• Title/Summary/Keyword: Checkerboard assay

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Antibacterial Activity of Water Soluble Components of Elfvingia applanata Alone and in Combinations with Quinolones

  • Kim, Young-So;Eo, Seong-Kug;Oh, Ki-Wan;Lee, Chong-Kil;Lee, Young-Nam;Han, Seong-Sun
    • Mycobiology
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    • v.29 no.1
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    • pp.11-14
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    • 2001
  • A preparation of water soluble components(EA) was made from carpophores of Elfvingia applanata(Pers.) Karst and its in vitro antibacterial activity on a number of bacterial species was examined by macrobroth dilution assay. Among 16 species of bacteria tested, the most potent antibacterial activity was observed against Staphylococcus epiderrnidis and Proteus vulgaris, of which MICs were 1.25 mg/ml. To investigate the antibacterial effects in combinations of EA with quinolone antibiotics, such as ciprofloxacin, enoxacin, lomefloxacin, norfloxacin, and ofloxacin, the fractional inhibitory concentrations(FICs) and the fractional inhibitory concentration indices(FICIs) for four bacterial strains were determined by macrobroth dilution checkerboard assay. Combinations of EA and quinolones exhibited either additive or indifferent effects of antibacterial activity in most instances. However, both synergistic and antagonistic effects were not observed in any cases.

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Antimicrobial Activity of Elfvingia applanata extract alone and in Combination with Some Antibiotics (Elfvingia applanata 엑스가 수종 항생물질의 항균력에 미치는 병용효과)

  • Kim, Young-So;Rym, Kyo-Hwan;Lee, Chong-Kil;Han, Seong-Sun
    • YAKHAK HOEJI
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    • v.38 no.6
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    • pp.742-748
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    • 1994
  • As part of our search for less toxic antimicrobial agents from natural resources, the carpophores of Elfvingia applanata$(P_{ers}.)K_{ARST}.$ was extracted with hot water. EA, the aqueous extract from the carpophores of E. applanata, was lyophilized and a dark brownish powder was obtained. Antimicrobial activity of EA was tested in vitro against Gram positive and Gram negative bacteria by serial broth dilution method, and the antimicrobial activity was expressed by minimal inhibitory concentration(MIC). Among fourteen species of bacteria tested, the antimicrobial activity of EA was the most potent against Proteus vulgaris showing MIC of 1.250 mg/ml. To investigate the effect of antimicrobial combinations of EA with four kinds of antibiotics(ampicillin, cefazolin, oxytetracycline and chloramphenicol), the fractional inhibitory concentration index(FICI) was determined by checkerboard assay for each strain. The antimicrobial combinations of EA with four kinds of antibiotics resulted in synergism in four instances, but no antagonism was observed. Four instances of synergism were observed when EA was combined with ampicillin against Micrococcus luteus, with cefazolin against Bacillus subtilis, with cefazolin against Micrococcus luteus and with oxytetracycline against Staphylococcus aureus.

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A Study on Antimicrobial Activity of Lysimachia clethroides Duby Root Extracts against Methicillin-resistant Staphylococcus aureus

  • Joung, Dae Ki;Kwon, Dong Yeul;Shin, Dong Won
    • Korean Journal of Plant Resources
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    • v.31 no.6
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    • pp.719-724
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    • 2018
  • Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterium responsible for a number of infections in humans that are difficult to treat, and as a result, is a substantial contributor to morbidity and mortality. In the present study, in search of natural products capable of inhibiting this multidrug-resistant bacterium, we investigated the antimicrobial activity of Lysimachia clethroides Duby root. The antibacterial activities of EtOH extract of Lysimachia clethroides Duby root and its n-hexane, EtOAc, n-BuOH and water fractions were evaluated against 15 strains of methicillin-resistant staphylococcus aureus (MRSA) and 1 standard methicillin-susceptible S. aureus (MSSA) strain by using the minimal inhibitory concentrations (MICs) assay, colorimetric assay using MTT test, checkerboard dilution test. Antimicrobial activity of n-hexane fraction of Lysimachia clethroides Duby root was remarkable. Against the 16 strains, the minimum inhibitory concentrations (MICs) were in the range of $31.25-62.5{\mu}g/ml$ and FICI values for n-hexane fraction of Lysimachia clethroides Duby root+AM and n-hexane fraction of Lysimachia clethroides Duby root+OX were checkerboard method performed using the MRSA, MSSA and one clinical isolate strains via MICI 0.12-1 and 0.25-0.75, showing the increase of synergistic effect. When combined together, these antibiotic effects were dramatically increased. These effective combinations could be new promising agents in the management of MRSA.

