Tear film microbiome in Sjogren’s and non-Sjogren’s aqueous deficiency dry eye

Purpose: To understand the bacterial microbiome changes associated with Sjogren’s syndrome (SS) and non?Sjogren’s syndrome (NSS) aqueous?deficient dry eyes compared to healthy eyes. Methods: Bacterial microbiome was generated from the deoxyribonucleic acid of tear film samples in healthy (n = 33), SS (n = 17), and NSS (n = 28) individuals. Sequencing of the V3?V4 region of the 16S rRNA gene was performed on the Illumina HiSeq2500 platform. Quantitative Insights Into Microbial Ecology (QIIME) pipeline was used to assign taxa to sequences. Statistical analysis was performed in R to assess the alpha diversity and beta diversity indices. Significant changes between the healthy, SS, and NSS cohorts were depicted by principal coordinate analysis (PCoA), differential abundance, and network analysis. Results: Tear microbiome was generated in healthy, SS, and NSS samples. Phyla Actinobacteria, Firmicutes, and Bacteroidetes showed significant changes in SS and NSS compared to healthy. Genera Lactobacillus and Bacillus were predominantly present in all samples. PCoA and heat map analysis showed distinct clusters for SS and NSS from the healthy cohort. Genera Prevotella, Coriobacteriaceae UCG?003, Enterococcus, Streptomyces, Rhodobacter, Ezakiella, and Microbacterium significantly increased in abundance in SS and NSS compared to a healthy cohort. Bacteria–bacteria interaction in SS, NSS, and healthy cohorts was predicted by CoNet network analysis. This analysis predicted a major hub of interaction for the pro?inflammatory bacterium Prevotella in the SS and NSS cohorts. Conclusion: The results of the study indicate significant changes in the phyla and genera in SS and NSS compared to healthy. Both discriminative analysis and network analysis indicated a possible association of predominant pro?inflammatory bacteria with SS and NSS.

Transcriptome analysis of primary human oral keratinocytes stimulated with Prevotella melaninogenica / 口腔疾病防治

Objective@# To investigate the transcriptomic changes in primary human oral keratinocytes (pHOKs) after coculture with Prevotella melaninogenica (P.m) and to verify the changes in human oral keratinocyte (HOK) cell lines.@*Methods@# pHOK was isolated and cocultured with P.m for 0, 4 and 24 h. Total RNA was extracted, a gene library was constructed, transcriptional sequencing was performed, differentially expressed genes (DEGs) were analyzed, gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and the validation of DEGs was performed by qRT-PCR and Western Blot in the HOK and P.m coculture cell model. @*Results @#1 788 DEGs were detected between the 4 h group and control group, including upregulated DEGs such as lymphocyte cytosolic protein 1(LCP1), keratin 7 (KRT7) and Cilia and flagella associated protein 251(CFAP251) and downregulated DEGs such as FERM, ARH/RhoGEF and Pleckstrin domain protein 1 (FARP1), WW domain containing transcription regulator 1(WWTR1) and Discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2). 1 832 DEGs were detected between the 24 h group and control group, including upregulated DEGs such as LCP1, complement C1s(C1S), kynureninase (KYNU) and downregulated DEGs such as phosphoserine aminotransferase 1 (PSAT1), FARP1 and FKBP prolyl isomerase 10 (FKBP10). There were 1 090 common differentially expressed genes (cDEGs) in the 4 h and 24 h groups, including LCP1, KYNU and long intergenic nonprotein coding RNA 958 (LINC00958). The GO pathways were mainly enriched in response to lipopolysaccharide and the molecules of bacterial origin and apical part of the cell. KEGG pathway analysis revealed enrichment in the interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Toll-like receptor (TLR) pathway, etc. We verified the expression of a cDEG, Myosin1B (MYO1B), and qRT-PCR and Western Blot analysis showed that MYO1B expression was significantly upregulated between the control group and the P.m cocultured group (P<0.001), and its expression followed a time-dependent and concentration-dependent manner. @*Conclusion@#P.m played an important role in the transcriptome of oral keratinocytes.

