• Title/Summary/Keyword: resistant strains

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Relative Toxicity of Abamectin to the redatoryMite Amblyseius womersleyi Schicha (Acari: Phytoseiidae) and Twospotted Spider MIte Tetranychus urticae Koch (Acari: Tetranychidae) (아바멕틴의 긴털이리응애(Amblyseius womersleyi Schicha)와 점박이응애(Tetranychus urticae Koch)에 대한 선택독성)

  • Park, C.G.;Lee, M.H.;Yoo, J.K.;Lee, J.O.;Choi, B.R.
    • Korean journal of applied entomology
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    • v.34 no.4
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    • pp.360-367
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    • 1995
  • The relative toxicity of abamectin was assessed to the predatory mite Amblyseius womersleyi Schicha and to dicofol-resistant and -susceptible twospotted spider mite (TSM) Tetranychus urticae Koch in the laboratory. Abamectin was much les toxic to the predator than to the spider mite. At 0.12 and 0.6 ppm, all TSM adult females of the tow strains were killed within 48 h after dipping n the solutions. The lower concentrations (0.06 and 0.012 ppm) killed more than 77% of TSM female adults of the two strains at 120 h after treatment. However, abmectin did not significantly affect the survival and mobility of A. womersleyi female adults at a concentration of 0.12 ppm but the mortality was slightly increased up to 20~23% at 0.6 and 6 ppm. Abamectin did not significantly affect hatchability of one-day old TSM eggs at 0.06~0.6 ppm. The Four-day old eggs were much more susceptible to abamectin than one-day old eggs were. Within 0.006-6 ppm, abamectin did not affect the hatchability of A. womersleyi eggs and the development of resulting immature predators. When the predator female adults were dipped in 0.6 and 0.12 ppm solution, their reproduction was not affected, but at 6 ppm it was decreased by 35%. However, the reproduction of TSM reduced significantly at concentrations between 0.006 and 0.6 ppm. The differential toxicity of abamectin between TSM and the predator could be of practical importance in managing spider mite populations in the field. Abamectin at selective sublethal concentrations (i.e., 0.012~0.06 ppm) could be of value in adjusting predator/prey ratios in integrated management of spider mites.

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Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Antibacterial and Therapeutic Effects of Houttuynia cordata Ethanol extract for Murine Salmonellosis (어성초 ethanol 추출물의 마우스 살모넬라 감염증에 대한 항균 및 치료효과 규명)

  • Kim, Dong-Hyeok;Lim, Jung-Ju;Lee, Jin-Ju;Jung, Won-Chul;Shin, Hyeon-Jin;Lee, Hu-Jang;Kim, Gon-Sup;Kim, Suk
    • Korean Journal of Environmental Agriculture
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    • v.27 no.2
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    • pp.156-162
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    • 2008
  • Salmonellosis is a major bacterial zoonosis that causes a variety of disease syndromes, self-limited enteritis to fatal infection in animals and food-borne infection and typhoid fever in humans. Recently, the emergence of multidrug resistant strains of Salmonella spp. causes more serious problems in environment and public health. The present study was investigated the antibacterial effect of Houttuynia cordata ethanol extract(HCEE) for murine salmonellosis. In the cytotoxic effect of HCEE on RAW 264.7 cells, there was no detectable effect with any concentrations between 25 and 100 ${\mu}g/ml$ after 8 h incubation. The bacteriocidal effect of HCEE was not showed on a Salmonella enterica serovar Typhimurium(S. typhimurium). HCEE makes morphological change of the RAW 264.7 cells, and there was significant decreased bacterial uptake and intracellular replication within Salmonella infected cells. And further nitric oxide(NO) production of Salmonella infected RAW 264.7 cells with HCEE was decreased comparing to RAW 264.7 cells without HCEE until 8 h post infection. Oral administration of HCEE showed a therapeutic effect for S. typhimurium infected BALB/c mice. The mortality of HCEE treated mouse was 80% until 12 days, while that of HCEE untreated mouse was 100 % until 8 days after lethal dose of S. typhimurium infection. These data suggested that HCEE has a potency treatment for intracellular replicative pathogen including salmonellosis, brucellosis, tuberculosis, listeriosis etc., and the application of HCEE makes new strategies for safety medicine development without antibiotic resistance bacterial appearance and residue problem in food and solves the public health problem from antibiotic mis- and over use.

