community specific to an individual. To evaluate the effect of stool water content and bead-beating, we used various volumes of a water-based salt solution and beads of distinct weights before nucleic acid extraction (N = 4). The different proportions of water did not affect the UniFrac-based clustering of samples from the same subject However, the use or omission of a bead-beating step produced different proportions of Gram-positive and Gram-negative bacteria and significant changes in the UniFrac-based clustering of the samples. CONCLUSION: The degree of hydration and homogenisation of faecal samples do not significantly alter their microbial community composition. However, the use of bead-beating is critical for the proper detection of Gram-positive bacteria such as Blautia and Bifidobacterium.


Asunto(s)
Bacterias/clasificación , Bacterias/aislamiento & purificación , Biota , Heces/microbiología , Manejo de Especímenes/métodos , Manejo de Especímenes/normas , Bacterias/genética , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Voluntarios Sanos , Humanos , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
15.
PLoS One ; 9(4): e95476, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24748167

RESUMEN

From birth onwards, the human gut microbiota rapidly increases in diversity and reaches an adult-like stage at three years of age. After this age, the composition may fluctuate in response to external factors such as antibiotics. Previous studies have shown that resilience is not complete months after cessation of the antibiotic intake. However, little is known about the short-term effects of antibiotic intake on the gut microbial community. Here we examined the load and composition of the fecal microbiota immediately after treatment in 21 patients, who received broad-spectrum antibiotics such as fluoroquinolones and ß-lactams. A fecal sample was collected from all participants before treatment and one week after for microbial load and community composition analyses by quantitative PCR and pyrosequencing of the 16S rRNA gene, respectively. Fluoroquinolones and ß-lactams significantly decreased microbial diversity by 25% and reduced the core phylogenetic microbiota from 29 to 12 taxa. However, at the phylum level, these antibiotics increased the Bacteroidetes/Firmicutes ratio (p = 0.0007, FDR = 0.002). At the species level, our findings unexpectedly revealed that both antibiotic types increased the proportion of several unknown taxa belonging to the Bacteroides genus, a Gram-negative group of bacteria (p = 0.0003, FDR<0.016). Furthermore, the average microbial load was affected by the treatment. Indeed, the ß-lactams increased it significantly by two-fold (p = 0.04). The maintenance of or possible increase detected in microbial load and the selection of Gram-negative over Gram-positive bacteria breaks the idea generally held about the effect of broad-spectrum antibiotics on gut microbiota.


Asunto(s)
Antibacterianos/farmacología , Tracto Gastrointestinal/efectos de los fármacos , Tracto Gastrointestinal/microbiología , Microbiota/efectos de los fármacos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Bacterias/clasificación , Bacterias/genética , Biodiversidad , Biomasa , Heces/microbiología , Femenino , Humanos , Masculino , Metagenoma , Persona de Mediana Edad , Filogenia , ARN Ribosómico 16S , Adulto Joven
16.
J Innate Immun ; 6(3): 263-71, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24296725

RESUMEN

BACKGROUND: The cross talk between the gut microbiota and the immune system, which is essential to maintain homeostasis, takes place at the intestinal lymphoid tissue such as the mesenteric lymph nodes (MLNs). Here, we investigated the presence of bacterial DNA in MLNs of control and cirrhotic rats and its relationship with inflammatory responses. METHODS: The MLN microbiome of cirrhotic rats with ascites, which was induced by carbon tetrachloride (CCl4), was compared to that of control rats using quantitative real-time PCR and pyrosequencing of the 16S rRNA gene. Cytokines in blood samples were assessed by ELISA. RESULTS: Unexpectedly, sequence analysis revealed a high microbial diversity in the MLNs of both control and cirrhotic rats with Proteobacteria as one of the most dominant phylum. CCl4-induced liver injury was not associated with a change in bacterial load, but it was linked to a decrease in microbial diversity (p < 0.05) and alterations in the microbial community in MLNs. A high proportion of Bifidobacterium animalis was also positively correlated with elevated interleukin-10 expression (p = 0.002, false discovery rate = 0.03, r = 0.94). CONCLUSIONS: For the first time, the high microbial diversity observed in MLNs of both controls and CCl4-induced cirrhotic rats provides evidence that bacterial translocation is more than a mere dichotomic phenomenon.


Asunto(s)
Bifidobacterium/inmunología , Disbiosis/inmunología , Disbiosis/microbiología , Fibrosis/inmunología , Fibrosis/microbiología , Interleucina-10/metabolismo , Ganglios Linfáticos/inmunología , Mesenterio/patología , Proteobacteria/inmunología , Animales , Bifidobacterium/genética , Biodiversidad , Tetracloruro de Carbono/administración & dosificación , Células Cultivadas , ADN Bacteriano/análisis , Modelos Animales de Enfermedad , Disbiosis/inducido químicamente , Fibrosis/inducido químicamente , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Interleucina-10/genética , Ganglios Linfáticos/microbiología , Ganglios Linfáticos/patología , Masculino , Microbiota/genética , Proteobacteria/genética , ARN Ribosómico 16S/genética , Ratas , Ratas Sprague-Dawley , Regulación hacia Arriba
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...