Multispecies Biofilm Development Protocol

Methodology for developing the multispecies biofilm was similar to that previously reported from our research group [14 (link), 23 (link)]. Briefly, reference strains of P. gingivalis ATCC 33277, Streptococcus oralis CECT 907T, Actinomyces naeslundii ATCC 19039, Veillonella parvula NCTC 11810, Fusobacterium nucleatum DMSZ 20482 and Aggregatibacter actinomycetemcomitans DSMZ 8324 were used. Each bacterial strain was grown on blood agar plates (blood agar Oxoid no. 2; Oxoid, Basingstoke, UK), supplemented with 5% (v/v) sterile horse blood (Oxoid), 5.0 mg/L hemin (Sigma, St Louis, MO, USA) and 1.0 mg/L menadione (Merck, Darmstadt, Germany) under anaerobic conditions (10% H₂, 10% CO₂ and 80% N₂) at 37°C for 24–72 h.

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Romero-Lastra P., Sánchez M.C., Llama-Palacios A., Figuero E., Herrera D, & Sanz M. (2019). Gene expression of Porphyromonas gingivalis ATCC 33277 when growing in an in vitro multispecies biofilm. PLoS ONE, 14(8), e0221234.

Publication 2019

blood agar Oxoid no. 2 sterile horse blood hemin menadione

Actinomyces naeslundii Agar Aggregatibacter actinomycetemcomitans Bacterial Biofilm Blood Fusobacterium nucleatum Hemin Horse Menadione Sterile Strain Streptococcus oralis Veillonella parvula

Corresponding Organization : Universidad Complutense de Madrid

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Variable analysis

independent variables

Bacterial strains used: Porphyromonas gingivalis ATCC 33277, Streptococcus oralis CECT 907T, Actinomyces naeslundii ATCC 19039, Veillonella parvula NCTC 11810, Fusobacterium nucleatum DMSZ 20482, and Aggregatibacter actinomycetemcomitans DSMZ 8324

dependent variables

Not explicitly mentioned

control variables

Growth conditions: Blood agar plates supplemented with 5% (v/v) sterile horse blood, 5.0 mg/L hemin, and 1.0 mg/L menadione, under anaerobic conditions (10% H2, 10% CO2 and 80% N2) at 37°C for 24–72 h

controls

No positive or negative controls were explicitly mentioned in the provided information.

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