The probiotic powder can improve the value of hematology, reduce blood fat and increase the production of late-phase laying hens. The study was conducted to assess the impact of probiotic powder on the different physiological activities system of late-phase laying hens measured with hematological values and serum biochemistry. The research was carried out from March until April, 2022 at the Layer Farm Sukarapi Village, Universitas Padjadjaran, Indonesia. A total of 40 laying hens 90 weeks were randomly allocated to four treatments and five replications, with experimental Completely Randomized Design (CRD). The treatments were, T0: probiotic powder free-control diet. T1: ration and 2 % powder probiotic, T2: ration and 3% powder probiotic, T3: ration and 4% powder probiotic. Statistically, there was no significant differences (P>0.05) in hemoglobin, hematocrit, erythrocyte, leukocyte, cholesterol, and triglyceride blood levels. Meanwhile, our data showed statistically significant difference in glucose blood levels. However, the administration of powder probiotics of 3% and 4 % has improved the hemoglobin, hematocrit, erythrocyte, and leukocyte levels, and decreased cholesterol, and triglyceride blood levels.