Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1012831 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:Purpose: This study aims to compare and analyze the differences in bacterial community composition in fecal samples from mice treated with Control(DW), Vancomycin (VAN), Ampicillin (AMP), Neomycin (NEO), Metronidazole (MET), and a combination of all antibiotics (ALL, VANM) using 16S rRNA sequencing. Methods: Each antibiotics treated mice's fecal samples were collected and stored -80'c until analyzation. DNA was extracted using the NucleoSpin DNA Stool Kit (MACHEREY-NAGEL) following the manufacturer’s protocol. Metagenomic sequencing was performed on an Illumina MiSeq platform (Illumina), targeting the V3 and V4 regions of the 16S rRNA gene according to the manufacturer's instructions. PCR products were purified using AMPure XP beads, and sequencing adapters were added using the Nextera XT Index Kit (Illumina). The library was further purified with AMPure XP beads and quantified using automated electrophoresis with the TapeStation System (Agilent). Sequencing was performed using the MiSeq v3 reagent kit (Illumina), following the manufacturer’s protocol. Results: QIIME2 (v2023.02) was used to process and analyze 16S rRNA gene amplicon sequencing data, from sequence preprocessing to taxonomic classification. Paired-end sequences were merged and quality-filtered using Deblur. The resulting amplicon sequence variants (ASVs) were used for downstream analyses. Conclusions: Our study presents a comparative analysis of bacterial community composition in fecal samples from antibiotic-treated mice. We observed that microbiota composition varied distinctly depending on the type of antibiotic administered.
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Project description:The objective of this study was to determine if Salmonella colonization of chickens could be reduced through competitive exclusion using a defined community of chicken commensal bacteria. One-day old White Leghorn chicks, hatched on-site, were randomly divided into experimental groups and given an oral gavage of either a defined community of 15 bacterial species (DC), cecal contents (CC), or sterile PBS (control; CT). After one week, birds were euthanized for cecal content collection (pre-Salmonella sample) while the remaining birds were orally gavaged 1 X 10^8 colony forming units (CFU) of Salmonella enterica ser. Heidelberg strain 2813 (SH2813). Bacterial counts for three post-Salmonella timepoints (3, 14, and 28 days post inoculation; dpi) were evaluated. Bacteriological enumeration was performed by plating cecal contents onto Salmonella selective agar to determine CFU/g in each group for all collection days. Cecal contents were also used for 16S amplicon sequencing. Cecal tissue was used for stranded mRNA sequencing (RNA-Seq).
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Project description:In this study we developed metaproteomics based methods for quantifying taxonomic composition of microbiomes (microbial communities). We also compared metaproteomics based quantification to other quantification methods, namely metagenomics and 16S rRNA gene amplicon sequencing. The metagenomic and 16S rRNA data can be found in the European Nucleotide Archive (Study number: PRJEB19901). For the method development and comparison of the methods we analyzed three types of mock communities with all three methods. The communities contain between 28 to 32 species and strains of bacteria, archaea, eukaryotes and bacteriophage. For each community type 4 biological replicate communities were generated. All four replicates were analyzed by 16S rRNA sequencing and metaproteomics. Three replicates of each community type were analyzed with metagenomics. The "C" type communities have same cell/phage particle number for all community members (C1 to C4). The "P" type communities have the same protein content for all community members (P1 to P4). The "U" (UNEVEN) type communities cover a large range of protein amounts and cell numbers (U1 to U4). We also generated proteomic data for four pure cultures to test the specificity of the protein inference method. This data is also included in this submission.
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