• Archives of Pharmacal Research
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• 제19권5호
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• pp.353-358
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• 1996

E. coli 11 and E. coli 101, clinical isolates of Escherichia coli were resistant to various quinolones, especially MICs to norfloxacin of both strains were higher than 100 mg/ml. In the presence of carbonyl cyanide m-chlorophenylhydrazone, a proton gradient uncoupler, norfloxacin uptake in both strains was increased, suggesting that an efflux system play an important role in the norfloxacin resistance. Outer membrane proteins of the susceptible and resistant strains which could affect the route of norfloxacin entry into cells were different. When quinolone resistance determining region(QRDR) of gyrA was amplified using PCR and cut with Hinf I, QRDR in the susceptible strain yielded two fragments while QRDRs in E. coli 11 and E. coli 101 yielded only one uncut fragment. When DNA sequence of QRDR was analyzed, there were two mutations as Ser-83 and Asp-87 in both resistant strains. these residues were changed to Leu-83 and Asn-87, respectively. These results showed that the norfloxacin resistance of E. coli 11 and E. coli 101 was resulted from multiple changes-an altered DNA gyrase A subunit, a change in route of drug entry, and reduction in quinolone concentration inside cells due to an efflux system.

• 대한수의학회지
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• 제59권2호
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• pp.75-80
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• 2019

Enrofloxacin, a fluoroquinolone, is a broad-spectrum antibiotic widely used in veterinary medicine that inhibits the action of bacterial DNA gyrase, resulting in anti-bacterial effects. This study was performed to examine whether enrofloxacin has modulatory and anti-inflammatory activity on immune cells. A few studies have reported the anti-inflammatory effects of enrofloxacin. In this study, we used mouse spleen cells treated with lipopolysaccharide (LPS) and examined the effects of enrofloxacin. Several assays were performed in LPS-treated spleen cells after the enrofloxacin treatment. Enrofloxacin inhibited the metabolic activity and mitochondrial membrane potential of LPS-treated spleen cells significantly. On the other hand, enrofloxacin did not alter the proportion of the subsets in spleen cells, and did not induce cell death. The production of tumor necrosis factor-alpha in LPS-treated spleen cells was inhibited by enrofloxacin. Overall, enrofloxacin had modulatory activity in spleen cells treated with LPS. These data may broaden the use of enrofloxacin as an antibiotic with anti-inflammatory activity in veterinary clinics.

• Journal of Microbiology
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• 제44권6호
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• pp.591-599
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• 2006

To address the diversity of cyclodextrin-producing P. graminis strains isolated from wheat roots and rhizospheres of maize and sorghum sown in Australia, Brazil, and France, restriction fragment length polymorphism analysis of part of genes encoding RNA polymerase (rpoB-RFLP) and DNA gyrase subunit B (gyrB-RFLP) was used to produce genetic fingerprints. A phylogenetic tree based on rpoB gene sequences was also constructed. The isolates originated from Brazil could be separated from those from Australia and France, when data from the rpoB-based phylogenetic tree or gyrB-RFLP were considered. These analyses also allowed the separation of all P. graminis strains studied here into four clusters; one group formed by the strains GJK201 and $RSA19^T$, second group formed by the strains MC22.02 and MC04.21, third group formed by the strains TOD61, TOD 221, TOD302, and TOD111, and forth group formed by all strains isolated from plants sown in Cerrado soil, Brazil. As this last group was formed by strains isolated from sorghum and maize sown in the same soil (Cerrado) in Brazil, our results suggest that the diversity of these P. graminis strains is more affected by the soil type than the plant from where they have been isolated.

• 대한수의학회지
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• 제54권2호
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• pp.91-99
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• 2014

An antimicrobial susceptibility test was conducted to compare the resistance rates among Campylobacter spp. isolates from dogs (n = 50) raised under diverse conditions and humans (n = 50). More than 60% of Campylobacter (C.) jejuni from dogs and humans showed resistance to nalidixic acid, enrofloxacin and ciprofloxacin. C. jejuni isolates from humans showed higher resistance to tetracycline (83.3%) and ampicillin (91.3%) than those from dogs. None of the C. jejuni or Campylobacter coli isolates from humans or dogs were resistant to erythromycin. Overall, 85% of Campylobacter spp. isolates showed a multidrug resistant phenotype. Nucleotide sequencing analysis of the gryA gene showed that 100% of $NA^R/CIP^R$ C. jejuni isolates from dogs and humans had the Thr-$86^{th}$-Ile mutation, which is associated with fluoroquinolone resistance. flaA PCR restriction fragment length polymorphism (RFLP) typing to differentiate the isolates below the species level revealed 12 different clusters out of 73 strains. The human isolates belonged to eight different RFLP clusters, while five clusters contained dog and human isolates.

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.650-655
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• 1998

The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.

• Journal of Pharmaceutical Investigation
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• 제39권6호
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• pp.431-435
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• 2009

The comparative superior in vitro activity of DW-224a was supported by the downward MIC distribution due to the weakened influence of alterations within target enzymes in ciprofloxacin-resistant Staphylococcus aureus. The MI$C_{50}$ for DW-224a was 4 $\mu$g/mL, similar to that of gemifloxacin, 8-fold less than that of sparfloxacin and 16-over-fold less than that of ciprofloxacin. We constructed combinations of amino acid changes, located at codon 80, 83 or 84 within GrlA and 84, 85 or 88 within GyrA, which were associated with MIC increase. The amino acid changes were less influential to the MIC of DW-224a compared to those of other fluoroquinolones, and it was verified from the requirement of a total of two GrlA- and two GyrA-alterations to reach the MIC of DW-224a over 32 $\mu$g/mL.

