reference strains s mutans atcc 25175  (ATCC)

Journal: Microorganisms

Article Title: In Vitro Anti-Biofilm and Antibacterial Properties of Streptococcus downii sp. nov.

doi: 10.3390/microorganisms9020450

Figure Lengend Snippet: Primers and probes used for quantification of genomic DNA from the target bacteria. Primers and probes were targeted against 16S rRNA gene (Obtained from Life Technologies Invitrogen (Carlsbad, CA, USA) and Roche (Roche Diagnostic GmbH; Mannheim, Germany)).

Article Snippet: S. downii sp. nov. strain CECT 9732T, isolated from a supragingival dental biofilm sample of an individual with Down syndrome [ ], and the reference bacterial strains S. mutans ATCC 25175, V. parvula NCTC 11810, Actinomyces naeslundii ATCC 19039, F. nucleatum DMSZ 20482, A. actinomycetemcomitans DSMZ 8324, and P. gingivalis ATCC 33277 were used to develop a multi-species biofilm model. Bacteria were cultured anaerobically (10% H 2 , 10% CO 2 , and balance N 2 ) on blood agar plates (Blood Agar Oxoid No 2; Oxoid, Basingstoke, UK), supplemented with 5% ( v/v ) sterile horse blood (Oxoid, Basingstoke, UK), 5.0 mg mL −1 hemin (Sigma-Aldrich, St. Louis, MO, USA) and 1.0 mg mL -1 menadione (Merck, Darmstadt, Germany) for 72 h at 37 °C.

Techniques: Bacteria, Diagnostic Assay, Sequencing

Journal: Microorganisms

Article Title: In Vitro Anti-Biofilm and Antibacterial Properties of Streptococcus downii sp. nov.

doi: 10.3390/microorganisms9020450

Figure Lengend Snippet: Kinetics of incorporation of the six selected bacterial strains in the biofilm [expressed as logarithm of Colony Forming Units per mL, (log CFU mL −1 )] on the two biofilm modalities compared in the study: control biofilms (composed of Streptococcus mutans, Veillonella parvula, Actinomyces naeslundii, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Porphyromonas gingivalis ) and experimental biofilms, incorporating also Streptococcus downii sp. nov. Analyses have been performed with quantitative polymerase chain reaction, in biofilms from 12 h to 120 h of incubation, using specific primers and probes directed to the 16S rRNA gene. * p < 0.005.

Article Snippet: S. downii sp. nov. strain CECT 9732T, isolated from a supragingival dental biofilm sample of an individual with Down syndrome [ ], and the reference bacterial strains S. mutans ATCC 25175, V. parvula NCTC 11810, Actinomyces naeslundii ATCC 19039, F. nucleatum DMSZ 20482, A. actinomycetemcomitans DSMZ 8324, and P. gingivalis ATCC 33277 were used to develop a multi-species biofilm model. Bacteria were cultured anaerobically (10% H 2 , 10% CO 2 , and balance N 2 ) on blood agar plates (Blood Agar Oxoid No 2; Oxoid, Basingstoke, UK), supplemented with 5% ( v/v ) sterile horse blood (Oxoid, Basingstoke, UK), 5.0 mg mL −1 hemin (Sigma-Aldrich, St. Louis, MO, USA) and 1.0 mg mL -1 menadione (Merck, Darmstadt, Germany) for 72 h at 37 °C.

Techniques: Control, Real-time Polymerase Chain Reaction, Incubation

Journal: Microorganisms

Article Title: In Vitro Anti-Biofilm and Antibacterial Properties of Streptococcus downii sp. nov.

doi: 10.3390/microorganisms9020450

Figure Lengend Snippet: Confocal micrographs that represented a 2D maximum projection of the series along fixed axis of the control biofilms ( a , c , e ), composed by Streptococcus mutans, Veillonella parvula, Actinomyces naeslundii, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, and Porphyromonas gingivalis , and experimental biofilms, incorporating also Streptococcus downii sp. nov. ( b , d , f ), after 24 h ( a , b ), 72 h ( c , d ) and 120 h ( e , f ) of growth. LIVE/DEAD ® BacLight TM Bacterial Viability Kit stain was used to assess the vitality of cells (live cells in green and dead cells in red color; yellowish corresponded to damage cells but still alive).

