• Title/Summary/Keyword: Agar diffusion method

Search Result 171, Processing Time 0.036 seconds

Development of an Agar Diffusion Method to Measure Elastase Inhibition Activity Using Elastin-Congo Red

  • Jung Kyung-Hwan;Kim Hyun-Joo
    • Journal of Microbiology and Biotechnology
    • /
    • v.16 no.8
    • /
    • pp.1320-1324
    • /
    • 2006
  • The pancreatic and neutrophil elastases are associated with several illnesses including lung and vascular diseases, various cancers, and pancreatitis. The development of a potent and specific inhibitor to the elastases could lead to new therapies. In this study, an agar diffusion method was modified to include a substrate-dye conjugate (Elastin-Congo red) as a substrate of elastase and an indicator of elastase inhibitory activity. The Elastin-Congo red agar plates consisted of 0.1 % Elastin-Congo red and 2.5% agar. The elastase and elastase inhibitors were simultaneously loaded into wells, ultimately resulting in halo formations in which the halo diameter decreased as the concentration of elastase inhibitor increased. The concentration of elastase inhibitor in the samples, therefore, was inversely proportional to the halo diameters. This simplified method provided an excellent correlation with the standard microplate technique, which uses a chromogenic substrate. The concentration of elastase inhibitor obtained from the culture supernatant of a recombinant elastase inhibitor produced by the yeast Pichia pastoris was easily determined. This study has established a simple modified and inexpensive agar diffusion method that is potentially useful for the identification, quantification, and screening of new elastase inhibitors.

Determination of Vitamin $B_12$ by Agar Diffusion Method (원통평판법을 이용한 비타민 $B_12$의 정량)

  • Lee, Sung-Ho;Cho, Chin-Sung;Song, Young-Joon
    • Journal of Pharmaceutical Investigation
    • /
    • v.21 no.2
    • /
    • pp.79-84
    • /
    • 1991
  • The agar diffustion method using Escherichia coli was investigated for determination of Vitamin $B_{l2}$ instead of turbidimetric method using Lactobacillus leichannii (USP XXII method). The turbidimetric method is difficult to control the test organism and it has complicated procedure. From this study, it was found that the agar diffusion method on the determination of Vitamin $B_{12}$ in pharamaceutical preparation is simple and convenient as compared with turbidimetric method. Also we found that the coefficient of variation in reproducibility and the standard error in recovery were 2.18% and 1.83%, respectively.

  • PDF

Diffused Quantity Prediction Model and Diffusion Coefficient of Gel Food in the Mass Transfer Process (겔 식품의 물질전달공정 중 확산량 예측모델 및 확산계수)

  • Lee, Won-Young;Choi, Yong-Hee
    • Korean Journal of Food Science and Technology
    • /
    • v.23 no.2
    • /
    • pp.217-223
    • /
    • 1991
  • A capillary method was used to evaluate the properties of mass transfer process and diffusion coefficients in the gel food. Amaranth dye was selected as a diffusant material to visualize the degree of diffusion procedure easily. After contacting cylinder containing agar gel with amaramth dye solution for some hours, the gel was cut to five segments by 0.5 cm in length. The diffusant concentration from the segments were measured by the spectrophotometer at 523 nm. Prediction models for the diffused quantities in gel food were established by the regression program of SPSS package program. Generally, diffusion coefficient can be calculated by Fick's second law, however, it will be determined by using numerical analysis method more easily. Finally the diffusion coefficients in this research were calculated by arithmetic mean of the measured values. As raising gel agent concentration, the mean diffusion coefficient tended to decrease because the obstruction effect came to become significant.

  • PDF

Study on Antimicrobial Activity of Extracts from Fritillaria unibracteata Hsiao et K.C. Hsia and F. ussuriensis Maxim.

  • Moon, Jung-Wook;Chen-Zhi, Chen-Zhi;Song, Won-Seob;Baek, Seung-Hwa
    • Korean Journal of Plant Resources
    • /
    • v.24 no.6
    • /
    • pp.719-723
    • /
    • 2011
  • Antimicrobial activities of methanol, ethanol, water, and $CH_2Cl_2$ extracts from Fritillaria unibracteata Hsiao et K.C. Hsia and F. ussuriensis Maxim. were investigated by disk-agar diffusion method. The result showed comparatively strong antimicrobial activity against several microorganisms. The extracts from F. unibracteata and F. ussuriensis dosedependently increased the activity. However, water and $CH_2Cl_2$ extracts showed no antimicrobial activity against 7 microorganisms. Especially, against the most sensitive microorganism Staphylococcus epidermidis, methanol extracts at highest concentration of 20 mg/mL exhibited the largest clear zone on plate by 6-12 mm and ethanol extracts on plate by 6-10 mm.

