• Title/Summary/Keyword: genetic contamination

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Genetic Recombination by Protoplast Fusion of Cellulomonas sp CS 1-1 (원형질체 융합법에 의한 cellulomonas sp. CS 1-1의 유전자 재조합)

  • 김병홍;이학주
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.309-314
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    • 1985
  • Autoxotrophic mutants of a cellulolytic baterium Cellulomonas sp. CS 1-1 were grown at $30^{\circ}C$ for 6hr using a complete medium containing 0.5M sucrose and for another 90 min after addition of 0.3 U/ml penicillin G, and were protoplasted by 0.2mg/ml lysozyme for 2hr. Prototrophic recombinants were obtained at the rates of $10^{-3}$ to $10^{-5}$by fusing the protoplasts in the presence of 40% polyethyleneglycol3350. Nystatin could be used to eliminate fungal contamination during the regeneration of the plotaplasts.

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Genetic Diversity of Foodborne Pathogen Detected in Commercial Shellfish in Metropolitan Area

  • Park, Jung Hun;Cho, Kyu Bong
    • Biomedical Science Letters
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    • v.28 no.2
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    • pp.83-91
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    • 2022
  • This study was performed to investigate the contamination status of microorganisms that cause food poisoning in shellfish distributed in the metropolitan area of South Korea. The analyses were conducted according to the sample type, season, and region. In particular, the genotype of food poisoning viruses that explosively cause collective food poisoning was analyzed. Total 483 each of four types of shellfish (Crassostrea gigas, Mytilus coruscus, Pectea albicans albicans, and Scapharca subcrenata) were collected from four distribution sites located in the metropolitan area. We investigated foodborne pathogen by multiplex PCR and RT/Nested PCR from shellfish. As a result, Vibrio parahaemolyticus and Bacillus cereus were detected in 13 and 21 samples and Norovirus (NoV) GI and GII were detected in 7 and 12 samples, respectively. V. parahaemolyticus and NoV GI and GII showed differences in types of shellfish and seasons, and no correlation was confirmed with regional differences. Also, as a result of genotyping for the detected NoV GI and GII, they were confirmed as NoV GI.7, GI.5 and GII.3, which are causes of collective food poisoning. Therefore, cross-infection with shellfish can cause food poisoning. In particular, attention must be paid to the handling and cooking of shellfish in organizations that implement group feeding, and it is necessary to establish a management system for microorganisms that cause food poisoning in the process of shellfish farming and distribution.

Unexpected Presence of Blastocystis Subtype 1-3 DNA in Human Vaginal and Sperm Samples Coinfected with Trichomonas vaginalis

  • Villalobos, Guiehdani;Sanchez-Aguillon, Fabiola;Carmona-Maldonado, Marcia Veronica;Gonzalez-Arenas, Nelly Raquel;Lopez-Escamilla, Eduardo;Hernandez-Castro, Rigoberto;Romero-Valdovinos, Mirza;Martinez-Flores, Williams Arony;Ramirez-Hinojosa, Juan Pablo;Maravilla, Pablo;Martinez-Hernandez, Fernando
    • Parasites, Hosts and Diseases
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    • v.60 no.3
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    • pp.195-200
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    • 2022
  • There have been few reports on extra-enteric infections by Blastocystis STs and none have been molecularly identified in samples from human reproductive organs. We report for the first time the identification of 3 different subtypes of Blastocystis (ST1-3) in vaginal and sperm samples, from patients infected with Trichomonas vaginalis. Blastocystis STs were identified by PCR-sequencing and by phylogenetic inferences using 28 vaginal swab samples and 7 sperm samples from patients trichomoniasis. Blastocystis STs were identified in 6 of 28 vaginal swabs (21.4%) and in 3 of 7 sperm samples (42.8%). In both biological samples, STs 1-3 were found; one vaginal sample showed subtype co-infection with ST1 and ST3. High genetic variation was observed in the sequences obtained and no specific clustering in the phylogenetic trees was detected. Most of the haplotypes identified were placed far from the main dispersal centers. Our finding suggested that incorrect cleaning of the genital area or a contamination by combination of anal and vaginal intercourse.

