• Title/Summary/Keyword: high-level resistant

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Development of Potato Virus Y Resistant Tobacco Plant by Transformation of the Untranslatable Viral Coat Protein Encoding cDNA (감자 바이러스 Y 비전이성 외피단백질 cDNA의 형질전환에 의한 바이러스 저항성 연초품종 개발)

  • 이청호;이영기;강신웅;박성원;김상석;박은경
    • Journal of the Korean Society of Tobacco Science
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    • v.19 no.2
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    • pp.117-123
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    • 1997
  • Viral coat protein (CP) encoding cDNA with artificial start and stop codons was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from the Korean isolate of potato virus Y-vein nectrosis strain (pVY-VN). To make PVY CP cDNA to untranslatable form, three stop codons were inserted near the start codon by "megaprimer-PCR" method. The untranslatable CP cDNA was subcloned to plant expression vector and transferred to N. tabacum cv. NC82 by Agrobacterium-mediated transformation. Highly resistant plants to PVY infection were screened, based on symptom development after mechanical virus inoculation. By genomic PCR and Southern blot analysis, one or more copies of the untranslatable CP gene were found in all transformants. From northern blot analysis, highly resistant transgenic lines had very low level of CP transcript but susceptible lines had high level, suggesting resistance to PVY infection should be related to RNA-mediated mechanism.mechanism.

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Development of multiplex PCR for detection of vancomycin resistant enterococci(VRE) and epidemiological application in Korea

  • Seo, Keun-seok;Song, Deok-jln;Gwyther, M.M.;Park, Yong-ho
    • Korean Journal of Veterinary Research
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    • v.39 no.2
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    • pp.343-352
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    • 1999
  • Vancomycin resistant enterococci (VRE) have emerged as an important nosocomial pathogen. Since 1989 the Center for Disease Control, United States, has reported a rapid increase in the incidence of enterococcal bacteremia and endocarditis infection by VRE. It was suggested that the use of avoparcin was associated with the appearance of VRE in animal husbandry. To date, several detection methods have been used based on conventional methods of culture and gene detection. However, these methods have some limitations such as time-consuming, laborious and additional differential needs. Therefore, In this study a multiplex PCR method was established to detect and differentiate resistance types of enterococci which specifically amplify the four van genes encoding vancomycin resistance elements. Using the method, we investigated the incidence rates and types of VRE from farms using or not using avoparcin. A total of 1091 animal fecal samples were collected from 70 pig and 32 poultry farms. A total of 425 of enterococci were isolated from samples. Of the 425 isolates, 11 of the them showed a pattern of high-level vancomycin resistance (MIC : $64{\sim}256{\mu}g/ml$) which was associated with the presence of the vanA or vanB gene. Fifty-seven isolates showed a pattern of low-level vancomycin resistance (MIC : $3{\sim}8{\mu}g/ml$) associated with the vanC-1 or vanC-2 gene. Interestingly, all isolates with high-level vancomycin resistance were from farms that have never used avoparcin. Moreover, the high-level VRE isolation rate in Korea (2.58%) was much lower than that of other countries (50% in England, 7% in Belgium) where avoparcin have been used. In conclusion, the multiplex PCR method established in this study could be applied for detection of VRE.

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Isolation of a High-Yield Mutant Strain for L-Proline Production and Its Fermentation Conditions

  • Ryu, Wuk-Sang;Jang, Hyung-Wook;Cho, Kyoung-Hee;Chang, Soon-Jae;Ryu, Yeon-Woo;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.613-618
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    • 1999
  • L-Proline-producing mutant strains were developed by exposing L-glutamic acid-producing bacteria to N-metyl-N-nitro-nitrosoguanidine and UV irradiation. A L-histidine auxotroph of Corynebacterium acetoacidophilum RYU3161(KCTC 0616BP), which was resistant to sulfaguanidine and proline analogs (DHP, AZC, TAC), was isolated. The activity of the mutant strain's $\gamma$-glutamyl kinase was 45% higher than that of the parent strain. The optimum level of L-histidine for production of L-proline was 0.16 g/l. In a 5-1 jar fermenter, the mutant strain produced L-proline at a high concentration (35 g/l) level within 48 h of cultivation.