Investigation of Antimicrobial Activity of Brown Algae Extracts and the Thermal and pH Effects on Their Activity

  • Lee, So-Young;Kim, Jin-Hee;Song, Eu-Jin;Kim, Koth-Bong-Woo-Ri;Hong, Yong-Ki;Lim, Sung-Mee;Ahn, Dong-Hyun
    • Food Science and Biotechnology
    • /
    • v.18 no.2
    • /
    • pp.506-512
    • /
    • 2009
  • The antimicrobial activity of water and ethanol extracts from 30 species of algae was measured using the agar diffusion method and minimum inhibitory concentration (MIC) test. In agar diffusion method, the 95% ethanol extracts from 12 of the algae showed growth inhibition against the tested microorganisms. In particular, Ishige okamurai, Ecklonia stolonifera, Sargassum siliquastrum, Sargassum thunbergii, Colpomenia bullosa, and Ecklonia cava had strong antibacterial activities against Gram-positive bacteria at 4 mg/mL. In the results of the MIC test, S. siliquastrum showed the most antimicrobial activity, where its MIC values ranged from 0.005 to 0.0075% against Listeria monocytogenes, Clostridium perfringens, and Basillus subtilis. In the thermal stability test, for the ethanol extracts of I. okamurai, E. cava, S. siliquastrum, S. thunbergii, and C. bullosa, the extracts proved to maintain high antimicrobial activities when they were treated at $121^{\circ}C$ for 15 min. In the pH stability test, the antimicrobial activity of the S. siliquastrum ethanol extract was stable from pH 2 to 10, whereas the activity of the other species ethanol extracts were weakened under pH 10 against several microbes.

In situ isolation and characterization of the biosurfactants of B. Subtilis

  • Akthar, Wasim S.;Aadham, Mohamed Sheik;Nisha, Arif S.
    • Advances in environmental research
    • /
    • v.9 no.3
    • /
    • pp.215-232
    • /
    • 2020
  • Crude oils are essential source of energy. It is majorly found in geographical locations beneath the earth's surface and crude oil is the main factor for the economic developments in the world. Natural crude oil contains unrefined petroleum composed of hydrocarbons of various molecular weights and it contains other organic materials like aromatic compounds, sulphur compounds, and many other organic compounds. These hydrocarbons are rapidly getting degraded by biosurfactant producing microorganisms. The present study deals with the isolation, purification, and characterization of biosurfactant producing microorganism from oil-contaminated soil. The ability of the microorganism producing biosurfactant was investigated by well diffusion method, drop collapse test, emulsification test, oil displacement activity, and blue agar plate method. The isolate obtained from the oil contaminated soil was identified as Bacillus subtilis. The identification was done by microscopic examinations and further characterization was done by Biochemical tests and 16SrRNA gene sequencing. Purification of the biosurfactant was performed by simple liquid-liquid extraction, and characterization of extracted biosurfactants was done using Fourier transform infrared spectroscopy (FTIR). The degradation of crude oil upon treatment with the partially purified biosurfactant was analyzed by FTIR spectroscopy and Gas-chromatography mass spectroscopy (GC-MS).

Antimicrobial Activity of Methyl Gallate isolated from the Leaves of Glochidion superbum Against Hospital Isolates of Methicillin Resistant Staphylococcus aureus

  • Ahmed, Mohammed Dahiru;Taher, Muhammad;Maimusa, Alhaji Hamusu;Rezali, Mohamad Fazlin;Mahmud, Mohammed Imad Al-deen Mustafa
    • Natural Product Sciences
    • /
    • v.23 no.1
    • /
    • pp.5-8
    • /
    • 2017
  • An antimicrobial compound has been isolated from the leaves of Glochidion superbum. The compound was determined as methyl 3, 4, 5-trihydroxybenzoate (methyl gallate), based on ultraviolet (UV), infrared (IR), nuclear magnetic resonance (NMR) and mass spectroscopy (MS) analysis. The isolated compound exhibited potent antimicrobial activity against three clinical isolates of methicillin resistant Staphylococcus aureus (MRSA) by qualitative agar disc diffusion method and quantitative broth dilution method. Agar disc diffusion was done in a dose-dependent manner for each bacterial isolate at disc potencies of 25, 50, 100, and $150{\mu}g/disc$. The zones of inhibition were on average equal to 12.27, 14.20, 15.43, and 24.17 mm respectively. The inhibition zones were compared with that of vancomycin disc at $30{\mu}g$ as a reference standard. The MIC and MBC values were $50{\mu}g/ml$ and $100{\mu}g/ml$ respectively. The results of anti MRSA activity were analyzed using one-way ANOVA with Turkey's HSD and Duncan test. In conclusion, methyl gallate which was isolated from G. superbum showed the inhibition activity against methicillin resistant S. aureus.

