• 제목/요약/키워드: agar diffusion method

검색결과 171건 처리시간 0.026초

Inhibition of Quorum Sensing Regulated Virulence Factors and Biofilm Formation by Eucalyptus globulus against Multidrug-Resistant Pseudomonas aeruginosa

  • Sagar, Pankaj Kumar;Sharma, Poonam;Singh, Rambir
    • 대한약침학회지
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    • 제25권1호
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    • pp.37-45
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    • 2022
  • Objectives: The quorum-sensing-inhibitory and anti-biofilm activities of the methanol extract of E. globulus leaves were determined against clinically isolated multidrug-resistant Pseudomonas aeruginosa. Methods: The preliminary anti-quorum-sensing (AQS) activity of eucalyptus was investigated against a biosensor strain Chromobacterium violaceum ATCC 12472 (CV12472) by using the agar well diffusion method. The effect of sub-minimum inhibitory concentrations (sub-MICs) of the methanol extract of eucalyptus on different quorum-sensing-regulated virulence factors, such as swarming motility, pyocyanin pigment, exopolysaccharide (EPS), and biofilm formation, against clinical isolates (CIs 2, 3, and 4) and reference PA01 of Pseudomonas aeruginosa were determined using the swarm diameter (mm)-measurement method, chloroform extraction method, phenol (5%)-sulphuric acid (concentrated) method, and the microtiter plate assay respectively, and the inhibition (%) in formation were calculated. Results: The preliminary AQS activity (violacein pigment inhibition) of eucalyptus was confirmed against Chromobacterium violaceum ATCC 12472 (CV12472). The eucalyptus extract also showed concentration-dependent inhibition (%) of swarming motility, pyocyanin pigment, EPS, and biofilm formation in different CIs and PA01 of P. aeruginosa. Conclusion: Our results revealed the effectiveness of the E. globulus extract for the regulation of quorum-sensing-dependent virulence factors and biofilm formation at a reduced dose (sub-MICs) and suggest that E. globulus may be a therapeutic agent for curing and controlling bacterial infection and thereby reducing the possibility of resistance development in pathogenic strains.

혈청단백질분획(血淸蛋白質分劃)에 대한 연구(硏究) (I, II) (Studies on Bovine Serum Protein Fractions (I, II))

  • 임봉호
    • 대한수의학회지
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    • 제5권1호
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    • pp.1-16
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    • 1965
  • I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

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여드름 유발균 Propionibacterium acnes에 대한 녹나무과 10종의 항균활성 (Anti-microbial Activities of Ten Lauraceae Species against Propionibacterium acnes)

  • 조주성;지래원;장보국;정헌상;이철희
    • 한국자원식물학회지
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    • 제31권5호
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    • pp.423-432
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    • 2018
  • 본 연구는 녹나무과 10종의 추출물이 Propionibacterium acnes의 억제활성에 미치는 영향을 분석하여 새로운 천연 항균소재를 개발하기위해 수행되었다. 실험재료는 완도와 제주도에서 채집하였으며, 부위(잎, 가지), 용매(증류수, 80% 에탄올, 100% 메탄올) 및 초음파 추출시간(15, 30, 45분)에 따른 조추출물의 항균활성을 agar diffusion method로 조사하였다. 대조구로는 합성항균제인 methylparaben과 phenoxyethanol 0.4, 1, 2, 4 mg/disc를 사용하였다. 연구에 사용된 10종의 모든 추출물에서 억제활성을 보였으며, 여드름균에 대한 항균작용을 확인할 수 있었다. 이 중 월계수는 80% 에탄올에 45분간 추출한 조건에서 20.2 mm의 생육억제환을 나타내어 가장 우수한 여드름균에 대한 항균작용을 확인할 수 있었다. 월계수 잎을 100% 메탄올을 용매로 초음파 30분 추출한 다음n-hexane, chloroform, ethyl acetate, n-butanol 그리고 water 순으로 용매 분획하여 항균활성을 비교분석하였다. 결과, butanol 분획물을 제외한 모든 분획물에서 10.0 mm 이상의 억제환이 형성되었다. 그중 ethyl acetate 분획물에서 억제환이 13.3 mm로 가장 컸으며, 조 추출물(10.2 mm)과 대조구인 phenoxyethanol (12.5 mm)에 비해서도 유의적으로 우수한 억제활성을 나타내었다.

