RESUMEN
Diagnosis and treatment of various diseases in Ayurveda, the Indian system of medicine, relies on 'prakriti' phenotyping of individuals into predominantly three constitutions, kapha, pitta and vata. Recent studies propose that microbiome play an integral role in precision medicine. A study of the relationship between prakriti - the basis of personalized medicine in Ayurveda and that of gut microbiome, and possible biomarker of an individual's health, would vastly improve precision therapy. Towards this, we analyzed bacterial metagenomes from buccal (oral microbiome) and fecal (gut microbiome) samples of 272 healthy individuals of various predominant prakritis. Major bacterial genera from gut microbiome included Prevotella, Bacteroides and Dialister while oral microbiome included Streptococcus, Neisseria, Veilonella, Haemophilus, Porphyromonas and Prevotella. Though the core microbiome was shared across all individuals, we found prakriti specific signatures such as preferential presence of Paraprevotella and Christensenellaceae in vata individuals. A comparison of core gut microbiome of each prakriti with a database of 'healthy' microbes identified microbes unique to each prakriti with functional roles similar to the physiological characteristics of various prakritis as described in Ayurveda. Our findings provide evidence to Ayurvedic interventions based on prakriti phenotyping and possible microbial biomarkers that can stratify the heterogenous population and aid in precision therapy.
Asunto(s)
Medicina Ayurvédica/métodos , Metagenoma , Medicina de Precisión/métodos , Simbiosis/fisiología , Adulto , Técnicas de Tipificación Bacteriana , Bacteroides/clasificación , Bacteroides/genética , Bacteroides/aislamiento & purificación , ADN Bacteriano/genética , Heces/microbiología , Femenino , Microbioma Gastrointestinal/genética , Haemophilus/clasificación , Haemophilus/genética , Haemophilus/aislamiento & purificación , Voluntarios Sanos , Humanos , Masculino , Boca/microbiología , Neisseria/clasificación , Neisseria/genética , Neisseria/aislamiento & purificación , Filogenia , Porphyromonas/clasificación , Porphyromonas/genética , Porphyromonas/aislamiento & purificación , Prevotella/clasificación , Prevotella/genética , Prevotella/aislamiento & purificación , Streptococcus/clasificación , Streptococcus/genética , Streptococcus/aislamiento & purificación , Veillonella/clasificación , Veillonella/genética , Veillonella/aislamiento & purificación , Veillonellaceae/clasificación , Veillonellaceae/genética , Veillonellaceae/aislamiento & purificaciónRESUMEN
Experimental manipulation of gut microbes in animal models alters fear behavior and relevant neurocircuitry. In humans, the first year of life is a key period for brain development, the emergence of fearfulness, and the establishment of the gut microbiome. Variation in the infant gut microbiome has previously been linked to cognitive development, but its relationship with fear behavior and neurocircuitry is unknown. In this pilot study of 34 infants, we find that 1-year gut microbiome composition (Weighted Unifrac; lower abundance of Bacteroides, increased abundance of Veillonella, Dialister, and Clostridiales) is significantly associated with increased fear behavior during a non-social fear paradigm. Infants with increased richness and reduced evenness of the 1-month microbiome also display increased non-social fear. This study indicates associations of the human infant gut microbiome with fear behavior and possible relationships with fear-related brain structures on the basis of a small cohort. As such, it represents an important step in understanding the role of the gut microbiome in the development of human fear behaviors, but requires further validation with a larger number of participants.
Asunto(s)
Bacteroides/genética , Clostridiales/genética , Miedo/psicología , Microbioma Gastrointestinal/genética , Veillonella/genética , Veillonellaceae/genética , Adulto , Bacteroides/clasificación , Bacteroides/aislamiento & purificación , Encéfalo/fisiología , Lactancia Materna , Clostridiales/clasificación , Clostridiales/aislamiento & purificación , Heces/microbiología , Femenino , Humanos , Lactante , Fórmulas Infantiles , Estudios Longitudinales , Masculino , Proyectos Piloto , ARN Ribosómico 16S/genética , Veillonella/clasificación , Veillonella/aislamiento & purificación , Veillonellaceae/clasificación , Veillonellaceae/aislamiento & purificaciónRESUMEN
Strain Marseille-P2082, an anaerobic, non-motile, asporogenous, Gram-negative, coccoid bacterium was isolated from the faeces of a 33 year-old obese French woman before bariatric surgery. The isolate exhibits 98.65% 16S rRNA gene nucleotide sequence similarity with Negativicoccus succinicivorans strain ADV 07/08/06-B-1388T, its current closest phylogenetic neighbour with standing in nomenclature. However, the dDDH relatedness between the new isolate and N. succinicivorans type strain ADV 07/08/06-B-1388T is 52.5 ± 2.7%. Strain Marseille-P2082 has a genome of 1,360,589 bp with a 51.1% G+C content. Its major fatty acids were identified as C18:1n9, C18:0 and C16:0. Based on its phenotypic, genomic and phylogenetic characteristics, strain Marseille-P2082T [= CSURP2082 (Collection de Souches de l'Unité des Rickettsies) = DSM 100853] is proposed as the type strain of the novel species Negativicoccus massiliensis sp. nov. The 16S rRNA gene sequence and whole-genome shotgun sequence have been deposited in EMBL-EBI under accession numbers LN876651 and LT700188, respectively.
