BACKGROUND: Colorectal carcinogenesis and progression are related to the gut microbiota and the tumor immune microenvironment. Our previous clinical trial demonstrated that berberine (BBR) hydrochloride might reduce the recurrence and canceration of colorectal adenoma (CRA). The present study aimed to further explore the mechanism of BBR in preventing colorectal cancer (CRC). METHODS: We performed metagenomics sequencing on fecal specimens obtained from the BBR intervention trial, and the differential bacteria before and after medication were validated using quantitative polymerase chain reaction. We further performed ApcMin/+ animal intervention tests, RNA sequencing, flow cytometry, immunohistochemistry, and enzyme-linked immunosorbent assays. RESULTS: The abundance of fecal Veillonella parvula ( V . parvula ) decreased significantly after BBR administration ( P = 0.0016) and increased through the development from CRA to CRC. Patients with CRC with a higher V. parvula abundance had worse tumor staging and a higher lymph node metastasis rate. The intestinal immune pathway of Immunoglobulin A production was activated, and the expression of TNFSF13B (Tumor necrosis factor superfamily 13b, encoding B lymphocyte stimulator [BLyS]), the representative gene of this pathway, and the genes encoding its receptors (interleukin-10 and transforming growth factor beta) were significantly upregulated. Animal experiments revealed that V. parvula promoted colorectal carcinogenesis and increased BLyS levels, while BBR reversed this effect. CONCLUSION: BBR might inhibit V. parvula and further weaken the immunomodulatory effect of B cells induced by V. parvula , thereby blocking the development of colorectal tumors. TRIAL REGISTRAION ClinicalTrials.gov, No. NCT02226185.
Animals ; Humans ; Berberine/therapeutic use* ; Carcinogenesis ; Veillonella ; Colorectal Neoplasms/genetics* ; Tumor Microenvironment

Recurrent aphthous stomatitis (RAS) is a common oral mucosal disorder for which no curative treatment is available. We previously reported that decreased Streptococcus salivarius and increased Acinetobacter johnsonii on the oral mucosa are associated with RAS risk. The purpose of this study was to identify antibiotics that selectively inhibit A. johnsonii but minimally inhibit oral mucosal commensals. S. salivarius KCTC 5512, S. salivarius KCTC 3960, A. johnsonii KCTC 12405, Rothia mucilaginosa KCTC 19862, and Veillonella dispar KCOM 1864 were subjected to antibiotic susceptibility test using amoxicillin, cefotaxime, gentamicin, clindamycin, and metronidazole in liquid culture. The minimal inhibitory concentration (MIC) was defined as the concentration that inhibits 90% of growth. Only gentamicin presented a higher MIC for A. johnsonii than MICs for S. salivarius and several oral mucosal commensals. Interestingly, the growth of S. salivarius increased 10~200% in the presence of sub-MIC concentrations of gentamicin, which was independent of development of resistance to gentamicin. In conclusion, gentamicin may be useful to restore RAS-associated imbalance in oral microbiota by selectively inhibiting the growth of A. johnsonii but enhancing the growth of S. salivarius.
Acinetobacter ; Amoxicillin ; Anti-Bacterial Agents* ; Cefotaxime ; Clindamycin ; Gentamicins ; Mass Screening* ; Metronidazole ; Microbiota ; Mouth Mucosa ; Stomatitis, Aphthous* ; Streptococcus ; Veillonella

The aim of this study was to identify the non-Aggregatibacter actinomycetemcomitans bacteria grown on the tryptic soy-serum-bacitracin-vancomycin (TSBV) medium, an A. actinomycetemcomitans selective medium. A total of 82 unidentified bacterial isolates from the oral cavities of a Korean population were kindly provide by the Korean Collection for Oral Microbiology. All the clinical isolates were grown on TSBV medium and bacterial DNA purified from each isolate was subjected to PCR with universal primers specific for bacterial 16S rRNA genes (16S rDNAs) sequence. The each bacterial 16S rDNA was amplified by PCR and the nucleotide sequences of it was determined by the dideoxynucleotide chain termination method. They were identified by 16S rDNA sequence comparison method at the specie-level. The data showed that Neisseria spp. (42 strains), Fusobacterium spp. (10 strains), Capnocytophaga spp. (8 strains), Propionibacterium acnes (5 strains), Aggregatibacter aprophilus (4 strains), Campylobacter spp. (5 strains), Veillonella dispar (3 strains), Streptococcus sp. (1 strain), Haemophilus parainfluenzae (1 strain), Leptotrichia wadei (1 strain), Morococcus sp./Neisseria sp. (1 strain), and Staphylococcus sp. (1 strain) were identified. These results could be used to develop a new A. actinomycetemcomitans-selective medium which is more effective than the TSBV medium in future studies.
Aggregatibacter ; Bacteria* ; Base Sequence ; Campylobacter ; Capnocytophaga ; DNA, Bacterial ; DNA, Ribosomal ; Fusobacterium ; Genes, rRNA ; Haemophilus parainfluenzae ; Leptotrichia ; Methods ; Neisseria ; Polymerase Chain Reaction ; Propionibacterium acnes ; Staphylococcus ; Streptococcus ; Veillonella

