• Title/Summary/Keyword: Microbial culture

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Microbial Basis for Enhanced Degradation of the Fumigant 1,3-Dichloropropene (1,3-D) in Soil

  • Chung, Keun-Yook
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.10a
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    • pp.125-139
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    • 2000
  • The differential enhanced degradation of cis- and trans-1,3-D was observed in the previous two studies performed by Ou et al. (1995) and especially Chung et al. (1999). This study was initiated to investigate the involvement of microorganisms in the differential enhanced degradation of the chemicals. As expected, microorganisms were responsible for the enhanced degradation of the chemicals. A mixed bacterial culture capable of degrading 1,3-D was isolated from an enhanced soil sample collected from a site treated with 1,3-D. Similar to the enhanced soil, the mixed culture degraded trans-1,3-D faster than cis-1,3-D. This mixed culture could not utilize cis- and trans-1,3-D as a sole source of carbon for growth. Rather, a variety of second substrates were evaluated to stimulate the differential enhanced degradation of the two isomers. As a result, the mixed culture degraded cis- and trans-1,3-D only in the presence of a suitable second substrate. Second substrates that had the capacity to stimulate the degradation included soil leachate, tryptone, tryptophan, and alanine. Other substrates tested, including soil extract, glucose, yeast extract, and indole (ailed to stimulate the degradation of the two isomers. Therefore, it appeared that the degradation of cis- and trans-1,3-D was a cometabolic process. The mixed culture was composed of four morphologically distinctive bacterial colonies.

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Tissue culture of medicinal plants: micropropagation, transformation and production of useful secondary metabolites

  • Yoshimatsu, Kayo
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.88-94
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    • 2005
  • Plant tissue culture studies have been done for the preservation of medicinal plant resources and efficient production of pharmaceutically important secondary metabolites. Micropropagation methods for Cephaelis ipecacuanha have been established and these methods enabled much more efficient propagation of the plants than the conventional methods using seedling or layering. The C. ipecacuanha plants derived from tissue culture grew uniformly in the field and they showed higher alkaloid contents compared to the plants grown from seedlings. Hairy root cultures of C. ipecacuanha and Panax ginseng have been established by infection with Agrobacterium rhizogenes, and the production of important pharmaceuticals by these cultures have been successfully demonstrated. In the case of C. ipecacuanha, the highest alkaloid yields from the hairy roots cultured for 8 weeks were 2.75-fold cephaeline (5.5 mg) and one third emetine (0.7 mg) compared with those from the roots of one-year old plant propagated through shoot-tip culture and cultivated in a greenhouse (2.0 mg cephaeline and 2.0 mg emetine). In the case of P. ginseng, ginsenoside contents in the hairy roots optimally cultured for 4 weeks were much higher than those in the roots of 4-year old field-grown plant. Thus our medicinal plant tissue cultures demonstrate desirable properties. However, they are always exposed to danger of microbial contamination or unexpected trouble of culture facilities. Cryopreservation of plant tissue cultures is a reliable method for long-term preservation. Cryopreservation studies on these cultures are also presented.

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Genetic characterization of Phellinus baumii PMO-P4 by analyzing restriction fragment length polymorphisms of nuclear ribosomal DNA internal transcribed spacers (ITS) (Ribosomal DNA의 ITS부위에 대한 RFLP 분석에 의한 Phellinus baumii PMO-P4의 유전학적 특성)

  • Chang, Yun-Hee;Kim, Tae-Rack;Kim, Hyun-Su;Yeo, Ik-Hyun;Lee, Sang-Youn;Ha, Hyo-Cheol
    • Journal of Mushroom
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    • v.4 no.2
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    • pp.43-47
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    • 2006
  • PMO-P4, being cultivated as "Sanghwang" in Korea, was proved to be P. baumii based on ITS (internal transcribed spacer) sequencing and RFLP (Restriction Fragment Length Polymorphism) patterns along with some Phellinus species including P. linteus. The similaraty of ITS sequencing between PMO-P4 and other Phellinus species was given the range of 48.6%~72.2%, showing the highest homology from P. linteus and the lowest from P. gilvus.