Asunto(s)
Microbioma Gastrointestinal , Obesidad , Filogenia , Veillonellaceae/clasificación , Veillonellaceae/aislamiento & purificación , Adulto , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Heces/microbiología , Femenino , Genes Bacterianos/genética , Genoma Bacteriano , Genómica , Humanos , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Veillonellaceae/genética , Veillonellaceae/fisiologíaRESUMEN
An obligately anaerobic, Gram-stain-negative, rod or coccobacilli organism was isolated from a faecal sample of a healthy Japanese woman. In the 16S rRNA gene sequence analysis, strain 5BBH33T showed the highest 16S rRNA gene sequence similarity to Dialister succinatiphilus YIT 11850T (95.9â%), Dialister propionicifaciens ADV 1053.03T (94.3â%), Dialister micraerophilus DSM 19965T (93.1â%), Dialister invisus DSM 15470T (92.5â%) and Dialister pneumosintes ATCC 33048T (91.4â%). The hsp60 gene sequence analysis also revealed strain 5BBH33T had relatively low hsp60 gene sequence similarities (74.4-85.3â%) to other Dialister species. Strain 5BBH33T showed 21.8-23.9â% in silico DNA-DNA hybridization values with other Dialister species. In addition, the average nucleotide identity values between strain 5BBH33T and other Dialister species ranged from 68.7-74.2â%, indicating that this strain should be considered as new species based on whole-genome relatedness. Strain 5BBH33T was asaccharolytic and largely unreactive for commercial kit. However, its growth was enhanced by adding 1â% (w/v) succinate to the medium; strain 5BBH33T was able to decarboxylate succinate to propionate. The strain 5BBH33T genome contained the enzymes involved in succinate utilization. These results improve our understanding of succinate-utilizing bacteria. On the basis of the collected data, strain 5BBH33T represents a novel species in the genus Dialister, for which the name Dialister hominis sp. nov. is proposed. The type strain of D. hominis is 5BBH33T (=JCM 33369T=DSM 109768T).
Asunto(s)
Heces/microbiología , Filogenia , Veillonellaceae/clasificación , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Femenino , Humanos , Japón , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Veillonellaceae/aislamiento & purificaciónRESUMEN
Two strictly anaerobic strains (MB11T and MB56) were isolated from common marmoset (Callithrixjacchus) faeces. Cells of the two strains were Gram-stain-negative, pleomorphic short (strain MB11T) or long (strain MB56) rods. Phylogenetic analysis based on 16S rRNA gene sequences revealed that both isolates were related to the genus Phascolarctobacterium. They had 16S rRNA gene sequences similarities lower than 93â% to previously described species, Phascolarctobacterium faecium ACM 3679T and Phascolarctobacterium succinatutens YIT 12067T, and 98.7â% between themselves. DNA-DNA hybridization values showed that strains MB11T and MB56 were the same species. The genomic DNA G+C content of strains MB11T and MB56 were 47.3-47.4 mol% and 47.7-48.0 mol%. The isolates had different enzymatic activities compared with P. succinatutens JCM 16074T and different major cellular fatty acids compared with P. faecium ACM 3679T. Substrate availability revealed that they utilized not only succinate, but also pyruvate. With pyruvate supplementation, they produced both propionate and acetate, while only propionate production occured with succinate. As suggested by the phylogenic and physiological properties of strains MB11T and MB56, we propose the name Phascolarctobacteriumwakonense sp. nov. with the type strain MB11T (=JCM 32899T=DSM 107697T).
Asunto(s)
Callithrix/microbiología , Heces/microbiología , Filogenia , Veillonellaceae/clasificación , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Japón , Hibridación de Ácido Nucleico , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Veillonellaceae/aislamiento & purificaciónRESUMEN
Dysbiosis, or imbalance in the gut microbiome, has been implicated in auto-immune, inflammatory, neurological diseases as well as in cancers. More recently it has also been shown to be associated with ocular diseases. In the present study, the association of gut microbiome dysbiosis with bacterial Keratitis, an inflammatory eye disease which significantly contributes to corneal blindness, was investigated. Bacterial and fungal gut microbiomes were analysed using fecal samples of healthy controls (HC, n = 21) and bacterial Keratitis patients (BK, n = 19). An increase in abundance of several antiinflammatory organisms including Dialister, Megasphaera, Faecalibacterium, Lachnospira, Ruminococcus and Mitsuokella and members of Firmicutes, Veillonellaceae, Ruminococcaceae and Lachnospiraceae was observed in HC compared to BK patients in the bacterial microbiome. In the fungal microbiome, a decrease in the abundance of Mortierella, Rhizopus, Kluyveromyces, Embellisia and Haematonectria and an increase in the abundance of pathogenic fungi Aspergillus and Malassezia were observed in BK patients compared to HC. In addition, heatmaps, PCoA plots and inferred functional profiles also indicated significant variations between the HC and BK microbiomes, which strongly suggest dysbiosis in the gut microbiome of BK patients. This is the first study demonstrating the association of gut microbiome with the pathophysiology of BK and thus supports the gut-eye axis hypothesis. Considering that Keratitis affects about 1 million people annually across the globe, the data could be the basis for developing alternate strategies for treatment like use of probiotics or fecal transplantation to restore the healthy microbiome as a treatment protocol for Keratitis.
