Introduction: Enterococcus sp is a bacterium that is part of the intestinal microbiota and contaminates the female genital tract. Generally, it does not cause problems; however, it can develop infection if it enters the urinary tract, bloodstream, skin wounds, or other sterile areas. Objective: To determine and quantify the presence of Enterococcus sp on inert surfaces of restrooms in the Academic Department of Health and Wellness of the Catholic University of Cuenca. Methodology: A non-experimental, quantitative cross-sectional study was conducted; 424 samples from 106 restrooms were collected. The samples were gathered from door and toilet flush handles in the different restrooms of the Academic Department of Health and Wellness employing the swab technique and subsequent isolation in chromogenic culture medium. Then, the samples were taken to the Microbiology Laboratory of the Academic Department of Biochemistry and Pharmacy of the Catholic University of Cuenca. Results: A higher presence of Enterococcus sp was determined in the restrooms before routine cleaning, representing 1.44% of the total, while after cleaning, it was 0.48%. Conclusion: The study revealed that restrooms have a higher bacterial load of Enterococcus sp before bathroom cleaning.