Bifidobacterium spp.와 Clostridium perfringens의 생육에 영향을 주는 식품소재의 탐색 (Screening of the Foodstuffs Influencing the Growth of Bifidobacterium spp. and Clostridium perfringens)

  • 박종현;한남수;유진영;권동진;신현경;구영조
    • 한국식품과학회지
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    • 제25권5호
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    • pp.582-588
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    • 1993
  • 한국인들이 섭취하고 있는 식품이 인체 장내에서 중요한 생리적 기능을 보이는 Bifidobacterium spp.와 Clostridium perfringens의 생육에 미치는 영향을 조사하였다. 수집된 162종의 식품소재 및 식품 추출물 중에서 B. bifidum과 C. perfringens의 생육에 영향을 주는 소재를 agar diffusion method로 31종을 선발하였다. 이들 중에서 알가지, 된장, 양파, 겨자, 감자 등의 추출물은 C. perfringens의 생육을 상하게 억제하였며 Bifidobacterium spp.를 포함하는 기타의 장내미생물의 생육에는 크게 영향을 주지 않았다. 사람 분변을 종균으로 하여 이들 추출물 1%를 첨가하여 배양한 결과 알가지, 양파, 겨자 배양액에서 Bifidobacterium spp.의 생균순는 대조구에 비하여 $10^{2{\sim}3}$개 증가하고 있는 반면 C. perfringens의 생균수는 현저히 감소하였다. 그리고 양파, 겨자의 경우 배양액 중의 ${\beta}-glucosidase$ 역가, indole 함량 등이 현저히 낮아졌고, 양파와 된장에서는 ${\beta}-glucosidase$ 역가가 검출되지 않았다.

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Chemical Composition and Antimicrobial Activity of Essential Oil from Cones of Pinus koraiensis

  • Lee, Jeong-Ho;Yang, Hye-Young;Lee, Hong-Sub;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제18권3호
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    • pp.497-502
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    • 2008
  • The essential oil from the cones of Pinus koraiensis was prepared after removing the seeds, and its chemical composition analyzed using gas chromatography-mass spectrometry (GC-MS). Hydrodistillation of the P. koraiensis cones yielded 1.07% (v/w) of essential oil, which was almost three times the amount of essential oil extracted from the needles of the same plant. Moreover, the antimicrobial activities of the oil against the growth of Gram-positive bacteria, Gram-negative bacteria, and fungi were evaluated using the agar disc diffusion method and broth microdilution method. Eighty-seven components, comprising about 96.8% of the total oil, were identified. The most abundant oil components were limonene (27.90%), ${\alpha}$-pinene (23.89%), ${\beta}$-pinene (12.02%), 3-carene(4.95%), ${\beta}$-myrcene (4.53%), isolongifolene (3.35%), (-)-bornyl acetate (2.02%), caryophyllene (1.71%), and camphene (1.54%). The essential oil was confirmed to have significant antimicrobial activities, especially against pathogenic fungal strains such as Candida glabrata YFCC 062 and Cryptococcus neoformans B 42419. Therefore, the present results indicate that the essential oil from the cones of Pinus koraiensis can be used in various ways as a nontoxic and environmentally friendly disinfectant.

