• 제목/요약/키워드: RNA Sequencing v

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해수 사육 무지개송어(Oncorhynchus mykiss)에서 분리된 Vibrio anguillarum의 특성 분석 (Characteristics of Vibrio anguillarum Isolated from Seawater Cultured Rainbow Trout Oncorhynchus mykiss in Korea)

  • 천혜진;김위식;조미영;정승희;한현자
    • 한국수산과학회지
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    • 제51권3호
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    • pp.254-261
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    • 2018
  • From 2014 to 2017, mortalities of seawater-cultured rainbow trout Oncorhynchus mykiss, were observed in the Goheung and Jeju areas of Korea, with Vibrio anguillarum (seven strains: RT1, 2, 3, 4, 5, 6, and 7) identified as the etiological agent. The phenotypic (based on API 20NE, API ZYM, and E-test kits), serotypic (slide agglutination tests with O1, O2, O3, O4, and O7 antisera), and genotypic (16S rRNA and ompU sequencing) characteristics of the seven RT strains were analyzed and compared to those of seven additional V. anguillarum stains (SF, isolated from sweet fish; FM, isolated from flathead mullet; ATCC43305; ATCC43311; ATCC43307; ATCC43308; and KCTC2711). The phenotypes of the RT strains showed variance, while the slide agglutination tests of the RT1-7, SF, and FM strains all showed positive reactions with serotype O1 antiserum. The 16S rRNA and ompU sequences of the RT1-7, SF, and FM strains were affiliated with V. anguillarum ATCC43305 (Serotype O1), but the ompU sequence of the SF strain differed from those of the RT1-7, FM, and ATCC43311 strains, including one amino acid substitution. We thus confirmed that serotype O1 V. anguillarum, with multiple phenotypes, continues to infect seawater-cultured rainbow trout in Korea.

양식 넙치, Paralichthys olivaceus에서 분리한 Vibrio scophthalmi의 감염 특성 (The Infection Characteristics of Vibrio scophthalmi Isolated from Olive Flounder, Paralichthys olivaceus)

  • 김수현;우승호;이소정;박수일
    • 한국어병학회지
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    • 제26권3호
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    • pp.207-217
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    • 2013
  • 최근 울산광역시 소재의 넙치 양식장에서 체색 흑화, 간 위축, 장관 백탁 등의 증상을 보이며 넙치의 대량 폐사가 빈번히 발생하여, 병원체를 분리하고 감염 특성에 관하여 연구하였다. 2012년 5월 병어로부터 분리한 원인균은 생화학 시험과 16S rRNA, dnaJ gene을 이용한 염기서열 분석을 통해 V. scophthalmi로 동정하였다. 병원성 시험 결과, 본 시험 균주가 $10^6$ CFU/fish에서 75%의 누적 폐사율을 보여 강한 병원성이 확인되었다. V. scophthalmi 감염어는 조직병리학적 병변으로서 간 위축, 장 상피 탈락, 장내 세포 물질 유출 및 장관백탁증 등이 확인되었다.

Potential for the Uptake and Removal of Arsenic [As (V) and As (III)] and the Reduction of As (V) to As (III) by Bacillus licheniformis (DAS1) under Different Stresses

  • Tripti, Kumari;Sayantan, D.;Shardendu, Shardendu;Singh, Durgesh Narain;Tripathi, Anil K.
    • 한국미생물·생명공학회지
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    • 제42권3호
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    • pp.238-248
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    • 2014
  • The metalloid arsenic (Z = 33) is considered to be a significant potential threat to human health due to its ubiquity and toxicity, even in rural regions. In this study a rural region contaminated with arsenic, located at longitude $85^{\circ}$ 32'E and latitude $25^{\circ}$ 11'N, was initially examined. Arsenic tolerant bacteria from the rhizosphere of Amaranthas viridis were found and identified as Bacillus licheniformis through 16S rRNA gene sequencing. The potential for the uptake and removal of arsenic at 3, 6 and 9 mM [As(V)], and 2, 4 and 6 mM [As(III)], and for the reduction of the above concentrations of As(V) to As(III) by the Bacillus licheniformis were then assessed. The minimal inhibitory concentrations (MIC) for As(V) and As(III) was determined to be 10 and 7 mM, respectively. At 3 mM 100% As(V) was uptaken by the bacteria with the liberation of 42% As(III) into the medium, whereas at 6 mM As(V), 76% AS(V) was removed from the media and 56% was reduced to As(III). At 2 mM As(III), the bacteria consumed 100%, whereas at 6 mM, the As(III) consumption was only 40%. The role of pH was significant for the speciation, availability and toxicity of the arsenic, which was measured as the variation in growth, uptake and content of cell protein. Both As(V) and As(III) were most toxic at around a neutral pH, whereas both acidic and basic pH favored growth, but at variable levels. Contrary to many reports, the total cell protein content in the bacteria was enhanced by both As(V) and As(III) stress.