Combinatorial effects of arginine and n-hexane extract from Korean red ginseng marc against Streptococcus mutans (Streptococcus mutans에 대한 홍삼박 n-hexane 추출물과 Arginine의 병용 효과)

  • Dong Chung Kim;Man-Jin In
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.337-341
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    • 2022
  • In this study, the effect of the combined use of a lipid-soluble n-hexane extract of red ginseng marc (HERGM) and water-soluble arginine, which exhibits anticaries activity, on the growth of Streptococccus mutans was investigated. As a result of checkerboard assay, HERGM and arginine showed a synergistic effect in inhibiting the growth of S. mutans with a fractional inhibitory concentration index of 0.396. Combination treatments of HERGM and arginine resulted in leakage of nucleic acid components and decrease in the viable cell counts of S. mutans, all of which were proportional to the concentration of HERGM. In conclusion, the synergistic effect of HERGM and arginine on the growth inhibition of S. mutans is mainly attributed to HERGM.

Antimicrobial Activity of Elfvingia applanata Extract Alone and in Combination with Naringenin (Elfvingia applanata 엑스의 항균력 및 Naringenin과의 병용효과)

  • Cheon, Jun-Hee;Lee, Sun-Ok;Rym, Kyo-Hwan;Eo, Seong-Kug;Lee, Chong-Kil;Han, Seong-Sun
    • The Korean Journal of Mycology
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    • v.23 no.2 s.73
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    • pp.153-160
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    • 1995
  • As part of our search for less toxic antimicrobial agents from natural resources, the antimicrobial activity of Elfvingia applanata $(P_{ers.})\;K_{arst.}$ extract was examined alone and in combination with naringenin. EA, the aqueous extract from the carpophores of E. applanata, was lyophilized and a dark brownish powder was obtained. Antimicrobial activity of EA was tested in vitro against nineteen strains of bacteria and eleven strains of fungi by serial broth dilution method, and expressed by minimal inhibitory concentration (MIC). Among nineteen strains of bacteria tested, the antimicrobial activity of EA was the most potent against Proteus vulgaris showing MIC of 1.125 mg/ml. EA also inhibited the growth of the selected fungi at higher concentrations ranging from 7.5 mg/ml to 15.0 mg/ml. To investigate the effect of antimicrobial combinations of EA with naringenin, the fractional inhibitory concentration index (FICI) was determined by checkerboard assay for each strain. The antimicrobial combinations of EA with naringenin resulted in partial synergism against Staphylococcus aureus only, and showed additive effect in two strains including Klebsiella pneumoniae and Salmonella typhi. Antagonism was not found.

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Antibacterial activity of Tonghyeonipal-dan against Methicillin-resistant Staphylococcus aureus (통현이팔단 에탄올 추출물의 Methicillin Resistant Staphylococcus aureus에 대한 항균활성)

  • KIM, In-Won;KANG, Ok-Hwa;KONG, Ryong;KWON, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.30 no.5
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    • pp.15-21
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    • 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.

In Vitro Antibacterial Effects of Wandae-tang Aqueous Extracts and Their Combination Effects with Clindamycin against Gardnerella Vaginalis (완대탕(完帶湯)의 Gardnerella vaginalis에 대한 시험관내 항균력 및 Clindamycin과 병용효과)