Asunto(s)
ADN Bacteriano/genética , ADN de Hongos/genética , Disbiosis/microbiología , Microbioma Gastrointestinal/fisiología , Queratitis/microbiología , Adulto , Aspergillus/clasificación , Aspergillus/genética , Aspergillus/aislamiento & purificación , Estudios de Casos y Controles , Clostridiales/clasificación , Clostridiales/genética , Clostridiales/aislamiento & purificación , ADN Bacteriano/aislamiento & purificación , ADN de Hongos/aislamiento & purificación , Disbiosis/diagnóstico , Disbiosis/patología , Faecalibacterium/clasificación , Faecalibacterium/genética , Faecalibacterium/aislamiento & purificación , Heces/microbiología , Femenino , Humanos , Queratitis/diagnóstico , Queratitis/patología , Kluyveromyces/clasificación , Kluyveromyces/genética , Kluyveromyces/aislamiento & purificación , Malassezia/clasificación , Malassezia/genética , Malassezia/aislamiento & purificación , Masculino , Megasphaera/clasificación , Megasphaera/genética , Megasphaera/aislamiento & purificación , Persona de Mediana Edad , Mortierella/clasificación , Mortierella/genética , Mortierella/aislamiento & purificación , Rhizopus/clasificación , Rhizopus/genética , Rhizopus/aislamiento & purificación , Ruminococcus/clasificación , Ruminococcus/genética , Ruminococcus/aislamiento & purificación , Veillonellaceae/clasificación , Veillonellaceae/genética , Veillonellaceae/aislamiento & purificaciónRESUMEN
BACKGROUND: Gut microbiota plays an important role in many metabolic diseases such as diabetes and atherosclerosis. Apolipoprotein E (apoE) knock-out (KO) mice are frequently used for the study of hyperlipidemia and atherosclerosis. However, it is unknown whether apoE KO mice have altered gut microbiota when challenged with a Western diet. METHODS: In the current study, we assessed the gut microbiota profiling of apoE KO mice and compared with wild-type mice fed either a normal chow or Western diet for 12 weeks using 16S pyrosequencing. RESULTS: On a western diet, the gut microbiota diversity was significantly decreased in apoE KO mice compared with wild type (WT) mice. Firmicutes and Erysipelotrichaceae were significantly increased in WT mice but Erysipelotrichaceae was unchanged in apoE KO mice on a Western diet. The weighted UniFrac principal coordinate analysis exhibited clear separation between WT and apoE KO mice on the first vector (58.6%) with significant changes of two dominant phyla (Bacteroidetes and Firmicutes) and seven dominant families (Porphyromonadaceae, Lachnospiraceae, Ruminococcaceae, Desulfovibrionaceae, Helicobacteraceae, Erysipelotrichaceae and Veillonellaceae). Lachnospiraceae was significantly enriched in apoE KO mice on a Western diet. In addition, Lachnospiraceae and Ruminococcaceae were positively correlated with relative atherosclerosis lesion size in apoE KO. CONCLUSIONS: Collectively, our study showed that there are marked changes in the gut microbiota of apoE KO mice, particularly challenged with a Western diet and these alterations may be possibly associated with atherosclerosis.
