• The Korea Journal of Herbology
• /
• v.30 no.5
• /
• pp.15-21
• /
• 2015

Objectives : Methicillin-resistantStaphylococcus aureus(MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts ofTonghyeonipal-dan(THD) which prescription is composed of oriental medicine against MRSA.Methods : The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.Results : The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 μg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism againstS. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 μg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation againstS. aureusshowed synergistic action.Conclusions : Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.

• Korean Journal of Microbiology
• /
• v.55 no.1
• /
• pp.46-51
• /
• 2019

This study was carried out to evaluate antimicrobial effects of a mixture of resin fractionated ethanol extract of Eucalyptus globulus, Yucca recurvifolia, and tea tree (Melaleuca alternifolia). The plant fraction mixture showed low minimum inhibitory concentration (0.24~3.32 mg/ml) against several bacteria and yeast that usually used as the target skin microbes in a cosmetic industry, and it was more effective than antibiotics, triclosan and ampicillin. In a time-kill assay the plant fraction mixture reduced more than 92% of microbial populations during 4 h, and significantly increased leakage of nucleotides from all microorganisms tested. Antimicrobial effect of the plant fraction mixture was not affected by divalent cation ($Mg^{2+}$ and $Ca^{2+}$). These results suggest that the fraction mixture of ethanol extracts of E. globulus, Y. recurvifolia, and M. alternifolia may be utilized as an efficient preservative in cosmetics to prevent contamination by human skin microbes.

• Korean Journal of Microbiology
• /
• v.54 no.3
• /
• pp.272-279
• /
• 2018

This study was carried out to evaluate antimicrobial activity of isolated bacteria from various soils against two strains of Propionibacterium acnes causing acne vulgaris. Among several hundreds of bacterial strains, Paenibacillus elgii DS381, Paenibacillus elgii DS1515, Burkholderia gladioli DS518, and Streptomyces lienomycini DS620 showed high antimicrobial activities against the strains of P. acnes. All isolated bacteria showed 15.5 to 34.3 mm inhibition zone diameter in an agar well diffusion test, and especially DS620 showed the highest inhibition zone diameters (28.3~34.3 mm). Antibacterial substances were expected as lipopeptide (pelgipeptin and paenipeptin) from strains DS381 and DS1515, protease from DS518, and anthracycline antibiotic (daunomycinone) from DS620, and all these showed very low minimum inhibitory concentration [DS381 and DS1515 (0.078 mg/ml), DS518 (0.312 mg/ml), DS620 (0.000078 mg/ml)] against P. acnes. These antibacterial substances could completely kill P. acnes within 24 h in a time-kill assay. These results suggest that antibacterial substances produced by these bacteria may be utilized as useful antimicrobial agent against P. acnes and treatment medicine for acne vulgaris.

• Journal of Korean Medicine Rehabilitation
• /
• v.23 no.3
• /
• pp.15-26
• /
• 2013

Objectives The aim of this study was to investigate antimicrobial activity of Huanggeumjakyak- tang water extract against MRSA. Methods The antibacterial activities of Huanggeumjakyak-tang were evaluated against 3 strains of Methicillin-resistant Staphylococcus aureus (MRSA) and 1 standard Methicillinsusceptible S. aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MICs) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay was performed under dark. Results The MIC (minimum inhibitory concentration) of Huanggeumjakyak-tang water extract against S. aureus strains ranged from 1,000 to $2,000{\mu}g/ml$. So we confirmed that it has a strong antibacterial effect. Also the combinations of Huanggeumjakyak-tang water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. Conclusions The results obtained in this study suggest that Huanggeumjakyak-tang water extract showed antibacterial effect against MRSA, and it also showed reducing effect on the side-effect problems that are the major weak points of traditional antibiotics.

• Journal of Korean Medicine Rehabilitation
• /
• v.25 no.2
• /
• pp.1-13
• /
• 2015

Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (Tris) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.

• Journal of Korean Medicine Rehabilitation
• /
• v.30 no.1
• /
• pp.31-45
• /
• 2020

Objectives In this study, we investigated the antimicrobial activity of a 70% ethanol extract of Maneung-hwan (MEH), which is prescribed by practitioners of oriental medicine for use against methicillin-resistant Staphylococcus aureus (MRSA). Methods The antibacterial activity of MEH against MRSA strains was evaluated using the disc diffusion method, broth microdilution method (minimal inhibitory concentration, MIC), checkerboard dilution test, and time-kill test. The mechanism of action of MEH was investigated by bacteriolysis using detergents or ATPase inhibitors Additionally, mRNA and protein expression were investigated by quantitative reverse transcription-polymerase chain reaction and western blot assay, respectively. Results The MIC of MEH was 25~1,600 ㎍/mL against all the tested bacterial strains. We showed that MEH extract exerts strong antibacterial activity. In the checkerboard dilution test, the fractional inhibitory concentration index of MEH in combination with antibiotics indicated synergism or partial synergism against S. aureus. The time-kill study indicated that the growth of the tested bacteria was considerably inhibited after a 24-h treatment with MEH and selected antibiotics. To measure the cell membrane permeability, MEH (3.9 ㎍/mL) was combined with Triton X-100 (TX) at various concentrations N,N-dicyclohexylcarbodimide (DCCD) was also tested as an ATPase inhibitor. TX and DCCD cooperation against S. aureus exhibited synergistic action. Accordingly, the antimicrobial activity of MEH in the context of cell membrane rupture and ATPase inhibition was assessed. Additionally, the expression of genes and proteins associated with resistance was reduced after exposing MRSA to MEH. Conclusions These results suggest that MEH possesses antibacterial activity and acts as a potential natural antibiotic against MRSA.

