The excessive use of acaricides in the management of the tick Rhipicephalus (Boophilus) microplus has favored the development of resistance in the parasites, in addition to causing environmental contamination. In this context, biological control using entomopathogenic nematodes appears as a promising alternative. This study evaluated the efficacy of two isolates, Heterorhabditis bacteriophora (HB EN01) and Steinernema rarum (PAM 25), in the in vitro control of R. (B.) microplus. Two experiments were conducted with different concentrations of nematodes. In the first, fifteen engorged females were exposed to suspensions containing 100, 200 and 300 infective juveniles (IJs) per female. The second experiment was carried out in soil substrate, with a concentration of 200 IJs per female. Three application systems were tested: S1 (females placed on the substrate before application), S2 (suspension applied before the introduction of females) and S3 (females placed in a voile bag on the substrate). For S. rarum (PAM 25), there was no significant difference in oviposition inhibition, but larval hatching was lower at concentrations of 200 and 300 IJs/female compared to 100 (p < 0.05). H. bacteriophora (HB EN01) showed a significant reduction in oviposition only between concentrations of 100 and 300 IJs/female (p < 0.05), with no variation in larval hatching or efficacy. On the soil substrate, both isolates showed efficacy above 80 %, except for S. rarum in the voile system (60 %). Treated females showed darkening and dehydration from the 4th day onwards. The control group remained healthy. The results indicate that EPNs are an effective alternative in the control of R. (B.) microplus.
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