• Title/Summary/Keyword: acinetobacter

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Insight into Norfloxacin Resistance of Acinetobacter oleivorans DR1: Target Gene Mutation, Persister, and RNA-Seq Analyses

  • Kim, Jisun;Noh, Jaemin;Park, Woojun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1293-1303
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    • 2013
  • Antibiotic resistance of soilborne Acinetobacter species has been poorly explored. In this study, norfloxacin resistance of a soil bacterium, Acinetobacter oleivorans DR1, was investigated. The frequencies of mutant appearance of all tested non-pathogenic Acinetobacter strains were lower than those of pathogenic strains under minimum inhibitory concentration (MIC). When the quinolone-resistance-determining region of the gyrA gene was examined, only one mutant (His78Asn) out of 10 resistant variants had a mutation. Whole transcriptome analysis using a RNA-Seq demonstrated that genes involved in SOS response and DNA repair were significantly up-regulated by norfloxacin. Determining the MICs of survival cells after norfloxacin treatment confirmed some of those cells were indeed persister cells. Ten colonies, randomly selected from among those that survived in the presence of norfloxacin, did not exhibit increased MIC. Thus, both the low mutation frequency of the target gene and SOS response under norfloxacin suggested that persister formation might contribute to the resistance of DR1 against norfloxacin. The persister frequency increased without a change in MIC when stationary phase cells, low growth rates conditions, and growth-deficient dnaJ mutant were used. Taken together, our comprehensive approach, which included mutational analysis of the target gene, persister formation assays, and RNA sequencing, indicated that DR1 survival when exposed to norfloxacin is related not only to target gene mutation but also to persister formation, possibly through up-regulation of the SOS response and DNA repair genes.

Acinetobacter Isolates Growing with Carbon Monoxide (일산화탄소를 이용하여 성장하는 acinetobacter의 분리 및 동정)

  • 조진원;임현숙;김영민
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.1-8
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    • 1985
  • Three strains (JC1, JC2, and HY1) of aerobic carbon monoxide (CO)-utilizing Acinetobacter were isolated from soil through CO-enrichment culture technique. All of them were Gram-negative, nonmotile, and rod-shated but they were changed to spherical form at the end of logarithmic phase. They were resistant to penicillin and able to frow at $42^{\circ}C$. The guanine plus cytosine contents of the DNAs ranged from 43 to 44.5 mol%. Oxidase was not present in all cells. The colonies were smooth and whitish yellow. Heterotrophic growth occurred on several sugars, organic acids, amino acids, and alcohols. The doubling times under and atmosphere of 30% CO and 70% air at $30^{\circ}C$ were 19h, 25h, and 35h, respectively, for JC1, JC2, and HY1, JC1 was studied in more detail. The cells were grown optimally in a mineral medium (pH 6.8) under a gas mixture of 30% CO and 70% air at $30^{\circ}C$. Growth of the cells with CO did not depend on molybdenum. It was able to grow with 100 ppm of CO in air as a sole source of carbon and energy.

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Effects of Psychrotrophic Bacteria, Serratia liquefaciens and Acinetobacter genomospecies 10 on Yogurt Quality

  • Shin, Yong Kook;Oh, Nam Su;Lee, Hyun Ah;Choi, Jong-Woo;Nam, Myoung Soo
    • Food Science of Animal Resources
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    • v.34 no.4
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    • pp.543-551
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    • 2014
  • The aim of this study was to evaluate the effect of proteolytic (Serratia liquefaciens, match %: 99.39) or lipolytic (Acinetobacter genomospecies 10, match %: 99.90) psychrotrophic bacteria (bacterial counts, analysis of free fatty acids (FFA) and analysis of free amino acids) on the microbial and chemical properties (yogurt composition), and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of yogurt during storage. Yogurts were prepared with raw milk preinoculated with each psychrotrophic bacteria. The total solid, fat, and protein content were not affected by preinoculation, but the pH of yogurt preinoculated with psychrotrophic bacteria was higher than in control. There was a dramatic increase in short chain free fatty acids among FFA in yogurt with Acinetobacter genomospecies 10. For 14 d of cold storage condition, SCFFA was 25.3 mg/kg to 34.4 mg/kg (1.36 times increased), MCFFA was 20.4 mg/kg to 25.7 mg/kg (1.26 times increased), and LCFFA was 240.2 mg/kg to 322.8 mg/kg (1.34 times increased). Serratia liquefaciens (match %: 99.39) in yogurt caused a greater accumulation of free amino acids (FAA), especially bitter peptides such as leucine, valine, arginine, and tyrosine, but SDS-PAGE showed that the inoculation of Serratia liquefaciens did not affect the degree of casein degradation during storage. Taken together, the excessive peptides and FFA in yogurt generated from psychrotrophic bacteria could develop off-flavors that degrade the quality of commercial yogurt products.

