• Title/Summary/Keyword: Internal strain

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Saprolegnia parasitica Isolated from Rainbow Trout in Korea: Characterization, Anti-Saprolegnia Activity and Host Pathogen Interaction in Zebrafish Disease Model

  • Shin, Sangyeop;Kulatunga, D.C.M.;Dananjaya, S.H.S.;Nikapitiya, Chamilani;Lee, Jehee;De Zoysa, Mahanama
    • Mycobiology
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    • v.45 no.4
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    • pp.297-311
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    • 2017
  • Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

Role of C-terminal 7 Amino Acids of N4SSB Protein in Its in vivo Activity (N4SSB 단백질의 C-말단기의 7개의 아미노산이 N4SSB 단백질의 in vivo 활성에 미치는 영향)

  • Choi, Mieyoung
    • Korean Journal of Microbiology
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    • v.34 no.4
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    • pp.248-253
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    • 1998
  • Bacteriophage N4, a lytic phage specific for Esherichia coli K12 strain encodes single-stranded DNA-binding protein, N4SSB (bacteriophage N4-coded single-stranded DNA-binding protein). N4SSB protein is originally identified as a protein required for N4 DNA replication. N4SSB protein is also required for N4 late transcription, which is catalyzed by E. coli ${\sigma}^{70}$ RNA polymerase. N4 late transcription does not occur until N4SSB protein is synthesized. Recently it is reported that N4SSB protein is essential for N4 DNA recombination. Therefore N 4SSB protein is a multifunctional protein required for N4 DNA replication, late transcription, and N4 DNA recombination. In this study, a variety of mutant N4SSB proteins containing internal deletions or substitutions were constructed to define and characterize domains important for N4 DNA replication, late transcription, and N4 DNA recombination. Test for the ill vivo activity of these mutant N4SSBs for N4 DNA replication, late transcription, and N4 DNA recombination was examined. The results suggest that C-terminal 7 amino acid residues are important for the activity of N4SSB. Three lysine residues, which are contained in this region play important roles on N4SSB activity.

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Mechanism of Metronidazole Resistance Regulated by the fdxA Gene in Helicobacter pylori. (헬리코박터 파일로리에서 fdxA 유전자에 의한 메트로니다졸 내성 조절 기전 연구)

  • Nam, Won-Hee;Lee, Sun-Mi;Kim, Eun-Sil;Kim, Jin-Ho;Jeong, Jin-Yong
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.723-727
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    • 2007
  • Resistance to metronidazole in Helicobacter pylori results from inactivation of rdxA and frxA, the chromosomal genes for a nitroreductase that normally converts metronidazole from prodrug to bactericidal agent. Two types of metronidazole susceptible strains had been found distinguishable by their apparent levels of frxA expression. Most common in the populations we had studied were strains that required only rdxA inactivation to become resistant to moderate levels of metronidazole(type I strains). The second strain type required inactivation of both frxA and rdxA to become resistance to metronidazole(type II strains): this was linked to a relatively high level of frxA gene transcription in the type II strains. The fdxA gene regulated fdxA as well as rdxA gene. Thus, to study the function of fdxA as a regulatory gene we constructed a null mutant of fdxA in H. pylori genome and identified over-and under-expressed proteins by fdxA using two-dimensional(2-D) electrophoresis and MALDI-TOP-MS. There were four over-expressed proteins in fdxA mutant; nifU-like protein(HP0221), frxA(HP0642), nonheme ferritin(HP0653), and hypothetical protein(HP0902). Three under-expressed proteins were also identified in fdxA mutant, including 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (HP0089), (3R)-hydroxymyristoyl ACP dehydratase(HP1376), and thioredoxin(HP1458).

Development of New Strains of Wolfiporia cocos for Sclerotium Formation by 2-Way Cross-Breeding (이원교배에 의해 균핵 형성하는 복령 균주 개발)

  • Ka, Kang-Hyeon;Kim, Suyeon;Park, Mi-Jeong;Jeong, Yeun Sug;Ryoo, Rhim;Jang, Yeongseon;Choi, Jong-Woon;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.49 no.3
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    • pp.405-412
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    • 2021
  • Wolfiporia cocos is an important medicinal fungus that has been used in regions of Northeast Asia including Korea, Japan, and China. W. cocos is classified in Korea into two types (red bokryeong and white bokryeong) based on the internal colors (yellow orange-pale pink and white) of the sclerotium. Generally, the W. cocos type cultivated on farms produces white sclerotium. In this study, we endeavored to select strains that form sclerotium in sawdust medium using 2-way cross-breeding among two cultivated strains and three wild strains. Monospores were isolated from the fruiting bodies of cultivated and wild strains on potato dextrose agar. Thirty-nine strains of 338 hybrid strains isolated formed sclerotia with white or yellow colors upon culture for 3 months in Pinus densiflora sawdust medium. Selection for sclerotium forming strains using sawdust culture follows a very simple and easy procedure that is presented for the first time in this paper. We plan to test selected strains in the field to aid in developing new varieties for the future.