Relative Effect of Glyphosate on Glyphosate-Tolerant Maize Rhizobacterial Communities is Not Altered by Soil Properties

  • Barriuso, Jorge;Mellado, Rafael P.
    • Journal of Microbiology and Biotechnology
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    • 제22권2호
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    • pp.159-165
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    • 2012
  • The rhizobacterial composition varies according to the soil properties. To test if the effect of herbicides on the rhizobacterial communities of genetically modified NK603 glyphosate-tolerant maize varies according to different soil locations, a comparison was made between the effects of glyphosate (Roundup Plus), a post-emergence applied herbicide, and a pre-emergence applied herbicide (GTZ) versus untreated soil. The potential effect was monitored by direct amplification, cloning, and sequencing of the soil DNA encoding 16S rRNA, and high-throughput DNA pyrosequencing of the bacterial DNA coding for the 16S rRNA hypervariable V6 region. The results obtained using three different methods to analyze the herbicide effect on the rhizobacterial communities of genetically modified NK603 maize were comparable to those previously obtained when glyphosate-tolerant maize was grown in soil with different characteristics. Both herbicides decreased the bacterial diversity in the rhizosphere, with Actinobacteria being the taxonomic group most affected. The results suggest that both herbicides affected the structure of the maize rhizobacterial community, but glyphosate was environmentally less aggressive.

Mucin modifies microbial composition and improves metabolic functional potential of a synthetic gut microbial ecosystem

  • Mabwi, Humphrey A.;Komba, Erick V.G.;Mwaikono, Kilaza Samson;Hitayezu, Emmanuel;Mauliasari, Intan Rizki;Jin, Jong Beom;Pan, Cheol-Ho;Cha, Kwang Hyun
    • Journal of Applied Biological Chemistry
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    • 제65권1호
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    • pp.63-74
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    • 2022
  • Microbial dysbiosis in the gut is associated with human diseases, and variations in mucus alter gut microbiota. Therefore, we explored the effects of mucin on the gut microbiota using a community of 19 synthetic gut microbial species. Cultivation of these species in modified Gifu anaerobic medium (GAM) supplemented with mucin before synthetic community assembly facilitated substantial growth of the Bacteroides, Akkermansia, and Clostridium genera. The results of 16S rRNA microbial relative abundance profiling revealed more of the beneficial microbes Collinsella, Bifidobacterium, Ruminococcus, and Lactobacillus. This increased acetate levels in the community cultivated with, rather than without (control), mucin. We identified differences in predicted cell function and metabolism between microbes cultivated in GAM with and without mucin. Mucin not only changed the composition of the gut microbial community, but also modulated metabolic functions, indicating that it could help to modulate microbial changes associated with human diseases.

The fecal microbiota composition of boar Duroc, Yorkshire, Landrace and Hampshire pigs

  • Xiao, Yingping;Li, Kaifeng;Xiang, Yun;Zhou, Weidong;Gui, Guohong;Yang, Hua
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권10호
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    • pp.1456-1463
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    • 2017
  • Objective: To investigate the effect of host genetics on gut microbial diversity, we performed a structural survey of the fecal microbiota of four purebred boar pig lines: Duroc, Landrace, Hampshire, and Yorkshire. Methods: The V3-V4 regions of the 16S rRNA genes were amplified and sequenced. Results: A total of 783 operational taxonomic units were shared by all breeds, whereas others were breed-specific. Firmicutes and Bacteroidetes dominated the majority of the fecal microbiota; Clostridia, Bacilli, and Bacteroidia were the major classes. Nine predominant genera were observed in all breeds and eight of them can produce short-chain fatty acids. Some bacteria can secrete cellulase to aid fiber digestion by the host. Butyric, isobutyric, valeric, and isovaleric acid levels were highest in Landrace pigs, whereas acetic and propionic acid were highest in the Hampshire breed. Heatmap was used to revealed breed-specific bacteria. Principal coordinate analysis of fecal bacteria revealed that the Landrace and Yorkshire breeds had high similarity and were clearly separated from the Duroc and Hampshire breeds. Conclusion: Overall, this study is the first time to compare the fecal microbiomes of four breeds of boar pig by high-throughput sequencing and to use Spearman's rank correlation to analyze competition and cooperation among the core bacteria.