Comparative Analysis of Chemical Compositions and Antimicrobial Activities of Essential Oils from Abies holophylla and Abies koreana

  • Lee, Jeong-Ho;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제19권4호
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    • pp.372-377
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    • 2009
  • The chemical compositions and antibacterial and antifungal activities of essential oils extracted from Abies holophylla and A. koreana were investigated. GC-MS analysis revealed that 38 compounds comprised 95.88% of the A. holophylla essential oil, with the main components being bicyclo[2.2.1]heptan-2-ol(28.05%), ${\delta}3$-carene(13.85%), $\alpha$-pinene(11.68%), camphene(10.41%), dl-limonene(7.61%), $\beta$-myrcene(7.11%), trans-caryophyllene(5.36%), and $\alpha$-bisabolol(3.67%). In the essential oil from A. koreana, 36 compounds comprised 98.67% of the oil, and the main compounds were bornyl ester(41.79%), camphene(15.31%), $\alpha$-pinene(11.19%), dl-limonene(8.58%), fenchyl acetate(5.55%), and $\alpha$-terpinene(2.29%). Both essential oils showed great potential of antibacterial activity against several bacteria tested, in the range of 2.2-$8.8{\mu}g$ per disc by the agar disc diffusion method, and minimal inhibitory concentration(MIC) values of 5.5-21.8 mg/ml by the microdilution method. Both oils showed very effective antifungal activities toward all pathogenic strains tested, including Candida glabrata, with MIC values in the range of 0.5-2.2 mg/ml. As a whole, A. koreana oil showed better antibacterial and antifungal properties than A. holophylla oil.

S. mutans에 대한 대나무 숯의 항균력에 관한 연구 (Antibacterial effect of bamboo charcoal on Streptococcus mutans)

  • 최미숙;안권숙
    • 한국치위생학회지
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    • 제14권1호
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    • pp.95-100
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    • 2014
  • Objectives : The purpose of the present study was to investigate the effect of bamboo charcoal on Streptococcus mutans which is one of the most important causative agents of dental caries. Methods : S. mutans was incubated with or without bamboo charcoal and then changes were observed in its cell viability and antibacterial effect. Oral epithelial cells viabillity(human gingival fibroblast, HGF) was performed using MTT assay. Antibacterial effect was analyzed using a dilution plating method and agar diffusion method. Results : Oral epithelial cells, human gingival fibroblast (HGF) showed a tendency to increase in bamboo charcoal treatment solution concentrations(0.5, 1, 2, 3, 5, 10%). The bamboo charcoal had an antibacterial effect on S. mutans. Antibacterial effect of bamboo charcoal for the bacterium was 58%. Charcoal concentration of 2% and 5% in the inhibition zone showed a minimal growth, but the concentration of 10% bamboo charcoal in inhibition zone revealed a conspicuous antibacterial activity. Conclusions : Overall results suggested that the bamboo charcoal proved to be bactericidal effect on S. mutans.

Estimation of Antibacterial Properties of Chlorophyta, Rhodophyta and Haptophyta Microalgae Species

  • Imran Bashir, Khawaja Muhammad;Lee, Jae-Hyeon;Petermann, Maike Julia;Shah, Abid Ali;Jeong, Su-Jin;Kim, Moo-Sang;Park, Nam-Gyu;Cho, Man-Gi
    • 한국미생물·생명공학회지
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    • 제46권3호
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    • pp.225-233
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    • 2018
  • In this exploratory study, eight types of microalgae from different phyla (Chlamydomonas reinhardtii, Chlorella species, Haematococcus pluvialis, Porphyridium purpureum, Porphyridium cruentum, Isochrysis species, Isochrysis galbana, and Pavlova lutheri) were tested for their antibacterial activities against eight target pathogenic bacterial strains. The agar well diffusion method and broth micro dilution assay were conducted to estimate the antibacterial activity. Microalgae cell-free supernatants, exopolysaccharides (EPS), water, and organic solvent extracts were used for inhibition analysis. EPS extracted from P. lutheri showed activity against Bacillus subtilis and Pseudomonas aeruginosa. Inhibition zone diameters of 14-20 mm were recorded on agar plates, while the minimum inhibitory concentrations in the broth micro dilution assay were $0.39-25mg\;ml^{-1}$. During this study, haptophyte microalgae, Isochrysis species, and P. lutheri extracts showed the highest activity against most of the tested pathogenic bacterial strains, while most of the extracts were active against the important foodborne pathogen P. aeruginosa. This study showed promising results regarding important microalgae phyla, which will further aid research related to extracts and exploitation of bioactive metabolic compounds in the food and pharmaceutical industries.

