The stethoscope is a universal tool in the hospital that is in direct contact with many patients and can therefore be a vector in the dissemination of bacterial infections. To research the presence of bacteria, fungi and yeast on the stethoscope diaphragm and the resistance of bacteria to antimicrobial drugs. Descriptive, prospective, non-controlled. A tertiary care hospital. Samples were taken randomly from 300 stethoscopes employed by medical staff (medical residents, medical students, nurses and nursing school students) and other sectors of the hospital. Three hundred stethoscope diaphragms used in several sectors of the hospital facilities by medical doctors (63 samples), medical residents (54 samples), medical students (106 samples), nursing school students (33 samples) and specific sectors (36 samples) were analyzed. Material was collected randomly. It was collected with the aid of a sterile swab moistened in physiological solution, inoculated into Brain Heart Infusion media and incubated in an oven for 24 to 48 hours. After this period, the samples were inoculated into blood agar, MacConkey agar and Sabouraud media and identified by Gram staining and biochemical assays. An assay to test bacteria sensitivity to antibiotics was also carried out by the Kirby-Bauer method. Eighty-seven percent of the analyzed stethoscopes were contaminated. Gram-positive cocci, yeasts, fungi and Gram-positive and negative bacilli were isolated. There was no significant association between the most predominant microorganisms and professional category. Staphylococcus aureus, Staphylococcus negative coagulase and Bacillus were significantly more frequent in relation to the presence of more than one microorganism on the stethoscope diaphragm. Stethoscopes presented a high rate of contamination and their use without precautions can spread nosocomial infections.