Asunto(s)
Apolipoproteínas E/deficiencia , Aterosclerosis/microbiología , Dieta Occidental/efectos adversos , Microbioma Gastrointestinal/genética , Hiperlipidemias/microbiología , Animales , Apolipoproteínas E/genética , Aterosclerosis/etiología , Aterosclerosis/genética , Aterosclerosis/patología , Bacteroidetes/clasificación , Bacteroidetes/genética , Bacteroidetes/aislamiento & purificación , ADN Ribosómico/genética , Desulfovibrionaceae/clasificación , Desulfovibrionaceae/genética , Desulfovibrionaceae/aislamiento & purificación , Modelos Animales de Enfermedad , Firmicutes/clasificación , Firmicutes/genética , Firmicutes/aislamiento & purificación , Helicobacteraceae/clasificación , Helicobacteraceae/genética , Helicobacteraceae/aislamiento & purificación , Hiperlipidemias/etiología , Hiperlipidemias/genética , Hiperlipidemias/patología , Lípidos/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados para ApoE , Porphyromonas/clasificación , Porphyromonas/genética , Porphyromonas/aislamiento & purificación , ARN Ribosómico 16S/genética , Ruminococcus/clasificación , Ruminococcus/genética , Ruminococcus/aislamiento & purificación , Análisis de Secuencia de ADN , Índice de Severidad de la Enfermedad , Veillonellaceae/clasificación , Veillonellaceae/genética , Veillonellaceae/aislamiento & purificaciónRESUMEN
BACKGROUND: Almond processing has been shown to differentially impact metabolizable energy; however, the effect of food form on the gastrointestinal microbiota is under-investigated. OBJECTIVE: We aimed to assess the interrelationship of almond consumption and processing on the gastrointestinal microbiota. DESIGN: A controlled-feeding, randomized, five-period, crossover study with washouts between diet periods was conducted in healthy adults (n = 18). Treatments included: (1) zero servings/day of almonds (control); (2) 1.5 servings (42 g)/day of whole almonds; (3) 1.5 servings/day of whole, roasted almonds; (4) 1.5 servings/day of roasted, chopped almonds; and (5) 1.5 servings/day of almond butter. Fecal samples were collected at the end of each three-week diet period. RESULTS: Almond consumption increased the relative abundances of Lachnospira, Roseburia, and Dialister (p ≤ 0.05). Comparisons between control and the four almond treatments revealed that chopped almonds increased Lachnospira, Roseburia, and Oscillospira compared to control (p < 0.05), while whole almonds increased Dialister compared to control (p = 0.007). There were no differences between almond butter and control. CONCLUSIONS: These results reveal that almond consumption induced changes in the microbial community composition of the human gastrointestinal microbiota. Furthermore, the degree of almond processing (e.g., roasting, chopping, and grinding into butter) differentially impacted the relative abundances of bacterial genera.
Asunto(s)
Disbiosis/prevención & control , Alimentos Funcionales , Microbioma Gastrointestinal , Nueces , Prunus dulcis , Anciano , Clostridiales/clasificación , Clostridiales/crecimiento & desarrollo , Clostridiales/aislamiento & purificación , Condimentos , Culinaria , Estudios Cruzados , Disbiosis/microbiología , Heces/microbiología , Femenino , Estudios de Seguimiento , Manipulación de Alimentos , Humanos , Masculino , Persona de Mediana Edad , Tipificación Molecular , Veillonellaceae/clasificación , Veillonellaceae/crecimiento & desarrollo , Veillonellaceae/aislamiento & purificaciónRESUMEN
To provide new insight into the dark fermentation process, a multi-lateral study was performed to study the microbiology of 20 different lab-scale bioreactors operated in four different countries (Brazil, Chile, Mexico, and Uruguay). Samples (29) were collected from bioreactors with different configurations, operation conditions, and performances. The microbial communities were analyzed using 16S rRNA genes 454 pyrosequencing. The results showed notably uneven communities with a high predominance of a particular genus. The phylum Firmicutes predominated in most of the samples, but the phyla Thermotogae or Proteobacteria dominated in a few samples. Genera from three physiological groups were detected: high-yield hydrogen producers (Clostridium, Kosmotoga, Enterobacter), fermenters with low-hydrogen yield (mostly from Veillonelaceae), and competitors (Lactobacillus). Inocula, reactor configurations, and substrates influence the microbial communities. This is the first joint effort that evaluates hydrogen-producing reactors and operational conditions from different countries and contributes to understand the dark fermentation process.
Asunto(s)
Reactores Biológicos/normas , Fermentación , Hidrógeno/metabolismo , Consorcios Microbianos/genética , ARN Ribosómico 16S/genética , Anaerobiosis , Clostridium/clasificación , Clostridium/genética , Clostridium/metabolismo , Enterobacter/clasificación , Enterobacter/genética , Enterobacter/metabolismo , Firmicutes/clasificación , Firmicutes/genética , Firmicutes/metabolismo , Secuenciación de Nucleótidos de Alto Rendimiento , Lactobacillus/clasificación , Lactobacillus/genética , Lactobacillus/metabolismo , América Latina , Proteobacteria/clasificación , Proteobacteria/genética , Proteobacteria/metabolismo , Thermotoga maritima/clasificación , Thermotoga maritima/genética , Thermotoga maritima/metabolismo , Veillonellaceae/clasificación , Veillonellaceae/genética , Veillonellaceae/metabolismoRESUMEN
Recurrent aphthous stomatitis (RAS) is the most common disease affecting oral mucosae. Etiology is unknown, but several factors have been implicated, all of which influence the composition of microbiota residing on oral mucosae, which in turn modulates immunity and thereby affects disease progression. Although no individual pathogens have been conclusively shown to be causative agents of RAS, imbalanced composition of the oral microbiota may play a key role. In this study, we sought to determine composition profiles of bacterial microbiota in the oral mucosa associated with RAS. Using high-throughput 16S rRNA gene sequencing, we characterized the most abundant bacterial populations residing on healthy and ulcerated mucosae in patients with RAS (recruited using highly stringent criteria) and no associated medical conditions; we also compared these to the bacterial microbiota of healthy controls (HCs). Phylum-level diversity comparisons revealed decreased Firmicutes and increased Proteobacteria in ulcerated sites, as compared with healthy sites in RAS patients, and no differences between RAS patients with healthy sites and HCs. Genus-level analysis demonstrated higher abundance of total Bacteroidales in RAS patients with healthy sites over HCs. Porphyromonadaceae comprising species associated with periodontal disease and Veillonellaceae predominated in ulcerated sites over HCs, while no quantitative differences of these families were observed between healthy sites in RAS patients and HCs. Streptococcaceae comprising species associated with oral health predominated in HCs over ulcerated sites but not in HCs over healthy sites in RAS patients. This study demonstrates that mucosal microbiome changes in patients with idiopathic RAS--namely, increased Bacteroidales species in mucosae of RAS patients not affected by active ulceration. While these changes suggest a microbial role in initiation of RAS, this study does not provide data on causality. Within this limitation, the study contributes to the understanding of the potential role of mucosal microbiome changes in oral mucosal disease.
