• 제목/요약/키워드: 16S rRNA sequencing

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Microbial Contamination according to the Numbers of Mask Worn in the Community

  • Eun Ju Lee;Heechul Park;Min-A Je;Songhee Jung;Gahee Myoung;Su Bin Jo;Hyun Min Hwang;Ryeong Si;Hyunwoo Jin;Kyung-Eun Lee;Jungho Kim
    • 대한의생명과학회지
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    • 제28권4호
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    • pp.317-321
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    • 2022
  • Due to COVID-19 pandemic, wearing face masks is obligatory to prevent respiratory virus transmissions in the community. However, there are few studies of the desirable number of wearing a face mask, and how to store them for reuse. Therefore, in this study, a survey was conducted among 208 healthy adults, and 27 kf-94 masks worn for 1, 2, and 3 days were collected. To estimate the risk of bacterial contamination, we analyzed the extent of bacterial contamination of the BHI medium and 16S rRNA gene sequencing. With an increase in the number of days of using the mask, the degree of bacterial contamination of the used mask gradually increased. As a result of 16S rRNA PCR performed for strain identification, Staphylococcus, known as a pathogenic bacterium, was identified the most. In conclusion, we found that wearing a cotton KF mask provides an optimal environment for microbes, which are related to the skin and respiratory system, to thrive. Therefore, it is also important to reduce the risk of bacterial infection of the face mask with appropriate sterilization methods.

Identification of plasma miRNA biomarkers for pregnancy detection in dairy cattle

  • Lim, Hyun-Joo;Kim, Hyun Jong;Lee, Ji Hwan;Lim, Dong Hyun;Son, Jun Kyu;Kim, Eun-Tae;Jang, Gulwon;Kim, Dong-Hyeon
    • 한국동물생명공학회지
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    • 제36권1호
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    • pp.35-44
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    • 2021
  • A pregnancy diagnosis is an important standard for control of livestock's reproduction in paricular dairy cattle. High reproductive performance in dairy animals is a essential condition to realize of high life-time production. Pregnancy diagnosis is crucial to shortening the calving interval by enabling the farmer to identify open animals so as to treat or re-breed them at the earliest opportunity. MicroRNAs are short RNA molecules which are critically involved in regulating gene expression during both health and disease. This study is sought to establish the feasible of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from 12 non-pregnant cows ("open" cows: samples were collected before insemination (non-pregnant state) and after pregnancy check at the indicated time points) on weeks 0, 4, 8, 12 and 16. Using small RNA sequencing we identified a total of 115 miRNAs that were differentially expressed weeks 16 relative to non-pregnancy ("open" cows). Weeks 8, 12 and 16 of pregnancy commonly showed a distinct increase in circulating levels of miR-221 and miR-320a. Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with pregnancy in cattle. Their application in the field of reproductive biology has opened up opportunities for research communities to look for pregnancy biomarker molecules in dairy cattle.

16S rRNA 염기서열 분석을 통한 오분자기(Sulculus diversicolor supertexta)내 미생물 군집 조사 및 인체유해 질병세균에 대한 항균활성 모니터링 (Investigation of Microbial Communities in Sulculus diversicolor supertexta Through 16S rRNA Sequencing and Antibacterial Monitoring of Harmful Strains)

  • 김민선;이승종;허문수
    • 생명과학회지
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    • 제28권12호
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    • pp.1477-1488
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    • 2018
  • 본 연구는 제주 연안에서 채집한 오분자기(Sulculus diversicolor supertexta)를 구성하는 미생물 군집의 다양성을 알아보기 위하여 근육, 장, 생식소 각 부위별로 조사하였다. 배지로 1차 순수 분리한 결과 근육은 MA, 장 1% BHIA, 생식소 1% TSA에서 각각 최대 군락 계수가 나타났다. 16S rRNA sequence로 표준 균주와 비교 유사도 분석 결과 총 190개의 순수 colony가 분리되었다. NBLAST program 분석 결과 크게 5문 25과 39속 71종으로 나타났다. 표준 균주와 상동성은 91-100%를 나타냈다. 오분자기 내 미생물 군집은 크게 Probacteria (Gamma-proteobacteria, Alpha-proteobacteria) 48%, Actinobacteria 32.5%, Firmicutes 16.9%, Bacteroide 1.3%, Deinococcus-thermus 1.3%로 나타났다. 근육, 장, 생식소 모든 부위에서 Moraxellaceae과 Psychrobacter cibarius가 우점하였다. 근육, 장, 생식소 모든 부위에서 Alteromonadaceae, Enterobacteriaceae, Pasturellaceae, Moraxellaceae, Rhodobacteraceae, Geminicoccaceae, Dietziaceae, Intrasporangiaceae, Microbacteriaceae, Micrococcaceae, Micromonosporaceae, Streptomycetaceae, Aerococcaceae, Bacillaceae, Paenibacillaceae, Planococcaceae, Staphylcoccaceae가 공통적으로 분리되었으며, 장에서 Flavobacteriaceae, Corynebacteriaceae, Yesiniaceae, Vibrionaceae, Hahellaceae, Pseudomonadaceae가 추가 분리되었다. 분리 균주로부터 인체 유해 질병 세균에 대한 항균활성 모니터링 결과 Sterptomyces albus (96%)가 4균주 모두 항균활성을 보였고 Agrococcus baldri (99%), Psychrobacter nivimaris (99%)가 E. coli, E. aerogens에 대한 항균활성을 나타냈다. 그 외 상동성이 낮은 일부 균주가 분리되어 신균주 실험을 비롯한 항균활성물질 정제 등 추가 실험이 필요한 것으로 사료된다. 본 실험은 오분자기 미생물 군집의 다양성과 유전학적 자원을 확보하는데 의의를 두며, 응용 미생물의 개발 가능성에 있어 기초 자료를 제공하고자 하였다.

