• Journal of Life Science
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• v.34 no.6
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• pp.393-398
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• 2024

The purpose of this study was to verify the antifungal properties of various herbal medicines belonging to the Ranunculaceae family and to find an extraction method effective in inhibiting fungal growth. When antifungal activity was measured in a liquid medium with extracts obtained by either hot water extraction or organic solvent extraction of the herbal medicines Clematis apiifolia, Coptis chinensis, and Pusatilla chinensis, effective results were obtained from the chloroform extract. In addition, fungal growth inhibition experiments were performed on unicellular fungi, Candida albicans, Candida tropicalis, and Candida lusitaniae, and on filamentous fungi, such as Pythium ultimum, Aspergillus fumigatus, and Fusarium oxysporum, using disk diffusion experiments on solid media. It was confirmed that P. chinensis extract has excellent antifungal properties against Candida spp. and C. apiifolia extract against filamentous mold. Finally, GC-MS analysis was performed to explore the useful antifungal substances present in the extract. As a result of the study, thurbergenone from C. apiifolia and 16-hydroxycleroda-3, 13(14)-dien-15, 16-olide (16-HCDO) from C. chinensis were confirmed as antifungal candidates. In conclusion, it was confirmed that C. apiifolia, C. chinensis, and P. chinensis have antifungal activity against various fungi, and in GC-MS analysis, all herbal medicines were confirmed to have different antifungal candidates. These results indicate that the Ranunculaceae family has evolved in several directions for fungal resistance traits.

• Korean Journal of Veterinary Service
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• v.31 no.3
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• pp.385-395
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• 2008

This study was carried out to investigate the residue level of fluoroquinolones in hen's general eggs and specific eggs by microbiological assay method and high performance liquid chromatography (HPLC) method. HPLC separation was carried out by reversed phase chromatography on a Symmetry $C_{18}$ (250${\times}$4.6 mm, $5{\mu}m$ particle size) with a phase composed of distilled water (containing 0.4% triethylamine and phosphoric acid) : Methanol (780 : 220, v/v), pumped isocratically at a flow rate of 1.0ml/min. A fluorescence detector was utilized with an excitation wavelength of 278nm and an emission wavelength of 456nm. The calibration curves were linear $({\gamma}^2{\geq}0.999)$ over a concentration range of $0.025{\sim}0.4{\mu}g/ml$. Average recoveries of the five fluoroquinolones in whole eggs at fortified levels of $0.05{\sim}0.2{\mu}g/g$ were ranged mean $78.1{\sim}91.7%$ and low coefficient of variation was less than 10% for all analysed samples. The limits of detection and limits of quantification for whole eggs were $1.2{\sim}6.0ng/g$ and $2.3{\sim}9.1ng/g$, respectively. Only one hen's general eggfrom chicken farm in Incheon was detected with the residual fluoroquinolones (Microbiological assay method; 1 of 47 general eggs) ; the range of residual concentration enrofloxacin was 0.12ppm. Those in food stores were detected with the residual fluoroquinolones (Microbiological assay method; 4 of 88 general eggs) ; the ranges of residual concentration enrofloxacin were $0.15{\sim}2.2 ppm$, ciprofloxacin $0.01{\sim}0.06ppm$, and hen's specific eggs (40) in food stores were not detected. For the microbiological assay method of fluoroquinolones in hen's eggs, as the results of comparative analysis, the disc diffusion method with E coli may be a little highly detected for the residual fluoroquinolones.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.334-341
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• 2004

Polysaccharides were prepared from Orostachys japonicus by extration with hot steam water (OJPl). The OIPl fraction was further purified by Sephadex G-50 gel filtration chromatography to produce FI (polysaccharides) and FII (oligosaccharides) fraction. The average molecular masses o fFI and FII fraction were determined to be 3050 kDa and 13 kDa, respectively. The antimicrobial activity of OIPl was tested against 8 strains of bacteria and one strain of yeast by the disc diffusion method, fluorescein diacetate (FDA) method and broth dilution method. The OIPl exhibited a very strong growth inhibition to Candida albicans. The OIPl remarkably sup­pressed the growth of Salmonella typhimurium and Staphylococcus aureus. The OIPl showed higher growth inhibition to Escherichia coli and Pseudomonas aeruginosa than propolis, positive control. When the anticancer activity of the OIPl, FI or FII was examined against human cancer cell lines and the Sarcoma 180 cells, these widely suppressed the proliferation of cell lines in the MTT assay and morphology study. Especially, they remarkably inhibited the growth of A549, HeLa and AGS cells. Also treatment of cancer cells with OJPl, FI or FII induced apoptotic cell death characterized by DNA fragmentation. The OJPl, FI or FII exhibiting various biological activities such as antimicrobial activity and anticancer activity is expected to be developed as new biohealth products.