Asunto(s)
Microbiota , Mucosa Bucal/microbiología , Estomatitis Aftosa/microbiología , Adolescente , Adulto , Bacterias/clasificación , Fenómenos Fisiológicos Bacterianos , Bacteroidaceae/clasificación , Estudios de Casos y Controles , Femenino , Bacterias Gramnegativas/clasificación , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Persona de Mediana Edad , Porphyromonas/clasificación , Proteobacteria/clasificación , Recurrencia , Streptococcaceae/clasificación , Veillonellaceae/clasificación , Adulto JovenRESUMEN
Little is known about the virulence and clinical impact on humans from infection with Anaeroglobus geminates, an anaerobic gram-negative coccus belonging to the family Veillonellaceae. We report the first case of an Anaeroglobus geminates invasive infection in humans characterized by pneumonia complicated with empyema. The pathogen was initially identified as Veillonella spp. by an automatic identification system (Becton-Dickinson and Company, Franklin Lakes, NJ, USA) and definitively identified following 16S ribosomal RNA gene sequence analysis. The patient was cured by surgical decortication and antimicrobial therapy. In this case, the combination of effective antibiotics, surgical intervention, and adequate drainage successfully cured the patient.
Asunto(s)
Empiema , Infecciones por Bacterias Gramnegativas , Neumonía Bacteriana , Veillonellaceae , Anciano , ADN Bacteriano/análisis , ADN Bacteriano/genética , Femenino , Humanos , Radiografía Torácica , Veillonellaceae/clasificación , Veillonellaceae/genéticaRESUMEN
A strictly anaerobic bacterial strain, WK011(T), was isolated from a methanogenic reactor treating waste from cattle farms. The cells stained Gram-negative and were curved rods with a polar or subpolar flagellum. Spore formation was not observed. The optimum temperature for growth was 35 °C and the optimum pH was 6.7. Tests for oxidase, catalase and nitrate-reduction activities were negative. Hydrogen sulfide was produced. The strain fermented carbohydrates and produced acetate and propionate as major fermentation products. The genomic DNA G+C content was 41.7 mol%. The major cellular fatty acids were C15:0, C16:1ω9c and C18:1 dimethylacetal. The diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid. The most closely related species to strain WK011(T) on the basis of 16S rRNA gene sequences were Propionispira arboris and Zymophilus raffinosivorans (95.6% sequence similarity to the type strains of both species). It was shown by phylogenetic and phenotypic examination of the type strains of related species, including the second species of the genus Zymophilus, Zymophilus paucivorans, that the two genera should be combined and that the two species of the genus Zymophilus should be transferred to the genus Propionispira, as Propionispira raffinosivorans comb. nov. (type strain SH2(T) = ATCC 49691(T)â=âDSM 20765(T)) and Propionispira paucivorans comb. nov. (type strain AA1(T) = ATCC 49689(T) = DSM 20756(T)), with an emended description of the genus Propionispira. Based on differences in the phylogenetic and phenotypic characteristics of strain WK011(T) from those of closely related species, the novel species Propionispira arcuata sp. nov. is proposed to accommodate the strain. The type strain is WK011(T) ( = JCM 16475(T) = DSM 22929(T)).