Analysis of microbiota characteristics in tumor tissues sampled under gastroscope / 中华微生物学和免疫学杂志

Objective:To investigate the characteristics of bacterial community in upper gastrointestinal tumors.Methods:The study population was patients with upper gastrointestinal tumors (esophageal cancer and gastric cancer). Gastroscopy was performed on the enrolled patients ( n=17), and the specimens were taken from the tumor sites. At the same time, non-tumor tissues more than 4 cm away from the tumor tissues were taken as the control. After total DNA was extracted and purified, high-throughput 16S DNA gene sequencing was used to detect the microbiota in tumor tissues and control tissues. Bioinformatics analysis was carried out and the differences between groups were compared. Results:16S DNA PCR showed that there was no significant difference in bacterial load between tumor tissues and control tissues. The α-diversity and β-diversity indexes showed that the community composition of the two groups was similar; the samples were discrete and the colony composition was different, but there was no significant difference between the two groups. The results of Venn diagram showed that there were more operational taxonomic units (OTUs) in non-tumor tissues than in tumor tissues (2 068 vs 1 358), indicating that the bacterial species in normal tissues were more abundant than those in tumor tissues. Compared with the control tissues, the percentages of Prevotellaceae ( Prevotella), Lactobacaceae ( Lactobacillus) and Fusobacteriaceae ( Fusobacterium) in tumor tissues were relatively higher (the average percentage was more than twice that of the control). Further paired comparison of the top ten bacteria in the family and genus abundance of the two groups of samples showed that Pseudomonas decreased significantly in tumor tissues at the family ( P=0.041) and genus ( P=0.041) levels, while Prevotella was significantly enriched in tumor tissues at the family ( P=0.031) and genus ( P=0.007) levels. Conclusions:The bacterial community in the tumor microenvironment of the upper gastrointestinal tumor changed, and the species enriched in the tumor site were mainly oral common anaerobic bacteria, such as Prevotellaceae ( Prevotella), Lactobacaceae ( Lactobacillus) and Fusobacteriaceae ( Fusobacterium), especially Prevotellaceae ( Prevotella).

Association between Volatile Sulfur Compounds Prevotella intermedia and Matrix Metalloproteinase-8 Expression

Abstract Objective: To determine the correlation between levels of methyl mercaptan (CH3SH) hydrogen sulfide (H2S), the proportion of Prevotella intermedia (Pi), and matrix metalloproteinase-8 (MMP-8) gene expression levels in periodontitis patients accompanied by halitosis. Material and Methods: Samples were obtained from gingival crevicular fluid (GCF) in the deepest pocket and by swabbing in the tongue coating area in patients with periodontitis presenting with halitosis (n = 23) and healthy subjects as controls (n = 7). The values of CH3SH and H2S were obtained using Oral Chroma. The proportion of Pi and MMP-8 expression levels were evaluated using PCR-RT. All the result was statistically analyzed using SPSS software. Results: The levels of CH3SH and H2S in participants with PD ≥ 6 mm showed a robust negative correlation with the proportion of P. intermedia in GCF and tongue coating. No statistically significant association was detected between CH3SH and H2S levels and MMP-8 expression levels (p>0.05). Conclusion: There is no association between CH3SH and H2S levels, the proportion of P. intermedia, and MMP-8 expression in patients with periodontitis accompanied by halitosis (AU).

Prevalencia de Prevotella spp y Porphyromona spp en periodontitis crónica / Prevalence of Prevotella spp and Porphyromona spp in chronic periodontitis

Las enfermedades del periodonto tienen una etiopatogenia compleja y puede considerarse multifactorial. El factor etiológico esencial en la patología inflamatoria periodontal es la biopelícula dental y cuando el desequilibrio entre el huésped y los microorganismos cambia la complejidad de la flora. Ciertas bacterias como Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens y Treponema spp., han sido comúnmente relacionadas con la periodontitis crónica y son consideradas como indicadores de riesgo para la progresión de dicha enfermedad. El objetivo de este trabajo fue establecer la prevalencia de Prevotella spp y Porphyromona spp en los distintos estadios de periodontitis crónicas. Material y métodos: Se estudiaron 48 pacientes sistémicamente saludables con diagnóstico de periodontitis crónica. Se completó el consentimiento informado, se realizó historia clínica y examen periodontal. El estado periodontal se clasificó en distintos grados de severidad: leve, moderada y severa. Se tomaron muestras de dos sitios con mayor profundidad de sondaje con conos de papel absorbente estériles y se transportaron en un medio prerreducido. Para el aislamiento de Prevotella spp se utilizó agar Brucella más sangre ovina al 5%, hemina, vitamina K al que se agregaron vancomicina y kanamicina; Porphyromonas sp se aisló en el mismo medio con el agregado de bacitracina y colistina. Se sembraron 10 µl de muestra entera y las placas fueron incubadas en jarras de anaerobiosis por 5 a 7 días a 37ºC. Resultados: los distintos grados de periodontitis correspondieron a un 17% periodontits leve, 57% moderada y 26% severa. En el total de pacientes se determinó la presencia de Prevotella spp en el 54% de los casos y un 12,5% de Porphyromona spp. Conclusión: De los pacientes estudiados con periodontits crónica, un 52% correspondió al sexo masculino, un 57% de los casos correspondieron a periodontitis moderada. Se aisló Prevotella sp en todos los estadios de periodontitis crónica y Porphyromonas sp sólo en periodontitis severas (AU)

