• Title/Summary/Keyword: Agar plate

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Detection of genetic mutations associated with macrolide resistance of Mycoplasma pneumoniae (Mycoplasma pneumoniae의 macrolide 내성과 연관된 유전자 변이의 검출)

  • Oh, Chi Eun;Choi, Eun Hwa;Lee, Hoan Jong
    • Clinical and Experimental Pediatrics
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    • v.53 no.2
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    • pp.178-183
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    • 2010
  • Purpose : The aim of this study was to identify mutations associated with macrolide resistance in Mycoplasma pneumoniae (MP) and to establish a cultural method to determine antimicrobial susceptibility. Methods : Nasopharyngeal aspirates (NPAs) were collected from 62 children diagnosed with MP pneumonia by a serologic method or polymerase chain reaction. The 23S rRNA and L4 ribosomal protein genes of MP were amplified and sequenced. To identify mutations in these 2 genes, their nucleotide sequences were compared to those of the reference strain M129. MP cultivation was carried out for 32 (28 frozen and 5 refrigerated) NPAs and M129 strain using Chanock's glucose broth and agar plate in a 5% $CO_2$ incubator at $37^{\circ}C$ and examined at 2-3 day intervals for 6 weeks. Results : Among the 62 specimens, 17 had M144V mutations in ribosomal protein L4. The A2064G mutation was observed in 1 specimen; its 23S rRNA gene was successfully sequenced. Culture for MP was successful from the M129 strain and 2 of the 5 NPAs that were refrigerated for no longer than 3 days. However, MP did not grow from the 28 NPAs that were kept frozen at $-80^{\circ}C$ since 2003. Conclusion : We found the M144V mutation of L4 protein to be common and that of domain V of 23S rRNA gene was relatively rare among MP. Studies on the prevalence of macrolide-resistant MP and the relationship between the mutations of 23S rRNA gene and ribosomal protein L4 will aid in understanding the mechanism of macrolide resistance in MP.

Pathogenicity and PCR detection of Vibrio tapetis in Manila clams, Ruditapes philippinarum (양식 바지락, Ruditapes philippinarum에 대한 Vibrio tapetis의 병원성과 PCR법에 의한 진단)

  • Park, Sung-Woo;Lee, Kyung-Hee
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.39-48
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    • 2005
  • Pathogenicity of Vibrio tapetis, the causative bacterium of 'brown ring disease (BRD)' was evaluated in Manila clams (Ruditapes philippinarumi by artificially 0.1 $m\ell$ infection of $1.0\times10^5$cells and $1.0\times10^8$ cells at 20 $^{\circ}C$. A PCR assay based on 16S rRNA to detect the bacteria in clam tissues was established. Accumulative mortality of clams infected with $1.0\times10^7$cells and $1.0\times10^4$ cells per an individual of the bacteria was 67.5% and 7.5%, respectively. However, the deposit of brown pigment in the inner shells by accumulation of chonchiolin was not found. The bacteria were not be able to re-isolate from the infected clams by the conventional agar plate method but were easily detected by PCR assay established in this experiment. In clams artificially infected with 10 species of Vibrio, a 414bp for V. tapetis was detected in PCR assay. The specific band in the clams infected with $1.0\times10^4$cells per an individual of V. tapetis was detected only in gills one day after the infection but never be found in any tissues including gills three days after the infection. In the case of clams infected with $1.0\times10^8$cells per an individual of V. tapetis the specific band was detected in gills and intestine one day after the infection, in all tissues three days after the infection, and then in gills and adductor muscle nine days after the infection. The PCR assay was applied to detect V. tapetis in manila clam, surf clam (Mactra veneriformis), oyster (Crassostrea gigas) and Thomas' rapa whelk (Rapana venosa) taken from Taean and Gochang from April to July 2004. The infection rates were detected to 23.1% and 9.4% in the oyster and surf clam, while manila clam and Thomas' rapa whelk were not found.

DETECTION OF METHICILLIN OR VANCOMYCIN-RESISTANT STAPHYLOCOCCUS AUREUS FROM DENTAL HOSPITAL (치과병원 진료실 내에서 메티실린 또는 반코마이신 저항성 Staphylococcus aureus의 검출)