Screening and Optimal Culture Conditions of Antibiotic-Producing Actinomycetes B-51 for Multidrug Resistant Acinetobacter baumannii (다제내성 Acinetobacter baumannii에 유효한 방선균 B-51의 탐색 및 이 균주가 생산하는 항생물질 발효 최적 배양 조건)

  • Rhee, Moon-Soo;Kim, Gwan-Pil;Bang, Byung-Ho
    • The Korean Journal of Food And Nutrition
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    • v.23 no.1
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    • pp.63-69
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    • 2010
  • With the increase of the use of antibiotics and invasive procedures, infections caused by multidrug-resistant Acinetobacter baumannii(MRAB) are increasing. We screened the antibiotic producing strain B-51 for antibacterial activity against MRAB from the soils and studied the effects of culture medium on the antibiotic production of B-51. The medium conditions for maximum antibiotic productivity of B-51 was 2% glycerol, 0.5% soybean meal, 0.01% $CaCl_2$, 0.01% $MgSO_4{\cdot}7H_2O$ and 0.01% $KH_2PO_4$ at an initial pH of 6.0, at $30^{\circ}C$ for 76 h.

Evaluation of EDTA-based Three Methods to Detect IMP-1 and VIM-2 Type Metallo-${\beta}$-Lactamase-Producing Clinical Isolates of Imipenem Resistant Acinetobacter and Pseudomonas spp.

  • Hong, Seung-Bok;Shin, Kyung-A;Hwang, Seock-Yeon
    • Biomedical Science Letters
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    • v.17 no.2
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    • pp.135-140
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    • 2011
  • We compared three EDTA-based phenotypic screening methods for detecting IMP-1 and VIM-2 type metallo-${\beta}$- lactamase (MBL)-producing isolates of Acinetobacter and Pseudomonas spp., EDTA-double disk synergy test (EDTADDST), Etest MBL, and imipenem (IPM)-EDTA disk test. A total of 183 isolates (65 Acinetobacter spp. and 118 Pseudomonas spp. showing IPM resistance), confirmed to MBL genes by PCR, were used. The criteria for MBL production were (i) presence of a synergistic zone between IPM and EDTA disks in EDTA-DDST, (ii) reduction of IPM minimal inhibitory concentration by ${\geq}$ 3 twofold dilutions in the presence of EDTA in the Etest MBL, and (iii) ${\geq}$ 7 mm increase in the inhibition zone around the IPM plus EDTA disks compared with a sole IPM disk in the IPM-EDTA disk test. In this study using 87 MBL-producing and 96 MBL-nonproducing isolates, the sensitivities/specificities of EDTA-DDST, Etest MBL and IPM-EDTA disk tests were 94.3/78.1%, 89.7/91.7%, and 97.7/95.8%, respectively. When the threshold for the increase of the inhibition zone around the IPM plus EDTA disk over a sole IPM disk was altered to ${\geq}$ 5 mm and ${\geq}$ 8 mm for Acinetobacter spp. and Pseudomonas spp., respectively, the sensitivity and specificity of the test were 98.9% and 96.9%, respectively. Of the three EDTA-based phenotypic tests, the IMP-EDTA disk test was superior for detection of MBL-producing isolates.