• Journal of Food Hygiene and Safety
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• v.30 no.1
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• pp.51-58
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• 2015

Thirty one of Staphylococcus aureus isolated from perilla leaf cultivation areas in Miryang were investigated on the characteristics, such as enterotoxin genes and antibiotic susceptibility. Five toxin genes (sea, seb, sec, sed, and see) were examined by PCR method. Disc diffusion method was used to examine the antibiotic susceptibility of S. aureus by using 18 types of antibiotic discs with different concentrations. Among enterotoxin-encoding genes, sea and sed genes were co-detected from 4 isolates (12.9%), sed gene was founded in 9 isolates (29.0%), and see gene was founded in 1 isolate (3.2%). However seb and sec and tsst were not detected in any isolates. As a result of antibiotic susceptibility test, 7 isolates (22.6%) were resistant to 12 antibiotics (penicillin, ampicillin, oxacillin, amoxicillin-clavulanic acid, cefazolin, cephalothin, imipenem, gentamicin, tetracycline, ofloxacin, norfloxacin, and erythromycin). 2 isolates (6.5%) were resistant to 5 antibiotics (penicillin, ampicillin, amoxicillin-clavulanic acid, gentamycin, and telithromycin). MRSA (Methicilline Resistant Staphylococcus aureus) was founded in packing vinyl, hands, and perilla leaves.

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

• The Korea Journal of Herbology
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• v.31 no.5
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• pp.93-98
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• 2016

Objectives : Hwangheuk-san is a complex prescription composed of oriental traditional medicine and has been reported for antioxidant, antimicrobial and anticancer effects in the recent study. Methicillin-resistant Staphylococcus aureus (MRSA) is one of important causes of fatal infectious diseases such as septicemia, endocarditis, toxic shock syndrome, pneumonia, skin and soft tissue infections (SSTIs). S. aureus is reported as being for a variety of human diseases and its epidemiological relevance is mainly due to their ability of becoming highly resistant to common antimicrobials such as tetracycline, penicillin, cphalosporin and aminoglycoside. The objective of this study is to determine the antimicrobial effect of Hwangheuk-san ethanol extracts (HHS) and synergistic effects with antibiotics oxacillin against MRSA.Methods : The antimicrobial activity of HHS was measured by the disc diffusion method, broth microdilution method and the checkerboard dilution test, time-kill curve assay was performed to investigate synergistic effects with antibiotics oxacillin against MRSA.Results : HHS showed antimicrobial activity against MRSA with a MIC value of 125 ㎍/㎖. In the checkerboard test, the interaction of HHS with antibiotics oxacillin produced almost synergy or partial synergy against MRSA. This study showed that HHS reduced the MICs of oxacillin tested, and a remarkable antibacterial effect of HHS, with membrane permeability enhancers.Conclusions : These results suggest that HHS has the antimicrobial effect and synergistic effects with antibiotics oxacillin against MRSA. This study thus can be a valuable source for the development of a new drug with low MRSA resistance.

• The Journal of the Korean Society for Microbiology
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• v.22 no.3
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• pp.309-321
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• 1987

For the investigation of microbiological and immunological specificity of Bacteroides gingivalis, Bacteroides gingivalis were isolated, enumerated and characterized from 13 Korean rapidly progressive periodontitis and 7 healthy control by anaerobic culture technique. The total proportion of black-pigmented Bacteroides from Korean R.P.P. patients and healthy control were 8.78% and 0.92%, respectively, among total isolated black-pigmented Bacteroides. In antibiotic susceptibility test, Bacteroides gingivalis isolated from R.P.P. patients were sensitive to Ampicillin and Tetracycline, and resistant to Gentamicin and Erythromycin in disc diffusion method. In antibiotic broth dilution method, the minimum inhibitory concentration(MIC) to Bacteroides gingivalis was 2 unit/ml of Penicillin and $0.25{\sim}1{\mu}g/ml$ of Tetracycline, respectively. The concentration of serum IgG in rapidly progressive periodontitis patients were sigificantly higher than that of healthy control, and concentration of diluted gingival crevicular IgG has not any significant differences between two groups. Serum and gingival crevicular IgG antibody to Bacteroides gingivalis were significantly higher titer in rapidly progressive periodontitis patients to compare with healthy control. The lipopolysaccharide profiles of 2 Korean B. gingivalis in silver stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis were similar to type strains of B. gingivalis and typical LPS band were appeared around the 24-Kd molecular weight. Immunodiffusion test and immunoelectrophoresis of the L.P.S. extracted from 2 Korean B. gingivalis and 2 kinds of type strains of B. gingivalis showed that B. gingivalis Korean-1 was reacted identically to B. gingivalis ATCC 33277. In trypsin and ${\alpha}$-glucosidase activity test of 2 Korean B. gingivalis, both of them revealed positive trypsin and negative ${\alpha}$-glucosidase activity, respectively. These investigation suggested that B. gingivalis is important pathogenic plaque bacteria for the pathogenesis of periodontitis and further study is needed to purify and characterize of the species-specific antigens of this organisms to develop monoclonal antibody and potential diagnostic reagents.

