• Title/Summary/Keyword: 파이로시퀀싱

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Analysis of gut bacterial diversity and exploration of cellulose-degrading bacteria in xylophagous insects (목질섭식곤충의 장내 세균 다양성 분석 및 섬유소 분해균 탐색)

  • Choi, Min-Young;Ahn, Jae-Hyung;Song, Jaekyeong;Kim, Seong-Hyun;Bae, Jin-Woo;Weon, Hang-Yeon
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.209-220
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    • 2015
  • In this study, gut bacterial communities in xylophagous insects were analyzed using the pyrosequencing of 16S rRNA genes for their potential biotechnological applications in lignocelluloses degradation. The result showed that operational taxonomic units (OTUs), species richness and diversity index were higher in the hindgut than in the midgut of all insect samples analyzed. The dominant phyla or classes were Firmicutes (54.0%), Bacteroidetes (14.5%), ${\gamma}-Proteobacteria$ (12.3%) in all xylophagous insects except for Rhinotermitidae. The principal coordinates analysis (PCoA) showed that the bacterial community structure mostly clustered according to phylogeny of hosts rather than their habitats. In our study, the two carboxymethyl cellulose (CMC)-degrading isolates which showed the highest enzyme activity were most closely related to Bacillus toyonensis $BCT-7112^T$ and Lactococcus lactis subsp. hordniae $NCDO\;2181^T$, respectively. Cellulolytic enzyme activity analysis showed that ${\beta}-1,4-glucosidase$, ${\beta}-1,4-endoglucanase$ and ${\beta}-1,4-xylanase$ were higher in the hindgut of Cerambycidae. The results demonstrate that xylophagous insect guts harbor diverse gut bacteria, including valuable cellulolytic bacteria, which could be used for various biotechnological applications.

Analysis of Microbial Communities in Animal Carcass Disposal Soils (가축사체 매몰지 토양의 미생물 군집 분석)

  • Park, Jeong-Ann;Choi, Nag-Choul;Kim, Song-Bae
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.7
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    • pp.503-508
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    • 2013
  • The aim of this study was to investigate the microbial communities in animal carcass disposal soils to examine the possible threat of pathogens from leachate. DNA extraction was performed for the soils in three carcass disposal sites located in Gyeonggi-do, Korea, and then 16S rRNA pyrosequencing was conducted to identify the microbial communities. Results indicate that, according to phylum classification, Proteobacteria (100%) was identified in soil A, Actinobacteria (66.4%) > Proteobacteria (31.1%) > Bacteriodetes (2.1%) > Acidobacteria (0.3%) in soil B, and Actinobacteria (63.1%) > Proteobacteria (36.9%) in soil C. According to genus classification, Pseudomonas was dominant in soil A (98%), Arthrobacter in soil B (68%) and C (61%). There were no detections of pathogens such as Salmonella, Campylobacter and Clostridium perfringens. However, high concentration of Ralstonia pickettii causing bacteremia was observed. Although carcass disposal soils examined in this study were not highly contaminated with pathogens, further monitoring is still needed to examine the potential threat of pathogens in leachate derived from carcass disposal sites.

Bacterial core community in soybean rhizosphere (콩 근권의 핵심 세균 군집)

  • Lee, Youngmi;Ahn, Jae-Hyung;Choi, Yu-Mi;Weon, Hang-Yeon;Yoon, Jung-Hoon;Song, Jaekyeong
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.347-354
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    • 2015
  • Soybean is well known to be originated from Korea and far-east Asian countries, and studies of many root nodule bacteria associated with soybean have mainly-focused on nitrogen fixation, but much less study was carried out on bacterial community in the rhizosphere of soybean. In this study, we analyzed the bacterial community in rhizosphere of Korean soybean, Daepungkong using the pyrosequencing method based on the 16S rRNA gene to characterize the change of the rhizosphere community structure according to the growth stages of soybeans and to elucidate bacterial core community in rhizosphere of soybean. Our results revealed that bacterial community of rhizosphere soil differed from that of bulk soil and was composed of a total of 21 bacterial phyla. The predominant phylum in the rhizosphere of soybean was Proteobacteria (36.6-42.5%) and followed by Acidobacteria (8.6-9.4%), Bacteroidetes (6.1-10.9%), Actinobacteria (6.4-9.8%), and Firmicutes (5.7-6.3%). The bacterial core community in soybean rhizosphere was mainly composed of the operational taxonomic units (OTUs) belonging to the phylum Proteobacteria throughout all growth stages. The OTU00006 belonged to the genus Bradyrhizobium had the highest abundance and Steroidobacter, Streptomyces, Devosia were followed. These results show that bacterial core community in soybean rhizosphere was mainly composed of OTUs associated with plant growth promotion and nutrient cycles.

