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In vitro and in vivo activities of a biocontrol agent, Bacillus stearothermophilus strain YC4194 was evaluated for the control of Pythium damping-off of cucumber. B. stearothermophilus YC4194 inhibited germination of cystospores and formation of zoosporangia of Pythium aphanidermatum in vitro. Incorporation of a bentonite and talc based formulation(109 cfu/g) of B. stearothermophilus YC4194 to the nursery soils (10 g/l soil) resulted in a significant (p=0.01) reduction in the disease severity of cucumber damping-off after inoculation with P. aphanidermatum. The control efficacy of B. stearothermophilus YC4194 formulation was not different from that of the fungicides, dimethomorph, metalaxyl, ethaboxam. When the cucumber plants were transplanted to the soil inoculated with P. aphanidermatum zoospores, the B. stearothermophilus YC4194 maintained the high population density in rhizosphere soil upto 107 cfu/g until 15 days after treatment.