摘要:The validation of sterilization processes in food and pharmaceutical industries is a major tool for quality assurance, making the products safe, effective and reliable. Biological indicators, formed by spores of Bacillus stearothermophilus microorganisms considered at higher thermal resistance, are used to evaluate and monitor the physical parameters of a cycle of steam sterilization. In this way this study aimed to cultivate and characterize the microorganism Bacillus stearothermophilus generations, assessing the influence of these generations in the parameters of resistance, formation and concentration of its spores to be used in the production of biological indicators. The experiments were conducted cultivating the 1st, 2nd and 3rd generations of B. stearothermophilus in suitable culture media for sporulation, in Roux bottles, for a period of 15 days of incubation. During these 15 days, the sporulation process was evaluated by microscopy, according to Wirtz-Conklin's method, every 24 hours of incubation. The results showed that the generations do influence the sporulation process, indicating that the 3rd generation is the most suitable for the production of formed spores, in concentration and thermal resistance appropriate characteristics to the needs of a biological indicator to efficiently validate and monitor steam sterilization cycles. http://dx.doi.org/10.14685/rebrapa.v1i1.12