Dataset Information

High-Throughput Sequencing and Unsupervised Analysis of Formyltetrahydrofolate Synthetase (FTHFS) Gene Amplicons to Estimate Acetogenic Community Structure.

ABSTRACT: The formyltetrahydrofolate synthetase (FTHFS) gene is a molecular marker of choice to study the diversity of acetogenic communities. However, current analyses are limited due to lack of a high-throughput sequencing approach for FTHFS gene amplicons and a dedicated bioinformatics pipeline for data analysis, including taxonomic annotation and visualization of the sequence data. In the present study, we combined the barcode approach for multiplexed sequencing with unsupervised data analysis to visualize acetogenic community structure. We used samples from a biogas digester to develop proof-of-principle for our combined approach. We successfully generated high-throughput sequence data for the partial FTHFS gene and performed unsupervised data analysis using the novel bioinformatics pipeline "AcetoScan" presented in this study, which resulted in taxonomically annotated OTUs, phylogenetic tree, abundance plots and diversity indices. The results demonstrated that high-throughput sequencing can be used to sequence the FTHFS amplicons from a pool of samples, while the analysis pipeline AcetoScan can be reliably used to process the raw sequence data and visualize acetogenic community structure. The method and analysis pipeline described in this paper can assist in the identification and quantification of known or potentially new acetogens. The AcetoScan pipeline is freely available at https://github.com/abhijeetsingh1704/AcetoScan.

SUBMITTER: Singh A

PROVIDER: S-EPMC7481360 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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High-Throughput Sequencing and Unsupervised Analysis of Formyltetrahydrofolate Synthetase (FTHFS) Gene Amplicons to Estimate Acetogenic Community Structure.

Singh Abhijeet A Nylander Johan A A JAA Schnürer Anna A Bongcam-Rudloff Erik E Müller Bettina B

Frontiers in microbiology 20200827

The formyltetrahydrofolate synthetase (FTHFS) gene is a molecular marker of choice to study the diversity of acetogenic communities. However, current analyses are limited due to lack of a high-throughput sequencing approach for FTHFS gene amplicons and a dedicated bioinformatics pipeline for data analysis, including taxonomic annotation and visualization of the sequence data. In the present study, we combined the barcode approach for multiplexed sequencing with unsupervised data analysis to visu ...[more]

PMID: 32983047

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Project description:IntroductionThe mutton quality of Chinese Tan lambs (Ovis aries) has declined as feeding patterns have shifted from pasturing to pen rationing. While pen-fed can enhance the growth performance of sheep, it falls short in terms of meat quality attributes such as meat color and tenderness. Furthermore, compared to pen-fed, pasture-fed husbandry increases the proportion of oxidative muscle fibers, decreases the proportion of glycolytic muscle fibers, and reduces LDH (lactate dehydrogenase) activity. Mutton quality is affected by fatty acids, and rumen microorganisms play a role in the synthesis of short-chain fatty acids, long-chain fatty acids, and conjugated linoleic acids.MethodsWe used 16S rRNA sequencing to analyze the effects of two feeding patterns on the rumen bacteria of Tan lambs. In a randomized block design with 24 newborn Tan lambs, 12 lambs were fed by ewes in pasture and 12 were fed by pen-fed ewes. At 2 months, the biceps femoris and the longissimus dorsi were analyzed by gas chromatography for intramuscular fat content and fatty acids composition, and DNA in the rumen contents was extracted and used to analyze the structure of the bacterial community.ResultsDifferent feeding patterns had no significant effect on the intramuscular fat content of the biceps femoris and longissimus dorsi of the lambs, but there was a significant effect on fatty acids composition. The fatty acids c18:3n3 and c20:5n3 were significantly higher in the biceps femoris and longissimus dorsi of the pasture group than the pen-ration group. The alpha diversity of rumen bacteria was significantly greater in the pasture group compared to the pen-ration group. The ACE index, Chao1 index, Shannon index, and Simpson index were all notably higher in the pasture group than in the pen-ration group. Utilizing beta diversity analysis to examine the differences in rumen bacteria between the pasture group and pen-ration group, it was observed that the homogeneity of bacteria in the pasture group was lower than that in the pen-ration group. Furthermore, the diversity of rumen bacteria in the pasture group was greater than that in the pen-ration group. Twenty-one phyla were identified in the pasture group, and 14 phyla were identified in the pen-ration group. The dominant phyla in the pasture group were Bacteroidetes and Fibrobacteres; the dominant phyla in the pen-ration group were Proteobacteria and Bacteroidetes. The relative abundance of Proteobacteria was significantly higher in the pen-ration group than in the pasture group (p < 0.01). Diversity at the genus level was also higher in the pasture group, with 176 genera in the pasture group and 113 genera in the pen-ration group. The dominant genera in the pasture group were Prevotella_1, Rikenellaceae_RC9_gut_group, and Bacteroidales_BS11_gut_group_Na; the dominant genera in the pen-ration group were Prevotella_1, Prevotella_7, Succinivibrionaceae_UCG-001, and Succinivibrionaceae_NA.DiscussionThe rumen bacterial community of Tan sheep is significantly influenced by pen-ration and pasture-fed conditions, leading to variations in fatty acid content in the muscle, which in turn affects the flavor and nutritional value of the meat to some extent. Pasture-fed conditions have been shown to enhance the diversity of rumen bacterial community structure in Tan sheep, thereby increasing the nutritional value of their meat.

Dataset Information

High-Throughput Sequencing and Unsupervised Analysis of Formyltetrahydrofolate Synthetase (FTHFS) Gene Amplicons to Estimate Acetogenic Community Structure.

Publications

High-Throughput Sequencing and Unsupervised Analysis of Formyltetrahydrofolate Synthetase (FTHFS) Gene Amplicons to Estimate Acetogenic Community Structure.

Similar Datasets

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