• Title/Summary/Keyword: Agar dilution method

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Comparison of Fluoroquinolone Resistance Determinants in Uropathogenic Escherichia coli between 2 Time Periods of 1989 and 2010-2014 at Gangwon Province in Korea

  • Park, Min
    • Biomedical Science Letters
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    • v.26 no.2
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    • pp.120-126
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    • 2020
  • Fluoroquinolone (FQ) resistant uropathogenic Escherichia coli (UPEC) have become a major problem in urinary tract infections (UTIs). The purpose of this study was to compare the quinolone resistance-determining region (QRDR) and plasmid mediated quinolone resistance (PMQR) determinants of FQ resistant UPEC between 1989 and 2010-2014. A total of 681 strains of UPEC clinical isolates was collected from Korean healthcare facility in 1989 (123 strains) and in 2010-2014 (558 strains). The minimum inhibitory concentrations (MICs) of FQs were determined by agar dilution method. QRDRs (gyrA, gyrB, parC and parE) and PMQR determinants (qnrA, qnrB, qnrS, aac(6')-Ib-cr and qepA) were analyzed polymerase chain reaction and sequencing method. Among 681 isolates, FQ resistant UPEC were 3 strains (2.4%) in 1989 isolates and 220 strains (39.4%) in 2010-2014 isolates. The rate of the FQ resistant UPEC strains in 2010-2014 isolates was increased than that of in 1989 isolates. UPEC isolates from 1989 and 2010-2014 were shown to carry mutations in gyrA (Ser83 and Asp87), gyrB (Ser464 and Thr469), parC (Ser80 and Glu84) and parE (Glu460, Ser458, Ile464 and Leu445). The most common mutations of QRDRs in 1989 isolates were Ser83Leu and Asp87Gly in gyrA and Ser80Ile in parC (2 strains: 66.7%) while those in 2010-2014 isolates were Ser83Leu and Asp87Asn in gyrA and Ser80Il2 and Glu84Val in parC (88 strains: 40.0%). PMQR determinants were detected only in 2010-2014 UPEC strains (47 strains: 21.4%).

In vitro evaluation of octenidine as an antimicrobial agent against Staphylococcus epidermidis in disinfecting the root canal system

  • Chum, Jia Da;Lim, Darryl Jun Zhi;Sheriff, Sultan Omer;Pulikkotil, Shaju Jacob;Suresh, Anand;Davamani, Fabian
    • Restorative Dentistry and Endodontics
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    • v.44 no.1
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    • pp.8.1-8.7
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    • 2019
  • Objectives: Irrigants are imperative in endodontic therapy for the elimination of pathogens from the infected root canal. The present study compared the antimicrobial efficacy of octenidine dihydrochloride (OCT) with chlorhexidine (CHX) and sodium hypochlorite (NaOCl) against Staphylococcus epidermidis (S. epidermidis) for root canal disinfection. Materials and Methods: The minimum inhibitory concentration (MIC) was obtained using serial dilution method. The agar diffusion method was then used to determine the zones of inhibition for each irrigant. Lastly, forty 6-mm dentin blocks were prepared from human mandibular premolars and inoculated with S. epidermidis. Samples were randomly divided into 4 groups of 10 blocks and irrigated for 3 minutes with saline (control), 2% CHX, 3% NaOCl, or 0.1% OCT. Dentin samples were then collected immediately for microbial analysis, including an analysis of colony-forming units (CFUs). Results: The MICs of each tested irrigant were 0.05% for CHX, 0.25% for NaOCl, and 0.0125% for OCT. All tested irrigants showed concentration-dependent increase in zones of inhibition, and 3% NaOCl showed the largest zone of inhibition amongst all tested irrigants (p < 0.05). There were no significant differences among the CFU measurements of 2% CHX, 3% NaOCl, and 0.1% OCT showing complete elimination of S. epidermidis in all samples. Conclusions: This study showed that OCT was comparable to or even more effective than CHX and NaOCl, demonstrating antimicrobial activity at low concentrations against S. epidermidis.