  • Min, Jung-Hee;Park, Soon-Nang;Hwang, Ho-Keel;Min, Jung-Beum;Kim, Hwa-Sook;Kook, Joong-ki
    • Restorative Dentistry and Endodontics
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    • v.32 no.2
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    • pp.102-110
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    • 2007
  • The purpose of this study was to obtain the basic information for the improvement of dental environment by investigating the presence of methicillin- or vancomycin-resistant Staphylococcus aureus (MRSA or VRSA) isolated from dental health care workers (DHCWs) and environment of the Chosun University Dental Hospital (CUDH) and a private dental clinic (control group). Staphylococcus aureus (S. aureus) was isolated from anterior nares of 42 DHCWS and 38 sites, unit chairss, x-ray devices, computers, etc., at 10 departments of the CUDH and 20 DHCWs and 11 sites at the private dental clinic. S. aureus was isolated on mannitol salt agar plate and confirmed by PCR with S. aureus species-specific primer. Antimicrobial susceptibility test of clinical isolates of S. aureus against several antibiotics including methicillin (oxacillin) was performed by investigating minimum inhibitory concentration (MIC) using broth microdilution assay. In addition, PCR was performed to detect the methicillin- or vancomycin-resistant gene. The data showed that one strain of S. aureus was isolated from DHCWs of the CUDH and three strains of S. aureus was isolated from 3 samples of the private dental clinic, respectively. All of the isolates from the CUDH and the private dental clinic had resistance to penicillin G, amoxicillin and vancomycin and susceptibility to oxacillin and ciprofloxacin. The S. aureus strains were already obtained the resistance to penicillin G and amoxicillin. These results suggest that two dental clinics were under relatively safe environment.

Bactericidal Efficacy of Fumagari OPP®, Fumigant Against Staphylococcus aureus (훈증소독제, Fumagari OPP®의 Staphylococcus aureus에 대한 살균효과)

  • Cha, Chun-Nam;Park, Eun-Kee;Choi, Hyunju;Kim, Yongpal;Yoo, Chang-Yeol;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.28 no.4
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    • pp.349-353
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    • 2013
  • This study was performed to evaluate the bactericidal efficacy of Fumagari OPP$^{(R)}$, fumigation disinfectant, containing 20% ortho-phenylphenol against Staphylococcus aureus (S. aureus). In this research, efficacy test of fumigant against S. aureus was carried out according to French standard NF T 72-281. S. aureus working culture suspension number (N value), all of the colony numbers on the carriers exposed with the fumigant (n1, n2, and n3), the number of bacterial test suspentions by pour plate method (N1), the number of bacterial test suspentions by filter membrane method (N2) and the mean number of bacteria recovered on the control-carriers (T value) were obtained from the preliminary test. In addition, the reduction number of S. aureus exposed with the fumigant (d value) was calculated using T value, the mean number of bacteria in recovery solution (n'1) and the mean number of bacteria on carriers plated in agar (n'2). N value was $4.0{\times}10^8$ CFU/mL, and n1, n2, and n3 were higher than 0.5N1, 0.5N2 and 0.5N1, respectively. Additionally, T value was $3.4{\times}10^6$ CFU/mL. In the bactericidal effect of the fumigant, the d value was 6.43 logCFU/mL. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. The results indicated that Fumagari OPP$^{(R)}$ had an effective bactericidal activity against S. aureus, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with pathogenic bacteria.

Surface roughness and $Candida$ $albicans$ adhesion to flexible denture base according to various polishing methods (연마방법에 따른 탄성의치의 표면거칠기와 $Candida$ $albicans$의 부착율 변화)

  • Oh, Ju-Won;Seo, Jae-Min;Ahn, Seung-Keun;Park, Ju-Mi;Kang, Cheol-Kyun;Song, Kwang-Yeob
    • The Journal of Korean Academy of Prosthodontics
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    • v.50 no.2
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    • pp.106-111
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    • 2012
  • Purpose: The purpose of this study was to compare the effect of 3 chairside polishing methods and laboratory polishing methods on surface roughness and $C.$ $albicans$ adhesion of polyamide denture base. Materials and methods: Using contact profilometer, the surface of polyamide specimens ($25{\times}15{\times}2mm$) was studied after conventional polishing without finishing and after chiarside polishing with 2 chiarside polishing kits and chairside-pumice polishing following finishing with tungsten carbide bur. To evaluate the adhesion of $C.$ $albicans$, $C.$ $albicans$ suspension was overlayed on the test specimen. And the specimens were incubated for 2 hours. Imprint culture method was achieved and counted the colony on the agar plate. Polished polyamide were evaluated using a scanning electron microscope. The statistics were conducted using one-way ANOVA and in case of difference, Scheffe test and Tamhane's T2 test were used. Results: Surface roughness (Ra) of surfaces polished with 2 chairside polishing kits had higher than conventional polishing and pumice polishing. The highest roughness value was $0.32{\pm}0.10{\mu}m$, and the lowest was $0.02{\pm}0.00{\mu}m$. The adhesion of $C.$ $albicans$ on the specimens polished with chairside polishing group and pumice polishing group were increased than conventional polishing group ($P$<.01). Conclusion: Conventional laboratory polishing was found to produce the smoothest surface and the lowest adhesion of $C.$ $albicans$. Two groups polished with Chairside polishing kits were similar with respect to surface roughness. Surface of the specimen polished with pumice is significantly smoother than 2 chairside polishing groups, but the result of $C.$ $albicans$ adhesion is that group polished with pumice was similar with 2 chairside polishing groups ($P$>.01).