Xylitol is a five-carbon sugar alcohol that reduces the incidence of caries by inhibiting the growth of oral streptococci, including Streptococcus mutans. Since xylitol is transported via the fructose phosphotransferase system, we hypothesized that it could also affect the growth of other oral bacteria strains. We tested the effects of xylitol against non-periodontopathogenic oral bacteria frequently found in healthy subjects as well as periodontopathogens including Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. With 5% xylitol, Streptococcus vestibularis and Gemella morbillorum showed marked growth inhibition. With 10% xylitol, all of the tested periodontopathogens and Actinomyces naeslundii showed marked growth inhibition, whereas the growth inhibition of Neisseria mucosa, Neisseria sicca and Veillonella parvula was mild only. Xylitol is a widely used sweetener and the concentration used in our experiment is easily achieved in the oral cavity. If xylitol reduces the growth of periodontopathogens more preferentially, it could also reduce the prevalence of these pathogens and have clinical utility in the prevention or treatment of periodontal disease.
Actinomyces ; Bacteria* ; Forsythia ; Fructose ; Gemella ; Incidence ; Mouth ; Neisseria mucosa ; Neisseria sicca ; Periodontal Diseases ; Porphyromonas gingivalis ; Prevalence ; Streptococcus ; Streptococcus mutans ; Sweetening Agents ; Treponema denticola ; Veillonella ; Xylitol*

OBJECTIVETo investigate the correlation between the changes in oral microflora and recurrent oral ulcers (ROU).

METHODSSalivary sample were collected from ROU patients with oral ulcers (group T) and those with ulcer healing (group C) as well as from ROU-free individuals (group N). The quantity of 3 common bacteria (Streptococcus sp., Veillonella sp., and Neisseria sp.) in the salivary samples was detected and compared between the 3 groups.

RESULTSThe quantities of Streptococcus sp. (7.30-/+0.89 copies/ml) and Veillonella sp. (8.29-/+0.77 copies/ml) in group T were significantly lower than those in group N (8.15-/+0.55 and 8.93-/+0.76 copies/ml, respectively, P<0.01), but similar with those in group C. The quantity of Streptococcus sp. (7.51-/+0.81 copies/ml) in group C was significantly lower than that in group N (8.15-/+0.55 copies/ml, P<0.01), but the quantity of Veillonella sp. was similar between the two groups. No significant difference were found in the quantity of Neisseria sp. between the 3 groups.

CONCLUSIONThe quantity of oral microflora differs significantly between patients with recurrent oral ulcers and normal individuals, suggesting a possible correlation between oral microfora and recurrent oral ulcers.

Adult ; Colony Count, Microbial ; Female ; Humans ; Male ; Mouth Mucosa ; microbiology ; Neisseria ; isolation & purification ; Saliva ; microbiology ; Stomatitis, Aphthous ; microbiology ; Streptococcus ; isolation & purification ; Veillonella ; isolation & purification

OBJECTIVE: The purpose of this study was to compare the bacterial flora at the peri-implant sulcus of the orthodontic mini-implant placed in the alveolar mucosa with the bacterial flora at the adjacent healthy gingival sulcus. METHODS: Two plaque samples from 7 patients were collected by inserting paper points into the sulcus between the mini-implant and ligature wire connected to the mini-implant head and inflamed alveolar mucosa, and from the gingival sulcus of a healthy tooth adjacent to the mini-implant. RESULTS: Using 16S rDNA clone library, the 24 kinds of bacteria including Haemophilus aphrophilus, Sphingomonas species, Capnocytophaga species, Prevotella melaninogenica, Lachnospiraceae species, Porphyromonas species, Neisseria flava were identified only from the sulcus around the mini-implant. These bacteria constituted only 9.2% of total clones, and the bacteria identified from both the sulcus around mini-implants and the gingival sulcus constituted 80.4% of total clones. Of these bacteria, clones of Prevotella species, Atopobium rimae, Veillonella species, Streptococcus intermedius/constellatus, Streptococcus salivarius were more frequently isolated from the peri-implant sulcus. CONCLUSION: This study suggests that a broad epidemiological study is needed to find causative bacteria which induce inflammation from the peri-implant sulcus.
Aggregatibacter aphrophilus ; Bacteria* ; Capnocytophaga ; Clone Cells* ; DNA, Ribosomal* ; Head ; Humans ; Inflammation ; Ligation ; Mucous Membrane ; Neisseria ; Porphyromonas ; Prevotella ; Prevotella melaninogenica ; Sphingomonas ; Streptococcus ; Tooth ; Veillonella