Antibacterial effect of self-etching adhesive systems on Streptococcus mutans

  • Kim, Seung-Ryong;Shin, Dong-Hoon
    • Restorative Dentistry and Endodontics
    • /
    • v.39 no.1
    • /
    • pp.32-38
    • /
    • 2014
  • Objectives: In this study, we evaluated the antibacterial activity of self-etching adhesive systems against Streptococcus mutans using the agar diffusion method. Materials and Methods: Three 2-step systems, Clearfil SE Bond (SE, Kuraray), Contax (CT, DMG), and Unifil Bond (UnB, GC), and three 1-step systems, Easy Bond (EB, 3M ESPE), U-Bond (UB, Vericom), and All Bond SE (AB, BISCO) were used. 0.12% chlorhexidine (CHX, Bukwang) and 37% phosphoric acid gel (PA, Vericom) were used as positive controls. Results: The antibacterial activity of CHX and PA was stronger than that of the other groups, except SE. After light activation, the inhibition zone was reduced in the case of all 2-step systems except CT. However, all 1-step systems did not exhibit any inhibition zone upon the light activation. Conclusions: SE may be better than CT or UnB among the 2-step systems with respect to antibacterial activity, however, 1-step systems do not exhibit any antibacterial activity after light curing.

Errors of Antibiotic Susceptibility Testing from Automated and Manual Systems in Clinical Isolates of Acinetobacter baumannii

  • Sung, Ji Youn;Oh, Ji-Eun;Kim, Eun Sun
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.45 no.1
    • /
    • pp.21-25
    • /
    • 2013
  • Acinetobacter baumannii is an aerobic, gram-negative and glucose-non-fermenting bacterium, which has emerged as a serious opportunistic pathogen. Many clinical microbiology laboratories use the Vitek 2 system for the routine antimicrobial susceptibility testing process, including testing on A. baumannii isolates. However, in case of amikacin, it is now recommended to perform additional antimicrobial susceptibility testing for A. baumannii strains due to the relatively lower minimum inhibitory concentration (MIC) in the Vitek 2 system compared to conventional reference methods. In our study, we assessed MIC for amikacin susceptibility testing of A. baumannii isolates in the Vitek 2 system, the agar dilution, Etest, and disk diffusion method. We collected 40 gentamicin-resistant, A. baumannii strains (amikacin MIC by Vitek 2:${\leq}2{\mu}g/mL$, 2 isolates; $4{\mu}g/mL$, 34 isolates; $8{\mu}g/mL$, 4 isolates) from a University hospital and compared the Vitek 2 system to other reference methods for testing susceptibility to amikacin. The Vitek 2 system showed major errors in all of the 40 isolates, yielding a low MIC. The results of our study strongly suggested that the Vitek 2 system was not a reliable method to test the MICs of gentamicin; ranging from ${\geq}16{\mu}g/mL$ for amikacin susceptibility. Other tests, such as agar dilution, Etest, or disk diffusion methods, should be paralleled to determine the MIC of amikacin in A. baumannii.

  • PDF

Antimicrobial Activity of Essential Oils of Curry Spices (카레 향신료 정유성분의 항균성)

  • Chung, Chang-Ki;Park, Oan-Kyu;Yoo, Ik-Je;Park, Ki-Moon;Choi, Chun-Un
    • Korean Journal of Food Science and Technology
    • /
    • v.22 no.6
    • /
    • pp.716-719
    • /
    • 1990
  • The essential oils were isolated by steam distillation from 13 spices used for curry. Antimicrobial activity of essential oils for two strains of Gram(+) bacteria, Gram(-) bacteria, lactic acid bacteria, yeast and mold were investigated by agar diffusion method. 5 spice essential oils(clove, cumin, nutmeg, oregano, rosemary) having high antimicrobial activity were selected and their minimal inhibitory concentration(MIC) were measured. Very low concentration ($0.2{\sim}9\;mg/ml$) of 5 spice essential oils were sufficient to prevent microbial growth. The data show that Gram(+) bacteria were more sensitive to the antimicrobial compounds in spices than Gram(-). But though Gram(+) bacteria, lactic acid bacteria were less sensitive to the compounds than Gram(-).

  • PDF