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1.
Mater Today Bio ; 15: 100275, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35572854

RESUMO

Peri-implantitis, the major cause of implant failure, is an inflammatory destructive disease due to the dysbiotic polymicrobial communities at the peri-implant sites. Therefore, it is highly warranted to develop the implant materials with antimicrobial properties and investigate their effects on oral microbiota. However, most of the relevant studies were performed in vitro, and insufficient to provide the comprehensive assessment of the antimicrobial capacity of the implant materials in vivo. Herein, we introduce an innovative approach to evaluate the in vivo antibacterial properties of the most commonly used implant materials, titanium with different nanostructured surfaces, and investigate their antibacterial mechanism via the next-generation sequencing (NGS) technology. We firstly prepared the titanium implants with three different surfaces, i) mechanical polishing (MP), ii) TiO2 nanotubes (NT) and iii) nanophase calcium phosphate embedded to TiO2 nanotubes (NTN), and then characterized them using scanning electron microscopy (SEM), energy-dispersive X-ray spectrometer (EDS), X-ray photoelectron spectroscopy (XPS), confocal laser scanning microscopy (CLSM) and surface hydrophilicity analysis. Afterwards, the implants were placed in the beagle dogs' mouths to replace the pre-extracted premolar and molar teeth for eight weeks through implant surgery. The supra- and sub-mucosal plaques were collected and subjected to 16S rRNA gene/RNA sequencing and data analysis. It was found that the nanostructured surfaces in NT and NTN groups showed significantly increased roughness and decreased water contact angles compared to the MP group, while the XPS data further confirmed the successful modifications of TiO2 nanotubes and the subsequent deposition of nanophase calcium phosphate. Notably, the nanostructured surfaces in NT and NTN groups had limited impact on the diversity and community structure of oral microbiota according to the 16S rRNA sequencing results, and the nanostructures in NTN group could down-regulate the genes associated with localization and locomotion based on Gene Ontology (GO) terms enrichment analysis. Moreover, the differentially expressed genes (DEGs) were associated with microbial metabolism, protein synthesis and bacterial invasion of epithelial cells. Taken together, this study provides a new strategy to evaluate the antibacterial properties of the biomedical materials in vivo via the high-throughput sequencing and bioinformatic approaches, revealing the differences of the composition and functional gene expressions in the supra- and sub-mucosal microbiome.

2.
Microbiol Spectr ; 9(2): e0070121, 2021 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-34585987

RESUMO

More than 75 species/species-level phylotypes belonging to the genus Treponema inhabit the human oral cavity. Treponema denticola is commonly associated with periodontal disease, but the etiological roles and ecological distributions of other oral treponemes remain more obscure. Here, we compared the clinical distributions of phylogroup 1 and 2 oral treponemes in subgingival plaque sampled from Chinese subjects with periodontitis (n = 10) and gingivitis (n = 8) via sequence analysis of the highly conserved pyrH housekeeping gene. Two PCR primer sets that targeted oral phylogroup 1 and 2 treponeme pyrH genes were used to construct plasmid clone amplicon libraries for each subject, and the libraries were sequenced for bioinformatic analysis. A total of 1,204 quality-filtered, full-length pyrH gene sequences were obtained from the cohort (median number, 61.5 cloned pyrH sequences per subject; range, 59 to 83), which were assigned to 34 pyrH genotypes (designated pyrH001 to pyrH034; 97% sequence identity cutoff). Eighteen pyrH genotypes (536 pyrH sequences) corresponded to phylogroup 1 treponeme taxa (including Treponema vincentii and Treponema medium). Sixteen pyrH genotypes (668 pyrH sequences) corresponded to T. denticola and other phylogroup 2 treponemes. Samples from periodontitis subjects contained a greater diversity of phylogroup 2 pyrH genotypes than did samples from gingivitis subjects (Mann-Whitney U test). One T. denticola pyrH genotype (pyrH001) was highly prevalent, detected in 10/10 periodontitis and 6/8 gingivitis subjects. Several subjects harbored multiple T. denticola pyrH genotypes. Nonmetric multidimensional scaling and permutational multivariate analysis of variance (PERMANOVA) revealed no significant differences in overall pyrH genotype compositions between periodontitis and gingivitis subjects. Taken together, our results show that subjects with periodontitis and gingivitis commonly harbor highly taxonomically diverse communities of oral treponemes. IMPORTANCE Periodontal diseases, such as periodontitis, are highly complex, multifactorial inflammatory infectious diseases affecting the gums and tooth-supporting structures. They are caused by chronic accumulations of dental plaque below the gum line that typically comprise hundreds of different bacterial species. Certain species of spiral-shaped bacteria known as treponemes, most notably Treponema denticola, are proposed to play key roles in the development and progression of periodontal disease. In our study, we characterized the genetic lineages of T. denticola, Treponema vincentii, Treponema medium, and related species of treponeme bacteria that were present in dental plaque samples from Chinese subjects with periodontal disease. Our results revealed that individual subjects commonly harbored multiple genetic lineages (strains) of T. denticola and other species of treponeme bacteria. Taken together, our results indicate that highly diverse and complex populations of oral treponemes may be present in dental plaque, which may potentially play important roles affecting periodontal health status.


