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1.
Microbiol Spectr ; 9(3): e0080221, 2021 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-34787462

RESUMO

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related deaths worldwide. Although dysbiosis of the lung and gut microbiota have been associated with NSCLC, their relative contributions are unclear; in addition, their roles in distant metastasis (DM) are still illusive. We recruited in total 121 participants, including 87 newly diagnosed treatment-naive NSCLC patients of various stages and 34 healthy volunteers, and surveyed their fecal and sputum microbiota. We compared the microbial profiles between groups, identified microbial biomarkers, and generated machine learning models for distinguishing healthy individuals from patients with NSCLC and patients of various stages. We found significant perturbations of gut and sputum microbiota in patients with NSCLC and DM. A machine learning model combining both microbiota (combined model) performed better than an individual data set in patient stratification, with the highest area under the curve (AUC) value of 0.896. Sputum and gut microbiota both contributed to the combined model; in most cases, sputum-only models performed similar to the combined models. Several microbial biomarkers were shared by both microbiotas, indicating their similar roles at distinct body sites. Microbial biomarkers of distinct disease stages were mostly shared, suggesting biomarkers for DM could be acquired early. Furthermore, Pseudomonas aeruginosa, a species previously associated with wound infections, was significantly more abundant in brain metastasis, indicating that distinct types of DMs could have different microbes. Our results indicate that alterations of the sputum microbiota have stronger relationships with NSCLC and DM than the gut and strongly support the feasibility of metagenome-based noninvasive disease diagnosis and risk evaluation. (This study has been registered at ClinicalTrials.gov under registration no. NCT03454685). IMPORTANCE Our survey on gut and sputum microbiota revealed that both were significantly disturbed in non-small cell lung cancer (NSCLC) and associated with distant metastasis (DM) while only the sputum microbiota was associated with non-DM NSCLC. The lung microbiota could therefore have a stronger association with (and thus may contribute more to) disease development than the gut microbiota. Mathematic models using both microbiotas performed better in patient stratification than using individual microbiota. Sputum models, however, performed similar to the combined models, suggesting a convenient, noninvasive diagnostic for NSCLC. Microbial biomarkers of distinct disease stages were mostly shared, suggesting that the same set of microbes were underlying disease progression, and the signals for distant metastasis could be acquired at early stages of the disease. Our results strongly support the feasibility of noninvasive diagnosis of NSCLC, including distant metastasis, are of clinical importance, and should warrant further research on the underlying molecular mechanisms.


Assuntos
Bactérias/classificação , Carcinoma Pulmonar de Células não Pequenas/patologia , Disbiose/microbiologia , Microbioma Gastrointestinal/fisiologia , Neoplasias Pulmonares/patologia , Pulmão/microbiologia , Actinobacteria/isolamento & purificação , Bactérias/isolamento & purificação , Bacteroidetes/isolamento & purificação , Biomarcadores , Fezes/microbiologia , Feminino , Firmicutes/isolamento & purificação , Fusobactérias/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica/diagnóstico , Metástase Neoplásica/patologia , Proteobactérias/isolamento & purificação , Escarro/microbiologia
2.
Microbiologyopen ; 10(4): e1215, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34459554

RESUMO

As apex predators, pinnipeds are considered to be useful bioindicators of marine and coastal environments. Endemic to a small archipelago in the South Pacific, the Juan Fernandez fur seal (JFFS) is one of the less-studied members of the pinniped family Otariidae. This study aimed to characterize the fecal microbiome of the JFFS for the first time, to establish a baseline for future studies of host-microbial-environment interactions and monitoring programs. During two consecutive reproductive seasons, 57 fecal samples were collected from seven different JFFS colonies within the Juan Fernandez Archipelago, Chile. Bacterial composition and abundance were characterized by sequencing the V4 region of the 16S rRNA gene. The overall microbiome composition was dominated by five phyla: Firmicutes (40% ±24), Fusobacteria (30% ±17), Bacteroidetes (22% ±10), Proteobacteria (6% ±4), and Actinobacteria (2% ±3). Alpha diversity was higher in Tierras Blancas. However, location was not found to be a dominant driver of microbial composition. Interestingly, the strongest signal in the data was a negative association between the genera Peptoclostridium and Fusobacterium, which explained 29.7% of the total microbial composition variability between samples. The genus Peptoclostridium has not been reported in other pinniped studies, and its role here is unclear, with interpretation challenging due to a lack of information regarding microbiome functionality in marine mammals. As a first insight into the JFFS fecal microbiome, these results contribute towards our understanding of the natural microbial diversity and composition in free-ranging pinnipeds.