Isolation of an Acinetobacter junii SY-01 Strain Producing an Extracellular Lipase Enantioselectively Hydrolyzing Itraconazole Precursor, and Some Properties of the Lipase

  • Yoon, Moon-Young;Shin, Pyong-Kyun;Han, Ye-Sun;Lee, So-Ha;Park, Jung-Keug;Cheong, Chan-Seong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.97-104
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    • 2004
  • Water-sludge bacteria were screened to find a lipase enantioselectively hydrolyzing itraconazole precursor, which is well known as the starting material of antifungal drug agents. A bacterial strain was isolated and identified as Acinetobacter junii SY-01. After the strain was cultivated, the enzyme was purified 39.4-fold using ultrafiltration and gel filtration through a Sephadex G-100 chromatographic column and the activity yield was 34.9%. The molecular weight of the enzyme was about 40 kDa, as measured by SDS-PAGE, and the optimum pH was 7.0- 9.0 and stable at pH 6.0- 9.0. The optimum temperature was 45- $5^{\circ}C$, and 73% of the enzymes activity remained after incubation at 70% for 1 h. Enzyme activity was enhanced by gall powder, sodium deoxycholate, a cationic detergent Tween 80, and a non-ionic detergent Triton X-100, but was markedly inhibited by metal ions such as $Hg^{2+},Cu^{2+},Ni^{2+}/,Ca^{2+}$, and an anionic-surfactant sodium dodecylsulfate. The $K_{m}$ values for (R)- and (S)-enantiomers of the itraconazole precursor were 0.385 and 21.83 mM, respectively, and the $V_{max} values ($\mu$Mㆍmin^{-1}.)$ were 6.73 and 6.49, respectively. The acetyl group among the different acyl moieties of itraconazole precursor showed the highest enantioselectivity for the hydrolysis by the Acinetobacter junii SY-01 lipase, and the lipase from Acinetobacter junii SY-01 displayed better enantioselectivity than that of commercially available lipases and esterases.

Characterization of Trimethoprim-Sulfamethoxazole Resistance Genes and Their Relatedness to Class 1 Integron and Insertion Sequence Common Region in Gram-Negative Bacilli

  • Shin, Hae Won;Lim, Jinsook;Kim, Semi;Kim, Jimyung;Kwon, Gye Cheol;Koo, Sun Hoe
    • Journal of Microbiology and Biotechnology
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    • v.25 no.1
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    • pp.137-142
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    • 2015
  • Trimethoprim-sulfamethoxazole (TMP-SMX) has been used for the treatment of urinary tract infections, but increasing resistance to TMP-SMX has been reported. In this study, we analyzed TMP-SMX resistance genes and their relatedness with integrons and insertion sequence common regions (ISCRs) in uropathogenic gram-negative bacilli. Consecutive nonduplicate TMP-SMX nonsusceptible clinical isolates of E. coli, K. pneumoniae, Acinetobacter spp., and P. aeruginosa were collected from urine. The minimal inhibitory concentration was determined by Etest. TMP-SMX resistance genes (sul and dfr), integrons, and ISCRs were analyzed by PCR and sequencing. A total of 45 E. coli (37.8%), 15 K. pneumoniae (18.5%), 12 Acinetobacter spp. (70.6%), and 9 Pseudomonas aeruginosa (30.0%) isolates were found to be resistant to TMP-SMX. Their MICs were all over 640. In E. coli and K. pneumoniae, sul1 and dfr genes were highly prevalent in relation with integron1. The sul3 gene was detected in E. coli. However, in P. aeruginosa and Acinetobacter spp., only sul1 was prevalent in relation with class 1 integron; however, dfr was not detected and sul2 was less prevalent than in Enterobacteriaceae. ISCR1 and/or ISCR2 were detected in E. coli, K. pneumoniae, and Acinetobacter spp. but the relatedness with TMP-SMX resistance genes was not prominent. ISCR14 was detected in six isolates of E. coli. In conclusion, resistance mechanisms for TMP-SMX were different between Enterobacteriaceae and glucose non-fermenting gram-negative bacilli. Class 1 integron was widely disseminated in uropathogenic gram-negative baciili, so the adoption of prudent use of antimicrobial agents and the establishment of a surveillance system are needed.

