The fifteen multi-species biofilm that were used to contaminate the titanium discs are the essential bacterial strains found in peri-implantitis sites. The included bacterial strains were Porphyromonas gingivalis (ATCC 33277), Aggregatibacter actinomycetemcomitans (ATCC43718), Prevotella intermedia (ATCC 25611), Fusobacterium nucleatum (ATCC10953), Streptococcus mutans (ATCC 25175), and Streptococcus sobrinus (ATCC 33478). Commensal bacteria were also included in the study in order to mimic as possible the peri-implant biofilm. The commensal bacteria were Actinomyces viscosus (ATCC 15987), Streptococcus cristatus (ATCC 49999), Actinomyces naeslundii (ATCC 51655), Streptococcus mitis (ATCC 49456), Streptococcus oralis (DSM 20627), Streptococcus sanguinis (LMG 14657), Streptococcus gordonii (ATCC 49818), Veillonella parvula (DSM 2008), and Streptococcus parasanguinis (DSM 6778).
Streptococcus mitis
Manufactured by American Type Culture Collection
Sourced in United States
Streptococcus mitis is a bacterial strain from the American Type Culture Collection (ATCC) product catalog. It is a gram-positive, facultative anaerobic coccus that is a normal inhabitant of the human oral cavity. The strain is available for use in microbiological research and applications.
Automatically generated - may contain errors
Lab products found in correlation
Streptococcus mutans, by American Type Culture Collection (3 mentions)
Streptococcus sobrinus, by American Type Culture Collection (3 mentions)
Porphyromonas gingivalis, by American Type Culture Collection (2 mentions)
Streptococcus sanguinis, by American Type Culture Collection (2 mentions)
Streptococcus gordonii, by American Type Culture Collection (1 mentions)
Actinomyces viscosus, by American Type Culture Collection (1 mentions)
Veillonella parvula, by American Type Culture Collection (1 mentions)
Bacteroides fragilis, by American Type Culture Collection (1 mentions)
Prevotella nigrescens, by American Type Culture Collection (1 mentions)
Actinomyces naeslundii, by American Type Culture Collection (1 mentions)
Fusobacterium nucleatum, by American Type Culture Collection (1 mentions)
Bhi broth, by BD (1 mentions)
Yt broth, by BD (1 mentions)
Chlorhexidine digluconate, by Merck Group (1 mentions)
Resazurin, by Merck Group (1 mentions)
Streptococcus salivarius, by American Type Culture Collection (1 mentions)
6 protocols using streptococcus mitis
1
Multi-species Biofilm Peri-implantitis Model
2
Cultivation of Oral Bacterial Species
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Porphyromonas gingivalis ATCC 33277, Veillonella parvula ATCC 17745, Streptococcus mitis ATCC 49456, Neisseria flavescens ATCC 13120, Hemophilus parainfluenzae NCTC 10665, Rothia mucilaginosa ATCC 49041, and Lautropia mirabilis ATCC 51599 were obtained from the American Tissue Culture Collection (ATCC) or Public Health England. V. parvula was cultivated in Brain Heart Infusion (BHI) supplemented with 1.5 of 60% sodium lactate. All other bacteria were cultivated in BHI supplemented with 0.5% hemin, 0.1% Vitamin K and 1% Isovitalex. P. gingivalis and V. parvula were grown in anaerobic conditions (10% hydrogen, 10% CO2, and 80% nitrogen), L. mirabilis was grown aerobically, and the other five bacteria were grown in 5% CO2. All strains were grown at 37°C.
3
Antibacterial Evaluation of Essential Oils
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The tested strains were obtained from the American Type Culture Collection (ATCC, Rockville MD, USA). The following microorganisms were used in the evaluation of the antibacterial activity of the essential oils: Streptococcus mitis (ATCC 49456), Streptococcus mutans (ATCC 25175), Streptococcus sanguinis (ATCC 10556), Streptococcus sobrinus (ATCC 33478), Actinomyces naeslundii (ATCC 19039), Bacteroides fragilis (ATCC 25285), and Prevotella nigrescens (ATCC 33563).
4
Cultivation of Oral Pathogenic Microbes
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The following oral pathogens were studied: Candida albicans CBS 562 from “Centraalbureau voor Schimmelcultures” and bacteria Streptococcus sanguis ATCC 10556, Streptococcus mitis ATCC 903, Porphyromonas gingivalis ATCC 33277 and Fusobacterium nucleatum ATCC 25586 from American Type Culture Collection. The microorganisms were stored at −70°C in Sabouraud Dextrose Broth (SDB, Merck® - C. albicans) and Mueller-Hinton Broth (Difco® - bacteria) with 15% glycerol. It was considered the oxygen exigencies of each microorganism (C. albicans - aerobiosis, S. mitis and S. sanguis microaerophilie and F. nucleatum and P. gingivalis anaerobiosis) to choose bacteria growth conditions.
