• 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.

• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.46-55
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• 2022

Clostridioides difficile infection (CDI) is a significant cause of hospital-acquired and antibiotic-mediated intestinal diseases and is a growing global public health concern. Overuse of antibiotics and their effect on normal intestinal flora has increased the incidence and severity of infections. Thus, the development of new, effective, and safe treatment options is a high priority. Here, we report a new probiotic strain, Bacillus amyloliquefaciens (BA PMC-80), and its in vitro/in vivo anti-C. difficile effect as a prospective novel candidate for replacing conventional antibiotics. BA PMC-80 showed a significant anti-C. difficile effect in coculture assay, and its cell-free supernatant (CFS) also exhibited a considerable anti-C. difficile effect with an 89.06 ㎍/ml 50% minimal inhibitory concentration (MIC) in broth microdilution assay. The CFS was stable and equally functional under different pHs, heat, and proteinase treatments. It also exhibited a high sensitivity against current antibiotics and no toxicity in subchronic toxicity testing in hamsters. Finally, BA PMC-80 showed a moderate effect in a hamster CDI model with reduced infection severity and delayed death. However, further studies are required to optimize the treatment condition of the hamster CDI model for better efficacy and identify the antimicrobial compound produced by BA PMC-80.

• Korean Journal of Plant Resources
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• v.23 no.5
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• pp.436-444
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• 2010

Methanol extracts of the aerial and rhizome parts obtained from ten species of ferns has been screened for antimicrobial activities against Bacillus subtilis, Escherichia coli, Listeria monocytogenes, Propionibacterium acnes, Streptococcus mutans and Streptococcus sobrinus. Antimicrobial activities were carried out using broth microdilution method and paper disc diffusion assay and the extracts which showed clear zones more than 15mm in concentration of 2 mg/disc were tested for its antimicrobial activities at the $0.125{\sim}2\;mg{\cdot}mL^{-1}$ concentration of each extract for three days. The non-sterilized crude methanol extracts of Osmunda cinnamomea var. fokiensis rhizome showed the highest antimicrobial activities on B. subtilis(39%), E. coli (33%) and L. monocytogenes(58%) at the concentration of $2\;mg{\cdot}mL^{-1}$ after 72 hours. In P. acnes, frond extract of O. cinnamomea var. fokiensis showed most vigorous antimicrobial activities in the all extracts but it showed weak activity(clear zone diameters below 15 mm). All extracts has the antimicrobial activities on Streptococcus, but they exhibited weak activity. At the concentration of $2\;mg{\cdot}mL^{-1}$, only Osmunda japonica rhizome extracts showed 28 and 39% of antimicrobial activities on S. mutans and S. sobrinus after 72 hours and the other extracts showed below 10% of antimicrobial activities on S. mutans and S. sobrinus.

• Korean Journal of Plant Resources
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• v.30 no.4
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• pp.343-351
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• 2017

Plant extracts were screened for antifungal activity against major plant pathogens, Botrytis sp., Collectotrichum sp., Alternaria sp. and Cylindrocarpon sp. using 96-well microdilution method. Among the 662 methanol extracts from 401 plant species, 36 extracts showed complete inhibition of spore germination against at least one of four pathogenic fungi. Extracts of Morus alba twig and Sophora flavescens root showed minimum inhibition concentration (MIC) at $1,250{\mu}g/ml$ against Botrytis sp.. Extracts of Chloranthus japonicus root showed MIC at $1,250{\mu}g/ml$ against Collectotrichum sp.. Extracts of Glycyrrhiza uralensis aerial part, Inula helenium root and Menispermum dauricum root showed MIC between 625 and $1,250{\mu}g/ml$ against Alternaria sp.. G. uralensis aerial part and I. helenium root showed MIC at $1,250{\mu}g/ml$ against Cylindrocarpon sp.. Specifically, the extracts of Agrimonia pilosa root, Angelica tenuissima root, Asarum sieboldii root, Campsis grandifolia leaf and twig, Cnidium officinale root, Dictamnus dasycarpus root, G. uralensis aerial part, I. helenium root and M. alba twig completely inhibited spore germination at lower than $5,000{\mu}g/ml$ against all of four pathogenic fungi. Two methanol extracts from G. uralensis aerial part and M. alba twig may used as a candidate to develop into effective disease management materials in plant cultivation.

• Journal of Life Science
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• v.23 no.9
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• pp.1133-1139
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• 2013

During the collection of boar semen, bacterial contamination usually occurs. The contamination has deleterious effects both on semen quality and on sow fertility. The majority of contaminants are gram-negative bacteria, especially Serratia marcescens. In this study, we developed a PCR assay for the identification of S. marcescens targeting the luxS gene (GenBank no. EF164926). S. marcescens yielded a specific 306 bp PCR product. However, no amplification was observed in the other strains tested. The detection limit of PCR was $50pg/{\mu}l$ of template DNA of S. marcescens. The antimicrobial susceptibility patterns of S. marcescens isolated from boar semen were tested using the disk diffusion method. Gentamicin, ceftiofur, florfenicol, and neomycin showed high sensitivity in this test. The minimum inhibitory concentration (MIC) was also determined by the broth microdilution method. The $MIC_{90}$ values of ceftiofur, enrofloxacin, gentamicin, and neomycin were 8, 8, 8, and $16{\mu}g/ml$, respectively. These results indicate that PCR amplification of the luxS gene is a reliable and effective method for the identification of S. marcescens and that ceftiofur, enrofloxacin, gentamicin, and neomycin are effective semen extenders for controlling S. marcescens.

