• Title/Summary/Keyword: disc diffusion test

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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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    • v.63 no.1
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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.

Comparison of Harboring the Resistance Gene and Disc Diffusion Susceptibility Test Result in Staphylococcus pseudintermedius from the Bacterial Dermatitis (세균성 피부염 개에서 분리된 Staphylococcus pseudintermedius에서 항생제 감수성 검사와 내성 유전자 획득의 비교)

  • Jang, Hye-Jin;Son, Hyoung-Won;Kang, Hyo-Min;Han, Jae-Ik;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.158-161
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    • 2015
  • Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.

Antimicrobial Effect of Kaempferol on Psychrotrophic Bacillus cereus Strains Outbreakable in Dairy Products

  • Lee, Kyoung-Ah;Moon, Sun-Hee;Kim, Kee-Tae;Nah, Seung-Yeol;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.31 no.2
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    • pp.311-315
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    • 2011
  • The objective of this study was to evaluate the antimicrobial effects of various natural flavonoids against growth of psychotropic Bacillus cereus strains, which cause dairy food outbreaks. Flavonoids were first screened for their ability to inhibit growth of B. cereus strains using the paper-disc diffusion test. Second, the growth inhibitory effect of selected flavonoids was evaluated in tryptic soy broth supplemented with 0.6% yeast extract, and the bactericidal effect of the flavonoids was measured in 0.8% (w/v) NaCl solution. Based on the paper-disc diffusion test, kaempferol was effectively active against B. cereus P14 and B. cereus KCCM 40935. Kaempferol had an antimicrobial effect at concentrations greater than 100 ${\mu}M$, and the numbers of B. cereus P14 and B. cereus KCCM 40935 decreased by 3.55 and 1.5 log cycles, respectively. The cell numbers of B. cereus P14 and B. cereus KCCM 40935 treated with 50 ${\mu}M$ kaempferol were reduced by 4.18 and 2.84 log cycles during a 24 h incubation to test the bactericidal effect of kaempferol (p<0.05). The results indicate that kaempferol had the greatest antimicrobial effect among the psychotropic B. cereus strains and the natural flavonoids tested.

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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    • v.21 no.3
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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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Antimicrobial Activity of the Ethanol Extract from Rubus coreanum against Microorganisms Related with Foodborne Illness (복분자 에탄올 추출물의 식중독 관련 위해 세균에 대한 항균활성 분석)

  • Jeon, Yeon-Hee;Sun, Xiaoqing;Kim, Mee-Ra
    • Korean journal of food and cookery science
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    • v.28 no.1
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    • pp.9-15
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    • 2012
  • This study analyzed the antibacterial activity of a Rubus coreanum (Bokbunja) ethanol extract. The antimicrobial activity was determined by disc diffusion, minimum inhibitory concentration (MIC), and growth inhibition methods with seven kinds of bacteria related to foodborne illness (Bacillus cereus, Escherichia coli, Escherichia coli O157:H7, Listeria monocytogenes, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhimurium). In the results, disc diffusions of the ethanol extract from R. coreanum (9.8-17.5 mm at $4,000{\mu}g/disc$) clearly showed the antimicrobial activity of the extract against all tested microorganisms. Rubus coreanum promoted an inhibitory effect as follows: E. coli O157:H7 > P. aeruginosa > L. monocytogenes > E. coli > S. aureus > B. cereus ${\geq}$ S. typhimurium. In the MIC test, R. coreanum showed high antimicrobial effect against L. monocytogenes at 500 ppm. Moreover, the R. coreanum ethanol extract showed strong growth inhibition against microorganisms, similar to the MIC results. These results show that a R. coreanum ethanol extract has powerful antimicrobial activity against all tested microorganisms, suggesting that R. coreanum will be useful as a potential natural preservative.

Growth-inhibiting Effects of Brazilian and Oriental Medicinal Plants on Human Intestinal Bacteria

  • Kim, Moo-Key;Lee, Sung-Eun;Lee, Hoi-Seon
    • Journal of Applied Biological Chemistry
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    • v.43 no.1
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    • pp.54-58
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    • 2000
  • Methanol extracts of 27 Brazilian plant samples and 10 oriental medicinal plant samples (27 families), using spectrophotometric and paper disc agar diffusion methods under anaerobic conditions, were tested in vitro for their growth-inhibiting activities against Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium adolescentis, Clostridium perfringens, and Bacteroides fragilis. The responses varied with bacterial strains, plant species, and tissues sampled. In a test with B. longum and B. bifidum(20 mg/disc), extracts of Acanthopanax sessilifolinus stem bark and Ampelozizyphus amazonicus leaves strongly inhibited the growth of B. longum, whereas other plant samples did not inhibit any intestinal bacteria tested. At 5 mg/disc, adding extracts of Aralia eleta, Euterpe oleracea, and Syzygium guineense to the media strongly inhibited the growth of C. perfringens and B. fragilis without growth inhibition of B. adolescentis, B. longum, and B. bifidum. Extracts of Jacaranda mimosifolia and Ulmus paraifolia significantly inhibited the growth of C. perfringens and B. fragilis as well as B. adolescentis. These results may be indications of at least one of the pharmacological actions of the five Brazilian plants but not oriental medicinal plants tested.