Assuntos
Gengivite/microbiologia , Periodontite/microbiologia , Filogenia , Treponema denticola/classificação , Treponema/classificação , Infecções por Treponema/microbiologia , Estudos de Coortes , Microbiota , Boca/microbiologia , Doenças Periodontais/microbiologia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Especificidade da Espécie , Treponema/genética , Treponema denticola/genética
3.
Microbiol Resour Announc ; 9(22)2020 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-32467286

RESUMO

Host-associated treponeme bacteria play etiological roles in human and animal soft tissue infections. Treponema sp. strain OMZ 804 (ATCC 700766) was isolated from dental plaque sampled from a patient with periodontitis in Switzerland in 1994. We report here the complete genome sequence of its 2.98-Mb circular chromosome.

4.
J Bacteriol ; 202(12)2020 05 27.
Artigo em Inglês | MEDLINE | ID: mdl-32229530

RESUMO

Bacteriocins are ribosomally synthesized proteinaceous antibacterial peptides. They selectively interfere with the growth of other bacteria. The production and secretion of bacteriocins confer a distinct ecological advantage to the producer in competing against other bacteria that are present in the same ecological niche. Streptococcus mutans, a significant contributor to the development of dental caries, is one of the most prolific producers of bacteriocins, known as mutacins in S. mutans In this study, we characterized the locus encoding mutacin B-Ny266, a lantibiotic with a broad spectrum of activity. The chromosomal locus is composed of six predicted operon structures encoding proteins involved in regulation, antimicrobial activity, biosynthesis, modification, transport, and immunity. Mutacin B-Ny266 was purified from semisolid cultures, and two inhibitory peptides, LanA and LanA', were detected. Both peptides were highly modified. Such modifications include dehydration of serine and threonine and the formation of a C-terminal aminovinyl-cysteine (AviCys) ring. While LanA peptide alone is absolutely required for antimicrobial activity, the presence of LanA' enhanced the activity of LanA, suggesting that B-Ny266 may function as a two-peptide lantibiotic. The activation of lanAA' expression is most likely controlled by the conserved two-component system NsrRS, which is activated by LanA peptide but not by LanA'. The chromosomal locus encoding mutacin B-Ny266 was not universally conserved in all sequenced S. mutans genomes. Intriguingly, the genes encoding LanAA' peptides were restricted to the most invasive serotypes of S. mutansIMPORTANCE Although dental caries is largely preventable, it remains the most common and costly infectious disease worldwide. Caries is initiated by the presence of dental plaque biofilm that contains Streptococcus mutans, a species extensively characterized by its role in caries development and formation. S. mutans deploys an arsenal of strategies to establish itself within the oral cavity. One of them is the production of bacteriocins that confer a competitive advantage by targeting and killing closely related competitors. In this work, we found that mutacin B-Ny266 is a potent lantibiotic that is effective at killing a wide array of oral streptococci, including nearly all S. mutans strains tested. Lantibiotics produced by oral bacteria could represent a promising strategy to target caries pathogens embedded in dental plaque biofilm.


Assuntos
Antibacterianos/biossíntese , Proteínas de Bactérias/genética , Bacteriocinas/biossíntese , Cárie Dentária/microbiologia , Streptococcus mutans/genética , Streptococcus mutans/metabolismo , Antibacterianos/farmacologia , Proteínas de Bactérias/metabolismo , Bacteriocinas/farmacologia , Genoma Bacteriano , Humanos , Óperon , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/crescimento & desenvolvimento
5.
Microbiol Resour Announc ; 9(1)2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31896633

RESUMO

Amplicon sequencing of the 16S rRNA gene is commonly performed for the assessment and comparison of microbiomes. Here, we introduce GenePiper, an open-source R Shiny application that provides an easy-to-use interface, a wide range of analytical methods, and optimized graphical outputs for offline microbiome data analyses.

