• The Plant Pathology Journal
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• 제24권3호
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• pp.337-351
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• 2008

Host-pathogen interaction in rice bacterial blight pathosystem was analyzed for a better understanding of their relationship and recognition of stable pathogenicity among the populations of Xanthomonas oryzae pv. oryzae. A total number of 52 bacterial strains isolated from diseased leaf samples collected from 12 rice growing states and one Union Territory of India, were inoculated on 16 rice varieties, each possessing known genes for resistance. Analysis of variance revealed that the host genotypes(G) accounted for largest(78.4%) proportion of the total sum of squares(SS), followed by 16.5% due to the pathogen isolates(I) and 5.1% due to the $I{\times}G$ interactions. Application of the Additive Main effects and Multiplicative Interaction(AMMI) model revealed that the first two interaction principal component axes(IPCA) accounted for 66.8% and 21.5% of the interaction SS, respectively. The biplot generated using the isolate and genotypic scores of the first two IPCAs revealed groups of host genotypes and pathogen isolates falling into four sectors. A group of five isolates with high virulence, high absolute IPCA-1 scores, moderate IPCA-2 scores, low AMMI stability index '$D_i$' values and minimal deviations from additive main effects displayed in AMMI biplot as well as response plot, were identified as possessing stable pathogenicity across 16 host genotypes. The largest group of 27 isolates with low virulence, small IPCA-1 as well as IPCA-2 scores, low $D_i$ values and minimal deviations from additive main effect predictions, possessed stable pathogenicity for low virulence. The AMMI analysis and biplot display facilitated in a better understanding of the host-pathogen interaction, adaptability of pathogen isolates to specific host genotypes, identification of isolates showing stable pathogenicity and most discriminating host genotypes, which could be useful in location specific breeding programs aiming at deployment of resistant host genotypes in bacterial blight disease control strategies.

• Plant Biotechnology Reports
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• 제5권1호
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• pp.53-60
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• 2011

Artemisinin is effective against both chloroquine-resistant and -sensitive strains of Plasmodium species. However, the low yield of artemisinin from cultivated and wild plants is a serious limitation to the commercialization of this drug. Optimization of artemisinin yield either in vivo or in vitro is therefore highly desirable. To this end, we have overexpressed the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR) gene (hmgr) from Catharanthus roseus L. in Artemisia annua L. and analyzed its influence on artemisinin content. PCR and Southern blot analyses revealed that the transgenic plants showed stable integration of the foreign hmgr gene. The reverse transcriptase-PCR results suggested that the hmgr was expressed at the transcriptional level in transgenic lines of Artemisia annua L., while the high-performance liquid chromatography analysis showed that artemisinin content was significantly increased in a number of the transgenic lines. Artemisinin content in one of the A. annua transgenic lines was 38.9% higher than that in non-transgenic plants, and HMGR enzyme activity in transgenic A. annua L. was also higher than that in the non-transgenic lines.

• 한국물환경학회지
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• 제32권6호
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• pp.628-648
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• 2016

Increased attention has been paid to the presence of veterinary antibiotics in various environmental matrices due to their toxicological behavior in the ecosystem and development of antibiotic-resistant strains of pathogenic bacteria. In the this review, 37 target antimicrobials were selected based on annual sales of antibiotics for livestock in South Korea 2014. Also, extraction and clean-up methods for the determination of the antibiotic residues in liquid samples including water, milk, and honey were comprehensively reviewed in the literature. Solid-phase extraction (SPE) was commonly used as a pre-treatment method for the samples. Most of the analytes were extracted in acidic conditions (2.5~4.0) except for aminoglycosides, which were extracted in neutral conditions (7.0~8.0). ${\beta}-Lactams$ showed the highest recoveries in neutral pH due to their degradation characteristics in acidic media. Starta-X, Oasis HLB, and Oasis MCX were frequently applied as an SPE cartridge and Oasis HLB showed the highest recoveries for the majority of antibiotic classes. The homogenized honey and milk were extracted by mixing with acids for deproteinization. Solids and other interfering substances in the extract were eliminated by centrifugation followed by membrane filtration or SPE before injection into HPLC.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8883-8886
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• 2014

