Thymol action on cells and tissues of the synganglia and salivary glands of Rhipicephalus sanguineus sensu lato females (Acari: Ixodidae).

Thymol is a monoterpene present in plants of the families Lamiaceae, Verbenaceae and Apiaceae. Despite its proven acaricidal activity, little is known about the mechanism of action of thymol in ticks. Thus, the aim of this study was to perform a morpho-histochemical analysis of the synganglion and salivary glands of partially engorged females of the brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.), exposed to thymol at different concentrations. Five groups were established: Control Group I (distilled water), Control Group II (ethanol 30%), Group III (thymol 1.25 mg/mL), Group IV (thymol 2.5 mg/mL) and Group V (thymol 5.0 mg/mL). The females were exposed to the treatments by the immersion method and subsequently kept in a climatic chamber (27 ± 1 °C and relative humidity 80 ± 10%) for five days. After this period, the synganglion and salivary glands were removed, and the hematoxylin/eosin morphological technique was applied. The von Kossa staining method with counterstaining neutral red was performed on the salivary glands. The results showed that females exposed to thymol had damaged synganglia, with pyknotic nuclei and vacuoles in the cortex and subperineurial regions, as well as rupture of the neural lamellae. The salivary glands showed type I acini with a dilated lumen. Cells with extremely vacuolated cytoplasm and fragmented nuclei were observed in type II and III acini. Type II acini of the females exposed to thymol revealed different calcium staining when compared to the Control Groups I and II. We therefore conclude that the salivary glands and synganglion are subject to changes in morphology and calcium levels when exposed to thymol at concentrations of 1.25, 2.5 and 5.0 mg/mL, demonstrating that this monoterpene has acaricidal potential on partially engorged females of R. sanguineus (s.l.).

Evaluation of the acaricidal activity of thymol incorporated in two formulations for topical use against immature stages of Rhipicephalus sanguineus sensu lato (Latreille, 1806) (Acari: Ixodidae).

The objective of this study was to assess, for the first time, the in vitro acaricidal activity of two topical formulations containing thymol, on immature stages of Rhipicephalus sanguineus sensu lato. For this purpose, two base formulations were prepared: an oil-in-water (O/W) emulsion and a hydroalcoholic solution, containing different thymol concentrations (0.5 to 20 mg/mL). We used the larval packet test for non-engorged larvae and nymphs, and the immersion test for engorged larvae and nymphs. For emulsion, a mortality rate of 94.2% was achieved at 0.75 mg/mL in non-engorged larvae. For engorged larvae, there was 95.0% mortality at 5.0 mg/mL. Non-engorged nymphs showed 83.3% mortality at 2.5 mg/mL, and for engorged nymphs, 86.0% mortality was verified at 5.0 mg/mL. For the hydroalcoholic solution, the mortality found for non-engorged larvae was 88.1% at 2.5 mg/mL. For engorged larvae, the highest mortality was 25.0% at 20 mg/mL; non-engorged nymphs had 91.0% mortality at 1.0 mg/mL and for engorged nymphs; the maximum value verified was 18.3% mortality at 20 mg/mL. Preliminary stability tests were carried, and the hydroalcoholic solution remained stable under all the conditions analyzed. The O/W emulsion showed signs of early instability at the concentration of 5.0 mg/mL. The results obtained indicate that the acaricidal activity of thymol, when included in the proposed formulations, was enhanced against non-engorged larvae with topical treatment in comparison with data in the literature. Although there were variations in toxicity between the different stages, these formulations are promising for future therapeutic use.