• Title/Summary/Keyword: secondary isolates

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Gene Mutations of 23S rRNA Associated with Clarithromycin Resistance in Helicobacter pylori Strains Isolated from Korean Patients

  • Kim, Jung-Mogg;Kim, Joo-Sung;Kim, Na-Young;Kim, Yeoung-Jeon;Kim, In-Young;Chee, Young-Joon;Lee, Chul-Hoon;Jung, Hyun-Chae
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1584-1589
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    • 2008
  • Although resistance of Helicobacter pylori to clarithromycin is a major cause of failure of eradication therapies, little information is available regarding gene mutations of clarithromycin-resistant primary and secondary H. pylori isolates in Korea. In the present study, we examined gene mutations of H. pylori 238 rRNA responsible for resistance to clarithromycin. DNA sequences of the 238 rRNA gene in 21 primary clarithromycin-resistant and 64 secondary clarithromycin-resistant strains were determined by PCR amplification and nucleotide sequence analyses. Two mutations of the 238 rRNA gene, A2143G and T2182C, were observed in primary clarithromycin-resistant isolates. In secondary isolates, dual mutation of A2143G+T2182C was frequently observed. In addition, A2143G+T2182C+ T2190C, A2143G+T2182C+C2195T, and A2143G+T2182C+A2223G were observed in secondary isolates. Furthermore, macrolide binding was tested on purified ribosomes isolated from T2182C or A2143C mutant strains with $[^{14}C]$erythromycin. Erythromycin binding increased in a dose-dependent manner for the susceptible strain but not for the mutant strains. These results indicate that secondary isolates show a greater variety of 238 rRNA gene mutation types than primary isolates, and triple mutations of secondary isolates are associated with A2143G+T2182C in H. pylori isolated from Korean patients.

Production and Identification of Secondary Metabolite Gliotoxin-Like Substance Using Clinical Isolates of Candida spp.

  • Noorulhuda Ojaimi Mahdi, Al-Dahlaki;Safaa Al-Deen Ahmed Shanter, Al-Qaysi
    • Microbiology and Biotechnology Letters
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    • v.50 no.4
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    • pp.488-500
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    • 2022
  • Most fungal infections by opportunistic yeast pathogens such as Candida spp. are the major causes of morbidity and mortality in patients with lowered immune. Previous studies have reported that some strains of Candida secret secondary metabolites play an important role in the decreasing of immunity in the infected patient. In this study, 110 Candida spp. were isolated from different clinical specimens from Baghdad hospitals. Candida isolates were identified by conventional methods, they were processed for Candida speciation on CHROMagar. The results of identification were confirmed by internal transcribed spacer (ITS) sequencing. Phylogenetic trees were analyzed with reference strains deposited in GenBank. Antifungal susceptibility testing was evaluated by the disc diffusion method and performed as recommended by the Clinical and Laboratory Standard Institute (CLSI) M44-A document. Candida isolates investigated produce secondary metabolites gliotoxin with HPLC technique and quantification. Out of 110 Candida isolates, C. albicans (66.36%) was the most frequent isolate, followed by the isolates of C. tropicalis (10.9%) and C. glabrata (6.36%) respectively. Concerning the antifungal susceptibility test, Candida isolates showed a high level of susceptibility to Miconazole (70.9%), Itraconazole (68.2%), and Nystatine (64.5%). The ability of obtained isolates of Candida spp. to produce gliotoxin on RPMI medium was investigated, only 28 isolates had the ability to secret this toxin in culture filtrates. The highest concentrations were detected in C. albicans (1.048 ㎍/ml). Gliotoxin productivity of other Candida species was significantly lower. The retention time for gliotoxin was approximately 5.08 min.

Nucleotide Sequence Analysis and Secondary Structure Modeling of the 3'-Noncoding Regions of Two Korean Strains of Turnip Mosaic Virus (순무 모자이크 바이러스 두 한국계통의 3' 말단 비번역부위에 대한 염기서열분석 및 2차구조 모델링)

  • 최장경;류기현;최국선;박원목
    • Korean Journal Plant Pathology
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    • v.11 no.3
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    • pp.271-277
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    • 1995
  • The RNA nucleotide sequences of the 3/-noncoding regions (3'-NCRs) of two Korean strains of turnip mosaic virus (TuMV), Ca and cqs, have been determined from their cDNA clones that encompassed the 3'-terminal regions of the viral genomic RNAs. The 3'-NCRs of both strains were 209 nucleotides long, terminated with GAC residues and poly (A) tails. The potential polyadenylational signal motif, UAUGU, was located 140 nucleotides upstream from the poly (A) tail in each of the virus. A highly conserved hexanucleotide sequence [A G U G A/U G/C], which was common in the 3'-NCRs of the potyvirus RNAs, was also found at the regions of 119 bases upstream from the 3'-end. Comparison of the 3'-NCRs of the two Korean isolates with those of four strains from Canada, China and Japan showed significantly identical genotypes (94.3∼99.5%). The secondary structure of three loops with long stems was found within the 3'-NCRs by sequence analysis. The substituted bases in the region among the six TuMV strains did not alter their secondary structures. Length of the 3'-NCRs of the know 11 potyviral RNAs and TuMV RNAs was different from one another and their nucleotide sequences showed 55.7% to 24.0% of homology. The 3'-NCR, therefore, is considered to be useful for phylogenetic studies in potyviruses.