Phytoextraction of Heavy Metals Induced by Bioaugmentation of a Phosphate Solubilizing Bacterium

  • Arunakumara, K.K.I.U.;Walpola, Buddhi Charana;Song, Jun-Seob;Shin, Min-Jung;Lee, Chan-Jung;Yoon, Min-Ho
    • Korean Journal of Environmental Agriculture
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    • v.33 no.3
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    • pp.220-230
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    • 2014
  • BACKGROUND: Excessive metals in the soil have become one of the most significant environmental problems. Phytoremediation has received considerable attention as a method for restoring the contaminated soils. The microbes having remarkable metal tolerance and plant growth-promoting abilities could also play a significant role in remediation of metal-contaminated soils, because bioaugmentation with such microbes could promote phytoextraction of metals. Therefore, the present study was focused on evaluating the phytoextraction of heavy metals (Co, Pb and Zn) in Helianthus annuus (sunflower) induced by bioaugmentation of a phosphate solubilizing bacterium. METHODS AND RESULTS: A phosphate solubilizing bacterium was isolated from metal-contaminated soils based on the greater halo size (>3 mm) with solid NBRIP agar medium containing 10 g glucose, 5 g $Ca_3(PO_4)_2$, 5 g $MgCl_2{\cdot}6H_2O$, 0.25 g $MgSO_4.7H_2O$, 0.2 g KCl, 0.1 g $(NH_4)_2SO_4$ in 1 L distilled water. Isolated bacterial strain was assessed for their resistance to heavy metals; $CoCl_2.6H_2O$, $2PbCO_3.Pb(OH)_2$, and $ZnCl_2$ at various concentrations ranging from $100-400{\mu}g/mL$ (Co, Pb and Zn) using the agar dilution method. A pot experiment was conducted with aqueous solutions of different heavy metals (Co, Pb and Zn) to assess the effect of bacterial strain on growth and metal uptake by Helianthus annuus (sunflower). The impact of bacterial inoculation on the mobility of metals in soil was investigated under laboratory conditions with 50 mL scaled polypropylene centrifuge tubes. The metal contents in the filtrate of plant extracts were determined using an atomic absorption spectrophotometer (Perkinelmer, Aanalyst 800, USA). CONCLUSION: Inoculation with Enterobacter ludwigii PSB 28 resulted in increased shoot and root biomass and enhanced accumulation of Co, Pb and Zn in Helianthus annuus plants. The strain was found to be capable of promoting metal translocation from the roots to the shoots of H. annuus. Therefore, Enterobacter ludwigii PSB 28 could be identified as an effective promoter of phytoextraction of Co, Pb and Zn from metal-contaminated soils.

Prevalence of CTX-M-type Extended-Spectrum $\beta$-Lactamases Producing Escherichia coli and Klebsieilla pneumoniae Isolates in General Hospitals in 2005 (임상에서 분리된 CTX-M형 Extended-Spectrum $\beta$-Lactamases를 생산하는 Escherichia coli와 Klebsiella pneumoniae의 유행)

  • Kim, Yun-Tae;Kim, Tae-Un
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.342-351
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    • 2006
  • The aim of this study was to survey susceptibilities of Escherichia coli and Klebsiella pneumoniae isolates against cefotaxime and to determine the prevalences of CTX-M type extended-spectrum $\beta$-lactamases (ESBLs) producing E. coli and K. pneumoniae in Korea. During the period of February to July, 2005, 153 E. coli and 52 K. neumoniae isolates were collected from 2 hospitals in Busan. Antimicrobial susceptibilities to cefotaxime were tested by the disk diffusion method. ESBL production of E. coli and K. pneumoniae was determined by the double disk synergy test. MICs of $\beta$-lactam antibiotics were determined by the agar dilution method. Blac$_{CTX-M}$ genes of the organism were detected by PCR. Among 153 isolates of E. coli and 52 isolates of K. neumoniae, 27 (17.6%) and 25 (48.0%) were intermediate or resistant to cefotaxime, respectively. Twenty-three (15.0%) isolates out of 153 E. coli and 13 (25.0%) out of 52 K. neumoniae isolates showed positive results for ESBL by the double disk synergy test. Twenty isolates out of 23 ESBL producing E. coli and 12 out of 13 ESBL producing K. neumoniae isolates harbored biacTx-M gene,11 of ESBL producing E. coli and 12 of ESBL producing K. neuinoniae isolates harbored bla$_{CTX-M}$ gene, 11 of the ESBL producing E. coli and 2 of ESBL producing K. neumoniae isolates harbored bla$_{TEM}$ gene, and 1 of the ESBL producing E. coli and 12 of ESBL producing K. neumoniae isolates harbored bla$_{SHV}$ gene. E. coli and K. neumoniae isolates producing CTX-M-type ESBLs were not uncommon in Korea. It is thought that continuous survey are necessary for inspecting the spread and novel variants of CTX-M-type ESBL genes. Further me]'e investigation and research on ESBL producing strains are needed in order to prevent the spread of resistant bacteria.

