• Title/Summary/Keyword: enrichment culture

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Evaluation of the Selective Enrichment Culture to Recover Clostridium difficile

  • An, Byoungrak;Kim, Heejung;Lee, Kyungwon
    • Korean Journal of Clinical Laboratory Science
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    • v.46 no.4
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    • pp.140-142
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    • 2014
  • To evaluate the recovery rates to increase toxigenic C. difficile, the selective enrichment broth culture methods were compared with commonly used cytotoxin assays and toxigenic culture. First, the enrichment culture, using the selective medium broth for 2 to 5 days, was performed and then, toxigenic C. difficile was confirmed by C. difficile toxin gene-specific PCR after being cultured on C. difficile selective agar. The sensitivity of C. difficile from the enrichment culture (100%) was higher than that of C. difficile selective agar culture (93.8%), while positive predictive values (PPV) were low; 72.7% (16/22) and 88.2% (15/17). PPV of the enrichment culture are not high. Recently, combinations of C. difficile selective agar culture, C. difficile A & B assays, glutamate dehydrogenase, and nucleic acid amplification method are widely used. The enrichment culture was disadvantageous in PPV, turn-around time, and cost. So, what we performed is not considered as a common method of diagnosis of C. difficile-associated diarrhea.

혐기성 PCE 탈염소화 관련 미생물 군집 특성

  • 이태호;문부영;박태주
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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    • pp.133-137
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    • 2004
  • Tetrachloroethylene(PCE) dechlorination was investigated in an anaerobic enrichment culture from landfill soil. Anaerobic PCE dechlorinating microorganisms could convert 150mg/L of PCE via trichloroethylene(TCE) to cir-1,2-dichloroethylene(CDCE) within 2 days at the optimum temperature of 30 to 35$^{\circ}C$. The enrichment culture could dechlorinate TCE but did not degrade other chlorinated aliphatic compounds, such as cDCE, trans-1,2-dichloroethylene, 1,1-dichloroethylene, 1,1-dichloroethane, 1,2-dichloro- ethane, and 1,1,1-trichloroethane during 5 days incubation. Several isolates from the enrichment culture did not show dechlorinating activity of PCE. Microbial analysis of the dechlorinating enrichment culture by using Polymerase chain reaction-Denaturing gradient gel electrophoresis (PCR-DGGE) method showed that at least three microorganisms were related to the anaerobic PCE dechlorination in the enrichment

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Salmonella sp. 의 신속한 동정을 위한 증진배양의 개선에 관한 연구

  • Kim, Kee-Tae;Kim, Tae-Ue;Yook, Soon-Hak;Pek, Un-Hua
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.647-651
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    • 1996
  • The development of an enrichment method for the rapid and effective identification of Salmonella spp. in sewage or food was studied. As a growth factor for Salmonella, 10 mM cyclic adenosine monophosphate (cAMP) in trypticase soy broth with 0.6% yeast extract (TSBYE) increased cell number five-folds and 0.6% yeast extract in selenite broth increased cell number ten-folds of control. Bile salts in selenite broth was tested for the selection of S. enteritidis in a mixture with Staphylococcus aureus, Pseudomonas aeruginosa, Lactobacillus plantarum and Escherichia coli. The latter four strains were effectively inhibited at 0.1% bile salt. A two-step culture method was used to enrich Salmonella spp.; a primary-enrichment and secondary- enrichment culture. At a primary-enrichment step, selenite broth with 0.6% yeast extract and 10 mM cAMP was used, and at a secondary-enrichment step, 0.1% bile salt was additionally used. Culture times of a primary- enrichment and a secondary-enrichment step were 8 hr and 6 hr, respectively. In this procedure, cell number increased from 10$^{0.3}$ to 10$^{8.5}$ with inhibition of other strains within 14 hr. In the case of an initial cell concentrarion as low as 10$^{-2}$ cfu/ml, a cell number increased to 10$^{7}$ cfu/ml by using a 10 hr primary-enrichment and 6 hr secondary-enrichment procedure.