The purpose of this study was to isolate and identify bacteria from the 4 patients with non-odontogenic infectious lesions (mucormycosis, chronic inflammation from wound infection, and two actinomycosis) and determine their antimicrobial susceptibility against eight antibiotics. Bacterial culture was performed under three culture conditions (anaerobic, CO2, and aerobic incubator). The bacterial strains were identified by 16S rRNA gene (16S rDNA) sequence comparison analysis method. For investigating the antimicrobial susceptibility of the bacteria against eight antibiotics, penicillin G, amoxicillin, tetracycline, cefuroxime, erythromycin, clindamycin, vancomycin, and Augmentin(R) (amoxicillin + clavulanic acid), minimum inhibitory concentration (MIC) measurement was performed using broth microdilution assay. Nosocomial pathogens such as Enterococcus faecalis, Klebsiella pneumoniae, Bacillus subtilis, and Neisseria flavescens were isolated from mucormycosis. Veillonella parvula, Enterobacter hormaechei, and Acinetobacter calcoaceticus were isolated from chronic inflammatory lesion. Actinomyces massiliensis was isolated from actinomycosis in parotid gland. Capnocytophaga ochracea was isolated from actinomycosis in buccal region in anaerobic condition. There was no susceptible antibiotic to all bacteria in mucormycosis. Tetracycline was susceptible to all bacteria in chronic inflammation. C. ochracea was resistant to vancomycin and penicillin G; and other antibiotics showed susceptibility to all bacteria in actinomycosis. The results indicated that the combined treatment of two or more antibiotics is better than single antibiotic treatment in mucormycosis, and penicillin is the first recommended antibiotic to treat actinomycosis.
Acinetobacter calcoaceticus ; Actinomyces ; Actinomycosis ; Amoxicillin ; Anti-Bacterial Agents ; Bacillus subtilis ; Bacteria* ; Capnocytophaga ; Cefuroxime ; Clindamycin ; Enterobacter ; Enterococcus faecalis ; Erythromycin ; Genes, rRNA ; Humans ; Inflammation ; Klebsiella pneumoniae ; Microbial Sensitivity Tests ; Mucormycosis ; Neisseria ; Parotid Gland ; Penicillin G ; Penicillins ; Tetracycline ; Vancomycin ; Veillonella ; Wound Infection

OBJECTIVETo observe the dynamic changes of oral microflora early colonized in infants.

METHODSThe oral swab samples for the study were taken in 1 day, 1, 3, 6, 9, 12 months after birth from 12 healthy neonates. By choosing suitable diluted concentration, the samples were incubated aerobically, facultative anaerobically and anaerobically. The strains were identified by observing colony characteristics, Gram staining and biochemical tests.

RESULTSS. salivarius was the most frequent microflora, followed by S. mitis, S. sanguis, S. gordonii and S. mutans occurred in oral cavity after tooth eruption. Veillonella spp. can be detected in oral cavity of 1-month-old babies, A. odontolyticus was isolated from 8.3% infants of more than 3 months old. L. acidophilus maintained the lower prevalence in oral cavity of babies. Leptotrichia buccalis and Capnocytophaga spp. occurred in oral cavity of some dentate infants.

CONCLUSIONS. solivarius and S. mitis are predominant species in oral cavity of the infants, Veillonella spp. is the first and the most anaerobic species appeared in oral cavity of healthy babies. A. odontolyticus is the first actinomyces detected in oral cavity. With the increasing months, kind and amount of microflora increase dramatically.