6.
Artigo em Inglês | MEDLINE | ID: mdl-31681625

RESUMO

Objective: To profile the salivary microbiomes of a Hong Kong Chinese cohort at a species-level resolution and determine species that discriminated clinically resolved periodontitis from periodontally healthy cases. Methods: Salivary microbiomes of 35 Hong Kong Chinese subjects' under routine supportive dental care were analyzed. All subjects had been treated for any dental caries or periodontal disease with all restorative treatment completed at least 1 year ago and had ≤3 residual pockets. They were categorized based on a past diagnosis of chronic periodontitis into "healthy" (H) or "periodontitis" (P) categories. Unstimulated whole saliva was collected, genomic DNA was isolated, and high throughput Illumina MiSeq sequencing of 16S rRNA (V3-V4) gene amplicons was performed. The sequences were assigned taxonomy at the species level by using a BLASTN based algorithm that used a combined reference database of HOMD RefSeqV14.51, HOMD RefSeqExtended V1.1 and GreenGeneGold. Species-level OTUs were subjected to downstream analysis in QIIME and R. For P and H group comparisons, community diversity measures were compared, differentially abundant species were determined using DESeq2, and disease indicator species were determined using multi-level pattern analysis within the R package "indicspecies." Results: P subjects were significantly older than H subjects (p = 0.003) but not significantly different in their BOP scores (p = 0.82). No significant differences were noted in alpha diversity measures after adjusting for age, gender, and BOP or in the beta diversity estimates. Four species; Treponema sp. oral taxon 237, TM7 sp. Oral Taxon A56, Prevotella sp. oral taxon 314, Prevotella sp. oral taxon 304, and Capnocytophaga leadbetteri were significantly more abundant in P than in the H group. Indicator species analysis showed 7 significant indicators species of P group. Fusobacterium sp oral taxon 370 was the sole positive indicator of P group (positive predictive value = 0.9, p = 0.04). Significant indicators of the H category were Leptotrichia buccalis, Corynebacterium matruchotii, Leptotrichia hofstadii, and Streptococcus intermedius. Conclusion: This exploratory study showed salivary microbial species could discriminate treated, well-maintained chronic periodontitis from healthy controls with similar gingival inflammation levels. The findings suggest that certain salivary microbiome features may identify periodontitis-susceptible individuals despite clinical disease resolution.


Assuntos
Microbiota , Periodontite/microbiologia , Saliva/microbiologia , Adulto , Idoso , Biodiversidade , Estudos de Casos e Controles , Biologia Computacional , DNA Bacteriano , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Hong Kong/epidemiologia , Humanos , Masculino , Metagenoma , Metagenômica/métodos , Pessoa de Meia-Idade , Periodontite/epidemiologia , Filogenia , RNA Ribossômico 16S/genética
7.
Microbiol Resour Announc ; 8(41)2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31601671

RESUMO

Streptococcus salivarius strain LAB813 was isolated from the dental plaque biofilm of a caries-free child with healthy oral tissues. We report here the complete genome sequence of S. salivarius strain LAB813. This genome consists of a chromosome of 2.2 Mb and a megaplasmid, pSAL813, of 183 kb.