Assuntos
Bactérias/classificação , Fezes/microbiologia , Otárias/microbiologia , Microbioma Gastrointestinal/genética , Microbiota/genética , Actinobacteria/classificação , Actinobacteria/genética , Actinobacteria/isolamento & purificação , Animais , Bactérias/genética , Bactérias/isolamento & purificação , Bacteroidetes/classificação , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Biodiversidade , Chile , DNA Bacteriano/genética , Firmicutes/classificação , Firmicutes/genética , Firmicutes/isolamento & purificação , Fusobactérias/classificação , Fusobactérias/genética , Fusobactérias/isolamento & purificação , Proteobactérias/classificação , Proteobactérias/genética , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
3.
Biomolecules ; 11(8)2021 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-34439821

RESUMO

Irritable bowel syndrome is not a life-threatening disease, yet it significantly affects the quality of life and contributes to economic loss. It is estimated that even up to 45% of the world's population can suffer from the disease. The first attempts to diagnose irritable bowel syndrome were made at the end of the 19th century; however, establishing appropriate diagnostic criteria and treatment methods is still ongoing. To date, little is known about the etiology of irritable bowel syndrome; however, growing attention is drawn to the intestinal microbiota as a factor in the disease development. For this reason, researchers have conducted many studies on therapies that modulate the microbiota, among which probiotics, prebiotics, and synbiotics are widely studied. To date, most studies have examined probiotics; however, there are also several studies demonstrating the efficacy of prebiotics and synbiotics. The aim of this review was to summarize findings on the usefulness of probiotics, prebiotics, and synbiotics in the treatment of irritable bowel syndrome.


Assuntos
Disbiose/dietoterapia , Microbioma Gastrointestinal/efeitos dos fármacos , Síndrome do Intestino Irritável/dietoterapia , Prebióticos/administração & dosagem , Probióticos/administração & dosagem , Simbióticos/administração & dosagem , Actinobacteria/genética , Actinobacteria/crescimento & desenvolvimento , Actinobacteria/isolamento & purificação , Antibacterianos/efeitos adversos , Bacteroidetes/genética , Bacteroidetes/crescimento & desenvolvimento , Bacteroidetes/isolamento & purificação , Ensaios Clínicos como Assunto , Disbiose/etiologia , Disbiose/microbiologia , Disbiose/patologia , Firmicutes/genética , Firmicutes/crescimento & desenvolvimento , Firmicutes/isolamento & purificação , Fusobactérias/genética , Fusobactérias/crescimento & desenvolvimento , Fusobactérias/isolamento & purificação , Microbioma Gastrointestinal/genética , Humanos , Síndrome do Intestino Irritável/microbiologia , Síndrome do Intestino Irritável/patologia , Proteobactérias/genética , Proteobactérias/crescimento & desenvolvimento , Proteobactérias/isolamento & purificação , Qualidade de Vida
4.
Genome Biol ; 22(1): 192, 2021 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-34183041

RESUMO

A critical challenge in microbiome data analysis is the existence of many non-biological zeros, which distort taxon abundance distributions, complicate data analysis, and jeopardize the reliability of scientific discoveries. To address this issue, we propose the first imputation method for microbiome data-mbImpute-to identify and recover likely non-biological zeros by borrowing information jointly from similar samples, similar taxa, and optional metadata including sample covariates and taxon phylogeny. We demonstrate that mbImpute improves the power of identifying disease-related taxa from microbiome data of type 2 diabetes and colorectal cancer, and mbImpute preserves non-zero distributions of taxa abundances.


Assuntos
Neoplasias Colorretais/microbiologia , DNA Bacteriano/genética , Diabetes Mellitus Tipo 2/microbiologia , Metagenoma , Microbiota/genética , Software , Actinobacteria/classificação , Actinobacteria/genética , Actinobacteria/isolamento & purificação , Bacteroidetes/classificação , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Estudos de Casos e Controles , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/patologia , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/patologia , Firmicutes/classificação , Firmicutes/genética , Firmicutes/isolamento & purificação , Fusobactérias/classificação , Fusobactérias/genética , Fusobactérias/isolamento & purificação , Humanos , Filogenia , Reação em Cadeia da Polimerase/métodos , Proteobactérias/classificação , Proteobactérias/genética , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética , Sequenciamento Completo do Genoma
5.
PLoS One ; 16(4): e0250344, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33872339

RESUMO

Canine acute hemorrhagic diarrhea syndrome (AHDS) has been associated in some studies with Clostridioides perfringens overgrowth and toxin-mediated necrosis of the intestinal mucosa. We aimed to determine the effect of a single fecal microbiota transplantation (FMT) on clinical scores and fecal microbiomes of 1 and 7 dogs with AHDS from New Zealand and South Africa. We hypothesized that FMT would improve AHDS clinical scores and increase microbiota alpha-diversity and short-chain fatty acid (SCFA)-producing microbial communities' abundances in dogs with AHDS after FMT. We sequenced the V3-V4 region of the 16S-rRNA gene in the feces of AHDS FMT-recipients and sham-treated control dogs, and their healthy donors at admission, discharge, and 30 days post-discharge. There were no significant differences in median AHDS clinical scores between FMT-recipients and sham-treated controls at admission or discharge (P = 0.22, P = 0.41). At admission, the Shannon diversity index (SDI) was lower in AHDS dogs than healthy donors (P = 0.002). The SDI did not change from admission to 30 days in sham-treated dogs yet increased in FMT-recipients from admission to discharge (P = 0.04) to levels not different than donors (P = 0.33) but significantly higher than sham-treated controls (P = 0.002). At 30 days, the SDI did not differ between FMT recipients, sham-treated controls, and donors (P = 0.88). Principal coordinate analysis of the Bray-Curtis index separated post-FMT and donor dogs from pre-FMT and sham-treated dogs (P = 0.009) because of increased SCFA-producing genera's abundances after FMT. A single co-abundance subnetwork contained many of the same OTUs found to be differentially abundant in FMT-recipients, and the abundance of this module was increased in FMT-recipients at discharge and 30 days, compared to sham-treated controls. We conclude in this small pilot study FMT did not have any clinical benefit. A single FMT procedure has the potential to increase bacterial communities of SCFA-producing genera important for intestinal health up to 30 days post-FMT.