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Isolation and Characterization of Bacteriolytic Wild Myxobacteria (용균성 야생 점액세균의 분리)

  • 박수연;이봉수;김지훈;이차율;장은혜;조경연
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.218-223
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    • 2004
  • Myxobacteria are Gram-negative soil bacteria known to be a rich source of potentially useful secondary metabolites. We have isolated 204 strains of bacteriolytic myxobacteria from soil samples collected in Korea and determined their 16S rRNA sequences. Sequence analysis of the partially determined 16S rRNA sequences has suggested that 132 isolates (65% of total isolates) belong to the genus Myxococcus and 59 isolates (29% of total isolates) belong to the genus Corallococcus. Meanwhile, 4 isolates appear to be Archangium spp. and the other 4 isolates appear to be Stigmatella spp. Genera of the remained 5 isolates have not been identified because their 16S rRNA sequences are distantly related to those of known myxobacteria.

Effects of gamma irradiation on the functional and structural characteristics of soy protein isolates (방사선 조사가 분리대두단백의 기능적, 구조적 특성에 미치는 영향)

  • 오경남;이숙영
    • Korean journal of food and cookery science
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    • v.20 no.3
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    • pp.256-264
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    • 2004
  • The effects of irradiation on the functional and structural characteristics of soy protein isolates were studied. Soymilk was irradiated at 1, 5, and l0kGy, after which soy protein isolates were prepared. The functional properties of soy protein isolates were examined including solubility, emulsion capacity and stability, foam capacity and stability, structural properties as represented by SDS-PAGE pattern, and secondary and tertiary structures. The solubility and emulsion capacity were increased by radiation treatment at 1kGy however the values were adversely affected again as dosage was increased above 5kGy. As irradiation dosage increased, an increase of foaming capacity at 1kGy and a decreasing turnover afterwards were also noted in foaming capacity, although the differences were not statistically significant. The SDS-PAGE pattern showed fragmentation and aggregation of protein molecules as affected by irradiation in proportion to the dosage increase. The results of CD and fluorescence spectroscopy revealed increased aperiodic structure contents with the dosage increase. It was assumed that irradiation dosagefrom 5 to l0kGy could initiate minimal denaturation of protein in various foods compared to general heat treatment.

A highly efficient computational discrimination among Streptococcal species of periodontitis patients using 16S rRNA amplicons

  • Al-Dabbagh, Nebras N.;Hashim, Hayder O.;Al-Shuhaib, Mohammed Baqur S.
    • Korean Journal of Microbiology
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    • v.55 no.1
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    • pp.1-8
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    • 2019
  • Due to the major role played by several species of Streptococcus in the etiology of periodontitis, it is important to assess the pattern of Streptococcus pathogenic pathways within the infected subgingival pockets using a bacterial specific 16S rRNA fragment. From the total of 50 patients with periodontitis included in the study, only 23 Streptococcal isolates were considered for further analyses, in which their 16S rRNA fragments were amplified and sequenced. Then, a comprehensive phylogenetic tree was constructed and in silico prediction was performed for the observed Streptococcal species. The phylogenetic analysis of the subgingival Streptococcal species revealed a high discrimination power of the 16S rRNA fragment to accurately identify three groups of Streptococcus on the species level, including S. salivarius (14 isolates), S. anginosus (5 isolates), and S. gordonii (4 isolates). The employment of state-of-art in silico tools indicated that each Streptococcal species group was characterized with particular transcription factors that bound exclusively with a different 16S rRNA-based secondary structure. In conclusion, the observed data of the present study provided in-depth insights into the mechanism of each Streptococcal species in its pathogenesis, which differ in each observed group, according to the differences in the 16S rRNA secondary structure it takes, and the consequent binding with its corresponding transcription factors. This study paves the way for further interventions of the in silico prediction, with the main conventional in vitro microbiota identification to present an interesting insight in terms of the gene expression pattern and the signaling pathway that each pathogenic species follows in the infected subgingival site.

