• Title/Summary/Keyword: acid-resistance

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The Attenuation Mechanism and Live Vaccine Potential of a Low-Virulence Edwardsiella ictaluri Strain Obtained by Rifampicin Passaging Culture

  • Shuyi Wang;Jingwen Hao;Jicheng Yang;Qianqian Zhang;Aihua Li
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.167-179
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    • 2023
  • The rifampicin-resistant strain E9-302 of Edwardsiella ictaluri strain 669 (WT) was generated by continuous passage on BHI agar plates containing increasing concentrations of rifampicin. E9-302 was attenuated significantly by 119 times to zebrafish Danio rerio compared to WT in terms of the 50% lethal dose (LD50). Zebrafish vaccinated with E9-302 via intraperitoneal (IP) injection at a dose of 1 × 103 CFU/fish had relative percentage survival (RPS) rates of 85.7% when challenged with wild-type E. ictaluri via IP 14 days post-vaccination (dpv). After 14 days of primary vaccination with E9-302 via immersion (IM) at a dose of 4 × 107 CFU/ml, a booster IM vaccination with E9-302 at a dose of 2 × 107 CFU/ml exhibited 65.2% RPS against challenge with wild-type E. ictaluri via IP 7 days later. These results indicated that the rifampicin-resistant attenuated strain E9-302 had potential as a live vaccine against E. ictaluri infection. A previously unreported amino acid site change at position 142 of the RNA polymerase (RNAP) β subunit encoded by the gene rpoB associated with rifampicin resistance was identified. Analysis of the whole-genome sequencing results revealed multiple missense mutations in the virulence-related genes esrB and sspH2 in E9-302 compared with WT, and a 189 bp mismatch in one gene, whose coding product was highly homologous to glycosyltransferase family 39 protein. This study preliminarily explored the molecular mechanism underlying the virulence attenuation of rifampicin-resistant strain E9-302 and provided a new target for the subsequent study of the pathogenic mechanism of E. ictaluri.

Performance Evaluation of Concrete Bench Flume Using Industrial by Products (산업부산물을 이용한 콘크리트 벤치플룸의 성능평가)

  • Jae-Ho Jung
    • Journal of the Korean Recycled Construction Resources Institute
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    • v.11 no.3
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    • pp.276-281
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    • 2023
  • Water pipes manufactured using existing Portland cement suffer from the problem of rapid deterioration and reduced durability due to the hydration product of cement being vulnerable to acids. Therefore, in this study, water pipes were manufactured using slag and fly ash, which are industrial by-products from various industries, and their characteristics were analyzed. As a result of the experiment, slump in unhardened concrete tended to increase due to the ball bearing action of fly ash, and the amount of air was reduced due to unburned coal, indicating that measures for frost resistance were needed. In addition, the initial strength of the compressive strength was increased through steam curing, and the results were equal to or better than OPC when mixing more than 50 % of slag. The acid resistance results showed that the mass reduction rate was less than 5 %, showing excellent durability performance, and the bending failure load of the water pipe also exceeded the KS standards, so it is judged to be commercializable.

In Vitro Screening of Tannic Acid-based Eco-friendly Farming Material (notice no. 2-4-064) against Plant Pathogenic Bacteria (탄닌산을 함유한 친환경농자재(공시번호 2-4-064)의 식물병원세균 기내 억제효과)

  • Han, Kyu Suk;Ju, Ho-Jong;Hong, Jin Sung;Chung, Jong-Sang;Park, Duck Hwan
    • Korean Journal of Organic Agriculture
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    • v.24 no.4
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    • pp.945-955
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    • 2016
  • To date, chemical managements of plant bacterial diseases are complicated by limitations on use of antibiotics in agriculture, antibiotic resistance development, and limited efficacy of alternative control agents. In this study, thus, we performed screening of eco-friendly farming material (notice no. 2-4-064) containing tannic acid as new antibacterial-activity against 7 plant bacterial pathogens: Pectobacterium carotovorum subsp. carotovorum (Pcc), Ralstonia solanacearum (Rs), Acidovorax avenae subsp. citrulli (Aac), Xanthomonas cirti pv. citri (Xcc), Erwinia pyrifoliae (Ep), Clavibacter michiganensis subsp. michiganensis (Cmm), and Streptomyces scabies (Sc), Initial screening of antibacterial effects of eco-friendly farming material was performed using the paper disk diffusion method and co-cultivation in broth culture. Tannic acid based eco-friendly farming material showed inhibitory effect against Pcc, Rs, Aac, Xcc, Cmm, and Ss, whereas it did not show inhibition zone against Ep. However, when it tested by co-cultivation in broth culture, it showed strong inhibition effect against all pathogens that declined growth curve compared to bacterial pathogen only. Interestingly, when we observed morphological changes on those pathogens by SEM, cell morphologies of 7 pathogens were slightly changed that seem to be malfunction in their cell envelope.

