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Background and Objectives: Microbiological contamination of sanitary examination gloves and its dispensing method has been poorly studied. Our objectives were to find out in which dispensing model the gloves are most contaminated, to quantify colony-forming units (CFU), to assess whether services influence contamination, and to identify the microorganisms involved. Material and Methods: Transversal descriptive study. Two dispensing models were studied in the emergency department, examination rooms, ICU, the post anesthesia resuscitation unit, and isolated patient rooms: the conventional dispensing (horizontal placement of the box, multiple gloves extraction and non-selective manipulation when extracting), and the alternative dispensing (vertical placement of the box, individual extraction and selective manipulation when extracting). Samples were taken on chocolate agar plates. Results: 92 gloves were analyzed. 71.7% of the gloves from the conventional model (n=33) were contaminated versus 30.4% of the gloves from the alternative model (n=14), with a prevalence ratio of 2.4 (p<0.001). The contamination median in conventional gloves was 3 CFU (interquartile range of 6.25), while in the alternative model, the median was 0 CFU (interquartile range of 1, p<0.001). There were no statistically significant differences depending on the services. The microorganisms involved in the contamination were coagulase-negative Staphylococcus, Bacillus circulans, and Streptococcus viridans. Conclusions: The design of the glove dispensing boxes influences their contamination. Gloves dispensed in conventional boxes are more contaminated and with a greater amount of CFU than in alternative dispensers. The services do not influence contamination and the microorganisms involved are skin and environmental flora. [ABSTRACT FROM AUTHOR]