Effects of Piper aduncum (Piperales: Piperaceae) Essential Oil and Its Main Component Dillapiole on Detoxifying Enzymes and Acetylcholinesterase Activity of Amblyomma sculptum (Acari: Ixodidae).

Amblyomma sculptum is a species of tick in the family Ixodidae, with equids and capybaras among its preferred hosts. In this study, the acaricidal activity of the essential oil (EO) from Piper aduncum and its main component, Dillapiole, were evaluated against larvae of A. sculptum to establish lethal concentration values and assess the effects of these compounds on tick enzymes. Dillapiole exhibited slightly greater activity (LC50 = 3.38 mg/mL; 95% CI = 3.24 to 3.54) than P. aduncum EO (LC50 = 3.49 mg/mL; 95% CI = 3.36 to 3.62) against ticks. The activities of α-esterase (α-EST), ß-esterase (ß-EST), and glutathione-S-transferase (GST) enzymes in A. sculptum larvae treated with Dillapiole showed a significant increase compared to the control at all concentrations (LC5, LC25, LC50 and LC75), similar results were obtained with P. aduncum EO, except for α-EST, which did not differ from the control at the highest concentration (LC75). The results of the acetylcholinesterase (AChE) activity show an increase in enzyme activity at the two lower concentrations (LC5 and LC25) and a reduction in activity at the two higher, lethal concentrations (LC50 and LC75) compared to the control. These results suggest potential mechanisms of action for these natural acaricides and can provide guidance for the future development of potential plant-derived formulations.

Enzootic stability of tick fever in Holstein calves grazing in a tropical region, subjected to strategic cattle tick control with fluralaner.

BACKGROUND: In 2022, fluralaner was launched on the market for use in the control of the cattle tick Rhipicephalus microplus after showing 100% efficacy in registration trials against the causative agents of cattle tick fever (TFAs). The aim of the present study was to determine whether a strategic control regimen against R. microplus using fluralaner (FLU) in Holstein calves grazing in a tropical region would alter the enzootic stability status of cattle tick fever, triggering outbreaks in these animals up to 22 months age. METHODS: In this study, a group of calves treated with FLU was compared with a control group treated with the regimen currently being used on the farm, which consisted of the fipronil + fluazuron formulation (FIFLUA). In the first experiment, the efficacy of the FIFLUA pour-on formulation was evaluated in a field study. In the second experiment, which lasted 550 days, two experimental groups (n = 30/group) of Holstein calves naturally infested with R. microplus were analyzed. Calves aged 4 to 10 months received either a specific treatment regimen with FLU (experimental group) or FIFLUA (control group). During this period, tick counts, animal weight measurement, feces collection (to determine eggs and oocysts per gram of feces), tick fever monitoring, blood smears (to ascertain enzootic stability of the herd), PCR testing for TFAs and serology (indirect enzyme-linked immunosorbent assay [iELISA]) were performed. All calves were evaluated for signs of tick fever between ages 11 and 22 months. RESULTS: FIFLUA showed an acaricidal efficacy of > 90% from post-treatment days 14 to 35. Regarding treatments against the TFAs, the average number of treatments was similar between groups, but animals treated with FLU had a smaller reduction in packed cell volume on some of the evaluation dates of the second and third treatment against TFAs. In calves aged 10 months in the FLU group, B. bovis was not detected by PCR (0/15 samples), 40% of the samples had antibody titers and 33% (10/30) of the samples had positive blood smears. Regarding B. bigemina, > 86% of the samples in both groups tested positive for B. bigemina DNA and antibodies; there was no difference in the antibody titers between the groups. There were no clinical cases of cattle tick fever in calves aged 11 to 22 months. CONCLUSIONS: In comparison with the control treatment, the strategic control regimen against R. microplus with FLU that was implemented in the present study did not negatively affect the enzootic stability status of A. marginale and B. bigemina in the herd up to 22 months of age. The enzootic stability status of B. bovis was not reached by either group. These results likely represent a characteristic of the local tick population, so further studies should be performed.

