scholarly journals Fate of Functional Bacterial and Eukaryotic Community Regulated by Earthworms during Vermicomposting of Dewatered Sludge, Studies Based on the 16S rDNA and 18S rDNA Sequencing of Active Cells

2021 ◽  
Vol 18 (18) ◽  
pp. 9713
Author(s):  
Jun Yang ◽  
Kui Huang ◽  
Lansheng Peng ◽  
Jianhui Li ◽  
Aozhan Liu
Keyword(s):  
16S Rdna ◽  
18S Rdna ◽  
High Throughput Sequencing ◽  
Propidium Monoazide ◽  
Eukaryotic Community ◽  
Rdna Sequencing ◽  
Stable Product ◽  
Active Bacteria ◽  
Dewatered Sludge ◽  
And Control

DNA sequencing of active cells involved in vermicomposting can clarify the roles of earthworms in regulating functional microorganisms. This study aimed to investigate the effect of earthworms on functional microbial communities in sludge by comparing biodegradation treatments with and without earthworms. PCR and high throughput sequencing based on pretreatment of propidium monoazide (PMA) were used to detect the changes in active bacterial 16S rDNA and eukaryotic 18S rDNA during vermicomposting. The results showed that the nitrate in sludge vermicomposting and control were significantly different from day 10, with a more stable product at day 30 of vermicomposting. Compared with the control, the Shannon indexes of active bacteria and eukaryotes decreased by 1.9% and 31.1%, respectively, in sludge vermicompost. Moreover, Proteobacteria (36.2%), Actinobacteria (25.6%), and eukaryotic Cryptomycota (80.3%) were activated in the sludge vermicompost. In contrast, the control had Proteobacteria (44.8%), Bacteroidetes (14.2%), Cryptomycota (50.00%), and Arthropoda (36.59%). Network analysis showed that environmental factors had different correlations between active bacterial and eukaryotic community structures. This study suggests that earthworms can decrease the diversity of bacterial and eukaryotic communities, forming a specific-functional microbial community and thus accelerating organic matter decomposition during vermicomposting of dewatered sludge.

scholarly journals Vaginal Microbiota Diversity of Patients with Embryonic Miscarriage by Using 16S rDNA High-Throughput Sequencing

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Linfen Xu ◽  
Lingna Huang ◽  
Chengying Lian ◽  
Huili Xue ◽  
Yanfang Lu ◽  
...  
Keyword(s):  
Pregnant Women ◽  
16S Rdna ◽  
High Throughput Sequencing ◽  
Interleukin 2 ◽  
Life Quality ◽  
Control Group ◽  
Operational Taxonomic Units ◽  
Causative Relationship ◽  
Rdna Sequencing ◽  
And Control

Embryonic miscarriage severely affects the life quality and physical and mental state of pregnant women. However, the detailed mechanism underlying embryonic miscarriage is not fully understood. This study is aimed at analyzing embryonic miscarriage. We collected samples from 25 normal pregnant women and 25 embryonic miscarriage patients of similar age to analyze microbiota isolated from the vagina. Crude examination of the vagina isolates showed that compared with the control group, 80% of the embryonic miscarriage group contained a significantly lower number of Lactobacillus, the major healthy microbe in the vagina. Furthermore, the levels of Th1 and Th2 secreted cytokine interleukin 2 (IL-2) and interleukin10 (IL-10), respectively, were examined. Results showed that the IL2 level was higher, and IL10 level was lower in the embryonic miscarriage group than in the control group, whereas the IL2/IL10 level was higher in the embryonic miscarriage group than in the control group. This finding suggested that the immune response was suppressed in the embryonic miscarriage group. To further dissect the microbiota of the vagina in the two groups, 16S rDNA sequencing was performed. Bioinformatics analysis showed that 1096 and 998 overlapped operational taxonomic units were identified from the embryonic miscarriage and control groups, respectively. At the genus level, the relative abundance of Fam_Finegoldia, Lac_Coprococcus_3, and Lac_Roseburia significantly differed in the embryonic miscarriage group. Overall, our analyses provided potential biomarkers for embryonic miscarriage and elucidated the causative relationship between microbiota and immune responses and may enable the possible diagnosis and therapeutics of early pregnancy loss.