Drug Resistance and R-Plasmids of Shigella Strains Isolated from Humans, Korea (Shigella균속의 항균제내성 및 전달성 R-Plasmid에 관한 연구)

  • Kim, Ji-Youn;Lee, Yun-Tai
    • The Journal of the Korean Society for Microbiology
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    • v.19 no.1
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    • pp.11-24
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    • 1984
  • Shigella remains to be an important enteric pathogen in this country for the present. Moreover, most of the isolates have become multiple resistant to various antibiotics which used to be drugs of choice for shigellosis. This study was made as an attempt to assess the present stage of antibiotic resistance and the incidence and transferability of R factors of Shigella. A total of one hundred and seventeen strains of Shigella isolated from patients in Seoul and provincial area between 1982 and 1983 were tested for their resistant to antimicrobial agents and transmission of R-plasmid. Antibiotic susceptibilities were determined by an agar dilution method. Muller hinton agar were used for the assay of drug resistance and tryptic soy broth were used for propagating medium for conjugation. Shigella isolated found to be one or more antibiotics were considered potential donor of R-plasmid. The following results were obtained. 1. Among 117 strains of Shigella isolated, 111 strains(94.9%) were found to be resistant to one or more drugs tested and 97.3% of these resistant strains were multiply resistant, indicating the multiply resistant strains were more than the single resistant strains. Only six strains were susceptible to all drugs tested. 2. Among 117 strains of Shigella isolated, 107 strains(91.5%) were resistant to Tetracyclin(Tc), 106 strains(90.6%) to Chloramphenicol(Cp) and Streptomycin(Sm), 97 strains(82.9%) to Ampicillin(Ap), 68 strains(58.1%) to Cephaloridine(Cr), 10 strains(8.5%) to Nalidixic acid(Na), 5 strains(4.3%) to Kanamycin(Km) and 2 strains(1.7%) to Rifampicin. No strain was resisfant to Amikacin(Ak) and Gentamicin(Gm). 3. All drug-resistant Shigella strains, except three, were multiply resistant to two or more drugs. Fifty eight strains were resistant to five drugs, followed by 26 strains resistant to dour drugs, 12 strains resistant to three drugs and 11 strains resistant to six drugs. 4. The 73% of multiply drug-resistant Shigella transferred their resistance to E. coli by conjugation and the resistance was considered to be mediated by R-plasmid. Resistance to Nalidixic acid and Rifampicin were not transferred by conjugation to recipient. As for the transferability of resistance to each seperate drug, Ap resistance was transferred with 73.2% frequence and Cm and Tc resistance were transferred with approximately 50-60% frequence whereas Sm and Cr resistance were transferred in 19.1-21.4% The other four drugs resistant failed to transfer their resistance to recipient. 5. As for the incidence and transferability of resistance to each seperate drug, the strains resistant to Tc and Cm were encountered most frequently with the rate of 91-92%, whereas transfer of Tc and Cm were low, 51-52%. The incidence of Sm resistance was very high(90.6%) but transferability of drugs resistance was much lower(25.4%). Though the incidence of Km reristance was much lower(4.3%) transferability of Km resistance was considerably higher(60%). 6. The greater the multiplicity of resistance, the greater was the likelihood that part of all of the resistance markers would be transferable.

