Antimicrobial treatments have become more important for the textile materials especially used in sportswear, activewear, and casual wear since they can easily be contaminated by perspiration leading to bacterial growth and body odor. In this work, antimicrobial activity of chitosan in a silica matrix on pretreated wool fabrics was studied. The pretreatment processes were applied by two different ways (enzymatic and enzymatic+hydrogen peroxide). Afterwards chitosan solutions were applied to the untreated samples and to the samples that were pretreated by two different ways to give antimicrobial effects. The antimicrobial activity of wool fabrics treated in various methods was assessed before and after repeated washings (up to 10 cycles) by the application of standard test method AATCC 147-1998. The morphology of the treated fabrics was investigated by SEM and their characterizations were made by the FT-IR spectral analysis. Results revealed that pretreatment ways and chitosan application methods were quite important for adsorption and diffusion of chitosan on wool fabrics and washing stability. From the SEM images, it was clearly observed that pretreatment processes caused some degradation on the surface of the fiber; but combined processes were found to be less degradative and more effective.