Assuntos
Clostridioides/patogenicidade , Infecções por Clostridium/terapia , Diarreia/terapia , Transplante de Microbiota Fecal/veterinária , Fezes/microbiologia , Hemorragia Gastrointestinal/terapia , Microbioma Gastrointestinal/fisiologia , Actinobacteria/genética , Actinobacteria/crescimento & desenvolvimento , Actinobacteria/isolamento & purificação , Animais , Bacteroidetes/genética , Bacteroidetes/crescimento & desenvolvimento , Bacteroidetes/isolamento & purificação , Clostridioides/genética , Clostridioides/crescimento & desenvolvimento , Infecções por Clostridium/microbiologia , Infecções por Clostridium/patologia , Infecções por Clostridium/veterinária , Diarreia/microbiologia , Diarreia/patologia , Diarreia/veterinária , Cães , Ácidos Graxos Voláteis/biossíntese , Feminino , Firmicutes/genética , Firmicutes/crescimento & desenvolvimento , Firmicutes/isolamento & purificação , Fusobactérias/genética , Fusobactérias/crescimento & desenvolvimento , Fusobactérias/isolamento & purificação , Hemorragia Gastrointestinal/microbiologia , Hemorragia Gastrointestinal/patologia , Hemorragia Gastrointestinal/veterinária , Mucosa Intestinal/microbiologia , Masculino , Nova Zelândia , Projetos Piloto , Estudos Prospectivos , Proteobactérias/genética , Proteobactérias/crescimento & desenvolvimento , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética , África do Sul
6.
Crit Rev Microbiol ; 47(4): 517-542, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33823747

RESUMO

Sneathia is an emerging pathogen implicated in adverse reproductive and perinatal outcomes. Although scarce, recent data suggest that vaginally residing Sneathia becomes pathogenic following its ascension into the upper urogenital tract, amniotic fluid, placenta, and foetal membranes. The role of Sneathia in women's health and disease is generally underappreciated because the cultivation of these bacteria is limited by their complex nutritional requirements, slow growth patterns, and anaerobic nature. For this reason, molecular methods are typically required for the detection and differential diagnosis of Sneathia infections. Here, we review the laboratory methods used for the diagnosis of Sneathia infections, the molecular mechanisms underlying its virulence, and its sensitivity to antibiotics. We further review the evidence of Sneathia's contributions to the pathogenesis of bacterial vaginosis, chorioamnionitis, preterm prelabour rupture of membranes, spontaneous preterm labour, stillbirth, maternal and neonatal sepsis, HIV infection, and cervical cancer. Collectively, growing evidence indicates that Sneathia represents an important yet underappreciated pathogen affecting the development and progression of several adverse clinical conditions diagnosed in pregnant women and their neonates, as well as in non-pregnant women.


Assuntos
Fusobactérias/fisiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Complicações Infecciosas na Gravidez/microbiologia , Vaginose Bacteriana/microbiologia , Animais , Feminino , Fusobactérias/genética , Fusobactérias/isolamento & purificação , Humanos , Gravidez
7.
Cell Rep Med ; 2(3): 100206, 2021 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-33763652

RESUMO

Extremely low birth weight (ELBW) infants often develop an altered gut microbiota composition, which is related to clinical complications, such as necrotizing enterocolitis and sepsis. Probiotic supplementation may reduce these complications, and modulation of the gut microbiome is a potential mechanism underlying the probiotic effectiveness. In a randomized, double-blind, placebo-controlled trial, we assessed the effect of Lactobacillus reuteri supplementation, from birth to post-menstrual week (PMW)36, on infant gut microbiota. We performed 16S amplicon sequencing in 558 stool samples from 132 ELBW preterm infants at 1 week, 2 weeks, 3 weeks, 4 weeks, PMW36, and 2 years. Probiotic supplementation results in increased bacterial diversity and increased L. reuteri abundance during the 1st month. At 1 week, probiotic supplementation also results in a lower abundance of Enterobacteriaceae and Staphylococcaceae. No effects were found at 2 years. In conclusion, probiotics may exert benefits by modulating the gut microbiota composition during the 1st month in ELBW infants.


Assuntos
Suplementos Nutricionais , Microbioma Gastrointestinal/genética , Recém-Nascido de Peso Extremamente Baixo ao Nascer/crescimento & desenvolvimento , Lactente Extremamente Prematuro/crescimento & desenvolvimento , Limosilactobacillus reuteri/fisiologia , Probióticos/administração & dosagem , Actinobacteria/classificação , Actinobacteria/genética , Actinobacteria/isolamento & purificação , Bacteroidetes/classificação , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Biodiversidade , Fezes/microbiologia , Feminino , Firmicutes/classificação , Firmicutes/genética , Firmicutes/isolamento & purificação , Fusobactérias/classificação , Fusobactérias/genética , Fusobactérias/isolamento & purificação , Humanos , Lactente , Masculino , Proteobactérias/classificação , Proteobactérias/genética , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética , Verrucomicrobia/classificação , Verrucomicrobia/genética , Verrucomicrobia/isolamento & purificação
8.
Medicine (Baltimore) ; 99(41): e22494, 2020 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-33031285

