Healthcare-associated infections are still a worrying health problem that complicates patient care and increases morbidity and mortality. The incidence of infections in hemodialysis patients is still high and they constitute the second cause of hospitalization and death in this category. The study's objective was to evaluate nursing and medical staff's involvement in hospital hygiene. This is an observational study that took place within the hemodialysis unit of a Moroccan university hospital from March 2021 to June 2021. A surface swabbing technique for bacteriological examination was also carried out. The study revealed some shortcomings regarding hand hygiene which was respected by the nurses in only 17.5% of sessions and in 42.2% by the doctors. Other shortcomings in asepsis measures, were also noted such as the non-change of gloves after skin preparation (1.03%) and before manual compression. The serological status of patients concerning HBV, HVC, and HIV was also not always known (31.1%). The bacteriological samples from the surfaces also showed colonization of the medical devices and the ultrasound scanner used for the ultrasound-guided introduction of the central catheters. The results of the study showed some deviations concerning the recommendations; however, they are comparable to other studies conducted internationally, especially in terms of hand hygiene. Our study results highlight some shortcomings adherence to hygiene measures like inconsistent disinfection of medical devices and dialysis stations, as well as insufficient hand hygiene practices among some staff members. However, we observed an improvement in practices following the implementation of awareness-raising sessions. To maintain improved hygiene practices and prevent infections, it is crucial to provide ongoing training for staff, ensure adequate resources, and regularly monitor compliance with hygiene standards.