Periodontal diseases have a complex etiopathogenesis and can be considered multifactorial. The essential etiological factor in periodontal inflammatory pathology is the dental biofilm and when the imbalance between the host and the microorganisms changes the complexity of the flora. Certain bacteria such as Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens and Treponema spp., Have been commonly related to chronic periodontitis and are considered as risk indicators for the progression of said disease. The objective of this work was to establish the prevalence of Prevotella spp and Porphyromonas spp in the different stages of chronic periodontitis. Forty eight systemically healthy patients diagnosed with chronic periodontitis were studied. Informed consent was completed, a medical history and periodontal examination was carried out. The periodontal state was classified into different degrees of severity: mild, moderate and severe. Samples were taken from two sites with greater depth of probing with sterile absorbent paper cones and transported in a prereduced medium. For the isolation of Prevotella spp, Brucella agar plus 5% sheep blood, hemin, vitamin K to which vancomycin and kanamycin were added. For Porphyromonas spp, the same medium was used and bacitracin and colistin were added. 10 µl of the whole sample was seeded and the plates were incubated in anaerobic jars for 5 to 7 days at 37 ° C. Different degrees of periodontitis corresponded to 17% mild periodontitis, 57% moderate and 26% severe. In the total number of patients, the presence of Prevotella spp was determined in 54% of the cases and 12.5% of Porphyromona spp. Of the patients studied with chronic periodontitis, 52% corresponded to the male sex, 57% of the cases corresponded to moderate periodontitis. Prevotella spp was isolated in all stages of chronic periodontitis and Porphyromonas sp only in severe periodontitis (AU)

Isolation and identification of Prevotella nigrescens in patients with chronic periodontitis and analysis of its tumorigenic role in esophageal squamous carcinogenesis / 中华微生物学和免疫学杂志

Objective@#To isolate and identify Prevotella nigrescens (P.nigrescens) in gingival crevicular fluid of patients with chronic periodontitis and to analyze its tumor-promoting role in esophageal squamous cell carcinoma (ESCC).@*Methods@#Samples of gingival crevicular fluid were collected from patients with chronic periodontitis and cultured on GAM agar medium under anaerobic conditions. Black colonies on GAM agar plates were picked for subculture, and then the bacteria were isolated and purified. Bacterial identification was conducted using Gram staining and 16S rDNA sequencing analysis. The isolated bacterial species was used to infect ESCC NE6-T cells and the changes in biological characteristics of the infected cells were evaluated.@*Results@#Multiple bacterial colonies were observed after anaerobic culturing of gingival crevicular fluid samples for 120 h on GAM agar plates. A single bacterial colony with pure black and smooth appearance was obtained from grey black bacterial colonies with streak method. It was a Gram-negative bacterium with bead-like shape. It showed 99.78% 16S rDNA sequence identity with P. nigrescens F0103 and was named P. nigrescens LY01. P. nigrescens LY01 infection promoted the in vitro proliferation, migration and invasion of NE6-T cells and the growth of subcutaneous xenograft in nude mice. In addition, it could also induce the upregulation of Ki67 and activation of p-STAT3.@*Conclusions@#P. nigrescens inhabiting in gingival sulcus might promote the progression of ESCC in human with chronic periodontitis.

Isolation and identification of Prevotella nigrescens in patients with chronic periodontitis and analysis of its tumorigenic role in esophageal squamous carcinogenesis / 中华微生物学和免疫学杂志

Objective To isolate and identify Prevotella nigrescens (P.nigrescens) in gingival cre-vicular fluid of patients with chronic periodontitis and to analyze its tumor-promoting role in esophageal squa-mous cell carcinoma (ESCC). Methods Samples of gingival crevicular fluid were collected from patients with chronic periodontitis and cultured on GAM agar medium under anaerobic conditions. Black colonies on GAM agar plates were picked for subculture, and then the bacteria were isolated and purified. Bacterial identification was conducted using Gram staining and 16S rDNA sequencing analysis. The isolated bacterial species was used to infect ESCC NE6-T cells and the changes in biological characteristics of the infected cells were evaluated. Results Multiple bacterial colonies were observed after anaerobic culturing of gingival cre-vicular fluid samples for 120 h on GAM agar plates. A single bacterial colony with pure black and smooth appearance was obtained from grey black bacterial colonies with streak method. It was a Gram-negative bac-terium with bead-like shape. It showed 99. 78％ 16S rDNA sequence identity with P. nigrescens F0103 and was named P. nigrescens LY01. P. nigrescens LY01 infection promoted the in vitro proliferation, migration and invasion of NE6-T cells and the growth of subcutaneous xenograft in nude mice. In addition, it could al-so induce the upregulation of Ki67 and activation of p-STAT3. Conclusions P.nigrescens inhabiting in gin-gival sulcus might promote the progression of ESCC in human with chronic periodontitis.

