• 제목/요약/키워드: Disc-diffusion method

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Evaluation of Cumulative and Conditional Antibiotic Release from Vancomycin-Embedded Fibrin Sealant and Its Antibacterial Activity : An In Vitro Study

  • Shin, Dong-Won;Sohn, Moon-Jun;Cho, Chong-Rae;Koo, Hae-won;Yoon, Sang Won
    • Journal of Korean Neurosurgical Society
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    • 제63권1호
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    • pp.45-55
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    • 2020
  • Objective : Fibrin sealants have been used for hemostasis, sealant for cerebrospinal fluid leakage, and adhesive barrier in neurosurgery. Further, as its clinical use and role of an effective drug delivery vehicle have been proposed. This study was performed to measure antibacterial activity and continuous local antibiotic release from different concentrations of vancomycin-impregnated fibrin sealant in vitro. Methods : Antibacterial activity was investigated by disk diffusion test by measuring the diameter of the growth inhibition zone of bacteria (methicillin-resistant Staphylococcus aureus, ATCC29213) from vancomycin-embedded fibrin sealant disc diluted at five different concentrations (C1-C5; 8.33, 4.167, 0.83, 0.083, and 0.0083 mg/disc, respectively). Continuous and conditioned release of vancomycin concentration (for 2 weeks and for 5 days, respectively) were also measured using high-performance liquid chromatography (HPLC) method. To mimic the physiologic wound conditions with in vitro, conditioned vancomycin release in phosphate buffer solution (PBS) was measured and replaced PBS for five consecutive days, half a day or completely daily. Results : In the disk diffusion test, the mean diameters of bacterial inhibition zone were 2.54±0.07 cm, 2.61±0.12 cm, and 2.13±0.15 cm (C1, C2, and C3 respectively) but 1.67±0.06 cm and 1.23±0.15 cm in C4 and C5, respectively. Continuous elution test elicited the peak release of vancomycin from the fibrin sealant at 48 hours, with continued release until 2 weeks. However, conditioned vancomycin release decreased to half or more on day 2, however, the sustainable release was measured over the therapeutic dose (10-20 ㎍/mL) for 5 days and 4 days in assays of half and total exchange of PBS. Conclusion : This study suggests that fibrin sealant can provide an efficient vehicle for antibiotic drug release in a wide range of neurosurgical procedures and the safe and effective therapeutic dose will be at the concentration embedded of 4.167 mg/disc or more of vancomycin.

가자, 오배자, 계피 추출물을 이용한 화장품 제형에서의 방부효과 (Antimicrobial Plant Extracts as an Alternative of Chemical Preservative: Preservative Efficacy of Terminalia chebula, Rhus japonica (gallut) and Cinnmomum cassia Extract in the Cosmetic Formular)

  • 조은미;배준태;표형배;이근수
    • 대한화장품학회지
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    • 제34권4호
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    • pp.325-331
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    • 2008
  • 본 연구는 천연 식물 추출물을 이용하여 화장품 제형 내에서 천연 방부제로서 효능을 확인하기 위한 목적으로 진행하였다. 선별된 15종의 천연 식물 추출물을 이용하여 병원성 세균(Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli)과 효모균(Candida albicans), 곰팡이균(Aspergillus niger)에 대한 paper disc법(disc diffusion method)과 최소저해농도측정법(MIC)을 이용하여 항균 활성을 측정하였다. 그 결과 가자, 오배자 추출물은 그람 양성인 S. aureus와 그람 음성균인 P. aeruginosa, E. coli 에 대한 항세균 활성이 우수하였고, 계피 추출물은 효모균인 C. albicans와 곰팡이균인 A. niger에 대해 우수한 항진균 활성을 나타냈다. 최종 선별된 3종 외 식물추출물은 효과가 없었다. 항균 활성 효과가 우수한 최적 조합비로 구성된 가자, 오배자, 계피 추출물의 혼합 추출물을 화장품 제형에 첨가하여 제형 내 방부 활성을 측정한 결과 상업적으로 널리 사용되고 있는 파라벤 혼합 방부제와 유사한 정도의 방부 활성을 나타냄을 확인하였다. 결론적으로, 화학 방부제를 대신하여 천연 식물 추출물로만 이루어진 넓은 항균 스펙트럼을 갖춘 방부 시스템의 개발이 가능할 것으로 사료되며 최근 다양한 문제가 제기되고 있는 화학 방부제의 단점을 극복한 천연 방부제를 함유한 화장료의 개발에 응용 가능할 것으로 기대된다.

