• Title/Summary/Keyword: culturable microorganisms

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Metagenome, the Untapped Microbial Genome, toward Discovery of Novel Microbial Resources and Application into the Plant Pathology

  • Lee, Seon-Woo
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.93-98
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    • 2005
  • Molecular ecological studies of microbial communities revealed that only tiny fraction of total microorganisms in nature have been identified and characterized, because the majority of them have not been cultivated. A concept, metagenome, represents the total microbial genome in natural ecosystem consisting of genomes from both culturable microorganisms and viable but non-culturable bacteria. The construction and screening of metagenomic libraries in culturable bacteria constitute a valuable resource for obtaining novel microbial genes and products. Several novel enzymes and antibiotics have been identified from the metagenomic approaches in many different microbial communities. Phenotypic analysis of the introduced unknown genes in culturable bacteria could be an important way for functional genomics of unculturable bacteria. However, estimation of the number of clones required to uncover the microbial diversity from various environments has been almost impossible due to the enormous microbial diversity and various microbial population structure. Massive construction of metagenomic libraries and development of high throughput screening technology should be necessary to obtain valuable microbial resources. This paper presents the recent progress in metagenomic studies including our results and potential of metagenomics in plant pathology and agriculture.

PCR-DGGE as a Supplemental Method Verifying Dominance of Culturable Microorganisms from Activated Sludge

  • Zhou, Sheng;Wei, Chaohai;Ke, Lin;Wu, Haizhen
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1592-1596
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    • 2010
  • To verify the dominance of microorganisms in wastewater biological treatment, PCR-DGGE (denaturing gradient gel electrophoresis) was performed as a supplementary support method for screening of the dominant microorganisms from activated sludge. Results suggest that the dominant microorganisms in activated sludge are primarily responsible for strengthening its effectiveness as a biological treatment system, followed by the non-main dominant microorganisms, whereas the non-dominant microorganisms showed no effects. The degree of microbial abundance present on the profile of PCR-DGGE was in line with the treatment efficiency of augmented activated sludge with isolated cultures, suggesting that PCR-DGGE can be used as an effective supplementary method for verifying culturable dominant microorganisms in activated sludge of coking wastewater.

Analysis techniques for fermented foods microbiome (발효식품의 마이크로바이옴 분석 기술)

  • Cha, In-Tae;Seo, Myung-ji
    • Food Science and Industry
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    • v.50 no.1
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    • pp.2-10
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    • 2017
  • Human have eaten various traditional fermented foods for a numbers of million years for health benefit as well as survival. The beneficial effects of fermented foods have been resulted from complex microbial communications within the fermented foods. Therefore, the holistic approaches for individual identification and complete microbial profiling involved in their communications have been of interest to food microbiology fields. Microbiome is the ecological community of microorganisms that literally share our environments including foods as well as human body. However, due to the limitation of culture-dependent methods such as simple isolations of just culturable microorganisms, the culture-independent methods have been consistently developed, resulting in new light on the diverse non-culturable and hitherto unknown microorganisms, and even microbial communities in the fermented foods. For the culture-independent approaches, the food microbiome has been deciphered by employing various molecular analysis tools such as fluorescence in situ hybridization, quantitative PCR, and denaturing gradient gel-electrophoresis. More recently, next-generation-sequencing (NGS) platform-based microbiome analysis has been of interest, because NGS is a powerful analytical tool capable of resolving the microbiome in respect to community structures, dynamics, and activities. In this overview, the development status of analysis tools for the fermented food microbiome is covered and research trend for NGS-based food microbiome analysis is also discussed.

Concentrations of total culturable microorganisms and Its Identification in Public Facilities (다중이용시설의 실내공기 중 총부유세균 농도와 종류)

  • Jeon, Byoung-Hak;Hwang, In Young
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.1
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    • pp.868-876
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    • 2015
  • This study was conducted to measure the concentrations of total culturable bacteria (TCB) and to identify the bacteria in thirty public facilities (7 elderly-care facilities, 4 hypermarkets, 4 university hospitals, 7 child care facilities, 4 subway stations and 4 bus terminals) in Seoul and Gyeonggi-Do Province. Although all concentrations measured in these study facilities did not exceed the national maintenance standard, it is believed that elderly-, child-care facilities should be high priority facilities to improve the indoor air quality and it is important to study the health effects and the airborne bacteria concentration of public facilities including non-culturable bacteria or allergen.