The changed abundance of intestinal Prevotella_copri and its predictive value in patients with Parkinson′s disease / 医学研究生学报

ObjectiveGut microbiota plays an important role in Parkinson′s disease, but the mechanisms behind this remain unknown. This study aims to investigate the characteristics of intestinal microbiota in patients with Parkinson′s disease, and to study the changes of intestinal Prevotella_copri and their role in this disease.Methods The study was carried out in 46 patients with Parkinson′s disease and their spouses. The spouse has been living with the patient for a long time, not suffering from any disease. Fecal samples from all subjects were collected using sterile containers. The bacterial DNA was extracted on ice following the Kit protocol. We used the BGISEQ-500 high-throughput sequencing platform to conduct metagenomic shotgun sequencing, to explore the changes of patients′ intestinal microbiota through bioinformatics, and to analyze the function and role of differential microorganisms in disease.ResultsCompared to healthy spouse, the gut microbiota of patients with Parkinson′s disease was significantly changed, which was characterized by decreased Prevotellaceae and Prevotella_copri, but by significantly elevated Bacteroides_stercoris and Escherichia_coli. Prevotella_copri was decreased with age increasing. The correlation analysis showed a significantly negative correlation between the abundance of Prevotella_copri and age, H-Ystage, UPDR total score, and UPDRS Ⅲ score. Results of the random forest model indicated five items including Prevotella_copri had good predictive value for the disease. The functional analysis stated pathways associated with super-pathway of thiamin diphosphate biosynthesis, 4-aminobutanoate degradation. The glucose-1-phosphate degradation and methyl phosphonate degradation significantly increased in patients, while pathways associated with aromatic amino acid biosynthesis, chorismate biosynthesis, thiamin formation, and pyrimidine deoxyribonucleosides salvage significantly decreased. Pathways of Prevotella_copri were mainly concentrated in UMP biosynthesis, S-adenosyl-L-methionine cycle, and guanosine ribonucleotides de novo biosynthesis.ConclusionStructural composition and metabolic functions of gut microbiota were significantly changed in patients with Parkinson′s disease. Prevotella_copri plays an important role in the occurrence and progression of the disease and can be used as a potential biomarker for Parkinson′s disease.

Effect of Bulgarian propolis on the oral microflora in adolescents with plaque-induced gingivitis

ABSTRACT We compared the effect of two therapeutic approaches (marketed toothpaste and addition of Bulgarian propolis extract to the toothpaste) on gingival inflammation, plaque formation and oral microbial flora on Bulgarian adolescents with moderate plaque-induced gingivitis. The participants were divided randomly into two groups of 35 students. The first group was instructed to use marketed toothpaste in their routine oral hygiene. The second group was instructed to add 10 drops of Propolin® to the toothpaste before every brushing. The Gingival index and Plaque index were registered and dental plaque samples were collected on the first visit and on the 20th day of the study. After the treatment, the number of students with Gingival index = 1.1-2.0 in the second group was significantly lower than the respective number in the first group. Neisseria spp. and Streptococcus spp. were present in all samples before and after treatment. The addition of propolis resulted in the complete eradication of Streptococcus mutans, Candida albicans, Fusobacterium varium, Gram-negative cocci, Gram-positive rods, Porphyromonas asaccharolyticus, Prevotella bivia, Prevotella intermedia, Prevotella melani and Streptococcus intermedius. The analyses of Propolin® composition revealed it was a black poplar type propolis and is rich in compounds with pronounced antimicrobial activity. In conclusion, the addition of Bulgarian propolis to the toothpaste improved the gingival health in adolescents with moderate plaque-induced gingivitis and resulted in increased activity against potential periodontal and cariogenic pathogens.

Prevalencia de Prevotella spp y Porphyromona spp en periodontitis crónica / Prevalence of Prevotella spp and Porphyromona spp in chronic periodontitis

