In this study, we investigated the response of freshwater oligochaete, Tubifex tubifex, to the water temperature changes and the differences of substrate composition in a laboratory condition. The changes of body shape were observed in a test cage according to the water temperature change ranging from 10℃ to 30℃ with 2℃ interval every 10 minutes. The substrate preference was observed with four different substrate composition from silt-clay to coarse sand. Our results displayed that T. tubifex preferred substrates with the smallest particle size (<0.063 mm). The water temperature influenced on the activity and body shape of T. tubifex, showing low activity with the coiled and constricted body shapes at lower temperature and high activity with relaxed linear body shapes at higher temperature.