• Title/Summary/Keyword: microbiological monitoring

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Construction of a Bioluminescent Reporter Using the luc Gene and meta-Cleavage Dioxygenase Promoter for Detection of Catecholic Compounds

  • Park, Sang-Ho;Lee, Dong-Hun;Oh, Kye-Heon;Kim, Chi-Kyung
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.183-186
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    • 2000
  • Several types of bioluminescent reporter strains have been developed for the detection and monitoring of pollutant aromatics contaminating the environment. In this study, a bioluminescent reporter strain, E. coli SHP3, was constructed by fusing the luc gene of firefly luciferase with the promoter of pcbC responsible for the meta-cleavage of aromatic hydrocarbons. the bioluminescence expressed by the luc gene in the reporter was well triggered by the promoter when it was exposed to 2,3-dihydroxybiphenyI (2,3-DHBP) at 0.5 to 1 mM concentrations. The bioluminescent response was more extensive when the reporter strain was exposed to 5 mM catechol and 2 mM 4-chlorocatechol. These different types of bioluminescent responses by E. coli SHP3 appeared to be characterized by the nature of the aromatics to stress. Since E. coli SHP3 responded to 2,3-DHBP quite sensitively, this reporter strain could be applied for detecting some catecholic pollutants.

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Understanding of Extracellular Fumarate Induced dctA Gene Expression Profile Using GFP Reporter (GFP 리포터를 이용한 외부 푸마르산 유도 dctA 유전자 발현 특성 파악)

  • Irisappan, Ganesh;Ravikumar, Sambandam;Kim, Joo-Han;Hong, Soon-Ho
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.174-178
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    • 2011
  • In Escherichia coli, DcuS/R two-component system controls fumarate import and utilization related gene expression. To understand the dynamic response of the bacterium DcuS/R two-component system with respect to fumarate concentrations, DcuS/R induced dctA promoter was integrated with GFP reporter protein. Expression monitoring study using recombinant strain showed that dctA promoter was upregulated with 1 mM of fumarate in M9 minimal medium.

CORROSION OF STEEL GAS PIPELINE INDUCED BY SULFATE-REDUCING BACTERIA IN ANAEROBIC SOIL (혐기성 토양에 서식하는 황산염환원세균에 의한 가스배관의 미생물부식)

  • Li SeonYeob;Jeon KyungSoo;Kho YoungTai;Kang Tak
    • 한국가스학회:학술대회논문집
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    • 2001.10a
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    • pp.58-68
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    • 2001
  • Microbiologically influenced corrosion (MIC) of carbon steel gas pipeline in soil environments was investigated at field and laboratory MIC is very severe corrosion and it is not easy to distinguish this corrosion from Inorganic corrosion because of its localized, pitting-type character Therefore, it is important to provide proper assessment techniques for the prediction, detection, monitoring and mitigation of MIC. It is possible to predict the MIC risk, i.e., the activity of sulfate-reducing bacteria (SRB) through the analysis of soil environments. Chemical, microbiological and surface analysis of corrosion products and metal attacked could reveal the possibility of the occurrence of MIC. Various electrochemical and surface analysis techniques could be used for the study of MIC. Among these techniques, thin-film electrical resistance (ER) type sensors are promising to obtain localized corrosion rate of MIC induced by SRB. It is also important to study the effect of cathodic protection (CP) on the MIC In case of coated pipeline, the relationship between coating disbondment and the activity of SRB beneath the disbanded coating is also important.

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Assessment of Natural Attenuation Processes in the Groundwater Contaminated with Trichloroethylene (TCE) Using Multi-Species Reactive Transport Modeling (다성분 반응 이동 모델링을 이용한 트리클로로에틸렌(TCE)으로 오염된 지하수에서의 자연저감 평가)