Las enfermedades del periodonto tienen una etiopatogenia compleja y puede considerarse multifactorial. El factor etiológico esencial en la patología inflamatoria periodontal es la biopelícula dental y cuando el desequilibrio entre el huésped y los microorganismos cambia la complejidad de la flora. Ciertas bacterias como Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens y Treponema spp., han sido comúnmente relacionadas con la periodontitis crónica y son consideradas como indicadores de riesgo para la progresión de dicha enfermedad. El objetivo de este trabajo fue establecer la prevalencia de Prevotella spp y Porphyromona spp en los distintos estadios de periodontitis crónicas. Material y métodos: Se estudiaron 48 pacientes sistémicamente saludables con diagnóstico de periodontitis crónica. Se completó el consentimiento informado, se realizó historia clínica y examen periodontal. El estado periodontal se clasificó en distintos grados de severidad: leve, moderada y severa. Se tomaron muestras de dos sitios con mayor profundidad de sondaje con conos de papel absorbente estériles y se transportaron en un medio prerreducido. Para el aislamiento de Prevotella spp se utilizó agar Brucella más sangre ovina al 5%, hemina, vitamina K al que se agregaron vancomicina y kanamicina; Porphyromonas sp se aisló en el mismo medio con el agregado de bacitracina y colistina. Se sembraron 10 µl de muestra entera y las placas fueron incubadas en jarras de anaerobiosis por 5 a 7 días a 37ºC. Resultados: los distintos grados de periodontitis correspondieron a un 17% periodontits leve, 57% moderada y 26% severa. En el total de pacientes se determinó la presencia de Prevotella spp en el 54% de los casos y un 12,5% de Porphyromona spp. Conclusión: De los pacientes estudiados con periodontits crónica, un 52% correspondió al sexo masculino, un 57% de los casos correspondieron a periodontitis moderada. Se aisló Prevotella sp en todos los estadios de periodontitis crónica y Porphyromonas sp sólo en periodontitis severas (AU)

Periodontal diseases have a complex etiopathogenesis and can be considered multifactorial. The essential etiological factor in periodontal inflammatory pathology is the dental biofilm and when the imbalance between the host and the microorganisms changes the complexity of the flora. Certain bacteria such as Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens and Treponema spp., Have been commonly related to chronic periodontitis and are considered as risk indicators for the progression of said disease. The objective of this work was to establish the prevalence of Prevotella spp and Porphyromonas spp in the different stages of chronic periodontitis. Forty eight systemically healthy patients diagnosed with chronic periodontitis were studied. Informed consent was completed, a medical history and periodontal examination was carried out. The periodontal state was classified into different degrees of severity: mild, moderate and severe. Samples were taken from two sites with greater depth of probing with sterile absorbent paper cones and transported in a prereduced medium. For the isolation of Prevotella spp, Brucella agar plus 5% sheep blood, hemin, vitamin K to which vancomycin and kanamycin were added. For Porphyromonas spp, the same medium was used and bacitracin and colistin were added. 10 �l of the whole sample was seeded and the plates were incubated in anaerobic jars for 5 to 7 days at 37 ° C. Different degrees of periodontitis corresponded to 17% mild periodontitis, 57% moderate and 26% severe. In the total number of patients, the presence of Prevotella spp was determined in 54% of the cases and 12.5% of Porphyromona spp. Of the patients studied with chronic periodontitis, 52% corresponded to the male sex, 57% of the cases corresponded to moderate periodontitis. Prevotella spp was isolated in all stages of chronic periodontitis and Porphyromonas sp only in severe periodontitis (AU)

The inhibitory effect of ozone water on bad breath pathogens in vitro / 实用口腔医学杂志

Objective: To study the inhibitory effect of ozone water on bad breath pathogens in vitro. Methods: In vitro cultured bad breath pathogens Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi) and Fusobacterium nucleatum (Fn) were identified by Gram stain and a PCR test. Ozonated water by was prepared by ozone generator and the concentration of ozone in water was measured using iodine titration. Artificial saliva was used to observe its interference on ozone function. Results: Gram stain and PCR results were consisted with strain characteristics. The ozone concentration of ozonated water reached to the maximum of 0. 2 mg/L. Ozone water with the concentration was 0. 05, 0. 1 and 0. 2 mg/L inhibited the proliferation of Pg, Pi and Fn. But the inhibitory effect was weakened when the concentration decreased. The artificial saliva reduced the effect of ozone water. Conclusion: The Pg, Pi and Fn can be inhibited by ozone water. Artificial saliva may reduce the effects of ozone water on the bacteria.

The use of Prevotella bryantii 3C5 for modulation of the ruminal environment in an ovine model

Abstract In the Southern Hemisphere, ruminants are mostly raised in grazing systems where animals consume forage and are supplemented with low amounts of concentrates. Concentrates are usually given separately and are rapidly ingested. This practice leads to changing rumen environment conditions during the day, may alter the rumen microbial metabolism and could affect host performance. The native ruminal Prevotella bryantii strain 3C5 was administered every 48 h to wethers under experimental conditions simulating Southern-Hemisphere feeding to evaluate its potential as a rumen fermentation modulator. The inoculum potential was assessed on day 17. The ammonia nitrogen (NH3-N), volatile fatty acids and ruminal pH were monitored on a 24-h basis 19 days after the beginning of the experiment, and the microbial community structure was assessed by pyrosequencing. The administration of P. bryantii modified the fermentation products and daily pH values compared to the control. The NH3-N concentration in the rumen of treated animals was significantly higher than that of the untreated animals. Modification of the ruminal environment and fermentation pathways was achieved without altering the general structure of the microbial community or the potential methane production. P. bryantii 3C5 could be considered in potential probiotic formulations for ruminants in semi-intensive systems.