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Analysis of Pathogenic Microorganism's Contamination on Cultivation Environment of Strawberry and Tomato in Korea

  • Oh, Soh-Young;Nam, Ki-Woong;Kim, Won-Il;Lee, Mun Haeng;Yoon, Deok-Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.47 no.6
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    • pp.510-517
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    • 2014
  • The purpose of this study was to analyze microbial hazards for cultivation environments and personal hygiene of strawberry and tomato farms at the growth and harvesting stage. Samples were collected from thirty strawberry farms and forty tomato farms located in Korea and tested for Staphylococcus aureus and Bacillus cereus. To investigate the change in the distribution of the S. aureus and B. cereus, a total of 4,284 samples including air born, soil or medium, mulching film, harvest basket, groves and irrigation water etc. were collected from eight strawberry farms and nine tomato farms for one year. As a result, total S. aureus and B. cereus in all samples were detected. Among the total bacteria of strawberry farms, S. aureus (glove: $0{\sim}2.1Log\;CFU/100cm^2$, harvest basket: $0{\sim}3.0Log\;CFU/100cm^2$, soil or culture media: 0~4.1 Log CFU/g, mulching film: $0{\sim}3.8Log\;CFU/100cm^2$), B. cereus (glove: $0{\sim}2.8Log\;CFU/100cm^2$, harvest basket: $0{\sim}4.8Log\;CFU/100cm^2$, soil or culture media: 0~5.3 Log CFU/g, mulching film: $0{\sim}4.5Log\;CFU/100cm^2$) were detected in all samples. The total bacteria of tomato farms, S. aureus (glove: $0{\sim}4.0Log\;CFU/100cm^2$, harvest basket: $0{\sim}5.0Log\;CFU/100cm^2$, soil or culture media: 0~6.1 Log CFU/g, mulching film: $0{\sim}4.0Log\;CFU/100cm^2$), B. cereus (glove: $0{\sim}4.0Log\;CFU/100cm^2$, harvest basket: $0{\sim}4.3Log\;CFU/100cm^2$, soil or culture media: 0~5.9 Log CFU/g, mulching film: $0{\sim}4.7Log\;CFU/100cm^2$) were detected in all samples. The contamination of S. aureus and B. cereus were detected in soil, mulching film and harvest basket from planting until harvest to processing, with the highest count recorded from the soil. But S. aureus and B. cereus were not detected in irrigation water samples. The incidence of S. aureus and B. cereus in hydroponics culture farm were less than those in soil culture. The amount of S. aureus and B. cereus detected in strawberry and tomato farms were less than the minimum amount required to produce a toxin that induces food poisoning. In this way, the degree of contamination of food poisoning bacteria was lower in the production environment of the Korea strawberry and tomato, but problems can be caused by post-harvest management method. These results will be used as fundamental data to create a manual for sanitary agricultural environment management, and post-harvest management should be performed to reduce the contamination of hazardous microorganisms.

Monitoring Anaerobic Reductive Dechlorination of TCE by Biofilm-Type Culture in Continuous-Flow System (연속흐름반응조에서 바이오필름형태의 탈염소화 미생물에 의한 TCE분해 모니터링)

  • Park, Sunhwa;Han, Kyungjin;Hong, Uijeon;Ahn, Hongil;Kim, Namhee;Kim, Hyunkoo;Kim, Taeseung;Kim, Young
    • Journal of Soil and Groundwater Environment
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    • v.17 no.5
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    • pp.49-55
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    • 2012
  • A 1.28 L-batch reactor and continuous-flow stirred tank reactor (CFSTR) fed with formate and trichloroethene (TCE) were operated for 120 days and 56 days, respectively, to study the effect of formate as electron donor on anaerobic reductive dechlorination (ARD) of TCE to cis-1,2-dichloroethylene (c-DCE), vinyl chloride (VC), and ethylene (ETH). In batch reactor, injected 60 ${\mu}mol$ TCE was completely degraded in the presence of 20% hydrogen gas ($H_2$) in less than 8 days by anaerobic dechlorination mixed-culture (300 mg-soluble protein), Evanite Culture with ability to completely degrade tetrachloroethene (PCE) and -TCE to ETH under anaerobic conditions. Once the formate was used as electron donor instead of hydrogen gas in batch or chemostat system, the TCE-dechlorination rate decreased and acetate production rate increased. It indicates that the concentration of hydrogen produced in both systems is possibly more close to threshold for homoacetogenesis process. Soluble protein concentration of Evanite culture during the batch test increased from 300 mg to 688 mg for 120 days. Through the protein monitoring, we confirmed an increase of microbial population during the reactor operation. In CFSTR test, TCE was fed continuously at 9.9 ppm (75.38 ${\mu}mol/L$) and the influent formate feed concentration increased stepwise from 1.3 mmol/L to 14.3 mmol/L. Injected TCE was accumulated at 18 days of HRT, but TCE was completely degraded at 36 days of HRT without accumulation of the injected-TCE during the left of experiment period, getting $H_2$ from fermentative hydrogen production of injected formate. Although c-DCE was also accumulated for 23 days after beginning of CFSTR operation, it reached steady-state in the presence of excessive formate. We also evaluated microbial dynamic of the culture at different chemical state in the reactor by DGGE (denaturing gradient gel electrophoresis).

