• Title/Summary/Keyword: Next Generation Sequencing

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Microbes causing abnormal Takju fermentation in traditional wheat-based Nuruk: A case study (밀누룩으로 제조한 전통탁주의 이상발효 원인 규명: 사례분석)

  • Yun, Jeonghyun;Lee, Jang-Eun
    • Korean Journal of Food Science and Technology
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    • v.54 no.3
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    • pp.343-350
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    • 2022
  • In this article, we report a case study on the cause of abnormal fermentation of Takju based on a microbial community analysis of wheat-based Nuruk and Takju using next-generation sequencing. The microbial community showed notable changes during alcoholic fermentation. Members of the genera Aspergillus and Staphylococcus were dominant in Nuruk, whereas Saccharomyces, Pediococcus, Bacillus, Lactobacillus, and Enterobacter were predominant during Takju fermentation. In particular, members of the Enterobacter and Bacillus genera that were present were identified as the opportunistic pathogens B. wiedmannii and E. cloacae. The relative abundance of two species increased in Mitsul-3, wherein the concentration of organic acid rapidly decreased, and in Deotsul-1, in which raw materials were added. Furthermore, the relative abundance of lactic acid bacteria (LAB) in both the Nuruk and Takju was very low. Therefore, this result showed that the abnormal proliferation of E. cloacae and B. wiedmannii in Takju was due to the absence of LAB.

Crop Effects on Soil Microorganism Activity and Community Composition in the Agricultural Environment (농경지에서 재배작물이 토양미생물활성 및 군집구성에 미치는 영향)

  • Bak, Gyeryeong;Lee, Jeong-Tae;Jee, Samnyu
    • Journal of Environmental Science International
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    • v.30 no.5
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    • pp.379-389
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    • 2021
  • Soil microorganism activity in an agricultural field is affected by various factors including climate conditions, soil chemical properties, and crop cultivation. In this study, we elucidate the correlation between microorganism activity and agricultural environment factors using the dehydrogenase activity (DHA) value, which is one of the indicators of soil microbial activity. As a result, the various factors noted above were related to the DHA value. Annual rainfall, soil Mg2+, bacterial and fungal diversities, types of crops, developmental stages, seasons, and cultivation status were highly correlated with the DHA value. Furthermore, next-generation sequencing (NGS) analysis was used to identify that the type of crop affected soil microbial compositions of both bacteria and fungi. Soil used for soybean cultivation showed the highest relative abundance for Verrucomicrobia, Planctomycetes, and Acidobacteria but Actinobacteria and Firmicutes had the lowest relative abundance. In the case of soil used for potato cultivation, Actinobacteria had the highest relative abundance but Proteobacteria had the lowest relative abundance. Armatimonadetes showed the highest relative abundance in soil used for cabbage cultivation. Among the fungal communities, Mortierellomycota had the highest relative abundance for soybean cultivation but the lowest relative abundance for cabbage cultivation; further, Rozellomycota, Chytridiomycota, and Cercozoa had the highest relative abundance for cabbage cultivation. Basidiomycota had the highest relative abundance for potato cultivation but the lowest relative abundance for soybean cultivation.

Effects of lactic acid bacteria fermented feed and three types of lactic acid bacteria (L. plantarum, L. acidophilus, B. animalis) on intestinal microbiota and T cell polarization (Th1, Th2, Th17, Treg) in the intestinal lymph nodes and spleens of rats