8.
Clin Oral Implants Res ; 30(8): 760-776, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31102416

RESUMO

OBJECTIVE: Periodontitis and peri-implantitis are oral infectious-inflammatory diseases that share similarities in their pathology and etiology. Our objective was to characterize the single-site subgingival and submucosal microbiomes of implant-rehabilitated, partially dentate Chinese subjects (n = 18) presenting with both periodontitis and peri-implantitis. MATERIALS AND METHODS: Subgingival/submucosal plaque samples were collected from four clinically distinct sites in each subject: peri-implantitis submucosa (DI), periodontal pocket (DT), clinically healthy (unaffected) peri-implant submucosa (HI), and clinically healthy (unaffected) subgingival sulcus (HT). The bacterial microbiota present was analyzed using Illumina MiSeq sequencing. RESULTS: Twenty-six phyla and 5,726 operational taxonomic units (OTUs, 97% sequence similarity cutoff) were identified. Firmicutes, Proteobacteria, Fusobacteria, Bacteroidetes, Actinobacteria, Synergistetes, TM7, and Spirochaetes comprised 99.6% of the total reads detected. Bacterial communities within the DI, DT, HI, and HT sites shared high levels of taxonomic similarity. Thirty-one "core species" were present in >90% sites, with Streptococcus infantis/mitis/oralis (HMT-070/HMT-071/HMT-638/HMT-677) and Fusobacterium sp. HMT-203/HMT-698 being particularly prevalent and abundant. Beta-diversity analyses (PERMANOVA test, weighted UniFrac) revealed the largest variance in the microbiota was at the subject level (46%), followed by periodontal health status (4%). Differing sets of OTUs were associated with periodontitis and peri-implantitis sites, respectively. This included putative "periodontopathogens," such as Prevotella, Porphyromonas, Tannerella, Bacteroidetes [G-5], and Treponema spp. Interaction network analysis identified several putative patterns underlying dysbiosis in periodontitis/peri-implantitis sites. CONCLUSIONS: Species (OTU) composition of the periodontal and peri-implant microbiota varied widely between subjects. The inter-subject variations in subgingival/submucosal microbiome composition outweighed differences observed between implant vs. tooth sites, or between diseased vs. healthy (unaffected) peri-implant/periodontal sites.


Assuntos
Placa Dentária , Microbiota , Peri-Implantite , Periodontite , Bactérias , Humanos
9.
Mol Oral Microbiol ; 33(6): 420-429, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30298644

RESUMO

The oral pathogen Streptococcus mutans communicates using a canonical Gram-positive quorum sensing system, CSP-ComDE. The CSP pheromone already known to be involved in the development of genetic competence positively influences the formation of persisters, dormant variants of regular cells that are highly tolerant to antimicrobial therapy. It is now believed that the persistence phenotype is the end result of a stochastic switch in the expression of toxin-antitoxin (TA) modules. TAs consist of a pair of genes that encode two components, a stable toxin and its cognate labile antitoxin. Transcription analyses revealed that three core genes encoding a putative TA system, called SmuATR, were members of the S. mutans CSP regulon. We hypothesized that S. mutans is using its CSP-ComDE system as a deterministic mechanism for persister formation through the activation of smuATR locus. We showed here that the SmuATR system constitutes a novel tripartite type II TA system in which the smuA and smuT genes encode an antitoxin and a toxin, respectively, while SmuR is a transcriptional repressor involved in the autoregulation of the operon. Ectopic expression of SmuA - SmuT is associated with the CSP-inducible persistence phenotype. In contrast, overexpression of SmuT alone is bactericidal and causes membrane permeabilization. To our knowledge, SmuATR is the first functional chromosomal tripartite TA system shown to be induced by the bacterial quorum sensing system and involved in persister formation.


Assuntos
Proteínas de Bactérias/genética , Percepção de Quorum , Streptococcus mutans/genética , Sistemas Toxina-Antitoxina , Proteínas de Bactérias/metabolismo , Regulação Bacteriana da Expressão Gênica , Óperon , Streptococcus mutans/fisiologia , Fatores de Transcrição/genética
10.
Arch Oral Biol ; 83: 169-173, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28780386

RESUMO

OBJECTIVE: The advent of high-throughput sequencing and 'omic' technologies is facilitating an 'open-ended' understanding of the human microbial community and its interplay with health. This commentary aims to present key perspectives and summarize current evidence from metagenomic studies of salivary microbiota in relation to general health and systemic diseases. DESIGN: A narrative review of studies that described salivary microbiome composition in relation to various general health conditions was conducted and the main results were summarized. RESULTS: Currently available evidence shows salivary microbial patterns and fingerprints as related to a range of metabolic, autoimmune and immunodeficiency associated conditions, similar to albeit at a far lower scale than similar studies in the gut microbiome. CONCLUSIONS: Considering the relative ease of collection, emerging evidence of association with non-oral diseases may imply that saliva microbiome research may have potential diagnostic or prognostic value.


