Abstract

MotivationEstimating microbial association networks from high-throughput sequencing data is a common exploratory data analysis approach aiming at understanding the complex interplay of microbial communities in their natural habitat. Statistical network estimation workflows comprise several analysis steps, including methods for zero handling, data normalization and computing microbial associations. Since microbial interactions are likely to change between conditions, e.g. between healthy individuals and patients, identifying network differences between groups is often an integral secondary analysis step. Thus far, however, no unifying computational tool is available that facilitates the whole analysis workflow of constructing, analysing and comparing microbial association networks from high-throughput sequencing data.ResultsHere, we introduce NetCoMi (Network Construction and comparison for Microbiome data), an R package that integrates existing methods for each analysis step in a single reproducible computational workflow. The package offers functionality for constructing and analysing single microbial association networks as well as quantifying network differences. This enables insights into whether single taxa, groups of taxa or the overall network structure change between groups. NetCoMi also contains functionality for constructing differential networks, thus allowing to assess whether single pairs of taxa are differentially associated between two groups. Furthermore, NetCoMi facilitates the construction and analysis of dissimilarity networks of microbiome samples, enabling a high-level graphical summary of the heterogeneity of an entire microbiome sample collection. We illustrate NetCoMi’s wide applicability using data sets from the GABRIELA study to compare microbial associations in settled dust from children’s rooms between samples from two study centers (Ulm and Munich).AvailabilityR scripts used for producing the examples shown in this manuscript are provided as supplementary data. The NetCoMi package, together with a tutorial, is available at https://github.com/stefpeschel/NetCoMi.ContactTel:+49 89 3187 43258; stefanie.peschel@mail.deSupplementary informationSupplementary data are available at Briefings in Bioinformatics online.

Highlights

  • The rapid development of high-throughput sequencing techniques [1,2,3,4] offers new possibilities for investigating the microbiome across different habitats and provides the opportunity to discover relationships between the composition of microbial communities and their environment

  • The results reveal strong differences between the networks based on different association measures as well as different normalization methods whereas the zero replacement methods available in NetCoMi lead to quite similar networks, if the remaining arguments are fixed

  • With NetCoMi we offer an easy-to-use and versatile, integrative R package for the construction, analysis and comparison of microbial networks derived from MGS data

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Summary

Introduction

The rapid development of high-throughput sequencing techniques [1,2,3,4] offers new possibilities for investigating the microbiome across different habitats and provides the opportunity to discover relationships between the composition of microbial communities and their environment. NetCoMi provides a wide range of existing methods for data normalization, zero handling, edge filtering, and a selection of association measures, which can be combined in a modular fashion to generate the microbial networks. Log-ratios have been proposed by Aitchison [45] as the basis for statistical analyses of compositional data as they are independent of the total sum of counts m. Further information on these measures is available in Supplementary material 1. To ensure wide applicability of NetCoMi, the package comprises traditional association measures, which are not suitable for application on read count data in their original form.

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