Arsenic Detoxification by As(III)-Oxidizing Bacteria: A Proposition for Sustainable Environmental Management

  • Shamayita Basu;Samir Kumar Mukherjee;Sk Tofajjen Hossain
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.1-9
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    • 2023
  • Arsenic (As), which is ubiquitous throughout the environment, represents a major environmental threat at higher concentration and poses a global public health concern in certain geographic areas. Most of the conventional arsenic remediation techniques that are currently in use have certain limitations. This situation necessitates a potential remediation strategy, and in this regard bioremediation technology is increasingly important. Being the oldest representativse of life on Earth, microbes have developed various strategies to cope with hostile environments containing different toxic metals or metalloids including As. Such conditions prompted the evolution of numerous genetic systems that have enabled many microbes to utilize this metalloid in their metabolic activities. Therefore, within a certain scope bacterial isolates could be helpful for sustainable management of As-contamination. Research interest in microbial As(III) oxidation has increased recently, as oxidation of As(III) to less hazardous As(V) is viewed as a strategy to ameliorate its adverse impact. In this review, the novelty of As(III) oxidation is highlighted and the implication of As(III)-oxidizing microbes in environmental management and their prospects are also discussed. Moreover, future exploitation of As(III)-oxidizing bacteria, as potential plant growth-promoting bacteria, may add agronomic importance to their widespread utilization in managing soil quality and yield output of major field crops, in addition to reducing As accumulation and toxicity in crops.

Development of HRM Markers for Discrimination of Pyogo (Lentinula edodes) Cultivars Sanjo 701 and Chamaram

  • Suyun Moon;Hojin Ryu
    • The Korean Journal of Mycology
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    • v.50 no.3
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    • pp.225-233
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    • 2022
  • Pyogo (Shiitake, Lentinula edodes) is one of the most important edible mushrooms because of its outstanding nutritive and medicinal value. In the registration and protection procedure for newly developed mushroom cultivars, the application of molecular markers that can supplement the morphological characteristic-based distinction has been strongly requested. Sanjo 701 and Chamaram, newly developed at the Federation Forest Mushroom Research Center of Korea, have been characterized as innovative cultivars suitable for customer demands because of their high yields and cultivation rates. However, no technical tools can protect the rights to these important cultivars. In this study, using comparative genomic information from 23 commercially available pyogo cultivars, we identified single nucleotide polymorphisms (SNPs) that accurately differentiated Sanjo701 and Chamaram from the other cultivars. We also developed high-resolution melting analysis (HRM)-based SNP markers that discriminate among the tested 23 pyogo cultivars. The developed SNP markers can be utilized for rapid, accurate identification of pyogo cultivars with low genetic diversity and to prevent cultivar contamination caused by illegally distributed inocula. In addition, these markers can serve as a crucial scientific basis for securing the right to conserve new cultivars in international markets.

Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.88-89
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    • 2013
  • A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

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Evaluation of Biological Critical Control Points Using Escherichia coli Genotyping (Escherichia coli Genotype을 이용한 생물학적 Critical Control Point의 적합성 평가)

  • Kim, Hak-Jae;Hahn, Tae-Wook;Juong, Ji-Hun;Bahk, Gyung-Jin;Hong, Chong-Hae
    • Food Science of Animal Resources
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    • v.29 no.6
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    • pp.695-701
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    • 2009
  • This study was performed to evaluate the effectiveness of biological critical control points using the genetic profile of Escherichia coli isolates from pork cutting plants. Samples were collected from carcasses, equipment (knife, table, glove, transport belt, boning and skinning machine), the environment (wall and floor), and meat cuts during the cutting process from two plants. Pulsed-field gel electrophoresis (PFGE) was used to characterize the E. coli isolates. An identical genotype was detected from the carcasses, equipment, environment, and final meat cuts, and showed that the incoming carcasses, which were contaminated during transportation from slaughterhouses, were a major source of E. coli that was spread throughout processing. Also, consistent cross-contamination due to improper cleaning and disinfection procedures was another possibility. As a result, incoming carcasses and cleaning procedures should be considered critical control points in pork cutting plants, since a heating step is not used to inactivate microorganisms. Furthermore, the high rate (59.6%) of E. coli isolation indicates E. coli can be a good indicator in livestock processing plants even though it has genetic diversity.