Alterations in the metabolism of Pseudosuccinea columella (Mollusca: Gastropoda) caused by Heterorhabditis bacteriophora HP88 (Rhabditida: Heterorhabditidae).

The gastropod Pseudosuccinea columella participates in the dissemination of Fasciola hepatica in the environment, acting as the main intermediate host of this parasite in Brazil. The present study sought to elucidate the possible pathogenic effects of the entomopathogenic nematode (EPN) Heterorhabditis bacteriophora on P. columella, by evaluating the influence of infection on alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as well as the concentrations of total protein, uric acid, and urea in the snail's hemolymph. For this, the snails were exposed to EPNs for 24 and 48 h, and for each exposure time, 20 snails were dissected after 7, 14 and 21 days for hemolymph collection. The primary findings suggest a significant proteolysis alongside elevated levels of uric acid and urea in P. columella infected individuals. These findings reveal that H. bacteriophora HP88 infection induced serious changes in the snail's metabolism, triggering important deleterious effects.

Efficiency of Experimental Formulation Containing Duddingtonia flagrans and Pochonia chlamydosporia against Moniezia expansa Eggs.

This study aimed to evaluate the effectiveness of the experimental formulation containing chlamydospores of Duddingtonia flagrans and Pochonia chlamydosporia fungi, against Moniezia expansa. Two experiments were carried out. The first experiment evaluated the in vitro efficacy using 1 g of the experimental formulation (V1) added to 100 M. expansa eggs and the control (V2) (without the fungal formulation). Intact eggs or eggs with alterations were counted in order to evaluate their effectiveness. The second experiment evaluated the action of the fungal formulation on M. expansa eggs after passing through the gastrointestinal tract of goats. Three groups were identified as B1, B2, and B3, which received 1.0, 1.5 g of experimental fungal formulation, and placebo, respectively. In experiment 1, all the eggs in V1 were subjected to the predatory action of fungi, while in V2, the eggs remained intact. In experiment 2, the reduction of eggs in groups B1 and B2 were 49% and 57% 24 h after ingestion, 60% and 63% 48 h after, and 48% and 58% 72 h after. The predatory capacity against M. expansa eggs shown in the tests demonstrated that experimental fungal formulation has the potential to be used on integrated helminth control programs.

Do Commercial Insect Repellents Provide Protection against the Tick Amblyomma sculptum (Acari: Ixodidae)?

Amblyomma sculptum is a species of public health interest because it is associated with the transmission of the bacteria that causes Brazilian Spotted Fever (BSF). The use of repellents on humans is a prophylactic measure widely used to provide protection against a series of arthropod vectors, including mosquitoes and ticks. However, in Brazil, the effectiveness of commercial repellents against A. sculptum is little known. Therefore, it is necessary to carry out specific studies to evaluate the repellency of these commercial products, registered for use against mosquitoes, against the star tick. The main goal of the present work was to evaluate the effectiveness of six commercial repellents against A. sculptum. Unfed nymphs, aged between two and eight weeks, were tested against products based on DEET (OFF!® and Repelex®), Icaridin (Exposis® and SBP®), and IR3535 (Johnsons and Henlau). Three bioassays were conducted to evaluate tick behavior: (i) filter paper, (ii) fingertip, and (iii) field. All bases tested showed high repellent activity, differing significantly (p < 0.05) from the control. It was observed the formulation with DEET resulted in the best results in the filter paper bioassay. In the fingertip bioassay, the DEET base repelled a greater number of ticks compared to Icaridin. In the field bioassay, there was no significant difference between the Icaridin base and DEET, and both formulations differed from the control (p < 0.05). The six formulations tested showed significant percentages of repellency against ticks; however, for the fingertip and field bioassays, the products OFF!®, Repelex®, and Exposis® were tested as they showed better performance in the filter paper test. OFF!® showed the best percentage of repellency (100%), followed by Repelex® (96.8%), and Exposis® (93.1%), considering the two-hour period of the bioassay-field-test. Proving the effectiveness of repellents on the market against A. sculptum presented in this study is crucial, since this is the main ectoparasite of humans that can transmit Rickettsia rickettsii when infected. The effectiveness of commercial insect repellents against other tick species that parasitize humans can also be explored.

