Background: Invasive candidiasis (IC) has emerged worldwide as an important healthcare associated infection caused by Candida species. Nowadays, data on the epidemiology of IC and the antifungal susceptibility of Candida isolates in Tunisia are still limited. Hence, this study aimed to analyze the incidence, species distribution and antifungal susceptibility of Candida strains in the university hospital Sahloul (Sousse, Tunisia). Methods: A five-year (2016 to 2020) retrospective study was conducted including all Candida isolates recovered from blood, other sterile body fluids and deep-seated samples from hospitalized patients. Candida species were identified using the VITEK®2 automated system and the antifungal susceptibilities were determined by the Vitek®2 YST cards and/or the E-test method. Results: During the reported period, 138 nonrepetitive Candida isolated from separate patients were identified with a mean annual incidence of 1,04 case per 1000 admissions. Candida albicans (C. albicans) was the predominant species (42%), followed by C. parapsilosis (19,6%), C. tropicalis (11,6%) and C. glabrata (6,5%). The total resistance rates of fluconazole, voriconazole, amphotericin B and echinocandins were 6,7%, 2,9%, 2,9% and 3,4% respectively. The highest antifungal resistance rates were found with fluconazole (6,7%). This rate significantly increased from 5.3% in 2017 to 10,5% in 2020 (p=0,01). Echinocandins showed excellent in-vitro activities against the majority of Candida species. Conclusion: The present study provides valuable local surveillance data on the epidemiology and antifungal susceptibilities of invasive Candida species which can be useful to guide an appropriate empiric and targeted antifungal therapy.