bioRxiv Science⌕ Search

bioRxiv · 10.1101/2025.06.30.662361

Stool Dynamics and the Developing Gut Microbiome During Infancy

Abstract

The infant gut microbiome is a dynamic ecosystem that plays a crucial role in early development, influences immune system maturation, and overall health. Recent insights reveal that the gut microbiota undergoes changes across the 24-h day, raising the possibility that it may act as "zeitgeber", helping to regulate the hosts sleep-wake organisation. Despite its importance, timing factors influencing microbiome composition are poorly understood, limiting its use as a health predictor. This study investigates the relationship between stool dynamics (interval since last defecation, collection time of the sample), sleep pressure (interval since last sleep), meal timing and the composition of the gut microbiome. Stool samples from 198 healthy infants, aged 3 to 31 months, were analysed to assess microbial diversity, richness evenness, and abundance. Our findings reveal that longer intervals since the last bowel movement are associated with increased microbial diversity, evenness, and richness. Stool timing is associated with shifts in microbial composition, especially in younger infants, indicating that diurnal microbial fluctuations may become more stable as infants mature. We found that longer periods of wakefulness were associated with increased microbial diversity in early infancy, although this effect appeared to diminish with age. Feeding schedules appeared to have a limited effect on the gut microbiome. Longer fasting before sampling showed no significant associations with most microbial parameters, except for a positive association with microbial richness. At the phylum level, results indicate that infant gut microbial composition is influenced by behaviour and physiology. Longer intervals between bowel movements were associated with shifts in bacterial abundance, with Proteobacteria decreasing and Actinobacteria increasing. Stool timing and meal timing also affected microbial diversity: later sampling times showed higher Actinobacteria levels, and longer fasting was associated with reduced Bacteroidetes. Sleep pressure showed a trend effect with Firmicutes displaying a slight decrease in infants who had been awake longer. Our findings underscore the importance of time-based factors on infant gut microbiome composition.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Al-Andoli, M., Schoch, S., Markovic, A., Muhlematter, C., Beaugrand, M., Jenni, O., Liamlahi, R., Walser, J.-C., Nielsen, D. S., Kurth, S.. 2025-07-04. Stool Dynamics and the Developing Gut Microbiome During Infancy. https://doi.org/10.1101/2025.06.30.662361

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Concentration limits and localization of hydrogen peroxide in the extracellular space of solid tissues

H2O2 released to the extracellular space (ECS) regulates diverse physiological processes, yet its concentrations and spatial distribution in tissues remain poorly defined. This uncertainty hampers mechanistic understanding of redox signaling. Here, we used reaction-diffusion modeling to estimate extracellular H2O2 concentrations and transport ranges in various scenarios. Idealized analytical models were combined with numerical models incorporating localized NADPH oxidase (NOX) clusters, ECS microstructure, membrane permeability, and the thioredoxin- and GSH-dependent clearance systems. Using maximal neutrophil and NOX superoxide/H2O2 release rates, we obtained upper bounds for extracellular H2O2. Adjacent to isolated average-sized, fully active NOX2 clusters H2O2 peaked at ~540 nM at adhesion cell-cell separations, and decreased radially over ~50-100 nm. At the receptor cell surface, peak concentration decreased inversely with intercellular separation, to <5 nM at 1 m separation. Radial decrease here, for this wide separation, was over ~2.5 m. Even the former maximal extracellular concentrations induce just a minimal, highly localized oxidation of the intracellular Prdx, Trx and GSH pools. In turn, maximally activated neutrophils carry ~2000 such NOX2 clusters, inducing 10s of M peak ECS H2O2 concentrations. These cause extensive Prdx and Trx oxidation near the exposed membranes. However, the GSH-dependent system still sustains a strong transmembrane gradient if the permeation barrier remains intact, and ECS H2O2 concentrations decay to sub-M within a few m of the source cell. Extracellular H2O2 concentrations scaled linearly with source flux in all the examined conditions. These results establish stringent constraints on autocrine, juxtacrine and next-cell paracrine H2O2 signaling.

systems biology↗

LSD-pipeline: Causal Inference of miRNA Network Effects in Alzheimer's Disease

MicroRNAs (miRNAs) are implicated in Alzheimer's disease (AD), but research has focused on individual miRNAs and direct targets. Existing approaches to miRNA regulation in AD identify associations rather than causal effects, and few methods estimate multi-stage chains from miRNAs through target genes to target transcription factor (TF) cascades. We developed the LSD pipeline (LASSO-SEM-DoWhy), integrating LASSO feature selection, multi-stage structural equation modeling, and DoWhy causal inference to identify and validate miRNA causal pathways in AD. Applying LSD to six blood miRNA and brain mRNA datasets, we identified four LSD-validated miRNAs (miR-30d-5p, miR-92a-3p, miR-296-5p, miR-193a-5p) as AD biomarkers, achieving >86% ROC accuracy in an independent validation cohort. Several miRNAs with no significant direct association with AD showed significant effects when estimated through their target networks, while others significant in direct analysis were not supported at the network level, underscoring the value of network-level analysis. Extending to the TF layer revealed complete miRNA [->] targets [->] TF cascades [->] AD causal chains, with HMGA1, NKX2-3, and PRRX2 as key intermediaries. Confirmed classic pathways converge primarily on tau pathology and synaptic dysfunction. miRNA effects were largely age-independent, suggesting miRNAs act as early initiators of AD pathogenesis. Beyond AD, the LSD pipeline provides a generalizable framework for uncovering causal regulatory mechanisms in other diseases.

systems biology↗

PyKappa: Rule-based modeling in Python

Rule-based languages have proven effective for modeling systems of interacting structured entities as typically encountered in chemistry and molecular biology. We present PyKappa, a rule-based modeling package written in Python whose interpreted nature enables interactive simulation and analysis, including by agentic AI. The package seeks to broaden the base of developers by utilizing a widely known programming language and serves as an easy-to-deploy teaching tool. Using PyKappa, we conduct a case study of phase separation.

systems biology↗