Article Snippet: S. downii sp. nov. strain CECT 9732T, isolated from a supragingival dental biofilm sample of an individual with Down syndrome [ ], and the reference bacterial strains S. mutans ATCC 25175, V. parvula NCTC 11810, Actinomyces naeslundii ATCC 19039, F. nucleatum DMSZ 20482, A. actinomycetemcomitans DSMZ 8324, and P. gingivalis ATCC 33277 were used to develop a multi-species biofilm model. Bacteria were cultured anaerobically (10% H 2 , 10% CO 2 , and balance N 2 ) on blood agar plates (Blood Agar Oxoid No 2; Oxoid, Basingstoke, UK), supplemented with 5% ( v/v ) sterile horse blood (Oxoid, Basingstoke, UK), 5.0 mg mL −1 hemin (Sigma-Aldrich, St. Louis, MO, USA) and 1.0 mg mL -1 menadione (Merck, Darmstadt, Germany) for 72 h at 37 °C.

Techniques: Control, Staining

Journal: Microorganisms

Article Title: In Vitro Anti-Biofilm and Antibacterial Properties of Streptococcus downii sp. nov.

doi: 10.3390/microorganisms9020450

Figure Lengend Snippet: Scanning electron microscope images of the control biofilms ( a , c , e ), composed by Streptococcus mutans, Veillonella parvula, Actinomyces naeslundii, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis , and experimental biofilms, incorporating also Streptococcus downii sp. nov. ( b , d , f ), after 24 h ( a , b ), 72 h ( c , d ) and 120 h ( e , f ) of growth. A similar architecture of biofilms can be observed, in both presence and absence of S. downii sp. nov., with biofilms covering the disc surfaces with flat homogenous layers of cells, combined with bacterial clusters, showing channels inside the structure.

Article Snippet: S. downii sp. nov. strain CECT 9732T, isolated from a supragingival dental biofilm sample of an individual with Down syndrome [ ], and the reference bacterial strains S. mutans ATCC 25175, V. parvula NCTC 11810, Actinomyces naeslundii ATCC 19039, F. nucleatum DMSZ 20482, A. actinomycetemcomitans DSMZ 8324, and P. gingivalis ATCC 33277 were used to develop a multi-species biofilm model. Bacteria were cultured anaerobically (10% H 2 , 10% CO 2 , and balance N 2 ) on blood agar plates (Blood Agar Oxoid No 2; Oxoid, Basingstoke, UK), supplemented with 5% ( v/v ) sterile horse blood (Oxoid, Basingstoke, UK), 5.0 mg mL −1 hemin (Sigma-Aldrich, St. Louis, MO, USA) and 1.0 mg mL -1 menadione (Merck, Darmstadt, Germany) for 72 h at 37 °C.

Techniques: Microscopy, Control

Journal: Microorganisms

Article Title: In Vitro Anti-Biofilm and Antibacterial Properties of Streptococcus downii sp. nov.

doi: 10.3390/microorganisms9020450

Figure Lengend Snippet: Analysis of 24-h biofilms by confocal laser scanning microscopy, showing a typical structure of biofilms, with bacteria in discontinuous microcolonies over hydroxyapatite surfaces, with a majority of live cells (live cells in green; dead cells in red; yellowish corresponded to damage cells but still alive); and counts (expressed as logarithm of colony forming units per mL, CFU mL −1 ) of Streptococcus mutans ( Sm ), Veillonella parvula ( Vp ), Actinomyces naeslundii ( An ), Fusobacterium nucleatum ( Fn ), Aggregatibacter actinomycetemcomitans ( Aa ), and Porphyromonas gingivalis ( Pg ).

Article Snippet: S. downii sp. nov. strain CECT 9732T, isolated from a supragingival dental biofilm sample of an individual with Down syndrome [ ], and the reference bacterial strains S. mutans ATCC 25175, V. parvula NCTC 11810, Actinomyces naeslundii ATCC 19039, F. nucleatum DMSZ 20482, A. actinomycetemcomitans DSMZ 8324, and P. gingivalis ATCC 33277 were used to develop a multi-species biofilm model. Bacteria were cultured anaerobically (10% H 2 , 10% CO 2 , and balance N 2 ) on blood agar plates (Blood Agar Oxoid No 2; Oxoid, Basingstoke, UK), supplemented with 5% ( v/v ) sterile horse blood (Oxoid, Basingstoke, UK), 5.0 mg mL −1 hemin (Sigma-Aldrich, St. Louis, MO, USA) and 1.0 mg mL -1 menadione (Merck, Darmstadt, Germany) for 72 h at 37 °C.

