• Title/Summary/Keyword: 차세대 서열 분석

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Comparison of gut microbial diversity of breast-fed and formula-fed infants (모유수유와 분유수유에 따른 영아 장내 미생물 군집의 특징)

  • Kim, Kyeong Soon;Shin, Jung;Sim, JiSoo;Yeon, SuJi;Lee, Pyeong An;Chung, Moon Gyu
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.268-273
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    • 2019
  • The intestinal microbiomes vary according to the factors such environment, age and diet. The purpose of this study was to compare the gut microbial diversity between Korean infants receiving breast-fed milk and formula-fed milk. We analyzed microbial communities in stool samples collected from 80 Korean infants using next generation sequencing. Phylum level analysis revealed that microbial communities in both breast-fed infants group (BIG) was dominated by Actinobacteria ($74.22{\pm}3.48%$). Interestingly, the phylum Actinobacteria was dominant in formula-fed infants group A (FIG-A) at $73.46{\pm}4.12%$, but the proportions of phylum Actinobacteria were lower in formulafed infants group B and C (FIG-B and FIG-C) at $66.52{\pm}5.80%$ and $68.88{\pm}4.33%$. The most abundant genus in the BIG, FIG-A, FIG-B, and FIG-C was Bifidobacterium, comprising $73.09{\pm}2.31%$, $72.25{\pm}4.93%$, $63.81{\pm}6.05%$, and $67.42{\pm}5.36%$ of the total bacteria. Furthermore, the dominant bifidobacterial species detected in BIG and FIG-A was Bifidobacterium longum at $68.77{\pm}6.07%$ and $66.85{\pm}4.99%$ of the total bacteria. In contrast, the proportions of B. longum of FIG-B and FIG-C were $58.94{\pm}6.20%$ and $61.86{\pm}5.31%$ of the total bacteria. FIG-A showed a community similar to BIG, which may be due to the inclusion of galactooligosaccharide, galactosyllactose, synergy-oligosaccharide, bifidooligo and improvement material of gut microbiota contained in formula-milk. We conclude that 5-Bifidus factor contained in milk powder promotes the growth of Bifidobacterium genus in the intestines.

Efficiency of Density Gradient Centrifugation Method (Ludox method) Based on eDNA for the Analysis of Harmful Algal Bloom Potential (유해남조류 발생 잠재성 분석을 위한 eDNA 기반의 퇴적물 전처리 방법: 밀도 구배 원심분리법(Ludox method))

  • Kyeong-Eun Yoo;Hye-In Ho;Hyunjin Kim;Keonhee Kim;Soon-Jin Hwang
    • Korean Journal of Ecology and Environment
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    • v.56 no.1
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    • pp.36-44
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    • 2023
  • Environmental DNA (eDNA) can exist in both intracellular and extracellular forms in natural ecosystems. When targeting harmful cyanobacteria, extracellular eDNA indicates the presence of traces of cyanobacteria, while intracellular eDNA indicates the potential for cyanobacteria to occur. However, identifying the "actual" potential for harmful cyanobacteria to occur is difficult using the existing sediment eDNA analysis method, which uses silica beads and cannot distinguish between these two forms of eDNA. This study analyzes the applicability of a density gradient centrifugation method (Ludox method) that can selectively analyze intracellular eDNA in sediment to overcome the limitations of conventional sediment eDNA analysis. PCR was used to amplify the extracted eDNA based on the two different methods, and the relative amount of gene amplification was compared using electrophoresis and Image J application. While the conventional bead beating method uses sediment as it is to extract eDNA, it is unknown whether the mic gene amplified from eDNA exists in the cyanobacterial cell or only outside of the cell. However, since the Ludox method concentrates the intracellular eDNA of the sediment through filtration and density gradient, only the mic gene present in the cyanobacteria cells could be amplified. Furthermore, the bead beating method can analyze up to 1 g of sediment at a time, whereas the Ludox method can analyze 5 g to 30 g at a time. This gram of sediments makes it possible to search for even a small amount of mic gene that cannot be searched by conventional bead beating method. In this study, the Ludox method secured sufficient intracellular gene concentration and clearly distinguished intracellular and extracellular eDNA, enabling more accurate and detailed potential analysis. By using the Ludox method for environmental RNA expression and next-generation sequencing (NGS) of harmful cyanobacteria in the sediment, it will be possible to analyze the potential more realistically.