16S rRNA 유전자 계통분석에 의한 한강수계의 세균 다양성 (Bacterial Diversity of the Han River as Determined by 16S rRNA Gene Analysis)

  • 한석균;이일규;안태영
    • 미생물학회지
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    • 제34권4호
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    • pp.194-199
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    • 1998
  • 한강의 본류와 만나는 탄천과 중랑천에서 16S rDAN를 증폭하고 부분적인 염기서열 분석을 통하여 한강의 세균 다양성을 결정하였다. 총 27개의 클론을 분리하였으며 RFLP를 이용하여 7개의 group으로 나누었다. 탄천의 15개 클론은 4개의 group으로 나뉘어졌으며 가장 많은 클론을 포함하는 group(HT-1 클론)은 class Proteobacteria의 ${\delta}$-subdivision에 속하는 Acrobacter cryaerophilius와 높은 유사도를 보였으며, 다른 두 group(HT-6과 HT-9 클론)은 모두 clas Cytophagales에 속하였다. 중랑천의 12개의 클론은 3개의 group으로 나뉘어졌으며 가장 많은 클론을 보이는 group(HJ-1 클론)은 class Proteobacteria의 ${\alpha}$-subdivision에 속하는 Sphingomonas sp. 와 높은 유사도를 나타내었다. 전체적으로는 Proteobateria(alpha, beta and delta subdivision), Cytophagales와 Actinomycetales가 검출되었다.

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Bioinformatic Suggestions on MiSeq-Based Microbial Community Analysis

  • Unno, Tatsuya
    • Journal of Microbiology and Biotechnology
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    • 제25권6호
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    • pp.765-770
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    • 2015
  • Recent sequencing technology development has revolutionized fields of microbial ecology. MiSeq-based microbial community analysis allows us to sequence more than a few hundred samples at a time, which is far more cost-effective than pyrosequencing. The approach, however, has not been preferably used owing to computational difficulties of processing huge amounts of data as well as known Illumina-derived artefact problems with amplicon sequencing. The choice of assembly software to take advantage of paired-end sequencing and methods to remove Illumina artefacts sequences are discussed. The protocol we suggest not only removed erroneous reads, but also dramatically reduced computational workload, which allows even a typical desktop computer to process a huge amount of sequence data generated with Illumina sequencers. We also developed a Web interface (http://biotech.jejunu.ac.kr/ ~abl/16s/) that allows users to conduct fastq-merging and mothur batch creation. The study presented here should provide technical advantages and supports in applying MiSeq-based microbial community analysis.

Practical considerations for the study of the oral microbiome

  • Yu, Yeuni;Lee, Seo-young;Na, Hee Sam
    • International Journal of Oral Biology
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    • 제45권3호
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    • pp.77-83
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    • 2020
  • In the oral cavity, complex microbial community is shaped by various host and environmental factors. Extensive literature describing the oral microbiome in the context of oral health and disease is available. Advances in DNA sequencing technologies and data analysis have drastically improved the analysis of the oral microbiome. For microbiome study, bacterial 16S ribosomal RNA gene amplification and sequencing is often employed owing to the cost-effective and fast nature of the method. In this review, practical considerations for performing a microbiome study, including experimental design, molecular analysis technology, and general data analysis, will be discussed.