Actinomyces ; isolation & purification ; Colony Count, Microbial ; Female ; Humans ; Infant ; Infant, Newborn ; Longitudinal Studies ; Male ; Mouth ; microbiology ; Mouth Mucosa ; microbiology ; Saliva ; microbiology ; Streptococcus ; classification ; isolation & purification ; Streptococcus mitis ; isolation & purification ; Streptococcus mutans ; isolation & purification ; Streptococcus sanguis ; isolation & purification ; Veillonella ; isolation & purification

BACKGROUND: Diverse microbiota exist in the lower respiratory tract. Although next generation sequencing (NGS) is the most widely used microbiome analysis technique, it is difficult to implement NGS in clinical microbiology laboratories. Therefore, we evaluated the performance of conventional culture methods together with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying microbiota in bronchoalveolar lavage (BAL) fluid. METHODS: BAL fluid samples (n=27) were obtained from patients undergoing diagnostic bronchoscopy for lung mass evaluation. Bacterial and fungal culture was performed with conventional media used in clinical microbiology laboratories. On an average, 20 isolated colonies were picked from each agar plate and identified by MALDI-TOF MS. Microbiome analysis using 16S rRNA NGS was conducted for comparison. RESULTS: Streptococcus spp. and Neisseria spp. were most frequently cultured from the BAL fluid samples. In two samples, Enterobacteriaceae grew predominantly on MacConkey agar. Actinomyces and Veillonella spp. were commonly identified anaerobes; gut bacteria, such as Lactobacillus, Bifidobacterium, and Clostridium, and fungi were also isolated. NGS revealed more diverse bacterial communities than culture, and Prevotella spp. were mainly identified solely by NGS. Some bacteria, such as Staphylococcus spp., Clostridium spp., and Bifidobacterium spp., were identified solely by culture, indicating that culture may be more sensitive for detecting certain bacteria. CONCLUSIONS: Culture and NGS of BAL fluid samples revealed common bacteria with some different microbial communities. Despite some limitations, culture combined with MALDI-TOF MS might play a complementary role in microbiome analysis using 16S rRNA NGS.
Actinomyces ; Agar ; Bacteria ; Bifidobacterium ; Bronchoalveolar Lavage Fluid* ; Bronchoalveolar Lavage* ; Bronchoscopy ; Clostridium ; Enterobacteriaceae ; Fungi ; Humans ; Lactobacillus ; Lung ; Mass Spectrometry ; Microbiota ; Neisseria ; Prevotella ; Respiratory System ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Staphylococcus ; Streptococcus ; Veillonella

PURPOSE: Different characteristics of airway microbiome in asthmatics may lead to differential immune responses, which in turn cause eosinophilic or neutrophilic airway inflammation. However, the relationships among these factors have yet to be fully elucidated.METHODS: Microbes in induced sputum samples were subjected to sequence analysis of 16S rRNA. Airway inflammatory phenotypes were defined as neutrophils (>60%) and eosinophils (>3%), and inflammation endotypes were defined by levels of T helper (Th) 1 (interferon-γ), Th2 (interleukin [IL]-5 and IL-13), Th-17 (IL-17), and innate Th2 (IL-25, IL-33, and thymic stromal lymphopoietin) cytokines, inflammasomes (IL-1β), epithelial activation markers (granulocyte-macrophage colony-stimulating factor and IL-8), and Inflammation (IL-6 and tumor necrosis factor-α) cytokines in sputum supernatants was assessed by enzyme-linked immunosorbent assay.RESULTS: The numbers of operational taxonomic units were significantly higher in the mixed (n = 21) and neutrophilic (n = 23) inflammation groups than in the paucigranulocytic inflammation group (n = 19; p < 0.05). At the species level, Granulicatella adiacens, Streptococcus parasanguinis, Streptococcus pneumoniae, Veillonella rogosae, Haemophilus parainfluenzae, and Neisseria perflava levels were significantly higher in the eosinophilic inflammation group (n = 20), whereas JYGU_s levels were significantly higher in the neutrophilic inflammation group compared to the other subtypes (P < 0.05). Additionally, IL-5 and IL-13 concentrations were correlated with the percentage of eosinophils (P < 0.05) and IL-13 levels were positively correlated with the read counts of Porphyromonas pasteri and V. rogosae (P < 0.05). IL-1β concentrations were correlated with the percentage of neutrophils (P < 0.05). had a tendency to be positively correlated with the read count of JYGU_s (P = 0.095), and was negatively correlated with that of S. pneumoniae (P < 0.05).CONCLUSIONS: Difference of microbial patterns in airways may induce distinctive endotypes of asthma, which is responsible for the neutrophilic or eosinophilic inflammation in asthma.
Asthma ; Colony-Stimulating Factors ; Cytokines ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; Haemophilus parainfluenzae ; Inflammasomes ; Inflammation ; Interleukin-13 ; Interleukin-33 ; Interleukin-5 ; Microbiota ; Necrosis ; Neisseria ; Neutrophils ; Phenotype ; Pneumonia ; Porphyromonas ; Sequence Analysis ; Sputum ; Streptococcus ; Streptococcus pneumoniae ; Veillonella