티벳산 발효유로부터 분리한 Candida kefyr TFP 7의 항균활성 및 항암활성 (Antimicrobial and Antitumoral Activities of Candida kefyr TEP 7 Isolated from Tibetan Fermented Milk)

  • 윤원호;나영미;김창한
    • 한국축산식품학회지
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    • 제23권1호
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    • pp.80-85
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    • 2003
  • 티벳산 발효유로부터 분리한 Candida kefyr TFP7의 항균활성 및 항암활성을 검토하기 위하여 PDA배지를 이용한 십진 희석법에 의해 10균주(TFP1∼10)를 분리하여 몇 종의 병원균과 식중독균, 식품과 관련이 있는 세균, 효모, 곰팡이, 녹조류 및 토양으로부터 분리한 방선균 등 18균주에 대하여 항균활성을 조사하였으며, 또한, 9종류의 인체 암세포주에 대하여 항암활성을 비교하였다. 항균활성 검사에서는 Gram 양성균 중 iwicrococcus iuteus ATCC l1880에 대하여 분리균주 (TFP2∼10)와 방선균인 Styeptomyces murinus JCM 4333에 패하여 분리균주(TFP6∼10)가 뚜렷한 생육억제효과를 나타내었다. 한편, 항암활성 검사에서는 SNU-5(human stomach carcinoma), SW-534(human larynx carcinoma)에 대하여 분리 균주(TFP1∼10) 모두 각각 60%, 70%이상의 생육억제 효과를 나타내었다. 특히, 균주 TFP7은 SNU-5, SW-534에 대하여 각각 77.5%, 76.5%의 가장 우수한 생육억제 효과를 나타내었다.

국내 말에서 분리된 Staphylococcus aureus의 특성 및 약제 내성 양상 (Characteristics and Antimicrobial Resistance Patterns of Staphylococcus aureus Isolated from Horse)

  • 최성균;조길재
    • 생명과학회지
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    • 제18권1호
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    • pp.69-74
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    • 2008
  • 국내에서 사육중인 말에서 채취한 시료로부터 Stapylococcus aureus를 분리 동정한 결과 39.2%(58/148)의 분리율을 나타내었다. S. aureus의 혈장응고효소 생산능 시험에서는 98.3%가 양성반응을 나타내었고 신선 혈장의 희석농도는 1:2에서 가장 높은 양성반응을 나타내었다. 본 연구에서 최종 동정된 S. aureus의 기본적인 생화학 성상은 일반적으로 알려져 있는 S. aureus의 특성과 유사한 결과를 얻을 수 있었으나, urease test의 경우 전체 균주의 73% 만이 양성 결과를 나타내어 균주별로 다양한 반응 양상을 관찰 할 수 있었다. 또한 34종의 substrate에 대한 분해 양상에 대한 검사 결과 fructose, maltose, ${\alpha}$-hydroxy butyric acid, thymidine-5'-mono phosphate, uridine-5'-mono phosphate 등의 substrate에 대해서는 모든 균주가 이용능력을 가지고 있었지만 ribose, sorbitol, xylose, xylitol 등의 substrate에 대해서는 모든 균주가 기질 이용능력을 가지고 있지 않는 것으로 확인되었다. 그리고 PCR에 의한 S. aureus를 동정한 결과 108bp 크기의 band를 확인하였다. 국내 말에서 분리한 58주의 S. aureus에 대한 약제 내성 양상을 조사한 결과 spectinomycin, sulfonamides, erythromycin, tetracyclin, ciprofloxacin, neomycin, enofloxacin, penicillin 등의 약제에 높은 저항성을 나타내었다.