RESUMO

INTRODUCTION: Sneathia sanguinegens(S sanguinegens) is a gram-negative rod-shaped bacterium mostly reported to cause a perinatal infection, and there are no reports of S sanguinegens in prosthetic joint infection (PJI). The purpose of this report is to describe a very rare case of PJI after total hip arthroplasty (THA) caused by S sanguinegens. PATIENT CONCERNS: A 79-year-old woman presented with right coxalgia, inability to walk, and a fever of 39°C. She had undergone THA 28 years earlier for osteoarthritis of the hip. DIAGNOSES: The diagnosis was acute late-onset PJI, because blood tests revealed marked inflammatory reaction and computed tomography showed an abscess at the right hip joint; synovial fluid analysis resulted in detection of a gram-negative bacillus. INTERVENTION: Surgical debridement with retention of the implant and antibiotic therapy was performed. OUTCOMES: One month after surgery, polymerase chain reaction (PCR) assay showed that the pathogen was 99.9% likely to be S sanguinegens. There has been no recurrence of infection or loosening of the implant in the 2 years since her surgery. LESSONS: PCR should facilitate detection of previously unknown pathogens and potentially novel bacterial species.


Assuntos
Fusobactérias , Infecções por Bactérias Gram-Negativas/complicações , Infecções Relacionadas à Prótese/microbiologia , Idoso , Antibacterianos/uso terapêutico , Artroplastia de Quadril , Feminino , Fusobactérias/isolamento & purificação , Infecções por Bactérias Gram-Negativas/diagnóstico , Humanos , Reação em Cadeia da Polimerase , Infecções Relacionadas à Prótese/diagnóstico por imagem , Infecções Relacionadas à Prótese/cirurgia
9.
Anaerobe ; 66: 102277, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32987144

RESUMO

Sneathia amnii is an opportunistic pathogen of the female reproductive tract that has been reported to cause infections during pregnancy and in the post-partum period. Infections outside the reproductive tract have rarely been described. We report the case of a spondylitis due to S. amnii in a 72-year old woman, successfully treated after seven weeks of antimicrobial therapy. Growth of this pathogen guided our diagnosis towards a gynecological pathology; we discovered an endometrium adenocarcinoma. This case emphasizes the need for adequate incubation of discal biopsies, using aerobic and anaerobic enrichment broth with prolonged incubation.


Assuntos
Antibacterianos/uso terapêutico , Fusobactérias/classificação , Espondilite/diagnóstico , Espondilite/microbiologia , Adenocarcinoma , Idoso , DNA Bacteriano , Neoplasias do Endométrio , Feminino , Fusobactérias/efeitos dos fármacos , Fusobactérias/isolamento & purificação , Humanos , RNA Ribossômico 16S , Espondilite/tratamento farmacológico , Resultado do Tratamento
10.
AIDS ; 34(10): 1451-1460, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32675558

RESUMO

OBJECTIVES: The aim of this study was to examine the relationship between gut microbial communities in HIV-infected individuals on suppressive antiretroviral therapy (cART), and the peripheral HIV-Gag-specific CD8 T-cell responses before and after ex-vivo immune checkpoint blockade (ICB). DESIGN: Thirty-four HIV-seropositive, 10 HIV-seronegative and 12 HIV-seropositive receiving faecal microbiota transplant (FMT) participants were included. Gut microbial communities, peripheral and gut associated negative checkpoint receptors (NCRs) and peripheral effector functions were assessed. METHODS: Bacterial 16s rRNA sequencing for gut microbiome study and flow-based assays for peripheral and gut NCR and their cognate ligand expression, including peripheral HIV-Gag-specific CD8 T-cell responses before and after ex-vivo anti-PD-L1 and anti-TIGIT ICB were performed. RESULTS: Fusobacteria abundance was significantly higher in HIV-infected donors compared to uninfected controls. In HIV-infected participants receiving Fusobacteria-free FMT, Fusobacteria persisted up to 24 weeks in stool post FMT. PD-1 TIGIT and their ligands were expanded in mucosal vs. peripheral T cells and dendritic cells, respectively. PD-L1 and TIGIT blockade significantly increased the magnitude of peripheral anti-HIV-Gag-specific CD8 T-cell responses. Higher gut Fusobacteria abundance was associated with lower magnitude of peripheral IFN-γ+ HIV-Gag-specific CD8 T-cell responses following ICB. CONCLUSION: The gut colonization of Fusobacteria in HIV infection is persistent and may influence anti-HIV T-cell immunity to PD-1 or TIGIT blockade. Strategies modulating Fusobacteria colonization may elicit a favourable mucosal immune landscape to enhance the efficacy of ICB for HIV cure.