Number of rainy days in a week influencing screwworm navel myiasis in beef calves and efficacies of injectable and topical antiparasitics.

Rainfall incidence as a risk factor for umbilical myiasis by Cochliomyia hominivorax in newborn beef cattle and the preventive and curative efficacies of injectable and topical products against its larvae were evaluated. The prevalence of navel myiasis did not differ between sex, however, it did differ between seasons of the year, independent of animal sex, with it being higher in the rainy season than in the dry season for both sexes (males 64.6%, females 62.1%). During the rainy season, rains occurred intermittently and the number of rainy days in a week had a direct influence on prevalence, with an increase (≥87%) in the number of calves with navel myiasis caused by C. hominivorax. When it rained intermittently for four, five and six days in a week during the rainy season, calves had ≈12, 24 and 11 times greater chances, respectively, of having navel myiasis compared to a week without rain during the rainy season. The injectable products did not reach preventive efficacies higher than 35%, while all treatments of topic products reached 100% preventive efficacy at 10 days post-treatment. Only the formulations with 30% dichlorfenthion and 0.32% fipronil achieved 100% curative efficacy. The results reinforce the need for intensified inspection and healing of the umbilicus during the rainy season, mainly during rainier weeks. Furthermore, topical formulations may be a better alternative as a preventive treatment than avermectin formulations. However, the curative efficacy of the topical product containing dichlorvos did not demonstrate a relationship with preventive efficacy for newborn calves.

Acaricidal activity of carvacrol and thymol on acaricide-resistant Rhipicephalus microplus (Acari: Ixodidae) populations and combination with cypermethrin: Is there cross-resistance and synergism?

This study evaluated the acaricidal activity of thymol and carvacrol on Rhipicephalus microplus populations with different resistance profiles and investigated the synergistic effect of combinations of these monoterpenes with cypermethrin. The adult immersion test (AIT) was used to characterize the susceptibility of tick populations (45 field populations) to synthetic acaricides: deltamethrin, amitraz and chlorfenvinphos. The larval packet test (LPT) was used to determine the LC50 values for thymol (25 tick populations) and carvacrol (20 tick populations). The susceptible strain Porto Alegre (POA) was used as a reference for calculating the resistance ratio (RR). Subsequently, larval immersion tests (LIT) were performed with combinations of cypermethrin with thymol or carvacrol to assess a synergistic effect. In the AIT, deltamethrin showed efficacy > 90% in one (2.2%) population tested (mean: 12.1 and 11.1 for populations 1-25 and 26-40, respectively), whereas amitraz and chlorfenvinphos showed efficacy > 90% for two (4.4%) populations (mean: 61.3 and 47.3 for populations 1-25 and 26-40, respectively) and eight (17.7%) populations (mean: 69.7 and 59.7 for populations 1-25 and 26-40, respectively). In the LPT, the LC50 values for thymol and carvacrol varied from 0.67 to 2.12 mg/mL and 0.55-3.21 mg/mL, with an average LC50 for populations of 1.49 and 1.75 mg/mL, respectively. For thymol, no resistance was observed in any of the populations, values of RR50 > 1.5. There was no correlation between the LC50 values for thymol and the efficacy of the chemical acaricides tested. Regarding carvacrol, for only one tick population had the value of RR50 > 1.5, indicating an incipient resistance. No correlation was observed between the LC50 values for carvacrol and the efficacy of tested acaricides. The combination of thymol and carvacrol with cypermethrin showed a synergistic effect in the resistant population (Jaguar - thymol 4.19 and carvacrol 3.67), and no synergistic interaction were showed in the susceptible population. Answering the questions we conclude that: 1 - The comparison between the LC50 values for thymol and carvacrol in field populations and the susceptible strain POA suggests the absence of cross-resistance (ticks and terpenes), and the differences between the LC50 values for thymol and carvacrol in the different R. microplus populations are inherent to the characteristics of each population tested; 2 - the combination of thymol or carvacrol with cypermethrin showed a synergistic effect with different activity according to the population of ticks.