  • Da Yoon, Yu;Sang-Hyon, Oh;In Sung, Kim;Gwang Il, Kim;Jeong A, Kim;Yang Soo, Moon;Jae Cheol, Jang;Sang Suk, Lee;Jong Hyun, Jung;Hwa Chun, Park;Kwang Keun, Cho
    • Animal Bioscience
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    • v.36 no.1
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    • pp.156-166
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    • 2023
  • Objective: In this study, we investigated the effects of Rubus coreanus-derived lactic acid bacteria (LAB) fermented feed (RC-LAB fermented feed) and three types of LAB (Lactobacillus plantarum, Lactobacillus acidophilus, Bifidobacterium animalis) on the expression of transcription factors and cytokines in Th1, Th2, Th17, and Treg cells in the intestinal lymph nodes and spleens of rats. In addition, the effect on intestinal microbiota composition and body weight was investigated. Methods: Five-week-old male rats were assigned to five treatments and eight replicates. The expression of transcription factors and cytokines of Th1, Th2, Th17, and Treg cells in the intestinal lymph nodes and spleens was analyzed using real-time reverse transcriptase polymerase chain reaction assays. Intestinal tract microbiota compositions were analyzed by next-generation sequencing and quantitative polymerase chain reaction assays. Results: RC-LAB fermented feed and three types of LAB increased the expression of transcription factors and cytokines in Th1, Treg cells and Galectin-9, but decreased in Th2 and Th17 cells. In addition, the intestinal microbiota composition changed, the body weight and Firmicutes to Bacteroidetes (F/B) ratio decreased, and the relative abundance of LAB increased. Conclusion: LAB fermented feed and three types of LAB showed an immune modulation effect by inducing T cell polarization and increased LAB in the intestinal microbiota.

Identification of unbalanced complex chromosomal rearrangements in IVF-derived embryos during NGS analysis of preimplantation genetic testing: A case report

  • Yu, Eun Jeong;Kim, Min Jee;Park, Eun A;Hong, Ye Seul;Park, Sun Ok;Park, Sang-Hee;Lee, Yu Bin;Yoon, Tae Ki;Kang, Inn Soo
    • Journal of Genetic Medicine
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    • v.19 no.1
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    • pp.14-21
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    • 2022
  • Complex chromosome rearrangements (CCRs) are structural chromosomal rearrangements involving at least three chromosomes and more than two breakpoints. CCR carriers are generally phenotypically normal but related to higher risk of recurrent miscarriage and having abnormal offspring with congenital anomalies. However, most of CCR carriers are not aware of their condition until genetic analysis of either abortus or affected baby or parental karyotyping is performed. Herein, we present the case that CCR carrier patients can be identified by preimplantation genetic testing of preimplantation embryos. An infertile male patient with severe oligoasthenoteratozoospermia was diagnosed balanced reciprocal translocation, 46,XY,t(3;11) (p26;p14) at first. After attempting the first preimplantation genetic testing for structural rearrangement (PGT-SR) cycle, we found the recurrent segmental gain or loss on 21q21.3-q22.3 of five out of nine embryos. As a result of karyotype re-analysis, the patient's karyotype showed a balanced CCR involving chromosomes 3, 11, and 21 with three breakpoints 3p26, 11p14, and 21q21. The patient underwent two PGT-SR cycles, and a pregnancy was established after the transfer of an euploid embryo in the second cycle. Amniocentesis confirmed that the baby carried normal karyotype without mosaicism. At 37 weeks gestation, a healthy girl weighting 3,050 g was born.

Lophomonas blattarum-like organism in bronchoalveolar lavage from a pneumonia patient: current diagnostic scheme and polymerase chain reaction can lead to false-positive results

  • Moses Lee;Sang Mee Hwang;Jong Sun Park;Jae Hyeon Park;Jeong Su Park
    • Parasites, Hosts and Diseases
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    • v.61 no.2
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    • pp.202-209
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    • 2023
  • Lophomonas blattarum is an anaerobic protozoan living in the intestine of cockroaches and house dust mites, with ultramicroscopic characteristics such as the presence of a parabasal body, axial filament, and absence of mitochondria. More than 200 cases of Lophomonas infection of the respiratory tract have been reported worldwide. However, the current diagnosis of such infection depends only on light microscopic morphological findings from respiratory secretions. In this study, we attempted to provide more robust evidence of protozoal infection in an immunocompromised patient with atypical pneumonia, positive for Lophomonas-like protozoal cell forms. A direct search of bronchoalveolar lavage fluid via polymerase chain reaction (PCR), transmission electron microscopy (TEM), and metagenomic next-generation sequencing did not prove the presence of protozoal infection. PCR results were not validated with sufficient rigor, while de novo assembly and taxonomic classification results did not confirm the presence of an unidentified pathogen. The TEM results implied that such protozoal forms in light microscopy are actually non-detached ciliated epithelial cells. After ruling out infectious causes, the patient's final diagnosis was drug-induced pneumonitis. These findings underscore the lack of validation in the previously utilized diagnostic methods, and more evidence in the presence of L. blattarum is required to further prove its pathogenicity.