Assuntos
Linfócitos T CD8-Positivos/efeitos dos fármacos , Microbioma Gastrointestinal , Infecções por HIV/microbiologia , Inibidores de Checkpoint Imunológico/uso terapêutico , Adulto , Idoso , Fármacos Anti-HIV/uso terapêutico , Transplante de Microbiota Fecal , Feminino , Fusobactérias/isolamento & purificação , Infecções por HIV/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-Idade , RNA Ribossômico 16S/genética , Minorias Sexuais e de Gênero
11.
Int J Syst Evol Microbiol ; 70(5): 3037-3048, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32223835

RESUMO

Two independent strains of a Leptotrichia species (ES3154-GLUT and ES2714_GLU) were isolated from the oral cavity of northern elephant seals (Mirounga angustirostris) that were admitted to The Marine Mammal Centre facilities in California, USA. The strains were isolated from oral swabs by cultivation in PPLO broth supplemented with serum, penicillin and colistin in anaerobic conditions. The strains were Gram-negative, pleomorphic, indole-, oxidase- and catalase-negative, non-spore-forming, non-motile rods/coccobacilli in short chains. The 16S rRNA gene sequence of these strains shared 94.42 % nucleotide similarity with Oceanivirga salmonicida AVG 2115T but demonstrated ≤86.00-92.50 % nucleotide similarity to the 16S rRNA genes of other species of the family Leptotrichiaceae. The genome was sequenced for strain ES3154-GLUT. Average nucleotide identity values between strain ES3154-GLUT and 15 type strain genomes from the family Leptotrichiaceae ranged from 66.74 % vs. Sebaldella termitidis to 73.35 % vs. O. salmonicida. The whole genome phylogeny revealed that the novel species was most closely related to O. salmonicida AVG 2115T. This relationship was also confirmed by nucleotide similarity and multilocus phylogenetic analyses employing various housekeeping genes (partial 23S rRNA, rpoB, rpoC, rpoD, polC, adh, gyrA and gyrB genes). Chemotaxonomic and phenotypical features of strain ES3154-GLUT were in congruence with closely related members of the family Leptotrichiaceae, represented by similar enzyme profiles and fatty acid patterns. MALDI-TOF MS analysis was capable to clearly discriminate strain ES3154-GLUT from all currently described taxa of the family Leptotrichiaceae. Based on these data, we propose a novel species of the genus Oceanivirga, for which the name Oceanivirga miroungae sp. nov. is proposed with the type strain ES3154-GLUT (=DSM 109740T=CCUG 73653T=ATCC TSD-189T=NCTC 14411T) and one representative strain ES2714_GLU. The G+C content is 26.82 %, genome size is 1 356 983 bp.


Assuntos
Fusobactérias/classificação , Boca/microbiologia , Filogenia , Focas Verdadeiras/microbiologia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , California , DNA Bacteriano/genética , Ácidos Graxos/química , Fusobactérias/isolamento & purificação , Genes Bacterianos , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , RNA Ribossômico 23S , Análise de Sequência de DNA
12.
J Crohns Colitis ; 14(11): 1535-1546, 2020 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-32333762

RESUMO

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.


Assuntos
Doença de Crohn , Disbiose , Fusobactérias/isolamento & purificação , Microbioma Gastrointestinal/fisiologia , Mucosa Intestinal , Complicações Pós-Operatórias , Adulto , Bélgica/epidemiologia , Biópsia/métodos , Ceco/cirurgia , Doença de Crohn/epidemiologia , Doença de Crohn/microbiologia , Doença de Crohn/patologia , Doença de Crohn/cirurgia , Procedimentos Cirúrgicos do Sistema Digestório/efeitos adversos , Procedimentos Cirúrgicos do Sistema Digestório/métodos , Disbiose/diagnóstico , Disbiose/etiologia , Disbiose/microbiologia , Fezes/química , Fezes/microbiologia , Feminino , Humanos , Íleo/cirurgia , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Masculino , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/microbiologia , Recidiva , Resultado do Tratamento
13.
Nat Microbiol ; 5(4): 642-650, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32042128

RESUMO

Although Clostridium difficile is widely considered an antibiotic- and hospital-associated pathogen, recent evidence indicates that this is an insufficient depiction of the risks and reservoirs. A common thread that links all major risk factors of infection is their association with gastrointestinal disturbances, but this relationship to C. difficile colonization has never been tested directly. Here, we show that disturbances caused by diarrhoeal events trigger susceptibility to C. difficile colonization. Using survey data of the human gut microbiome, we detected C. difficile colonization and blooms in people recovering from food poisoning and Vibrio cholerae infections. Carriers remained colonized for year-long time scales and experienced highly variable patterns of C. difficile abundance, where increased shedding over short periods of 1-2 d interrupted week-long periods in which C. difficile was undetectable. Given that short shedding events were often linked to gastrointestinal disturbances, our results help explain why C. difficile is frequently detected as a co-infecting pathogen in patients with diarrhoea. To directly test the impact of diarrhoea on susceptibility to colonization, we developed a mouse model of variable disturbance intensity, which allowed us to monitor colonization in the absence of disease. As mice exposed to avirulent C. difficile spores ingested increasing quantities of laxatives, more individuals experienced C. difficile blooms. Our results indicate that the likelihood of colonization is highest in the days immediately following acute disturbances, suggesting that this could be an important window during which transmission could be interrupted and the incidence of infection lowered.