scholarly journals High-throughput sequencing revealed differences of microbial community structure and diversity between healthy and diseased Caulerpa lentillifera

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Zhourui Liang ◽  
Fuli Liu ◽  
Wenjun Wang ◽  
Pengyan Zhang ◽  
Xiutao Sun ◽  
...  
Keyword(s):  
Information Processing ◽  
Community Structure ◽  
Microbial Community ◽  
16S Rdna ◽  
High Throughput ◽  
Microbial Community Structure ◽  
18S Rdna ◽  
High Throughput Sequencing ◽  
Rdna Sequencing ◽  
Caulerpa Lentillifera

Abstract Background Caulerpa lentillifera is one of the most important economic green macroalgae in the world. Increasing demand for consumption has led to the commercial cultivation of C. lentillifera in Japan and Vietnam in recent decades. Concomitant with the increase of C. lentillifera cultivation is a rise in disease. We hypothesise that epiphytes or other microorganisms outbreak at the C. lentillifera farm may be an important factor contributing to disease in C. lentillifera. The main aims are obtaining differences in the microbial community structure and diversity between healthy and diseased C. lentillifera and key epiphytes and other microorganisms affecting the differences through the results of high-throughput sequencing and bioinformatics analysis in the present study. Results A total of 14,050, 2479, and 941 operational taxonomic units (OTUs) were obtained from all samples using 16S rDNA, 18S rDNA, and internal transcribed spacer (ITS) high-throughput sequencing, respectively. 16S rDNA sequencing and 18S rDNA sequencing showed that microbial community diversity was higher in diseased C. lentillifera than in healthy C. lentillifera. Both PCoA results and UPGMA results indicated that the healthy and diseased algae samples have characteristically different microbial communities. The predominant prokaryotic phyla were Proteobacteria, Planctomycetes, Bacteroidetes, Cyanobacteria, Acidobacteria, Acidobacteria and Parcubacteria in all sequences. Chlorophyta was the most abundant eukaryotic phylum followed by Bacillariophyta based on 18S rDNA sequencing. Ascomycota was the dominant fungal phylum detected in healthy C. lentillifera based on ITS sequencing, whereas fungi was rare in diseased C. lentillifera, suggesting that Ascomycota was probably fungal endosymbiont in healthy C. lentillifera. There was a significantly higher abundance of Bacteroidetes, Cyanobacteria, Bacillariophyta, Ulvales and Tetraselmis in diseased C. lentillifera than in healthy C. lentillifera. Disease outbreaks significantly change carbohydrate metabolism, environmental information processing and genetic information processing of prokaryotic communities in C. lentillifera through predicted functional analyses using the Tax4Fun tool. Conclusions Bacteroidetes, Cyanobacteria, Bacillariophyta, Ulvales and Tetraselmis outbreak at the C. lentillifera farm sites was an important factor contributing to disease in C. lentillifera.

scholarly journals Study on the Characteristics of Gut Microbiota in Chronic Hepatitis B Patients With Damp Heat Syndrome and Liver Depression and Spleen Deficiency Syndrome

2020 ◽  
Author(s):  
Kaiping Jiang ◽  
Qunfang Jiang ◽  
Xiaoai Mo ◽  
Jianhong Li ◽  
Hongtao Hu ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
16S Rdna ◽  
Relative Abundance ◽  
High Throughput Sequencing ◽  
Deficiency Syndrome ◽  
Spleen Deficiency ◽  
Rdna Sequencing ◽  
Significant Difference ◽  
Heat Syndrome ◽  
Spleen Deficiency Syndrome

