The issue of nosocomial infections, or healthcare-associated infections (HAIs), remains a significant concern in healthcare settings worldwide. In recent times, there has been growing attention towards medical devices, notably stethoscopes, as potential vectors for pathogen transmission. This study aimed to evaluate the prevalence of Staphylococcal contamination on stethoscopes used by students and staff at Istanbul University-Cerrahpasa Faculty of Veterinary Medicine’s animal hospital. Furthermore, it gathered information about stethoscope usage habits, cleaning practices, handwashing routines, participants' knowledge about nosocomial infections, and their interest in learning more about these infections and stethoscope hygiene. The analysis of 50 stethoscope samples revealed that 27 (54%) were contaminated with one or more Staphylococci. The isolated 30 Staphylococcus spp. included Staphylococcus epidermidis (n=17; 56.7%), Staphylococcus hominis (n=10; 33.3%), Staphylococcus pasteuri (n=1; 3.3%), Staphylococcus capitis (n=1; 3.3%), and Staphylococcus schleiferi (n=1; 3.3%). Notably, the absence of the highly pathogenic Staphylococcus aureus in all samples provides some reassurance. However, the presence of various Staphylococcus spp. raises concerns due to their pathogenic potential. These findings align with previous research on stethoscope contamination, emphasizing the persistent problem of bacterial colonization on these crucial medical devices. Despite variations in bacterial prevalence among studies, Staphylococcus spp. consistently emerge as common contaminants, emphasizing the need for comprehensive stethoscope hygiene protocols in veterinary healthcare settings. To the best of our knowledge, this is the first report describing the vector potential of stethoscopes in a veterinary setting within Turkey. The study suggests the necessity for further research, taking a proactive approach to tackle the challenges of nosocomial infections. This would enable the development of strategies to ensure a safer healthcare environment for patients and healthcare providers.