• Mycobiology
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• 제41권4호
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• pp.234-242
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• 2013

A total of 62 bacterial isolates were obtained from Gomsohang mud flat, Mohang mud flat, and Jeju Island, Republic of Korea. Among them, the isolate CNU114001 showed significant antagonistic activity against pathogenic fungi by dual culture method. The isolate CNU114001 was identified as Bacillus amyloliquefaciens by morphological observation and molecular data analysis, including 16SrDNA and gyraseA (gyrA) gene sequences. Antifungal substances of the isolate were extracted and purified by silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. The heat and UV ray stable compound was identified as iturin, a lipopeptide (LP). The isolate CNU114001 showed broad spectrum activity against 12 phytopathogenic fungi by dual culture method. The semi purified compound significantly inhibits the mycelial growth of pathogenic fungi (Alternaria panax, Botrytis cinera, Colletotrichum orbiculare, Penicillium digitatum, Pyricularia grisea and Sclerotinia sclerotiorum) at 200 ppm concentration. Spore germ tube elongation of Botrytis cinerea was inhibited by culture filtrate of the isolate. Crude antifungal substance showed antagonistic activity against cucumber scleotiorum rot in laboratory, and showed antagonistic activity against tomato gray mold, cucumber, and pumpkin powdery mildew in greenhouse condition.

• 한국환경과학회지
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• 제29권5호
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• pp.435-443
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• 2020

The purpose of this study was to investigate eco-friendly measures to manage major diseases which cause heavy economic damages to ginseng. Morphological, physicochemical, and molecular biological species identification was carried out after isolating useful antagonistic bacteria from ginseng fields. In addition, optimal conditions for mass culture were established, and he efficacy of the bacteria in the prevention of the diseases was verified in the field. The results showed that about 150 bacteria were extracted from 150 ginseng fields in the whole county. Among them, B. pyrrocinia LA101 was finally selected, which had a strong antagonistic potency against Alternaria panax, Botrytis cinerea, Rhizoctonia solani, and Cylindrocarpon destructans on agar media. The B. pyrrocinia LA101 is a baculiform gram-negative bacterium identified as Burkholderia pyrrocinia according to results from an API(Analytical Profile Index) kit, 16S rRNA, and gyrase gene sequencing analysis. It was donated to the microbe bank of the Agricultural Genetic Resources Center at the National Academy of Agriculture Science under the Rural Development Administration on September 28, 2011 (Donation No. KACC91663P). A patent for the mass culture technology was granted in August 2012 (Patent No. 10-1175532).

• 한국동물위생학회지
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• 제42권3호
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• pp.153-159
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• 2019

The objective of this study was to identify mutations in the quinolone resistance determining region (QRDR) of the gyrA, gyrB, parC and parE genes, and the presence of plasmid-mediated quinolone resistance (PMQR) genes: qnrA, qnrB, qnrS, aac(6')-lb-cr and qepA in 40 nalidixic acid- resistant ($NA^R$) Salmonella isolates isolated from poultry slaughterhouse. The MIC of NA and ciprofloxacin for 40 $NA^R$ Salmonella isolates was $128{\sim}512{\mu}g/mL$ and < $0.125{\sim}0.25{\mu}g/mL$, respectively. The Salmonella isolates were resistant to NA (100%), gentamicin (5.0%) and ampicillin (2.5%). All $NA^R$ Salmonella isolates represented point mutation in codons Aspartic acid(Asp)-87 (90%) and Serine(Ser)-83 (10%) of QRDR of gyrA gene: $Asp87{\rightarrow}glycine$, $Ser83{\rightarrow}tyrosine$. No mutations were observed in QRDR of the gyrB, parC and parE gene. Moreover PMQR genes was not found in any of the tested isolates. Our findings showed that DNA gyrase is the primary target of quinolone resistance and a single mutation in codon Asp87 and Ser83 of the gyrA gene can confer resistance to NA and reduced susceptibility ciprofloxacin in Salmonella isolates.

• 농업과학연구
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• 제47권4호
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• pp.1011-1020
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• 2020

Efficient management systems of facilities make it possible to manage environmental conditions properly, such as the temperature, humidity and light source required for the best growth of the crops, as well as for the mass production of fruit and vegetables with high quality every year through an advanced and protected cultivation system. Powdery mildew is a type of chronic disease that is difficult to control during the production of Korean melons under a protected cultivation system, the use of which is increasing in production areas in Korea. Two Bacillus strains isolated from soil samples showed antagonistic activities against several pathogens, specifically Botrytis cinerea, Colletotrichum gloeosporioides, and Fusarium oxysporum f.sp. melonis; they were identified as Bacillus velezensis M2 and B. amyloliquefaciens M3 in a molecular biological test of the nucleotide sequences of gyrase subunit A (gyrA). The treatment was given three times at intervals of five days with 400-fold diluted cultures of B. velezensis M2 and B. amyloliquefaciens M3. This led to the inhibition of the incidence of powdery mildew disease in Korean melon leaves, which resulted in effective control efficiency against the incidence of powdery mildew disease with control values of 87% and 65%, respectively. Cultures of antagonistic microbes tested in this study can be used to increase the efficiency as part of an environmentally friendly management scheme to prevent powdery mildew disease during the protected cultivation of crops, including Korean melons.