jbv Journal of Bacteriology and Virology J Bacteriol Virol 1598-2467 2093-0249 Journal of Bacteriology and Virology 10.4167/jbv.2007.37.2.61 jbv-37-61 Original Article Antimicrobial Activity of Extract and Fractions from Drynaria fortunei Against Oral Bacteria JungEun-Kyung ∗ Department of Dental Hygiene, Ulsan College, San 160-1 Hwajeong-Dong, Dong-Gu, Ulsan, 682-715, Korea Corresponding author: Prof. Eun-Kyung Jung. Department of Dental Hygiene, Ulsan College, San 160-1 Hwajeong-Dong, Dong-Gu, Ulsan, 682-715, Korea. Phone: +82-52-230-0798, Fax: +82-52-230-0791, e-mail: ekjung@mail.uc.ac.kr 022007 18022007 37 2 61 68 23052007 20062007 Copyright © 2007 Journal of Bacteriology and Virology 2007 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract

One of the traditional Korean medicine, Drynaria fortunei (D. fortunei) is one of candidates known to be effective for the treatment of inflammation, hyperlipemia, arteriosclerosis, rheumatism, and gynecological diseases such as osteoporosis and bone resorption. The present study investigated the antimicrobial activity of methanol (MeOH) extract and n-butanol (n-BuOH), chloroform (CHCl3), and ethyl acetate (EtOAc) fractions of D. fortunei against oral bacteria. The n-BuOH and CHCl3 fractions (MICs, 0.0078 to 0.3125 mg/ml; MBCs, 0.019 to 0.625 mg/ml) were demonstrated as strong antibacterial activity than the MeOH extract and EtOAc fraction. The combination effects of n-BuOH fraction with ampicillin or gentamicin were synergistic against some oral bacteria. We suggest that D. fortunei could be employed as a natural antibacterial agent in oral care products.

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Time-kill curves of MCs of n-BuOH fraction alone and its combination with MICs of ampicillin or gentamicin against. S. mutans, S. sanguinis, S. sobrinus, S. anginosus, S. criceti, and S. ratti. Bacteria were incubated with n-BuOH fraction along (), n-BuOH fraction with ampicillin (❍), and n-BuOH fraction with gentamicin (▾) over time. Data points are the mean values ± S.E.M. of four experiments. CFU, colony-forming units.

Time-kill curves of MICs of n-BuOH fraction alone and its combination with MICs of ampicillin or gentamicin against S. gordonii, A. actinomycetemcomitans, F. nucleatum, P. intermedia, and P. gingivalis. Bacteria were incubated with n-BuOH fraction along (), n-BuOH fraction with ampicillin (❍), and n-BuOH fraction with gentamicin (▾) over time. Data points are the mean values ± S.E.M. of four experiments. CFU, colony-forming units.

Minimum inhibitory concentrations and minimum bactericidal concentrations (mg/ml) of the extract and several fractions of Drynaria fortunei for some oral bacteria

Strains Drynaria fortunei extract and fractions MICs/MBCs (mg/ml)
MeOH n-BuOH CHCl3 EtOAc Ampicillin Gentamicin
S. mutans ATCC 25175 5/5 0.3125/0.625 0.078/0.156 5/10 4/4×10–3 8/8×10–3
S. sanguinis ATCC 10556 2.5/5 0.3125/0.625 0.078/0.156 5/10 32/32×10–3 8/16×10–3
S. sobrinus ATCC 27607 2.5/2.5 0.019/0.039 0.078/0.156 2.5/2.5 2/2×10–3 4/8×10–3
S. ratti KCTC 3294 5/5 0.0078/0.019 0.156/0.3125 5/10 4/4×10–3 4/8×10–3
S. criceti 1KCTC 3292 2.5/2.5 5/10 0.156/0.3125 2.5/2.5 4/4×10–3 8/8×10–3
S. anginosus ATCC 31412 2.5/2.5 0.0078/0.019 0.019/0.039 2.5/5 4/4×10–3 16/16×10–3
S. gordonii ATCC 10558 1.25/2.5 0.156/0.3125 0.039/0.156 0.3125/0.3125 1/2×10–3 2/4×10–3
A. actinomycetemcomitans ATCC 43717 5/5 1.25/2.5 2.5/5 5/10 64/64×10–3 2/2×10–3
F. nucleatum ATCC 51190 2.5/5 1.25/2.5 1.25/1.25 2.5/2.5 0.25/0.25×10–3 16/32×10–3
P. intermedia ATCC 49046 0.019/0.039 0.019/0.019 0.019/0.039 0.039/0.039 32/32×10–3 0.5/1×10–3
P. gingivalis ATCC 33277 0.625/0.625 0.019/0.039 0.019/0.019 1.25/1.25 0.5/1×10–3 256/512×10–3

Korean Collection for Type Cultures

The MeOH extract and n-BuOH fraction of Drynaria fortunei with or without ampicillin against some oral bacteria

