• Title/Summary/Keyword: disc-diffusion assay

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A study on the residues of fluoroquinolones in hen's eggs consumed in Incheon Metropolitan city (인천광역시에서 유통되는 계란의 플루오로퀴놀론계 항균물질 잔류 실태 조사)

  • Park, Eun-Jeong;Lim, Ji-Huen;Lee, Sung-Mo;Im, Jeong-Soo;Oh, Dae-Kyu
    • 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.

Antimicrobial Activities of Methanol Extracts Obtained from Several Ferns (양치식물류의 메탄올 추출물에 항균활성 분석)

  • Shin, So-Lim;Lee, Cheol-Hee
    • 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.

Bioactivity and Chemical Composition of the Essential oil of Tea Tree (Melaleuca alternifolia) (티트리 에센셜오일의 생물활성 및 주요 성분 분석)

  • Yang, Seun-Ah;Jeon, Sang-Kyung;Lee, Eun-Jung;Im, Nam-Kyung;Jung, Ji-Young;Lee, In-Seon
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1644-1650
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    • 2008
  • The essential oil of tea tree (Melaleuca alternifolia) is widely used in traditional Australian medicine for skin lesions and infected injuries. In the present study, we investigated the chemical composition, cytotoxicity and its biological activities. The composition of the oil was analyzed by GC-MS. ${\beta}$-Terpinene (20.87%), ${\alpha}$-pinene (17.60%), p-cymene (11.23%), 3-carene (10.40%), trans-anethole (8.47%) and limonene (4.65%) were the major components in the oil. The results tested by MTT assay indicated that the oil showed no cytotoxic effect, at concentrations up to 5%, for less than 3h. The antiradical capacity was evaluated by measuring the scavenging activity of the essential oil on the 2,20-diphenylpicrylhydrazyl (DPPH) and 2,2'-azino-bis 3-ethyl benzothiazoline-6-sulfonic acid (ABTS) radicals. The oil was able to reduce the both radicals dose-dependently, and the concentration required for 50% reduction ($RC_{50}$) against ABTS radicals ($1.6{\pm}0.02%$) was slightly lower than DPPH radicals ($2.6{\pm}0.29%$). The direct contact and vapor-phase antibacterial activity of the oil were also evaluated using disc diffusion method against Staphylococcus aureus, Streptococcus mutans, Listeria monocytogenes, Acinetobacter baumannii, Escherichia coli, and Vibrio parahaemolyticus. All the Gram-negative bacterial strains tested showed more sensibility to the oil than the Gram-positive strains when compare to the effect of gentamycin. On the other hand, the vapor phase of the essential oil against S. aureus exhibited strongest inhibitory effect.

A Study on the Antibacterial Activity of Combined Administration of Jakyakgamcho-tang and Antibiotics Against MRSA (MRSA에 대한 작약감초탕과 항생제 병용투여의 항균활성에 관한 연구)

  • Dam Hee Kang;Ok Hwa Kang;Hee-Sung Chae;Dong Yeul Kwon
    • Korean Journal of Pharmacognosy
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    • v.54 no.2
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    • pp.72-79
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    • 2023
  • MRSA is Staphylococcus aureus resistant to β-lactam antibiotics, and is a worldwide infectious disease. Even with the discovery of new antibiotics, resistance develops rapidly, so new alternatives are needed. Jakyakgamcho-tang (JGT) is a combination of Jakyak and Gamcho, and has been mainly used as an antispasmodic and analgesic in oriental medicine. This study was conducted to find out whether there is an effect on MRSA in relation to the anti-inflammatory effect of JGT and the antibacterial effect of Jakyak and Gamcho found in previous studies. In this study, in order to investigate the antibacterial activity of JGT and the combined effect of existing antibiotics, after extracting JGT with 70% EtoH, the disc diffusion method, minimum inhibitory concentration (MIC), drug combination effect (FICI), and time-kill analysis (Time-kill assay), metabolic inhibition, Western blot and qRT-PCR analysis were used to confirm the antibacterial activity mechanism of MRSA of JGT. As a result of the experiment, all of MRSA showed antibacterial activity in JGT's disc diffusion method, and the MIC was 250-1000 ㎍/mL. When existing antibiotics and JGT were combined with drugs, most had synergy or partial synergy. In addition, it was confirmed that the degree of bacterial growth was suppressed over time when simultaneous administration for 24 hours. JGT showed a synergistic effect when administered together with the ATPase-inhibitor DCCD, suggesting that it affected the inhibition of ATPase. As a result of observing the expression of PBP2a, and hla protein in the JGT-treated group and the untreated control group through wstern blot, it was confirmed that the protein expression of the JGT-treated group was significantly suppressed, and the expression levels of mecA, mecR1 and hla genes were also suppressed during JGT treatment. was observed by qRT-PCR. Combining the results of the experiment, it can be seen that JGT has antibacterial activity in MRSA, and when combined with existing antibiotics, the effect was increased compared to treatment with the drug alone. This suggests that JGT can be an alternative to treatment for antibiotic resistance of MRSA.

