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To establish the optimal ratios of chicken manure extract (CME) and suspending yeast solution (SYS) for rearing rotifers, the population density, population growth rate, fatty acid compositions of rotifers and the unionized ammonia (NH 3 -N) and total phosphorus (TP) concentrations in cultured water under different ratios of CME and SYS (BG, EG1, EG2, EG3, EG4 and EG5) were investigated. The results showed that the population density and population growth rate of rotifers in EG2 (4 ml•L −1 CME: 0.3 ml•L −1 SYS) were the highest and reached 387.33 ± 12.34 and 0.57 ± 0.01 ind.•ml −1 , respectively. In addition, Pearson's correlation analysis indicated that the proportions of polyunsaturated fatty acids (PUFAs), n-3PUFAs, n-6PUFAs and long-chain PUFAs in rotifers were positively correlated with the content of CME (p < 0.05). Furthermore, NH 3 -N and TP in the cultured water decreased with the SYS contents increasing.Compared with EG5, the concentrations of NH 3 -N and TP in EG2 were reduced by 58.83 ± 0.05% and 72.52 ± 0.17%, respectively. These results indicated that both the population growth and fatty acid composition of rotifers, and the concentrations of NH 3 -N and TP in the cultured water were significantly affected by the ratios of CME and SYS. And EG2 was optimal for rearing the rotifer.