Comparative Analysis of Soil Microbial Communities between Conventional and Organic Farming Systems in Pepper Cultivation (관행과 유기농 고추 재배지의 토양미생물 군집 비교)

  • Kim, Yiseul;Lee, Youngmi;Weon, Hang-Yeon;Sang, Mee Kyung;Song, Jaekyeong
    • Korean Journal of Organic Agriculture
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    • v.28 no.2
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    • pp.235-250
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    • 2020
  • Agricultural practices are known to have a crucial influence not only on soil physico-chemical properties but also on microbial communities. To investigate the effect of farming practices on soil microbial communities, a total of 10 soil samples were collected, including five conventional and five organic farming soils cultivated with peppers in plastic greenhouse. We conducted barcorded-pyrosequencing of V1-V3 regions of 16S rRNA genes to examine soil microbial communities of two different farming practices. Taxonomic classification of the microbial communities at the phylum level indicated that a total of 22 bacterial phyla were present across all samples. Among them, seven abundant phyla (>3%) including Proteobacteria, Actinobacteria, Firmicutes, Acidobacteria, Bacteroidetes, Chloroflexi, and Gemmatimonadetes were found, and Proteobacteria (33.0 ± 5.7%), Actinobacteria (19.9 ± 9.7%), and Firmicutes (13.6 ± 5.0%) comprised more than 66% of the relative abundance of the microbial communities. Organic farming soils showed higher relative abundances of Proteobacteria and Firmicutes, while Actinobacteria and Chloroflexi were more abundant in conventional farming soils. Notably, the genera Bacillus (higher in organic farming soils) and Streptomyces (higher in conventional farming soils) exhibited significant variation in relative abundance between organic and conventional farming soils. Finally, correlation analysis identified significant relationships (p<0.05) between soil chemical properties, in particular, pH and organic matter content and microbial communities. Taken together, this study demonstrated that the changes of soil physico-chemical properties by agricultural farming practices effected significantly (p<0.05) on soil microbial communities.

Identification of microbiome with 16S rRNA gene pyrosequencing and antimicrobial effect of egg white in bovine mastitis (젖소 유방염에서 16S rRNA 파이로시퀀싱을 이용한 우유 내 마이크로바이옴의 동정과 난백의 항균효과)

  • Kim, Danil;Kim, Eun-Kyung;Seong, Won-Jin;Ro, Younghye;Ko, Dae-Sung;Kim, Nam-Hyung;Kim, Jae-Hong;Kwon, Hyuk-Joon
    • Korean Journal of Veterinary Research
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    • v.57 no.2
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    • pp.117-126
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    • 2017
  • Bovine mastitis is an important microbial disease in the dairy industry. We investigated the frequencies of bacterial pathogens in 62 farms and pathogen antibiotic resistance from mastitis samples (n = 748). We tested the antimicrobial activity of chicken and duck egg white and lysozyme purified from chicken egg white. Moreover, we compared the microbiomes of normal and mastitic raw milk obtained by 16S rRNA gene pyrosequencing and culture methods. The results showed that the frequencies of Gram-positive pathogens (Enterococcus faecalis 37% and Staphylococcus aureus 36%) were higher than that of a Gram-negative pathogen (Escherichia coli 15%). Resistance frequencies to ampicillin and norfloxacin were lowest in Staphylococcus aureus (21%), Enterococcus faecalis (23%), and Escherichia coli (33%), and the antimicrobial activity of chicken egg white was higher than those of lysozyme and duck egg white. Pyrosequencing results revealed clear differences between the microbiomes of mastitic and normal raw milk samples and revealed a slightly similar, but clearly different, composition of pathogens compared to that from the culture method. Thus, pyrosequencing may be useful for elucidating changes in microbiomes during mastitis progression and treatment. A chicken egg white and antibiotic combination may help with mastitis treatment; however, further studies are needed.