Assuntos
Clostridioides difficile/efeitos dos fármacos , Clostridioides difficile/patogenicidade , Infecções por Clostridium/microbiologia , Diarreia/microbiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Laxantes/efeitos adversos , Polietilenoglicóis/efeitos adversos , Actinobacteria/genética , Actinobacteria/crescimento & desenvolvimento , Actinobacteria/isolamento & purificação , Animais , Bacteroidetes/genética , Bacteroidetes/crescimento & desenvolvimento , Bacteroidetes/isolamento & purificação , Clostridioides difficile/crescimento & desenvolvimento , Infecções por Clostridium/complicações , Contagem de Colônia Microbiana , Diarreia/induzido quimicamente , Diarreia/complicações , Modelos Animais de Doenças , Fezes/microbiologia , Firmicutes/genética , Firmicutes/crescimento & desenvolvimento , Firmicutes/isolamento & purificação , Fusobactérias/genética , Fusobactérias/crescimento & desenvolvimento , Fusobactérias/isolamento & purificação , Humanos , Masculino , Camundongos , Proteobactérias/genética , Proteobactérias/crescimento & desenvolvimento , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética
14.
Microbiol Immunol ; 64(2): 99-112, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31793046

RESUMO

Periodontitis is a major cause of tooth loss in adults that initially results from dental plaque. Subgingival plaque pathogenesis is affected by both community composition and plaque structures, although limited data are available concerning the latter. To bridge this knowledge gap, subgingival plaques were obtained using filter paper (the fourth layer) and curette (the first-third layers) sequentially and the phylogenetic differences between the first-third layers and the fourth layer were characterized by sequencing the V3-V4 regions of 16S rRNA. A total of 11 phyla, 148 genera, and 308 species were obtained by bioinformatic analysis, and no significant differences between the operational taxonomic unit numbers were observed for these groups. In both groups, the most abundant species were Porphyromonas gingivalis and Fusobacterium nucleatum. Actinomyces naeslundii, Streptococcus intermedius, and Prevotella intermedia possessed relatively high proportions in the first-third layers; while in the fourth layer, both traditional pathogens (Treponema denticola and Campylobacter rectus) and novel pathobionts (Eubacterium saphenum, Filifactor alocis, Treponema sp. HOT238) were prominent. Network analysis showed that either of them exhibited a scale-free property and was constructed by two negatively correlated components (the pathogen component and the nonpathogen component), while the synergy in the nonpathogen component was lower in the first-third layers than that in the fourth layer. After merging these two parts into a whole plaque group, the negative/positive correlation ratio increased. With potential connections, the first-third layers and the fourth layer showed characteristic key nodes in bacterial networks.


Assuntos
Bactérias/isolamento & purificação , Placa Dentária/microbiologia , Microbiota , Periodontite/microbiologia , Actinobacteria/classificação , Actinobacteria/genética , Actinobacteria/isolamento & purificação , Actinomyces/isolamento & purificação , Adulto , Bactérias/classificação , Bactérias/genética , Classificação , Feminino , Fusobactérias/classificação , Fusobactérias/genética , Fusobactérias/isolamento & purificação , Fusobacterium/isolamento & purificação , Fusobacterium nucleatum/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Masculino , Metagenômica , Microbiota/genética , Filogenia , Porphyromonas gingivalis/isolamento & purificação , Prevotella intermedia/isolamento & purificação , RNA Ribossômico 16S/genética , Spirochaetales/classificação , Spirochaetales/genética , Spirochaetales/isolamento & purificação , Streptococcus intermedius/isolamento & purificação , Treponema/isolamento & purificação , Adulto Jovem
15.
Microb Pathog ; 139: 103851, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31715320

RESUMO

BACKGROUND: The presence of the human lung microbiota has been demonstrated in patients with different lung diseases, mainly in sputum samples. However, for study of the alveolar microbiota, a bronchoalveolar lavage (BAL) sample represents the lower respiratory tract (LRT) environment. It is currently unknown whether there is a specific alveolar microbiota profile in human lung diseases, such as pulmonary tuberculosis (TB) and interstitial pneumonia (IP). METHODS: BAL samples from six active TB patients, six IP patients and ten healthy volunteers were used for DNA extraction followed by amplification of the complete bacterial 16S ribosomal RNA gene (16S rDNA). The 16S rDNA was sequenced with a MiSeq Desktop Sequencer, and the data were analysed by QIIME software for taxonomic assignment. RESULTS: The alveolar microbiota in TB and IP patients and healthy volunteers was characterized by six dominant phyla, Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, Fusobacteria and Cyanobacteria. A significant reduction in the abundance of Firmicutes was observed in IP patients. In TB and IP patients, the diversity of the alveolar microbiota was diminished, characterized by a significant reduction in the abundance of the Streptococcus genus and associated with increased Mycobacterium abundance in TB patients and diminished Acinetobacter abundance in IP patients with respect to their abundances in healthy volunteers. However, an important difference was observed between TB and IP patients: the Fusobacterium abundance was significantly reduced in TB patients. Exclusive genera that were less abundant in patients than in healthy volunteers were characterized for each study group. CONCLUSIONS: This study shows that the alveolar microbiota profile in BAL samples from TB and IP patients, representing infectious and non-infectious lung diseases, respectively, is characterized by decreased diversity.