Susceptibility of embryos of Biomphalaria tenagophila (Mollusca: Gastropoda) to infection by Pochonia chlamydosporia (Ascomycota: Sordariomycetes).

Schistosoma mansoni is a heteroxenous parasite, meaning that during its life cycle needs the participation of obligatory intermediate and definitive hosts. The larval development occurs in aquatic molluscs belonging to the Biomphalaria genus, leading to the formation of cercariae, which emerge to infect the final vertebrate host. For this reason, studies for control of the diseases caused by digenetic trematodes often focus on combating the snail hosts. Thus, the objective of this study was to evaluate the susceptibility of Biomphalaria tenagophila embryos to the fungus Pochonia chlamydosporia (isolate Pc-10). The entire experiment was conducted in duplicate, with five replicates for each repetition (five egg masses/replicate), utilizing a total of 100 egg masses, with 20-30 eggs/egg mass. At the end of 15 days, the egg masses were evaluated under a stereomicroscope to analyze the hatching of B. tenagophila embryos in both experimental groups. After days of interaction, the exposure to the fungal hyphae bodies significantly impaired the viability of the B. tenagophila eggs, inhibiting the embryogenesis process by 83.7% in relation to the control group. Transmission and scanning electron microscopic images revealed relevant structural alterations in the egg masses exposed to the hyphae action of the fungus, interfering in the development and hatching of the young snails under analysis. These results indicate the susceptibility of B. tenagophila embryos to the fungus P. chlamydosporia (isolate Pc-10) and suggest the potential of Pc-10 to be used in the control of intermediate host, for its ovicidal capacity and for being an ecologically viable option, but in vivo experiments become necessary.

Techniques for monitoring dairy calves against the tick fever agents: a comparative analysis.

Data regarding parasitemia (blood smears), rectal temperature (RT), packed cell volume (PCV) and vaginal mucosa coloration (VMC) of Gyr x Holstein female calves between 3-7mo were accessed to evaluate different techniques for monitoring the bovine tick fever agents (TFA). The 1st experiment determined the correlation between the TFA parasitemia with RT and PCV. The 2nd, evaluated the associated risk of A. marginale parasitemia with RT and PCV in relation to the Gyr/Holstein genetic proportion (5/8,3/4,7/8 and 15/16) using Receiver Operating Characteristic Curve (ROC). The 3rd, two groups were performed: cattle monitored by RT (T01) and by PCV (T02), during their 80-210 days of age, data regarding TFA parasitemia, RT, PCV, VMC and weight were registered. In 1st experiment, RT showed weak correlation with TFA parasitemia, while PCV showed a strong correlation with A. marginale and B. bigemina, but not with B. bovis parasitemia. In experiment 2, the ROC curve analysis showed that when the genetic proportion of B. t. taurus increased, least reliable RT was to monitor calves infected with A. marginale. The PCV for monitoring A. marginale was the best technique, showing sensitivity of 74.2% and specificity of 97.0% than other techniques that used RT and VCM as a monitoring tool. In general, calves monitored by PCV (T02) showed higher PCV values, lower A. marginale parasitemia, less pneumonia as co-infection and less salvation treatment were performed than in animals monitored by RT (T01). Furthermore, animals from T02 gained 23.5 kg more than those from T01. The low frequency of B. bovis and B. bigemina found in this study made impossible to compare the monitoring techniques for these pathogenic agents.

Effect of the formulation with fipronil and fluazuron on the reproductive biology and ovaries histopathology of Rhipicephalus microplus engorged females.

