The acaricidal effect of 14 strains of Metarhizium anisopliae sensu lato isolated from soil of livestock farms in the Mexican tropics was evaluated against larvae and engorged females, and during the laying and hatching of eggs of Rhipicephalus microplus (Ixodida: Ixodidae). For each fungal strain, the larvae mortality percentage was evaluated through a larval immersion test, while the reproductive efficiency indices in engorged females were measured using adult immersion tests at a dose of 1 × 10⁸ conidia/ml. All strains of M. anisopliae (s.l.) proved to be highly effective against R. microplus larvae (66–100%) and engorged females (100%). The strains also showed a good effect in inhibiting egg laying (16.45–56.38%) and a moderate effect in decreasing egg hatching (5.24–32.68%). Two strains demonstrated to be effective against all development phases of R. microplus in an integrated manner.