  • Lee, Seung-Hye;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.25 no.1
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    • pp.34-46
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    • 2012
  • Objectives: The object of this study was to observe the in vitro antibacterial effects of Wandae-tang extracts and combination of Wandae-tang extracts and Clindamycin against Gardnerella vaginalis ATCC14018. Methods: Antibacterial activities against Gardnerella vaginalis ATCC14018 of Wandae-tang extracts were detected using standard agar microdilution methods. In addition, the effects on the bacterial growth curve were also monitored at minimal inhibitory concentration(MIC) and $MIC{\times}2$ levels. The combination effects of Wandae-tang extracts with Clindamycin were observed by Checkerboard microtiter assay, and the effects of bacterial growth curve was treated with Wandae-tang extracts MIC+Clindamycin MIC, 1/2 MIC and 1/4 MIC, respectively. Results: MIC of Wandae-tang extracts and Clindamycin against Gardnerella vaginalis ATCC14018 were detected as $1.719{\pm}0.856$(0.782~3.125) $mg/m{\ell}$ and $0.010{\pm}0.006$ (0.004~0.016) ${\mu}g/m{\ell}$. In addition, Clindamycin and Wandae-tang extracts were also showed marked dosage-dependent inhibition of bacterial growth, and more dramatical inhibitions were detected in Clindamycin+Wandae-tang extracts MIC treatment. Fractional inhibitory concentration index in combination of Wandae-tang extracts and Clindamycin were detected as $0.294{\pm}0.052$(0.250~0.375) at Checkerboard microtiter assay. Conclusions: The results obtained in this study suggest that Wandae-tang extracts showed antibacterial effects against Gardnerella vaginalis ATCC14018, and they also showed dosage-dependent inhibitory effects on the bacterial growth. In addition, combination treatment of Wandae-tang extracts with Clindamycin showed more synergistically potent inhibitory effects on the growth of Gardnerella vaginalis.

Anti-Aspergillus Activities of Plant Essential Oils and Their Combination Effects with Ketoconazole or Amphotericin B

  • Shin, Seung-Won
    • Archives of Pharmacal Research
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    • v.26 no.5
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    • pp.389-393
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    • 2003
  • The essential oils from Cedrus atlantica, Styrax tonkinensis, Juniperus communis, Lavandula angustifolia, Melaleuca alternifolia, Pelargonium graveolens, Pogestemon patchouli and Rosmarinus officinalis were analyzed by GC-MS. Antifungal activities of the oils were investigated by disk diffusion assay and the broth dilution method against Aspergillus niger and A. flavus. The effects of geraniol and the essential oil fraction from P. graveolens on the antifungal activity of amphotericin Band ketoconazole were examined using a checkerboard microtiter assay against both Aspergillus fungi. Most of the tested essential oils, with the exception of C. atlantica, J. communis, and P. patchouli, significantly inhibited growth of A. niger and to a lesser extent that of A. fIavus, with MICs (minimal inhibitory concentrations) in the range 0.78-12.5 mg/mL. The essential oil fraction of P. graveolens and its main components, geraniol and citronellol, exhibited additive effects with amphotericin B and with ketoconazole against both Aspergillus species, resulting in fractional inhibitory concentration (FIC) indices ranging from 0.52 to 1.00.

Antibacterial Effect of Huanggeumjakyak-tang against Methicillin-resistant Staphylococcus aureus (황금작약탕(黃芩芍藥湯)의 MRSA에 대한 항균활성에 관한 연구)

  • Kim, Esther;Choi, Chonghwan;Kim, Ilhyun;Lee, Hail;Song, Yungsun
    • Journal of Korean Medicine Rehabilitation
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    • v.23 no.3
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    • pp.15-26
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    • 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.

Honokiol as an Effective Antimicrobial Compound against Causative Agent of American foulbrood, Paenibacillus larvae

  • Song, Hyunchan;Kim, Ki-Young
    • Journal of Apiculture
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    • v.34 no.2
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    • pp.131-136
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    • 2019
  • Recently, number of honeybees (Apis mellifera) has visibly decreased because they are vulnerable to some diseases like American foulbrood disease. American foulbrood disease, which is caused by Paenibacillus larvae, is emerged as great cause of decrease in number of honeybees. After antibiotic-resistant strain emerged, it is now more difficult to treat those pathogens successfully. Researches on finding alternative antibacterial compound are ongoing. In this study, we examined the antibacterial effect of honokiol on P. larvae. Honokiol showed great antibacterial effect with minimum inhibitory concentration of 12.5 ㎍/mL and minimum bactericidal concentration of 50 ㎍/mL. An agar diffusion test also confirmed the anti-Paenibacillus larvae activity of honokiol with an inhibitory zone of 9±0.5 mm. Since honokiol is known to interact membrane of some bacteria, we measured 260 nm absorbing particles, which could be induced by leakage of cells, and confirmed that the leakage of P. larvae occurred in dose-dependent manners. However, result of crystal violet assay suggested that honokiol has only mild anti-biofilm formation effect on P. larvae, which means honokiol controls the bacteria by inducing the bursting of membrane. Finally, an additive effect of honokiol with tetracycline and terramycin was found using a checkerboard assay with a fractional inhibitory concentration index value of 0.5.