Abstract Background: To explore the potential biological characteristics of traditional Chinese medicine(TCM) syndromes in CHB patients is of great significance to improve the level of TCM treatment.This study was designed to explore the characteristics of gut microbiota in CHB patients with two most common TCM Syndromes.Methods: According to the diagnostic criteria of Western medicine, TCM and screening exclusion criteria, 65 cases of CHB patients with damp heat syndrome and 28 cases of CHB patients with liver depression and spleen deficiency syndrome were finally included in the study. All the basic information was gathered and the fresh fecal samples were collected for 16S rDNA sequencing. 16S rDNA of gut microbiota was sequenced using Illumina hiseq 2500 high-throughput sequencing platform.Based on the optimized sequence, OTU clustering analysis and taxonomic annotation were carried out. Results: The difference in relative abundance of gut microbiota was significant between damp heat syndrome and liver depression and spleen deficiency syndrome in CHB patients. Cyanobacteria was only found in damp heat syndrome.The relative abundance of Erysipelotrichia and Subdoligranulum were higher in liver depression and spleen deficiency syndrome,while the relative abundance of Rhodospirillales, Alphaproteobacteria,and Lachnospira were higher in the damp heat syndrome.LEfSe analysis showed that Lachnospira,Olsenella and Subdoligranulum had significant difference in species among the two TCM syndromes.Conclusions: The different characteristics of gut microbiota in the two TCM syndromes of CHB patients may play an important role in syndrome formation of TCM,which provides a new field of vision for the accurate diagnosis and treatment of TCM.

scholarly journals Study on the Characteristics of Gut Microbiota in Chronic Hepatitis B Patients with Damp Heat Syndrome and Liver Depression and Spleen Deficiency Syndrome

2020 ◽  
Author(s):  
Kaiping Jiang ◽  
Qunfang Jiang ◽  
Xiaoai Mo ◽  
Jianhong Li ◽  
Hongtao Hu ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
16S Rdna ◽  
Relative Abundance ◽  
High Throughput Sequencing ◽  
Deficiency Syndrome ◽  
Spleen Deficiency ◽  
Rdna Sequencing ◽  
Significant Difference ◽  
Heat Syndrome ◽  
Spleen Deficiency Syndrome

Abstract Background: To explore the potential biological characteristics of traditional Chinese medicine(TCM) syndromes in CHB patients is of great significance to improve the level of TCM treatment.This study was designed to explore the characteristics of gut microbiota in CHB patients with two most common TCM Syndromes.Methods: According to the diagnostic criteria of Western medicine, TCM and screening exclusion criteria, 65 cases of CHB patients with damp heat syndrome and 28 cases of CHB patients with liver depression and spleen deficiency syndrome were finally included in the study. All the basic information was gathered and the fresh fecal samples were collected for 16S rDNA sequencing. 16S rDNA of gut microbiota was sequenced using Illumina hiseq 2500 high-throughput sequencing platform.Based on the optimized sequence, OTU clustering analysis and taxonomic annotation were carried out. Results: The difference in relative abundance of gut microbiota was significant between damp heat syndrome and liver depression and spleen deficiency syndrome in CHB patients. Cyanobacteria was only found in damp heat syndrome.The relative abundance of Erysipelotrichia and Subdoligranulum were higher in liver depression and spleen deficiency syndrome,while the relative abundance of Rhodospirillales, Alphaproteobacteria,and Lachnospira were higher in the damp heat syndrome.LEfSe analysis showed that Lachnospira,Olsenella and Subdoligranulum had significant difference in species among the two TCM syndromes.Conclusions: The different characteristics of gut microbiota in the two TCM syndromes of CHB patients may play an important role in syndrome formation of TCM,which provides a new field of vision for the accurate diagnosis and treatment of TCM.