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Inhibitory effect on Streptococcus mutans and mechanical properties of the chitosan containing composite resin

  • Kim, Ji-Sun;Shin, Dong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.38 no.1
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    • pp.36-42
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    • 2013
  • Objectives: This study evaluated the antibacterial effect and mechanical properties of composite resins ($L_{CR}$, $M_{CR}$, $H_{CR}$) incorporating chitosan with three different molecular weights (L, Low; M, Medium; H, High). Materials and Methods: Streptococcus (S). mutans 100 mL and each chitosan powder were inoculated in sterilized 10 mL Brain-Heart Infusion (BHI) solution, and was centrifuged for 12 hr. Absorbance of the supernatent was measured at $OD_{660}$ to estimate the antibacterial activities of chitosan. After S. mutans was inoculated in the disc shaped chitosan-containing composite resins, the disc was cleansed with BHI and diluted with serial dilution method. S. mutans was spread on Mitis-salivarius bacitracin agar. After then, colony forming unit (CFU) was measured to verify the inhibitory effect on S. mutans biofilm. To ascertain the effect on the mechanical properties of composite resin, 3-point bending and Vickers hardness tests were done after 1 and 3 wk water storage, respectively. Using 2-way analysis of variance (ANOVA) and Scheffe test, statistical analysis was done with 95% significance level. Results: All chitosan powder showed inhibition effect against S. mutans. CFU number in chitosan-containing composite resins was smaller than that of control resin without chitosan. The chitosan containing composite resins did not show any significant difference in flexural strength and Vickers hardness in comparison with the control resin. However, the composite resin, $M_{CR}$ showed a slightly decreased flexural strength and the maximum load than those of control and the other composite resins $H_{CR}$ and $L_{CR}$. Conclusions: $L_{CR}$ and $H_{CR}$ would be recommended as a feasible antibacterial restorative due to its antibacterial nature and mechanical properties.

Characterization of Yersinia Species Isolated from Animals in Korea (동물(動物)에 있어서 Yersinia속균(屬菌)의 분포(分布)와 특성(特性)에 관(關)한 연구(硏究))

  • Sung, Ki-chang;Choi, Won-pil
    • Korean Journal of Veterinary Research
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    • v.27 no.2
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    • pp.235-243
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    • 1987
  • This paper deals with the distribution of Yersinia spp. isolated from the feces or the cecal contents of 1,755 pigs, 558 cows, 428 pigs slaughtered, 271 dogs slaughtered and 91 deer during the period of March 1985 to February 1986. Isolated Yersinia spp. were examined for serotype, biotype and antibiotic susceptibility of Y. enterocolitica. The results were as follows; One hundred and fourty-three stains of Yersinia spp. were isolated from 141(4.5%) out of 3,103 animals examined and their isolates were identified as Y. enterocolitica(138 strains), Y. kristensenii (3 strains), Y. intermedia(1 strain) and Y. pseudotuberculosis(1 strain). Yersinia spp. were isolated from 122(7.0%) of 1,755 pigs in piggeries, 15(3.5%) of 428 pigs slaughtered and 4(1.5%) of 271 dogs slaughtered, but no Yersinia spp. were isolated from cows and deer. The isolation rate of Yersinia spp. in pigs ranged from 5.9~8.0% in piggeries, it was higher in summer and autumn and highest in fattening pigs groups(10.4%), especially. One hundred and thirty-eight Y. enterocolitica isolates belonged to serotype 0 : 3(95 strains), 0 : 8(13 strains), 0 : 5(7 strains), 0 : 9(6 strains), 0 : 1, 2(1 strain) and untypable(16 strains), among them strains of serotype 0 : 3 biotype 3B(91 strains) were predominant. Antibiotic susceptibility test of 138 isolates of Yersinia spp. was performed by the agar dilution method, using 8 antibiotics as follows: ampicillin(Am), chloramphenicol, kanamycin, nalidixic acid(Na), rifampicin(Rf), streptomycin, sulfadimethoxine(Su) and tetracycline. All the strains tested were susceptible to Rf and Na, but resistant to Su, and 136 strains(98.6%) were also resistant to Am.