Molecular Characterization of Pseudomonas aeruginosa Isolates Resistant to All Antimicrobial Agents, but Susceptible to Colistin, in Daegu, Korea

  • Lee, Yoo-Chul;Ahn, Byung-Jun;Jin, Jong-Sook;Kim, Jung-Uk;Lee, Sang-Hwa;Song, Do-Young;Lee, Won-Kil;Lee, Je-Chul
    • Journal of Microbiology
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    • v.45 no.4
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    • pp.358-363
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    • 2007
  • Multi-drug resistant Pseudomonas aeruginosa has been implicated in a variety of serious therapeutic problems in clinical environments. Among the 968 P. aeruginosa isolates obtained from two hospitals in Daegu, Korea, we acquired 17 isolates that were resistant to all available tested antimicrobial agents, with the exception of colistin (colistin-only sensitive). We characterized the antimicrobial susceptibilities, $metallo-{\beta}-lactamases$, and epidemiological relatedness among the colistin-only sensitive P. aeruginosa isolates. All colistin-only sensitive isolates were positive in the modified Hodge test and imipenem-EDTA synergy test, thereby indicating the production of $metallo-{\beta}-lactamases$. 11 isolates from the secondary hospital and six isolates from the tertiary teaching hospital harbored $bla_{VIM-2}$ and $bla_{IMP-1}$, respectively. The pulsed-field gel electrophoretic analysis of the SpeI-digested DNA from P. aeruginosa isolates indicated that two different clones of colistin-only sensitive P. aeruginosa originated from each hospital, and had spread within the hospital environment. Overall, colistin-only sensitive P. aeruginosa was detected in Korea for the first time, but no pan-drug resistant bacteria were identified. Nationwide surveillance is required in order to monitor the emergence of colistin-only sensitive or pan-drug resistant bacteria.

Cytochalasin E Production by Rosellinia necatrix and Its Pathogenicity on Apple (사과흰날개무늬병균의 Cytochalasin E 독소 생산과 병원성)

  • Lee, Dong-Hyuk;Choi, Kyung-Hee;Uhm, Jae-Youl
    • Research in Plant Disease
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    • v.15 no.1
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    • pp.46-50
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    • 2009
  • Cytochalasin E (CE) is a secondary metabolite secreted by Rosellinia necatrix, caused by white root rot, and has toxicity to apple as a toxin during disease progress. This study was conducted to demonstrate the relationship between the production of CE and its pathogenicity. CE producing isolates and non-producing isolates of R. nectatrix were isolated from the mycerial mat of diseased roots and was detected on that using a TLC and HPLC analysis and in vivo pathogenicity test. CE non-producing isolates were not pathogenic to apple roots and not detected CE by TLC and HPLC analysis. It was shown that the production of CE was related to the pathogenicity of R. nectatrix.

Profiles of Bacillus spp. Isolated from the Rhizosphere of Suaeda glauca and Their Potential to Promote Plant Growth and Suppress Fungal Phytopathogens

  • Lu, Ping;Jiang, Ke;Hao, Ya-Qiao;Chu, Wan-Ying;Xu, Yu-Dong;Yang, Jia-Yao;Chen, Jia-Le;Zeng, Guo-Hong;Gu, Zhou-Hang;Zhao, Hong-Xin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1231-1240
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    • 2021
  • Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.

Screening of Endophytic Fungal Isolates Against Raffaelea quercus-mongolicae Causing Oak Wilt Disease in Korea

  • Nguyen, Manh Ha;Yong, Joo Hyun;Sung, Han Jung;Lee, Jong Kyu
    • Mycobiology
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    • v.48 no.6
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    • pp.484-494
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    • 2020
  • Oak wilt disease caused by Raffaelea quercus-mongolicae has emerged obviously in Korea. We selected antifungal isolates against R. quercus-mongolicae among 368 endophytic fungal isolates from different parts of oak and pine trees. The experiment was conducted in the primary and secondary screenings by dual culture test. The antifungal activity of the selected isolates was assessed in culture filtrate test based on the inhibition rates in mycelial growth, sporulation, and spore germination of oak wilt fungus. Five isolates, E089, E199, E282, E409 and E415, showed strong antifungal activity in culture filtrate test, and their antifungal activity decreased on the culture media supplemented with heated culture filtrate. Higher mycelial growth inhibitions on the unheated media were recorded in E409 (Colletotrichum acutatum), E089 (Daldinia childiae), E415 (Alternaria alternata) and E199 (Daldinia childiae) with the inhibition rates of 79.0%, 70.1%, 68.9% and 64.5%, respectively. These isolates also had the higher sporulation inhibitions on unheated media with the rates of 96.8%, 84.2%, 82.8% and 80.5%, respectively. The spore germination of the oak wilt fungus was completely inhibited by E282 (Nectria balsamea) on both unheated and heated media. These results showed that a higher number of potent antifungal isolates against oak wilt fungus was isolated from the petiole compared to the other parts. This study could contribute to the development of biological control approaches for the management of oak wilt disease caused by R. quercus-mongolicae.