Isolation of Streptococcus thermophilus and Lactobacillus delbrueckii as Starter Culture Candidate Originated from Indonesian Cow's Milk

  • Andrian, Danish;Rizkinata, Denny;Susanto, Tan Steven Ryan;Lucy, Jap;Jan, Tan Tjie
    • 한국미생물·생명공학회지
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    • 제46권3호
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    • pp.201-209
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    • 2018
  • Streptococcus thermophilus, Lactobacillus delbrueckii, Lactobacillus fermentum and Lactobacillus casei were successfully isolated from indigenous Indonesian fresh milk based on the general morphological and biochemical classification as described in Bergey's manual. Verification was conducted by sequencing of 16S rRNA after selection using the classification method mentioned in the manual. All isolates exhibited antimicrobial activity against Escherichia coli and Staphylococcus aureus in the well diffusion test. The susceptibilities of the isolated S. thermophilus 24/S1 and L. delbrueckii 94/L4 against 22 different antibiotics were determined by the disc diffusion method and variable susceptibility patterns were observed. Both isolates were susceptible to amoxicillin, the most commonly prescribed antibiotic, and resistant to sulfonamide. The presence of a plasmid was not detected after extraction. S. thermophilus 24/S1 and L. delbrueckii 94/L4 starter cultures were prepared for yogurt production after 9.5 h of incubation and the yogurt was evaluated for its flavor and quality by 30 volunteers. A score of $4.93{\pm}0.45$ out of 7 was obtained as compared to the yogurt prepared using commercial starter cultures which yielded a score of $4.76{\pm}0.30$ out of 7.

Screening of New Bioactive Materials from Microbial Extracts of Soil Microorganism (I) Antimicrobial Activity from 200 Sampled Using Microdilution Assay

  • Jung, Sang-Oun;Kim, Joon;Chang, Il-Moo;Ryu, Jae-Chun
    • Archives of Pharmacal Research
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    • 제21권3호
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    • pp.278-285
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    • 1998
  • The microdilution assay recommended by NCCLS (National Committee for Clinical Laboratory Standards) is one of the standardized methods of antibiotic susceptibility test. This method has been widely used clinically to obtain MIC values of antibiotics on pathogenic microorganisms. It is more convenient, rapid and simple to test many samples than other test methods such as agar diffusion assay and broth macrodilution assay. The screening of antimicrobial agents from microbial extracts is too laborious in its process. Therefore, a number of screening methods having more simple procedure have been developed. In our laboratory, we applied microdilution assay for screening the antimicrobial agents. This assay showed dose-response results and was more sensitive than disc diffusion assay in our system. We tested 200 samples of microbial extracts originated from 100 microbial strains and selected several samples as potential candidates. In this report, we show that the microdilution assay is more convenient method in screeing of antibiotic susceptibility than those previously reported.

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Effect of various chromatographic terpenoid fractions of Luffa cylindrica seeds on in-vitro antimicrobial studies

  • Nagarajan, K.;Saxena, Pallavi;Mazumder, Avijit;Ghosh, L.K.;Devi, G. Uma
    • Advances in Traditional Medicine
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    • 제10권1호
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    • pp.21-28
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    • 2010
  • The objective of the present investigation is to evaluate the antimicrobial potency of the terpenoid fractions isolated from Luffa cylindrica seeds against various pathogenic microbes. The seeds were powdered and extracted with methanol in soxhlet appratus based on phytochemical screening. Three terpenoid components were isolated by column chromatography and identified by thin layer chromatography and chemical analysis which were designated as ${LCSF_4}^*$, ${LCSF_6}^*$ & ${LCSF_8}^*$ respectively. Disc diffusion method was employed to determine the antimicrobial effectiveness of test compounds I, II and III $({LCSF_4}^*,\;{LCSF_6}^*\;&\;{LCSF_8}^*)$ against 6 microbial species viz., Staphylococcus (S.) aureus, Bacillus (B.) subtilis, Escherichia (E.) coli, Pseudomonas (P.) aeruginosa, Candida (C.) albicans and Aspergillus niger. The disc was saturated with $100{\mu}l$ of each compound, allowed to dry and introduced on the upper layer of seeded agar plate. The plates were incubated overnight at $37^{\circ}C$. Microbial growth was determined by measuring the zonal inhibition diameters. Compound I showed maximum potency against gram positive S. aureus (21 mm) in comparison with standard ciprofloxacin (38 mm), whereas the same compound was completely devoid of activity against both the fungi tested. Compound II was found to be highly sensitive against both the gram negative E. coli (20 mm) and P. aeruginosa (22 mm). Compound II was found to exhibit maximum potency against the fungi C. albicans (15 mm) and A. niger (20 mm). Compound III was found to be very effective against both the gram positive S. aureus (20 mm) and B. subtilis (15 mm) respectively.

