• Title/Summary/Keyword: Agar dilution method

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Standardization of an enzyme-linked immunosorbent assay for detection of antibody to avian reticuloendotheliosis virus (세망내피증 바이러스 항체검출을 위한 ELISA 표준화)

  • Sung, Haan Woo;Lee, Su Jeong
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.569-574
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    • 2005
  • Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

Degradation of Polyvinyl Alcohol in Dye-Processing Wastewater by Agar-Acrylamide Microbial Immobilization Method (한천-아크릴아마이드 미생물 고정화법에 의한 폐수 중 폴리비닐알콜의 분해)

  • 김재훈;김정목조무환
    • KSBB Journal
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    • v.10 no.3
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    • pp.241-248
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    • 1995
  • For the treatment of poorly biodegradable polyvinyl alcohol(PVA) in dye-processing wastewater, immobilized microbial beads were prepared by uslng agar-acrylamide method. PVA removal efficiency for the synthetic wastewater was 85% at the PVA volume loading rate of $3.1g/\ell$.day. In case of real desizing wastewater, PVA removal efficiency was 81.3% at the PVA volume loading rate of $3.25g/\ell$.day. In observation of cross section of immobilized bead passed 5 months with diameter of 2.4mm, the growth of cell was limited by the resistance of substrate and oxygen transfer for the inners region of more than 48% of bead radius from the surface. It was estimated that 70% of total removed PVA was degraded by the immobilized cells in the continuous immobilized reactor. Substrate utilization rate in the suspended reactor was decreased with increasing dilution rates above 0.083 hr-1, but that in the immobilized reactor was increased with increasing dilution rates up to 0.125hr-1. The substrate removal efficiency of immobilized reactor was much superior to that of suspended reactor with increasing dilution rates. Saturation constant of substrate utilization rate equation, Ks was $6.6 g PVA/\ell$, and maximum specific substrate utilization. k was 0.175g PVA/g cell.hr

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Isolation of Citrate-Utilizing Variants of Escherichia coli from Cattle, Pigs, Chickens, Pigeons and Wild Animals (동물유래(動物由來)의 Citrate이용대장균(利用大腸菌) 변이주(變異株)에 관하여)

  • Lee, Hun-Jun;Choi, Won-Pil
    • Korean Journal of Veterinary Research
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    • v.23 no.2
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    • pp.173-178
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    • 1983
  • This paper deals with the isolation of citrate-utilizing variants of Escherichia coli ($Cit^+$ E. coli) from the animals, their biochemical reactivity and antibiotic susceptibility, and whether the citrate utilizing ability is transmissible. One hundred arid twenty-three isolates of $Cit^+$ E. coli were obtained from cattle and pigs. but from other animals, no isolates were obtaied. Antibiotic susceptibility testing of $Cit^+$ E. coli was performed by the agar dilution method, using the following 9 antibiotics, chloramphenicol(CP), tetracycline(TC), streptomycine(SM), kanamycin(KM), colistin(CL), gentamicin(GM), cephaloridine(CR), aminobenzycillin and nalidixic acid(NA). All the variants tested were susceptible to KM, CL, GM, CR and NA. Of all the variants, 80(65%) were resistant to the drugs tested and resistance to TC and SM was most frequent. The transmission of the ability to utilize citrate on Simmons citrate agar at $37^{\circ}C$ was demonstrated in 78(67.8%) out of the 115 $Cit^+$ E. coli. There were no significant difference in the transfer rates of citrate utilizing ability between resistant and susceptible variants to above 9 drugs. Of 123 isolates, 8 were lost their citrate utilizing ability, spontaneously.

