• Title/Summary/Keyword: Oral microbiology

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Identification of Antibiotic Resistance Genes in Orofacial Abscesses Using a Metagenomics-based Approach: A Pilot Study

  • Yeeun Lee;Joo-Young Park;Youngnim Choi
    • Journal of Korean Dental Science
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    • v.16 no.1
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    • pp.35-46
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    • 2023
  • Purpose: Culture-based methods for microbiological diagnosis and antibiotic susceptibility tests have limitations in the management of orofacial infections. We aimed to profile pus microbiota and identify antibiotic resistance genes (ARGs) using a culture-independent approach. Materials and Methods: Genomic DNA samples extracted from the pus specimens of two patients with orofacial abscesses were subjected to shotgun sequencing on the NovaSeq system. Taxonomic profiling and prediction of ARGs were performed directly from the metagenomic raw reads. Result: Taxonomic profiling revealed obligate anaerobic polymicrobial communities associated with infections of odontogenic origins: the microbial community of Patient 1 consisted of one predominant species (Prevotella oris 74.6%) with 27 minor species, while the sample from Patient 2 contained 3 abundant species (Porphyromonas endodontalis 33.0%; P. oris 31.6%; and Prevotella koreensis 13.4%) with five minor species. A total of 150 and 136 putative ARGs were predicted in the metagenome of each pus sample. The coverage of most predicted ARGs was less than 10%, and only the CfxA2 gene identified in Patient 1 was covered 100%. ARG analysis of the seven assembled genome/metagenome datasets of P. oris revealed that strain C735 carried the CfxA2 gene. Conclusion: A metagenomics-based approach is useful to profile predominantly anaerobic polymicrobial communities but needs further verification for reliable ARG detection.

The influence of Lactobacillus rhamnosus GG on the binding ability of Streptococcus mutans (Lactobacillus rhamnosus GG가 Streptococcus mutans의 부착능에 미치는 영향)

  • Lee, Su-Jun;Lee, Sung-Hoon;Choi, Bong-Kyu;Kim, Jung-Wook;Kim, Young-Jae;Kim, Chong-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.2
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    • pp.155-160
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    • 2011
  • Probiotics has currently attracted for means of preventive treatment measurement instead of using non-specific and broad spectrum antimicrobials. In previous studies, two main probiotics species, Lactobacillus and Bifidobateria, showed the reduction of DMFS and S. mutans counts. However, the timing of introducing probiotic species to oral cavity is not clear. The aim of this study is to evaluate the changes of binding ability of S. mutans in various concentrations and inoculation time of L. rhamnosus GG. Adding the following concentration of L. rhamnosus GG, $1{\times}10^6$ CFU, $1{\times}10^7$ CFU and $1{\times}10^8$ CFU, to S. mutans medium demonstrates significant reduction of S. mutans counts. Additionally, more reduction was observed when L. rhamnosus were inoculated prior to S. mutans or simultaneously inoculated compared to when S. mutans were inoculated prior to L. rhamnosus after 3 hours of incubation. Based on this research, the timing of introducing probiotics should be considered when probiotics are utilized as a preventive treatment measurement.

Alternative Carcinogenicity Screening Assay Using Colon Cancer Stem Cells: A Quantitative PCR (qPCR)-Based Prediction System for Colon Carcinogenesis

  • Bak, Yesol;Jang, Hui-Joo;Shin, Jong-Woon;Kim, Soo-Jin;Chun, Hyun woo;Seo, Ji-Hye;No, Su-Hyun;Chae, Jung-il;Son, Dong Hee;Lee, Seung Yeoun;Hong, Jintae;Yoon, Do-Young
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.645-651
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    • 2018
  • The carcinogenicity of chemicals in the environment is a major concern. Recently, numerous studies have attempted to develop methods for predicting carcinogenicity, including rodent and cell-based approaches. However, rodent carcinogenicity tests for evaluating the carcinogenic potential of a chemical to humans are time-consuming and costly. This study focused on the development of an alternative method for predicting carcinogenicity using quantitative PCR (qPCR) and colon cancer stem cells. A toxicogenomic method, mRNA profiling, is useful for predicting carcinogenicity. Using microarray analysis, we optimized 16 predictive gene sets from five carcinogens (azoxymethane, 3,2'-dimethyl-4-aminobiphenyl, N-ethyl-n-nitrosourea, metronidazole, 4-(n-methyl-n-nitrosamino)-1-(3-pyridyl)-1-butanone) used to treat colon cancer stem cell samples. The 16 genes were evaluated by qPCR using 23 positive and negative carcinogens in colon cancer stem cells. Among them, six genes could differentiate between positive and negative carcinogens with a p-value of ${\leq}0.05$. Our qPCR-based prediction system for colon carcinogenesis using colon cancer stem cells is cost- and time-efficient. Thus, this qPCR-based prediction system is an alternative to in vivo carcinogenicity screening assays.

