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44 reviewsSUMMARYAncient genomic studies have extensively explored human-microbial interactions, yet research on nonhuman animals remains limited. In this study, we analyzed ancient microbial DNA from 483 mammoth remains spanning over 1 million years, including 440 newly sequenced and unpublished samples from a1.1-million-year-old steppe mammoth. Using metagenomic screening, contaminant filtering, damagepattern analysis, and phylogenetic inference, we identified 310 microbes associated with differentmammoth tissues. While most microbes were environmental or post-mortem colonizers, we recoveredgenomic evidence of six host-associated microbial clades spanning Actinobacillus, Pasteurella, Streptococcus, and Erysipelothrix. Some of these clades contained putative virulence factors, including aPasteurella-related bacterium that had previously been linked to the deaths of African elephants. Notably,we reconstructed partial genomes of Erysipelothrix from the oldest mammoth sample, representing theoldest authenticated host-associated microbial DNA to date. This work demonstrates the potential ofobtaining ancient animal microbiomes, which can inform further paleoecological and evolutionaryresearch.This is a