The use of Prevotella bryantii 3C5 for modulation of the ruminal environment in an ovine model

Abstract In the Southern Hemisphere, ruminants are mostly raised in grazing systems where animals consume forage and are supplemented with low amounts of concentrates. Concentrates are usually given separately and are rapidly ingested. This practice leads to changing rumen environment conditions during the day, may alter the rumen microbial metabolism and could affect host performance. The native ruminal Prevotella bryantii strain 3C5 was administered every 48 h to wethers under experimental conditions simulating Southern-Hemisphere feeding to evaluate its potential as a rumen fermentation modulator. The inoculum potential was assessed on day 17. The ammonia nitrogen (NH3-N), volatile fatty acids and ruminal pH were monitored on a 24-h basis 19 days after the beginning of the experiment, and the microbial community structure was assessed by pyrosequencing. The administration of P. bryantii modified the fermentation products and daily pH values compared to the control. The NH3-N concentration in the rumen of treated animals was significantly higher than that of the untreated animals. Modification of the ruminal environment and fermentation pathways was achieved without altering the general structure of the microbial community or the potential methane production. P. bryantii 3C5 could be considered in potential probiotic formulations for ruminants in semi-intensive systems.

Genome sequence of Prevotella intermedia SUNY aB G8-9K-3, a biofilm forming strain with drug-resistance

Abstract Prevotella intermedia has long been known to be as the principal etiologic agent of periodontal diseases and associated with various systemic diseases. Previous studies showed that the intra-species difference exists in capacity of biofilm formation, antibiotic resistance, and serological reaction among P. intermedia strains. Here we report the genome sequence of P. intermedia SUNY aB G8-9K-3 (designated ATCC49046) that displays a relatively high antimicrobial resistant and biofilm-forming capacity. Genome sequencing information provides important clues in understanding the genetic bases of phenotypic differences among P. intermedia strains.

Red fluorescence of oral bacteria is affected by blood in the growth medium / 대한구강보건학회지

OBJECTIVES: Dental plaque emits red fluorescence under a visible blue light near the ultra-violet end of the light spectrum. The fluorescence characteristics of each microorganism have been reported in several studies. The aim of this study was to evaluate changes in red fluorescence of oral microorganisms that is affected by blood in the culture media. METHODS: The gram-positive Actinomyces naeslundii (AN, KCTC 5525) and Lactobacillus casei (LC, KCTC 3109) and gram negative Prevotella intermedia (PI, KCTC 3692) that are known to emit red fluorescence were used in this study. Each bacterium was activated in broth and cultivated in different agar media at 37℃ for 7 days. Tryptic soy agar with hemin and vitamin K3 (TSA), TSA with sheep blood (TSAB), basal medium mucin (BMM) medium, and BMM with sheep blood (BMMB) were used in this study. Fluorescence due to bacterial growth was observed under 405-nm wavelength blue light using the quantitative light-induced fluorescence-digital (QLF-D) device. The red, green, and blue fluorescence values of colonies were obtained using image-analysis software and the red to green ratio (R/G value) and red to total RGB ratio (R/RGB value) were calculated for quantitative comparison. RESULTS: The QLF-D images of the AN, LC, and PI colonies showed red fluorescence in all media, but the fluorescence of all bacteria was reduced in TSA and BMM media, compared with in TSAB and BMMB media. Both the R/G and the R/RGB values of all bacteria were significantly reduced in growth media without blood (P<0.001). CONCLUSIONS: Based on this in vitro study, it can be concluded that red fluorescence of oral bacteria can be affected by growth components, especially blood. Blood-containing medium could be a significant factor influencing red fluorescence of oral bacteria. It can be further hypothesized that bleeding in the oral cavity can increase the red fluorescence of dental plaque.

Detection of cfxA2, cfxA3, and cfxA6 genes in beta-lactamase producing oral anaerobes