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Comparative Antimicrobial Susceptibility by the Methods of Test (검사방법(檢査方法)에 따르는 항균제감수성(抗菌劑感受性)의 비교(比較))

  • Park, Dong-Choon;Chun, Do-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.13 no.1
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    • pp.43-48
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    • 1978
  • The difference in antimicrobial susceptibility of multiply drug-resistant Salmonella typhi by the methods of test was tested against chloramphenicol(Cm), tetracycline(Tc), ampicillin(Ap), kanamycin(Km), and rifampicin(Rif), and the results were compared by the minimum inhibitory concentration(MIC). No appreciable difference was noted between MICs of Cm, Tc, and Rif measured by agar plate dilution(plate) method and broth dilution(broth) method. However, MICs of Km to about a half of test strains were 4 to 8-fold higher by broth method than plate method, and MICs of Ap by broth method were also a little higher than plate method in some strains. Cm and Rif were bacteriostatic rather than bactericidal to the most of test strains in high concentrations. Tc, Ap and Km were exclusively bactericidal to the test strains.

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The Change of Resistance of Phytophthora infestans to Metalaxyl and the Relationship with the Pathogenicity on Pepper Plants (국내 고추 역병균의 metalaxyl 저항성 변화 및 metalaxyl 저항성과 고추에 대한 병원성과의 상관 관계)

  • Yeon, Cho-Long;Lee, Soo-Min;Kim, Sun-Bo;Min, Gi-Young;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.12 no.3
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    • pp.270-276
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    • 2008
  • Through the agar dilution method on V-8 juice agar, sensitivity of Phytophthora capsici causing pepper Phytophthora blight to metalaxyl was investigated by using isolates obtained from infected pepper plants during 3 years from 2005 to 2007. By the lapse of time, $EC_{50}$ value to metalaxyl was decreased, showing 1.45, 0.83, and $0.32{\mu}g\;mL^{-1}$ in 2005, 2006, and 2007. None of 2007 isolates was found to be resistant to metalaxyl. Compared the sensitivity of P. capsici isolates to metalaxyl with those to mandipropamid and dimethomorph, there is not a cross resistance response between metalaxyl and mandipropamid/dimethomorph. The resistance to metalaxyl in pepper Phytophthora blight pathogen was not related with the mycelial growth on V-8 agar medium and the pathogenicity on pepper plants.

Anti-microbial Activity of Platycodon Grandiflorum Extracts Against Oral Microbes (도라지 추출물의 구강미생물에 대한 항균효과)

  • Jung, So-Young;Lee, Cheon-Hee;Ahn, Sun-Ha
    • The Korean Journal of Health Service Management
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    • v.13 no.2
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    • pp.135-142
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    • 2019
  • Objectives: The objective of this study was to assess the antimicrobial effect of Platycodon grandiflorum extracts against oral microorganisms. Methods: The anti-microbial activity and minimal inhibitory concentration were measured the agar dilution method. Results: Platycodon grandiflorum extracts grew in the free agar plates all of the oral microorganisms. In the bark-free Platycodon grandiflorum extracts all the oral microorganisms grew in the free agar plates. Growth was inhibited at a concentration of 0.5 mg/ml. Oral microorganisms showed an absence of growth at a concentration of 1 mg/ml. Conclusions: It was confirmed that the extracts of Platycodon grandiflorum having a higher saponin content than the bark - free Platycodon grandiflorum extract showed excellent antimicrobial effect.

Selection of Fungicides for the Control of Soybean Black Root Rot Caused by Calonectria ilicicola (콩 검은뿌리썩음병 방제를 위한 살균제 선발)

