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Growing demand for sustainable and green technologies has turned industries and research toward the more efficient utilization of natural and renewable resources. In an effort to tackle this issue, we developed an antibacterial textile nanocomposite material based on cotton and peat fibers with immobilized Cu-based nanostructures. In order to overcome poor wettability and affinity for Cu²⁺-ions, the substrate was activated by corona discharge and coated with the biopolymer chitosan before the in situ synthesis of nanostructures. Field emission scanning electron microscopy (FESEM) images show that the application of gallic or ascorbic acid as green reducing agents resulted in the formation of Cu-based nanosheets and mostly spherical nanoparticles, respectively. X-ray photoelectron spectroscopy (XPS) analysis revealed that the formed nanostructures consisted of Cu₂O and CuO. A higher-concentration precursor solution led to higher copper content in the nanocomposites, independent of the reducing agent and chitosan deacetylation degree. Most of the synthesized nanocomposites provided maximum reduction of the bacteria <i>Escherichia coli</i> and <i>Staphylococcus aureus</i>. A combined modification using chitosan with a higher deacetylation degree, a 1 mM solution of CuSO₄ solution, and gallic acid resulted in an optimal textile nanocomposite with strong antibacterial activity and moderate Cu²⁺-ion release in physiological solutions. Finally, the Cu-based nanostructures partially suppressed the biodegradation of the textile nanocomposite in soil.