Development of a Screening Method and Device for the Detection of Escherichia coli from Agri-Food Production Environments and Fresh Produce

  • Yun, Bohyun;An, Hyun-Mi;Shim, Won-Bo;Kim, Won-Il;Hung, Nguyen Bao;Han, Sanghyun;Kim, Hyun-Ju;Lee, Seungdon;Kim, Se-Ri
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2141-2150
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    • 2017
  • This study was conducted to develop a screening method using Colilert-18 and a device for the detection of E. coli from agri-food production environments and fresh vegetables. The specificity and sensitivity of Colilert-18 by temperature ($37^{\circ}C$ and $44^{\circ}C$) were evaluated with 38 E. coli and 78 non-E. coli strains. The false-positive rate was 3.8% (3/78) and 0% (0/78) at $37^{\circ}C$ and $44^{\circ}C$, respectively. The detection limit of E. coli at $37^{\circ}C$ at <1.0 log CFU/250 ml was lower than that at $44^{\circ}C$. The efficiency of the developed device, which comprised an incubator equipped with a UV lamp to detect E. coli in the field, was evaluated by measuring the temperature and UV lamp brightness. The difference between the set temperature and actual temperature of the developed device was about $1.0^{\circ}C$. When applying the developed method and device to various samples, including utensils, gloves, irrigation water, seeds, and vegetables, there were no differences in detection rates of E. coli compared with the Korean Food Code method. For sanitary disposal of culture samples after experiments, the sterilization effect of sodium dichloroisocyanurate (NaDCC) tablets was assessed for use as a substitute for an autoclave. The addition of one tablet of NaDCC per 50 ml was sufficient to kill E. coli cultured in Colilert-18. These results show that the developed protocol and device can efficiently detect E. coli from agri-food production environments and vegetables.

Microbial Communities of the Microbial Fuel Cell Using Swine Wastewater in the Enrichment Step with the Lapse of Time (가축분뇨를 이용한 미생물연료전지의 농화배양 단계에서 미생물 군집 변화)

  • Jang, Jae Kyung;Hong, Sun Hwa;Ryou, Youg Sun;Lee, Eun Young;Chang, In Seop;Kang, Young Koo;Kim, Jong Goo
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.12
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    • pp.973-977
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    • 2013
  • These studies were attempted to investigate the change of microbial community of anode of microbial fuel cell using swine wastewater in the enrichment step with the lapse of time. Microbial fuel cells enriched by a 1 : 1 mixture of anaerobic digestive juices of the sewage treatment plant and livestock wastewater. Enrichment culture step was divided into three stages to indentify the microorganisms. It was separated by each lag phase, exponential phase, and stationary phase. These steps were determined by the change of the current value. The current after enrichment was generated about $0.84{\pm}0.06mA$. We were cut out the different 17 bands in the DGGE fingerprint gel to do sequencing. The bands which the concentration was increasing or newly appearing with the lapse of time were included for this study. In the lag and exponential phase, Clostridium, Rhodocyclaceae, Bacteriodetes, and Uncultured bacterium etc. were detected. There were in the stationary phase Geobacter sp., Rhodocyclaceae, Candidatus, Nitrospira, Flavobactriaceae and uncultured bacterium etc. Geobactor among microorganisms detected in this study is known as the Electrochemically active microorganisms. It may include electrochemically active microorganisms to be considered as electrical activity microorganisms.

