Longitudinal dynamics of the gut microbiome during methamphetamine addiction and abstinence

Methamphetamine (METH) addiction is a chronic neuropsychiatric disorder characterized by severe abstinence symptoms and high relapse rates, yet the temporal dynamics of gut microbiota alterations across addiction and abstinence phases remain poorly understood.

In this study, we performed a longitudinal microbiome analysis to characterize differences between phases and time-dependent remodeling of the gut microbiota in a mouse model. Fecal samples were collected at defined time points across a 60-day addiction phase followed by a 60-day abstinence phase and analyzed using 16S rRNA gene sequencing.

Distinct microbial compositions, diversity and community structure differences were observed between addiction and abstinence phases. Microbial communities during the addiction phase exhibited greater dispersion, whereas progressive stabilization was observed during abstinence. Time-series analyses revealed characteristic alterations in Verrucomicrobia and Actinobacteria during prolonged addiction and early abstinence. At the genus level, Butyricimonas, Candidatus Arthromitus, Enterococcus, and Turicibacter exhibited reproducible phase- and temporal-dependent dynamic patterns. Based on these microbial signatures, random forest models were constructed for phase classification and temporal prediction. The classification model achieved higher model performance during the addiction phase, whereas performance declined during abstinence, reflected by reduced variance explained and increased heterogeneity in gut microbiota composition. Temporal prediction models further demonstrated differential predictive performance between phases.

These findings identify reproducible temporal changes in gut microbiota across addiction and abstinence and highlight candidate microbial signatures for understanding addiction progression and distinguishing different phases.