Symbiotic effectiveness and intrinsic antibiotic resistance of Rhizobium meliloti populated in Korean pasture soils (국내(國內) 초지토양(草地土壤)에 분포(分布)한 Rhizobium meliloti의 질소고정력(窒素固定力)과 항균제반응(抗菌劑反應) 특성(特性))

  • Kang, Ui-Gum;Ha, Ho-Sung;Jung, Yeun-Tae
    • Applied Biological Chemistry
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    • v.35 no.3
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    • pp.179-185
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    • 1992
  • Rhizobium meliloti populated in five Korean pasture soils were characterized by symbiotic effectiveness and intrinsic antibiotic resistance using whole-soil inoculum and 11 antibiotics, respectively. Most probable number (MPN) of naturalized rhizobia counted with alfalfa Vernal[Medicago sativa (L.)] as a host ranged $1.7{\times}10^2\;cells/g$. soil(Chunghyo, Kyeongiu)-$1.0{\times}10^5\;cells/g$. soil(Gampo, Kyeongiu) and ended to be positively associated with soil pH. On the whole, the effectiveness of population as compared to TAL mix inoculum (TAL 380+TAL 1372+TAL 1373) was very low. Nevertheless, there were two highly effective strains, YCK 539 and YCK 542, which were not inferior to TAL 1372, from Ogpo, Dalseong among the total of 30 of 6 isolates per each soil. As long as mean $N_2$ fixing ability of each soil isolate, the isolates from Hyeongog, Kyeonju were outstanding and the rest were in order of Ogpo, Dalseong>Chunghyo, Kyeongju>Hwaweon, Dalseong>Gampo, Kyeongiu. Isolates as a whole were resistant to erythromycin(67崙), nalidixic acid(77%), and streptomycin sulfate(8051), which had the concentration of $100\;{\mu}g/ml$, $160\;{\mu}g/ml$, and $10\;{\mu}g/ml$, respectively and divided into 14 patterns of resistance. Association between resistances in each soil was not clear. And there was no relationship of resistance pattern to effectiveness. The best effective strain YCKa 542 exclusively fell into No. X pattern having resistance to erythromycin, nalidixic acid, and neomycin sulfate.

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Strain Improvement of Leuconostoc mesenteroides as a Acid-Resistant Mutant and Effect on Kimchi fermentation as a Starter (Leuconostoc mesenteroides의 내산성 변이주의 김치발효에 미치는 효과)

  • Kim Young-Hwan;Kim Hee-Zoong;Kim Ji-Young;Choi Tae-Bu;Kang Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.41-50
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    • 2005
  • An organic acid tolerance mutant (M-200) was obtained from Leuconostoc mesenteroides KCCM 35471, followed by the screening procedure using a specific organic acid medium (lactic acid: acetic acid, 2:1). The characteristics of the acid tolerance M-200 and the wild type LM-W were examined at various temperature and pH ranges $(l0-30^{\circ}C$ of temp, 3.5-4.5 of pH). The growth of strain M-200 at HCl adjusted medium $(10^{\circ}C\;and\;pH 3.5)$ was observed. In the case of organic acid adjusted medium, the strain showed its growth at the pH range of 3.8. When the strain M-200 was used as a starter for Kimchi fermentation, a constant acid level (0.55) was observed during the whole fermentation period. This result indicates that the strain produces a proper level of acid content for the Kimchi fermentation. This result also indicates that the edible period of Kimchi can be extended to 3.5 fold compare to the result obtained from the LM-W used Kimchi fermentation. However the excess use of the strain M-200 showed the inhibition of growth of Lactobacillus plantarum, low lactic acid level content and low level of organoleptic test. In the case of organic acid content during the Kimchi fermentation, the strain M-200 showed relatively low production rate compare to the wild type (M-200: 3.5 mg/L at 21 days of fermentation, LM-W: 7 mg/L at 21 days of fermentation). Therefore a mixed Kimchi starter containing M-200 and other strains probably maintain a good Kimchi quality during the fermentation.