Effects of Nitrogen and Phosphorus Starvation on Growth and Fatty Acid Production in Newly Isolated Two Freshwater Green Microalgae from Nakdonggang River (낙동강 수계에서 분리한 녹조류 2종의 질소와 인의 결핍에 따른 생장 및 지방산 변화 연구)

  • Yim, Kyung June;Park, Hanwool;Lee, Chang Soo;Jo, Bok Yeon;Nam, Seung Won;Lee, Choul-Gyun;Kim, Z-Hun
    • Journal of Marine Bioscience and Biotechnology
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    • v.11 no.2
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    • pp.81-88
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    • 2019
  • In this study, effects of nitrogen (N) and phosphorus (P) starvation on the cell growth and fatty acid (FA) production of newly isolated freshwater microalgae were investigated. The microalgae were identified as Chlorella sp. and Parachlorella sp. through 18S rRNA sequencing. Optimal culture temperature and light intensity were investigated using a high-throughput photobioreator, and the result was validated in 0.5 L bubble column photobioreactors using BG-11 without NaNO3 and/or K2HPO4. Under nutrient starvation conditions, total FA contents of the microalgae were significantly changed rather than FA composition. Starvation of both N and P was most effective for increasing FA contents in Parachlorella sp (24.4±0.1%) whereas highest FA contents (42.6±1.8%) was achieved when only P was starved in Chlorella sp. among tested conditions. These results suggest an effective strategy for increasing FA production from microalgae using appropriate nutrient starvation.

Physical and Chemical Composition, Anti-Microbial Activity and Cytotoxicity of Kiwifruit Skin (식품표시 관련 교육경험에 따른 중학생들의 식품표시에 대한 인식과 활용실태)

  • Heo, Buk-Gu;Park, Yong-Seo;Im, Myung-Hee;Oh, Kyung-Taek;Cho, Ja-Yong
    • The Korean Journal of Community Living Science
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    • v.20 no.1
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    • pp.61-69
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    • 2009
  • This study was conducted to examine the chemical composition of kiwifruit skin, and to est its anti-microbial activities and cytotoxicities, thus, exploring ways for the economic utilization of kiwifruit skin. Four varieties of kiwifruit were examined: Daeheung, Bidan, Haegeum No.1 and Hayward. Vitamin C content in the fruit skins of Bidan, Daeheung, Haegeum No.1 and Hayward were 72.44, 67.22, 62.51 and 61.44mg/100g, respectively. Total amino acids content in the fruit skins of Bidan, Haegeum No.1, Hayward and Daeheung ere 808.31, 706.02, 629.07 and 464.83mg/100g dry weight, respectively. K and Ca content ere $17.20-45.70{\mu}g/mL$ and $4.58-10.15{\mu}g/mL$. While, other inorganic matter contents were below $4.89{\mu}g/mL$. Anti-microbial activity of kiwifruit skin extracts, in terms of the diameter of inhibition zone when tested against five gram positive and three gram negative microbial trains (even in the concentration of 2,000mg/L), was less than 14.1mm. The hyperplasia inhibition of lung cancer cells by methanol extracts from Bidan and Haegeum No.1 using concentrations of 800mg/L were 27.7% and 14.5%, however, those from Daeheung and Hayward were below 3% Consequently, it will be useful to know that kiwifruit skin can be added to processed goods which demand for higher concentrations of vitamin C, amino acids, K and Ca.

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Profiling Total Viable Bacteria in a Hemodialysis Water Treatment System

  • Chen, Lihua;Zhu, Xuan;Zhang, Menglu;Wang, Yuxin;Lv, Tianyu;Zhang, Shenghua;Yu, Xin
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.995-1004
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    • 2017
  • Culture-dependent methods, such as heterotrophic plate counting (HPC), are usually applied to evaluate the bacteriological quality of hemodialysis water. However, these methods cannot detect the uncultured or viable but non-culturable (VBNC) bacteria, both of which may be quantitatively predominant throughout the hemodialysis water treatment system. Therefore, propidium monoazide (PMA)-qPCR associated with HPC was used together to profile the distribution of the total viable bacteria in such a system. Moreover, high-throughput sequencing of 16S rRNA gene amplicons was utilized to analyze the microbial community structure and diversity. The HPC results indicated that the total bacterial counts conformed to the standards, yet the bacteria amounts were abruptly enhanced after carbon filter treatment. Nevertheless, the bacterial counts detected by PMA-qPCR, with the highest levels of $2.14{\times}10^7copies/100ml$ in softener water, were much higher than the corresponding HPC results, which demonstrated the occurrence of numerous uncultured or VBNC bacteria among the entire system before reverse osmosis (RO). In addition, the microbial community structure was very different and the diversity was enhanced after the carbon filter. Although the diversity was minimized after RO treatment, pathogens such as Escherichia could still be detected in the RO effluent. In general, both the amounts of bacteria and the complexity of microbial community in the hemodialysis water treatment system revealed by molecular approaches were much higher than by traditional method. These results suggested the higher health risk potential for hemodialysis patients from the up-to-standard water. The treatment process could also be optimized, based on the results of this study.