• Title/Summary/Keyword: MiSeq

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Microbial Community of Tannery Wastewater Involved in Nitrification Revealed by Illumina MiSeq Sequencing

  • Ma, Xiaojian;Wu, Chongde;Jun, Huang;Zhou, Rongqing;Shi, Bi
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1168-1177
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    • 2018
  • The aim of this study was to investigate the microbial community of three tannery wastewater treatment plants (WWTPs) involved in nitrification by Illumina MiSeq sequencing. The results showed that highly diverse communities were present in tannery wastewater. A total of six phyla, including Proteobacteria (37-41%), Bacteroidetes (6.04-16.80), Planctomycetes (3.65-16.55), Chloroflexi (2.51-11.48), Actinobacteria (1.91-9.21), and Acidobacteria (3.04-6.20), were identified as the main phyla, and Proteobacteria dominated in all the samples. Within Proteobacteria, Beta-proteobacteria was the most abundant class, with the sequence percentages ranging from 9.66% to 17.44%. Analysis of the community at the genus level suggested that Thauera, Gp4, Ignavibacterium, Phycisphaera, and Arenimonas were the core genera shared by at least two tannery WWTPs. A detailed analysis of the abundance of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) indicated that Nitrosospira, Nitrosomonas, and Nitrospira were the main AOB and NOB in tannery wastewater, respectively, which exhibited relatively high abundance in all samples. In addition, real-time quantitative PCR was conducted to validate the results by quantifying the abundance of the AOB and total bacteria, and similar results were obtained. Overall, the results presented in this study may provide new insights into our understanding of key microorganisms and the entire community of tannery wastewater and contribute to improving the nitrogen removal efficiency.

Effects of Antibiotic Growth Promoter and Characterization of Ecological Succession in Swine Gut Microbiota

  • Unno, Tatsuya;Kim, Jungman;Guevarra, Robin B.;Nguyen, Son G.
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.431-438
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    • 2015
  • Ever since the ban on antibiotic growth promoters (AGPs), the livestock death rate has increased owing to pathogenic bacterial infections. There is a need of developing AGP alternatives; however, the mechanisms by which AGP enhances livestock growth performance are not clearly understood. In this study, we fed 3-week-old swine for 9 weeks with and without AGPs containing chlortetracycline, sulfathiazole, and penicillin to investigate the effects of AGPs on swine gut microbiota. Microbial community analysis was done based on bacterial 16S rRNA genes using MiSeq. The use of AGP showed no growth promoting effect, but inhibited the growth of potential pathogens during the early growth stage. Our results showed the significant increase in species richness after the stabilization of gut microbiota during the post-weaning period (4-week-old). Moreover, the swine gut microbiota was divided into four clusters based on the distribution of operational taxonomic units, which was significantly correlated to the swine weight regardless of AGP treatments. Taxonomic abundance analysis indicated a negative correlation between host weight and the abundance of the family Prevotellaceae species, but showed positive correlation to the abundance of the family Spirochaetaceae, Clostridiaceae_1, and Peptostreptococcaeae species. Although no growth performance enhancement was observed, the use of AGP inhibited the potential pathogens in the early growth stage of swine. In addition, our results indicated the ecological succession of swine gut microbiota according to swine weight. Here, we present a characterization of swine gut microbiota with respect to the effects of AGPs on growth performance.

Fungal Community Analyses of Endophytic Fungi from Two Oak Species, Quercus mongolica and Quercus serrata, in Korea