Cross Resistance of Fluoroquinolone Drugs on gyrA Gene Mutation in Mycobacterium tuberculosis (결핵균에서 gyrA 유전자 돌연변이에 따른 Fluoroquinolone계 약제들의 교차내성)

  • Park, Young Kil;Park, Chan Hong;Koh, Won-Jung;Kwon, O Jung;Kim, Bum Jun;Kook, Yoon Hoh;Cho, Sang Nae;Chang, Chulhun;Bai, Gill Han
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.3
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    • pp.250-256
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    • 2005
  • Background : Fluoroquinolone drugs are an important anti-tuberculous agent for the treatment of multi-drug resistant tuberculosis. However, many drugs belonging to the fluoroquinolones have different cross resistance to each other. Methods : Sixty-three ofloxacin (OFX) resistant and 10 pan-susceptible M. tuberculosis isolates were selected, and compared for their cross resistance using a proportion method on Lowenstein-Jensen media, containing ofloxacin (OFX), ciprofloxacin (CIP), levofloxacin (LVX), moxifloxacin (MXF), gatifloxacin (GAT) and sparfloxacin (SPX), at concentrations ranging from 0.5 to $3{\mu}g/ml$. DNA extracted from the isolates was directly sequenced after amplifying from the gyrA and gyrB genes. Results : The 63 OFX resistant M. tuberculosis isolates showed complete cross resistance to CIP, but only 90.5, 44.4, 36.5 and 46.0% to LVX, MXF, GAT, and to SPX, respectively. Fifty-one of the isolates (81.0%) had point mutations in codons 88, 90, 91 and 94 in gyrA, which are known to be correlated with OFX resistance. The Gly88Ala, Ala90Valand Asp94Ala mutations in gyrA showed a tendency to be susceptible to MXF, GAT and SPX. Only 4 isolates had mutations in the gyrB gene, which did not affect the OFX resistance. Conclusion : About 60% of the OFX resistant M. tuberculosis isolates were susceptible to GAT, SPX and MXF. These fluoroquinolones may be useful in the treatment of TB patients showing OFX resistance.

Feasibility of Environmental DNA Metabarcoding for Invasive Species Detection According to Taxa (분류군별 외래생물 탐지를 위한 환경 DNA 메타바코딩 활용 가능성)

  • Yujin Kang;Jeongeun Jeon;Seungwoo Han;Suyeon Won;Youngkeun Song
    • Journal of Environmental Impact Assessment
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    • v.32 no.2
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    • pp.94-111
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    • 2023
  • In order to establish an effective management strategy for invasive species early detection and regular monitoring are required to assess their introduction or dispersal. Environmental DNA (eDNA) is actively applied to evaluate the fauna including the presence of invasive species as it has high detection sensitivity and can detect multiple species simultaneously. In Korea, the applicability evaluation of metabarcoding is being conducted mainly on fish, and research on other taxa is insufficient. Therefore, this study identified the feasibility of detecting invasive species in Korea using eDNA metabarcoding. In addition, to confirm the possibility of detection by taxa, the detection of target species was evaluated using four universal primers (MiFish, MiMammal, Mibird, Amp16S) designed for fish, mammals, birds, and amphibians. As a result, target species (Trachemys scripta, 3 sites; Cervus nippon, 3 sites; Micropterus salmoides, 7 sites; Rana catesbeiana, 4 sites) were detected in 17 of the total 55 sites. Even in the selection of dense sampling sites within the study area, there was a difference in the detection result by reflecting the ecological characteristics of the target species. A comparison of community structures (species richness, abundance and diversity) based on the presence of invasive species focused on M.salmoides and T.scripta, showed higher diversity at the point where invasive species were detected. Also, 1 to 4 more species were detected and abundance was also up to 1.7 times higher. The results of invasive species detection through metabarcoding and the comparison of community structures indicate that the accumulation of large amounts of monitoring data through eDNA can be efficiently utilized for multidimensional ecosystem evaluation. In addition, it suggested that eDNA can be used as major data for evaluation and prediction, such as tracking biological changes caused by artificial and natural factors and environmental impact assessment.

Physiological Characteristics of Melon Plants Showing Leaf Yellowing Symptoms Caused by CABYV Infection (CABYV 감염 멜론의 황화증상에 따른 생리적인 특성)

  • Lee, Hee Ju;Kim, Mi-Kyeong;Lee, Sang Gyu;Choi, Chang Sun;Choi, Hong-Soo;Kwak, Hae Ryun;Choi, Gug Seoun;Chun, Changhoo
    • Horticultural Science & Technology
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    • v.33 no.2
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    • pp.210-218
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    • 2015
  • Melon leaves showing yellowing symptoms were analyzed using electron microscopy and RT-PCR for major cucurbit-infecting-viruses (CMV, MNSV, CGMMV, SqMV, WMV, KGMMV, PRSV and ZYMV) reported in Korea, but these viruses were not detected. As the result of further analysis by next-generation sequencing (NGS), the virus was identified as Cucurbit aphid-borne yellows virus (CABYV), and then confirmed by RT-PCR using CABYV-specific primers. When photosynthetic capacity was measured based on chlorophyll fluorescence yield (ChlFY), the leaves of the diseased plants showed $4.09{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, which was one-third of the readings observed for unaffected normal plants ($12.36{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$). The root functions of plants affected by leaf yellowing symptoms (LYS) was $0.28mg{\cdot}g^{-1}$, about half that measured for the normal unaffected plants ($0.48mg{\cdot}g^{-1}$). Cytological observations revealed that there were no morphological differences in the palisade parenchyma and mesophyll spongy cells of the leaves between the diseased and the normal plants. However, the same leaf cells of the affected plants contained more starch granules compared to those of the normal, unaffected plants. We conclude that the LYS of muskmelon is not merely a physiological disorder but a viral disease caused by CABYV and spread by aphids.