type strain f nucleatum atcc 23726  (ATCC)

Journal: Journal of Oral Microbiology

Article Title: Antagonistic interaction between two key endodontic pathogens Enterococcus faecalis and Fusobacterium nucleatum

doi: 10.1080/20002297.2022.2149448

Figure Lengend Snippet: The qualitative and quantitative coaggregation assay for E. faecalis ( Ef ) and F. nucleatum ( Fn ) . (a) Coaggregation assay to screen eight different F. nucleatum ATCC23726 outer-membrane protein mutants for defective coaggregation phenotype with E. faecalis OG1RF. The arrow points to the fap2 mutant with binding deficiency. (b) Coaggregation assay to screen sugars and amino acids for their inhibitory efforts on interaction between E. faecalis and F. nucleatum . The arrow points to galactose with inhibitory effort. (c) Microscopic observation of coaggregation between E. faecalis and F. nucleatum under different conditions: a. E. faecalis alone; b. F. nucleatum alone; c. E. faecalis and F. nucleatum ; d. E. faecalis and F. nucleatum fap2 . For each interacting pair, the pictures of 10 random views were taken and 1 representative image was shown. 100X magnification. The scale bar is 10 µm. (d) Spectrophotometric coaggregation assay to quantitatively measure coaggregation between wild type E. faecalis ( Ef WT)/ E. faecalis clinical isolate ( Ef DX1) and wild type F. nucleatum ( Fn WT) and its outer-membrane protein mutants. (e) Spectrophotometric coaggregation assay to quantitatively measure coaggregation between E. faecalis and F. nucleatum in the presence of selected sugars and amino acids, which are D-galactose (Gal), D-Mannose (Man), L-arginine (Arg), L-leucine (Leu) and L-glutamic acid (Glu). All co-aggregation assays have 120 min incubation time. The data presented are based on experiments conducted in triplicate.

Article Snippet: F. nucleatum wild type strain ATCC 23726 and its mutant derivatives defective in outer membrane autotransporter proteins, including Fn 1449( fap2 ), Fn 1526( radD ), Fn 2058( aim1 ), Fn 2047, Fn 0254, Fn 1554, Fn 1253( aid1 ), and Fn 1893 [ ], were maintained under anaerobic conditions (10% H 2 , 10% CO 2 , 80% N 2 ) at 37°C on either Columbia agar supplemented with 5% sheep blood or in Columbia Broth (CB; BD Difco, Detroit, MI, USA).

Techniques: Mutagenesis, Binding Assay, Incubation

Journal: Journal of Oral Microbiology

Article Title: Antagonistic interaction between two key endodontic pathogens Enterococcus faecalis and Fusobacterium nucleatum

doi: 10.1080/20002297.2022.2149448

Figure Lengend Snippet: The acidic environment generated by E. faecalis inhibits F. nucleatum growth . (a) pH challenging assay. F. nucleatum ATCC 23726 viability was tested under different pH conditions in Columbia broth. The data presented are based on experiments conducted in triplicates. WT, wild type. (b) Competition assay under buffered pH condition. Quantitative analysis of the competition effects between E. faecalis ( Ef ) and F. nucleatum ( Fn ) under buffered pH condition. Data are expressed as relative CFU (%) compared with the Fn alone CFU at each timepoint as 100%. Graph shows means and SD of readings from two individual experiments performed in triplicate. Data represent the means and standard deviation of at least three independent experiments. *P < 0.05; ** P < 0.01; *** P < 0.001. WT, wild type.

Article Snippet: F. nucleatum wild type strain ATCC 23726 and its mutant derivatives defective in outer membrane autotransporter proteins, including Fn 1449( fap2 ), Fn 1526( radD ), Fn 2058( aim1 ), Fn 2047, Fn 0254, Fn 1554, Fn 1253( aid1 ), and Fn 1893 [ ], were maintained under anaerobic conditions (10% H 2 , 10% CO 2 , 80% N 2 ) at 37°C on either Columbia agar supplemented with 5% sheep blood or in Columbia Broth (CB; BD Difco, Detroit, MI, USA).

Techniques: Generated, Competitive Binding Assay, Standard Deviation