Seketak River is a river in the upstream region at Tembalang. The area which was previously an open area for water absorption, has changed into the campus and residential areas. As a result, community needs for facilities, especially water, increased. Therefore, blocking the Seketak river to built Diponegoro reservoirs was planned as an effort to conserve water and to control the destruction by water force. Macrobenthos group is a group of animals that are relatively settled in the bottom waters and often used as biological indicator of water quality. The research were implemented on June-July 2012. The purpose of this study were determine the waters condition of the Station I, II, and III seketak river based on macrobenthos and find out the abundance, diversity, and uniformity of macrobenthos at Seketak River. The research were determined by survey method to collect information from representated population. The results of this research on the station I obtained individuals abundance value was 589 ind./m2, diversity index was 1,14, and uniformity index is 0,45; Station II individual values obtained individuals abundance value was 273 ind./m2, diversity index was 0,83, and uniformity index was 0,40; Station III obtained individuals abundance value was 319 ind./m2, diversity index was 1,01, and uniformity index was 0,57. The results of this research indicated that the water condition of the Station I and III was in the medium to heavily polluted category and water condition of the Station II was in the heavily polluted category