The liquid from rumen’s cattle is expected to be used as nutrients in microalgae culture. This study aimed to examine the use of waste of rumen’s cattle as a source of nutrients with different doses in the culture of Thalasiossira sp. The research design used was a completely randomized design (CRD) with 5 treatments with 3 replications of (A) 1 ml conwey/l of seawater, (B) 1 ml of waste/l of sea water, (C) 10 ml of waste/l of seawater, (D) 1 ml of waste + 0.4 gr NaN03/l of seawater, and (E) 10 ml of waste + 0.4 gr NaN03/l of seawater. The calculated parameter were the population density of Thalasiossira sp., diatom diameter, protein content and water quality. The results showed that the waste of rumen’s cattle could be used as an alternative source of nutrients in the culture of Thalasiossira sp. and the peak of the population was reached on day 3. Measurement of protein content ranged from 9.9 to 11.8% and the diameter of the diatoms ranged from 5-10 m. The measurement of the water quality of the media was in the range of temperature 24-26 oC, pH value 7 and salinity 32-34 ppt.