Microbiomes on surfaces in kindergartens, the intermediate transfer medium for microbial exchange, can exert significant impact on the hygiene and wellbeing of young children, both individually and as a community. Here employing 2bRAD-M, a novel species-resolved metagenomics approach for low-biomass microbiomes, we surveyed over 100 samples from seven frequently contacted surfaces by children, plus individual children’s palms, in two kindergartens. Microbiome compositions, although kindergarten-specific, were grouped closely based on the type of surface within each kindergarten. Extensive microbial admixture were found among the various sampled sites, likely facilitated by contact with children’s hands. Notably, bacterial species with potential human health concerns and potentially antibiotic-resistant, although found across all sampled locations, were predominantly enriched on children’s hands instead of on the environmental sites. This first species-resolved kindergarten microbiome survey underscores the importance of good hand hygiene practices in kindergartens and provides insights into better managing hygiene levels and minimizing spread of harmful microbes in susceptible indoor environments.