The bioactive compound carvacrol as a potential acaricide: An assessment of its effects on the integument of female Rhipicephalus sanguineus sensu lato ticks.

Studies seeking control methods for infestation of Riphicephalus sanguineus sensu lato (s.l.) ticks (dog ticks) have been carried out in order to minimize damage to both the tick's hosts and the environment, the latter due to the misuse of acaricide products. In this regard, carvacrol has been used as a natural alternative against ticks as it displays several properties including acaricidal. In this context, the present study aimed to evaluate the ultramorphology and morphohistochemistry of the integument of semi-engorged R. sanguineus s.l. females exposed to different carvacrol concentrations. The findings indicate that the integument surface of females exposed to the highest carvacrol concentration (25 µl/ml) became wrinkled, suggesting dehydration or the result of integument cuticular and epithelial layer disorganization in response to the toxic product. Morphohistochemical integument layer alterations were more significant and intense in females exposed to the highest carvacrol concentration (25 µl/ml), confirming dose-dependent carvacrol action. Among other cell and tissue alterations, changes in epithelial cell shape, size, and arrangement (epidermal layer) were noted, alongside altered and pyknotic-shaped nuclei, suggesting a death process for these cells. This epithelium changed from simple cubic to stratified, also in response to the presence of the evaluated bioactive compound. Thus, the findings reported herein demonstrate that carvacrol may be an alternative for an efficient and more sustainable tick control in the near future.

Sublethal concentrations of acetylcarvacrol affect reproduction and integument morphology in the brown dog tick Rhipicephalus sanguineus sensu lato (Acari: Ixodidae).

Rhipicephalus sanguineus sensu lato (s.l.), also known as the brown dog tick, is among the main tick species involved in the transmission of pathogens to humans and other animals and, therefore, the target of numerous control methods. However, due to the disadvantages of synthetic acaricides, the use of alternative products such as plant derivatives has been encouraged. This study aimed to evaluate the acaricidal potential of acetylcarvacrol and to determine its efficacy at sublethal doses for the control of R. sanguineus s.l. female ticks. In addition, as acetylcarvacrol was applied topically, morphological alterations in the integument were assessed. Acetylation of carvacrol was performed by reaction with acetic anhydride in a sodium hydroxide solution, being confirmed by infrared spectroscopy. The lethal concentration for 50 and 90% (LC50 and LC90) of unfed ticks and the efficacy of acetylcarvacrol in engorged females were determined after the Adult Immersion Test (AIT). For the evaluation of effects of acetylcarvacrol in the integument, routine histological techniques were employed after the AIT. The LC50 and LC90 in unfed females were 2.8 and 7.2 µL/mL, respectively. Regarding reproductive performance, after treatment with 8.0 µL/mL acetylcarvacrol 90.9% control was achieved, as ticks showed the lowest egg production index (EPI), hatching rate (HR), and fecundity rate (FR). In the integument, considerable morphological alterations were observed both in cuticle and epithelium. Thus, acetylcarvacrol affected R. sanguineus s.l. external coating and reproduction when applied at sublethal concentrations, probably contributing to a long-term control.

Low concentrations of acetylcarvacrol induce drastic morphological damages in ovaries of surviving Rhipicephalus sanguineus sensu lato ticks (Acari: Ixodidae).