Diagnosing Balamuthia mandrillaris amebic meningoencephalitis in a 64-year-old woman from the Southwest of China

  • Suhua Yao;Xiaoting Chen;Lian Qian;Shizheng Sun;Chunjing Zhao;Zongkai Bai;Zhaofang Chen;Youcong Wu
    • Parasites, Hosts and Diseases
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    • v.61 no.2
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    • pp.183-193
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    • 2023
  • Balamuthia mandrillaris amebic encephalitis (BAE) can cause a fatal condition if diagnosis is delayed or effective treatment is lacking. Patients with BAE have been previously reported in 12 provinces of China, with skin lesions being the primary symptom and encephalitis developing after several years. However, a significantly lower number of cases has been reported in Southwest China. Here we report an aggressive BAE case of a 64-year-old woman farmer with a history of skin lesions on her left hand. She was admitted to our hospital due to symptoms of dizziness, headache, cough, vomiting, and gait instability. She was initially diagnosed with syphilitic meningoencephalitis and received a variety of empirical treatment that failed to improve her symptoms. Finally, she was diagnosed with BAE combined with amebic pneumonia using next-generation sequencing (NGS), qRT-PCR, sequence analysis, and imaging studies. She died approximately 3 weeks after the onset. This case highlights that the rapid development of encephalitis can be a prominent clinical manifestation of Balamuthia mandrillaris infection.

Characterization of a Potential Probiotic Lactiplantibacillus plantarum LRCC5310 by Comparative Genomic Analysis and its Vitamin B6 Production Ability

  • Yunjeong Lee;Nattira Jaikwang;Seong keun Kim;Jiseon Jeong;Ampaitip Sukhoom;Jong-Hwa Kim;Wonyong Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.5
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    • pp.644-655
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    • 2023
  • Safety assessment and functional analysis of probiotic candidates are important for their industrial applications. Lactiplantibacillus plantarum is one of the most widely recognized probiotic strains. In this study we aimed to determine the functional genes of L. plantarum LRCC5310, isolated from kimchi, using next-generation, whole-genome sequencing analysis. Genes were annotated using the Rapid Annotations using Subsystems Technology (RAST) server and the National Center for Biotechnology Information (NCBI) pipelines to establish the strain's probiotic potential. Phylogenetic analysis of L. plantarum LRCC5310 and related strains showed that LRCC5310 belonged to L. plantarum. However, comparative analysis revealed genetic differences between L. plantarum strains. Carbon metabolic pathway analysis based on the Kyoto Encyclopedia of Genes and Genomes database showed that L. plantarum LRCC5310 is a homofermentative bacterium. Furthermore, gene annotation results indicated that the L. plantarum LRCC5310 genome encodes an almost complete vitamin B6 biosynthetic pathway. Among five L. plantarum strains, including L. plantarum ATCC 14917T , L. plantarum LRCC5310 detected the highest concentration of pyridoxal 5'-phosphate with 88.08 ± 0.67 nM in MRS broth. These results indicated that L. plantarum LRCC5310 could be used as a functional probiotic for vitamin B6 supplementation.

COVID-19 progression towards ARDS: a genome wide study reveals host factors underlying critical COVID-19

  • Shama Mujawar;Gayatri Patil;Srushti Suthar;Tanuja Shendkar;Vaishnavi Gangadhar
    • Genomics & Informatics
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    • v.21 no.2
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    • pp.16.1-16.14
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    • 2023
  • Coronavirus disease 2019 (COVID-19) is a viral infection produced by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus epidemic, which was declared a global pandemic in March 2020. The World Health Organization has recorded around 43.3 billion cases and 59.4 million casualties to date, posing a severe threat to global health. Severe COVID-19 indicates viral pneumonia caused by the SARS-CoV-2 infections, which can induce fatal consequences, including acute respiratory distress syndrome (ARDS). The purpose of this research is to better understand the COVID-19 and ARDS pathways, as well as to find targeted single nucleotide polymorphism. To accomplish this, we retrieved over 100 patients' samples from the Sequence Read Archive, National Center for Biotechnology Information. These sequences were processed through the Galaxy server next generation sequencing pipeline for variant analysis and then visualized in the Integrative Genomics Viewer, and performed statistical analysis using t-tests and Bonferroni correction, where six major genes were identified as DNAH7, CLUAP1, PPA2, PAPSS1, TLR4, and IFITM3. Furthermore, a complete understanding of the genomes of COVID-19-related ARDS will aid in the early identification and treatment of target proteins. Finally, the discovery of novel therapeutics based on discovered proteins can assist to slow the progression of ARDS and lower fatality rates.

