Natural organic fibers such as cotton, linen, and wool are readily attacked by microorganisms. Microbial growth on a textile fabric causes loss of strength and elongation, discoloration, and changes in appearance. The antibacterial finishing agents extracted from nettle plant leaf have been used to impart finish to the cotton fabric by using the Pad-Dry-Cure application method. The antibacterial activity of the finish was assessed quantitatively using the AATCC 100:2004 test method, and the antibacterial activity against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria was measured. There was a 100 to 99.75 percent reduction in the count of test bacteria. Physical properties of treated and untreated cotton fabrics such as absorbency, fabric stiffness, air permeability, and strength were analyzed. To evaluate the laundering effect of stinging nettle leaf finished fabric, samples were subjected to laundry with 5, 10, 15, 20, 25, 30, 35, and 40 washing cycles using standard AATCC test method 61, and bacterial count of treated samples was tested. The results showed gradual decrease in antibacterial property, with a 100 to 44% reduction in the bacterial count for Staphylococcus aureus and a 100 to 30% reduction in the bacterial count for Escherichia coli. The results of this investigation suggested that nettle leaf, a low-cost abundant plant in Ethiopia, can be used for antibacterial activity in woven cotton fabric.