Antioxidant, Antimicrobial and Anti-inflammatory Effect of Boehmeria nivea var. nipononivea Extracts (섬모시풀(Boehmeria nivea var. nipononivea) 추출물의 항산화, 항균 및 항염증 효과에 대한 연구)

  • Jung, Gi Soo;Lee, Sun Hee;Yang, Soo-Kyung;Moon, Sung Pil;Song, Gwanpil;Kim, Ji Young
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.4
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    • pp.339-348
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    • 2020
  • The purpose of this study was to investigate the possible use of the Boehmeria nivea var. nipononivea extract and fractions for the development of natural cosmetic ingredients. The leaves of B. nivea var. nipononivea, extracted by 70% ethanol, were sequentially fractionated with n-hexane, dichloromethane, ethylacetate, and n-butanol. As a result of DPPH and ABTS test, ethyl acetate fractionation was shown to be excellent in radical scavenging activity. For the antimicrobial activities against Staphylococcus aureus, Staphylococcus epidermidis, Cutibacterium acnes and antibiotic resistant strains, MIC and birth control rate were observed by paper disc method. In the antibacterial activity by the disc diffusion assay against S. aureus, S. epidermidis and C. acnes, the dichloromethane and ethylacetate fraction showed stronger antibacterial activity than the other fractions and extract. Moreover, the ethylacetate fraction showed strong nitric oxide (NO) production inhibitory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell. In conclusion, we found that B. nivea var. nipononivea extract was not cytotoxic and showed antioxidant, antimicrobial and anti-inflammatory effects. These results suggest that the Boehmeria nivea var. nipononivea extract and fractions could be applied as an effective cosmetic material with antioxidant activity.

Treatment test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP측정을 이용한 젖소 유방염 치료에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seong-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.393-405
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    • 1989
  • This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

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Antimicrobial Effects against Food-Borne Pathogens and Antioxidant Activity of Rhododendron brachycarpum Extract (만병초 추출물의 식중독 유발균에 대한 항균효과 및 항산화활성)

  • Choi, Moo-Young;Rhim, Tae-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1353-1360
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    • 2011
  • This study was performed to investigate the antimicrobial effects against food-borne pathogens and antioxidant activity of Rhododendron brachycarpum ethanol-extract. The antimicrobial activity of the extract was determined using a paper disc-diffusion method, and the diameter of the clear zone was measured. The diameter of the clear zone in the presence of 10 mg of extract was maximal against Bacillus cereus among the three tested Gram-positive bacteria and against Escherichia coli O157:H7 among the five tested Gram-negative bacteria. Analysis of the minimum inhibitory concentration (MIC) showed that the extract exhibited a similar efficacy as that of sorbic acid, a well-known chemical preservative. The growth inhibitory effects of the extract at concentrations of 250, 500, 1,000, and 2,000 mg/L on food-borne pathogens were determined against Staphylococcus aureus, Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7. Growth of the microorganisms was not affected by the extract at concentrations up to 250 mg/L, but it was significantly (p<0.05) inhibited by the extract at concentrations higher than 1,000 mg/L. The antioxidant effects of the extract were examined via measurement of DPPH radical scavenging activity, inhibition of reactive oxygen species (ROS) generation using fluorescent dichlorofluorescien (DCF) assay, and prevention of peroxyl radical- and hydroxyl radical-induced supercoiled DNA breakage. The $IC_{50}$ of the extract for DPPH radical scavenging activity was about half that of ${\alpha}$-tocopherol, which was used as a positive control. DCF fluorescence intensity decreased as the concentration of the extract increased, demonstrating that ROS generation was inhibited in a concentration-dependent manner. The ROS inhibitory effect of the extract was higher than that of ascorbic acid. The extract prevented supercoiled DNA strand breakage induced by peroxyl radical and hydroxyl radical. Thus, the results of the present study demonstrate that the extract exhibits antimicrobial effects against food-borne pathogens as well as potent antioxidant capacity, suggesting that R. brachycarpum could be used as a natural antibacterial agent and effective antioxidant in food.

