• Title/Summary/Keyword: paper disc-diffusion

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A Study of Antimicrobial Activity of Herbal Extracts on Clostridium difficile (Clostridium difficile에 대한 항균 한약재 탐색 연구)

  • Seong, Eunhak;Lim, Sookyoung;Lee, Myeongjong;Kim, Hojun
    • Journal of Korean Medicine Rehabilitation
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    • v.31 no.1
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    • pp.47-57
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    • 2021
  • Objectives This study was conducted to confirm the possibility of Clostridium difficile infection (CDI) treatment through natural herbal medicines. Methods After screening a total of 77 herbal medicines through the paper disc agar diffusion method, we selected the herbal medicines that showed a effectiveness compared to the positive control vancomycin. Afterwards, drugs that showed inhibitory effects compared to C. difficile without inhibition of Bifidobacterium bifidum and Lactobacillus plantarum, known as beneficial bacteria, were selected and minimal inhibitory concentration (MIC) was confirmed by applying the Broth microdilution method. Results The Coptidis Rhizoma, well known for its antimicrobial effect, was found to have antimicrobial effects on C. difficile, but also had inhibitory effects on the beneficial bacterium B. bifidum. 30% ethanol extraction Crataegi fructus, Corni fructus and Mume fructus had antimicrobial effects on C. difficile without inhibiting the beneficial bacteria B. bifidum and L. plantarum. The MIC values of 30% ethanol extraction Crataegi fructus, Corni fructus and Mume fructus were found to be 10 mg/mL, 20 mg/mL and 5 mg/mL, respectively. Conclusions Crataegi fructus, Corni fructus and Mume fructus were identified as candidate medicines for C. difficile. Further researchs will need to be done in vivo, and to find an optimal extraction method accompanied by economic evaluation.

Antibacterial activity of Bio-fermented Galla Rhios Extract (오배자 발효추출물의 항세균활성)

  • Doh, Eun Soo;Yoo, Ji Hyun
    • The Korea Journal of Herbology
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    • v.29 no.4
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    • pp.21-27
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    • 2014
  • Objectives : This experimental study was performed in order to investigate the antibacterial effect of bio-fermented Galla Rhois extract. Methods : The Galla Rhois extract was fermented by Streptococcus thermophilus, Saccharomyces cerevisiae and Lactobacillus delbrueckii, and their products was tested for antibacterial activity against six pathogenic microorganisms namely, Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Vibrio parahaemolyticus, Escherichia coli and Salmonella typhimurium by paper disc diffusion method. Results : The Galla Rhois fermented extract by Lactobacillus delbrueckii and Saccharomyces cerevisiae showed more effective antibacterial activity than not fermented extract against Bacillus subtilis and Vibrio parahaemolyticus. Antibacterial activity of fermented extract using especially Lactobacillus delbrueckii and Saccharomyces cerevisiae was proved that it was good with even 2 percents concentration. Antibacterial activity of Galla Rhois extract within pH 3 to pH 7 had been safe regardless of pH but low over pH 9. The growth of Bacillus cereus, Staphylococcus aureus, and Vibrio parahaemolyticus had a tendency to decrease depend on the increasing concentration of the extract. EtOEt, EtOAc and n-BuOH fractions of the Galla Rhois extract had a high level of antibacterial activity against Bacillus cereus, Bacillus subtilis, Staphylococcus aureus and Vibrio parahaemolyticus, respectively. Surprisingly, EtOAc fractions of the Galla Rhois extract showed higher antibacterial activity against Vibrio parahaemolyticus alone. And antibacterial activity against six pathogenic microorganisms had a tendency to increase depend on the increasing concentration of the fractions of the Galla Rhois extract. Conclusions : Bio-fermented Galla Rhois extract, efficiently inhibited the growth of Bacillus cereus and Vibrio parahaemolyticus.

Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce (간장에서 분리한 Bacillus licheniformis가 생산하는 박테리오신의 특성 및 정제)

  • Jung, Sung-Sub;Choi, Jung-I;Joo, Woo-Hong;Suh, Hyun-Hyo;Na, Ae-Sil;Cho, Yong-Kweon;Moon, Ja-Young;Ha, Kwon-Chul;Paik, Do-Hyeon;Kang, Dae-Ook
    • Journal of Life Science
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    • v.19 no.7
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    • pp.994-1002
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    • 2009
  • A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

Effects of Artemisia capillaris Extracts on Intestinal Microflora In vitro and In vivo (In vitro 및 In vivo에서 인진쑥 추출물이 장내미생물에 미치는 영향)