Identification and Phylogenetic Analysis of Culturable Bacteria in the Bioareosol from Several Environments (환경 유형에 따른 바이오에어로졸 중 배양성 세균 동정 및 계통분석)

  • Lee, Siwon;Chung, Hyen-Mi;Park, Su Jeong;Choe, Byeol;Kim, Ji Hye;Lee, Bo-Ram;Joo, Youn-Lee;Kwon, Oh Sang;Jheong, Weon Hwa
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.142-149
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    • 2015
  • Bioaerosols are comprised of particles 0.02-100 μm in size that originate in natural and artificial environments, and as a result of human activities. They consist of microorganisms including viruses, bacteria, fungi, and protozoa; fungal spores; microbial toxins; pollen; plant or animal material; expectorated liquid from humans; and glucans (peptidoglycan and β-glucan). Bioaerosols can cause respiratory and other diseases in humans and animals. In this study, bioaerosol samples acquired from agricultural sources, livestock, a sewage treatment plant, a beach, and a pristine area were analyzed to identify and phylogenetically characterize culturable microorganisms. The isolated bacteria exhibited regional differences, with different species dominating. However, Bacillus cereus was isolated in all samples, with a total of 31 strains isolated from all areas, and Acinetobacter baumannii was isolated from an indoor poultry farm. In addition, bacteria determined to be of novel genus or species of the genera Domibacillus, Chryceobacterium, Nocardioides and family Comamonadaceae were isolated from the agricultural, livestock and beach environments.

Identification of Culturable Bioaerosols Collected over Dryland in Northwest China: Observation using a Tethered Balloon

  • Chen, Bin;Kobayashi, Fumihisa;Yamada, Maromu;Kim, Yang-Hoon;Iwasaka, Yasunobu;Shi, Guang-Yu
    • Asian Journal of Atmospheric Environment
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    • v.5 no.3
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    • pp.172-180
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    • 2011
  • The transfer of microorganisms is important process for ecosystems. Microorganisms in dryland can transport itself to wetland through atmospheric diffusion, but only few papers reported about the atmospheric bioaerosol present over dryland. We carried out the direct sampling using a tethered balloon over Dunhuang City, China's northwestern dryland. Bioaerosols were collected using a tethered balloon with a bioaerosol collector at 820 m above the ground (1,960 m above the sea level) around noon on August 17, 2007. The bioaerosols were cultured after the collection at Dunhuang Meteorological observatory. Two strains of molds were isolated using the Nutrient agar medium. About 400-bp 18S rRNA partial sequences were amplified by PCR and determined afterwards. The results of a homology search by 18S rRNA sequences of isolates in DNA databases (GenBank, DDBJ, and EMBL) and an observation of the form revealed that two bioaerosols in the convective mixed layer over Dunhuang City were Cladosporium sp. and Aspergillus sp.

Bioprospecting Potential of the Soil Metagenome: Novel Enzymes and Bioactivities

  • Lee, Myung Hwan;Lee, Seon-Woo
    • Genomics & Informatics
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    • v.11 no.3
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    • pp.114-120
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    • 2013
  • The microbial diversity in soil ecosystems is higher than in any other microbial ecosystem. The majority of soil microorganisms has not been characterized, because the dominant members have not been readily culturable on standard cultivation media; therefore, the soil ecosystem is a great reservoir for the discovery of novel microbial enzymes and bioactivities. The soil metagenome, the collective microbial genome, could be cloned and sequenced directly from soils to search for novel microbial resources. This review summarizes the microbial diversity in soils and the efforts to search for microbial resources from the soil metagenome, with more emphasis on the potential of bioprospecting metagenomics and recent discoveries.