Assuntos
Doenças Pulmonares Intersticiais/microbiologia , Microbiota , Tuberculose Pulmonar/microbiologia , Actinobacteria/isolamento & purificação , Actinobacteria/metabolismo , Adulto , Idoso , Bacteroidetes/isolamento & purificação , Bacteroidetes/metabolismo , Lavagem Broncoalveolar , Cianobactérias/isolamento & purificação , Cianobactérias/metabolismo , Feminino , Firmicutes/isolamento & purificação , Firmicutes/metabolismo , Fusobactérias/isolamento & purificação , Fusobactérias/metabolismo , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Proteobactérias/isolamento & purificação , Proteobactérias/metabolismo , RNA Bacteriano/genética , RNA Bacteriano/isolamento & purificação , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/isolamento & purificação , Sistema Respiratório/microbiologia , Escarro/microbiologia , Adulto Jovem
16.
Appl Microbiol Biotechnol ; 103(21-22): 8977-8985, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31630239

RESUMO

The Australian saltwater crocodile (Crocodylus porosus) industry began commercially in the 1980s, producing skins for export and crocodile meat as a by-product. Industry research has thus far focused on strategies to improve production efficiency. In the current study, we utilised 16S rRNA sequencing to characterise the intestinal microbiome of Australian saltwater crocodiles. Samples were collected from 13 commercially farmed crocodiles from six sample sites along the length of the intestinal tract. The results indicate a similar microbiome composition to that found in the freshwater alligator, with the dominate phyla represented by Firmicutes, primarily Clostridia, and Fusobacteria, which appears to be distinct from mammalian, fish, and other reptile phyla which are generally dominated by Firmicutes and Bacteroidetes. The high abundance of 'pathogenic' bacteria, with no apparent consequence to the host's health, is of great interest and warrants further additional investigation. This will enable expansion of the current understanding of host immune function and how it is modified by host and intestinal microbiome interactions.


Assuntos
Jacarés e Crocodilos/microbiologia , Clostridiales/isolamento & purificação , Fusobactérias/isolamento & purificação , Microbioma Gastrointestinal/genética , Intestinos/microbiologia , Animais , Austrália , Sequência de Bases , Clostridiales/classificação , Clostridiales/genética , Fusobactérias/classificação , Fusobactérias/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
17.
J Dairy Sci ; 102(12): 11786-11797, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31587913

RESUMO

Until 2010, our knowledge of the uterine microbiome in cows that developed uterine disease relied almost exclusively on culture-dependent studies and mostly included cows with clinical endometritis (i.e., with purulent uterine discharge). Those studies consistently found a strong positive correlation between Trueperella pyogenes and clinical endometritis, whereas other pathogens such as Escherichia coli, Fusobacterium necrophorum, Prevotella melaninogenica, and Bacteroides spp. were also commonly cocultured. In contrast, Streptococcus spp., Staphylococcus spp., and Bacillus spp. were usually isolated from healthy cows. Starting in 2010, culture-independent studies using PCR explored the microbiome of cows with metritis and clinical endometritis, and observed that E. coli was a pioneer pathogen that predisposed cows to infection with F. necrophorum, which was strongly associated with metritis, and to infection with T. pyogenes, which was strongly associated with clinical endometritis. Starting in 2011, culture-independent studies using metagenomic sequencing expanded our knowledge of the uterine microbiome. It has been shown that cows have bacteria in the uterus even before calving, they have an established uterine microbiome within 20 min of calving, and that the microbiome structure is identical between cows that develop metritis and healthy cows until 2 d postpartum, after which the bacterial structure of cows that developed metritis deviates in favor of greater relative abundance of Bacteroidetes and Fusobacteria and lesser relative abundance of Proteobacteria and Tenericutes. The shift in the uterine microbiome in cows that develop metritis is characterized by a loss of heterogeneity and a decrease in bacterial richness. At the genus level, Bacteroides, Porphyromonas, and Fusobacterium have the strongest association with metritis. At the species level, we observed that Bacteroides pyogenes, Porphyromonas levii, and Helcococcus ovis were potential emerging uterine pathogens. Finally, we have shown that the hematogenous route is a viable route of uterine infection with uterine pathogens. Herein, we propose that metritis is associated with a dysbiosis of the uterine microbiota characterized by decreased richness, and an increase in Bacteroidetes and Fusobacteria, particularly Bacteroides, Porphyromonas, and Fusobacterium.


Assuntos
Bactérias/isolamento & purificação , Doenças dos Bovinos/microbiologia , Endometrite/veterinária , Microbiota , Doenças Uterinas/veterinária , Animais , Bactérias/genética , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Bovinos , Doenças dos Bovinos/patologia , Disbiose/microbiologia , Disbiose/veterinária , Endometrite/microbiologia , Feminino , Fusobactérias/genética , Fusobactérias/isolamento & purificação , Reação em Cadeia da Polimerase , Período Pós-Parto , Doenças Uterinas/microbiologia , Doenças Uterinas/patologia , Útero/microbiologia
18.
Sci Rep ; 9(1): 9084, 2019 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-31235833