Background: Tetracycline is an antibiotic widely used for the treatment of Helicobacter pylori infection, but its effectiveness is decreasing due to increasing bacterial resistance. The aim of this study was to investigate the occurrence of 16S rRNA mutations associated with resistance or reduced susceptibility to tetracycline ofHelicobacter pylori by real-time PCR (RT-PCR) assays from culture. Materials and Methods: Tetracycline susceptibility and minimal inhibition concentration (MIC) was determined by the Epsilometer test (Etest) method. A LightCycler assay developed to detect these mutations was applied to DNA extracted from culture. The 16S rRNA of these isolates was sequenced and resistance-associated mutations were identified. From 104 isolates of H. pylori examined, 11 showed resistance to tetracycline. Results: LightCycler assay was applied to DNA extracted from 11 tetracycline-susceptible and 11 tetracycline resistance H. pylori isolates. In our study the sequencing of the H. pylori wild types in 16 s rRNA gene were AGA 926-928 with MIC (0.016 to $0.5{\mu}g/ml$), while the sequencing and MIC for resistant were GGA and AGC, (0.75 to $1.5{\mu}g/ml$), respectively. Also we found a novel mutation in 2 strains with $84^{\circ}C$ as their melting temperatures and exhibition of an A939C mutation. Conclusions: We conclude that real-time PCR is an excellent method for determination of H. pylori tetracycline resistance related mutations that could be used directly on biopsy specimens.

• Clinical and Experimental Pediatrics
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• 제60권7호
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• pp.221-226
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• 2017

Purpose: Escherichia coli sequence type (ST) 131, a multidrug-resistant clone causing extraintestinal infections, has rapidly become prevalent worldwide. However, the epidemiological and clinical features of pediatric infections are poorly understood. We aimed to explore the characteristics of ST131 Escherichia coli isolated from Korean children with urinary tract infections. Methods: We examined 114 uropathogenic E. coli (UPEC) isolates from children hospitalized at Chung-Ang University Hospital between 2011 and 2014. Bacterial strains were classified into STs by partial sequencing of seven housekeeping genes (adk, fumC, gyrB, icd, mdh, purA, and recA). Clinical characteristics and antimicrobial susceptibility were compared between ST131 and non-ST131 UPEC isolates. Results: Sixteen UPEC isolates (14.0%) were extended-spectrum ${\beta}-lactamase$ (ESBL)-producers; 50.0% of ESBL-producers were ST131 isolates. Of all the isolates tested, 13.2% (15 of 114) were classified as ST131. There were no statistically significant associations between ST131 and age, sex, or clinical characteristics, including fever, white blood cell counts in urine and serum, C-reactive protein, radiologic abnormalities, and clinical outcome. However, ST131 isolates showed significantly lower rates of susceptibility to cefazolin (26.7%), cefotaxime (40.0%), cefepime (40.0%), and ciprofloxacin (53.3%) than non-ST131 isolates (65.7%, 91.9%, 92.9%, and 87.9%, respectively; P<0.001 for all). ESBL was more frequently produced in ST131 (53.3%) than in non-ST131 (8.1%) isolates (P<0.01). Conclusion: ST131 E. coli isolates were prevalent uropathogens in children at a single medical center in Korea between 2011 and 2014. Although ST131 isolates showed higher rates of antimicrobial resistance, clinical presentation and outcomes of patients were similar to those of patients infected with non-ST131 isolates.

• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.778-783
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• 1999

Campylobacter is a pathogen for both humans and animals that can be transferred from animals to humans. Four isolates, which grew under 5-10% $CO_2$ and had small and translucent colonies, were obtained from swine gastric mucosa and characterized using various methods. These bacteria were gram negative, spirally shaped with round ends. One or two non-sheathed polar flagella were observed under electron microscopy. A PCR with species-specific protein (SSP) primers for 16S rRNA gene in Campylobacter produced a typical 462 bp fragment. The isolates had various biochemical and molecular characteristics which differentiated them from other Campylobacters. The isolates were catalase and oxidase positive, urease (rapid) negative, nitrate reduction positive, indoxyl acetate hydrolysis positive, y-glutamyl transpeptidase negative, and alkaline phosphatase negative. All four isolates showed growth at $37^{\circ}C{\;}and{\;}42^{\circ}C{\;}but{\;}not{\;}at{\;}25^{\circ}C$, were resistant to cephalotin and cefoperazone, and susceptible to carbenicillin. The isolates showed various results in the reduction of chloride to triphenyl tetrazolium (TTC) and a susceptibility to nalidixic acid. Western blot analysis of these isolates with antiserum raised against one isolate showed different patterns from those of reference strains. A dendrogram drawn with the RAPD results showed that these isolates belonged to a new Campylobacter spp. group different from those of C. jejuni, C. doylei, C. lari, and C. coli.