ABSTRACT Purpose The aim of this study was to identify β-lactamase-producing oral anaerobic bacteria and screen them for the presence of cfxA and BlaTEM genes that are responsible for β-lactamase production and resistance to β-lactam antibiotics. Material and Methods Periodontal pocket debris samples were collected from 48 patients with chronic periodontitis and anaerobically cultured on blood agar plates with and without β-lactam antibiotics. Presumptive β-lactamase-producing isolates were evaluated for definite β-lactamase production using the nitrocefin slide method and identified using the API Rapid 32A system. Antimicrobial susceptibility was performed using disc diffusion and microbroth dilution tests as described by CLSI Methods. Isolates were screened for the presence of the β-lactamase-TEM (BlaTEM) and β-lactamase-cfxA genes using Polymerase Chain Reaction (PCR). Amplified PCR products were sequenced and the cfxA gene was characterized using Genbank databases. Results Seventy five percent of patients carried two species of β-lactamase-producing anaerobic bacteria that comprised 9.4% of the total number of cultivable bacteria. Fifty one percent of β-lactamase-producing strains mainly Prevotella, Porphyromonas, and Bacteroides carried the cfxA gene, whereas none of them carried blaTEM. Further characterization of the cfxA gene showed that 76.7% of these strains carried the cfxA2 gene, 14% carried cfxA3, and 9.3% carried cfxA6. The cfxA6 gene was present in three Prevotella spp. and in one Porphyromonas spp. Strains containing cfxA genes (56%) were resistant to the β-lactam antibiotics. Conclusion This study indicates that there is a high prevalence of the cfxA gene in β-lactamase-producing anaerobic oral bacteria, which may lead to drug resistance and treatment failure.

A mouse model of bacterial vaginosis established by infecting estrogen-treated mice with Prevotella bivia / 中华微生物学和免疫学杂志

Objective To establish a mouse model of bacterial vaginosis ( BV) by infecting estro-gen-treated mice with Prevotella bivia ( P. bivia) . Methods The mice were intraperitoneally injected with beta-estradiol 17 valerate which was suspended in sesame oil and then inoculated with different doses of P. bivia strains at the logarithmic phase. Samples of vaginal flushing fluid were collected at different time points after inoculation and used for the isolation of P. bivia strains and the detection of sialidase activities. Altogether 30 mice treated with estrogen and high dose of P. bivia were killed on days 2, 7 and 21 (n=10). Samples of cornua uteri, bladder and kidney were collected from those mice for P. bivia strains isolation. Re-sults Injection the estrogen-treated mice with P. bivia via vagina could cause the P. bivia infection for more than 14 days. The numbers of P. bivia strains isolated on day 21 decreased significantly. Enhanced sialidase activities and clue cells were observed in vaginal secretions of mice with P. bivia infection. Injection of mice with the high dose of P. bivia could spread the infection to cornua uteri. Conclusion Estrogen-treated mice could be used as an animal model for researches on BV.

Analysis of anti-infection treatment for a case of prevotella loescheii infection / 中国药学杂志

OBJECTIVE: To explore the role of clinical pharmacists in drug therapy by assisting the clinician to diagnose and treat a case of disseminated prevotella loescheii infection. METHODS: Clinical pharmacists offered reasonable suggestions for antibiotics selection by analyzing the disease condition of the patient, asking about allergy history, looking up domestic and foreign literature and drug sensitivity test result. And the pharmacists also provided pharmaceutical care and discharging medication education to the patient. RESULTS: The treatment regimen suggested by the clinical pharmacists was adopted by the physicians, and the disease condition was relieved significantly after treatment. CONCLUSION: Clinical pharmacists can demonstrate their value by taking advantage of their pharmacy knowledge and assisting clinicians to develop safe and effective therapeutic regimens.

Presence of Porphyromonas and Prevotella species in the oral microflora of cattle with periodontitis / Presença de espécies de Porphyromonas e Prevotella na microbiota oral de bovinos com periodontite

Bovine periodontitis is a progressive purulent infectious process associated with the presence of strictly and facultative anaerobic subgingival biofilm and epidemiologically related to soil management in large geographic areas of Brazil. This study aimed to detect species of the genera Porphyromonas and Prevotella, which occurr in periodontal pockets of cattle with lesions deeper than 5mm (n=26) and in gingival sulcus of animals considered periodontally healthy (n=25). Presence of the microorganisms was evaluated by independent-culture medium diagnostic method, using polymerase chain reaction (PCR) with specific primers of Porphyromonas asaccharolytica, P. endodontalis, P. gingivalis, P. gulae, Prevotella buccae, P. intermedia, P. loescheii, P. melaninogenica, P. nigrescens, P. oralis and P. tannerae. The species P. endodontalis (80.7%), P. melaninogenica (73.1%) and P. intermedia (61.5%) were the most predominant in samples of cattle with periodontitis. Regarding non-injured gingival sulcus of cattle, P. endodontalis (40%) and P. loeschei (40%) prevailed. Porphyromonas gingivalis, P. gulae and Prevotella tannerae were not detected in the 51 samples studied. Data evaluation by T test, enabled to verify that ocorrence of Porphyromonas asaccharolytica (p=0.000003), P. endodontalis (p=0.0023), Prevotella buccae (p=0.0017), P. intermedia (p=0.0020), P. melaninogenica (p=0.00006) and P. oralis (p=0.0028) is correlated with bovine periodontitis...