Assuntos
Disbiose/microbiologia , Microbiota , Saliva/microbiologia , Dieta , Microbioma Gastrointestinal , Geografia , Humanos , Estilo de Vida
11.
Appl Environ Microbiol ; 83(3)2017 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-27864174

RESUMO

More than 75 "species-level" phylotypes of spirochete bacteria belonging to the genus Treponema reside within the human oral cavity. The majority of these oral treponeme phylotypes correspond to as-yet-uncultivated taxa or strains of uncertain standing in taxonomy. Here, we analyze phylogenetic and taxonomic relationships between oral treponeme strains using a multilocus sequence analysis (MLSA) scheme based on the highly conserved 16S rRNA, pyrH, recA, and flaA genes. We utilized this MLSA scheme to analyze genetic data from a curated collection of oral treponeme strains (n = 71) of diverse geographical origins. This comprises phylogroup 1 (n = 23) and phylogroup 2 (n = 48) treponeme strains, including all relevant American Type Culture Collection reference strains. The taxonomy of all strains was confirmed or inferred via the analysis of ca. 1,450-bp 16S rRNA gene sequences using a combination of bioinformatic and phylogenetic approaches. Taxonomic and phylogenetic relationships between the respective treponeme strains were further investigated by analyzing individual and concatenated flaA (1,074-nucleotide [nt]), recA (1,377-nt), and pyrH (696-nt) gene sequence data sets. Our data confirmed the species differentiation between Treponema denticola (n = 41) and Treponema putidum (n = 7) strains. Notably, our results clearly supported the differentiation of the 23 phylogroup 1 treponeme strains into five distinct "species-level" phylotypes. These respectively corresponded to "Treponema vincentii" (n = 11), Treponema medium (n = 1), "Treponema sinensis" (Treponema sp. IA; n = 4), Treponema sp. IB (n = 3), and Treponema sp. IC (n = 4). In conclusion, our MLSA-based approach can be used to effectively discriminate oral treponeme taxa, confirm taxonomic assignment, and enable the delineation of species boundaries with high confidence. IMPORTANCE: Periodontal diseases are caused by persistent polymicrobial biofilm infections of the gums and underlying tooth-supporting structures and have a complex and variable etiology. Although Treponema denticola is strongly associated with periodontal diseases, the etiological roles of other treponeme species/phylotypes are less well defined. This is due to a paucity of formal species descriptions and a poor understanding of genetic relationships between oral treponeme taxa. Our study directly addresses these issues. It represents one of the most comprehensive analyses of oral treponeme strains performed to date, including isolates from North America, Europe, and Asia. We envisage that our results will greatly facilitate future metagenomic efforts aimed at characterizing the clinical distributions of oral treponeme species/phylotypes, helping investigators to establish a more detailed understanding of their etiological roles in periodontal diseases and other infectious diseases. Our results are also directly relevant to various polymicrobial tissue infections in animals, which also involve treponeme populations.


Assuntos
Proteínas de Bactérias/genética , Variação Genética , Treponema/genética , Flagelina/genética , Tipagem de Sequências Multilocus , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Recombinases Rec A/genética , Análise de Sequência de DNA , Treponema/classificação
12.
Indian J Microbiol ; 56(1): 35-45, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26843695

RESUMO

A culture-independent diversity assessment of archaea, bacteria and fungi in the Thar Desert in India was made. Six locations in Ajmer, Jaisalmer, Jaipur and Jodhupur included semi-arid soils, arid soils, arid sand dunes, plus arid cryptoendolithic substrates. A real-time quantitative PCR approach revealed that bacteria dominated soils and cryptoendoliths, whilst fungi dominated sand dunes. The archaea formed a minor component of all communities. Comparison of rRNA-defined community structure revealed that substrate and climate rather than location were the most parsimonious predictors. Sequence-based identification of 1240 phylotypes revealed that most taxa were common desert microorganisms. Semi-arid soils were dominated by actinobacteria and alpha proteobacteria, arid soils by chloroflexi and alpha proteobacteria, sand dunes by ascomycete fungi and cryptoendoliths by cyanobacteria. Climatic variables that best explained this distribution were mean annual rainfall and maximum annual temperature. Substrate variables that contributed most to observed diversity patterns were conductivity, soluble salts, Ca(2+) and pH. This represents an important addition to the inventory of desert microbiota, novel insight into the abiotic drivers of community assembly, and the first report of biodiversity in a monsoon desert system.

