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Introduction - Clostridium difficile (CD) is the most common cause of health-care-associated infectious diarrhea and incidence and severity have increased in recent years. The main cause of hospital’s acquired cross infections can be attributed to incorrect hand hygiene. We described the epidemiology of CD infection (CDI) in a teaching hospital in Southern Italy during a two years surveillance period and evaluated the health-care workers compliance to hand hygiene. Methods - CDI Incidence rates were calculated as the number of patients with positive C. difficile toxin assay per 10,000 patient-days. Compliance with hand hygiene was the ratio of the number of performed actions to the number of opportunities observed. Approximately 400 Hand Hygiene (HH) opportunities/year /ward were observed. We finally checked out if any correlation could be found. Results - From January 2015 to December 2016 a total number of 854 CD determinations were performed in patients with clinical symptoms of diarrhea. The search for toxins A and B was positive in 175 cases (21,2 %), confirming the diagnosis of CDI. Compliance to hand hygiene was significantly inversely associated with the number of CDIs: the lower the compliance of health-care workers with hand hygiene the higher was the number of cases of CDIs (p=0.003). Conclusion - According to our results proper handwashing of health-care workers appears to be a key intervention in interrupting CD cross infections regardless of age and type of department in which the patient is admitted., Journal of Preventive Medicine and Hygiene, Vol 59, No 2 (2018): 2018592