Selection of Entomopathogenic Fungus Isaria javanica FT333 for Dual Control of Thrips and Anthracnose (총채벌레 및 고추탄저병의 동시 방제를 위한 곤충병원성 곰팡이 Isaria javanica FT333 선발)

  • Lee, Moran;Jeong, Hyeju;Kim, Jaeyoon;Kim, Dayeon;Ahn, Seung Ho;Lee, SangYeob;Han, Ji Hee
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.479-490
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    • 2018
  • Red pepper is seriously damaged by thrips (Thrips palmi) and anthracnose caused by Colletotrichum acutatum throughout its development. Because of biotic constraints, producers often depend on chemicals that are expensive and have adverse effects on the environment, operator, and beneficial insects. In addition, resistance is developed because of the repeated use of chemicals. In recent decades, the use of microorganisms in crop protection has become a credible alternative because it is eco-friendly. In this study, we aimed to select isolates with insecticidal and fungicidal activities against the pathogens that cause anthracnose and thrips. We treated T. palmi adults and juveniles with 13 strains of entomopathogenic fungi (isolated from the soil by using the insect-bait method), and 6 strains showed excellent insecticidal activity (70-100%) 5 days after the treatment. The selected isolates were cultured with C. acutatum to screen for the strain with excellent anti-fungal activities, among which an isolate FT333 showed more than 95% control efficacy against C. acutatum in vitro. The isolate was identified as Isaria javanica through its morphological characteristics and phylogenetic analysis of the ITS and ${\beta}-tubulin$ nucleotide sequences. The Isaria javanica FT333 isolate could be used effectively for dual bio-control of thrips and anthracnose during red pepper cultivation.

Suitable Conditions for Mycelial Culture of Tremella fuciformis (흰목이 균사체 배양 적합 조건 설정)

  • Lee, Eun-ji;Park, Hye-Sung;Lee, Chan-Jung;Kong, Won-Sik;Koo, Chang-Duck
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.1-12
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    • 2019
  • This study was conducted to investigate the optimum culture conditions, for mass production of Tremella fuciformis in M9 basic medium. The strain KMCC04674, used in this study was identified T. fuciformis by internal transcribed spacer (ITS). To define the optimum conditions for the mass production of T. fuciformis, we investigated the effects of different culture conditions and various nutrient sources on the fungal growth. The optimum initial pH and temperature for the fungal growth were 5.0 and $25^{\circ}C$, respectively. The optimal composition of the growth medium was 4.0% mannitol, 3.0% $NH_4H_2PO_4$, 1.0% malt extract, 1.0% glutamic acid, 5mM $CaCl_2$, and 0.5% glucanic acid.

Scale Effects of Initial Model and Material on 3-Dimensional Distinct Element Simulation (3차원 개별요소해석 시의 초기 모델 및 재료 스케일 영향)

  • Jeon, Jesung;Shin, Donghoon;Ha, Iksoo
    • Journal of the Korean GEO-environmental Society
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    • v.12 no.7
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    • pp.57-65
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    • 2011
  • Numerical simulations by three-dimensional Particle Flow Code($PFC^{3D}$, Itasca) considering distinct element method (DEM) were carried out for prediction of triaxial compression test with sand material. The effect of scale conditions for numerical model and distinct material on final prediction results was analyzed by numerical models under various scale conditions, and following observations were made from the numerical experiments. It is very useful to model the initial material condition without any porosity conversion from 2-D to 3-D DEM. Numerical experiments have shown that in all cases considered, 3D distinct element modeling could provide good agreement on stress-strain behavior, volume change and strength properties with laboratory testing results. It was important thing to assess reasonable scale ratio of numerical model and distinct elements for saving calculation time and securing calculation efficiency under condition with accuracy and appropriateness as numerical laboratory. As results of DEM simulations under various scale conditions, most of results show that shear strength properties as cohesion and internal friction angle are similar in condition of $D_{mod}/D_{gmax}$ < 10. It shows that 3-D distinct element method could be used as efficient tool to assess strength properties by numerical laboratory technique.