Expression and Purification of Three Lipases (LipAD1, LipAD2, and LipAD3) and a Lipase Chaperone (LipBD) from Acinetobacter schindleri DYL129 (Acinetobacter schindleri DYL129 유래의 3개 lipases와 chaperone의 발현과 정제)

  • Kim, Sun-Hee;Lee, Yong-Suk;Jeong, Hae-Rin;Pyeon, Hyo-Min;You, Ju-Soon;Choi, Yong-Lark
    • Journal of Life Science
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    • v.29 no.4
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    • pp.492-498
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    • 2019
  • Previously, three kinds of lipases, lipAD1, lipAD2, and lipAD3, and lipase chaperone, lipBD, of Acinetobacter schindleri DYL129 isolated from soil sample were reported. In this report, three lipase and lipase chaperone were cloned into the pET32a(+) or pGEX-6P-1 vectors for protein expression in Escherichia coli, and named as pETLAD1, pETLAD2, pETLAD3 and pETLBD or pGEXLAD1, pGEXLAD 2, pGEXLAD3 and pGEXLBD, respectively. Protein expression rate was higher in pET system than in pGEX system. Although LipAD1 and LipAD2 were produced as inclusion bodies, their expression levels were high. So LipAD1 and LipAD2 could be solubilized in 8 M urea buffer and purified. LipAD3 and LipBD were overexpressed in soluble form and purified. Those proteins were purified by His-tag affinity chromatography connected in AKTA prime system. The activities of the purified lipases were demonstrated with 1% tributyrin agar plate. After purification, molecular mass was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. LipAD1 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl butyrate, LipAD2 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl myristate, and LipAD3 showed high activity toward ${\rho}$-nitrophenyl acetate, ${\rho}$-nitrophenyl butyrate, and ${\rho}$-nitrophenyl miristate, respectively. Three lipases, LipAD1, LipAD2, and LipAD3, showed optimal reaction at $50^{\circ}C$ using ${\rho}$-nitrophenyl butyrate, as substrate.

Studies on Biological Activity of Woad Extractives (XV) - Antimicrobial and antioxidative activities of extracts from diverse families - (수목 추출물의 생리활성에 관한 연구(XV) - 과별(科別)에 따른 항균 및 항산화 활성 -)

  • Lee, Sung-Suk;Lee, Hak-Ju;Choi, Don-Ha
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.4
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    • pp.8-17
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    • 2004
  • Antifungal, antibacterial, and antioxidative activities of ethanol extracts from 65 families 263 species were investigated to select tree species for the utilization of natural fungicide or preservative resources. The antifungal activities of extracts from wood, leaf and bark were measured as hyphal growth inhibition rate using four plant pathogenic and five wood rotting fungi. High inhibitory effect on the fungi growth was found in five species of Pinaceae (Pinus koraiensis, P. rigida, P. densiflora, P. banksiana. Cedrus deodara), three species of Cupressaceae (Juniperus rigida, J. chinensis, Chamaecyparis obtusa) and three species of Leguminosae (Albizzia julibrisssin, Sophora japonica, Maackia amurensis), respectively. Antibacterial activities of ethanol extracts were determined by means of disc-agar plate diffusion method using three gram-positive and five gram-negative bacteria. The ethanol extracts, which showed prominent effect on the suppression of bacteria growth, were six species of Betulaceae (Carpinus tschonoskii, C. coreana, C. laxiflora, Alnus hirsuta, A. firma, Betula schmidtii), five species of Fagaceae (Castanopsis cuspidata var. sieboldii, Quercus serrata, Q. mongolica, Q aliena, C crenata), four species of Euphorbiaceae (Aleurites fordii, Sapium sebiferum, S japonicum Mallotus japonicus) and three species of Elaeagnaceae (Elaeagnus umbellata, Elaeagnus glanbra, Elaeagnus macrophylla). According to these results, the extracts from Zelkova serrata, Pinus densiflora, Maackia amurensis, Chamaecyparis obtusa and Juniperus chinensis could be available for natural fungicide or food preservatives, because ethanol extracts from these species indicated excellent antifungal and antibacterial activities. In order to test antioxidative activities of ethanol extracts, free radical scavenging method was adopted with 1,1-diphenyl-2-picrylhydrohydrazyl (DPPH). Free radical scavenging activity was proved very high in the extracts of eight species of Rosaceae (Eriobotrya japonica, Prunus takesimensis, P yedoensis, P padus, P armeniaca var. ansu, Chaenomeles sinensis, Stephanandra incisa, Rosa multiflora) and five species of Ericaceae (Rhododenron mucronulatum, R. scblippenbacbii, R. yedoense var. poukhanense, Vaccinium bracteatum, V oldbami), resvectively. It turned out from this study that only six species among 48 species of Rosaceae showed less than 80% free radical scavenging activity. As a consequences, it could be deduced that the components effective on antioxidative activity commonly exist in Rosaceae plant family.