Alternate Prey: A Mechanism for Elimination of Bacterial Species by Protozoa (원생동물(原生動物)에 의한 세균류(細菌類)의 소거(消去)에 관(關)한 기작(機作))

  • Mallory, Larry M.;Yuk, Chang-Soo;Liang, Li-Nuo;Alexander, Martin
    • Korean Journal of Soil Science and Fertilizer
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    • v.16 no.4
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    • pp.358-367
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    • 1983
  • Antibiotic-resistant strains of Salmonella typhimurium and Klebsiella pneumoniae died readily after their addition to raw sewage, but they grew in sterilized sewage. The decline was not a result of antibiotic stresses, and because the bacteria were able to survive in large numbers for at least 15 days in solutions containing no organic nutrients, it was not a result of competition. Toxin production, bacteriophages, and Bdellovibrio did not cause the disappearance of the two bacterial species. A decline was also evident if the sewage was first passed through a $3-{\mu}m$ filter or treated with cycloheximide or cycloheximide plus nystatin, but protozoa developed under these conditions. Little or no decline occurred if the sewage was filtered and treated with the eucaryotic inhibitors before adding S. typhimurium or K. pneumoniae, and protozoa were not detected. S. typhimurium increased in abundance if cycloheximide, streptomycin, and erythromycin or large amounts of glucose were added to sewage. Tetrahymena thermophilus did not significantly reduce the population of S. typhimurium in buffer when the density of the bacterium was about $10^4/ml$. However, when more than $10^8$ Enterobacter agglomerans cells per ml were added to the buffer, T. thermophilus reduced the abundance of E. agglomerans and S. typhimurium to $10^6$ and 10/ml, respectively. The density of S. typhimurium was further decreased by a second increment of E. agglomerans cells. The disappearance of S. typhimurium and K. pneumoniae from sewage thus is the result of predation by protozoa. It is proposed that predators will eliminate a prey species from a natural environment when an alternate prey is present at concentrations above the threshold number for active feeing by the predator and when the rate of growth of the prey is less than the rate of predation.

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A study of serotyping of Streptococcus pneumoniae by multibead assay (다중구슬 분석법에 의한 폐구균 혈청형 결정 연구)

  • Cho, Ky Young;Lee, Jung Ah;Cho, Sung Eun;Kim, Nam Hee;Lee, Jin A;Hong, Ki Sook;Lee, Hoan Jong;Kim, Kyung Hyo
    • Clinical and Experimental Pediatrics
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    • v.50 no.2
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    • pp.151-156
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    • 2007
  • Purpose : Streptococcus pneumoniae is a major etiologic agent for pneumonia, meningitis, otitis media, and sepsis among young children. Multi-drug resistant strains have raised great concern worldwide, thus the importance of prevention with vaccines has been emphasized. However, vaccines may force the appearance of pneumococcal infections by nonvaccine serotypes. Thus, distribution of pneumococcal serotypes should be monitored to estimate vaccine efficacy. We used a new and efficient multibead assay in determining pnemococcal serotypes. Methods : From January to February 2005, 643 children were recruited from ten day care centers to isolate pneumococci from their oropharynx. Pneumococcal serotyping was performed on 62 pneumococcal isolates from 60 children by multibead assay. This immunoassay required two sets of latex particles coated with pneumococcal polysaccharides and serotype-specific antibodies. Twenty four newly developed monoclonal antibodies specific for common serotypes and a pool of polyclonal rabbit sera for some of the less common serotypes were used. Results : The most prevalent pneumococcal serotypes were serotype 6A, 19A, 19F, 23F, and 11A/D/F which accounted more than 50 precent of all the 62 pneumococcal isolates. We found that multibead assay can be performed very rapidly and objectively. Conclusion : This multibead immunoassay was very useful in serotyping clinical isolates of S. pneumoniae because it was simple, reliable and fast.