Application of Molecular Biology to Rumen Microbes -Review-

  • Kobayashi, Y.;Onodera, R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.77-83
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    • 1999
  • Molecular biological techniques that recently developed, have made it possible to realize some of new attempts in the research field of rumen microbiology. Those are 1) cloning of genes from rumen microorganisms mainly in E. coli, 2) transformation of rumen bacteria and 3) ecological analysis with nonculturing methods. Most of the cloned genes are for polysaccharidase enzymes such as endoglucanase, xylanase, amylase, chitinase and others, and the cloning rendered gene structural analyses by sequencing and also characterization of the translated products through easier purification. Electrotransformation of Butyrivibrio fibrisolvens and Prevotella ruminicola have been made toward the direction for obtaining more fibrolytic, acid-tolerant, depoisoning or essential amino acids-producing rumen bacterium. These primarily required stable and efficient gene transfer systems. Some vectors, constructed from native plasmids of rumen bacteria, are now available for successful gene introduction and expression in those rumen bacterial species. Probing and PCR-based methodologies have also been developed for detecting specific bacterial species and even strains. These are much due to accumulation of rRNA gene sequences of rumen microbes in databases. Although optimized analytical conditions are essential to reliable and reproducible estimation of the targeted microbes, the methods permit long term storage of frozen samples, providing us ease in analytical work as compared with a traditional method based on culturing. Moreover, the methods seem to be promissing for obtaining taxonomic and evolutionary information on all the rumen microbes, whether they are culturable or not.

An Overview of Genetic Information of Latent Mycobacterium tuberculosis

  • Hamidieh, Faezeh;Farnia, Parissa;Nowroozi, Jamileh;Farnia, Poopak;Velayati, Ali Akbar
    • Tuberculosis and Respiratory Diseases
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    • v.84 no.1
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    • pp.1-12
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    • 2021
  • Mycobacterium tuberculosis has infected more than two billion individuals worldwide, of whom 5%-10% have clinically active disease and 90%-95% remain in the latent stage with a reservoir of viable bacteria in the macrophages for extended periods of time. The tubercle bacilli at this stage are usually called dormant, non-viable, and/or non-culturable microorganisms. The patients with latent bacilli will not have clinical pictures and are not infectious. The infections in about 2%-23% of the patients with latent status become reactivated for various reasons such as cancer, human immunodeficiency virus infection, diabetes, and/or aging. Many studies have examined the mechanisms involved in the latent state of Mycobacterium and showed that latency modified the expression of many genes. Therefore, several mechanisms will change in this bacterium. Hence, this study aimed to briefly examine the genes involved in the latent state as well as the changes that are caused by Mycobacterium tuberculosis. The study also evaluated the relationship between the functions of these genes.

Genomic DNA Extracted from Ancient Antarctic Glacier Ice for Molecular Analyses on the Indigenous Microbial Communities

  • Lee, Sang-Hoon;Bidle, Kay;Falkowski, Paul;Marchant, David
    • Ocean and Polar Research
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    • v.27 no.2
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    • pp.205-214
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    • 2005
  • From ancient Antarctic glacier ice, we extracted total genomic DNA that was suitable for prokaryotic 16S rDNA gene cloning and sequencing, and bacterial artificial chromosome (BAC) library and end-sequencing. The ice samples were from the Dry Valley region. Age dating by $^{40}Ar/^{39}Ar$ analysis on the volcanic ashes deposited in situ indicated the ice samples are minimum 100,000-300,000 yr (sample DLE) and 8 million years (sample EME) old. Further assay proved the ice survived freeze-thaw cycles or other re-working processes. EME, which was from a small lobe of the basal Taylor glacier, is the oldest known ice on Earth. Microorganisms, preserved frozen in glacier ice and isolated from the rest of the world over a geological time scale, can provide valuable data or insight for the diversity, distribution, survival strategy, and evolutionary relationships to the extant relatives. From the 16S gene cloning study, we detected no PCR amplicons with Archaea-specific primers, however we found many phylotypes belonging to Bacteria divisions, such as Actinobacteria, Acidobacteria, Proteobacteria $({\alpha},\;{\beta},\;and\;{\gamma})$, Firmicutes, and Cytophaga-Flavobacterium-Bacteroid$. BAC cloning and sequencing revealed protein codings highly identical to phenylacetic acid degradation protein paaA, chromosome segregation ATPases, or cold shock protein B of present day bacteria. Throughput sequencing of the BAC clones is underway. Viable and culturable cells were recovered from the DLE sample, and characterized by their 16S rDNA sequences. Further investigation on the survivorship and functional genes from the past should help unveil the evolution of life on Earth, or elsewhere, if any.