• Mycobiology
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• v.45 no.1
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• pp.25-30
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• 2017

Metal-based drugs, such as 1,10-phenanthroline, have demonstrated anticancer, antifungal and antiplasmodium activities. One of the 1,10-phenanthroline derivatives compounds (1)-N-2-methoxybenzyl-1,10-phenanthrolinium bromide (FEN), which has been demonstrated an inhibitory effect on the growth of Candida spp. This study aimed to explore the in vitro antifungal activity of FEN and its effect on the membrane integrity of Candida albicans. The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) of FEN against planktonic C. albicans cells were determined using the broth microdilution method according to the Clinical and Laboratory Standards Institute guidelines. Cell membrane integrity was determined with the propidium iodide assay using a flow cytometer and were visualized using scanning electron microscopy (SEM). Planktonic cells growth of C. albicans were inhibited by FEN, with an MIC of $0.39-1.56{\mu}g/mL$ and a MFC that ranged from 3.125 to $100{\mu}g/mL$. When C. albicans was exposed to FEN, the uptake of propidium iodide was increased, which indicated that membrane disruption is the probable mode of action of this compound. There was cells surface changes of C. albicans when observed under SEM.

• Korean Journal of Microbiology
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• v.53 no.3
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• pp.193-199
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• 2017

This study was aimed to evaluate the antimicrobial activity of Penicillium purpurogenum ED76 on several clinically important microorganisms. The endophytic fungus P. purpurogenum ED76 was previously isolated from Swietenia macrophylla leaf. The antimicrobial efficacy of P. purpurogenum ED76 dichloromethane extract was determined via disc diffusion and broth microdilution assay. A kill curve study was conducted and the morphology of extract treated bacterial cells were viewed under scanning electron microscope. The dichloromethane extract showed significant inhibitory activity on 4 test bacteria and 2 test yeasts. The minimal inhibitory concentration of the extract ranged from 125 to $1,000{\mu}g/ml$, which indicates the different susceptibility levels of the test microorganisms to the fungal extract. The kill curve study has revealed a concentration-dependent inhibition for all test microorganisms. With the increase of the extract concentration, the microbial growth was significantly reduced. The scanning electron micrograph of dichloromethane extract-treated Staphylococcus aureus cells showed the total damage of the cells. The cell wall invagination of the bacterial cells also indicates the loss of cellular materials and metabolic activity. The gas chromatography mass spectrometry analysis of the extract also showed that the major compound was stigmasterol, which constitutes 45.30% of the total area. The dichloromethane extract of P. purpurogenum ED76 exhibited significant inhibitory activity on several clinically important bacteria and yeasts. The study proposed a possible mode of action that the extract cause significant damage to the morphology of S. aureus cells.

• International Journal of Oral Biology
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• v.39 no.4
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• pp.201-206
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• 2014

This study investigated the antimicrobial activity of methanol extract of mulberry leaf against 16 strains of mutans streptococci and four species of periodontopathogens: Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans. The antimicrobial activities of the crude extracts or silica gel chromatography fractions of methanol-extracted mulberry leaf were evaluated by determining minimal inhibitory concentrations using an established microdilution method. The cytotoxicity of the extracts of mulberry leaf on KB cells was tested by the methyl thiazolyl tetrazolium assay. Chromatography fraction 12 displayed the most potent antimicrobial activity against all 16 strains of mutans streptococci, P. gingivalis, and P. intermedia. No KB cell cytotoxicity was evident up to $128{\mu}g/ml$ of fraction 12. The methanol extract had no antimicrobial activity against F. nucleatum and A. actinomycetemcomitans. These results suggest chromatography fraction 12 methanol extract of mulberry leaf could be useful in the development of oral hygiene products, such as dentifrice and oral hygiene solution, for the prevention of dental caries.

• Korean Journal of Pharmacognosy
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• v.49 no.2
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• pp.155-164
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• 2018

Nypa fruticans Wurmb is a species of palm, which is widely distributed in the mangrove forest of Southeast Asia. Various parts of N. fruticans has been used as a traditional medicinal plant. However, the physiological activities of N. fruticans has not yet been clarified well. Therefore, in this study, the 50% ethanol extract and its aqueous and ethyl acetate fractions of young shoots of N. fruticans were investigated for their antioxidant, cytoprotective effect, and antimicrobial activities. Every sample possessed very high free radical and various ROS scavenging capacities assessed by employing different in vitro assays such as $DPPH^{\cdot}$, $O_2^{{\cdot}-}$, ${\cdot}OH$, and $^1O_2$ scavenging activities. Based on these results, the cytoprotective effect was investigated using the oxidative hemolysis of erythrocyte. We found that the extract and fractions provide a greater protective effect compared with (+)-${\alpha}$-tocopherol. Furthermore, antimicrobial activities were confirmed against skin pathogens by broth microdilution assay. The ethyl acetate fraction had much higher antimicrobial activities than methyl paraben against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, and Candida albicans. Taken together, our results indicated that the young shoots of N. fruticans may have the potential role as a natural active ingredient through their antioxidant, cytoprotective effect, and antimicrobial activities.

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