Combination of cypermethrin and thymol for control of Rhipicephalus microplus: Efficacy evaluation and description of an action mechanism.

The cattle tick Rhipicephalus microplus is one of the most important ectoparasites in the tropical and subtropical regions of the world. Synthetic pyrethroids are widely used to control this tick, and the selection of resistant populations is a huge problem worldwide. The activity of thymol, a natural monoterpene, free or in combination with other compounds, has been demonstrated against different species of ticks. However, the mode of action is not fully understood. The present study aimed to evaluate the efficacy and the potential mode of action of the combination of cypermethrin and thymol on ticks from two populations with different levels of susceptibility to cypermethrin (low and high susceptibility). The isolated acaricidal activity of cypermethrin and thymol on larvae was carried out in different concentrations. The combination with different concentrations of cypermethrin and fixed concentrations of thymol (1300 µg/mL for the low susceptibility population; 690 µg/mL for the high susceptibility population) were performed. Adult engorged females were divided into five experimental groups (n = 20): 1) Control group untreated; 2) Control group: 2.0% (v/v) DMSO; 3) Thymol group: 1300 µg/mL thymol; 4) Cypermethrin group: 3700 µg/mL cypermethrin; 5) Association of cypermethrin (3700 µg/mL) + thymol (1300 µg/mL). A subgroup was used to study the efficacy of the reproductive parameters and another subgroup, with ten adults from each treatment, was used to quantify thymol and cypermethrin by HPLC chromatographic analysis. All compounds tested were effective on larvae from both populations, and the combination with thymol decreased the LC of cypermethrin (232.4 to 52.7 µg/mL) on the low-susceptibility population. The combination of thymol and cypermethrin was effective in both populations of R. microplus (reproductive performance of engorged females) when compared to the untreated control group, even with higher percent control values (pop. 1: 93.5 ± 5.6% and pop. 2: 92.7 ± 1.1%) than the group treated only with cypermethrin (pop. 1: 87.3 ± 7.3% and pop. 2: 83.5 ± 1.2%). From the HPLC analyzes, a higher concentration of cypermethrin (pop. 1: 30.3 ± 6.9 and pop. 2: 45.4 ± 17.7 ng/mg) was detected in the tissues of engorged females treated with the combination compared to analyte concentrations in groups treated with cypermethrin only (pop. 1: 12.4 ± 4.4 pop. 2: 25.5 ± 9.4 ng/mg). This was the first study to investigate the acaricidal efficacy of the combination of thymol + cypermethrin on R. microplus and demonstrate that the presence of thymol increases the concentration of cypermethrin in the internal tissues of engorged females through a possible mechanism for increasing the penetration of cypermethrin at the cuticular level.