13.
Microb Pathog ; 94: 90-103, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26686411

RESUMO

Bacterial taxa belonging to the phylum Synergistetes are commonly detected within diseased periodontal niches, but are rarely found within healthy oral sites. However, as they typically constitute a minor fraction of the oral microbiota, their precise distributions and disease-associations remain to be fully established. Here, we surveyed the Synergistetes taxa present within individual periodontal/subgingival and peri-implant/submucosal sites, within Chinese subjects (n = 18) affected by both peri-implantitis and periodontitis. Four individual, clinically-distinct sites were analyzed in each patient: healthy sulcus; periodontitis lesion; healthy peri-implant space; peri-implantitis lesion. We employed a clone library-based approach, using PCR-primers that specifically amplified ca. 650bp regions of the 16S rRNA gene from oral cluster A and B Synergistetes taxa. Twenty-one of the 72 sites (from 12/18 subjects) yielded Synergistetes 16S rRNA PCR products. Sequencing of cloned amplicon libraries yielded 1338 quality-filtered 16S rRNA sequences, which were assigned to 26 Synergistetes operational taxonomic units (OTUs; oral taxon SH01-SH26) using a 98.5% identity cut-off. We identified 25 Synergistetes oral cluster A OTUs (genus Fretibacterium; corresponding to Human Oral Taxon (HOT) numbers 358, 359, 360, 361, 362, 363, 452, and 453), and one oral cluster B OTU (Pyramidobacter piscolens oral taxon SH04, HOT-357). Three OTUs predominated: Fretibacterium oral taxon SH01 (HOT-360), Fretibacterium oral taxon SH02 (HOT-452), and Fretibacterium fastidiosum oral taxon SH03 (HOT-363). The Synergistetes community compositions within the respective periodontal and peri-implant sites were variable and complex, and no statistically-significant correlations could be established. However, the detection frequency of F. fastidiosum SH03 and Fretibacterium oral taxon SH01 were both positively associated with plaque index at healthy subgingival sites. Taken together, our results show that diverse Synergistetes populations inhabit both diseased and healthy periodontal and peri-implant niches, with considerable site-to-site variations in composition occurring within the same oral cavity.


Assuntos
Bactérias Anaeróbias Gram-Negativas/isolamento & purificação , Boca/microbiologia , Peri-Implantite/microbiologia , Doenças Periodontais/microbiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biofilmes , China , DNA Bacteriano/genética , Placa Dentária/microbiologia , Feminino , Bactérias Anaeróbias Gram-Negativas/genética , Humanos , Masculino , Microbiota , Pessoa de Meia-Idade , Periodonto/microbiologia , Filogenia , RNA Ribossômico 16S/genética
14.
Microb Pathog ; 94: 76-89, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26550763

RESUMO

This study explored the range of bacterial taxa present within healthy subgingival (below the gum-line) niches in the horse oral cavity using 16S rRNA gene amplicon pyrosequencing. Pooled subgingival plaque samples were collected from approximately 200 sulcus sites from two horses (EQ1, EQ2) for analysis. A total of 14,260 quality-filtered pyrosequencing reads were obtained, which were assigned to 3875 operational taxonomic units (OTUs; 99% identity cut-off); 1907 OTUs for EQ1 and 2156 OTUs for EQ2. Diverse taxa from 12 phyla were identified, including Actinobacteria (3.17%), Bacteroidetes (25.11%), Chloroflexi (0.04%), Firmicutes (27.57%), Fusobacteria (5.15%), Proteobacteria (37.67%), Spirochaetes (0.15%), Synergistetes (0.22%), Tenericutes (0.16%), GN02 (0.19%), SR1 (0.01%) and TM7 (0.37%). Many OTUs were not closely related to known phylotypes, and may represent 'equine-specific' taxa. Phylotypes corresponding to Gammaproteobacteria were abundant, including Actinobacillus spp. (8.75%), unclassified Pasteurellaceae (9.90%) and Moraxella spp. (9.58%). PCR targeting the Synergistetes and Spirochaetes phyla was performed, and resultant plasmid libraries of 16S rRNA gene amplicons (ca. 1480 bp) were Sanger sequenced. Twenty-six Spirochaetes OTUs, and 16 Synergistetes OTUs were identified (99% identity cut-off). These 'species-level' OTUs were assigned Equine Oral Taxon (EOT) numbers, whose phylogenies and taxonomy were comprehensively investigated, in conjunction with corresponding Synergistetes and Spirochaetes OTUs identified by pyrosequencing. The vast majority of Spirochaetes taxa belonged to the genus Treponema, which corresponded to 7 of the 10 human oral treponeme phylogroups. Other Spirochaetes taxa belonging to the Leptospiraceae family were observed; but many treponemes commonly implicated in animal hoof/foot and non-oral soft tissue infections; e.g. Treponema phagedenis, Treponema pedis, Treponema refringens, Treponema calligyrum; were not identified here. Diverse Synergistetes taxa corresponding to oral clusters A and B were identified, which included Fretibacterium fastidiosum and Pyramidobacter piscolens. Taken together, our data reveals that equine subgingival plaque microbiota shares many similarities with the human, canine and feline oral microbiomes.