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Antimicrobial effects of curcumin against pathogenic bacteria in fish (어류의 병원성 세균에 대한 curcumin의 항균효과)

  • Heo, Gang-Joon;Kang, Jin-Hui;Shin, Gee-Wook
    • Korean Journal of Veterinary Service
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    • v.36 no.4
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    • pp.297-301
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    • 2013
  • The present study was to investigate anti-microbial effects of curcumin on major bacterial pathogens for farmed fish, such as Aeromonas hydrophila, A. salmonicida subsp. masoucida, A. salmonicida subsp. salmonicida, Edwardsiella tarda, Vibrio vulnificus, V. paraheamolyticus using disk diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) tests. In disc diffusion test, curcumin exhibited concentration-dependent antimicrobial activities to all bacteria pathogens used in the study. Antimicrobial effects of curcumin was found differently depending on bacterial species when determined by MIC or MBC tests. For examples, E. tarda and A. hydrophila was respectively the most sensitive bacterium for bacteriostatic and bacteriocidal effect of curcumin. Collectively, curcumin could be a potential natural drug for controlling pathogenic bacteria in the aquaculture industry.

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

  • Lee, Hany;Yeo, Yong-Gu;Ahn, Sangjin;Kim, Jong-Taek
    • Korean Journal of Veterinary Service
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    • v.43 no.1
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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.

New Azafluorenone Derivative and Antibacterial Activities of Alphonsea cylindrica Barks

  • Talip, Munirah Abdul;Azziz, Saripah Salbiah Syed Abdul;Wong, Chee Fah;Awang, Khalijah;Naz, Humera;Bakri, Yuhanis Mhd;Ahmad, Mohamad Syahrizal;Litaudon, Marc
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.151-156
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    • 2017
  • A phytochemical study of Alphonsea cylindrica King (unreported) has led to the isolation of six alkaloids. The compounds were identified as kinabaline (1; azafluorenone alkaloid), muniranine (2), O-methylmoschatoline (3; oxoaporphine alkaloid), lysicamine (4), atherospermidine (5) and N-methylouregidione (6; 4, 5-dioxoaporphine alkaloid). The structures of the isolated compounds were determined based on the spectroscopic techniques and by comparison with data reported in the literature. Alkaloid 2 was isolated as a new derivative of azafluorenone while alkaloids 1, 3 - 6 were isolated for the first time from Alphonsea species. In addition, alkaloid 3 and 4 showed inhibition zone against Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus cereus in disc diffusion test. The minimum inhibition concentration (MIC) values of lysicamine (4) against S. aureus, B. cereus and P. aeruginosa were found to be smaller than O-methylmoschatoline (3). Therefore, the reported antibacterial activity showed the potential of this plant as natural antibacterial agent and supported the documented traditional use of Alphonsea sp. in the treatment of diarrhea and fever.

The Trend of Antimicrobial Resistance of Escherichia coli isolated from Healthy Volunteers of Community and Hospital Patients in Incheon (지역사회 및 병원 임상검체에서 분리한 대장균의 항생제 내성 양상)

  • Kim, Yong-Hui;Go, Jong-Myeong;Gong, Young-Woo;Oh, Bo-Young;Kim, Jung-Hee;Kim, Hye-Young;Lee, Mi-Yeon;Koh, Yeon-Ja;Hwang, Kyoung-Wha;JeGal, Seung;Lee, Jae-Mann
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.252-256
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    • 2006
  • We monitored antibiotic resistance of Escherichia coli isolates from healthy volunteers of community and hospital patients from February to July in 2006. From disc diffusion test on 4915 E. coli isolates from healthy volunteers of the community, the resistance rates were as follows; tetracycline resistant, 46.6%; ampicillin resistant, 41.1%; ticarcillin resistant,37.9%. From disc diffusion test on 120 E. coli isolates from hospital patients, the resistance rates were as follows: ampicillin resistant, 66.9%; ticarcillin resistant, 63.8%; tetracycline resistant, 47.2%. Extended spectrum $\beta$-lactamase producing E. coli were isolated 0.6% and 4.1% from healthy volunteers and hospital patients.