Current evidence of hotel room cleanliness is based on observation rather than empirically based microbial assessment. The purpose of the study described here was to determine if observation provides an accurate indicator of cleanliness. Results demonstrated that visual assessment did not accurately predict microbial contamination. Although testing standards have not yet been established for hotel rooms and will be evaluated in Part II of the authors’ study, potential microbial hazards included the sponge and mop (housekeeping cart), toilet, bathroom floor, bathroom sink, and light switch. Hotel managers should increase cleaning in key areas to reduce guest exposure to harmful bacteria.