• Journal of Korean society of Dental Hygiene
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• v.6 no.4
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• pp.311-324
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• 2006

The purpose of this investigation was to evaluate of the specificity of Fusobacterium nucleatum subspecies-specific DNA probes using dot blot hybridization. To confirm whether the clinical isolates were F. nucleatum or not, 16S rDNA of them were cloned and sequenced. The sequencing data were used in homology search with database of GenBank. When the homology was above 98% compared with the nucleotide sequence of a certain bacteria, it was judged as the same species with the bacteria. 23 strains of F. nucleatum were isolates from subgingival plaque of periodontitis patient. The clinical isolates of F. nucleatum were classified into 10 groups using phylogenetic analysis of 16S rDNA sequence. F. nucleatum subspecies nucleatum-specific DNA probe Fu4(1.3 kb) reacted with genomic DNAs from 8 type strains of F. nucleatum and it reacted strongly with those from 8 clinical isolates. The Fp4(0.8 kb) reacted with F. nucleatum subsp. polymorphum ATCC 10953 and one clinical isolates. Fv35(1.9 kb) and Fs17(8.2 kb) probes reacted with genomic DNAs from F. nucleatum subsp. vincentii ATCC 49256 and F. nucleatum subsp. fusiform ATCC 51190, respectively. Our results showed that it is not enough to evaluate the specificity of F. nucleatum subspecies-specific DNA probes with only dot blot hybridization. Therefore, Southern blot analysis will be necessary to confirm the specificity of F. nucleatum subspecies-specific DNA probes.

• Journal of Microbiology
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• v.43 no.4
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• pp.331-336
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• 2005

The objective of this study was to assess the strain-specificity of a DNA probe, Fu12, for Fusobacterium nucleatum subsp. nucleatum ATCC $25586^T$ (F. nucleatum ATCC $25586^T$), and to develop sets of strain-specific polymerase chain reaction (PCR) primers. Strain-specificity was tested against 16 strains of F. nucleatum and 3 strains of distinct Fusobacterium species. Southern blot hybridization revealed that the Fu12 reacted exclusively with the HindIII-digested genomic DNA of F. nucleatum ATCC $25586^T$. The results of PCR revealed that three pairs of PCR primers, based on the nucleotide sequence of Fu12, generated the strain-specific amplicons from F. nucleatum ATCC $25586^T$. These results suggest that the DNA probe Fu12 and the three pairs of PCR primers could be useful in the identification of F. nucleatum ATCC $25586^T$, especially with regard to the determination of the authenticity of the strain.

• Journal of Korean society of Dental Hygiene
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• v.4 no.2
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• pp.153-163
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• 2004

There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from normal inhabitants of children 's oral cavity, which inhibited the the production of halitosis by anaerobic bacteria. The authors identified the isolates by the lest using API 50 CHL medium kit. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30 minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively. 3. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. 4. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 5. When two isolates were tested with API 50 CHL medium kit, those were identified as Lactobaciallius salivarius and Lactobacillus delbrueckii subsp. lactis.

• Journal of dental hygiene science
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• v.10 no.3
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• pp.161-165
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• 2010

The aim of this study was to investigate the antimicrobial effect of (-)-epigalocatechin on Fusobacterium nucleatum, Prevotella intermedia, and Porphyromonas gingivalis. To test the antimicrobial effect of (-)-epigalocatechin, the minimum inhibitory concentration (MIC) of against 4 strains of F. nucleatum, 2 strains of P. intermedia, and 2 strains of P. gingivalis was measured by broth dilution method. Time-kill curves were assessed for susceptible bacteria, testing $0{\times}MIC$ (control group), $0.5{\times}MIC$, $1{\times}MIC$, and $2{\times}MIC$ for (-)-epigalocatechin, by counting viable bacteria after 3, 90, 180, 360, 720, 1440 minutes. The MIC of (-)-epigalocatechin was 0.312-0.625, 0.625, and 0.625 mg/ml on the strains of F. nucleatum, P. intermedia, and P. gingivalis, respectively. Time-kill curves demonstrated (-)-epigalocatechin had bactericidal activity on P. intermedia ATCC $25611^T$, P. gingival is ATCC 53978, and F. nucleatum subsp. fusiforme ATCC $51190^T$. The results suggest that (-)-epigalocatechin can be useful in developing the oral hygiene product such as tooth past and gargling solution for the prevention of periodontal diseases.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.2
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• pp.344-356
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• 2005