Techniques: Confocal Laser Scanning Microscopy, Bacteria

Journal: Microorganisms

Article Title: In Vitro Anti-Biofilm and Antibacterial Properties of Streptococcus downii sp. nov.

doi: 10.3390/microorganisms9020450

Figure Lengend Snippet: Counts (expressed as logarithm of colony forming units per mL, CFU/mL) of Streptococcus mutans (Sm), Veillonella parvula (Vp), Actinomyces naeslundii (An), Fusobacterium nucleatum (Fn), Aggregatibacter actinomycetemcomitans (Aa) , and Porphyromonas gingivalis (Pg) in biofilms using specific primers and probes directed to the 16S rRNA gene: ( a ) 24-h biofilms after contact with 10 8 CFU/mL of S. downii sp. nov. for 24 h more; ( b ) 24-h biofilms after contact with 10 8 CFU/mL of S. downii sp. nov. for 48 h more. * p < 0.005.

Article Snippet: S. downii sp. nov. strain CECT 9732T, isolated from a supragingival dental biofilm sample of an individual with Down syndrome [ ], and the reference bacterial strains S. mutans ATCC 25175, V. parvula NCTC 11810, Actinomyces naeslundii ATCC 19039, F. nucleatum DMSZ 20482, A. actinomycetemcomitans DSMZ 8324, and P. gingivalis ATCC 33277 were used to develop a multi-species biofilm model. Bacteria were cultured anaerobically (10% H 2 , 10% CO 2 , and balance N 2 ) on blood agar plates (Blood Agar Oxoid No 2; Oxoid, Basingstoke, UK), supplemented with 5% ( v/v ) sterile horse blood (Oxoid, Basingstoke, UK), 5.0 mg mL −1 hemin (Sigma-Aldrich, St. Louis, MO, USA) and 1.0 mg mL -1 menadione (Merck, Darmstadt, Germany) for 72 h at 37 °C.

Techniques:

Journal: Clinical Oral Investigations

Article Title: NF-kB mediated down-regulation of collagen synthesis upon HEMA (2-hydroxyethyl methacrylate) treatment of primary human gingival fibroblast/ Streptococcus mutans co-cultured cells

doi: 10.1007/s00784-014-1304-4

Figure Lengend Snippet: a Light phase contrast microscopy of co-cultured human gingival fibroblasts (HGF)/ S. mutans ATCC 25175 in different experimental conditions. Arrows indicate dead cells. Magnification ×10. b Trypan blue dye exclusion test of primary cultures of HGF exposed for 24 h to 3 mM HEMA co-cultured or not in the presence of S. mutans ATCC 25175. Data are the mean of three separate experiments (±SD). c Effect of 3 mM HEMA on the viable growth of S. mutans ATCC 25175 in co-culture with human gingival fibroblast after 24 h of incubation. Viable growth of S. mutans ATCC 25175 adherent on HGF with and without HEMA. Data are the mean of three separate experiments (±SD). d LDH release in HGF in different experimental conditions. Graph represents the mean percentage ± SD of three experiments. C HGF (control), H HGF + HEMA, M HGF + S. mutans ATCC25175, HM HGF + HEMA + S. mutans ATCC 25175

Article Snippet: The reference strain S. mutans ATCC 25175 has been cultured in Trypticase soy broth (Oxoid, Milan, Italy) at 37 °C for 18–24 h under anaerobic atmosphere.