This study aimed to evaluate the effects of a commercial formulation containing fipronil and fluazuron on the reproductive biology and the morphology of ovaries from Rhipicephalus microplus engorged females. To carry out the study, three calves were artificially infested every 3 days with approximately 5000 larvae. On day 0, the animals were treated with a commercial formulation containing fipronil (1.25 mg/kg) + fluazuron (2.5 mg/kg). Before the application of the acaricide, engorged females of R. microplus were collected to constitute the control group (10 for biology analyses and 20 for histology analyses). After applying the commercial formulation, naturally detached engorged females were recovered on days + 5, + 10, and + 20 (10 engorged females/day) to evaluate their reproductive biology, and on days + 4, + 12, and + 20 (20 engorged females/day) for histological evaluation of the ovaries. Females from the treated groups produced smaller amounts of eggs, exhibiting lower viability when compared to eggs from the control group (p < 0.05). The ovaries of females from all treated groups (+ 4, + 12, and + 20) showed morphological changes, including: cytoplasmic disorganization, cytoplasmic degradation, irregular shape of the oocyte and germinal vesicle, reduction and vacuolization of yolk granules and oocyte disruption. Oocytes were observed in smaller numbers in all stages of development (I, II, III, IV, and V) and greater numbers of indeterminate oocytes were verified in the ovaries of the treated groups when compared to the control group. Therefore, results showed that the commercial formulation containing fipronil and fluazuron affected the reproductive biology, caused morphological changes in the ovaries, and reduced the number of oocytes in R. microplus engorged females.

Spatial distribution and risk factors for sheep toxoplasmosis in Goiás, Brazilian Cerrado Region.

The present study aimed to evaluate the spatial distribution and risk factors for infection by Toxoplasma gondii in sheep in the state of Goiás, located in the central-western region of Brazil. Through the immunofluorescence antibody test (IFAT), the seroprevalence of anti-T. gondii antibodies was analyzed in 1000 blood serum samples obtained from sheep in all macro and micro regions of the state of Goiás. Data related to sex, age of the animals, size of the farm, type of farm, water source, veterinary assistance, replacement of the herd, presence of domestic cats, presence of wild cats and presence of other wild animals were obtained at the sampling time. The differences between the seroprevalences obtained in relation to the variables analyzed were estimated using Pearson's chi-square test (χ2). The odds ratio (OR) values for each risk factor evaluated were statistically analyzed with a confidence interval of 95%. Positivity for IgG anti-T. gondii was observed (titer ≥64) in 34.3% (343/1000) of the samples, which ranged from 26.9% (31/115) to 44.2% (53/120) and from 21.8 (12/55) to 55.2% (16 / 29), respectively in the analyzed mesoregions and microregions. In all investigated regions of the State of Goiás, serum-reactive animals were detected with the age of the animals, the source of water, the form of replacement of the herd and the presence of domestic cats and wild animals risk factors statistically associated with the occurrence of T. gondii in animals.

Evaluation of rotational grazing as a control strategy for Rhipicephalus microplus in a tropical region.

Rhipicephalus microplus is the most significant tick of livestock and its control is particularly challenging due to its resistance to commercial acaricides. Pasture rotation is considered a management strategy that could help control R. microplus, however, the literature only contemplates mathematical models and little is known about the effects of this practice in the field. The objective of this work was to determine whether pasture rotation is an efficient method for controlling R. microplus. Two different experiments were performed that involved groups of continuous and rotational grazing bovines. Female ticks measuring 4.5-8.0 mm were counted on animals while larvae in pasture were counted using the flannel drag technique. Treatment for infested bovines was applied when the average group tick count was ≥30 females. The results showed that rotational grazing (with 20-day periods of rest) had a higher tick count on-host than continuous grazing (P < 0.05) and additional bovine treatment was needed. Sixty and 105 days were needed to re-infest and disinfest pasture of R. microplus larvae, respectively. The first treatment of bovines occurred 91 days after the animals were placed in a closed area. The results indicate that rotational grazing is not an efficient way to control R. microplus.