Assuntos
Bactérias/classificação , Gengiva/microbiologia , Cavalos/microbiologia , Microbiota , Animais , Bactérias/genética , Bactérias/isolamento & purificação , Sequência de Bases , Gatos , DNA Bacteriano/genética , Placa Dentária/microbiologia , Cães , Humanos , Boca/microbiologia , Filogenia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética
15.
Front Microbiol ; 6: 1142, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26539174

RESUMO

Corals are rapidly declining globally due to coral diseases. Skeletal growth anomalies (SGA) or "coral tumors" are a group of coral diseases that affect coral reefs worldwide, including Hong Kong waters in the Indo-Pacific region. To better understand how bacterial communities may vary in corals with SGA, for the first time, we examined the bacterial composition associated with the apparently healthy and the diseased tissues of SGA-affected Platgyra carnosus using 16S ribosomal rRNA gene pyrosequencing. Taxonomic analysis revealed Proteobacteria, Bacteroidetes, Cyanobacteria, and Actinobacteria as the main phyla in both the apparently healthy and the diseased tissues. A significant difference in the bacterial community composition was observed between the two conditions at the OTU level. Diseased tissues were associated with higher abundances of Acidobacteria and Gemmatimonadetes, and a lower abundance of Spirochaetes. Several OTUs belonging to Rhodobacteraceae, Rhizobiales, Gammaproteobacteria, and Cytophaga-Flavobacterium-Bacteroidetes (CFB) were strongly associated with the diseased tissues. These groups of bacteria may contain potential pathogens involved with the development of SGA or opportunistic secondary or tertiary colonizers that proliferated upon the health-compromised coral host. We suggest that these bacterial groups to be further studied based on inoculation experiments and testing of Koch's postulates in efforts to understand the etiology and progression of SGA.

16.
Genome Announc ; 2(6)2014 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-25540346

RESUMO

The oral treponeme bacterium Treponema sp. OMZ 838 was originally isolated from a human necrotizing ulcerative gingivitis (NUG) lesion. Its taxonomic status remains uncertain. The complete genome sequence length was determined to be 2,708,067 bp, with a G+C content of 44.58%, and 2,236 predicted coding DNA sequences (CDS).

17.
Genome Announc ; 2(5)2014 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-25342686

RESUMO

The oral spirochete bacterium Treponema putidum inhabits human periodontal niches. The complete genome sequence of the OMZ 758(T) (ATCC 700334(T)) strain of this species was determined, revealing a 2,796,913-bp chromosome, with a G+C content of 37.30% and a single plasmid (pTPu1; 3,649 bp) identical to pTS1 from Treponema denticola.

18.
Mar Pollut Bull ; 85(2): 400-9, 2014 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-24878302

RESUMO

Hypoxia alters the oxidation-reduction balance and the biogeochemical processes in sediments, but little is known about its impacts on the microbial community that is responsible for such processes. In this study, we investigated the effects of hypoxia and the ubiquitously dispersed flame-retardant BDE47 on the bacterial communities in marine surface sediments during a 28-days microcosm experiment. Both hypoxia and BDE47 alone significantly altered the bacterial community and reduced the species and genetic diversity. UniFrac analysis revealed that BDE47 selected certain bacterial species and resulted in major community shifts, whereas hypoxia changed the relative abundances of taxa, suggesting slower but nonetheless significant community shifts. These two stressors targeted mostly different taxa, but they both favored Bacteroidetes and suppressed Gammaproteobacteria. Importantly, the impacts of BDE47 on bacterial communities were different under hypoxic and normoxic conditions, highlighting the need to consider risk assessments for BDE47 in a broader context of interaction with hypoxia.