생물학적 인 제거용 연속회분식 반응기에서의 미생물 분포 조사

  • 전체옥;박종문
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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    • pp.77-80
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    • 2000
  • Various analytical methods such as electron microscopy, quinone analysis, and 16S rDNA sequencing studies were used to investigate the microbial communities and to identify the microorganisms responsible for enhanced biological phosphorus removal (EBPR) in an anaerobic/aerobic sequencing batch reactor (SBR) fed with acetate. Electron photomicrographs showed that oval-shaped microorganisms of about $0.7\;{\sim}\;1\;{\mu}m$ in diameter dominated the microbial sludge. These microorganisms contained polyphosphate granules and glycogen inclusions, which suggests that they are a kind of phosphorus accumulating organism. Quinone and 16S rRNA sequence analyses showed that the members of Proteobacteria beta subclass were the most abundant species, which were affiliated with the Rhodocyclus-likes group. Phylogenetic analysis revealed that the two dominating clones of the beta subclass were most distantly related to Propionivibrio dicarboxylicus DSM 5885 and Rhodocyclus tenuis DSM 109 with about 95% and 96% sequence similarity, respectively. Therefore, it was concluded that the oval-shaped organisms related to the Rhodocyclus-likes group are likely to be responsible for biological phosphorus removal in SBR operation supplied with acetate.

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Microbiological Characteristics of Nocardia takedensis Isolated from Skin Lesion, in Korea

  • Kang, Hye-Sook;Park, Gyu-Nam;Kim, Hye-Ran;Chang, Kyung-Soo
    • 대한의생명과학회지
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    • 제23권2호
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    • pp.96-103
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    • 2017
  • Nocardia species (spp.) are opportunistic pathogen in immunocompromised hosts. The genus Nocardia contains more than 70 species. Nocardia takedensis has been recently reported as a new species of the genus Nocardia. In this study, we describes the first clinical isolate of N. takedensis from the skin lesion in Busan, Korea. For the identification of clinical isolate to the species level as N. takedensis, classical methods (colony morphology, biochemical characteristics, and antimicrobial susceptibility), molecular method (16S rRNA gene sequencing), and MS (mass spectrometry) analysis were conducted. Clinical isolates grew slowly on the culture media (5% sheep blood agar and chocolate agar) under 5% $CO_2$ condition. Especially, carotene pigmentation was detected well on the media. Using mass spectrometry, Nocardia isolate was not identified to the species level. However, molecular method based on 16S rRNA sequencing confirmed the isolate as N. takedensis correctly. N. takedensis isolate was partial positive for acid-fast bacilli on the Ziehl-Neelsen method. And it was observed to be resistance to amoxicillin/clavulanic acid and ciprofloxacin. Our results provide useful information to develop optimal identification protocol of N. takedensis in clinical diagnostic laboratories.

Development of a Rapid and Accurate Identification Method for Citrobacter Species Isolated from Pork Products Using a Matrix-Assisted Laser-Desorption Ionization Time-of-Flight Mass Spectrometry (MALDITOF MS)

  • Kwak, Hye-Lim;Han, Sun-Kyung;Park, Sunghoon;Park, Si Hong;Shim, Jae-Yong;Oh, Mihwa;Ricke, Steven C.;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • 제25권9호
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    • pp.1537-1541
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    • 2015
  • Previous detection methods for Citrobacter are considered time consuming and laborious. In this study, we have developed a rapid and accurate detection method for Citrobacter species in pork products, using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). A total of 35 Citrobacter strains were isolated from 30 pork products and identified by both MALDI-TOF MS and 16S rRNA gene sequencing approaches. All isolates were identified to the species level by the MALDI-TOF MS, while 16S rRNA gene sequencing results could not discriminate them clearly. These results confirmed that MALDITOF MS is a more accurate and rapid detection method for the identification of Citrobacter species.

Comparative Analysis of Gut Microbial Communities in Children under 5 Years Old with Diarrhea

  • Wen, Hongyu;Yin, Xin;Yuan, Zhenya;Wang, Xiuying;Su, Siting
    • Journal of Microbiology and Biotechnology
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    • 제28권4호
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    • pp.652-662
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    • 2018
  • Diarrhea is a global disease with a high morbidity and mortality rate in children. In this study, 25 fecal samples were collected from children under 5 years old. Seven samples had been taken from healthy children without diarrhea and marked as the healthy control group; eight samples had been sampled from children with diarrhea caused by dyspepsia and defined as the non-infectious group; and ten samples had been taken from children with diarrhea induced by intestinal infections and identified as the infectious group. We detected the microbial communities of samples by using high-throughput sequencing of 16S rRNA genes. The proportion of aerobic and facultative anaerobic microbes in samples of the infectious group was much higher than in the non-infectious group. In addition, the relative abundance of Enterococcus in the healthy control group was significantly higher than in the non-infectious group and infectious group. This can be used as a potential diagnostic biomarker for diarrhea.