Metabolite fingerprinting of novel Streptomyces UK-238 from the Himalayan forest

2020 ◽  
Vol 16 ◽  
Author(s):  
Nidhi Srivastava ◽  
Indira P. Sarethy
Keyword(s):  
Ethyl Acetate ◽  
16S Rdna ◽  
Retention Index ◽  
Ethyl Acetate Extract ◽  
Similarity Index ◽  
Antimicrobial Drug ◽  
16S Rdna Sequencing ◽  
Metabolite Fingerprinting ◽  
Streptomyces Sp ◽  
Rdna Sequencing

Aims: Characterization of antimicrobial metabolites of novel Streptomyces sp. UK-238. Background: Novel antimicrobial drug discovery is urgently needed due to emerging multi antimicrobial drug resistance among pathogens. Since many years, natural products have provided the basic skeletons for many therapeutic compounds including antibiotics. Bioprospection of un/under explored habitats and focussing on selective isolation of actinobacteria as major reservoir of bio and chemodiversity has yielded good results. Objective: The main objectives of the study were the identification of UK-238 by 16S rDNA sequencing and antimicrobial metabolite fingerprinting of culture extracts. Method: In the present study, a promising isolate, UK-238, has been screened for antimicrobial activity and metabolite fingerprinting from the Himalayan Thano Reserve forest. It was identified by 16S rDNA sequencing. Ethyl acetate extract was partially purified by column chromatography. The pooled active fractions were fingerprinted by GC-MS and compounds were tentatively identified by collated data analysis based on Similarity Index, observed Retention Index from Databases and calculated Retention Index. Results: UK-238 was identified as Streptomyces sp. with 98.4% similarity to S. niveiscabiei. It exhibited broad-spectrum antibacterial and antifungal activity. GC-MS analysis of active fractions of ethyl acetate extract showed the presence of eighteen novel antimicrobial compounds belonging to four major categories- alcohols, alkaloid, esters and peptide. Conclusion: The study confirms that bioprospection of underexplored habitats can elaborate novel bio and chemodiversity.

Plant MicroRNAs Responsive to Fungal Infection

2014 ◽  
Vol 941-944 ◽  
pp. 1141-1145 ◽  
Author(s):  
Hui Li Zhang ◽  
Lin Chen ◽  
Wen Na Li ◽  
Li Li Wang ◽  
Hong Yu Xie
Keyword(s):  
Fungal Pathogens ◽  
High Throughput Sequencing ◽  
Adaptive Immune System ◽  
Messenger Rnas ◽  
Disease Mechanism ◽  
Adaptive Immune ◽  
Alternative Approach ◽  
Crop Disease ◽  
Transcriptional Gene Regulation ◽  
And Control

MicroRNAs (miRNAs) are endogenous small RNAs transcribed from non-coding DNA, which have the capacity to base pair with the target mRNAs (messenger RNAs) to repress their translation or resulted in cleavage. We have paid much attention on the DNA and its coded proteins, the discovery of miRNAs as gene negatively regulators has led to a fundamental change in understanding of post-transcriptional gene regulation in plants. Fungal pathogens infection is the main cause of most economic crops diseases. Unlike humans, plants don’t evolved to have a adaptive immune system, they protect themselves with a mechanism consists of activation and response. Recently, high throughput sequencing validated that miRNA play a crucial role in plant-fungus interaction. A better understanding of miRNA-mediated disease mechanism in fungi should clarify the strategy of crop disease control. MiRNA-based manipulations as gene suppressors, such as artificial miRNAs, may emerge as a new alternative approach for the improvement of crops and control of crop disease.

scholarly journals Environmental Sequencing Provides Reasonable Estimates of the Relative Abundance of Specific Picoeukaryotes

2016 ◽  
Vol 82 (15) ◽  
pp. 4757-4766 ◽  
Author(s):  
Caterina R. Giner ◽  
Irene Forn ◽  
Sarah Romac ◽  
Ramiro Logares ◽  
Colomban de Vargas ◽  
...  
Keyword(s):  
Microbial Diversity ◽  
High Throughput ◽  
Relative Abundance ◽  
Ribosomal Dna ◽  
18S Rdna ◽  
High Throughput Sequencing ◽  
454 Sequencing ◽  
Epifluorescence Microscopy ◽  
Environmental Sequencing