Resazurin Reduction Time Test to Determine Post-pasteurization Contamination and Shelf Life of Market Milk (시유의 2차오염과 저장가능기간을 결정하기 위한 Resazurin 환원시간검사)

  • Choi, S,H.;Choi, J.J.;Lee, S.B.;Yoon, Y.H.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.999-1006
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    • 2004
  • The selective media including NPC agar, DHL agar, MacConkey agar, and Cetrimide desoxycholate agar were compared to determine selectivity for the growth of bacteria Cetrimide desoxycholate agar was better than NPC agar, DHL agar, and MacConkey agar for the growth of psychrotrophic grarn-negatve bacteria including Pseudomonas. and for the inhibition of gram positive bacteria The specificity of resazurin reduction time test was investigated to determine post-pasteurization contamination of market milk. Equal volume of Cetrimide desoxycholate broth was added to market milk, which was then incubated at $21^{\circ}C$ for 18 hours. The growth of bacteria in the incubated milk was detected in resazurin reduction time test. The results in resazurin reduction time test and total bacteria number count of market milk after storage at $7^{\circ}C$ were relatively correlated each other. Pseudomonas was isolated most frequently from the market milk stored at $7^{\circ}C$ for 10 days, and Acinetobacter and Aeromonas followed. Acinetobacter, Pseudomonas and Enterobacter were frequently isolated from the mixture of market milk and Cetrimide desoxycholate broth incubated at $21^{\circ}C$ for 18hours in resazurin reduction time test.

Biodegradation of diesel oil and n-alkanes (C18, C20, and C22) by a novel strain Acinetobacter sp. K-6 in unsaturated soil

  • Chaudhary, Dhiraj Kumar;Bajagain, Rishikesh;Jeong, Seung-Woo;Kim, Jaisoo
    • Environmental Engineering Research
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    • v.25 no.3
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    • pp.290-298
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    • 2020
  • A large residual fraction of aliphatic components of diesel prevails in soil, which has adverse effects on the environment. This study identified the most bio-recalcitrant aliphatic residual fraction of diesel through total petroleum-hydrocarbon fractional analysis. For this, the strain Acinetobacter sp. K-6 was isolated, identified, and characterized and investigated its ability to degrade diesel and n-alkanes (C18, C20, and C22). The removal efficiency was analysed after treatment with bacteria and nutrients in various soil microcosms. The fractional analysis of diesel degradation after treatment with the bacterial strains identified C18-C22 hydrocarbons as the most bio-recalcitrant aliphatic fraction of diesel oil. Acinetobacter sp. K-6 degraded 59.2% of diesel oil and 56.4% of C18-C22 hydrocarbons in the contaminated soil. The degradation efficiency was further improved using a combinatorial approach of biostimulation and bioaugmentation, which resulted in 76.7% and 73.7% higher degradation of diesel oil and C18-C22 hydrocarbons, respectively. The findings of this study suggest that the removal of mid-length, non-volatile hydrocarbons is affected by the population of bio-degraders and the nutrients used in the process of remediation. A combinatorial approach, including biostimulation and bioaugmentation, could be used to effectively remove large quantities of aliphatic hydrocarbons persisting for a longer period in the soil.

Antibiotic Resistant Patterns and DNA Fingerprint Analysis of Acinetobacter baumannii from Clinical Isolates (임상에서 분리된 Acinetobacter baumannii의 항생제 내성 패턴과 유전학적 특징)

  • Kim, Min-Ji;Lee, Si-Won;Lee, Do-Kyung;Park, Jae-Eun;Kang, Joo-Yeon;Park, Il-Ho;Shin, Hae-Soon;Ha, Nam-Joo
    • YAKHAK HOEJI
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    • v.57 no.2
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    • pp.132-138
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    • 2013
  • Acinetobacter baumannii is gram-negative bacilli that can be widely found in environments. Recently, A. baumannii emerged as a serious nosocomial infection. A total of 92 A. baumannii were isolated from hospitalized patients in Seoul, Korea, between December 2010 and April 2011. Antimicrobial susceptibility testing was investigated using CLSI agar dilution methods. Tigecycline non-susceptible A. baumannii isolates were investigated by repetitive extragenic palindromic sequence-based PCR (rep-PCR). Pulsed-field gel electrophoresis was performed to determine the epidemiological relationships. All clinical isolates showed high-level resistance to the most commonly used antibiotics: Ciprofloxacin (87.0%), Ampicillin/sulbactam (82.6%), Cefotaxime (81.5%), Ceftazidime (80.4%). Moreover, 50.0% of these isolates were non-susceptible to tigecycline. When evaluated by RAPD analysis, generated distinct band ranging in size from 1kb to 8k band varying from 4 to 10 bands. Stricter surveillance and more rapid detection are essential to prevent the spread of multi drug resistant A. baumannii.