Asunto(s)
Reactores Biológicos/microbiología , Filogenia , Veillonellaceae/clasificación , Animales , Bacterias Anaerobias/clasificación , Bacterias Anaerobias/genética , Bacterias Anaerobias/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Composición de Base , Bovinos , ADN Bacteriano/genética , Ácidos Grasos/química , Fermentación , Japón , Datos de Secuencia Molecular , Peptidoglicano/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Veillonellaceae/genética , Veillonellaceae/aislamiento & purificaciónAsunto(s)
Isópteros/microbiología , Veillonellaceae/clasificación , Veillonellaceae/aislamiento & purificación , Animales , Técnicas de Tipificación Bacteriana , Análisis por Conglomerados , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Tracto Gastrointestinal/microbiología , Microscopía , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Veillonellaceae/genética , Veillonellaceae/metabolismoRESUMEN
Following the isolation, cultivation and characterization of the rumen bacterium Anaerovibrio lipolyticus in the 1960s, it has been recognized as one of the major species involved in lipid hydrolysis in ruminant animals. However, there has been limited characterization of the lipases from the bacterium, despite the importance of understanding lipolysis and its impact on subsequent biohydrogenation of polyunsaturated fatty acids by rumen microbes. This study describes the draft genome of Anaerovibrio lipolytica 5ST, and the characterization of three lipolytic genes and their translated protein. The uncompleted draft genome was 2.83 Mbp and comprised of 2,673 coding sequences with a G+C content of 43.3%. Three putative lipase genes, alipA, alipB and alipC, encoding 492-, 438- and 248- amino acid peptides respectively, were identified using RAST. Phylogenetic analysis indicated that alipA and alipB clustered with the GDSL/SGNH family II, and alipC clustered with lipolytic enzymes from family V. Subsequent expression and purification of the enzymes showed that they were thermally unstable and had higher activities at neutral to alkaline pH. Substrate specificity assays indicated that the enzymes had higher hydrolytic activity against caprylate (C8), laurate (C12) and myristate (C14).
Asunto(s)
Lipasa/genética , Lipasa/metabolismo , Veillonellaceae/enzimología , Veillonellaceae/genética , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Secuencia Conservada , Activación Enzimática , Expresión Génica , Concentración de Iones de Hidrógeno , Lipasa/química , Lipasa/aislamiento & purificación , Lipólisis , Datos de Secuencia Molecular , Filogenia , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Especificidad por Sustrato , Temperatura , Veillonellaceae/clasificaciónRESUMEN
Pelosinus spp. are fermentative firmicutes that were recently reported to be prominent members of microbial communities at contaminated subsurface sites in multiple locations. Here we report metabolic characteristics and their putative genetic basis in Pelosinus sp. strain HCF1, an isolate that predominated anaerobic, Cr(VI)-reducing columns constructed with aquifer sediment. Strain HCF1 ferments lactate to propionate and acetate (the methylmalonyl-coenzyme A [CoA] pathway was identified in the genome), and its genome encodes two [NiFe]- and four [FeFe]-hydrogenases for H(2) cycling. The reduction of Cr(VI) and Fe(III) may be catalyzed by a flavoprotein with 42 to 51% sequence identity to both ChrR and FerB. This bacterium has unexpected capabilities and gene content associated with reduction of nitrogen oxides, including dissimilatory reduction of nitrate to ammonium (two copies of NrfH and NrfA were identified along with NarGHI) and a nitric oxide reductase (NorCB). In this strain, either H(2) or lactate can act as a sole electron donor for nitrate, Cr(VI), and Fe(III) reduction. Transcriptional studies demonstrated differential expression of hydrogenases and nitrate and nitrite reductases. Overall, the unexpected metabolic capabilities and gene content reported here broaden our perspective on what biogeochemical and ecological roles this species might play as a prominent member of microbial communities in subsurface environments.
Asunto(s)
Cromatos/metabolismo , ADN Bacteriano/genética , Genoma Bacteriano , Veillonellaceae/genética , Veillonellaceae/metabolismo , Ácido Acético/metabolismo , ADN Bacteriano/química , Fermentación , Flavoproteínas/genética , Agua Subterránea/microbiología , Hierro/metabolismo , Ácido Láctico/metabolismo , Redes y Vías Metabólicas/genética , Datos de Secuencia Molecular , Nitratos/metabolismo , Oxidación-Reducción , Propionatos/metabolismo , Análisis de Secuencia de ADN , Veillonellaceae/clasificación , Veillonellaceae/aislamiento & purificaciónRESUMEN
BACKGROUND: To understand the relationship between our bacterial microbiome and health, it is essential to define the microbiome in the absence of disease. The digestive tract includes diverse habitats and hosts the human body's greatest bacterial density. We describe the bacterial community composition of ten digestive tract sites from more than 200 normal adults enrolled in the Human Microbiome Project, and metagenomically determined metabolic potentials of four representative sites. RESULTS: The microbiota of these diverse habitats formed four groups based on similar community compositions: buccal mucosa, keratinized gingiva, hard palate; saliva, tongue, tonsils, throat; sub- and supra-gingival plaques; and stool. Phyla initially identified from environmental samples were detected throughout this population, primarily TM7, SR1, and Synergistetes. Genera with pathogenic members were well-represented among this disease-free cohort. Tooth-associated communities were distinct, but not entirely dissimilar, from other oral surfaces. The Porphyromonadaceae, Veillonellaceae and Lachnospiraceae families were common to all sites, but the distributions of their genera varied significantly. Most metabolic processes were distributed widely throughout the digestive tract microbiota, with variations in metagenomic abundance between body habitats. These included shifts in sugar transporter types between the supragingival plaque, other oral surfaces, and stool; hydrogen and hydrogen sulfide production were also differentially distributed. CONCLUSIONS: The microbiomes of ten digestive tract sites separated into four types based on composition. A core set of metabolic pathways was present across these diverse digestive tract habitats. These data provide a critical baseline for future studies investigating local and systemic diseases affecting human health.
