Weinstein, I. | bioRxiv Science

bioRxiv2025

Lazypipe3: Customizable Virome Analysis Pipeline Enabling Fast and Sensitive Virus Discovery from NGS data

Metagenomic next-generation sequencing (mNGS) is pivotal for detecting known and novel viruses in diverse samples; however, its efficacy relies on robust bioinformatics pipelines. We present LazypipeX with multiple advanced features, including flexible annotation strategies that can be adjusted to match different datasets and applications. Using a synthetic benchmark, we show that a simple two-round annotation strategy designed for speed and low false positive rate required in virus diagnostics can reduce execution time to a fraction of the BLASTN search without any loss in accuracy. Additionally, using real data, we show that annotation strategies based on combinations of fast-to-accurate homology searches reduce execution time from 5- to 20-fold compared to the BLASTN/BLASTP baseline. Using one of these "annotation chains", we characterized multiple novel complete viral genomes that were missed by the Lazypipe v1/v2 and CZ ID analyses. LazypipeX is a highly efficient and versatile tool for virome analysis, offering customizable and transparent workflows that can facilitate virus discovery and identification in diverse mNGS applications.

bioinformatics↗

bioRxiv2024

Time-series sewage metagenomics can separate the seasonal, human-derived and environmental microbial communities, holding promise for source-attributed surveillance

Sewage metagenomics has risen to prominence in urban population surveillance of pathogens and antimicrobial resistance (AMR). Unknown species with similarity to known genomes cause database bias in reference-based metagenomics. To improve surveillance, we designed this study to recover sewage genomes and develop a quantification and correlation workflow for these genomes and AMR over time. We used longitudinal sewage sampling in seven treatment plants from five major European cities to explore the utility of catch-all sequencing of these population-level samples. Using metagenomic assembly methods, we recovered 2,332 metagenome-assembled genomes (MAGs) from prokaryotic species, 1,334 of which were previously undescribed. These genomes account for [~]69% of sequenced DNA and provide insight into sewage microbial dynamics. Rotterdam (Netherlands) and Copenhagen (Denmark) showed strong seasonal microbial community shifts, while Bologna, Rome, (Italy) and Budapest (Hungary) had occasional blooms of Pseudomonas-dominated communities, accounting for up to [~]95% of sample DNA. Seasonal shifts and blooms present challenges for effective sewage surveillance. We find that bacteria of known shared origin, like human gut microbiota, form communities, suggesting the potential for source-attributing novel species and their ARGs through network community analysis. This could significantly improve AMR tracking in urban environments.

microbiology↗