• Journal of Microbiology
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• 제43권2호
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• pp.172-178
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• 2005

To determine the prevalence and genotypes of $\beta$-lactamases among clones of a metagenomic library from the cold-seep sediments of Edison seamount (10,000 years old), we performed pulse-field gel electrophoresis, antibiotic susceptibility testing, pI determination, and DNA sequencing analysis. Among the 8,823 clones of the library, thirty clones produced $\beta$-lactamases and had high levels of genetic diversity. Consistent with minimum inhibitory concentration patterns, we found that five ($167\%$) of thirty clones produced an extended-spectrum $\beta$-lactamase. 837- and 259-bp fragments specific to bla$_{TEM}$ genes were amplified, as determined by banding patterns of PCR amplification with designed primers. TEM­1 was the most prevalent $\beta$-lactamase and conferred resistance to ampicillin, piperacillin, and cephalothin. TEM-116 had a spectrum that was extended to ceftazidime, cefotaxime, and aztreonam. The resistance levels conferred by the pre-antibiotic era alleles of TEM-type $\beta$-lactamases were essentially the same as the resistance levels conferred by the TEM-type alleles which had been isolated from clinically resistant strains of bacteria of the antibiotic era. Our first report on TEM-type $\beta$-lactamases of the pre-antibiotic era indicates that TEM-type $\beta$-lactamases paint a picture in which most of the diversity of the enzymes may not be the result of recent evolution, but that of ancient evolution.

• Journal of Microbiology and Biotechnology
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• 제27권11호
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• pp.1932-1941
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• 2017

DFC-2, a methyl 5-[2-(dimethylamino)ethoxy]-7,12-dioxo-7,12-dihydrodinaphtho[1,2-b:2',3'-d]furan-6-carboxylate, is reported to have antitubercular effects against Mycobacterium tuberculosis. At concentrations ranging from 0.19 to $0.39{\mu}g/ml$, DFC-2 inhibited both drugusceptible and -resistant strains of M. tuberculosis. Microarray analyses were employed to gain insights into the molecular mechanisms of DFC-2's action in M. tuberculosis. The most affected functional gene category was "lipid biosynthesis," which is involved in mycolic acid synthesis. The decrease in transcription of genes related to mycolic acid synthesis was confirmed by RT-PCR. Furthermore, we found that DFC-2 triggered a reduction in mycolic acid levels, showing a similar pattern to that of mycolic acid synthesis inhibitor isoniazid. These results may explain how this compound kills mycobacteria efficiently by inhibiting mycolic acid synthesis.

• 한국미생물·생명공학회지
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• 제14권3호
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• pp.265-270
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• 1986

L. casei세포의 유전연구를 위한 도구로서 세포융합 기술을 연구하였으며 융합세포(recombinant)를 선발하고 확인하기 위한 유전자 선발표지 인자로서 항생제 저항성이 이용되었고, 항생제 저항성 돌연변이 균주는 nitrosoguanidine을 처리하여 분리하였다. 선발 배지에서 항생제의 적절한 최종 농도는 streptomycin 25 $\mu\textrm{g}$/$m{\ell}$, hostacillin 0.5 I. U./$m{\ell}$, lincomycin 0.5$\mu\textrm{g}$/$m{\ell}$ 그리고 methicillin 5 $\mu\textrm{g}$/$m{\ell}$로 확인되었다. L. casei균주에서 높은 세포융합은 PEG 분자량 4,000에서 40%농도, 중성 부근의 pH, 3$0^{\circ}C$에서 약 1분간 처리하였을 때 얻어 졌다. 항생제 저항성의 자연돌연변이주의 출현빈도는 세포융합 출현빈도 보다 $10^2$-$10^3$ 정도 낮은 수준으로 나타났다. 세포융합 빈도는 모균에 대해 약 $10^{-4}$ 비율로 나타났다.

• 한국수산과학회지
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• 제52권6호
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• pp.605-616
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• 2019

This study evaluated the abundance and antimicrobial resistance of Escherichia coli in oysters Crassostrea gigas and major inland pollution sources sampled in 2014-2015 from Jaranman-Saryangdo on the southern coast of Korea. The ranges of the geometric mean of E. coli concentrations in oysters and major inland pollution sources were <20-29.8 MPN/100 g and 7.5-137.2 MPN/100 g, respectively. We isolated 247 strains of E. coli (28 from oysters and 247 from major inland pollution sources) and examined the antimicrobial resistance patterns of all isolates. Isolates from both sources were highly resistant to rifampin (99.5-100%) and cephalothin (70.8-78.6%). The resistance rate was higher in E. coli isolated from oysters those from inland pollution sources. Multiple resistance against at least four antimicrobials was observed in 85.7% and 21.0% of the oyster and major inland pollution sources isolates, respectively.