Asunto(s)
Biota , Heces/microbiología , Metagenoma , Boca/microbiología , Tonsila Palatina/microbiología , Faringe/microbiología , Actinobacteria/clasificación , Actinobacteria/genética , Actinobacteria/aislamiento & purificación , Adolescente , Adulto , Técnicas de Tipificación Bacteriana/métodos , Bacteroidetes/clasificación , Bacteroidetes/genética , Bacteroidetes/aislamiento & purificación , Femenino , Genes de ARNr , Humanos , Masculino , Filogenia , ARN Ribosómico 16S/análisis , ARN Ribosómico 16S/genética , Saliva/microbiología , Veillonellaceae/clasificación , Veillonellaceae/genética , Veillonellaceae/aislamiento & purificación , Adulto JovenRESUMEN
Salmonella continues to be a significant human health threat, and the objective of this study was to identify microorganisms with the potential to improve porcine food-safety through their antagonism of Salmonella. Anaerobic culture supernatants of 973 bacterial isolates from the gastrointestinal tract and feces of swine were screened for their capacity to inhibit the growth of Salmonella enterica serovar Typhimurium. Growth inhibition of 1000-fold or greater was observed from 16 isolates, and 16S rRNA sequencing identified the isolates as members of the genera Mitsuokella, Escherichia/Shigella, Anaerovibrio, Selenomonas, and Streptococcus. Four isolates were identified as Mitsuokella jalaludinii, and the mechanism of Salmonella Typhimurium growth inhibition by M. jalaludinii was further investigated. M. jalaludinii stationary phase culture supernatants were observed to significantly inhibit growth, and featured the production of lactic, succinic, and acetic acids. Aerobic and anaerobic S. Typhimurium growth was restored when the pH of the culture supernatants (pH 4.6) was increased to pH 6.8. However, S. Typhimurium growth in fermentation acid-free media was the same at pH 4.6 and pH 6.8 - indicating a synergistic effect between fermentation acid production and low pH as the cause of S. Typhimurium growth inhibition. Furthermore, exposure of S. Typhimurium to M. jalaludinii culture supernatants inhibited Salmonella invasion of HEp-2 cells by 10-fold. The results identify M. jalaludinii as a possible inhibitor of Salmonella growth and invasion in swine, and thus a potential probiotic capable of improving food safety.
Asunto(s)
Antibiosis , Veillonellaceae/fisiología , Animales , Bacterias/genética , Bacterias/aislamiento & purificación , Línea Celular , Medios de Cultivo/química , Heces/microbiología , Tracto Gastrointestinal/microbiología , Humanos , Concentración de Iones de Hidrógeno , Filogenia , ARN Ribosómico 16S/genética , Salmonella typhimurium/crecimiento & desarrollo , Porcinos/microbiología , Veillonellaceae/clasificación , Veillonellaceae/genéticaRESUMEN
The determination of the success of in situ bioremediation strategies is complex. By using controlled laboratory conditions, the influence of individual variables, such as U(VI), Cr(VI), and electron donors and acceptors on community structure, dynamics, and the metal-reducing potential can be studied. Triplicate anaerobic, continuous-flow reactors were inoculated with Cr(VI)-contaminated groundwater from the Hanford, WA, 100-H area, amended with lactate, and incubated for 95 days to obtain stable, enriched communities. The reactors were kept anaerobic with N(2) gas (9 ml/min) flushing the headspace and were fed a defined medium amended with 30 mM lactate and 0.05 mM sulfate with a 48-h generation time. The resultant diversity decreased from 63 genera within 12 phyla to 11 bacterial genera (from 3 phyla) and 2 archaeal genera (from 1 phylum). Final communities were dominated by Pelosinus spp. and to a lesser degree, Acetobacterium spp., with low levels of other organisms, including methanogens. Four new strains of Pelosinus were isolated, with 3 strains being capable of Cr(VI) reduction while one also reduced U(VI). Under limited sulfate, it appeared that the sulfate reducers, including Desulfovibrio spp., were outcompeted. These results suggest that during times of electron acceptor limitation in situ, organisms such as Pelosinus spp. may outcompete the more-well-studied organisms while maintaining overall metal reduction rates and extents. Finally, lab-scale simulations can test new strategies on a smaller scale while facilitating community member isolation, so that a deeper understanding of community metabolism can be revealed.