  • Nguyen, Manh Ha;Shin, Keum Chul;Lee, Jong Kyu
    • Mycobiology
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    • v.49 no.4
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    • pp.385-395
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    • 2021
  • Fungal endophytes have been recorded in various plant species with a richness of diversity, and their presence plays an essential role in host plant protection against biotic and abiotic stresses. This study applied the Illumina MiSeq sequencing platform based on the amplification of fungal ribosomal ITS2 region to analyze fungal endophytic communities of two oak species (Quercus mongolica and Q. serrata) with different oak wilt disease susceptibilities in Korea. The results showed a total of 230,768 sequencing reads were obtained and clustered at a 97% similarity threshold into 709 operational taxonomic units (OTUs). The OTUs of Q. serrata were higher than that of Q. mongolica with the number of 617 OTUs and 512 OTUs, respectively. Shannon index also showed that Q. serrata had a significantly higher level of fungal diversity than Q. mongolica. Total of OTUs were assigned into 5 fungal phyla, 17 classes, 60 orders, 133 families, 195 genera, and 280 species. Ascomycota was the dominant phylum with 75.11% relative abundance, followed by Basidiomycota with 5.28%. Leptosillia, Aureobasidium and Acanthostigma were the most abundant genera detected in Q. serrata with the average relative abundance of 2.85, 2.76, and 2.19%, respectively. On the other hand, Peltaster, Cladosporium and Monochaetia were the most common genera detected in Q. mongolica with the average relative abundance of 4.83, 3.03, and 2.87%, respectively. Our results indicated that fungal endophytic communities were significantly different between two oak species and these differences could influence responses of host trees to oak wilt disease caused by Raffaelea quercus-mongolicae.

Expression profiles of microRNAs in skeletal muscle of sheep by deep sequencing

  • Liu, Zhijin;Li, Cunyuan;Li, Xiaoyue;Yao, Yang;Ni, Wei;Zhang, Xiangyu;Cao, Yang;Hazi, Wureli;Wang, Dawei;Quan, Renzhe;Yu, Shuting;Wu, Yuyu;Niu, Songmin;Cui, Yulong;Khan, Yaseen;Hu, Shengwei
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.757-766
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    • 2019
  • Objective: MicroRNAs are a class of endogenous small regulatory RNAs that regulate cell proliferation, differentiation and apoptosis. Recent studies on miRNAs are mainly focused on mice, human and pig. However, the studies on miRNAs in skeletal muscle of sheep are not comprehensive. Methods: RNA-seq technology was used to perform genomic analysis of miRNAs in prenatal and postnatal skeletal muscle of sheep. Targeted genes were predicted using miRanda software and miRNA-mRNA interactions were verified by quantitative real-time polymerase chain reaction. To further investigate the function of miRNAs, candidate targeted genes were enriched for analysis using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment. Results: The results showed total of 1,086 known miRNAs and 40 new candidate miRNAs were detected in prenatal and postnatal skeletal muscle of sheep. In addition, 345 miRNAs (151 up-regulated, 94 down-regulated) were differentially expressed. Moreover, miRanda software was performed to predict targeted genes of miRNAs, resulting in a total of 2,833 predicted targets, especially miR-381 which targeted multiple muscle-related mRNAs. Furthermore, GO and KEGG pathway analysis confirmed that targeted genes of miRNAs were involved in development of skeletal muscles. Conclusion: This study supplements the miRNA database of sheep, which provides valuable information for further study of the biological function of miRNAs in sheep skeletal muscle.

Integrated mRNA and miRNA profile expression in livers of Jinhua and Landrace pigs

  • Huang, Minjie;Chen, Lixing;Shen, Yifei;Chen, Jiucheng;Guo, Xiaoling;Xu, Ningying
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.10
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    • pp.1483-1490
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    • 2019
  • Objective: To explore the molecular mechanisms of fat metabolism and deposition in pigs, an experiment was conducted to identify hepatic mRNAs and miRNAs expression and determine the potential interaction of them in two phenotypically extreme pig breeds. Methods: mRNA and miRNA profiling of liver from 70-day Jinhua (JH) and Landrace (LD) pigs were performed using RNA sequencing. Blood samples were taken to detect results of serum biochemistry. Bioinformatics analysis were applied to construct differentially expressed miRNA-mRNA network. Results: Serum total triiodothyronine and total thyroxine were significantly lower in Jinhua pigs, but the content of serum total cholesterol (TCH) and low-density lipoprotein cholesterol were strikingly higher. A total of 467 differentially expressed genes (DEGs) and 35 differentially expressed miRNAs (DE miRNAs) were identified between JH and LD groups. Gene ontology analysis suggested that DEGs were involved in oxidation-reduction, lipid biosynthetic and lipid metabolism process. Interaction network of DEGs and DE miRNAs were constructed, according to target prediction results. Conclusion: We generated transcriptome and miRNAome profiles of liver from JH and LD pig breeds which represent distinguishing phenotypes of growth and metabolism. The potential miRNA-mRNA interaction networks may provide a comprehensive understanding in the mechanism of lipid metabolism. These results serve as a basis for further investigation on biological functions of miRNAs in the porcine liver.