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Analysis of Archaeal Community in Autotrophic Perchlorate-degrading Enrichment Culture (독립영양 방식으로 퍼클로레이트를 분해하는 농화배양 내 고세균 군집 분석)

  • Kim, Young-Hwa;Do, Sanghyun;So, Hyunseung;Been, Junwon;Sung, Haechan;Ji, Sungchan;Son, Myunghwa;Ahn, Yeonghee
    • Journal of Life Science
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    • v.27 no.4
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    • pp.435-441
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    • 2017
  • Perchlorate ($ClO_4^-$) is an emerging contaminant detected in soil, groundwater, and surface water. Previous study revealed bacterial community in the enrichment culture tdegraded perchlorate using elemental sulfur as an electron donor. Quantitative and qualitative molecular methods were employed in this study to investigate archaeal community in the enrichment culture. Real-time qPCR showed that archaeal 16S rRNA gene copy number in the culture was about 1.5% of bacterial 16S rRNA gene copy number. This suggested that less archaea were adapted to the environment of the enrichment culture and bacteria were dominant. DGGE banding pattern revealed that archaeal community profile of the enrichment culture was different from that of the activated sludge used as an inoculum for the enrichment culture. The most dominant DGGE band of the enrichment culture was affiliated with Methanococci. Further research is necessary to investigate metabolic role of the dominant archaeal population to better understand microbial community in the perchlorate-reducing enrichment culture.

Biodegradation of PCBs into Contaminated Dredged Sediments (오염퇴적물에 함유된 PCBs의 생물분해 특성)

  • 안재환;김소정;지재성;배우근;황병기
    • Journal of Soil and Groundwater Environment
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    • v.9 no.2
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    • pp.41-47
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    • 2004
  • This research in the degradation of polychlorinated biphenyls(PCB) has focussed on the use of experimental enrichment cultures to obtain PCB-deading communities and identification of PCB-degrading bacteria accor야ng to pure culture. During 180 days, enrichment culture was performed to obtain PCB-degrading bacteria and initial concentration was injected 1.6 ppm,0.7 ppm, respectively. After 180 days of enrichment culture, PCBs was removed 80-87% and 57-71%. Biodegradation of PCBs was studied according to dominated PCB-degrading bacteria. Biodegraddation of PCBs was 80% in initial concentration of PCBs for 20days, enrichment cultured PCB-degrading bacteria was isolated by pure culture and it was verified to Pseudoxanthomonas sp.

Biodegradiation of Benzoate by Pseudomonas sp. (Pseudomonas sp.에 의한Benzoate의 생분해)

  • 김교창;정준영
    • Journal of Food Hygiene and Safety
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    • v.11 no.3
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    • pp.165-170
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    • 1996
  • The biodegradation of high concentration of benzoate by enrichment culture with Pseudomonas sp. was investigated. During 50 days continuous culture, average of removal rate of benzoate and COD were 90% and 83%, respectively. And the enzymatic activity of catechol 2,3-dioxygenase was determined in the continuous culture but not Catechol 1,2-dioxygenase. On the other hand, Pseudomonas sp in the culture was investigated with SEM and the result was revealed that the cell shape was more demage according concentration of benzoate.

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Microbial Community Structure in Hexadecane- and Naphthalene-Enriched Gas Station Soil

  • Baek, Kyung-Hwa;Kim, Hee-Sik
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.651-657
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    • 2009
  • Shifts in the activity and diversity of microbes involved in aliphatic and aromatic hydrocarbon degradation in contaminated soil were investigated. Subsurface soil was collected from a gas station that had been abandoned since 1995 owing to ground subsidence. The total petroleum hydrocarbon content of the sample was approximately 2,100 mg/kg, and that of the soil below a gas pump was over 23,000 mg/kg. Enrichment cultures were grown in mineral medium that contained hexadecane (H) or naphthalene (N) at a concentration of 200 mg/l. In the Henrichment culture, a real-time PCR assay revealed that the 16S rRNA gene copy number increased from $1.2{\times}10^5$to $8.6{\times}10^6$with no lag phase, representing an approximately 70-fold increase. In the N-enrichment culture, the 16S rRNA copy number increased about 13-fold after 48 h, from $6.3{\times}10^4$to $8.3{\times}10^5$. Microbial communities in the enrichment cultures were studied by denaturing gradient gel electrophoresis and by analysis of 16S rRNA gene libraries. Before the addition of hydrocarbons, the gas station soil contained primarily Alpha- and Gammaproteobacteria. During growth in the H-enrichment culture, the contribution of Bacteriodetes to the microbial community increased significantly. On the other hand, during N-enrichment, the Betaproteobacteria population increased conspicuously. These results suggest that specific phylotypes of bacteria were associated with the degradation of each hydrocarbon.