ABSTRACTHigh-throughput sequencing (HTS) is revolutionizing environmental surveys of microbial diversity in the three domains of life by providing detailed information on which taxa are present in microbial assemblages. However, it is still unclear how the relative abundance of specific taxa gathered by HTS correlates with cell abundances. Here, we quantified the relative cell abundance of 6 picoeukaryotic taxa in 13 planktonic samples from 6 European coastal sites using epifluorescence microscopy on tyramide signal amplification-fluorescencein situhybridization preparations. These relative abundance values were then compared with HTS data obtained in three separate molecular surveys: 454 sequencing of the V4 region of the 18S ribosomal DNA (rDNA) using DNA and RNA extracts (DNA-V4 and cDNA-V4) and Illumina sequencing of the V9 region (cDNA-V9). The microscopic and molecular signals were generally correlated, indicating that a relative increase in specific 18S rDNA was the result of a large proportion of cells in the given taxa. Despite these positive correlations, the slopes often deviated from 1, precluding a direct translation of sequences to cells. Our data highlighted clear differences depending on the nucleic acid template or the 18S rDNA region targeted. Thus, the molecular signal obtained using cDNA templates was always closer to relative cell abundances, while the V4 and V9 regions gave better results depending on the taxa. Our data support the quantitative use of HTS data but warn about considering it as a direct proxy of cell abundances.IMPORTANCEDirect studies on marine picoeukaryotes by epifluorescence microscopy are problematic due to the lack of morphological features and due to the limited number and poor resolution of specific phylogenetic probes used in fluorescencein situhybridization (FISH) routines. As a consequence, there is an increasing use of molecular methods, including high-throughput sequencing (HTS), to study marine microbial diversity. HTS can provide a detailed picture of the taxa present in a community and can reveal diversity not evident using other methods, but it is still unclear what the meaning of the sequence abundance in a given taxon is. Our aim is to investigate the correspondence between the relative HTS signal and relative cell abundances in selected picoeukaryotic taxa. Environmental sequencing provides reasonable estimates of the relative abundance of specific taxa. Better results are obtained when using RNA extracts as the templates, while the region of 18S ribosomal DNA had different influences depending on the taxa assayed.

scholarly journals Assessing diversity of the female urine microbiota by high throughput sequencing of 16S rDNA amplicons

2011 ◽  
Vol 11 (1) ◽  
pp. 244 ◽  
Author(s):  
Huma Siddiqui ◽  
Alexander J Nederbragt ◽  
Karin Lagesen ◽  
Stig L Jeansson ◽  
Kjetill S Jakobsen
Keyword(s):  
16S Rdna ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Female Urine

scholarly journals 2-Way k-Means as a Model for Microbiome Samples

2017 ◽  
Vol 2017 ◽  
pp. 1-7 ◽  
Author(s):  
Weston J. Jackson ◽  
Ipsita Agarwal ◽  
Itsik Pe’er
Keyword(s):  
Unsupervised Learning ◽  
16S Rdna ◽  
Mixture Model ◽  
Vaginal Microbiome ◽  
16S Rdna Sequencing ◽  
Sequencing Data ◽  
Cluster Assignment ◽  
Rdna Sequencing

Motivation. Microbiome sequencing allows defining clusters of samples with shared composition. However, this paradigm poorly accounts for samples whose composition is a mixture of cluster-characterizing ones and which therefore lie in between them in the cluster space. This paper addresses unsupervised learning of 2-way clusters. It defines a mixture model that allows 2-way cluster assignment and describes a variant of generalized k-means for learning such a model. We demonstrate applicability to microbial 16S rDNA sequencing data from the Human Vaginal Microbiome Project.

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