Rhizospheric fungi of Panax notoginseng: diversity and antagonism to host phytopathogens

  • Miao, Cui-Ping;Mi, Qi-Li;Qiao, Xin-Guo;Zheng, You-Kun;Chen, You-Wei;Xu, Li-Hua;Guan, Hui-Lin;Zhao, Li-Xing
    • Journal of Ginseng Research
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    • v.40 no.2
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    • pp.127-134
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    • 2016
  • Background: Rhizospheric fungi play an essential role in the plantesoil ecosystem, affecting plant growth and health. In this study, we evaluated the fungal diversity in the rhizosphere soil of 2-yr-old healthy Panax notoginseng cultivated in Wenshan, China. Methods: Culture-independent Illumina MiSeq and culture-dependent techniques, combining molecular and morphological characteristics, were used to analyze the rhizospheric fungal diversity. A diffusion test was used to challenge the phytopathogens of P. notoginseng. Results: A total of 16,130 paired-end reads of the nuclear ribosomal internal transcribed spacer 2 were generated and clustered into 860 operational taxonomic units at 97% sequence similarity. All the operational taxonomic units were assigned to five phyla and 79 genera. Zygomycota (46.2%) and Ascomycota (37.8%) were the dominant taxa; Mortierella and unclassified Mortierellales accounted for a large proportion (44.9%) at genus level. The relative abundance of Fusarium and Phoma sequenceswas high, accounting for 12.9% and 5.5%, respectively. In total,113 fungal isolates were isolated from rhizosphere soil. They were assigned to five classes, eight orders (except for an Incertae sedis), 26 genera, and 43 species based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer. Fusarium was the most isolated genus with six species (24 isolates, 21.2%). The abundance of Phoma was also relatively high (8.0%). Thirteen isolates displayed antimicrobial activity against at least one test fungus. Conclusion: Our results suggest that diverse fungi including potential pathogenic ones exist in the rhizosphere soil of 2-yr-old P. notoginseng and that antagonistic isolates may be useful for biological control of pathogens.

Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing (차세대염기서열분석법을 이용한 HLA-A, -B 그리고 -DR 형별 분석법 개발)

  • Seo, Dong Hee;Lee, Jeong Min;Park, Mi Ok;Lee, Hyun Ju;Moon, Seo Yoon;Oh, Mijin;Kim, So Young;Lee, Sang-Heon;Hyeong, Ki-Eun;Hu, Hae-Jin;Cho, Dae-Yeon
    • The Korean Journal of Blood Transfusion
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    • v.29 no.3
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    • pp.310-319
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    • 2018
  • Background: Research on next-generation sequencing (NGS)-based HLA typing is active. To resolve the phase ambiguity and long turn-around-time of conventional high resolution HLA typing, this study developed a NGS-based high resolution HLA typing method that can handle large-scale samples within an efficient testing time. Methods: For HLA NGS, the condition of nucleic acid extraction, library construction, PCR mechanism, and HLA typing with bioinformatics were developed. To confirm the accuracy of the NGS-based HLA typing method, the results of 192 samples HLA typed by SSOP and 28 samples typed by SBT compared to NGS-based HLA-A, -B and -DR typing. Results: DNA library construction through two-step PCR, NGS sequencing with MiSeq (Illumina Inc., San Diego, USA), and the data analysis platform were established. NGS-based HLA typing results were compatible with known HLA types from 220 blood samples. Conclusion: The NSG-based HLA typing method could handle large volume samples with high-throughput. Therefore, it would be useful for HLA typing of bone marrow donation volunteers.

