• 제목/요약/키워드: agar dilution method

검색결과 118건 처리시간 0.025초

3 세대 세파계 항생제에 내성인 임상균주의 분포와 PCR 법을 이용한 TEM type $\beta$-lactamase 생산균주의 동정 (Prevalence of Strains Resistant to the Third Generation Cephalosporins among Clinical Isolates and Identification of TEM Type $\beta$-lactamase from Resistant Strains by PCR Method)

  • 김무용;오정인;송혜경;백경숙;곽진환
    • 약학회지
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    • 제39권3호
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    • pp.276-282
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    • 1995
  • Compared to the first and second-generation cephalosporins, the third-generation cephalosporins are remarkably stable against hydrolysis by the $\beta$-lactamases produced by aerobic gram-negative bacilli, such as Enterobacteriaceae. Among these bacteria, the most prevalent plasmid-encoded $\beta$-lactamase is TEM-1 $\beta$-lactamase belonging to class A or group 2b. This enzyme is produced constitutively and is principally active against peniciflins and old cephalosporins rather than third-generafion cephalosporins, carbapenems and mmobactams. However, new TEM type $\beta$-lactamases including TEM-9 and TEM-12 evolved through point mutations in a gene encoding $\beta$-lactamase have been discovered from patients during chemotherapy. These $\beta$-lactamases are known to be capable of hydrolyzing most of the third-generatim cephalosporins. To study the prevalence of $\beta$-lactamases from clinical isolates collected in Korea. the minimal inhibitory concentratims(MICs) of several third-generation cephalosporins against 628 clinical isolates were determined by agar dilution methods, and $\beta$-lactamas-producing bacteria were isolated by use of cefinase disc. By polymerase chain reaction (PCR) method, clinical isolates harboring a gene for TEM type $\beta$-lactamase were identified among the $\beta$-lactamase producing strains. Twentiy three percent of the clinical isolates was resistant to the thirdgeneration cephalosporins, and more than 90% of resistant cells produced various $\beta$-lactamases. TFM type $\beta$-lactamases were dominant in gram-negative bacilli, such as Escherichia coli, Klebsiella pneumoniae, Enterobacter species. These results suggest the necessity of the development of new cephalosporins which are stable against $\beta$-lactamases like TEM.

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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
    • 대한의생명과학회지
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    • 제26권2호
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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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    • 제44권1호
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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
    • 한국환경농학회지
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    • 제33권3호
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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.

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

  • 김윤태;김태운
    • 한국미생물·생명공학회지
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    • 제34권4호
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    • pp.342-351
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    • 2006
  • 병원내 항생제 다제 내성을 일으키는 CTX-M형 ESBL을 생성하는 E. coli와 Klebsielia pneumoniae의 생성현황을 조사하고 이들 균주로 인한 감염증치료와 역학적 조사연구에 도움이 되고자 효소의 유전형을 규명하였다. 2005년 7월-12월에 부산에 소재하고 있는 2개의 종합병원에서 분리된 E. coli와 K. pneumoniae 각각 153주, 52주를 수집하였다. 그 중에서 ESBL을 생성 하는 균주를 검출하기 위해 Double disk synergy test를 시행하여서 E. coli 23주와 K. pneumoniae 13주를 분리하였다. 균주의 동정은 Vitek system GNI card(bioMerieux Vitek Inc., Hazelwood, Mo., U.S.A.)로 확인하였고, 항생제감수성시험은 disk diffusion method 와 agar dilution method를 사용하였다. 분리된 균주들의 내성을 일으키는 ESBL유전형을 규명하기 위하여 Isoelectric focusing(IEF), polymerase chain reaction test, DNA sequencing을 시행하였다. A병원의 13주와 B병원의 10주로 총 23주의 E. coli(15.0%)와 A병원의 7주와 B병원의 6주로 K. pneumoniae 13주(25.0%)가 double disk synergy test 양성으로 ESBL 생성균주로 판정하였다. ESBL 생성 36균주를 대상으로 bla$_{TEM}$, bla$_{SHV}$, bla$_{CTX-M}$ 유전자 검출을 위한 PCR을 시행한 결과 bla$_{TEM}$ 유전자는 13주(36.1%), bla$_{SHV}$ 유전자는 13주(36.1%), bla$_{CTX-M}$ 유전자는 32주(88.9%)가 양성반응을 보여서 bla$_{CTX-M}$ 유전자를 가진 균주가 가장 많이 나타났다. 그리고, bla$_{TME}$, bla$_{SHV}$ 두 가지 유전자를 가지고 있는 균주는 1주(2.8%)만 나타났고 bla$_{TEM}$, bla$_{CTX-M}$두 가지 유전자를 가지고 있는 균주는 9주(25.0%), bla$_{SHV}$, bla$_{CTX-M}$ 두 가지 유전자를 가지고 있는 균주가 10주(27.8%)로 나타나 bla$_{CTX-M}$을 포함하는 복합유전자가 많이 증가함을 알 수 있었다. 또한 CTX-M형 ESBL을 생성하는 E. coli와 K. pneumoniae에 대한 cefutaxime의 MIC는 256 $\mu$g/m1 이상으로 ceftazidime의 16-256 $\mu$g/mL 이상보다 높은 분포를 보였다. 즉, CTX-M형 ESBL 유전자를 지닌 균주에 대한 cefotaxim의 MIC는 ceftazidime의 MIC에 비해서 상대적으로 높은 양상을 보였다. 이러한 결과는 국내의 대학병원 뿐 만 아니라 일반종합병원에서도 CTX-M형 ESBL 생성 E. coli와 K. pneumoniae가 존재하며 확산 중임을 시사한다. 앞으로 CTX-M형 ESBL의 만연과 변종 CTX-M형 ESBL의 출연을 감시하기 위한 정기적인 연구와 조사가 필요한 것으로 생각한다.

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

  • 김지연;이연태
    • 대한미생물학회지
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    • 제19권1호
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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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    • 제38권1호
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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.

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

  • 성기창;최원필
    • 대한수의학회지
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    • 제27권2호
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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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    • 제27권4호
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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.

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

  • 염수진;김승민;유연철;정희곤
    • 한국식품과학회지
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    • 제49권2호
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    • pp.146-150
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    • 2017
  • 본 연구에서는 지금까지 거의 보고된 바 없는 corosolic acid의 항균활성에 대해 대표적 식중독균 중 하나인 S. aureus를 대상으로 연구하였다. 대수기의 S. aureus에 corosolic acid를 처리하여 생장 저해 여부를 확인하였고, broth micro-dilution 방법과 agar disc diffusion 방법을 통해 corosolic acid의 S. aureus에 대한 항균활성을 측정해본 결과, corosolic acid의 농도에 비례하여 S. aureus 생장이 저해되는 것으로 확인되었다. 또한 corosolic acid의 존재 조건에서 S. aureus의 바이오필름 형성능이 MIC 보다 높은 농도뿐 아니라 낮은 농도에서 역시 저해되는 것으로 관찰되었다. 따라서 천연물질인 corosolic acid는 S. aureus에 의한 식품 오염을 억제하고 식중독 예방 및 잠재적 치료제로서 개발 될 가능성이 있는 것으로 판단되며, 나아가 다른 종류의 식중독균들에 대한 적용 가능 연구가 필요할 것으로 사료된다.