In Tunisia, hepatitis A virus (HAV) represents a public health concern. Due to the progress in sanitation and socio-economic conditions, the epidemiology of HAV has shown dynamic changes over the past years. This study aimed to investigate the current seroprevalence of HAV antibodies (anti-HAV) among the residents of Thala, a rural setting in central-west Tunisia, to determine the age specific seroprevalence for HAV infection and co-infection with hepatitis C and B virus. A total of 1379 subjects (mean age: 25.0 ± 17.3 years, 555 males/ 824 females) were recruited between January and June 2014. The study population included 95 individuals previously known as hepatitis C positive. Serums samples were collected and screened for the detection of IgG anti-HAV, HBsAg and HBcAb by Elisa Test. The overall anti- HAV seroprevalence was about 84.7%. There was no statistically significant difference between male and female. On the 1379 tested individual, 219 were positive for HBcAb and 67 were positive for HBsAg. IgG anti- HAV were positive in 80.6% of HBsAg-positive patients (54 out of 67), 81.3% of HBcAb-positive patients (178 out of 219) and in 95.8% of HCV-positive patients (91 out of 95). HBV infection and HCV infection were statistically associated with a greater risk of positive anti-HAV antibody (p< 0.001). Our study revealed that Thala represents an intermediate endemicity level and that the introduction of vaccination against HAV in this region is recommended, especially for the hepatitis B or C infected person seronegative for HAV.