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Distribution of Pseudomonas-Derived Cephalosporinase and Metallo-β-Lactamases in Carbapenem-Resistant Pseudomonas aeruginosa Isolates from Korea

  • Cho, Hye Hyun;Kwon, Gye Cheol;Kim, Semi;Koo, Sun Hoe
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1154-1162
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    • 2015
  • The emergence of carbapenem resistance among Pseudomonas aeruginosa is an increasing problem in many parts of the world. In particular, metallo-$\beta$-lactamases (MBLs) and AmpC $\beta$lactamases are responsible for high-level resistance to carbapenem and cephalosporin. We studied the diversity and frequency of $\beta$-lactamases and characterized chromosomal AmpC $\beta$lactamase from carbapenem-resistant P. aeruginosa isolates. Sixty-one carbapenem-resistant P. aeruginosa isolates were collected from patients in a tertiary hospital in Daejeon, Korea, from January 2011 to June 2014. Minimum inhibitory concentrations (MICs) of four antimicrobial agents were determined using the agar-dilution method. Polymerase chain reaction and sequencing were used to identify the various $\beta$-lactamase genes, class 1 integrons, and chromosomally encoded and plasmid-mediated ampC genes. In addition, the epidemiological relationship was investigated by multilocus sequence typing. Among 61 carbapenem-resistant P. aeruginosa isolates, 25 isolates (41.0%) were MBL producers. Additionally, 30 isolates producing PDC (Pseudomonas-derived cephalosporinase)-2 were highly resistant to ceftazidime (MIC50 = $256{\mu}g/ml$) and cefepime (MIC50 = $256{\mu}g/ml$). Of all the PDC variants, 25 isolates harboring MBL genes showed high levels of cephalosporin and carbapenem resistance, whereas 36 isolates that did not harbor MBL genes revealed relatively low-level resistance (ceftazidime, p < 0.001; cefepime, p < 0.001; imipenem, p = 0.003; meropenem, p < 0.001). The coexistence of MBLs and AmpC $\beta$-lactamases suggests that these may be important contributing factors for cephalosporin and carbapenem resistance. Therefore, efficient detection and intervention to control drug resistance are necessary to prevent the emergence of P. aeruginosa possessing this combination of $\beta$-lactamases.

Prevalence and molecular characteristics of 16s rRNA methylase gene rmtB in amikacin resistant Escherichia coli isolated from South Korea

  • Belaynehe, Kuastros Mekonnen;Won, Ho Geun;Yoon, In Joong;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.59 no.3
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    • pp.157-160
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    • 2019
  • The production of rmtB-encoded 16S rRNA methylases has emerged as a novel mechanism promoting high-level resistance toward aminoglycosides in Gram-negative bacteria. Between 2015 and 2017, 636 distinct commensal Escherichia (E.) coli isolates were collected from different farms in South Korea to determine the prevalence and molecular characteristics of rmtB. The positive rates of rmtB between all the isolates and amikacin-resistant isolates were 1.1 and 100%, respectively. High-level aminoglycoside resistance could be transferred by conjugation from rmtB-positive donors to higher amikacin-resistance efficacies. This is the first report of 16S rRNA methylase-encoding genes in E. coli isolated from food-producing animals in Korea.

Comparative risks of resistant microorganisms in the intestinal track of imported freshwater ornamental fish and cultured marine fish (수입산 담수관상어 및 양식 해산어의 장내세균에서 나타나는 내성균 위험성 비교)

  • Yoon, So-Hye;Jun, Lyu-Jin;Kim, Young-Jin;Jin, Ji-Woong;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.25 no.2
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    • pp.77-84
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    • 2012
  • Various antibiotics, that could induce the appearance of resistant microorganisms, have been used for treatment or prevention of bacterial diseases in marine and ornamental fish. We determined and characterized the level of antibiotic-resistant bacteria and proportion of multi-drug resistant bacteria in intestinal microflora of both marine fish cultured in Korea and imported ornamental freshwater fish. For this the bacterial species and resistance to antibiotics were investigated in intestine of rock bream Oplegnathus fasciatus cultured in Korea and pearl gourami Trichogaster leeri imported from Singapore to characterise. Although the bacterial species were different, proportions of resistant bacteria to single antibiotics or multi-drug were higher in intestinal microflora of pearl gourami Trichogaster leeri imported from Singapore than in rock bream Oplegnathus fasciatus cultured in Korea. These results indicate that various antibiotics have been being used before trading without measures in the market of asian ornamental fishes, providing high risks for the emergence of multi-drug resistant bacteria.

Evaluation of Molecular Weight Distribution, Pasting and Functional Properties, and Enzyme Resistant Starch Content of Acid-modified Corn Starches

  • Koksel, Hamit;Ozturk, Serpil;Kahraman, Kevser;Basman, Arzu;Ozbas, Ozen Ozboy;Ryu, Gi-Hyung
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.755-760
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    • 2008
  • The aim of this study was to produce resistant starch preparations from acid-modified com starches prepared at various hydrolysis levels (0.5-4.0 hr). Effect of autoclaving cycles on resistant starch (RS) formation was investigated. Molecular weight distribution, pasting and functional properties of acid-modified com starches were determined. For RS formation native and acid-modified starch samples were gelatinized and autoclaved (1 or 2 cycles). While native and acid-modified starches did not contain any RS, the levels increased to 9.0-13.5% as a result of storage at $95^{\circ}C$ after first autoclaving cycle. Second autoclaving cycle together with storage at $95^{\circ}C$ brought final RS contents of the samples incubated at 4 and $95^{\circ}C$ after the first cycle to comparable level. As acid modification level increased, the amount of high molecular weight fractions decreased, resulting in significant decreases in viscosities (p<0.05). The samples produced in this study had low emulsion stability and capacity values.

