User-friendly computational tools for 16S ribosomal RNA (rRNA) sequencing analysis enable researchers who are not bioinformaticians to analyze and interpret sequencing data from microbial communities. These tools' easy-to-use interfaces belie the sophisticated and rapidly-evolving science of their underlying algorithms. When analyzing 16S data from a simple microbiome experiment, we found that superficially unimportant decisions about the bioinformatic pipeline led to results with radically different biological interpretations. We share these results as a cautionary tale whose moral is that, in 16S analysis, the devil is in the details. Wet bench researchers should therefore strongly consider partnering with bioinformaticians or computational biologists when analyzing 16S data.
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http://dx.doi.org/10.1080/19490976.2020.1747336 | DOI Listing |
Obes Surg
January 2025
Coimbra Institute for Clinical and Biomedical Research (iCBR) Area of Environment, Genetics and Oncobiology (CIMAGO), Faculty of Medicine, University of Coimbra, Coimbra, Portugal.
Background: Bariatric surgery is the most long-term effective treatment option for severe obesity. The role of gut microbiome (GM) in either the development of obesity or in response to obesity management strategies has been a matter of debate. This study aims to compare the impact of two of the most popular procedures, sleeve gastrectomy (SG) and Roux-en-Y gastric bypass (GB), on metabolic syndrome parameters and gut bacterial microbiome and in systemic immuno-inflammatory response.
View Article and Find Full Text PDFSci Rep
January 2025
Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, P.O. Box 2455, 11451, Riyadh, Saudi Arabia.
Polycyclic aromatic compounds (PACs) are pervasive environmental contaminants derived from diverse sources including pyrogenic (e.g., combustion processes), petrogenic (e.
View Article and Find Full Text PDFAnal Chem
January 2025
Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, Indiana 47907, United States.
Propidium monoazide (PMA) is a dye that distinguishes between live and dead cells in molecular assays like the Polymerase Chain Reaction (PCR). It works by cross-linking to the DNA of cells that have compromised membranes or extracellular DNA upon photoactivation, making the DNA inaccessible for amplification. Currently, PMA is used to detect viable pathogens and alleviate systemic bias in the microbiome analysis of samples using 16S rRNA gene sequencing.
View Article and Find Full Text PDFClin Microbiol Infect
January 2025
Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain; Medicine Department, School of Medicine, Universidad Complutense de Madrid (UCM), Madrid, Spain. Electronic address:
Objectives: Faecal microbiota transplantation (FMT) is an established treatment for recurrent Clostridioides difficile infection (R-CDI). This study aimed to identify calprotectin and microbiome characteristics as potential biomarkers of FMT success.
Methods: We conducted a prospective study of patients who underwent oral FMT (single dose of 4-5 capsules) for R-CDI (January 2018 to December 2022).
Environ Res
January 2025
Key laboratory of Exploration and Utilization of Aquatic Genetic Resources of the Ministry of Education, Engineering Research Center of Environmental DNA and Ecological Water Health Assessment, Shanghai Ocean University, Shanghai 201306, China. Electronic address:
Global change stressors, including climate warming, eutrophication, and small-sized omnivorous fish, may exert interactive effects on the food webs and functioning of shallow lakes. Periphyton plays a central role in the primary production and nutrient cycling of shallow lakes but constitutes a complex community composed of eukaryotes and prokaryotes that may exhibit different responses to multiple environmental stressors with implications for the projections of the effects of global change on shallow lakes. We analyzed the effects of warming, nutrient enrichment, small omnivorous fish and their interactions on eukaryotic and prokaryotic periphyton structures in shallow lake mesocosms.
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