  • Jeen, Sung-Wook;Jun, Seong-Chun;Kim, Rak-Hyeon;Hwang, Hyoun-Tae
    • Journal of Soil and Groundwater Environment
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    • v.21 no.6
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    • pp.101-113
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    • 2016
  • To properly manage and remediate groundwater contaminated with chlorinated hydrocarbons such as trichloroethylene (TCE), it is necessary to assess natural attenuation processes of contaminants in the aquifer along with investigation of contamination history and aquifer characterization. This study evaluated natural attenuation processes of TCE at an industrial site in Korea by delineating hydrogeochemical characteristics along the flow path of contaminated groundwater, by calculating reaction rate constants for TCE and its degradation products, and by using geochemical and reactive transport modeling. The monitoring data showed that TCE tended to be transformed to cis-1,2-dichloroethene (cis-1,2-DCE) and further to vinyl chloride (VC) via microbial reductive dechlorination, although the degree was not too significant. According to our modeling results, the temporal and spatial distribution of the TCE plume suggested the dominant role of biodegradation in attenuation processes. This study can provide a useful method for assessing natural attenuation processes in the aquifer contaminated with chlorinated hydrocarbons and can be applied to other sites with similar hydrological, microbiological, and geochemical settings.

Development of a Competitive Direct Enzyme-Linked Immunosorbent Assay for Teicoplanin

  • Lee, Hyang-Burm;Kwak, Bo-Yeon;Lee, Jae-Chan;Kim, Chang-Jin;Shon, Dong-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.612-619
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    • 2004
  • A competitive direct enzyme-linked immunosorbent assay (cdELISA) was developed for selective and rapid detection of a glycopeptide antibiotic, teicoplanin (TP). TP was conjugated to bovine serum albumin (BSA) for use as an immunogen. Repeated subcutaneous injections of 0.5 mg of the conjugate was effective in generating specific polyclonal antibody (PAb) toward TP in rabbits, as determined by cdELISA. TP-horseradish peroxidase conjugate (TP-HRP) was used as an enzyme marker. The cdELISA was developed based on a competition reaction between TP-BSA PAb and TP-HRP conjugate. The TP-BSA PAb was highly sensitive (detection limit, 0.3 ng/ml and specific toward teicoplanin, showing no cross-reactivity to other glycopeptide antibiotics including vancomycin. There were good correlations ($r^2$=0.84 and 0.76, respectively) between cdELISA and microbiological assay, and high-performance liquid chromatography. The cdELISA system developed in this work is expected to be useful not only for selective and rapid monitoring of TP but also for study of TP pharmacokinetics.

A NEWLY DEVELOPED CONTINUOUS TOXICITY TEST SYSTEM USING A LUMINOUSLY MODIFIED TERRESTRIAL BACTERIUM

  • Cho, Jang-Cheon;Lee, Kyu-Ho;Lee, Dong-Hun;Jahng, Deok-Jin;Park, Han-Oh;Kim, Sang-Jong
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.108-113
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    • 2000
  • Freshwater borne bacteria transformed with luxAB-containing plasmid were optimized for the toxicity tests of various organic carbons and heavy metals. The EC$\sub$50/ values obtained from tests using the most sensitive bacterium to toxicants, YH9-RC, revealed to be much less than those from the Microtox$\^$/. In addition, some physiological characteristics of this bacterium under the toxic stress conditions such as potential bioluminescence, specific growth rate, and intracellular ATP contents, reproducibly and reliably correlated to the toxicity of the chemicals exposed. The higher concentrations of COD in wastewater samples, the lower EC$\sub$50/ values, therefore the developed toxicity test was found to be easily applicable to the toxicity test for wastewater samples and effluents. The conditions for constructing 384-multiwell plate containing freeze-dried bacterium were also optimized through the addition of 0.16 M trehalose before freeze-drying. Consequently, the advanced test system featuring a continuous measurement of the toxicity, an automated real-time monitoring of its results, and an alerting function was designed and constructed in combination with the microbiological, mechanical, and electronic compartment.

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Comparison on serological reaction between complement fixation test and enzyme-linked immunosorbent assay for detection of antibodies against Sendai virus, mouse hepatitis virus and Mycoplasma pulmonis in mice and rats (마우스 및 랫트의 Sendai virus, mouse hepatitis Virus, Mycoplama pulmonis 감염(感染)에 대한 보체결합반응(補體結合反應)과 효소표식면역흡착측정법(酵素標識免疫吸着測定法)과의 비교(比較))

  • Chung, Yoo-yeul;Lee, Hak-cheul;Lee, Eun;Yoo, Byung-sam
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.517-523
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    • 1989
  • This study was undertaken to establish reliable diagnostic-procedures for the microbiological monitoring of laboratory animals. Murine(mice and rats) antibodies against hemagglutinating virus of Japan(HVJ), mouse hepatitis virus(MHV) and Mycoplasma pulmonis(Mp) were detected sensitively and specifically in experimentally and naturally infected animals' sera by an indirect enzyme-linked immunosorbent assay(ELISA), using urease conjugated antimurine immunoglobulin. The sensitivity and specificity of the complement fixation test which has been apllied widely for serodiagnosis of HVJ, MHV and Mp infections were apparently lower than those of ELISA. From these results, the ELISA was found to be available for the serodiagnosis of HVJ, MHV and Mp infections in mice and rats.