Comparison of Real-Time PCR and Conventional Culture Method for Detection of Cronobacter spp. in Powdered Foods (분말식품에서 Cronobacter spp. 검출을 위한 Real-Time PCR과 배지배양법의 비교검증)

  • Chon, Jung-Whan;Song, Kwang-Young;Kim, Sun-Young;Hyeon, Ji-Yeon;Kim, Yun-Gyeong;Hwang, In-Gyun;Kwak, Hyo-Sun;Seo, Kun-Ho
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.87-91
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    • 2011
  • The aim of this study was to compare the performance of conventional culture and real-time PCR for detection of Cronobacter spp. in powdered foods. Infant formula, baby food and Misugaru inoculated with Cronobacter were enriched in distilled water as first enrichment step, followed by incubating in Enterobacteriaceae enrichment (EE) broth as second enrichment step. A loopful of enriched sample was streaked onto Druggan-Forsythe-Iversen agar, followed by incubating at $37^{\circ}C$ for 24 h. One milliliter of the enriched distilled water and EE broth were used in real-time PCR assay. No statistical differences were observed in the number of positive samples between culture method and real-time PCR (p>0.05) in all types of food samples. The number of positives of real-time PCR was higher in the first enrichment media (distilled water) than the second enrichment media (EE broth), though there was no significant difference (p>0.05). It appears that some components of the second enrichment broth, EE broth, inhibit the reaction of real-time PCR. These results show that real-time PCR using a single enrichment with distilled water could be useful as an effective screening method for detection of Cronobacter while saving much time and labor compared to conventional culture method.

혐기성 PCE 탈염소화 미생물 농화 배양 및 미생물 군집 해석

  • 문부영;이태호;박태주
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.332-336
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    • 2004
  • An anaerobic PCE(tetrachloroethylene) dechlorinating bacterial culture from a landfill soil was enriched and characterized. The enrichment culture could dechlorinate 60$\mu$mol/$m\ell$ of PCE during a month of incubation and cis-DCE(cis-dichloroethylene) was observed as a main product of PCE dechlorination. Microbial analysis of the dechlorinating enrichment culture by rising PCR-DGGE (Polymerase chain reaction-Denaturing gradient gel electrophoresis) method showed that at least three microorganisms were related to the anaerobic PCE dechlorination.

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Anaerobic dechlorinating enrichment culture on tetrachloroethene (PCE) (PCE 탈염소화를 위한 혐기성배양)

  • Kim, Byung-Hyuk;Baek, Kyung-Hwa;Sung, Youl-Boong;Choi, Gang-Kook;Cho, Dae-Hyun;Oh, Hee-Mock;Kim, Hee-Sik
    • Proceedings of KOSOMES biannual meeting
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    • 2007.11a
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    • pp.185-185
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    • 2007
  • Starting at the beginning q the 20th century, increasing amounts of tetrach1oroethene (PCE) and trichloroethene (TCE)were manufactured due to the extensive use of these compounds in industry, in the military, and in private households, mainly as nonflammable solvents. This widespread use, along with careless handling and storage, are among the most serious contaminants of soil, sediment and groundwater. Highly chlorinated ethenes are typically not degraded through oxygenation by aerobic bacteria Since complete reductive dechlorination of PCE and TCE to ethene (ETH) has been observed in anaerobic enrichment culture, anaerobic dehalorespiring bacteria have received increased attention in the last decade. Under anaerobic conditions, these compounds con be reductively dehalogenated to less-chlorinated ethenes or innocuous ethene by microorganism through dehalorespiration. We have been studying anaerobic enrichment culture which used lactate as the electron donor for reductive dechlorination of PCE to ETH the anaerobic mixed microbial culture was enriched from the sediment sample taken from site contaminated with PCE. PCE was consistently and completely converted to ethene. In addition, the accumulation of intermediate products such as 1,2-ds-dichloroethene (cis-DCE) and vinyl chloride (VC) was observed in the anaerobic mixed microbial culture. the established dechlorinating enrichment culture was analyzed by DGGE using primers specific to DefrJ1ococcoides 16S rRNA gene sequences. In conclusion, we established the PCE dechlorinating enrichment culture and confirmed the existence of Dehalococcoides in an enrichment culture.

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