Fig. 1: Study design, sample information and analysis by time point. | Nature Communications

Fig. 1: Study design, sample information and analysis by time point.

From: Effects of iron supplements and iron-containing micronutrient powders on the gut microbiome in Bangladeshi infants: a randomized controlled trial

Fig. 1

A BRISC trial schema showing assessment and sampling time points. B Flow diagram outlining stool samples for 16S rRNA and shotgun metagenomic sequencing. C Stacked bar plot presenting taxonomic composition of baseline samples at study entry (i.e., baseline) based on relative abundance of the top five genera overall. Each bar represents a sample, with relative abundance on the y-axis. Samples are grouped by trial arm (16S rRNA data). Taxonomic boxplots showing relative abundance of the four most abundant bacterial phyla by sampling time point (D) 16S rRNA data and (E). shotgun metagenomic data. Boxplot center lines denote median value, with bounds of box indicating 25–75th percentiles. Whisker lines encompass 1.5 x interquartile range from above the upper and below the lower quartiles. Data points outside whiskers are outliers. (n = 900 at baseline, n = 790 at post-intervention and n = 574 at post-follow-up time points for (D), and n = 316 at baseline, n = 316 at post-intervention and n = 312 at post-follow-up time points for E). F Heatmap showing relative differences in abundance of genes relating to microbiome functional profile by sampling time point across all trial arms, with baseline as reference. Relative abundance data underwent centered log ratio normalization/transformation in a general linear model analysis in MaAsLin2, with data for differentially abundant functional pathways including coefficient (approximating log2-fold change), standard deviation and FDR-adjusted p value. Only the 10 features exhibiting the greatest relative change in abundance (increase – in red – or decrease – in blue – from baseline) are shown. Abundance is expressed as −log10(FDR-adjusted p value) * sign(log2-fold change) as per the heatmap calculation used by MaAsLin2. (n = 255 at baseline, n = 231 at post-intervention and n = 312 at post-follow-up time points) (Shotgun metagenomic data). A, B created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en). Source data for Fig. 1C–F are provided in the Source Data file.

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