Rhipicephalus sanguineus sensu lato (s. l.) ticks are targets of acaricidal treatments due to their great veterinary and medical importance. Acetylcarvacrol stands out as a promising acaricidal substance for its increased biocidal activity and stability. Additionally, its toxicity to mammals is reduced when compared to the parent molecule, carvacrol. The present study aimed to evaluate the effects of acetylcarvacrol on the morphology of ovaries of engorged R. sanguineus s. l. ticks. The animals were subjected to the Adult Immersion Test to calculate the lethal concentrations (LC50 and LC90) of acetylcarvacrol. Subsequently, the surviving ticks were dissected for collection of the ovaries. The samples were processed through routine histological techniques. The histological sections were stained with hematoxylin and eosin, bromophenol blue (protein detection) and PAS (polysaccharide detection). The alterations found in the ovaries were evaluated by morphometric and semiquantitative analysis. The LC50 and LC90 were 17.805 and 26.164 µL/mL, respectively. The most severe morphological alterations were disappearance of the female germ cells (oocytes), nucleolus vacuolization, thicker and irregular chorion, and decrease in size of the oocytes. Also, the content of proteins and carbohydrates in the oocytes were heavily affected by the chemical, as evidenced by a non-homogeneous staining pattern. The group exposed to the highest concentration of acetylcarvacrol (20 µL/mL) exhibited a statistically greater score in the semiquantitative analysis when compared to the other groups. The morphological changes in the ovaries may reduce the ticks' offspring production or generate descendants that will struggle to carry out essential biochemical processes during their lives. Thus, acetylcarvacrol may be a promising alternative to control tick infestation by impairing the reproduction of this parasite.

Effects of carvacrol on oocyte development in semi-engorged Rhipicephalus sanguineus sensu lato females ticks (Acari: Ixodidae).

Currently, the most commonly used method to control ticks is the use of synthetic acaricides. However, these compounds are potentially harmful to hosts and the environment, in addition to causing the selection of resistant individuals. Therefore, several studies have been conducted to find sustainable methods to control ticks, such as Rhipicephalus sanguineus sensu lato, the most important vectors of pathogens for dogs. There has been increasing research on natural products with acaricidal action, especially with regard to plant-derived compounds as carvacrol, an aromatic monoterpene with several biological properties, including repellent and acaricidal activities, besides exerting cytotoxic effects on the exposed ticks. The objective of the present study was to evaluate the action of different carvacrol concentrations on the ovaries morphophysiology in semi-engorged R. sanguineus s.l. females to determine its effect on oocyte development. The results showed the occurrence of significant morphological alterations in the shape of oocytes (from round-shaped to irregular) and in the germinal vesicles, in addition to extensive cytoplasmic vacuolation.These effects were observed after the application of carvacrol at a concentration of 20 µL/mL. The most significant alterations were observed at the highest concentration (100 µL/mL), at which the oocytes could not develop further than stage II (total absence of oocytes III, IV and V). These data showed that even though carvacrol was unable to kill all ticks at these concentrations, surviving females could have had an altered reproduction, which would hinder the generation of new individuals, resulting in a long-term control. Data regarding the inhibition of oocyte development are unprecedented and indicate the use of carvacrol as a natural product with the potential to control R. sanguineus s.l. ticks.

The effects of neem oil (Azadirachta indica A. JUSS) enriched with different concentrations of azadirachtin on the integument of semi-engorged Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) females.

Several studies searching for methods to control Rhipicephalus sanguineus s.l., (dog tick) infestations have been developed aiming to minimize the damages caused by these ectoparasites to the hosts and the environment, which is harmed by the indiscriminate use of toxic acaricide products. In this scenario, neem oil has been used as a natural alternative against ticks, once this chemical has repellent properties and interferes in the growth regulation of these ectoparasites, inhibiting ecdysis. The present study evaluated the effects of azadirachtin-enriched neem oil on the integument of semi-engorged R.sanguineus s.l., females through morphohistological techniques. The results showed the occurrence of significant morphological and histochemical alterations, mainly in the females exposed to higher concentrations, which demonstrates the dose-dependent action of the chemical. A decrease in the cuticle thickness was observed, as well as a modification in the distribution of the epithelial cells, which displayed pyknotic and fragmented nuclei, and intensely vacuolated cytoplasm, indicating that these cells would be undergoing death processes. These morphological alterations observed in the integument of the females exposed to the azadirachtin-enriched neem oil encourage the use of this chemical as a strategy to control these ectoparasites.