Development of an Earthquake-Resistant Model for a High-Level Waste Disposal Canister (고준위 폐기물 처분용기 내진 해석 모델 개발)

  • Choi, Young-Chul;Yoon, Chan-Hoon;Kim, Hyun-Ah;Choi, Heui-Joo
    • Tunnel and Underground Space
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    • v.24 no.4
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    • pp.316-324
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    • 2014
  • In the underground 500 m depth, the high level radioactive waste disposal system is made by boring the tunnel in the base rock and putting the high level waste disposal canister that is the surrounding form with the buffer material. According to the many statistics, it is the tendency that the earthquake increases in the Korean peninsula every year. In case that the earthquake is generated, the disposal canister in the rock mass can be broken due to the shearing force in the underground. Furthermore, a major environmental problems can be caused by the radioactive harmful substances. In this study, the earthquake-proof type buffer material was developed with the protection method safely on the earthquake. The main parameter having an effect on the earthquake-resistant performance was analyzed and the earthquake-proof type buffer material was designed. The shear analysis model was developed and the performance of the earthquake-proof type buffer material was evaluated by using ABAQUS.

Resistance Determinants and Antimicrobial Susceptibilities of Mupirocin-Resistant Staphylococci Isolated from a Korean Hospital (국내 한 대학병원에서 수집된 Mupirocin 내성 포도알균의 내성 유전자 및 항생물질 감수성 분석)

  • Min, Yu-Hong;Lee, Jong-Seo;Kwon, Ae-Ran;Shim, Mi-Ja;Choi, Eung-Chil
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.93-101
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    • 2012
  • We analyzed mupirocin resistance rates among staphylococcal isolates collected from a Korean hospital in 2003 (100 isolates), 2005 (195 isolates), 2006 (151 isolates), and 2009 (112 isolates). In Staphylococcus aureus, rates of high-level mupirocin resistance (MIC, minimal inhibitory concentration ${\geq}512{\mu}g/ml$) decreased and did not appear since 2005. In contrast, low-level mupirocin resistance (MIC $8-256{\mu}g/ml$) was not detected in 2003 and 2005 but its rates later increased to 6.9% in 2009. Total resistance rates of coagulase-negative staphylococci (CNS) were significantly higher than those of S. aureus. The rates of high-level resistance of CNS increased from 16.0% in 2003 to 31.5% in 2009. The rate of low-level resistance of CNS was 8.0% in 2003 and around 11% later. In all high-level resistant isolates, the ileS-2 gene was detected. All low-level resistant isolates contained the known V588F mutation in ileS gene. Previously unknown mutations such as V458G in S. aureus and D172A, Y490H and I750V in CNS were identified additionally. One S. aureus isolate with high-level resistance was resistant to oxacillin and several topical antibiotics commonly used for the treatment of skin infection. Ten S. aureus isolates with low-level resistance were also resistant to all of these antibiotics except fusidic acid. CNS isolates with high-level (61 isolates) and low-level resistance (27 isolates) exhibited significantly higher resistance rates to these antibiotics than mupirocin-susceptible CNS isolates (167 isolates). In conclusion, prevention of the emergence of mupirocin resistance is necessary for the effective treatment of skin infection by staphylococci.

Immunophenotype Characterization for Swine Selected Line, Which is Resistant for the Mycoplasma Pneumonia

  • Katayama, Masafumi;Fukuda, Tomokazu;Okamuara, Toshihiro;Suda, Yoshihito;Suzuki, Eisaku;Uenishi, Hirohide;Suzuki, Keiichi
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.889-897
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    • 2011
  • Mycoplasma Pneumonia of swine (MPS) decreases the daily growth of pigs, and, co-infection with a virus sometimes causes severe pneumonia. Genetic selection of pigs resistant to the pulmonary MPS lesion might solve the economic loss due to MPS in animal production. Here, we examined the immunophenotype of Landrace line (Miyagino L2), genetically selected to reduce the incidence of pulmonary MPS lesion for 5 generations in Miyagi Prefecture Animal Industry Experiment Station. Although this line is expected to be resistant to the pulmonary MPS lesion, the biological characteristics of its immune function are not clear. We investigated details of the immunorelated phenotype of Miyagino L2 at the hematological and molecular biological level, including cytokine expression, and compared the results with that of non-genetically selected Landrace. Miyagino L2 showed decreased antigen-specific IgG and IgM production and increased CD8-positive T-cell population, and high levels of cortisol concentration, suggesting that the MPS-resistant phenotype is associated these immunological differences. Additionally, T-cell CD4 expression was highly correlated with the MPS expected breeding value. Although the detailed mechanisms underlying this high correlation remain unknown, our result suggested that the genetic selection of the expression level of CD4 might be useful to improve MPS resistance in pig production.