Characterization of the bacterial microbiota across the different intestinal segments of the Qinghai semi-fine wool sheep on the Qinghai-Tibetan Plateau

  • Wang, Xungang;Hu, Linyong;Liu, Hongjin;Xu, Tianwei;Zhao, Na;Zhang, Xiaoling;Geng, Yuanyue;Kang, Shengping;Xu, Shixiao
    • Animal Bioscience
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    • 제34권12호
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    • pp.1921-1929
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    • 2021
  • Objective: The intestinal microbiota enhances nutrient absorption in the host and thus promotes heath. Qinghai semi-fine wool sheep is an important livestock raised in the Qinghai-Tibetan Plateau; however, little is known about the bacterial microbiota of its intestinal tract. The aim of this study was to detect the microbial characterization in the intestinal tract of the Qinghai semi-fine wool sheep. Methods: The bacterial profiles of the six different intestinal segments (duodenum, jejunum, ileum, cecum, colon and rectum) of Qinghai semi-fine wool sheep were studied using 16S rRNA V3-V4 hypervariable amplicon sequencing. Results: A total of 2,623,323 effective sequences were obtained, and 441 OTUs shared all six intestinal segments. The bacterial diversity was significantly different among the different intestinal segments, and the large intestine exhibited higher bacterial diversity than the small intestine. Firmicutes, Bacteroidetes, and Patescibacteria were the dominant phyla in these bacterial communities. Additionally, at the genus level, Prevotella_1, Candidatus_Saccharimonas, and Ruminococcaceae_UCG-005 were the most predominant genus in duodenal segment, jejunal and ileal segments, and cecal, colonic, and rectal segments, respectively. We predicted that the microbial functions and the relative abundance of the genes involved in carbohydrate metabolism were overrepresented in the intestinal segments of Qinghai semi-fine wool sheep. Conclusion: The bacterial communities and functions differed among different intestinal segments. Our study is the first to provide insights into the composition and biological functions of the intestinal microbiota of Qinghai semi-fine wool sheep. Our results also provide useful information for the nutritional regulation and production development in Qinghai semi-fine wool sheep.

Comparison of the oral microbial composition between healthy individuals and periodontitis patients in different oral sampling sites using 16S metagenome profiling

  • Kim, Yeon-Tae;Jeong, Jinuk;Mun, Seyoung;Yun, Kyeongeui;Han, Kyudong;Jeong, Seong-Nyum
    • Journal of Periodontal and Implant Science
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    • 제52권5호
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    • pp.394-410
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    • 2022
  • Purpose: The purpose of this study was to compare the microbial composition of 3 types of oral samples through 16S metagenomic sequencing to determine how to resolve some sampling issues that occur during the collection of sub-gingival plaque samples. Methods: In total, 20 subjects were recruited. In both the healthy and periodontitis groups, samples of saliva and supra-gingival plaque were collected. Additionally, in the periodontitis group, sub-gingival plaque samples were collected from the deepest periodontal pocket. After DNA extraction from each sample, polymerase chain reaction amplification was performed on the V3-V4 hypervariable region on the 16S rRNA gene, followed by metagenomic sequencing and a bioinformatics analysis. Results: When comparing the healthy and periodontitis groups in terms of alpha-diversity, the saliva samples demonstrated much more substantial differences in bacterial diversity than the supra-gingival plaque samples. Moreover, in a comparison between the samples in the case group, the diversity score of the saliva samples was higher than that of the supra-gingival plaque samples, and it was similar to that of the sub-gingival plaque samples. In the beta-diversity analysis, the sub-gingival plaque samples exhibited a clustering pattern similar to that of the periodontitis group. Bacterial relative abundance analysis at the species level indicated lower relative frequencies of bacteria in the healthy group than in the periodontitis group. A statistically significant difference in frequency was observed in the saliva samples for specific pathogenic species (Porphyromonas gingivalis, Treponema denticola, and Prevotella intermedia). The saliva samples exhibited a similar relative richness of bacterial communities to that of sub-gingival plaque samples. Conclusions: In this 16S oral microbiome study, we confirmed that saliva samples had a microbial composition that was more similar to that of sub-gingival plaque samples than to that of supra-gingival plaque samples within the periodontitis group.