Effective microbial molecular diagnosis of periodontitis-related pathogen Porphyromonas gingivalis from salivary samples using rgpA gene

  • Jinuk Jeong;Yunseok Oh;Junhyeon Jeon;Dong-Heon Baek;Dong Hee Kim;Kornsorn Srikulnath;Kyudong Han
    • Genomics & Informatics
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    • v.21 no.1
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    • pp.13.1-13.8
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    • 2023
  • Importance of accurate molecular diagnosis and quantification of particular disease-related pathogenic microorganisms is highlighted as an introductory step to prevent and care for diseases. In this study, we designed a primer/probe set for quantitative real-time polymerase chain reaction (qRT-PCR) targeting rgpA gene, known as the specific virulence factor of periodontitis-related pathogenic bacteria 'Porphyromonas gingivalis', and evaluated its diagnostic efficiency by detecting and quantifying relative bacterial load of P. gingivalis within saliva samples collected from clinical subjects. As a result of qRT-PCR, we confirmed that relative bacterial load of P. gingivalis was detected and quantified within all samples of positive control and periodontitis groups. On the contrary, negative results were confirmed in both negative control and healthy groups. Additionally, as a result of comparison with next-generation sequencing (NGS)-based 16S metagenome profiling data, we confirmed relative bacterial load of P. gingivalis, which was not identified on bacterial classification table created through 16S microbiome analysis, in qRT-PCR results. It showed that an approach to quantifying specific microorganisms by applying qRT-PCR method could solve microbial misclassification issues at species level of an NGS-based 16S microbiome study. In this respect, we suggest that P. gingivalis-specific primer/probe set introduced in present study has efficient applicability in various oral healthcare industries, including periodontitis-related microbial molecular diagnosis field.

State of the art on the physical mapping of the Y-chromosome in the Bovidae and comparison with other species - A review

  • Rossetti, Cristina;Genualdo, Viviana;Incarnato, Domenico;Mottola, Filomena;Perucatti, Angela;Pauciullo, Alfredo
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1289-1302
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    • 2022
  • The next generation sequencing has significantly contributed to clarify the genome structure of many species of zootechnical interest. However, to date, some portions of the genome, especially those linked to a heterogametic nature such as the Y chromosome, are difficult to assemble and many gaps are still present. It is well known that the fluorescence in situ hybridization (FISH) is an excellent tool for identifying genes unequivocably mapped on chromosomes. Therefore, FISH can contribute to the localization of unplaced genome sequences, as well as to correct assembly errors generated by comparative bioinformatics. To this end, it is necessary to have starting points; therefore, in this study, we reviewed the physically mapped genes on the Y chromosome of cattle, buffalo, sheep, goats, pigs, horses and alpacas. A total of 208 loci were currently mapped by FISH. 89 were located in the male-specific region of the Y chromosome (MSY) and 119 were identified in the pseudoautosomal region (PAR). The loci reported in MSY and PAR were respectively: 18 and 25 in Bos taurus, 5 and 7 in Bubalus bubalis, 5 and 24 in Ovis aries, 5 and 19 in Capra hircus, 10 and 16 in Sus scrofa, 46 and 18 in Equus caballus. While in Vicugna pacos only 10 loci are reported in the PAR region. The correct knowledge and assembly of all genome sequences, including those of genes mapped on the Y chromosome, will help to elucidate their biological processes, as well as to discover and exploit potentially epistasis effects useful for selection breeding programs.