  • Oh, Mi-Hyun;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.11
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    • pp.1587-1594
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    • 2010
  • This study was designed to investigate the effect of Artemisia capillaris extracts on the intestinal microflora. In agar diffusion method, the solvent fractions of Artemisia capillaris showed growth inhibition against the intestinal microflora. In particular, the chloroform fraction of Artemisia capillaris had strong antibacterial activity against Clostridium perfringens, Clostridium difficile, Eubacterium limosum, and Bacteroides fragilis, but did not show antibacterial activity against Bifidobacterium bifidum and Lactobacillus acidophilus. Most chloroform fraction of Artemisia capillaris inhibitory activities were not reduced by heat treatment or pH variation against C. perfringens, C. difficile, E. limosum, and B. fragilis. MICs of the chloroform fraction were 1.25 mg/mL against C. perfringens, E. limosum and B. fragilis and 2.5 mg/mL against C. difficile. MBCs of chloroform fraction were 5 mg/mL against C. perfringens, E. limosum and 2.5 mg/mL against C. difficile, B. fragilis. The ethyl acetate fraction of Artemisia capillaris showed $3.08{\pm}0.03$ mg/10 mg total polyphenol and $1.91{\pm}0.03$ mg/10 mg total flavonoid contents. In vivo tests were performed to investigate the influence of Artemisia capillaris extract on the intestinal microflora in rats. The results showed the possibilities of utilizing Artemisia capillaris extracts as a functional food component to control intestinal microflora.

Antimicrobial activity of eight root canal sealers before and after setting (8종 root canal sealers의 경화 전, 후의 항균효과에 관한 연구)

  • Fang, Denny Y.;Lee, Woo-Cheol;Lai, Chern H.
    • Restorative Dentistry and Endodontics
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    • v.27 no.2
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    • pp.207-211
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    • 2002
  • 항균 활성도는 Root canal sealer가 갖추어야 할 필수요소 중 하나이다. 본 연구는 최근 임상술식에 사용되고 있는 8종의 root canal sealer의 근관내 혐기성 세균에 대한 항균효과를 알아보기 위해 시행되었다. 또한 본 연구에서는 혼합직후의 경화되지 않은 sealer와 경화 7일 후 sealer의 항균효과도 비교하였다. 항균효과 측정을 위해 사용된 균주는 최근 실패한 근관치료 증례에서 배양되어 보고된 바 있는 Enterococcus faecalis와 근관내 주요 감염균인 그램음성 혐기성세균인 Staphylococcus aureus를 대상으로 하였고, Agar diffusion test 방법을 사용하였다. 실험방법으로는 2개의 paper disk에 신선하게 혼합한 각각의 sealer를 도포하여 한개의 disk는 즉시 실험에 사용하고 다른 한개의 disk는 일주일간 혐기성 배양기에서 경화시킨 다음 사용한다 각각의 균주를 Brucellar blood agar plate에 접종한 다음, sealer가 도포된 paper disk를 plate상에 올려놓는다. 대조군으로는 식염수에 침윤시킨 disc를 같은 방법으로 각 실험단계에 사용한다. 각 plate를 혐기성 배양기에서 48시간동안 배양한 뒤 실험에 사용한 sealer의 항균효과를 6mm paper-disk를 둘러싼 inhibition zone을 측정하여 평가한다. Fisher's PLSD분석방법 결과 E. faecalis에 대하여 경화 전과 후의 AH26모두 경화 전과 후의 Roth 801, Dentalis, Apexit, AH Plus, RSA그리고 경화 후의 MCS보다 유의성 있게 강한 항균효과를 나타내는 것으로 보고되었으며. 경화 후의 AH26은 경화 전의 AH 26, 경화 전의 Ketac Endo, 경화 전의 MCS보다 통계학적으로 유의성이 있는 항균작용을 하는 것이 관찰되었다 (p<0.05). 경화 후 Roth 801, 경화전과 후의 Dentalis, AH plus, Apexit, RSA는 E. faecalis에 대한 항균효과를 나타내지 못하였다. S. aureus에 대하여 경화후의 AH26이 경화 전과 후의 Roth 801, Apexit, AH Plus, RSA보다 유의성있는 항균효과를 보이는 것을 발견 할 수 있었고, 경화 전의 AH 26이 경화 후의 AH plus보다 나은 항균효과를 나타냄을 알 수 있었다. 또, 경화 전과 후의 Apexit, 경화 후의 AH Plus, 경화 전과 후의 RSA에서는 S. aureus에 대한 항균작용이 발견되지 않았다. 본 실험의 결과 AH26이 가장 강한 항균 작용을 갖는 것을 알 수 있었으며, 각 sealer의 경화 전과 후의 항균효과는 AH26이 경화 전보다 경화 후에서 더 강한 항균효과를 나타내는 것 이외에는 효과의 차이가 없었다.

Adhesion control of Campylobacter jejuni in chicken skin using emulsifiers (유화제를 이용한 계육 표면에서 Campylobacter jejuni의 부착 제어)

  • Oh, Do Geon;Kim, Kwang Yup
    • 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.