Plant Virome Analysis by the Deep Sequencing of Small RNAs of Fritillaria thunbergii var. chekiangensis and the Rapid Identification of Viruses

  • Chen, Lu-xi;Pan, Hang-kai;Tao, Yu-tian;Yang, Dang;Deng, Hui-min;Xu, Kai-jie;Chen, Wen-bin;Li, Jun-min
    • The Plant Pathology Journal
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    • 제38권5호
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    • pp.533-540
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    • 2022
  • Thunberg fritillary (Fritillaria thunbergii), a perennial used in traditional Chinese herbal medicine, is a members of the family Liliaceae. The degeneration of germplasm is a severe problem in the production of Fritillaria thunbergii var. chekiangensis. However, no information about viral infections of F. thunbergii var. chekiangensis has been reported. In this study, we sequenced the small RNAs of F. thunbergii var. chekiangensis from leaves and bulbs, and viruses were identified using a phylogenetic analysis and BLAST search for sequence. In addition, multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) was used to rapidly detect viruses in this variety. Our study first reported that five viruses infected F. thunbergii var. chekiangensis. Among them, fritillary virus Y (FVY), lily mottle virus (LMoV), Thunberg fritillary mosaic virus (TFMV), and hop yellow virus (HYV) had been reported in F. thunbergii, while apple stem grooving virus was first reported in the genus Fritillaria. A multiplex RT-PCR method was developed to rapidly test the four viruses FVY, LMoV, TFMV, and HYV in F. thunbergii var. chekiangensis. Our results provide a better understanding of the infection of F. thunbergii var. chekiangensis by viruses and a basic reference for the better design of suitable control measures.

알코올 내성 젖산균 Pediococcus acidilactici K3와 S1의 분리 및 생리적 특성 (Isolation of the Alcohol-Tolerant Lactic Acid Bacteria Pediococcus acidilactici K3 and S1 and their Physiological Characterization)

  • 장단비;박슬기;이현주;표상은;이한승
    • 한국미생물·생명공학회지
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    • 제41권4호
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    • pp.442-448
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    • 2013
  • 김치, 막걸리 및 시판되는 누룩으로부터 젖산균을 분리한 후 다양한 농도의 알코올을 함유한 MRS 배지에서 배양하여 생육 여부를 조사하였다. 알코올이 13%(v/v) 포함된 MRS 배지에서 $OD_{600}$ 값이 가장 높이 증가한 젖산균 2종을 최종 선별하였고 이 두 균주의 16S rRNA 유전자 염기서열분석 및 API kit를 이용한 생화학적 동정을 실시한 결과 두 종 모두 P. acidilactici에 가까운 종인 것을 확인하여 P. acidilactici K3와 P. acidilactici S1으로 명명하였다. 선별한 P. acidilactici K3와 S1의 bacteriocin 생성 여부를 확인한 결과 식중독균인 S. aureus의 생육 저해를 일으키는 bacteriocin 생성 능력이 있음을 확인하였다. 선별한 두 균주와 표준 균주(P. acidilactici DSM 20284)의 알코올 내성을 비교한 결과 6% 이하의 저농도 알코올이 포함된 배지에서는 균주 생육에 큰 차이가 없었으나 12% 이상의 알코올이 포함된 배지에서는 표준 균주보다 최대 3배 이상 알코올 내성이 뛰어남을 확인하였다. 알코올 내성 균주 P. acidilactici K3와 S1을 살균 막걸리에 첨가한 후 냉장 온도($4^{\circ}C$$10^{\circ}C$)에서 저장하며 생균수를 측정한 결과 사멸하지 않고 한 달 간 생균수가 유지됨을 확인하였다.