Antibacterial and anti-obesity effects of Abeliophyllum distichum Nakai: an in vitro study

  • Song, Dong Cheol;Lee, Ji Hwan;Oh, Han Jin;Kim, Yong Ju;An, Jae Woo;Chang, Se Yeon;Go, Young Bin;Cho, Hyun Ah;Cho, Jin Ho
    • 농업과학연구
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    • 제48권3호
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    • pp.557-565
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    • 2021
  • Interest in research on various medicinal plants has increased globally over the last few decades, possibly due to their possible antibacterial and antioxidant activities. The present study was conducted to verify the antioxidant effects, antibacterial activity, and collagen synthesis and cell viability outcomes of adipocytes upon exposure to Abeliophyllum distichum Nakai (AdN). Antibacterial activity was measured through the Disc diffusion method to compare the growth ability of pathogenic microorganisms (E.coli, Salmonella). The absorbance was measured at 560 nm to calculate the active oxygen scavenging ability. Fibroblasts were dispensed in a 96-well plate at a density of 1 × 105 cells·well-1. The amount of procollagen was measured in each case using a procollagen type 1 C-peptide EIA KIT. The cytotoxicity of the Abeliophyllum distichum Nakai extract against animal adipocytes (Hanwoo backfat cells) was determined using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, a method that measures the conversion of MTS to Formazan by means of mitochondrial dehydrogenases. The concentrations of the samples were made to be 0.0125, 0025, 0.05, 0.1, and 0.5% and all were -completely absorbed into the disc in an incubator at 37℃ for 24 to 36 hours. For the 0.125 mg·disc-1, effects of Abeliophyllum distichum Nakai on the antioxidant effect, antibacterial activity, and cell viability of adipocytes were found. However, Abeliophyllum distichum Nakai had no effect on collagen synthesis, thus suggesting that AdN extracts may be useful for the prevention and/or treatment of obesity.

Antibacterial and phytochemical properties of Aphanamixis polystachya essential oil

  • Rahman, Md. Shahedur;Ahad, Abir;Saha, Subbroto Kumar;Hong, Jongki;Kim, Ki-Hyun
    • 분석과학
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    • 제30권3호
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    • pp.113-121
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    • 2017
  • Now a day's rise of new antibiotic resistant bacterial strains is a global threat. Ethnic people of India have been employing Aphanamixis polystachya (Wall.) R. Parker wood extract in healing cancerous wounds. The aim of this study was to evaluate the antimicrobial activity and to identify the medicinally potent chemicals in the essential oil extract of A. polystachya. The antibacterial properties of various organic extracts were evaluated against a range of bacteria (gram-positive and gram-negative bacteria) based on the disc diffusion method and GC-MS based analysis for finding active oil extract components. All extracts of A. polystachya leaves showed potential antibacterial activity, notably ethyl acetate, while petroleum ether extracts revealed highly sensitive activity against all tested bacteria (zones of inhibition ranging from 8.83 to 11.23 mm). In addition, the petroleum ether extract had the lowest MIC value (32 to $256{\mu}g/mL$) against E. coli, S. lutea, X. campestris, and B. subtilis bacteria. The major compounds detected in oil [${\beta}$-elemene (16.04 %), ${\beta}$-eudesmol (12.78 %), ${\beta}$-caryophyllene (19.37 %), ${\beta}$-selinene (11.32 %), elemol (5.76 %), and ${\alpha}$-humulene (5.68 %)] are expected to be responsible for the potent antimicrobial activity. The results of this study offer valuable insights into the potent role of A. polystachya essential oil extract in pharmaceutical and antibiotic research.