Asunto(s)
Biodegradación Ambiental , Ecosistema , Sedimentos Geológicos/microbiología , Lactatos/metabolismo , Uranio/metabolismo , Veillonellaceae/crecimiento & desarrollo , Archaea/clasificación , Archaea/genética , Archaea/crecimiento & desarrollo , Archaea/aislamiento & purificación , Archaea/metabolismo , Bacterias/clasificación , Bacterias/genética , Bacterias/crecimiento & desarrollo , Bacterias/aislamiento & purificación , Bacterias/metabolismo , Reactores Biológicos , Cromo/metabolismo , Medios de Cultivo , ADN de Archaea/análisis , ADN de Archaea/genética , ADN Bacteriano/análisis , ADN Bacteriano/genética , Agua Subterránea/microbiología , Datos de Secuencia Molecular , Oxidación-Reducción , Filogenia , ARN Ribosómico 16S , Análisis de Secuencia de ADN , Veillonellaceae/clasificación , Veillonellaceae/genética , Veillonellaceae/aislamiento & purificaciónRESUMEN
Two anaerobic bacterial strains, designated SHI-1(T) and SHI-2, were isolated from chlorinated solvent-contaminated groundwater. They were found to be identical in phenotypic properties and shared high (98.5-99.8 %) pairwise 16S rRNA gene sequence similarity. Multiple 16S rRNA genes were found to be present in the isolates as well as Pelosinus fermentans DSM 17108(T) and Sporotalea propionica DSM 13327(T). Strains SHI-1(T) and SHI-2 could be differentiated from their closest phylogenetic relatives, P. fermentans DSM 17108(T) and S. propionica DSM 13327(T), on the basis of their phenotypic and phylogenetic properties. The isolates were Gram-negative, spore-forming, motile rods with peritrichous flagella. Growth occurred at 10-42 °C and pH 5.5-8.5. Fermentative growth was observed on Casamino acids, fructose, fumarate, glucose, glycerol, pyruvate and yeast extract. The major organic acids produced from glucose and glycerol fermentation were propionate and acetate. The major organic acids produced from fermentation of fumarate were propionate, acetate and succinate. The major cellular fatty acids were summed feature 4 (consisting of C(15:1)ω8c and/or C(15:2)), summed feature 8 (consisting of C(17:1)ω8c and/or C(17:2)) and C(14:0) dimethyl aldehyde. The polar lipids comprised aminophospholipids, including phosphatidylethanolamine and phosphatidylserine, and an unknown phospholipid. The genomic DNA G+C content was 39.2 mol%. We propose that strains SHI-1(T) and SHI-2 are assigned to a novel species of the genus Pelosinus, with the name Pelosinus defluvii sp. nov. (type strain SHI-1(T) = NRRL Y-59407(T) = LMG 25549(T)). The description of the genus Pelosinus is emended. We also propose the transfer of S. propionica to the genus Pelosinus as Pelosinus propionicus comb. nov. (type strain TmPN3(T) = DSM 13327(T) = ATCC BAA-626(T)), on the basis of phylogenetic, chemotaxonomic and phenotypic properties.
Asunto(s)
Agua Subterránea/microbiología , Veillonellaceae/clasificación , Veillonellaceae/aislamiento & purificación , Composición de Base , ADN Bacteriano/genética , Agua Subterránea/análisis , Halogenación , Datos de Secuencia Molecular , Filogenia , ARN Ribosómico 16S/genética , Veillonellaceae/genética , Veillonellaceae/metabolismo , Contaminación Química del Agua/análisisRESUMEN
Isolation, cultivation, and characterization of the intestinal microorganisms are important for understanding the comprehensive physiology of the human gastrointestinal (GI) tract microbiota. Here, we isolated two novel bacterial strains, YIT 12067(T) and YIT 12068, from the feces of healthy human adults. Phylogenetic analysis indicated that they belonged to the same species and were most closely related to Phascolarctobacterium faecium ACM 3679(T), with 91.4% to 91.5% 16S rRNA gene sequence similarities, respectively. Substrate availability tests revealed that the isolates used only succinate; they did not ferment any other short-chain fatty acids or carbohydrates tested. When these strains were cocultured with the xylan-utilizing and succinate-producing bacterium Paraprevotella xylaniphila YIT 11841(T), in medium supplemented with xylan but not succinate, their cell numbers became 2 to 3 orders of magnitude higher than those of the monoculture; succinate became undetectable, and propionate was formed. Database analysis revealed that over 200 uncultured bacterial clones from the feces of humans and other mammals showed high sequence identity (>98.7%) to YIT 12067(T). Real-time PCR analysis also revealed that YIT 12067(T)-like bacteria were present in 21% of human fecal samples, at an average level of 3.34 × 10(8) cells/g feces. These results indicate that YIT 12067(T)-like bacteria are distributed broadly in the GI tract as subdominant members that may adapt to the intestinal environment by specializing to utilize the succinate generated by other bacterial species. The phylogenetic and physiological properties of YIT 12067(T) and YIT 12068 suggest that these strains represent a novel species, which we have designated Phascolarctobacterium succinatutens sp. nov.