THE EFFECTS OF SUGARS ON THE EXPRESSION OF GTFB AND GTFC MRNA (Streptococcus mutans의 gtfB 및 gtfC 유전자 발현에 대한 당의 영향)

  • Chung, Hye-Jin;Kim, Shin;Chung, Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.2
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    • pp.299-308
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    • 2007
  • Insoluble glucan is the important component of oral biofilm, which is synthesized from sucrose through the action of glucosyltransferase (GTF) B and GTF C encoded by the gtfB and gtfC genes, respectively of Streptococcus mutans. In present study, the effects of various sugars on the mRNA expression of gtfB and gtfC of S. mutans Ingbritt were examined by fluorescent in situ hybridization (FISH). The mRNA of gtfB and gtfC was expressed normally in the BHI broth containing 1% and 5% sucrose. The mRNA expression was decreased by the addition of 10% of glucose, and 1%, 5% and 10% of fructose. Lactose had no great effect on the expression of gtfB and gtfC. 5% and 10% of xylitol greatly reduced the mRNA expression of gtfB and gtfC. Sorbitol slightly decreased the mRNA expression of gtfB and gtfC when compared to the control. In summary, mRNA expression of gtfB and gtfC was decreased by the addition of glucose, fructose, and xylitol.

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THE EFFECTS OF IONS AND BUFFER SOLUTIONS ON THE MRNA EXPRESSION OF gtfD GENE OF Streptococcus mutans (Streptococcus mutans의 gtfD 유전자 발현에 대한 이온 및 완충액의 영향)

  • Kim, Bo-Young;Kim, Shin;Chung, Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.2
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    • pp.314-322
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    • 2004
  • The production of a glucan was affected by the concentration of ions and buffer solutions, and nutrients in an oral cavity. In this study, the effects of ions and buffer solutions on the mRNA expression of gtfD gene in Streptococcus mutans, an important causative agent of dental caries, were investigated by Fluorescent in situ hybridization(FISH). At first, ions and buffer solutions had little effect on the multiplication of Streptococcus mutans. The green fluorescence according to the mRNA expression of gtfD gene was detected in the BHI broth containing 1% sucrose. The intensities of the green fluorescence were strong at 0.25mM of $CaCl_2$. Little fluorescence was detected by the addition of KCl, except far 10mM KCl at which fluorescence intensities were similar to those of the control. Fluorescence intensities were weak at each concentration of $MgCl_2$ when compared to the control. As for buffer solutions, fluorescence intensities were similar to those of the control at each concentration of buffer solutions, except that they were little detected at 100mM of potassium phosphate.

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Antifungal effect of electrolyzed hydrogen water on Candida albicans biofilm (Candid albicans 바이오필름에 대한 전기분해 수소수의 항진균 효과)

  • Pyo, Kyung-Ryul;Yoo, Yun Seung;Baek, Dong-Heon
    • Journal of Dental Rehabilitation and Applied Science
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    • v.31 no.3
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    • pp.212-220
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    • 2015
  • Purpose: Candida albicans can cause mucosal disease in many vulnerable patients. Also they are associated with denture-related stomatitis. Electrolyzed water is generated by electric current passed via water using various metal electrodes and has antimicrobial activity. The aim of this study was to investigate antifungal activity of electrolyzed water on C. albicans biofilm. Materials and Methods: C. albicans was cultured by sabouraud dextrose broth and F-12 nutrient medium in aerobic and 5% $CO_2$ condition to form blastoconidia (yeast) and hyphae type, respectively. For formation of C. albicans biofilm, C. albicans was cultivated on rough surface 6-well plate by using F-12 nutrient medium in $CO_2$ incubator for 48 hr. After electrolyzing tap water using various metal electrodes, the blastoconidia and hyphal type of C. albicans were treated with electrolyzed water. C. albicans formed blastoconidia and hyphae type when they were cultured by sabouraud dextrose broth and F-12 nutrient medium, respectively. Results: The electrolyzed water using palladium electrode (EWP) exhibited antifungal effect on blastoconidia of C. albicans. Also, the EWP significantly has antifungal activity against C. albicans biofilm and hyphae. In the electrolyzed water using various metal electrodes, only the EWP have antifungal activity. Conclusion: The EWP may use a gargle solution and a soaking solution for prevention of oral candidiasis and denture-related stomatitis due to antifungal activity.

Effect of fluoride-containing gel on the roughness of a titanium surface and the promotion of bacterial growth (불소함유 겔이 티타늄 표면의 세균성 바이오필름 성장에 미치는 영향)