  • Park, Seong-Woo;Kang, Beom-Kwan;Kim, Hong-Sik;Woo, Sun-Hee;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.11 no.1
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    • pp.18-26
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    • 2007
  • Fungicidal screening was performed to control soybean black root rot caused by Calonectria ilicicola through in vitro and greenhouse assays. In in vitro assay, 25 fungicides were assessed by an agar dilution method and a 96-well microtiter plate method. While protective fungicides including dithianon, dichlofluanid, mancozeb, and captan showed a very low activity against the mycelial growth C. ilicicola SC03-15 in the agar dilution method, they displayed potent inhibitory activity against spore germination in a 96-well microtiter plate method with $EC_{50}$ values of 4.65, 0.61, 4.64, and $0.29{\mu}g\;mL^{-1}$, respectively. Ergosterol biosynthesis-inhibiting (EBI) fungicides showed different antifungal activity against mycelial growth and spore germination according to molecules. Difenconazole displayed higher antifungal activity against spore germination rather than mycelial growth, and prochloraz inhibited potently both mycelial growth and spore germination with EC50 values less than $1.8{\mu}g\;ml^{-1}$. In contrast, the other EBI fungicides inhibited more highly mycelial growth than spore germination. Carbendazim+diethofencarb and dazomet also inhibited both mycelial growth and spore germination of C. ilicicola SC03-15 at very low concentrations. In greenhouse assay, carbendazim+diethofencarb effectively controlled a soybean black root rot by drenching 2 days before or after inoculation. In addition, tebuconazole showed potent curative activity against soybean black root rot.

Synthesis and $\beta$-lactamase inhibitory activity of 6-exomethylene penamsulfone derivatives-III (In vitro $\beta$-lactamase inhibitory activity and comparative susceptibility test of 6-exomethylene penamsulfone derivatives)

  • Park, Hee-Suk;Yoon, Sang-Bae;Chaeuk Im;Yim, Chul-Bu
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.72-72
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    • 1997
  • 합성한 5종의 6-Exomethylene penamsulfone 유도체의 Type I, Type II, Type III, TypeIV, TEM 효소에 대한 $\beta$-lactamase저해효과를 spectrophotometric assay방법으로 측정하였다. 또한 여기서 우수한 효소억제활성을 보여준 3종의 화합물을 가지고 In vitro antibacterial activity를 Agar dilution method법으로 27종의 $\beta$-lactamase생성균주에 대하여, Sulbactam, Ampicillin 및 Cefoperazone과 병용투여하여 MIC를 측정하였다.

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In Vitro Activity of Cyclic Dipeptides Against Gram-Positive and Gram-Negative Anaerobic Bacteria and Radioprotective Effect on Lung Cells

  • RHEE KI-HYEONG
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.158-162
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    • 2006
  • Cyclic dipeptides isolated from Streptomyces sp. have been shown to have antimicrobial activity as well as other potentially useful biological activities. The purpose of this study was to compare the in vitro activity of two cyclic dipeptides combined against anaerobic bacteria with the activity of other antimicrobial agents. Specifically, the in vitro activity of the combination of two cyclic dipeptides was investigated against 140 clinical isolates of anaerobic bacteria by the agar dilution method and was compared with that of erythromycin, cefoxitin, imipenem, clindamycin, and metronidazole. The cyclic dipeptide combination and imipenem were the most active antimicrobial agents tested. In addition, the cyclic dipeptide combination had a radioprotective effect on five normal human lung fibroblast cells, showing survival rates higher $(>90\%)$ than either of the two cyclic dipeptides alone $(<80\%)$.

Acute Toxicity and Antimicrobial Activity of 1-Deoxynojirimycin (1-Deoxynojirimycin의 급성독성 및 항균효과)

  • 백남수;김영만
    • The Korean Journal of Food And Nutrition
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    • v.11 no.6
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    • pp.629-634
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    • 1998
  • 1-Deoxynojirimycin which is a potent intestinal ${\alpha}$-glucosidase inhibitor was purified from the culture broth by ion exchange chromatography, Sephadex LH20 column chromatography, TSK gel chromatography and HPLC respectively. Acute toxicity of 1-deoxynojirimycin, which was loaded through the oral as dose of 200mg/kg, was investigated in IRC mouse. None of the tested IRC mice were not dead and increase of body weight showed also the same results in comparison with control mice. The antimicrobial susceptibility of 20 pathogenic strains against 3 antidiabetic compounds (1-deoxynojirimycin, AO-128, acarbose) were obtained by agar dilution method. All of the three antidiabetic compounds has very weak antimicrobial activity (MIC>100$\mu\textrm{g}$/ml).

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