Strains MIC1 MIC (mg/ml) FIC2 (mg/ml) FICI3 Outcome
MeOH n-BuOH MeOH n-BuOH MeOH n-BuOH MeOH n-BuOH
S. mutans Alone 1.25 0.039 0.25 0.125 0.5 0.375 Synergistic Synergistic
ATCC 25175 Ampicillin 1×10–3 0.5×10–3 0.25 0.25        
S. sanguinis Alone 0.625 0.039 0.25 0.25 0.375 0.5 Synergistic Synergistic
ATCC 10556 Ampicillin 4×10–3 2×10–3 0.125 0.25        
S. sobrinus Alone 1.25 0.0045 0.5 0.25 0.623 0.31 Additive Synergistic
ATCC 27607 Ampicillin 0.25×10–3 0.125×10–3 0.123 0.063        
S. ratti Alone 1.25 0.019 0.25 0.5 0.75 1 Additive Additive
KCTC 3294 Ampicillin 2×10–3 1×10–3 0.5 0.5        
S. criceti Alone 0.625 1.25 0.25 0.25 0.75 0.75 Additive Additive
KCTC 3292 Ampicillin 2×10–3 1×10–3 0.5 0.5        
S. anginosus Alone 0.3125 0.019 0.25 0.25 0.75 0.75 Additive Additive
ATCC 31412 Ampicillin 2×10–3 2×10–3 0.5 0.5        
S. gordonii Alone 0.3125 0.019 0.25 0.125 0.5 0.375 Synergistic Synergistic
ATCC 10558 Ampicillin 0.25×10–3 0.125×10–3 0.25 0.25        
A. actinomycetemcomitans Alone 1.25 0.156 0.25 0.063 0.375 0.31 Synergistic Synergistic
ATCC 43717 Ampicillin 8×10–3 4×10–3 0.125 0.25        
F. nucleatum Alone 0.3125 0.078 0.125 0.063 0.625 0.5 Additive Synergistic
ATCC 51190 Ampicillin 0.125×10–3 0.063×10–3 0.5 0.25        
P. intermedia Alone 0.0045 0.0045 0.25 0.25 0.5 0.5 Synergistic Synergistic
ATCC 49049 Ampicillin 8×10–3 4×10–3 0.25 0.25        
P. gingivalis Alone 0.039 0.0025 0.063 0.25 0.313 0.31 Synergistic Synergistic
ATCC 33277 Ampicillin 0.125×10–3 0.063×10–3 0.25 0.063        

The MICs of MeOH extract and n-BuOH fraction of Drynaria fortunei with or without ampicillin against oral bacteria are indicated.

Fractional inhibitory concentration (FIC) was calculated by the combined MICs of the MeOH extract and n-BuOH fraction with or without ampicillin.

Fractional inhibitory concentration index (FICI) = (MIC of combination X + Y) / (MIC of drug X alone) + (MIC of combination X + Y) / (MIC of drug Y alone).

The MeOH extract and n-BuOH fraction of Drynaria fortunei with or without gentamicin against some oral bacteria

Strains MIC1 MIC (mg/ml) FIC2 (mg/ml) FICI3 Outcome
MeOH n-BuOH MeOH n-BuOH MeOH n-BuOH MeOH n-BuOH
S. mutans Alone 1.25 0.019 0.25 0.125 0.75 0.5 Additive Synergistic
ATCC 25175 Ampicillin 4×10–3 2×10–3 0.5 0.25        
S. sanguinis Alone 0.625 0.039 0.25 0.25 0.31 0.31 Synergistic Synergistic
ATCC 10556 Ampicillin 4×10–3 2×10–3 0.5 0.25        
S. sobrinus Alone 0.3125 0.0025 0.5 0.25 1 1 Additive Additive
ATCC 27607 Ampicillin 0.5×10–3 0.5×10–3 0.123 0.063        
S. ratti Alone 0.625 0.009 0.25 0.5 0.75 0.75 Additive Additive
KCTC 3294 Ampicillin 1×10–3 1×10–3 0.5 0.5        
S. criceti Alone 0.625 0.625 0.25 0.25 0.75 0.75 Additive Additive
KCTC 3292 Ampicillin 0.5×10–3 0.25×10–3 0.5 0.5        
S. anginosus Alone 0.3125 0.019 0.25 0.25 0.375 0.375 Synergistic Synergistic
ATCC 31412 Ampicillin 1×10–3 0.5×10–3 0.5 0.5        
S. gordonii Alone 0.156 0.019 0.25 0.125 0.31 0.31 Synergistic Synergistic
ATCC 10558 Ampicillin 1×10–3 1×10–3 0.25 0.25        
A. actinomycetemcomitans Alone 0.625 0.078 0.25 0.063 0.5 0.5 Synergistic Synergistic
ATCC 43717 Ampicillin 0.5×10–3 0.5×10–3 0.125 0.25        
F. nucleatum Alone 0.625 0.039 0.125 0.063 0.5 0.5 Synergistic Synergistic
ATCC 51190 Ampicillin 4×10–3 2×10–3 0.5 0.25        
P. intermedia Alone 0.009 0.0025 0.25 0.25 0.31 0.31 Synergistic Synergistic
ATCC 49049 Ampicillin 0.25×10–3 0.125×10–3 0.25 0.25        
P. gingivalis Alone 0.0045 0.0025 0.063 0.25 0.75 0.5 Additive Synergistic
ATCC 33277 Ampicillin 32×10–3 8×10–3 0.25 0.063        

The MICs of MeOH extract and n-BuOH fraction of Drynaria fortunei with or without gentamicin against oral bacteria are indicated.

Fractional inhibitory concentration (FIC) was calculated by the combined MICs of the MeOH extract and n-BuOH fraction with or without gentamicin.

Fractional inhibitory concentration index (FICI) = (MIC of combination X + Y) / (MIC of drug X alone) + (MIC of combination X + Y) / (MIC of drug Y alone).