The year 2020 was characterized by the outbreak of a new pandemic caused by a novel coronavirus named SARS-CoV-2. To face the pandemic, many countries worldwide imposed general lockdowns, closing all non-essential businesses. As primary care services, pharmacies had to remain open, thus putting pharmacy staff at significant risk of viral infection and overwork. This study aimed to assess the mental health of Italian Pharmacists, considering demographic and occupational characteristics, lifestyle, and habits, during the SARS-CoV-2 outbreak and the subsequent lockdown period (March-May 2020). A web-based survey was created using Google® Forms to collect data from March 30, 2020, to June 1, 2020. The questionnaire consisted of three sections investigating: (1) demographic and occupational variables, (2) lifestyle and habits variables, (3) psychological distress and perceived well-being. A total of 401 participants completed the questionnaire. Older workers and those with more work experience reported more psychological stress. Older and female workers, who felt lonely at home and reported psychological stress, perceived poor well-being. Our findings demonstrate that the Sars-CoV-2 outbreak and subsequent lockdown rules affected pharmacists' mental health and that it is important to put in place preventive measures against the occurrence of mental disorders among them.