Anti-oxidation and Anti-atopic Dermatitis Effect of Herbal Wood Vinegar (한방목초액의 항산화 및 항 아토피 효과)

  • Kim, Tagon;Nho, Hwa Jung;Jun, Sang Hui;Kim, Kang Bae;Kim, Donguk
    • Korean Chemical Engineering Research
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    • v.48 no.6
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    • pp.690-694
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    • 2010
  • In this study, herbal wood vinegar including Bambusoideae, Cinnamomi Cortex, Zingiberis Rhizoma was tested to see possibility for cosmetic or skin related medicine. Anti-oxidation effect of herbal wood vinegar was tested by DPPH free radical scavenging activity, and showed 97% inhibition rate at $50{\mu}g/ml$. Anti-bacterial effect was tested by disc diffusion method, and it indicated strong anti-bacterial activity against normal skin flora Staphylococcus aureus. Whitening effect was measured by tyrosinase inhibition assay, and it was lower compared with vitamin C. Stability test was done by MTT assay, and cell toxicity was relatively high. Stability was also checked, and there was not significant change in color, aroma, appearance and pH during storage. Anti-atopic dermatitis test was done by hairless mouse and herbal wood vinegar recovered damaged skin to almost normal condition after 9 days of application. IgE concentration in herbal wood vinegar treated mouse was also reduced 30% compared with control. From the study, herbal wood vinegar showed good anti-oxidation, anti-bacterial and anti-atopic dermatitis effect, and had promising application in cosmetic or skin related medicine.

Effects of Antibacteria and Adhesive Inhibition of Scutellaria baicalensis Extract on Streptococcus mutans (황금(Scutellaria baicalensis) 추출물에 의한 Streptococcus mutans의 항균 및 부착억제 효과)

  • Paek, Jong-Yoon;Kim, Young-Hyun;Kwon, Hyun-Jeoung;Kim, Eun-Nim;Kim, Wan-Jong;Han, Man-Deuk
    • Journal of dental hygiene science
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    • v.8 no.4
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    • pp.367-373
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    • 2008
  • The natural products are used to be development of new antibacterial substances against human pathogenic bacteria. Adherence to the tooth surface by S. mutans is an important step in initiation of dental caries. This study was to examine antibacterial activity and anti-adhesive effect of Scutellaria baicalensis extract against S. mutans. Extracts of S. baicalensis were tested for antimicrobial activities by paper disc methods and radial diffusion assay methods, and bacterial adherence assay using 3 type of hydroxyapatite. The antibacterial level of ethyl acetate extract, IPK-3 on the growth of S. mutans was 125 mg/ml of minimum inhibitory concentration (MIC). The maximum growth of S. mutans in medium added with IPK-3 extract (50 mg/ml) was delayed to 30 hr, while the highest at 24 hr in control medium. The pH values of the control medium was 5.63 at 18 hr, but the media supplemented with IPK-3 extract was pH 6.50 at 12 hr. In adhesive inhibition assay, S. mutans was labelled with the fluorescent indicator DAPI and measured with fluorescence microscope. Adhesion of S. mutans on hydroxyapatite beads was inhibited by IPK-3 extracts. These results suggest that S. baicalensis extract can be used as an effective material for antibacterial activity and adhesive inhibition against S. mutans.

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Antibacterial Effect of Sinhyowoldosan Against Methicillin-Resistant Staphylococcus aureus (신효월도산(神效越桃散)이 메티실린에 내성이 있는 Staphylococcus aureus에 대한 항균활성에 관한 연구)

  • Shin, In-Sik;Kang, Ok-Hwa;Joung, Dae-Ki;Kang, Hee-Jung;Kim, Ji-Eun;Hwang, Hyeong-Chil;Kim, In-Won;Kwon, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.28 no.1
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    • pp.59-64
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    • 2013
  • Objectives : Methicillin-Resistant Staphylococcus aureus (MRSA) is a cephalosporin and beta-lactam antibiotic-resistant strains. In most cases, that is spread from infected patients and infection rates are growing increasingly. Thus, accordingly, 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 infections diseases. Methods : The antibacterial activities of Sinhyowoldosan were evaluated against 3 strains of Methicillin-resistant staphylococcus aureus(MRSA) and 1 standard Methicillin-susceptible 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 Sinhyowoldosan water extract against S. aureus strains ranged from 500 to 2,000 ${\mu}g/mL$, so we have confirmed it on a strong antibacterial effect. Also, the combinations of Sinhyowoldosan water extract and conventional antibiotics exhibited improved inhibition of MRSA with synergy effect. We suggest that Sinhyowoldosan water extract against MRSA have antibacterial activity, it has potential as alternatives to antibiotic agent. the combination test was used, Triton X-100 (TX) and DCCD for measurement of membrane permeability and inhibitor of ATPase. As a result, antimicrobial activity of SH is affected by the cell membrane were assessed. Conclusion : We suggest that the Sinhyowoldosan 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.