Abstract
Abstract Introduction. Application of next-generation sequencing technology generated a massive amount of information on the gut microbiome composition used to understand its role in the healthy state and in various diseases. We aimed to provide information on the gut microbiota composition of Romanian subjects diagnosed with ankylosing spondylitis, an immune-mediated arthropathy linked to a genetic predisposition and gut dysbiosis. Methods. Stool samples collected from 25 patients with ankylosing spondylitis and 16 healthy controls were investigated using high-throughput DNA sequencing of 16S rRNA amplicons from seven different hypervariable regions and Ion Torrent PGM instrument. Microbial composition of metagenomic data was analyzed with QIIME software and differential abundance analysis of taxa encompassed linear discriminant analysis effect size (LEfSe). Results. Overall, 14 phyla, 114 families, 114 genera, and 275 species were identified across the 41 samples, the aggregated data revealing as most abundant the phyla Bacteroidetes, Firmicutes, and Proteobacteria, the families Bacteroidaceae, Prevotellaceae, and Ruminococcaceae, the genera Bacteroides, Prevotella, and Faecalibacterium, and Prevotella copri species. Using various cutoffs for abundance and prevalence, core taxonomic members were identified which in general were shared between the patients and controls. However, evidence was gained that the diversity in the microbiomes from the former cohort was lower than for controls and that certain taxa had significantly different abundance between the two groups. Conclusion. This study allowed an informative high-throughput 16S rRNA profiling of the gut microbiota needed to identify microbiome signatures of risk in the autochthonous population with AS.
Highlights
Ankylosing spondylitis (AS) is a chronic progressive inflammatory rheumatic disease which mainly affects the spine and sacroiliac joints and has as dominant clinical features back pain and progressive stiffness of the spine [1]
Reads from libraries generated from the Ion 16STM Metagenomics Kit were assigned Operational Taxonomic Units (OTUs) allowing identification of 14 phyla, 114 families, 114 genera, and 275 species
A significantly lower number of taxa were identified among the AS microbiomes by comparison with the HC microbiomes (p< 0.05), an observation reflected by the alpha diversity measures output by Quantitative Insights into Microbial Ecology (QIIME) showing a lower gut microbiome richness (p
Summary
Ankylosing spondylitis (AS) is a chronic progressive inflammatory rheumatic disease which mainly affects the spine and sacroiliac joints and has as dominant clinical features back pain and progressive stiffness of the spine [1]. Progress has been made in understanding the etiology and pathogenesis of this handicapping disease there are still unclear aspects that need ongoing investigation. Gut microbiota has been traditionally analyzed by culture methods which in spite of improving in time in terms of techniques for cultivation of anaerobic species provided a limited resolution of the analysis. The further application of next-generation sequencing (NGS) technology generated a massive amount of information on the gut microbiome configuration used to progress in understanding the complexity of the microbial communities and their function in health and disease. In order to allow meaningful comparisons of autochthonous data with those accumulated for individuals from other regions, this report provides baseline microbiological information on the gut microbiota composition of Romanian subjects with AS diagnosis as determined through the culture-independent amplification of 16S ribosomal RNA (rRNA) gene and an NGS approach
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