Effects of Hypovolemic Hypotension on Cardiopulmonary Functions and Acid-Base Balance (출혈성 쇼크가 심폐기능 및 산.염기평형에 미치는 영향)

  • Sou, Won-Young;Lee, Sung-Haing
    • Journal of Chest Surgery
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    • v.6 no.2
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    • pp.131-142
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    • 1973
  • Studies of cardiopulmonary function and acid-base balance were performed on 29 dogs during control period, during oligemic hypotension and following return of blood to the animals. Intravenous morphine and local anesthesia were used. Fifteen of the 29 animals survived the complete experiment. The 14 animals that failed to survive the experimental period died between 15 to 90 minutes after the onset of bleeding. The results were as follows. 1. The heart rate increased after the onset of bleeding and failed to return to control level following reinfusion. Stroke volume decreased markedly after bleeding and failed to recover after return of blood from the reservoir. Cardiac output also decreased during oligemic hypotension and was maintained at this level after re-infusion. Total peripheral resistance decreased significantly immediately after bleeding, however it increased soon over the pre-bleeding level. Central venous pressure decreased after the onset of bleeding and remained at lower level for the rest of the experimental period. Arterial blood pressure, clown to 40-45 mmHg by acute hemorrhage, was elevated near to control level. Left ventricular work decreased tremendously during oligemic hypotension and failed to return to control level with the re-infusion of blood. Hematocrit value showed no significant decrease after bleeding and increased after re-infusion. Hemoglobin decreased after the onset of bleeding and recovered to control value after re-infusion. 2. The respiratory rate fell rapidly after bleeding from 124 to 29 and remained at this lower level for the remainder of the experiment. The tidal volume increased after bleeding and was maintained at this level for the remainder of the experiment. The respiratory minute volume showed no significant changes throughout the experimental period. Oxygen consumption fell lightly in all animals during oligemic hypotension and returned to normal levels following re-infusion. Arterial oxygen content and arterial oxygen saturation decreased following bleeding and the values returned to normal levels after the return of blood from the reservoir The arterio-venous oxygen difference increased after the onset of bleeding. It failed to return to normal values following re-infusion. Arterial $Pco_2$ decreased in all animals after the beginning of the bleeding. Partial pressure of $Co_2$ continued to fall until re-infusion, after which the values returned toward normal. Animals became acidotic. The pH fell to lower level following bleeding. Lactic acid and lactate: pyruvate ratio also increased during same period. Arterial pH and lactic acid failed to return to control value and lactate: pyruvate ratio increased more after re-infusion. Sodium bicarbonate decreased after bleeding and returned to control value following re-infusion.

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Establishment of Baseline Sensitivity of Phytophthora capsici Causing Pepper Phytophthora Blight to Carboxylic Acid Amide Fungicides (Carboxylic acid amide계 살균제에 대한 고추 역병균 Phytophthora capsici의 감수성 기준 설정)

  • Kim, Jin-Ho;Kim, Joo-Hyung;Lee, Kyeong-Hee;Rho, Chang-Woo;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.456-462
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    • 2010
  • Baseline sensitivity to benthiavalicarb, iprovalicarb and dimethomorph included into carboxylic acid amide (CAA) group was evaluated in 180 isolates of Phytophthora capsici over 4 years from 2005 to 2008. $EC_{50}$ (effective concentration inhibiting mycelial growth by 50%) value of benthiavalicarb ranged from $0.015{\mu}g\;mL^{-1}$ to $0.049{\mu}g\;mL^{-1}$ with a mean of $0.033{\mu}g\;mL^{-1}$. The mean values of $EC_{50}$ of iprovalicarb and dimethomorph were 0.411 (0.197 - 0.556) ${\mu}g\;mL^{-1}$ and 0.271 (0.101 - 0.798) ${\mu}g\;mL^{-1}$, respectively. Although there was no increasing tendency in $EC_{50}$of benthiavalicarb and iprovalicarb during 4 years, $EC_{50}$ of dimethomorph was increased gradually by laps of time. There was no cross-resistance between each fungicide used in this study and metalaxyl. Among fungicides included into CAA group, there was a positive correlation between benthiavalicarb and iprovalicarb, and between dimethomorp and mandipropamid.

Studies on the Production of Acid Resistant Amylase by Molds (part 1) -Isolation and its Cultural pH- (Asp, 속균(屬菌)에 의(依)한 내산성(耐酸性) Amylase 생산(生産)에 관(關)한 연구(硏究) (제 I 보) (第 I 報) -분리(分離) 및 배양(培養) pH에 대(對)하여-)

  • Park, Yoon-Choong;Lee, Suk-Kun
    • Applied Biological Chemistry
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    • v.9
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    • pp.91-96
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    • 1968
  • 1. Two strains of Aspergilli producing acid resistant amylase were isolated from air. The strain $U^{-1}$ and $U^{-2}$ were similar to Asp. usamii tan type mutant in the morphological characteristics. 2. The influence of cultural pH for production of amylase were investigated in the suface culture, it was different each other in the cultural optimum initial pH. The activity of acid resistant amylase produced by strain $U^{-1}$ was higher than that of strain $U^{-2}$. The acid resistance was increased by culturing in lower pH. 3. The lower initial pH ($U^{-1}$ pH 4.0, $U^{-2}$ pH 2.0) was better than neutral (pH 6.0) for the production of amylase and growth of them. The effect of low initial pH for the production of amylase was not to accelerate but caused by change of pH in the cultural process, in the range of this experiment.