  • Kim, Sun-Jin;Lee, Jae-Kwan;Chang, Beom-Seok;Lee, Si-Young;Um, Heung-Sik
    • Journal of Dental Rehabilitation and Applied Science
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    • v.32 no.1
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    • pp.16-23
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    • 2016
  • Purpose: The aim of this study was to evaluate whether fluorides at various pH cause changes in the surface roughness of titanium implants that alter the adherence of bacterial biofilms. Materials and Methods: The titanium disks were assigned randomly to the following seven groups according to the fluoride agents and application time (1 minute or 30 minute) used: control (no treatment); group 1 (1.23% acidulated phosphate fluoride [APF] at pH 3.5 for 1 minute); group 2 (1.23% APF at pH 3.5 for 30 minute); group 3 (1.23% APF at pH 4.0 for 1 minute); group 4 (1.23% APF at pH 4.0 for 30 minute); group 5 (2% NaF gel at pH 7.0 for 1 minute); group 6 (2% NaF gel at pH 7.0 for 30 minute). The surface roughness of the titanium disks and the amount of adherent bacteria were measured. Results: Group 2 showed a significantly greater surface roughness than the control group (P < 0.0001). No significant differences in the amount of surface bacteria were observed between the treated samples and the controls. In addition, there were no significant differences in bacterial adherence relative to the incubation period between the treated samples and the controls. Conclusion: The surface roughness of the titanium disks was significantly greater after treatment with APF at pH 3.5 for 30 min compared with that of the controls. In addition, we found that the amount of Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibactor actinomycetemcomitans was similar among all groups

Comparison of Cariogenicity of Bovine Milk and Low-fat Milk on Streptococcus mutans Biofilm (국내 시판 우유와 저지방 우유의 Streptococcus mutans 세균막에 대한 우식원성 비교)

  • Hwang, Minseon;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.44 no.2
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    • pp.170-179
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    • 2017
  • The purpose of this study was to compare the cariogenicity of commercially available bovine milk and low-fat milk in a biofilm model using the CDC Biofilm Reactor. Streptococcus mutans ATCC 25175 biofilms were formed on saliva coated bovine enamel slabs in a CDC Biofilm Reactor. Biofilms were exposed three times per day to one of the following materials: commercial whole milk (fat content: 3.4%), low-fat milk (fat content: 1%), or 0.9% NaCl. Medium pH was measured at different time points. After 5 days, biofilms were separated from slabs to evaluate the CFUs. The biofilm thickness was observed by confocal laser-scanning microscopy (CLSM). Enamel slab's demineralization was assessed by measuring surface microhardness before and after the experiment. For microhardness and CFUs assessment, no significant difference was found among the three groups. All groups showed similar pattern of medium pH change and biofilm thickness. Our results showed that there was no difference in the cariogenicity between whole milk and low-fat milk. Both milks were relatively non-cariogenic compared to the control group.

Study of Bacteria Associated with Dental Caries Using a 3 Tone Disclosing Agent (세가지 색상차이를 보이는 착색제를 이용한 치아 우식 관련 균에 관한 연구)

  • Lee, Jeongeun;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.45 no.1
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    • pp.32-40
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    • 2018
  • The aim of this study was to determine the efficacy of a 3 tone plaque disclosing gel in assessing the risk of caries related to the population of Streptococcus mutans, Streptococcus sobrinus, and Lactobacillus spp. quantified using a quantitative real-time polymerase chain reaction (qRT-PCR). 15 healthy children of ages 9 - 12 years were randomly examined. The 3 tone plaque disclosing gel was applied on teeth surfaces, which changed the color to pink or red, blue or purple and light blue. Plaque was divided into 3 groups based on staining. Genomic DNA from each sample was subjected to a qRT-PCR assay for quantitative detection of target bacteria. The Kruskal-Wallis test was conducted for correlation between the color of plaque and the number of bacterial species. The levels of S. mutans, S. sobrinus, and Lactobacillus spp. were significantly different in the plaque samples of the 3 groups (p < 0.05). The proportion of S. sobrinus to S. mutans showed correlation to the color of plaque. The different color-dyed plaque was related to the number of acidogenic bacteria. The 3 tone plaque disclosing gel could be used as one of the indicators to assess the clinical risk of caries associated with the population of S. mutans, S. sobrinus, and Lactobacillus spp.

Surface Roughness and Microbial Adhesion After Finishing of Alkasite Restorative Material (피니싱 처리 이후 알카자이트 수복재의 표면거칠기와 미생물 부착)

  • Park, Choa;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.47 no.2
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    • pp.188-195
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    • 2020
  • This study is aimed to evaluate and compare the surface roughness and microbial adhesion to alkasite restorative material (Cention N), resin-modified glass ionomer (RMGI), and composite resin. And to examine the correlation between bacterial adhesion and surface roughness by different finishing systems. Specimens were fabricated in disk shapes and divided into four groups by finishing methods (control, carbide bur, fine grit diamond bur, and white stone bur). Surface roughness was tested by atomic force microscope and surface observation was performed by scanning electron microscope. Colony forming units were measured after incubating Streptococcus mutans biofilm on specimens using CDC biofilm reactor. Cention N surface roughness was less than 0.2 ㎛ after finishing procedure. Control specimens of resin and Cention N specimens were significantly (p = 0.01) rougher. Pearson correlation coefficient (PCC = 0.13) indicated a weak correlation between surface roughness and S. mutans adhesion to the specimens. Compared with resin specimens, RMGI and Cention N showed lower microbial adhesion. Surface roughness and bacterial adhesion were not significantly different, regardless of the finishing systems.