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Identification of Actinobacillus actinomycetemcomitans Using Species-Specific 16S rDNA Primers

  • Kim Su Gwan;Kim Soo Heung;Kim Mi Kwang;Kim Hwa Sook;Kook Joong Ki
    • Journal of Microbiology
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    • v.43 no.2
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    • pp.209-212
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    • 2005
  • The purpose of this study was to develop species-specific PCR primers for use in the identification and detection of Actinobacillus actinomycetemcomitans. These primers target variable regions of the 168 ribosomal RNA coding gene (rDNA). We assessed the specificity of the primers against 9 A. actinomycetemcomitans strains and 11 strains (3 species) of the Haemophilus genus. Primer sensitivity was determined by testing serial dilutions of the purified genomic DNAs of A. actinomycetemcomitans ATCC$ 33384^$T Our obtained data revealed that we had obtained species-specific amplicons for all of the tested A. actinomycetemcomitans strains, and that none of these amplicons occurred in any of the other species. Our PCR protocol proved able to detect as little as 4 fg of A. actinomycetemcomitans chromosomal DNA. Our findings suggest that these PCR primers are incredibly sensitive, and should prove suitable for application in epidemiological studies, as well as the diagnosis and monitoring of periodontal pathogens after treatment for periodontitis.

Improved Prediction of Coreceptor Usage and Phenotype of HIV-1 Based on Combined Features of V3 Loop Sequence Using Random Forest

  • Xu, Shungao;Huang, Xinxiang;Xu, Huaxi;Zhang, Chiyu
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.441-446
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    • 2007
  • HIV-1 coreceptor usage and phenotype mainly determined by V3 loop are associated with the disease progression of AIDS. Predicting HIV-1 coreceptor usage and phenotype facilitates the monitoring of R5-to-X4 switch and treatment decision-making. In this study, we employed random forest to predict HIV-1 biological phenotype, based on 37 random features of V3 loop. In comparison with PSSM method, our RF predictor obtained higher prediction accuracy (95.1% for coreceptor usage and 92.1% for phenotype), especially for non-B non-C HIV-l subtypes (96.6% for coreceptor usage and 95.3% for phenotype). The net charge, polarity of V3 loop and five V3 sites are seven most important features for predicting HIV-1 coreceptor usage or phenotype. Among these features, V3 polarity and four V3 sites (22, 12, 18 and 13) are first reported to have high contribution to HIV-1 biological phenotype prediction.

Testing a Small Scale Aseptic System for Milk in Plastic Bottles

  • Petrus, Rodrigo Rodrigues;Faria, Jose de Assis Fonseca
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.18-22
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    • 2007
  • The objective of this study was to develop and assess the performance of an aseptic system for liquid milk contained in plastic bottles, from a small-scale production standpoint. Commercial sterility tests conducted on the bottled milk were utilized in our assessments of the system, via the identification and monitoring of the principal points of the process. Four 150 L batches of milk with pH values of approximately 6.7 were heat-processed at between 137 and $143^{\circ}C$ for 10 see in a plate heat exchanger, and then aseptically transferred to 500 mL high-density polyethylene (HOPE) bottles, in an ISO class 7 clean room. The aseptic condition of the bottles was achieved via 10 see of rinsing with a mixture containing 0.5% peracetic acid and 0.8% hydrogen peroxide at $30^{\circ}C$, followed by another rinse with sterile water. Of the 4 batches processed, 2 were determined to exhibit commercial sterility, on the basis of the physical-chemical and microbiological criteria adopted. It was concluded that some adjustment of the processing line was required in order to achieve full commercial sterility for all processes. The aseptic system developed and assessed in this study was demonstrated to have great potential for the processing and transferring of milk into plastic bottles, from a small-scale production standpoint.