5
Streptococcus Strain Cultivation and Antibiotic Usage
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The following bacterial strains were used in this study: Streptococcus mutans UA159, Streptococcus sobrinus OMZ175, Streptococcus mitis ATCC 49456, Streptococcus oralis ATCC 10557, Streptococcus gordonii ATCC 10558, Streptococcus sanguinis SK36, Streptococcus parasanguinis ATCC 15912, Streptococcus tigurinus ATCC 15914, Streptococcus australis ATCC 700641, Streptococcus infantis ATCC 700779, Streptococcus salivarius HHT, Streptococcus anginosus NCTC 10707, Streptococcus intermedius ATCC 27335, and Escherichia coli DH5α
E. coli strains were grown in 2×YT Broth (Becton Dickinson, Franklin Lakes, NJ, USA). Oral streptococci strains were grown in brain heart infusion (BHI) broth (Becton Dickinson) at 37°C in 5% CO2. Antibiotics were used at the following concentrations: 300 μg/mL erythromycin and 50 μg/mL ampicillin for E. coli, 20 μg/mL erythromycin for oral streptococci, 600 μg/mL spectinomycin for S. mutans and S. anginosus, and 150 μg/mL spectinomycin for S. sanguinis.
E. coli strains were grown in 2×YT Broth (Becton Dickinson, Franklin Lakes, NJ, USA). Oral streptococci strains were grown in brain heart infusion (BHI) broth (Becton Dickinson) at 37°C in 5% CO2. Antibiotics were used at the following concentrations: 300 μg/mL erythromycin and 50 μg/mL ampicillin for E. coli, 20 μg/mL erythromycin for oral streptococci, 600 μg/mL spectinomycin for S. mutans and S. anginosus, and 150 μg/mL spectinomycin for S. sanguinis.
6
Antimicrobial Activity of Essential Oils
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Bacteria were acquired from the American Type Culture Collection (ATCC) and kept in the culture collection of the Laboratory of Research on Applied Microbiology (LaPeMA) at Universidade de Franca, state of São Paulo, Brazil, at -80°C. The following microorganisms were used: Streptococcus salivarius (ATCC 25975), Streptococcus mutans (ATCC 25175), Streptococcus mitis (ATCC 49452), Streptococcus sanguinis (ATCC 10556) and Streptococcus sobrinus (ATCC 33478).
Minimum Inhibitory Concentrations (MIC) of essential oils were determined by the broth microdilution method in 96-well microplates, following the methodology of CLSI (2006) . Samples were dissolved in 125 µL tryptic soy broth (TSB) to yield compound concentrations between 50 and 400 µg/mL. The inoculum was adjusted to 625 nm for every microorganism in a spectrophotometer to obtain cell concentration of 5 × 10 5 colony-forming units (CFU/mL) (CLSI 2006) . Chlorhexidine digluconate (Sigma-Aldrich), at concentrations from 0.115 to 59.0 µg/mL, was used as the positive control. The microplates were incubated at 37 °C for 24 h; then, 30 µL 0.02% resazurin (Sigma-Aldrich) aqueous solution was added to every well (Sarker et al., 2007) . resazurin is an oxireduction probe that enables microbial growth to be immediately observed. Blue and red colors represent the absence and the presence of microbial growth, respectively.
Minimum Inhibitory Concentrations (MIC) of essential oils were determined by the broth microdilution method in 96-well microplates, following the methodology of CLSI (2006) . Samples were dissolved in 125 µL tryptic soy broth (TSB) to yield compound concentrations between 50 and 400 µg/mL. The inoculum was adjusted to 625 nm for every microorganism in a spectrophotometer to obtain cell concentration of 5 × 10 5 colony-forming units (CFU/mL) (CLSI 2006) . Chlorhexidine digluconate (Sigma-Aldrich), at concentrations from 0.115 to 59.0 µg/mL, was used as the positive control. The microplates were incubated at 37 °C for 24 h; then, 30 µL 0.02% resazurin (Sigma-Aldrich) aqueous solution was added to every well (Sarker et al., 2007) . resazurin is an oxireduction probe that enables microbial growth to be immediately observed. Blue and red colors represent the absence and the presence of microbial growth, respectively.
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