Here, a simple processing route is introduced for preparation of N-doped nano structure ZnO at 75-80°C using in-situ sonosynthesis method through hydrolysis of zinc acetate at pH≈9-10 adjusting with ammonia. Synthesis and fabrication of nano N-doped ZnO were carried out on the wool fabric through impregnation of the fabric in ultrasound bath using different concentrations of zinc acetate followed by curing. The antibacterial and antifungal activities of the treated fabrics were assessed against two common pathogenic bacteria including Escherichia coli, Staphylococcus aureus and the diploid fungus namely Candida albicans. The photo-catalytic activity of nano N-doped ZnO particles on the wool fabric was determined by degradation of Methylene Blue under daylight irradiation. Increasing zinc acetate and prolonged sonication time led to higher photo-catalytic activity as more dye stain degraded from the stained treated fabric under daylight. Higher photo-catalytic activity was observed on the nano N-doped ZnO sonotreated wool fabric having more hydrophilicity. Finally, the treatment indicated no negative effect on the fabric safety while reduced alkaline solubility and yellowness even enhanced the fabric tensile strength. The response surface methodology was also utilized to optimize the wool fabric treatment conditions.