Assuntos
Monitoramento Ambiental , Retardadores de Chama/análise , Sedimentos Geológicos/química , Éteres Difenil Halogenados/análise , Anaerobiose , Bactérias/classificação , Bactérias/efeitos dos fármacos , Bactérias/metabolismo , Retardadores de Chama/farmacologia , Sedimentos Geológicos/microbiologia , Éteres Difenil Halogenados/farmacologia , Hong Kong , Oxigênio/análise , Oxigênio/metabolismo , Filogenia
19.
Microb Ecol ; 68(2): 351-9, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24671755

RESUMO

The Antarctic Dry Valleys are unable to support higher plant and animal life and so microbial communities dominate biotic ecosystem processes. Soil communities are well characterized, but rocky surfaces have also emerged as a significant microbial habitat. Here, we identify extensive colonization of weathered granite on a landscape scale by chasmoendolithic microbial communities. A transect across north-facing and south-facing slopes plus valley floor moraines revealed 30-100 % of available substrate was colonized up to an altitude of 800 m. Communities were assessed at a multidomain level and were clearly distinct from those in surrounding soils and other rock-inhabiting cryptoendolithic and hypolithic communities. All colonized rocks were dominated by the cyanobacterial genus Leptolyngbya (Oscillatoriales), with heterotrophic bacteria, archaea, algae, and fungi also identified. Striking patterns in community distribution were evident with regard to microclimate as determined by aspect. Notably, a shift in cyanobacterial assemblages from Chroococcidiopsis-like phylotypes (Pleurocapsales) on colder-drier slopes, to Synechococcus-like phylotypes (Chroococcales) on warmer-wetter slopes. Greater relative abundance of known desiccation-tolerant bacterial taxa occurred on colder-drier slopes. Archaeal phylotypes indicated halotolerant taxa and also taxa possibly derived from nearby volcanic sources. Among the eukaryotes, the lichen photobiont Trebouxia (Chlorophyta) was ubiquitous, but known lichen-forming fungi were not recovered. Instead, fungal assemblages were dominated by ascomycetous yeasts. We conclude that chasmoendoliths likely constitute a significant geobiological phenomenon at lower elevations in granite-dominated Antarctic Dry Valley systems.


Assuntos
Biodiversidade , Ecossistema , Microbiologia do Solo , Regiões Antárticas , Archaea/classificação , Archaea/genética , Bactérias/classificação , Bactérias/genética , Cianobactérias/classificação , Cianobactérias/genética , Fungos/classificação , Fungos/genética , Líquens/classificação , Líquens/genética , Consórcios Microbianos , Microclima , Filogenia , Análise de Sequência de DNA , Dióxido de Silício
20.
Antonie Van Leeuwenhoek ; 104(6): 983-91, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24022396

RESUMO

A novel bacterial strain, JLT2006T, was isolated from the scleractinian coral Platygyra carnosus, located in Hong Kong, China. Cells of this strain were Gram-negative, rod-shaped or oval-shaped and motile by the means of polar flagella. They formed faint-yellow, round colonies on marine agar medium. Phylogenetic analyses based on the 16S rRNA gene sequence indicated that the strain JLT2006T belonged to the class Gammaproteobacteria and was most closely related to Alteromonas-like bacteria of the genera Psychromonas, Pseudoalteromonas, Moritella, Shewanella and Ferrimonas, with less than 93 % sequence similarity. The predominant fatty acids were identified as C18:1x7c/C18:1x6c (23.0 %), C16:1x7c/C16:1x6c (18.2 %) and C16:0 (16.4 %). The quinone was menaquinone-7 (100 %). The polar lipids were determined to be phosphatidylglycerol, phosphatidylethanolamine, phospholipid, glycolipid and lipid. The genomic DNA G?C content was 40.3 mol%. Based on the 16S rRNA gene sequence as well as the physiological and biochemical features that separate the strain JLT2006T from other recognized bacteria, a novel species of a new genus with the name Coralslurrinella hongkonensis gen. nov., sp. nov. is proposed. The type strain is JLT2006T (=JCM 18796T = CGMCC 1.10992T).


Assuntos
Antozoários/microbiologia , Gammaproteobacteria/classificação , Gammaproteobacteria/isolamento & purificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Flagelos/fisiologia , Gammaproteobacteria/genética , Gammaproteobacteria/fisiologia , Hong Kong , Locomoção , Dados de Sequência Molecular , Fosfolipídeos/análise , Filogenia , Quinonas/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
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