Establishment of PCR Conditions for the Identification of Stenotrophomonas maltophilia Isolated from Boar Semen and Antimicrobial Susceptibility Patterns of the Isolates (돼지 정액에서 분리된 Stenotrophomonas maltophilia 확인을 위한 PCR 기법 개발 및 분리 균주의 항생제 감수성 양상)

  • Jung, Byeong-Yeal;Park, Bum-Soo;Kim, Ha-Young;Byun, Jae-Won;Kim, Ae-Ran;Jeon, Albert Byung-Yun;Kim, In-Cheul;Chung, Ki-Hwa
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1114-1119
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    • 2012
  • Bacteria are frequently contaminated during the collection and processing procedures of boar semen. Of the contaminants, Stenotrophomonas (S.) maltophilia is a Gram-negative bacterium that is widely distributed in a variety of habitats. Although PCR assays have been developed for the detection of S. maltophilia, they cross-react with some species of Xanthomonas. In this study, we designed a primer set for the detection of S. maltophilia in order to target the chiA (GenBank accession no. NC_010943) gene. The specific PCR products were amplified from S. maltophilia only, not from other tested strains that are frequently found in semen. The detection limit of the PCR was $1.5{\times}10^3$ CFU/ml with pure-cultured S. maltophilia and $1.5{\times}10^4$ CFU/ml with S. maltophilia spiked in semen. Twenty-six (5.9%) S. maltophilia were isolated from 440 semen samples. The PCR results exhibited 98.9% agreement with a comparison of S. maltophilia isolation. Also, the sensitivity and specificity of the PCR were 100% and 98.7%, respectively. In the antimicrobial susceptibility test, S. maltophilia isolates were highly susceptible to enrofloxacin and florfenicol, while the majority of them were resistant to amoxicillin/clavulanic acid, apramycin, ceftiofur, penicillin, and spectinomycin. These results indicated that the PCR using the chiA gene was proven to be reliable and effective for the detection of S. maltophilia with high levels of sensitivity and specificity.

Studies on the Function of the Rv2435c Gene of the Mycobacterium bovis BCG (Mycobacterium bovis BCG Rv2435c 유전자의 기능에 대한 연구)

  • Lee Seung-Sil;Bae Young-Min
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.415-422
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    • 2005
  • library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

Antibacterial activity of new quinolones against Edwardsiella tarda isolated from eel tanks (양만장에서 분리된 Edwardseilla tarda균에 대한 new quinolone계 약물의 항균작용)

  • Choi, Min-Soon;Kim, Kyong-Ho;Seo, Young-Ho;Choi, Sang-Hoon;Park, Kwan-Ha
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.185-193
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    • 1996
  • Extensive acquirement of drug resistance to traditional antibacterial agents poses a serious problem to eel aquaculturists. To collect the basic information for new drug development in the future, we assessed the in vitro antibacterial efficacy of 14 new quinolones with 75 isolates of Edwardsiella tarda from local aquaculture tanks of Anguilla japonica. Of all tested quinolones under development or marketed for human use, DU-6859 was most potent with its $MIC_{50}$ value of $0.05{\mu}g$/ml in broth microdilution assay. The drugs whose $MIC_{50}$ values ranged from 0.2 to $0.78{\mu}g$/ml were T-3762, Bay-y3118, ciprofloxacin, norfloxacin, ofloxcin and tosufloxacin. The weakest group of drugs, with their $MIC_{50}$ being 1.56-$3.13{\mu}g$/ml, were difloxacin, sparfloxacin, fleroxacin, Q-35, amifloxacin, lomefloxacin and enoxacin. The number of resistant strains, when arbitrarily defined with their MICs of $\geq6.25{\mu}g$/ml, was : 3 to T-3762, 3 to Bay-y3118, 44 to difloxacin, 16 to sparfloxacin, 13 to ciprofloxacin, 19 to fleroxacin, 36 to Q-35). 31 to amifloxacin, 5 to norfloxacin, 13 to ofloxacin, 31 to lomefloxacin, 41 to enoxacin, 12 to tosufloxacin and 0% to DU-6859, respectively. This information can be taken into consideration for the future development of fisheries antibacterial quinolones.

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