• Korean Journal of Food Science and Technology
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• v.52 no.6
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• pp.670-677
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• 2020

To prevent contamination by Campylobacter jejuni during chicken carcass processing, the effect of emulsifiers on C. jejuni inoculated on chicken skin was investigated using confocal laser scanning microscopy. Among the 8 emulsifiers (SWA-10D, L-7D, M-7D, S-1670, L-1695, P-1670, polysorbate 20, polysorbate 80) tested for antimicrobial activity by the paper disk method, 4 emulsifiers (L-7D, L-1695, polysorbate 20, polysorbate 80) were screened further. Emulsifier L-1695 showed the largest clear zone at a concentration of 200 mg/mL. The 4 emulsifiers subjected to primary screening were screened for heat and pH stability. In the contact surface test, emulsifier L-1695 showed the lowest log CFU/㎠ value on both stainless steel and ceramic surfaces. When emulsifier L-1695 was applied via general and electrostatic spray methods, the number of C. jejuni entrapped inside chicken skin follicles was significantly reduced in both methods. In conclusion, the emulsifier L-1695 could be employed as a microbial detachment agent in the chicken carcass processing industry.

• Clean Technology
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• v.29 no.2
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• pp.109-117
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• 2023

The purpose of this study is to develop a method for separating antioxidants and sugars from grape skin extract. The extract was first mixed with a variety of organic solvents to investigate whether the separation was feasible. When employing acetone, ethanol, dimethylsulfoxide, or dimethylformamide, the organic solvent-extract combination formed a single phase. However, when benzene, ethyl acetate, or n-hexane was added to the extract, the mixture separated into an organic and an aqueous phase and the pigments remained in the aqueous phase. On the other hand, when polyethylene glycol-2,000 (PEG-2000) and sodium citrate were added to the extract, the mixture was separated into three layers, with the majority of the flavonoids migrating to the top layer and 53% of the extract's glucose migrating to the bottom layer. The top layer had significant antioxidant activity, whereas the bottom layer showed no antioxidant activity. The glucose recovery in the bottom layer increased as the molecular weight of PEG increased and the highest recovery (67%) was observed when PEG-8,000 was added. The highest flavonoid separation was observed with PEG-2,000, followed by PEG-8,000 and PEG-400. The flavonoid separation when PEG-2,000 was added resulted in a flavonoid recovery of 48% and 0.2% from the top and bottom layers, respectively. Examining the effect of the separated solution using the agar disc diffusion method on yeast cell growth confirmed that the addition of the extract, the top, and the bottom layer did not inhibit cell growth.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.8
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• pp.1144-1149
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• 2015

The aim of this study was to investigate the antioxidant and antimicrobial activities of various solvent (acetone, ethyl acetate, and ethanol) extracts from Lentinus edodes. The antioxidant activities were evaluated by measuring total polyphenol and flavonoid contents, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity. Total polyphenol content and ABTS radical scavenging activity were highest in ethanol extract. ABTS radical scavenging activity of ethanol extract showed the highest value (98.5%), which was similar to that of ascorbic acid (95.7%). The ethyl acetate extract from Lentinus edodes showed relatively high total flavonoid content and DPPH radical scavenging activity. Negative correlations were found between total polyphenol contents and DPPH radical scavenging activities in Lentinus edodes extracts. Antimicrobial activities of the extracts were determined against Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus, Escherichia coli, Pseudomonas aeruginosa, and Enterobacter cloacae by the disc diffusion method. The acetone and ethanol extracts showed moderate antimicrobial activities against almost all tested microorganisms except E. coli and S. aureus, respectively. The ethyl acetate extract showed a significant growth inhibition effect against E. coli, Ent. cloacae, and B. subtilis.