: Sterilization in dental practice is crucial for infection prevention. The aim of this study was to identify the presence and causes of bacterial growth using biological indicators in dental sterilization equipment in San Luis Potosí, S.L.P., Mexico, with different consecutive measurements over a year. : This longitudinal cohort, conducted from January 2022 to January 2024 in San Luis Potosí, Mexico, aimed to identify the presence and causes of bacterial growth in dental sterilization equipment using biological indicators. A total of 207 dental offices were approached, and 175 participated, providing data through questionnaires and monitoring sterilization cycles with BIs. The checks were bimonthly for one year, with a total of six checks. : (a) An 11% ( = 1188) incidence of bacterial growth was observed, with a higher percentage in dry heat equipment (13%). (b) Upon analyzing the six consecutive verifications over a year, no statistically significant differences were observed in the failures of the sterilization cycles when comparing the two pieces of equipment. (c) Error in temperature and time of the equipment (OR = 4.0, 95% CI = 1.6-3.9, = 0.0001) was significantly associated with the presence of bacterial growth during the one-year period. : Monitoring sterilization cycles and identifying the causes of bacterial growth with different consecutive verifications decreased the presence of bacterial growth (failures) during the one-year period.