잘피(Zostera marina) 추출물의 항균효과에 대한 연구 (A Study of Antimicrobial Effect of Zostera marina Extracts)

  • 이소연;김보애;신동철;박관순;양재찬
    • 한국응용과학기술학회지
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    • 제33권2호
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    • pp.225-231
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    • 2016
  • 본 연구는 잘피 추출물의 화장품 소재로서 활용가능성을 평가하기 위해 건조된 잘피 전체, 뿌리, 잎 줄기를 70% 에탄올로 추출하여 항균활성을 확인하였다. 잘피 추출물의 항균활성은 Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Propionibacterium acnes 균주로 disc diffusion 방법을 통해 생육저해환(clear zone)을 측정하였다. 그 결과 잘피 뿌리 추출물처리군에서 Staphylococcus epidermidis의 clear zone이 $13.00{\pm}0.50mm$로 확인되었고, Staphylococcus aureus, Propionibacterium acnes, Pseudomonas aeruginosa의 clear zone은 각각 $11.75{\pm}0.25mm$, $12.00{\pm}0.50mm$, $12.25{\pm}0.25mm$로 확인되어 잘피 추출물의 항균 효능을 확인할 수 있었다.

서울동물원 야생동물의 임상 검체 내 Clostridium 균의 항생제 내성 분석 (Analysis of the anti-microbial susceptibility of Clostridium isolated on clinical specimens from captive wild animals in Seoul Zoo)

  • 이하늬;여용구;안상진;김종택
    • 한국동물위생학회지
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    • 제43권1호
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    • pp.31-37
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    • 2020
  • Clostridial bacteria are zoonotic agents, which cause severe necrotizing enteritis, pseudo-membrane colitis, enterotoxemia to both humans and animals. The objective of this study was to monitor the antibiotic resistance of Clostridium isolates on clinical specimens from wild animals in Seoul zoo for 5 years. Clostridium isolates were verified by using Vitek2 compact machine. Antibiotic susceptibility was assessed by antibiotic disc diffusion test, which was followed by Kirby-Bauer disc diffusion test method. The frequency of Antimicrobial resistance of Clostridium isolate was the greatest in gentamicin (87%), then in order of amikacin (80%). There were 55.6% of Clostridium isolates showed multiple drug resistance (MDR). These results showed that a lot of Clostridial bacteria from wild animals in Seoul zoo were acquired antibiotic resistance. Because of the wild animal's aggressive manner, it has been hard to collect clinical samples from wild animals in a zoo to exam antibiotic susceptibility. For these reasons, empirical use of antibiotics has been performed in frequently. It may cause to increase the emergence of antibiotic resistance bacteria. In addition, the antibiotic resistance bacteria from zoo animals can be spread to other wild animals which inhabit around the zoo. Therefore, regular monitoring of antibiotic resistance Clostridial bacteria is important to protect animals and humans from Clostridial diseases.

구강세균에 대한 구름버섯 추출물의 항균효과 (Antimicrobial activities of various extracts of Coriolus versicolor against oral bacteria)

  • 김별이;임근옥;한소라;김기화;오태진
    • 한국치위생학회지
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    • 제17권1호
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    • pp.111-122
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    • 2017
  • Objectives: Coriolus versicolor is an edible mushroom with physiological activities that has been used in traditional medicine. The aim of this study was to investigate the antimicrobial activity of extracts obtained from Coriolus versicolor against oral pathogens. Methods: The antimicrobial activities of various extracts of Coriolus versicolor were examined by disc diffusion assay, and minimum inhibitory concentration (MIC) of these extracts were also was determined by broth dilution method. The growth inhibition effect of extracts was measured at 600 nm for 12 hrs against Streptococcus ratti, Streptococcus criceti, Aggregati--bacter actinomycetemcomitans, Actinomyces viscosus, and Actinomyces israelii. Results: Coriolus versicolor extracts showed antimicrobial activities against all nine oral pathogens through disc diffusion assay. The ethanol extract and ethyl acetate extract differed significantly compared with acetone extract against Streptococcus ratti, Streptococcus criceti, Actinomyces viscosus, Actinomyces israelii and Aggregatibacter actinomycetemcomitans (p<0.05). These extracts exhibited MIC ranges of 2.63 to >10.50 mg/ml against the tested bacteria. The ethanol extract from Coriolus versicolor showed lower MIC values of 2.63 to 5.25 mg/ml. According to the obtained growth curve, the extracts of Coriolus versicolor were more effective against Actinomyces viscosus. Conclusions: The acetone, ethanol, and ethyl acetate extracts from Coriolus versicolor showed antimicrobial activities against Streptococcus mutans, Staphylococcus aureus, Streptococcus sanguinis, Streptococcus sobrinus, Streptococcus ratti, Streptococcus criceti, Aggregatibacter actinomycetemcomitans, Actinomyces viscosus, and Actinomyces israeli.i Therefore, they could be considered as natural oral antimicrobial agents against oral pathogens.