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The first evidence of the atrial natriuretic peptide (ANP) effect on mono-species and dual-species biofilms of skin commensals <i>Cutibacterium acnes</i> and <i>Staphylococcus epidermidis</i> was obtained in different model systems. Elucidation of the mechanism of action of hormones on the microbial communities of human skin is an important physiological and medical aspect. Under anaerobic conditions, ANP at a concentration of 6.5 × 10⁻¹⁰ M inhibits the growth of <i>S. epidermidis</i> biofilms and stimulates the growth of <i>C. acnes</i> biofilms, and a lesser effect has been demonstrated on planktonic cultures. In biofilms, ANP stimulates aggregation in <i>C. acnes</i> and aggregate dispersion of <i>S. epidermidis</i>, while in <i>S. epidermidis</i>, ANP also stimulates the metabolic activity of cells. Analysis of dual-species biofilms has shown the dominance of <i>S. epidermidis</i>, while ANP increases the ratio of <i>C. acnes</i> biomass in the community. ANP decreases the growth rate of <i>S. epidermidis</i> biofilms and increases that of <i>C. acnes</i>. The effect of ANP is not dependent on the surface type and probably affects other targets in microbial cells. Thus, the potential regulatory effect of human ANP on skin microbe dual-species communities has been shown, and its potential has been demonstrated to change microbiota homeostasis on the skin.