An Overview of Geoenvironmental Implications of Mineral Deposits in Korea (한반도 광상 성인유형에 따른 환경 특성)

  • 최선규;박상준;이평구;김창성
    • Economic and Environmental Geology
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    • v.37 no.1
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    • pp.1-19
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    • 2004
  • Metallic deposits in Korea have a variety of genetic types such as hydrothermal veins, skarns, hydrothermal replacement and alaskite deposits and so on. Geological, mineralogical and geochemical features including host rock, wall-rock alteration, ore and gangue mineralogy, mineral texture and secondary mineralogy related to weathering process control the environmental signatures of mining areas. The environmental signatures of metallic deposits closed from early 1970s to late 1990s in Korea show complicate geochemistry and mineralogy due to step weathering of primary and secondary minerals such as oxidation-precipitation-remobilization. The potentiality of low pH and high heavy metal Concentration s from acid mine drainage is great in base-metal deposits associated with polymetallic mineralization, breccia-pipe type and Cretaceous hydrothermal Au veins with the amount of pyrite whereas skam, hydrothermal replacement, hydrothermal Cu and Au-Ag vein deposits are in low contamination possibility. The geoenvironmental models reflecting the various geologic features closely relate to disuibution of sulfides and carbonates and their ratios and finally effect on characteristics of environmental signatures such as heavy metal species and their concentrations in acid mine drainage.

Polymorphism of inter simple sequence repeat markers in Hypsizygus marmoreus (Inter Simple Sequence Repeat(ISSR) 마커를 활용한 느티만가닥버섯(Hypsizigus marmoreus) 종내 다형성 분석)

  • Oh, Youn-Lee;Nam, Yun-Geul;Jang, Kab-Yeul;Kong, Won-Sik;Oh, Min ji;Im, Ji-Hoon;Choi, In-Geol
    • Journal of Mushroom
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    • v.15 no.4
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    • pp.273-278
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    • 2017
  • Hypsizygus marmoreus is a mushroom with abundant flavor and medicinal properties. However, its application is limited by problems such as long cultivation period, low biological efficiency, and microbiological contamination; therefore, there is a substantial need for development of new cultivars of this species. In this study, 55 strains of H. marmoreus were subjected to inter simple sequence repeat (ISSR) analysis to identify markers for the selection of mother strains for breeding from the collected germplasm. ISSR 13 and 15 were confirmed as polymorphic markers. The three strains (KMCC03106, KMCC03107, and KMCC03108) with white cap color were found to be genetically closely related upon UPGMA analysis of both ISSR 13 and 15. Based on the PCR analysis results for ISSR 15, the collected germplasm were differentiated into three groups according to the strain collection year. Thus, ISSR 15 could be a marker for determining the phylogeny of cap color and genetic variations according to the strain collection year. These results suggest that ISSR markers can be effective tools for the selection of mother strains for breeding of H. marmoreus.

Molecular Epidemiology of Legionella pneumophila Isolated from Water Supply Systems in Seoul, Korea (서울시 수계시설에서 분리된 Legionella pneumophila의 분자역학적 특성)

  • Jeon, Su Jin;Jung, Ji Hun;Seung, Hyun Jung;Kim, Chang Kyu;Jin, Young Hee;Oh, Young Hee;Choi, Sung Min;Chae, Young Zoo
    • Journal of Environmental Health Sciences
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    • v.39 no.2
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    • pp.166-177
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    • 2013
  • Objectives: The genus Legionella is common in aquatic environments. Some species of Legionella are recognized as potential opportunistic pathogens for human, notably Legionella pneumophila that causes, Legionellosis. Thus, we investigated the contamination of Legionella pneumophila on water supply systems in Seoul, including cooling towers, public baths, hospitals and fountains. Methods: The existence of 16S rRNA and mip gene of L. pneumophila was confirmed in the genome of the isolated strains by PCR. Results: During the summer season of 2010 and 2011, Legionella pneumophila were detected from 163 samples (21.1%) out of 772 samples collected. Among the 163 strains of L. pneumophila, eighty one isolates belonged to serogroup 1 (57.4%), 23 isolates were serogroup 5 (16.3%), 21 isolates were serogroup 6 (14.9%), 8 isolates were serogroup 2 (5.79%), and 8 isolates were identified in serogroup 3 (5.7%). Through PFGE (pulsed-field gel electrophoresis) analysis using Sfi I, genetic types of L. pneumophila were classified into five (A to E) patterns by the band similarity with excess of 70% from public baths. Conclusions: The PFGE patterns of the serotypes showed a tendency for diversity of L. pneumophila. Our results suggest the existence of serological and genetic diversity among the L. pneumophila isolates.