RESUMO

Kwashiorkor and marasmus are considered to be two different clinical diseases resulting from severe malnutrition, but this distinction has been questioned. In a previous study comparing children with kwashiorkor and healthy children from Niger and Senegal, we found a dramatic gut microbiota alteration with a predominant depletion of anaerobes and enrichment in Proteobacteria and Fusobacteria in kwashiorkor. However, it remained unknown whether this association was related to malnutrition or was a specific feature of kwashiorkor. In this continuation study, we added 7 new marasmus subjects and 71,162 new colonies from the same countries. Our results showed that, compared to marasmus, the kwashiorkor gut microbiota was characterized by an increased proportion of Proteobacteria (culturomics, Marasmus 5.0%, Kwashiorkor 16.7%, p < 0.0001; metagenomics, Marasmus 14.7%, Kwashiorkor 22.0%, p = 0.001), but there was a decreased proportion of Bacteroidetes in marasmus (culturomics, Marasmus 0.8%, Kwashiorkor 6.5%, p = 0.001; metagenomics, Marasmus 5.4%, Kwashiorkor 7.0%, p = 0.03). Fusobacterium was more frequently cultured from kwashiorkor. All detected potential pathogenic species were enriched in the kwashiorkor gut microbiota. These results provide a biological basis to support the usage of an antibiotic therapy more effective in suppressing the overgrowth of bacterial communities resistant to penicillin, combined with antioxidants and probiotics for nutritional recovery therapies, particularly for kwashiorkor.


Assuntos
Bacteroidetes/isolamento & purificação , Fusobactérias/isolamento & purificação , Microbioma Gastrointestinal , Kwashiorkor/microbiologia , Desnutrição Proteico-Calórica/microbiologia , Proteobactérias/isolamento & purificação , Biodiversidade , Estudos de Casos e Controles , Feminino , Humanos , Masculino
19.
Parasit Vectors ; 12(1): 252, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31113452

RESUMO

BACKGROUND: Parasitic protozoans, helminths, alter the gut microbiota in mammals, yet little is known about the influence of intestinal cestodes on gut microbiota in fish. In the present study, the composition and diversity of the hindgut microbiota were determined in the intestine of common carp (Cyprinus carpio) infected with two tapeworm species, Khawia japonensis and Atractolytocestus tenuicollis. RESULTS: The intestine contained a core microbiota composed of Proteobacteria, Fusobacteria and Tenericutes. Infection with the two cestode species had no significant effect on the microbial diversity and richness, but it altered the microbial composition at the genus level. PCoA analysis indicated that microbial communities in the infected and uninfected common carp could not be distinguished from each other. However, a Mantel test indicated that the abundance of K. japonensis was significantly correlated with the microbial composition (P = 0.015), while the abundance of A. tenuicollis was not (P = 0.954). According to Pearson's correlation analysis, the abundance of K. japonensis exhibited an extremely significant (P < 0.001) positive correlation with the following gut microbiota taxa: Epulopiscium, U114, Bacteroides, Clostridium and Peptostreptococcaceae (0.8< r < 0.9); and a significant (P < 0.05) correlation with Enterobacteriaceae, Micrococcaceae, Rummeliibacillus, Lysinibacillus boronitolerans, Veillonellaceae, Oxalobacteraceae, Aeromonadaceae (negative), Marinibacillus and Chitinilyticum (0.4< r < 0.7). CONCLUSIONS: These results suggest that the composition of gut microbiota was somewhat affected by the K. japonensis infection. Additionally, increased ratios of pathogenic bacteria (Lawsonia and Plesiomonas) were also associated with the K. japonensis infection, which may therefore increase the likelihood of disease.


Assuntos
Bactérias/classificação , Carpas/microbiologia , Carpas/parasitologia , Infecções por Cestoides/veterinária , Microbioma Gastrointestinal , Intestinos/parasitologia , Animais , Bactérias/isolamento & purificação , Cestoides/fisiologia , Fusobactérias/classificação , Fusobactérias/isolamento & purificação , Proteobactérias/classificação , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S , Tenericutes/classificação , Tenericutes/isolamento & purificação
20.
PLoS One ; 14(5): e0215375, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31050668

RESUMO

This study reports the characterization of the microbial community composition, and the establishment and dynamics of a continuous-flow competitive exclusion culture (CFCEC) derived from gut microbiomes of Nile tilapia (Oreochromis niloticus) specimens reared on aquaculture farms in Colombia. 16S rRNA gene amplicon Illumina sequencing was used to identify taxonomical changes in the CFCEC microbial community over time. The CFCEC was developed from adult tilapia from two farms in Colombia, and CFCEC samples were collected over two months. The pH varied from 6.25 to 6.35 throughout culturing, while anaerobic and aerobic cell counts stabilized at day 9, at 109 CFU mL-1 and were maintained to day 68. A variation in the CFCEC bacterial composition was observed over time. Cetobacterium was the most abundant in the first two days and coincided with a higher CFCEC supernatant antimicrobial effect against the fish pathogen Streptococcus agalactiae. Antimicrobial activity against S. agalactiae disappeared by day 3. Changes in bacterial composition continued to day 33 with Lactococcus spp. becoming the most abundant member of the community. In conclusion, the study of the CFCEC from intestinal tract of Nile tilapia (Oreochromis niloticus) by 16S rRNA gene sequencing allowed identification of predominant bacterial genera in the continuous-flow competitive exclusion culture exhibiting antibacterial activity against the fish pathogen Streptococcus agalactiae.


Assuntos
Bactérias/classificação , Ciclídeos/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Streptococcus agalactiae/crescimento & desenvolvimento , Animais , Bactérias/isolamento & purificação , Carga Bacteriana , Técnicas Bacteriológicas , Colômbia , Fusobactérias/isolamento & purificação , Fusobactérias/fisiologia , Microbioma Gastrointestinal , Lactococcus/isolamento & purificação , Lactococcus/fisiologia , Viabilidade Microbiana , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
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