A Study on the Buckling Stability due to Lateral Impact of Gas Pipe Installed on the Sea-bed (해저면에 설치된 가스관의 외부충격에 의한 좌굴 안전성 검토)

  • Park, Joo-Shin;Yi, Myung-Su
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.28 no.2
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    • pp.414-421
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    • 2022
  • Subsea oil and gas exploration is increasingly moving into deeper water depths, and typically, subsea pipelines operate under high pressure and temperature conditions. Owing to the difference in these components, the axial force in the pipe is accumulated. When a pipeline is operated at a high internal pressure and temperature, it will attempt to expand and contract for differential temperature changes. Typically, the line is not free to move because of the plane strain constraints in the longitudinal direction and soil friction effects. For a positive differential temperature, it will be subjected to an axial compressive load, and when this load reaches a certain critical value, the pipe may experience vertical (upheaval buckling) or lateral (snaking buckling) movements that can jeopardize the structural integrity of the pipeline. In these circumstances, the pipeline behavior should be evaluated to ensure the pipeline structural integrity during operation in those demanding loading conditions. Performing this analysis, the correct mitigation measures for thermal buckling can be considered either by accepting bar buckling but preventing the development of excessive bending moment or by preventing any occurrence of bending.

Relation Between Shrinkage and Humidity on Lightweight Concrete and Normal Concrete by Water-Cement Ratio (물-시멘트비에 따른 경량콘크리트 및 일반콘크리트의 수축과 습도와의 관계)

  • Lee, Chang Soo;Park, Jong Hyok;Jung, Bong Jo;Choi, Young Jun
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.29 no.4A
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    • pp.385-393
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    • 2009
  • This study grasped the relationship between relative humidity in concrete and concrete shrinkage followed by pre-absorbed water of porous lightweight aggregates through measurements of concrete shrinkage and humidity and comparisons with established research results. It was showed that shrinkage reduction effect of lightweight concrete is 36% at 7 days early ages and 25% at 180 days long-term ages when water-binder ratio is 0.3. It also showed that shrinkage reduction effect is 19% at 7 days and 16% at 180 days when water-binder ratio is 0.4 and 37%, 32% when water-binder ratio is 0.5. The moisture supply effect of lightweight aggregates was remarkable at early age within 7~10 days irrespective of water-binder ratio. In case of waterbinder ratio is 0.3, the relationship between shrinkage and internal humidity of concrete has been underestimated regardless of applied existing model type and in case of water-binder ratio is 0.4, 0.5, measurement values are relatively similar with existing model equations. Finally this study did regression analyses about the relation among the humidity change and the shrinkage strain as a high-degree polynomial and derived parameters that can connect moisture movement analysis with differential shrinkage analysis in case of considering relative humidity at the time by moisture movement analysis of concrete.

Effects of Cellulolytic Microbes Inoculation During Deep Stacking of Spent Mushroom Substrates on Cellulolytic Enzyme Activity and Nutrients Utilization by Sheep (버섯부산물 퇴적발효 시 섬유소 분해균 접종이 섬유소 분해성 효소 활력과 면양의 영양소 이용성에 미치는 영향)

  • Kim, Y.I.;Jun, S.H.;Yang, S.Y.;Huh, J.W;Kwak, W.S.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.667-676
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    • 2007
  • This study was conducted to determine effects of cellulolytic microbes inoculation to sawdust-based spent mushroom substrate(SMS) during deepstacking on fermentation parameters, total microbial counts and cellulolytic enzyme activity and to on SMS nutrients utilization by sheep. For sheep metabolism trials, six sheep(ram, average 54.8kg) were fed a Control diet(70% concentrates, 15% rice straw and 15% SMS with no microbial treatment on a dry basis) and a Treatment diet(the same diet including SMS with a microbial treatment) for 2 trials. Spent mushroom substrates with or without a microbial(4 strains including 1 strain of Enterobacter ludwigii, 1 strain of Bacillus cereus and 2 strains of Bacillus subtillis) treatment (1% of SMS on wet basis) were deepstacked for 7 days. The internal temperatures in 1.2 M/T of SMS deepstacks reached to 50±5℃ within 7 days of storage. Total microbial counts remarkably decreased (P<0.05) with a deepstacking process and were not affected(P>0.05) by the microbial treatment. For fibrolytic enzyme activity, CMCase and xylanase activities were decreased(P<0.05) by a deepstacking process. After deepstacking, the microbial treatment showed about 2.5-times higher(P<0.05) for CMCase activity and about 4-times higher(P<0.05) for xylanase activity than those of the Control. Activities of ligninolytic enzymes such as laccase and MnP were not affected by the microbial treatment. The sheep fed the microbially treated SMS diet had a tendency of greater total tract digestibilities of ash(P=0.051), NFE (P=0.071), hemicellulose(P=0.087) and NDF(P=0.096) than those fed the untreated SMS diet. Nitrogen balance of sheep was not affected(P>0.05) by feeding of microbially treated SMS. Accordingly, these results indicate that cellulolytic microbes inoculation during deepstacking of SMS may improve the bio- utilization of SMS by sheep.