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Genomic Analysis of a Freshwater Actinobacterium, "Candidatus Limnosphaera aquatica" Strain IMCC26207, Isolated from Lake Soyang

  • Kim, Suhyun;Kang, Ilnam;Cho, Jang-Cheon
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.825-833
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    • 2017
  • Strain IMCC26207 was isolated from the surface layer of Lake Soyang in Korea by the dilutionto-extinction culturing method, using a liquid medium prepared with filtered and autoclaved lake water. The strain could neither be maintained in a synthetic medium other than natural freshwater medium nor grown on solid agar plates. Phylogenetic analysis of 16S rRNA gene sequences indicated that strain IMCC26207 formed a distinct lineage in the order Acidimicrobiales of the phylum Actinobacteria. The closest relative among the previously identified bacterial taxa was "Candidatus Microthrix parvicella" with 16S rRNA gene sequence similarity of 91.7%. Here, the draft genome sequence of strain IMCC26207, a freshwater actinobacterium, is reported with the description of the genome properties and annotation summary. The draft genome consisted of 10 contigs with a total size of 3,316,799 bp and an average G+C content of 57.3%. The IMCC26207 genome was predicted to contain 2,975 protein-coding genes and 51 non-coding RNA genes, including 45 tRNA genes. Approximately 76.8% of the protein coding genes could be assigned with a specific function. Annotation of the IMCC26207 genome showed several traits of adaptation to living in oligotrophic freshwater environments, such as phosphorus-limited condition. Comparative genomic analysis revealed that the genome of strain IMCC26207 was distinct from that of "Candidatus Microthrix" strains; therefore, we propose the name "Candidatus Limnosphaera aquatica" for this bacterium.

Inhibition of growth and biofilm formation of Staphylococcus aureus by corosolic acid (Corosolic acid에 의한 Staphylococcus aureus의 생장 및 생물막 형성 저해)

  • Yum, Su-Jin;Kim, Seung Min;Yu, Yeon-Cheol;Jeong, Hee Gon
    • Korean Journal of Food Science and Technology
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    • v.49 no.2
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    • pp.146-150
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    • 2017
  • Staphylococcus aureus is a pathogenic bacterium that causes food poisoning, exhibits a strong capacity to form biofilm, and is highly resistant to antimicrobial agents. The purpose of this study was to investigate the antimicrobial characteristics of corosolic acid against S. aureus. S. aureus showed high susceptibility to corosolic acid in a concentration-dependent manner. The minimum inhibitory concentration and colony-forming ability determined by the broth microdilution method showed that corosolic acid had strong antimicrobial activity against the bacteria. The diameters of the inhibition zone and numbers of colony forming units at each concentration of corosolic acid were also measured. In addition, corosolic acid displayed potent biofilm inhibition activity against S. aureus at concentrations below its minimum inhibitory concentration. These results suggest that corosolic acid can be used to effectively prevent biofilm formation by S. aureus, thereby making S. aureus more susceptible to the action of antimicrobials.

Ampicillin and co-trimoxazole susceptibility of Shigella isolated in 1978-80 (Shigella $1978{\sim}80$년 분리주의 Ampicillin 및 Co-trimoxazole 감수성)

  • Chong, Yun-Sop;Lee, Sam-Uel Y.;Ryu, Young-Hat
    • The Journal of the Korean Society for Microbiology
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    • v.15 no.1
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    • pp.33-38
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    • 1980
  • Shigella remains to be an important enteric pathogen in this country for the moment. Moreover, since 1978, most of the isolates have become resistant to ampicillin and co-trimoxazole, which used to be the drugs of choice for shigellosis. Since a disc diffusion technique alone has been used in our routine susceptibility test, the minimum inhibitory concentrations(MIC) of both ampicillin and co-trimoxazole to Shigella have never been known. In order to determine these, 195 isolates were tested by an agar dilution method, all of which were isolated at Yonsei Medical Center during the period of June 1978 to July 1980. The following results were obtained. 1. Sixty cultures(29.7%) were susceptible to ampicillin, being the MIC of 8 ${\mu}g/ml$ or less and 53(27.2%) were susceptible to co-trimoxazole, being the MIC of TMP/SMZ 4/76 ${\mu}g/ml$ or less. S. flexneri type 2 was often resistant to both antimicrobic agents. 2. An increasing rate of resistant isolates was noted, particularly in the year of 1979. 3. Many isolates were resistant to both agents. Somewhat more cultures. were ampicillin susceptible and co-trimoxazole resistant than the other way around. It seems that the determination of species or even serotypes might be of help sometimes to select proper antimicrobic agent to control the infection. A routine antimicrobic susceptibility test of Shigella to both ampicillin and co-trimoxazole would be advisable for a better selection of chemotherapeutic agent.

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