EGFR Analysis in Cytologic Samples of Lung Adenocarcinoma by Microdissection (미세 절제에 의한 폐 선암 세포 검체에서 EGFR 분석)

  • Han, Jeong Yeon;Lee, Hoon Taek;Oh, Seo Young
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.3
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    • pp.125-131
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    • 2015
  • The discovery of activating mutations in EGFR in a subset of lung adenocarcinomas was a major advance in our understanding of lung adenocarcinoma biology, and has led to groundbreaking studies that have demonstrated the efficacy of tyrosine kinase inhibitor therapy. Cytologic specimen procedures have become increasingly popular for obtaining diagnostic material in lung carcinomas. However, frequently the small amount of material or sparseness of tumor cells obtained from cytologic preparations limit the number of specialized studies, such as mutation analysis, that can be performed. In this study we used microdissection to isolate small numbers of tumor cells to assess for EGFR mutations from 76 cytological smear slides of patients with lung adenocarcinomas. We compared our results with previous molecular assays that had been performed on either surgical or cytology specimens as part of the patient's initial clinical work-up. Not only were we able to detect the identical EGFR mutation through the pyrosequencing, but we were also able to consistently detect the mutation from as few as 25 microdissected tumor cells. Furthermore, isolating a purer population of tumor cells resulted in increased sensitivity of mutation detection as we were able to detect mutations from microdissection-enriched cases. Therefore, microdissection can not only significantly increase the number of lung adenocarcinoma patients that can be screened for EGFR mutations, but can also facilitate the use of cytologic samples in the newly emerging field of molecular-based personalized therapies.

Analysis of Rhizosphere Soil Bacterial Communities on Seonginbong, Ulleungdo Island (울릉도 성인봉의 근권 토양 세균군집 분석)

  • Nam, Yoon-Jong;Yoon, Hyeokjun;Kim, Hyun;Kim, Jong-Guk
    • Journal of Life Science
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    • v.25 no.3
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    • pp.323-328
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    • 2015
  • The study of microbial diversity and richness in soil samples from a volcanic island named Ulleungdo, located east of South Korea. The soil bacterial communities on the Ulleungdo were analyzed using pyrosequencing method based on 16S rRNA gene. There were 1,613 operational taxonomic units (OUT) form soil sample. From results of a BLASTN search against the EzTaxon-e database, the validated reads (obtained after sequence preprocessing) were almost all classified at the phylum level. Proteobacteria was the most dominant phylum with 48.28%, followed by acidobacteria (26.30%), actionbacteria (6.89%), Chloroflexi (4.58), Planctomycetes (4.56%), Nitrospirae (1.83%), Bacteroidetes (1.51%), Verrucomicrobia (1.48%), and Gemmatimonadetes (1.11%). α-proteobacteria was the most dominant class with 36.07% followed by Acidobacteria_c (10.65%), Solibacteres (10.64%), δ-proteobacteria (4.42%), γ-proteobacteria (4.29%), Planctomycetacia (4.16%), Actinobacteria_c (4.00%), Betaproteobacteria (3.50%), EU686603_c (2.97%), Ktedonobacteria (2.91%), Acidimicrobiia (1.32%), Verrucomicrobiae (1.27%), Gemmatimonadetes_c (1.11%), Sphingobacteria (1.09%), and GU444092_c (1.06%). Bradyrhizobiaceae was the most dominant family with 22.83% followed by Acidobacteriaceae (10.62%), EU445199_f (5.72%), Planctomycetaceae (4.03%), Solibacteraceae (3.63%), FM209092_f (3.58%), Steroidobacter_f (2.81%), EU686603_f (2.73%), Hyphomicrobiaceae (2.33%), Ktedonobacteraceae (1.75%), AF498716_f (1.46%), Rhizomicrobium_f (1.03%), and Mycobacteriaceae (1.01%). Differences in the diversity of bacterial communities have more to do with geography than the impact on environmental factors and also the type of vegetation seems to affect the diversity of bacterial communities.