Techniques: Microscopy, Cell Culture, Co-Culture Assay, Incubation, Control

Journal: Clinical Oral Investigations

Article Title: NF-kB mediated down-regulation of collagen synthesis upon HEMA (2-hydroxyethyl methacrylate) treatment of primary human gingival fibroblast/ Streptococcus mutans co-cultured cells

doi: 10.1007/s00784-014-1304-4

Figure Lengend Snippet: a Representative image of the expression of CoL1A1 normalized with the expression of the internal control GAPDH. b Densitometric analysis (±SD) of CoL1A1 gene expression. c Western blot analysis of procollagen I expression in HGF exposed for 24 h to 3 mM HEMA co-cultured or not in the presence of S. mutans ATCC 25175. Total protein (60 μg) has been loaded for each lane and the membrane probed with β-actin antibody to verify loading evenness. The blot is the most representative out of three independent experiments performed in duplicate. d Densitometric analysis (±SD) of Procollagen I protein expression. For legend, see Fig. ; H , HM versus C pro-collagen I: p < 0.05

Article Snippet: The reference strain S. mutans ATCC 25175 has been cultured in Trypticase soy broth (Oxoid, Milan, Italy) at 37 °C for 18–24 h under anaerobic atmosphere.

Techniques: Expressing, Control, Gene Expression, Western Blot, Cell Culture, Membrane

Journal: Clinical Oral Investigations

Article Title: NF-kB mediated down-regulation of collagen synthesis upon HEMA (2-hydroxyethyl methacrylate) treatment of primary human gingival fibroblast/ Streptococcus mutans co-cultured cells

doi: 10.1007/s00784-014-1304-4

Figure Lengend Snippet: a Western blot analysis of NF-kB, IkBα and p-IkBα expression in HGF exposed for 24 h to 3 mM HEMA co-cultured or not in the presence of S. mutans . When required, the NF-kB/Ikb pharmacological inhibitor SN50 has been added to the culture 45 min before S. mutans and HEMA treatment. Total protein (60 μg) has been loaded for each lane and the membrane probed with β-actin antibody to verify loading evenness. The blot is the most representative out of three independent experiments performed in duplicate. b Densitometric analysis (±SD) of p-IkBα/IkBα ratio. For legend, see Fig. ; H , M , HM versus C p-IkBα: p < 0.05

Article Snippet: The reference strain S. mutans ATCC 25175 has been cultured in Trypticase soy broth (Oxoid, Milan, Italy) at 37 °C for 18–24 h under anaerobic atmosphere.

Techniques: Western Blot, Expressing, Cell Culture, Membrane

Journal: Clinical Oral Investigations

Article Title: NF-kB mediated down-regulation of collagen synthesis upon HEMA (2-hydroxyethyl methacrylate) treatment of primary human gingival fibroblast/ Streptococcus mutans co-cultured cells

doi: 10.1007/s00784-014-1304-4

Figure Lengend Snippet: a Western blot analysis of Bax and Bcl-2 expression in HGF exposed for 24 h to 3 mM HEMA co-cultured or not in the presence of S. mutans. When required, the NF-kB/IkB pharmacological inhibitor SN50 has been added to the culture 45 min before S. mutans and HEMA treatment. Total protein (60 μg) has been loaded for each lane and the membrane probed with β-actin antibody to verify loading evenness. The blot is the most representative out of three independent experiments performed in duplicate. b Densitometric analysis (±SD) of Bax and Bcl-2 expression. For legend, see Fig. ; H , M , HM versus C Bax: p < 0.05; H , M , HM versus C Bcl-2: p < 0.05

Article Snippet: The reference strain S. mutans ATCC 25175 has been cultured in Trypticase soy broth (Oxoid, Milan, Italy) at 37 °C for 18–24 h under anaerobic atmosphere.

Techniques: Western Blot, Expressing, Cell Culture, Membrane

Journal: Clinical Oral Investigations

Article Title: NF-kB mediated down-regulation of collagen synthesis upon HEMA (2-hydroxyethyl methacrylate) treatment of primary human gingival fibroblast/ Streptococcus mutans co-cultured cells

doi: 10.1007/s00784-014-1304-4

Figure Lengend Snippet: ELISA assay for type I collagen secretion of primary cultures of HGF exposed for 24 h to 3 mM HEMA co-cultured or not in the presence of S.mutans . When required, the NF-kB/IkB pharmacological inhibitor SN50 has been added to the culture 45 min before S. mutans and HEMA treatment. Secretion levels have been measured at 24 h and reported as micrograms per millilitre. The results are the mean (±SD) of three different experiments. H-SN50 versus C collagen secretion, p < 0.05; H + SN50 versus C collagen secretion, p < 0.05

Article Snippet: The reference strain S. mutans ATCC 25175 has been cultured in Trypticase soy broth (Oxoid, Milan, Italy) at 37 °C for 18–24 h under anaerobic atmosphere.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture