• Title/Summary/Keyword: dilution assay

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Antibacterial activity of new quinolones against Edwardsiella tarda isolated from eel tanks (양만장에서 분리된 Edwardseilla tarda균에 대한 new quinolone계 약물의 항균작용)

  • Choi, Min-Soon;Kim, Kyong-Ho;Seo, Young-Ho;Choi, Sang-Hoon;Park, Kwan-Ha
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.185-193
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    • 1996
  • Extensive acquirement of drug resistance to traditional antibacterial agents poses a serious problem to eel aquaculturists. To collect the basic information for new drug development in the future, we assessed the in vitro antibacterial efficacy of 14 new quinolones with 75 isolates of Edwardsiella tarda from local aquaculture tanks of Anguilla japonica. Of all tested quinolones under development or marketed for human use, DU-6859 was most potent with its $MIC_{50}$ value of $0.05{\mu}g$/ml in broth microdilution assay. The drugs whose $MIC_{50}$ values ranged from 0.2 to $0.78{\mu}g$/ml were T-3762, Bay-y3118, ciprofloxacin, norfloxacin, ofloxcin and tosufloxacin. The weakest group of drugs, with their $MIC_{50}$ being 1.56-$3.13{\mu}g$/ml, were difloxacin, sparfloxacin, fleroxacin, Q-35, amifloxacin, lomefloxacin and enoxacin. The number of resistant strains, when arbitrarily defined with their MICs of $\geq6.25{\mu}g$/ml, was : 3 to T-3762, 3 to Bay-y3118, 44 to difloxacin, 16 to sparfloxacin, 13 to ciprofloxacin, 19 to fleroxacin, 36 to Q-35). 31 to amifloxacin, 5 to norfloxacin, 13 to ofloxacin, 31 to lomefloxacin, 41 to enoxacin, 12 to tosufloxacin and 0% to DU-6859, respectively. This information can be taken into consideration for the future development of fisheries antibacterial quinolones.

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Study on Soluble Concentrate Formulation and Quality Control Techniques of a Microbial Insecticide "Bt-Plus" (미생물살충제 "비티플러스" 액상 제형화 및 품질 분석 기술에 관한 연구)

  • Eom, Seonghyeon;Park, Hyeonji;Kim, Kyusoon;Hong, Youkyeong;Park, Jiyeong;Choi, Bongki;Kim, Joonsung;Kim, Kunwoo;Kang, Moonsoo;Yang, Kyunghyung;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.52 no.2
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    • pp.115-123
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    • 2013
  • A microbial insecticide "Bt-Plus" has been developed to enhance an insecticidal efficacy of an entomopathogenic bacterium, Bacillus thuringiensis (Bt). However, its wettable powder formulation is not preferred by farmers and industry producers due to relatively high cost. This study aimed to develop a soluble concentrate formulation of Bt-Plus. To this end, an optimal mixture ratio of two bacterial culture broths was determined to be 5:4 (v/v) of Bt and Xenorhabdus nematophila (Xn) along with 10% ethanol preservative. In addition, Bt broth was concentrated by 10 times to apply the mixture at 1,000 times fold dilution. The resulting liquid formulation was sprayed on cabbage crop field infested by late instar larvae of the diamondback moth, Plutella xylostella. The field assay showed about 77% control efficacy at 7 days after treatment, which was comparable to those of current commercial biopesticides targeting P. xylostella. For storage test in both low and room temperatures, the liquid formation showed a relatively stable control efficacy at least for a month. To develop a quality control technique to exhibit a stable control efficacy of Bt-Plus, Bt spore density ($5{\times}10^{11}$ spores/mL) and eight active component concentrations of Xn bacterial metabolites in the formulation products have been proposed in this study.

Fungicide Resistance of Fusarium fujikuroi Isolates Isolated In Korea (우리나라에서 분리한 벼 키다리병균(Fusarium fujikuroi)의 살균제 저항성)

  • Lee, Yong-Hwan;Kim, So-Yeon;Choi, Hyo-Won;Lee, Myeong-Ji;Ra, Dong-Soo;Kim, In-Seon;Park, Jin-Woo;Lee, Se-Weon
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.427-432
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    • 2010
  • Rice bakanae disease caused by Fusarium fujikuroi is one of the most serious rice diseases in Korea. From 2006 to 2009, 118 F. fujikuroi isolates were collected from various regions of rice fields in Korea. Resistance assay of 118 F. fujikuroi isolates to prochloraz, tebuconazole, and benomyl, were performed using agar dilution method. To investigate inhibitory effects of the fungicides, minimum inhibitory concentration of mycelial growth (MIC) and effective concentration inhibiting mycelial growth by 50% ($EC_{50}$) for 118 isolates were calculated using Sigmaplot 8.02 (Antro, SPSS UK, Ltd). Based on the means of $EC_{50}$ values, baseline resistance values were determined as $0.5{\mu}g{\cdot}mL^{-1}$ for prochloraz, $5.0{\mu}g{\cdot}mL^{-1}$ for tebuconazole and $2.5{\mu}g{\cdot}mL^{-1}$ for benomyl. Number of resistant isolates to each fungicide was 17, 19 and 43 for prochloraz, tebuconazole and benomyl, respectively. Furthermore, 4 isolates showed the double resistance to both prochloraz and tebuconazole, 6 isolates to prochloraz and benomyl, and 11 isolates to tebuconazole and benomyl. Isolates CF366 and LF335 isolated from Gyeongbuk province were resistant to the three fungicides tested, prochloraz, tebuconazole and benomyl.

Antimicrobial and Antioxidative Activities of the Extracts from Walnut (Juglans regia L.) Green Husk (호두과피 추출물의 항산화 및 항균활성)

  • Han, Kook-Il;Kim, Mi ran;Jo, Bu Kyung;Kim, Min Ji;Kang, Min Joo;Park, Ki-hyoun;Koo, Ye eun;Kim, Byeongseong;Jung, Eui-Gil;Han, Man-Deuk
    • Journal of Life Science
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    • v.25 no.4
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    • pp.433-440
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    • 2015
  • Several studies suggest that regular consumption of walnuts may have beneficial effects against oxidative stress-mediated disease such as cancer. The present study reports the total phenolic and flavonoid contents, together with the antioxidant and antibacterial activities of several solvent extracts (methanol, n-hexane, ethyl acetate, n-butanol, and water) obtained from walnut (Juglans regia L.) green husk. MIC (minimal inhibitory concentration) values of the walnut extracts for 8 human pathogenic bacteria strain were determined using agar dilution method. Antioxidant activity of extracts were assessed using DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid)) assays, EC50 of DPPH and ABTS scavenging activities, and determination of total phenolic and flavonoid content and its correlation with DPPH and ABTS scavenging capacities. Among the six extracts, ethyl acetate extract (EtOAc Ex) showed the highest antimicrobial activity at 3.2 mg/ml of MICs against Staphylococcus aureus SG511. Total flavonoids and polyphenol contents of EtOAc Ex were 42.48 mg of quercetin equivalents (QE)/g and 223.25 mg of gallic acid equivalents (GAE)/g respectively. The highest antioxidative potential was shown by the sample extracted with EtOAc Ex (EC50=13.43 μg/ml for DPPH and EC50=41.83 μg/ml for ABTS radical scavenging activity assay). These results showed that J. regia green husk extracts can be used as an easily accessible source of natural antibacterial agents and natural antioxidants.

Antibiotic Susceptibility of Bacteria Isolated from Infected Root Canals (감염근관에서 분리 배양한 세균의 수종 항생제에 대한 감수성 조사)

  • Lim, Sang-Soo;Kim, Mi-Kwang;Min, Jeong-Beom;Kim, Min-Jung;Park, Soon-Nang;Hwang, Ho-Keel;Kook, Joong-Ki
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.185-194
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    • 2006
  • The aim of this study was to identify the bacteria isolated from endodontic lesions by cell culture and to determine the antimicrobial susceptibility of them against 8 antibiotics. The necrotic pulpal tissues were collected from 27 infected root canals, which were diagnosed as endodontic infection. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing $500{\mu}l\;of\;1{\times}PBS$. The sample solution was briefly mixed and plated onto a BHI-agar plate containing 5% sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 2 to 5 days. The bacteria grown on the agar plates were identified by comparison of 16S rRNA gene (rDNA) sequencing method at the species level. To test the sensitivity of the bacteria isolated from the infected root canals against 8 antibiotics, minimum inhibitory concentrations (MIC) were determined using broth dilution assay. The data showed that 101 bacterial strains were isolated and were identified. Streptococcus spp. (29.7%) and Actinomyces spp. (21.8%) were predominantly isolated. The 9 strains were excluded in antimicrobial susceptibility test because they were lost during the experiment or were not grown in broth culture. The percentage of bacteria susceptible for each antibiotic in this study was clindamycin, 87.0% (80 of 92); tetracycline, 75.0% (69 of 92); cefuroxime axetil, 75.0% (69 of 92); amoxicillin + clavulanic acid (5:1), 71.7% (66 of 92); penicillin G, 66.3% (61 of 92); erythromycin, 66.3% (61 of 92); amoxicillin, 44.6% (41 of 92); and ciprofloxacin, 31.5% (29 of 92). The susceptibility pattern of 8 antibiotics was dependent on the host of the bacteria strains rather than the kinds of bacterial species. These results indicate that antibiotic susceptibility test should be performed when antibiotics are needed for the treatment of infected root canals.

The Ability of Anti-tumor Necrosis Factor Alpha(TNF-${\alpha}$) Antibodies Produced in Sheep Colostrums

  • Yun, Sung-Seob
    • 한국유가공학회:학술대회논문집
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    • 2007.09a
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    • pp.49-58
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    • 2007
  • Inflammatory process leads to the well-known mucosal damage and therefore a further disturbance of the epithelial barrier function, resulting abnormal intestinal wall function, even further accelerating the inflammatory process[1]. Despite of the records, etiology and pathogenesis of IBD remain rather unclear. There are many studies over the past couple of years have led to great advanced in understanding the inflammatory bowel disease(IBD) and their underlying pathophysiologic mechanisms. From the current understanding, it is likely that chronic inflammation in IBD is due to aggressive cellular immune responses including increased serum concentrations of different cytokines. Therefore, targeted molecules can be specifically eliminated in their expression directly on the transcriptional level. Interesting therapeutic trials are expected against adhesion molecules and pro-inflammatory cytokines such as TNF-${\alpha}$. The future development of immune therapies in IBD therefore holds great promises for better treatment modalities of IBD but will also open important new insights into a further understanding of inflammation pathophysiology. Treatment of cytokine inhibitors such as Immunex(Enbrel) and J&J/Centocor(Remicade) which are mouse-derived monoclonal antibodies have been shown in several studies to modulate the symptoms of patients, however, theses TNF inhibitors also have an adverse effect immune-related problems and also are costly and must be administered by injection. Because of the eventual development of unwanted side effects, these two products are used in only a select patient population. The present study was performed to elucidate the ability of TNF-${\alpha}$ antibodies produced in sheep colostrums to neutralize TNF-${\alpha}$ action in a cell-based bioassay and in a small animal model of intestinal inflammation. In vitro study, inhibitory effect of anti-TNF-${\alpha}$ antibody from the sheep was determined by cell bioassay. The antibody from the sheep at 1 in 10,000 dilution was able to completely inhibit TNF-${\alpha}$ activity in the cell bioassay. The antibodies from the same sheep, but different milkings, exhibited some variability in inhibition of TNF-${\alpha}$ activity, but were all greater than the control sample. In vivo study, the degree of inflammation was severe to experiment, despite of the initial pilot trial, main trial 1 was unable to figure out of any effect of antibody to reduce the impact of PAF and LPS. Main rat trial 2 resulted no significant symptoms like characteristic acute diarrhea and weight loss of colitis. This study suggested that colostrums from sheep immunized against TNF-${\alpha}$ significantly inhibited TNF-${\alpha}$ bioactivity in the cell based assay. And the higher than anticipated variability in the two animal models precluded assessment of the ability of antibody to prevent TNF-${\alpha}$ induced intestinal damage in the intact animal. Further study will require to find out an alternative animal model, which is more acceptable to test anti-TNF-${\alpha}$ IgA therapy for reducing the impact of inflammation on gut dysfunction. And subsequent pre-clinical and clinical testing also need generation of more antibody as current supplies are low.

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