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Characterization of Noble AmpC-Type $\beta$-Lactamases Among Clinical Isolates Using New Expression/Secretion Vector (발현ㆍ분비 벡터 및 임상 균주가 생성하는 신규 AmpC-type $\beta$-lactamase의 특성)

  • 정하일;성광훈;이정훈;장선주;이상희
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.104-110
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    • 2004
  • To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.

Comparative Evaluation of Probiotic Activities of Bifidobacterium longum MK-G7 with Commercial Bifidobacteria Strains

  • Jung, Hoo-Kil;Kim, Eung-Ryool;Ji, Geun-Eog;Park, Jong-Hyun;Cha, Seong-Kwan;Juhn, Suk-Lak
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.147-153
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    • 2000
  • This study was conducted to compare probiotic activities and physiological functions of Bifidobacterium longum Mk-G7 with weveral commercial and type strains of bifidobacteria. bif. longum MK-G7 showed the highest acid tolerance against HCl and acetic acid, whereas bif. infantis Y-1 showed the lowest acid tolerance and more than 4 log cycles of viable cell count decreased due to acid injuty. Viable cell counts of bifidobacteria strains decreased more than 1.5 log cycles owing to oxygen toxicity, with the exception of Bif. longum MK-G7, Bif. infantis Y-2, Bif. longum Y-3, Bif. longum Y-6, and Bif. longum RD-13 showed the highest bile tolerance, whereas Bif. longum MK-G7 showed a medium level of bile tolerance. Only Bif. longum MK-G7 howed much higher antibiotic resistance against both tetracycline and penicillin-G in the MIC(minimum inhibitory concentration) level of 24.8 mg/I and 0.52mg/I, respectively. Bif longum Y-6, and Bif. bifidum ATCC 29539 showed more than 80% of anti-mutagenicity against NQO(4-nitroquinolinel-oxide). Since the production of cytokines such as $TNF(tumor necrosis factor)-{\alpha}$ and IL (interleukin)-6, and NO(nitric oxide) in the macrophage cell line Raw 264.7 cells increased as Bif. longum MK-G7 cell concentration increased, ti was suggested that Bif. longum MK-G7 is able to enhance immunopotentiating activity in vitro. When freeze-dred Bif. longum MK-G7 was administered to mice at the dose of 1,2,4, and 6 g/kg of body weight, all of the mice survived in all feeding groups, proving the GRAS(generally recognized as safe) status of Bif. longum MK-G7. When fermented milk containing Bif. longum MK-G7 was administered to human volunteers, viable cell count of total bifidobacteria and anaerobes in the feces increased up to 0.5 log cycles more than before the administration. In particular, Bif. logum MK-G7 ingibited the growth of Bacteroides at the level of 1.0-1.5 log cycles.

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Downregulation of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in human keratinocytes by melanogenic inhibitors

  • Ahn, Kwang-Seok;Lee, Jinseon;Kim, Yeong-Shik
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.780-803
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    • 2003
  • Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.

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Leaching Kinetics of Praseodymium in Sulfuric Acid of Rare Earth Elements (REE) Slag Concentrated by Pyrometallurgy from Magnetite Ore

  • Kim, Chul-Joo;Yoon, Ho-Sung;Chung, Kyung Woo;Lee, Jin-Young;Kim, Sung-Don;Shin, Shun Myung;Kim, Hyung-Seop;Cho, Jong-Tae;Kim, Ji-Hye;Lee, Eun-Ji;Lee, Se-Il;Yoo, Seung-Joon
    • Korean Chemical Engineering Research
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    • v.53 no.1
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    • pp.46-52
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    • 2015
  • A leaching kinetics was conducted for the purpose of recovery of praseodymium in sulfuric acid ($H_2SO_4$) from REE slag concentrated by the smelting reduction process in an arc furnace as a reactant. The concentration of $H_2SO_4$ was fixed at an excess ratio under the condition of slurry density of 1.500 g slag/L, 0.3 mol $H_2SO_4$, and the effect of temperatures was investigated under the condition of 30 to $80^{\circ}C$. As a result, praseodymium oxide ($Pr_6O_{11}$) existing in the slag was completely converted into praseodymium sulfate ($Pr_2(SO_4)_3{\cdot}8H_2O$) after the leaching of 5 h. On the basis of the shrinking core model with a shape of sphere, the first leaching reaction was determined by chemical reaction mechanism. Generally, the solubility of pure REEs decreases with the increase of leaching temperatures in sulfuric acid, but REE slag was oppositely increased with increasing temperatures. It occurs because the ash layer included in the slag is affected as a resistance against the leaching. By using the Arrhenius expression, the apparent activation energy of the first chemical reaction was determined to be $9.195kJmol^{-1}$. In the second stage, the leaching rate is determined by the ash layer diffusion mechanism. The apparent activation energy of the second ash layer diffusion was determined to be $19.106kJmol^{-1}$. These relative low activation energy values were obtained by the existence of unreacted ash layer in the REE slag.