BACKGROUND: Biofortification through Pseudomonas putida IFO 14796 intervention has succeeded to improve iron content in maize grain up to 18.79%. AIM: This study was carried out to assess the effect intake of maize biofortified iron (MBI) on the red blood cell (RBC) formation rate in Wistar rat anemia induced 2,4 dinitrophenylhydrazine. METHODS: Randomized complete design was carried out to assess the effect of MBI on the erythrocyte level with four levels of MBI treatment (R1=10%; R2=12%; R3=14%, and R4=16% of rat body weight [BW]) and one of control (Ro=10% maize non-biofortified iron), and five replications, respectively. Erythrocyte level was measured using the Counting Neubauer Chamber Method after Wistar anemia induced 2,4-dinitrophenylhydrazine (DPNH) after intake MBI for 7 days. Data were analyzed by ANOVA and Fisher’s least significant difference. RESULTS: There was an influence intake of MBI level on the improving rate the erythrocyte formation in Wistar anemia (p < 0.05). Treatment R1 improved to 0.058±0.034% significantly different with R0 (0.006±0.017%), but non-significantly with R2 and R4 at p < 0.05. Linear regression showed the equation, y=−0.002x+0.07; R2=0.69. CONCLUSION: Intake MBI more than 10% BW negatively effect to RBC formation rate of Wistar rat’s anemia.