Due to the widespread use of second hand clothes recently on a large scale, and the ability of clothing to retain microorganisms, so the research aimed to isolate and characterize microbial contaminants associated with second hand clothes as it is a general concern for everyone due to the possibility of infectious diseases so the purpose of the study was to verify the level and type Bacterial contamination on second hand clothes in order to assess the risks of transmission of pathogenic microorganisms, so the researcher collected samples of different materials and fabrics of second hand clothes . The study used culture methods to isolate pathogenic microbes from second hand clothes. The study concluded that there are statistically significant differences between the average scores of natural fibers, blended fibers, synthetic fibers for each of the textile compositions Plain Weave, Twill Weave and knitted fabrics in (Total count, Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli, yeasts, and fungi counts).