Terpenes on Rhipicephalus (Boophilus) microplus: Acaricidal activity and acetylcholinesterase inhibition.

The Rhipicephalus (Boophilus) microplus tick is the main ectoparasite of cattle in tropical and subtropical regions worldwide. Resistance to chemical acaricides has become widespread affirming the need for new drugs to tick control. Terpenes have become a promising alternative for cattle tick control, however the mechanism of action of these compounds is still controversial. Inhibition of acetylcholinesterase (AChE) is a well established mechanism of action of organophosphate and carbamate acaricides, but the possible action of terpenes on tick AChEs has seldom been studied in resistant and sensitive strains of R. (B.) microplus. The aim of the present study was to evaluate terpene inhibition of AChE from resistant and sensitive strains of R. (B.) microplus in correlation with their acaricidal activity. Among the terpenes used in the present study, p-cymene, thymol, carvacrol, and citral displayed acaricidal activity with LC50 of 1.75, 1.54, 1.41, and 0.38 mg.mL-1 for the susceptible strain, and LC50 of 1.40, 1.81, 1.10, and 1.13 mg.mL-1 for the resistant strain. Thymol and carvacrol inhibited the AChE of the susceptible strain larvae with IC50 of 0.93 and 0.04 mg.mL-1, respectively. The IC50 exhibited by eucalyptol, carvacrol and thymol for AChE of the resistant strain larvae were 0.36, 0.28, and 0.13 mg.mL-1, respectively. This was the first study to investigate the action of terpenes on AChE from susceptible and resistant R. (B.) microplus. As not all terpenes with acaridical activity showed AChE inhibition, the participation of AChE in the acaricidal activity of terpenes needs further investigation.

The first assessment of the stress inducible defense of Leucaena leucocephala with acaricidal potential effect against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

Plants respond to wounding caused by mechanical stress or herbivory by synthesizing defense proteins. There are no studies reporting the action of induced plant proteins against ticks. The aim of this study was to investigate the effect of mechanically wounded Leucaena leucocephala leaves against Rhipicephalus (Boophilus) microplus. Initially, we carried out time course experiments to evaluate the impact of mechanical wounding on the protein content and the peroxidase, catalase and protease inhibitor activities in L. leucocephala. We then evaluated the acaricidal activity on R. (B.) microplus from protein extract collected from L. leucocephala after mechanical wounding. L. leucocephala leaves were artificially wounded, and after 6, 12, 24 and 48h, the leaves were collected for protein extraction. Quantitative and qualitative analyses of the proteins were performed. The protein content and peroxidase and protease activities increased 12h after wounding, and the acaricidal activity of this protein extract was evaluated using engorged R. (B.) microplus females. The protein extract obtained after wounding reduced egg production (8.5%) compared to those without wounding. Furthermore, the extract reduced egg hatching by 47.7% and showed an overall efficacy of 56.3% at 0.1 mgP/mL of the protein. We demonstrated that L. leucocephala defensive proteins could be effective against R. (B.) microplus.

Acaricidal activity of Lippia gracilis essential oil and its major constituents on the tick Rhipicephalus (Boophilus) microplus.

The present study aimed to evaluate the activity of Lippia gracilis Schauer essential oil obtained from different L. gracilis genotypes and their major components, carvacrol and thymol against Rhipicephalus (Boophilus) microplus (cattle tick) larvae and engorged females. The larval test was performed parallel to the adult immersion test for engorged females for four L. gracilis genotypes. Similar tests were further performed for their major compounds carvacrol and thymol. Carvacrol (LC50 of 0.22 and 4.46 mg/mL, to larvae and engorged females, respectively) was more efficient than thymol (LC50 of 3.86 and 5.50 mg/mL, to larvae and engorged females, respectively). The lethal concentrations obtained for the isolated essential oil from genotypes LGRA-201 against larvae (1.31 mg/mL) and LGRA-106 against engorged females (4.66 mg/mL) confirmed the acaricidal activity of L. gracilis essential oil and its effectiveness in controlling the southern cattle tick.

Sero-prevalence and risk indicators for canine ehrlichiosis in three rural areas of Brazil.

Ehrlichia canis has a worldwide geographic distribution, occurring particularly in tropical and subtropical areas. In Brazil, the main vector in urban areas is believed to be the brown dog tick Rhipicephalus sanguineus, but little is known about the occurrence, transmission and other epidemiological aspects of canine ehrlichiosis in rural areas, where Amblyomma ticks are found more frequently than R. sanguineus. A sero-prevalence study of canine ehrlichiosis was carried out in three distinct rural regions of the State of Minas Gerais, Brazil. Serum samples were collected from 226 dogs living on farms in Lavras (n=85), Belo Horizonte (n=45), and Nanuque (n=96) and were analyzed by an indirect fluorescent antibody test for the detection of anti-Ehrlichia canis antibodies. Age, breed, sex, presence of ticks and packed cell volume were also recorded. There were 65.6% positive dogs in Nanuque, 37.8% in Belo Horizonte, and 24.7% in Lavras. Animals living in Nanuque were 4.6 times more likely to be serologically positive than dogs living in the other two regions and antibody titres were considerable higher in this area. Male dogs, dogs >5 years of age, those infested with ticks, and mongrels all showed higher rates of positivity. The results point to the importance of canine ehrlichiosis in rural areas and indicate the need for further studies on natural transmission and maintenance of the disease.