Assessment Tools of Sensory Processing for Preschool-Aged Children: A Systematic Review (학령전기 아동을 위한 감각처리 평가도구의 체계적 고찰)

  • Kim, Myung-Hee;Shin, Hyeon-Hui;Kim, Kyeong-Mi
    • The Journal of Korean Academy of Sensory Integration
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    • v.13 no.1
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    • pp.1-12
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    • 2015
  • Objective : The purpose of the study was to investigate the characteristics and psychometric properties of assessments which were developed to measure sensory processing functions of preschool-aged children. Method : Comprehensive search for assessments measuring sensory processing functions was administered by using multiple electronic databases, including PubMed, Cochrane, CINAHL and PsycINFO. The titles and abstracts from the retrieved papers were screened. Then assessment tools that met all of the inclusion criteria were selected. Results : Thirty-eight assessments were identified. There were eight assessment tools that met the pre-defined inclusion criteria. Sensory Processing Measure-Preschool (SPM-P) was verified to be the most reliable and valid assessment tool, whereas Sensory Experiences Questionnaire (SEQ), Sense & Self-Regulation Checklist (SSRC), and Sensory Processing Scale Assessment (SPSA) showed inadequate reliability and validity. Conclusion : Based on the results from this systematic review, it would be useful for clinicians and researchers to select assessment tools of sensory processing for preschool-aged children.

Complete genome sequence of Comamonas sp. NLF-7-7 isolated from biofilter of wastewater treatment plant (폐수처리장의 바이오 필터로부터 분리된 Comamonas sp. NLF-7-7 균주의 유전체 염기서열 해독)

  • Kim, Dong-Hyun;Han, Kook-Il;Kwon, Hae Jun;Kim, Mi Gyeong;Kim, Young Guk;Choi, Doo Ho;Lee, Keun Chul;Suh, Min Kuk;Kim, Han Sol;Lee, Jung-Sook;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.309-312
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    • 2019
  • Comamonas sp. NLF-7-7 was isolated from biofilter of wastewater treatment plant. The whole-genome sequence of Comamonas sp. NLF-7-7 was analyzed using the PacBio RS II and Illumina HiSeqXten platform. The genome comprises a 3,333,437 bp chromosome with a G + C content of 68.04%, 3,197 total genes, 9 rRNA genes, and 49 tRNA genes. This genome contained pollutants degradation and floc forming genes such as sulfur oxidization pathway (SoxY, SoxZ, SoxA, and SoxB) and floc forming pathway (EpsG, EpsE, EpsF, EpsG, EpsL, and glycosyltransferase), respectively. The Comamonas sp. NLF-7-7 can be used to the purification of wastewater.

Transcriptome and Flower Color Related Gene Analysis in Angelica gigas Nakai Using RNA-Seq (RNA-seq을 이용한 참당귀의 전사체 분석과 꽃 색 관련 유전자 분석)

  • Kim, Nam Su;Jung, Dae Hui;Park, Hong Woo;Park, Yun mi;Jeon, Kwon Seok;Kim, Mahn Jo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.73-73
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    • 2019
  • Angelica gigas Nakai (Korean danggui), a member of the Umbelliferae family, is a Korean traditional medicinal plant whose roots have been used for treating gynecological diseases. Transcriptomics is the study of the transcriptome, which is the complete set of RNA transcripts that are produced by the genome, using high-throughput methods, such as microarray analysis. In this study, transcriptome analysis of A.gigas Nakai was carried out. Transcriptome sequencing and assembly was carried out by using Illumina Hiseq 2500, Velvet and Oases. A total of 109,591,555 clean reads of A. gigas Nakai was obtained after trimming adaptors. The obtained reads were assembled with an average length of 1,154 bp, a maximum length of 13,166 bp, a minimum length of 200 pb, and N50 of 1,635 bp. Functional annotation and classification was performed using NCBI NR, InterprotScan, KOG, KEGG and GO. Candidate genes for phenylpropanoid biosynthesis were obtanied from A.gigas transcriptome and the genes and its proteins were confirmed through the NCBI homology BLAST searches, revealing high identity with other othologous genes and proteins from various plants pecies. In RNA sequencing analysis using an Illumina Next-Seq2500 sequencer, we identified a total 94,930 transcripts and annotated 71,281 transcripts, which provide basic information for further research in A.gigas Nakai. Our transcriptome data reveal that several differentially expressed genes related to flower color in A.gigas Nakai. The results of this research provide comprehensive information on the A.gigas Nakai genome and enhance our understanding of the flower color related gene pathways in this plant.

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