Amebiasis is cosmopolitan in its distribution both in developed and developing countries. The aim of this study was to assess the prevalence of Entamoeba hitolytica (E. histolytica) since 2007 to May 2017 at Mulu Assefa Hospital in Atsbi Wonberta Woreda, Eastern Tigray Ethiopia. This study showed that the prevalence rate of E. histolytica within ten years ranged from 15.08% to 72.60%. Out of these stools examined 6195(46.64%) were infected with E. histolytica with annual prevalence rate of 46.64%. Of these the prevalence rate of E. histolytica was 2913 (21.93%) for males and 3282 (24.71%) for females. The result showed that there was highest prevalence in spring 1983 (14.92%), followed by summer 1981 (14.91%), winter 1291 (9.71%) and least in autumn season 1184 (8.91%). The overall prevalence rate of the disease within the ten years, among different age groups in young (21-40 years) was highest 2175 (16.38%), followed by adult (≥ 41 years) 1863 (14.03%), moderate in child (0-11 years) 1250 (9.41%) and least in adolescence (12-19 years) 1043 (7.85%) out of 13283 total stool examined. The result of prospective laboratory assessment of five month (January to May 2017) on patient attended in the Hospital, showed that totally 685 patients were stools examined in the Hospital, out of these 226 were E. histolytica positive. There was higher prevalence in females than males. There was highest prevalence in young and least in adult and the overall prevalence was 32.99%. The prevalence of E. histolytica had association between sex, season and age groups at p<0.05. As the respondents answered 84% of them were have knowledge on the mode of transmission, prevention method and risk factors that contribute for transmission of the disease. However, 16% the people had less awareness toward the disease. Therefore, protection of water sources from contamination and health education are needed to improve sanitation, personal hygiene and a proper usage of toilet to reduce infections of E. histolytica.