There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from inhabitants of caries-free children's oral cavity, which inhibited the formation of artificial plaque by Streptococcus mutans and the production of volatile sulfur compounds by anaerobic bacteria. The isolates were identified by the test using API 50 CHL medium kit and 16S rDNA partial sequencing. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. When Streptococcus mutans was cultured in the media, the mean weight of formed artificial plaque on the orthodontic wires was $124.4{\pm}30.4\;mg$, whereas being reduced to $5.2{\pm}2.0mg$ and $10.6{\pm}6.6mg$ in the media cultured with Streptococcus mutans and each isolate, respectively (p<0.05) 3. The number of viable cells of Streptococcus mutans was $3.4{\times}10^9$ per ml in the cultured solution, whereas those of Streptococcus mutans in the combined culture with each of isolates were $4.6{\times}10^8\;and\;2.4{\times}10^8$ per ml. 4. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively 5. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 6. When two isolates were tested with API 50 CHL medium kit, those were identified Lactobaciallius salivarius and Lactobaciallius delbrueckii subsp. lactis. 7. The similarity values of 16S rDNA sequence between each of isolates and Lactobaciallius salivarius subsp. salicinius were 99.60% and 99.73%, respectively, meaning that isolates were Lactobaciallius salivarius subsp. salicinius. These results indicated that two strains isolated from caries-free children's saliva, which inhibited the formation of artificial plaque and the production of volatile sulfur compounds, were identified as Lactobaciallius salivarius subsp. salicinius.

• Proceedings of the Korean Society of Life Science Conference
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• 2008.10a
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• pp.65.1-65.1
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• 2008

• Genomics & Informatics
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• v.14 no.4
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• pp.255-264
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• 2016

The plethora of genome sequence information of bacteria in recent times has ushered in many novel strategies for antibacterial drug discovery and facilitated medical science to take up the challenge of the increasing resistance of pathogenic bacteria to current antibiotics. In this study, we adopted subtractive genomics approach to analyze the whole genome sequence of the Fusobacterium nucleatum, a human oral pathogen having association with colorectal cancer. Our study divulged 1,499 proteins of F. nucleatum, which have no homolog's in human genome. These proteins were subjected to screening further by using the Database of Essential Genes (DEG) that resulted in the identification of 32 vitally important proteins for the bacterium. Subsequent analysis of the identified pivotal proteins, using the Kyoto Encyclopedia of Genes and Genomes (KEGG) Automated Annotation Server (KAAS) resulted in sorting 3 key enzymes of F. nucleatum that may be good candidates as potential drug targets, since they are unique for the bacterium and absent in humans. In addition, we have demonstrated the three dimensional structure of these three proteins. Finally, determination of ligand binding sites of the 2 key proteins as well as screening for functional inhibitors that best fitted with the ligands sites were conducted to discover effective novel therapeutic compounds against F. nucleatum.

• Korean Journal of Microbiology
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• v.54 no.2
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• pp.146-148
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• 2018

Fusobacterium animalis (formerly Fusobacterium nucleatum subsp. animalis) is a Gram-negative, anaerobic, and filament-shaped bacterium. F. animalis may be a part of normal flora and a periodontopathogen of human oral cavity. F. animalis KCOM 1280 (= ChDC F318) was isolated from a human periodontitis lesion. In this report, we present the draft genome sequence of F. animalis KCOM 1280.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.74-76
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• 2018

Recently, Fusobacterium nucleatum subsp. vincentii was reclassified as Fusobacterium vincentii based on the average nucleotide identity and genome-to-genome distance analyses. F. vincentii is a Gram-negative, anaerobic, and filament-shaped bacterium. F. vincentii is a member of normal flora of human oral cavity and plays a role in periodontal diseases. F. vincentii KCOM 2931 was isolated from a periodontitis lesion. Here, we present the complete genome sequence of F. vincentii KCOM 2931.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.71-73
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• 2018

Recently, Fusobacterium nucleatum subsp. polymorphum was reclassified as Fusobacterium polymorphum based on the average nucleotide identity and genome-to-genome distance analyses. F. polymorphum is a Gram-negative, anaerobic, and filament-shaped bacterium. F. polymorphum is a part of normal flora of oral cavity and causative agent of periodontal diseases. F. polymorphum KCOM 1001 (= ChDC F119) was isolated from a human subgingival plaque of gingivitis lesion. Here, we present the complete genome sequence of F. polymorphum KCOM 1001.