Effects of a Blend of Prunus Mume Extract as an Alternative to Antibiotics on Growth Performance, Activity of Digestive Enzymes and Microflora Population in Broiler Chickens (항생제 대체 사료첨가제로서 매실추출 혼합물이 육계의 성장, 소화 효소 활성도 및 장내 미생물 균총에 미치는 영향)

  • Ko, Y.H.;Yang, H.Y.;Kang, S.Y.;Kim, E.S.;Jang, I.S.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.611-620
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    • 2007
  • The current study was designed to define whether a blend of prunus mume extract(25%) containing lactic acid(75%) and grape seed extract(10ppm) could affect in vitro antimicrobial activity and growth performance, intestinal microflora, plasma biochemical profiles and digestive enzymes activities in broiler chickens. In paper disc agar diffusion test, we clearly observed antimicrobial activity against E. coli in response to prunus mume extract or a blend of prunus mume extract. For in vivo test, a total of ninety six 3-d-old male broiler chicks were assigned to basal diet(CON), basal diet supplemented with antibiotics (ANTI) and 0.5% a blend of prunus mume extract(PRNUS) until 35 days of age. Throughout the entire experimental period(3-35 days), there were no differences in BW and FCR between the birds fed the basal diet with antibiotics and the diet supplemented with a blend of prunus mume. However, ANTI group showed a significant increase in BW and total gain compared to CON group. The weights of digestive organs such as the pancreas and mucosal tissues were not affected by dietary treatments. There was no difference in plasma levels of glucose, cholesterol, AST and ALT activity. However, triglyceride in plasma increased(P<0.05) in the birds fed the diet supplemented with 0.5% a blend of prunus mume extract compared to those fed antibiotics supplemented diet. The activities of pancreatic trypsin and amylase, and intestinal hydrolase including disaccharidase were not affected by dietary treatment. The colony forming units(CFU) of lactobacillus in the lower ileal-cecum of the birds fed the diet supplemented with a blend of prunus mume extract was significantly(P<0.05) higher than that of birds fed antibiotic supplemented diet without affecting the CFU of E. coli. In conclusion, the birds fed the diet supplemented a blend of prunus mume as an alternative to antibiotics showed a similar growth performance and an significant increase in lactobacillus population compared with the birds fed basal and antibiotics supplemented diets.

Investigation of Antimicrobial Activity and Stability of Orixa japonica Thunb. Leaf Extract (상산나무 잎 추출물의 항균활성 및 안정성 조사)

  • Choe, Su-Bin;Kang, Sung-Tae
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.39-43
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    • 2014
  • The antimicrobial activity of Orixa japonica Thunb. leaf extract towards 13 microorganism strains was evaluated. Both methanol (MEex) and 70% ethanol extracts showed antimicrobial activity towards Streptococcus mutans, Bacillus cereus, Staphylococcus aureus, and Pseudomonas aeruginosa. MEex showed a higher antimicrobial activity than the 70% ethanol extract. In addition, the dichloromethane fraction (DCMfr) of the MEex also had an antimicrobial effect against the microorganisms examined. The minimum inhibitory concentrations (MICs) towards S. mutans, B. cereus, S. aureus, and P. aeruginosa were 49.22, 24.61, 49.22, and 49.22 mg/mL, respectively. In contrast, the MICs of the DCMfr tpwards S. mutans, B. cereus, S. aureus, and P. aeruginosa were 3.31, 0.21, 1.7, and 1.7 mg/mL, respectively. The MEex antimicrobial activity was not affected by a 3 h exposures to pH in the range of 3-11 or by temperatures were maintained between $80^{\circ}C-100^{\circ}C$ for 6 h. However, the MEex antimicrobial activity decreased at a heat treatment of $121^{\circ}C$ 1 h.

Antimicrobial Effects of Herbal Medicine Extracts on Staphylococcus aureus and Escherichia coli O157:H7 (생약재 추출물의 Staphylococcus aureus 와 Escherichia coli O157:H7에 대한 항균효과)

  • Cai, Hua;Choi, Soo-Im;Lee, Yun-Mi;Heo, Tae-Ryeon
    • KSBB Journal
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    • v.17 no.6
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    • pp.537-542
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    • 2002
  • To screening of antimicrobial activity, 95% ethanol and hot water extracts of roots, fruits, leaves, radix and stems of 50 species of traditional herbal medicines were examined. For their growth inhibitory effects on two food-born microorganisms, S. aureus and E. coli O157:H7, by the paper disc diffusion method and the minimum inhibitory concentration(MIC) test. Moutan radicis Cortex and Achyranthis Radix showed the highest inhibitory activities on both S. aureus and E. coli O157:H7. The Inhibition zones of Moutan radicis Cortex on S. aureus and E. coii O157:H7 were 22 mm and 24 mm respectively, and the corresponding inhibition zone of Achyranthis Radix were 23 mm and 22 mm. The MIC or Achyranthis Radix on S. aureus was 156.25 $\mu$g/mL, and the MIC or Achyranthis Radix and Moutan radicis Cortexas on E. coli O157:H7 were 625 $\mu$g/mL and 312.5 $\mu$g/mL, respectively. Their antimicrobial activities in ethanolic extracts were significantly higher than in hot water extracts. In the various solvent fractions prepared from ethanol extract, the ethyl acetate fraction of Achyranthis Radix and the CHCl$_3$ fraction of Moutan radicis Cortexas showed strongest activity.

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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