• Microbiology and Biotechnology Letters
• /
• v.42 no.4
• /
• pp.361-366
• /
• 2014

In this study, Glycyrrhiza uralensis and Glycyrrhiza glabra extracts, with their countries of origin as Korea (Jecheon), Uzbekistan and China, were prepared under various extraction conditions. There were 8 extraction conditions which the licorice were subjected to, and all conditions had different extraction solvents, temperatures and times. Antimicrobial activity on skin flora was evaluated comparatively by a disc diffusion assay, broth macrodilution assay, and kill time curve assay. Based on the antimicrobial activity of their extract confirmed by disc diffusion assay, we established optimal extraction conditions. The Korean licorice extract (85% ethanol, $40^{\circ}C$, 12 h) showed the best activity amongst the samples examined. In particular, its antimicrobial activity against Propionibacterium acnes was the highest. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of the licorice extracts revealed that the Korean licorice ($156{\mu}g/ml$ and $1,250{\mu}g/ml$) had better antimicrobial activity than that of the Uzbekistani licorice ($625{\mu}g/ml$ and $2,500{\mu}g/ml$) and the Chinese licorice ($625{\mu}g/ml$ and $5,000{\mu}g/ml$). Taken together, it was shown that Korean licorice extracted in group F (85% ethanol, $40^{\circ}C$, 12 h) had the highest antimicrobial activity amongst the licorices from the other countries of origin. These results also suggest that the optimal extraction conditions are 85% ethanol, $40^{\circ}C$, 12 h, and that licorice has a potential application as a natural preservative in cosmetics products, thereby replacing synthetic preservatives.

• ALGAE
• /
• v.26 no.3
• /
• pp.237-241
• /
• 2011

The toxic dinoflagellate Karlodinium veneficum has been implicated in numerous fish kill events around the world. Since this species commonly co-occurs with other morphologically similar dinoflagellates, field monitoring of this species in natural waters via light microscopy only has been problematic. In this study, we investigated temporal changes in K. veneficum's abundance in the waters of Obido, Tongyeong, using a species-specific real-time polymerase chain reaction (PCR) assay. The field survey, from April to December 2010, revealed K. veneficum occurred at low densities (12 to 425 cells $L^{-1}$) during this time and that cell numbers peaked in June (early summer in Korea), indicating this species generally occurs in the warmer season (mostly at $16.9-22.3^{\circ}C$ and 33.4-34.5‰) in the Obido area.

• Journal of the East Asian Society of Dietary Life
• /
• v.22 no.1
• /
• pp.109-119
• /
• 2012

This study analyzed the antibacterial activity of the ethanol extract of Schizandra chinensis Baillon against food pathogenic microorganisms to determine its capabilities as a natural antimicrobial agent. A paper disc diffusion test, minimum inhibitory concentration (MIC) determination, and time-kill assay showed that the ethanol extract strongly inhibits the growth of Listeria monocytogenes, Bacillus cereus, Escherichia coli O157:H7, and Pseudomonas aeruginosa. Release of cytoplasmic ${\beta}$-galactosidase was detected in E. coli, E. coli O157:H7, S. aureus, and P. aeruginosa treated with the ethanol extract. An increase of outer membrane permeability caused by the ethanol extract was also observed. An outward flow of cell constituents was detected in the Gram negative strains treated with the ethanol extract. These results imply that the inner and outer membranes of cells were partially destroyed and cell constituents were released by the treatment of the S. chinensis Baillon ethanol extract. The results of this study indicate that ethanol extract of S. chinensis Baillon evidences a fairly good antibacterial effect.

• The Journal of Internal Korean Medicine
• /
• v.34 no.3
• /
• pp.278-288
• /
• 2013

Objectives : Methicillin-resistant Staphylococcus aureus (MRSA) has a cephalosporin and beta-lactam antibiotic-resistant strains. MRSA is one of the major pathogens causing hospital infection and the isolation ratio of MRSA has gradually increased. Consequently, increased resistance to antibiotics is causing serious problems in the world. Therefore, there is a need to develop alternative antimicrobial drugs for the treatment of infectious diseases. Methods : The antibacterial activities of Ipyo-san were evaluated against 2 strains of MRSA and 1 standard Methicillin-susceptible staphylococcus aureus (MSSA) strain by using the disc diffusion method, minimal inhibitory concentrations (MIC) assay, colorimetric assay using MTT test, checkerboard dilution test and time-kill assay performed under dark. Results : The MIC of Ipyo-san water extract against S. aureus strains ranged from 1000 to $2,000{\mu}g/ml$, so we confirmed that it had a strong antibacterial effect. Also, the combinations of Ipyo-san water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Ipyo-san water extract against MRSA has antibacterial activity so it has potential as alternatives to antibiotic agents. For the combination test, we used Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of Ipyo-san water extract was affected by the cell membrane. Conclusions : We suggest that the Ipyo-san water extract lead the treatment of bacterial infection to solve the resistance and remaining side-effect problems that are the major weak points of traditional antibiotics.