This study was carried out to provide preliminary data for increasing biodiversity in agricultural ecosystem and investigate availability of using Tubificidae spp. for environment-friendly agriculture through the survey of growth of Tubificidae spp. in flooded paddy fields during winter. We estimated the number of Tubificidae spp. in rice paddy where had been flooded during the previous winter in Ganghwa-gun, Suwon-si, and Hwaseong-si (Bongdam-eup and Paltan-myeon) between March and April from 2010 to 2012. During growing period of rice, the number of Tubificidae spp. was compared between flooded and non-flooded paddy field in Ganghwa-gun. As results, mean density of Tubificidae spp. in winter flooded paddy field was 7,235.5 $ind./m^2$ in Ganghwa-gun, 14,347.5 $ind./m^2$ in Suwon-si, 59,989.9 $ind./m^2$ in Bongdam-eup, and 2,610.6 $ind./m^2$ in Paltan-myeon. Mean density of Tubificidae spp. was the highest in flooded paddy fields of Bongdam-eup that was kept wet until rice harvest time and had shallow water. While mean density was the lowest in Paltan-myeon where had deep water and later flooded paddy field than others. In Ganghwa-gun, during growing period of rice, mean density of Tubificidae spp. in flooded paddy fields (171,109.0 $ind./m^2$) and flooded fallow paddy fields (321,084.2 $ind./m^2$) was much higher than non-flooded paddy fields (1,006.6 $ind./m^2$). Flooded paddy field during winter can help increase the number of Tubificidae spp. which can control weeds and enhance biodiversity in paddy fields for environment-friendly agriculture. Early flooding and shallow water of paddy fields could be a good agriculture practice to encourage the growth of Tubificidae spp..