A periodontite bovina é um processo infeccioso purulento e progressivo associado à presença de biofilme subgengival anaeróbio estrito e facultativo e de incidência em extensas áreas geográficas do Brasil. O trabalho teve por objetivo detectar espécies dos gêneros Porphyromonas e Prevotella presentes na bolsa periodontal de bovinos com lesões de profundidade maior que 5mm (n=26) e do sulco gengival de animais com idade de 6 a 24 meses e considerados periodontalmente sadios (n=25). A presença dos microrganismos foi avaliada pelo método independente de cultivo bacteriano, por meio da reação em cadeia da polimerase (PCR) com iniciadores específicos para Porphyromonas asaccharolytica, P. endodontalis, P. gingivalis, P. gulae, Prevotella buccae, P. intermedia, P. loescheii, P. melalinogenica, P. nigrescens, P. oralis e P. tannerae. P. endodontalis (80,7%), P. melalinogenica (73,1%) e P. intermedia (61,5%) foram os mais prevalentes nas amostras de bovinos com periodontite. Já no sulco gengival de bovinos sem lesões prevaleceram P. endodontalis (40%) e P. loeschei (40%). Porphyromonas gingivalis, P. gulae e Prevotella tannerae não foram detectados nas 51 amostras pesquisadas. A partir da avaliação dos dados pelo teste T, verificou-se que a ocorrência de Porphyromonas asaccharolytica (p=0.000003), P. endodontalis (p=0.0023), Prevotella buccae (p=0.0017), P. intermedia (p=0.0020), P. melalinogenica (p=0.00006) e P. oralis (p=0.0028) está associada à periodontite bovina...

Prevotella intermedia: uma breve revisão / Prevotella intermedia: A brief review

Prevotella intermedia é um microrganismo anaeróbio Gram negativo, considerado um dos agentes etiológicos de doenças periodontais. Tendo em vista que o biofilme subgengival é um dos seus principais habitats, qualquer quebra na homeostase do meio pode favorecer seu desenvolvimento e consequente dano aos tecidos atingidos. Diversas pesquisas vêm sendo realizadas a fim de identificar o papel deste microrganismo nas doenças periodontais. Assim, este trabalho teve por objetivo revisar a literatura para coletar informações sobre as características e efeitos de Prevotella intermedia no contexto saúde-doença periodontal. A busca foi realizada nas bases de dados PubMed e Lilacs, bem como através de uma busca aleatória. De um total de 47 artigos incluídos, alguns principais resultados puderam ser destacados: Prevotella intermedia está fortemente associada ao quadro de doenças periodontais e suas características favorecem o acometimento e intensificação de doenças diversas. Além disso, este microrganismo é capaz de produzir pigmentos e uma variedade de enzimas as quais favorecem danos aos tecidos periodontais. Existem várias características que aumentam a patogenicidade e virulência deste microrganismo, como por exemplos a sua cápsula polissacarídica, a presença de enzimas protetoras contra o oxigênio, a presença e produção de proteases, além da estimulação de fatores de crescimento no hospedeiro. Não há consenso na literatura acerca da relação de causa-efeito entre presença de Prevotella intermedia e diabetes. Contudo, este microrganismo parece estar envolvido no quadro de pacientes HIV positivo. Por fim, é importante salientar que Prevotella intermedia pode adquirir resistência a alguns tipos de antibióticos, e, por isso, o seu controle é um desafio à clínica atual...

Prevotella intermedia is a Gram negative anaerobic microorganism, considered one of the etiologic agents of periodontal diseases. Taking into account that the subgingival biofilm is its major habitat, any break of the environment homeostasis may lead to the development of the microorganism and consequent damage to the surrounding tissues. Several researches have been conducted in order to identify the role of this microorganism in the periodontal diseases. Thus, this study aimed to review the literature in an attempt to collect information about the characteristics and effects of Prevotella intermedia in the periodontal healthdisease context. The search was performed in the PubMed and Lilacs databases, as well as under a random search. From a total of 47 included articles, some major results could be drawn: Prevotella intermedia is strongly related to the occurrence of periodontal diseases, and its characteristics lead to the incidence of several diseases. Moreover, this microorganism is able to produce pigments and a variety of enzymes, which potentiate the damage of periodontal tissues. There are several characteristics that increase the pathogenicity and virulence of this microorganism, including the polyssacharidic capsule, the presence of protective enzymes against oxygen, the presence and production of proteases, as well as the stimulation of grow factors in the host. There is no consensus in the literature regarding the cause-effect relationship between the presence of Prevotella intermedia and diabetes. However, this microorganism seems to be involved in the occurrence of HIV-positive people. Lastly, it is worth to mention that Prevotella intermedia may acquire strength to some antibiotic types, and, therefore, its control is a challenge to the current clinic...