Entomopathogenic nematodes in pharmaceutical formulations for Rhipicephalus microplus (Acari: Ixodidae) control: In vitro evaluation of compatibility, thermotolerance, and efficiency.

This work aimed to investigate candidate biological control agents against the cattle tick Rhipicephalus microplus by studying three entomopathogenic nematodes (EPN) isolates from two species, Heterorhabditis bacteriophora (isolates LPP30 and HP88) and Heterorhabditis baujardi (isolate LPP7). The study comprised the pharmacotechnical development of four test formulations, and their biological evaluation, in three stages: 1) selection of the fittest EPN isolate at 16 ± 1 °C after 120 h of incorporation in the formulations; 2) determination of the effects of temperature and incubation time on the survival of the selected isolate in formulations; 3) evaluation of the in vitro efficacy (infectivity) of the selected EPN formulations and their components against R. microplus females after 72 h exposure. H. baujardi LPP7 presented the highest survival rates in 0.1% carboxymethyl cellulose sodium (CMC) formulations (100% at 25 °C, up to 10 days after incorporation). At the highest tested temperature (34.2 °C), 54% of LPP7 survived for up to 72 h in CMC formulations. The infectivity of LPP7-containing formulation against R. microplus was 99%, reducing oviposition, egg production index and larval hatching. Considering the pressing need for safer acaricide products, incorporation of EPNs in pharmaceutical formulations is presented as a promising approach for the biological control of cattle tick infestations, potentially reducing environmental impact and resistance development associated with chemical acaricides.

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.).

In vitro assessment of the acaricidal activity of carvacrol, thymol, eugenol and their acetylated derivatives on Rhipicephalus microplus (Acari: Ixodidae).

This study reports the comparative evaluation of the activity of carvacrol, thymol, eugenol and their respective acetylated derivatives (carvacrol acetate, thymol acetate and eugenol acetate) on Rhipicephalus microplus, to verify the possible influence of the acetate group. The acetylated derivatives were prepared from reactions of the phenolic compounds with acetic anhydride/pyridine. The formation of the products was confirmed by analysis of hydrogen and carbon-13 nuclear magnetic resonance (1H and 13C NMR) spectra. The larval packet test was used to evaluate the acaricidal activity, with concentrations of 0.312, 0.625, 1.25, 2.5, 5.0, 7.5, 10.0 and 15.0 mg/mL. Thymol and carvacrol resulted in 100% mortality starting at the concentration of 2.5 mg/mL, while the same was observed for carvacrol acetate starting at the concentration of 5.0 mg/mL. For the other treatments, 100% mortality was only achieved in the groups treated with the highest concentration (15.0 mg/mL). The LC50 and LC90 values (mg/mL) of carvacrol acetate (2.49, 4.21), thymol acetate (2.97, 8.52) and eugenol acetate (4.25, 13.10) were higher than those for the corresponding precursor molecules carvacrol (0.83, 2.02), thymol (1.26, 2.21) and eugenol (2.77, 5.35). The acetylation process did not enhance the activity of these substances on unengorged larvae of R. microplus, since the precursor substances (carvacrol, thymol and eugenol) had greater efficacy.

Chemical composition and acaricidal activity of the essential oil of Baccharis dracunculifolia De Candole (1836) and its constituents nerolidol and limonene on larvae and engorged females of Rhipicephalus microplus (Acari: Ixodidae).

Baccharis dracunculifolia DC (common name "alecrim-do-campo" in Brazil) is a plant with widespread distribution in South America that is the botanical origin of green propolis. The aim of this study was to evaluate the chemical composition and acaricidal activity of the essential oil of B. dracunculifolia and its constituents nerolidol and limonene on unengorged larvae and engorged females of Rhipicephalus microplus (Acari: Ixodidae). The essential oil yield was 0.8% of dry mass and the major constituents were nerolidol (22.3%), germacrene D (7.2%), limonene (6.9%), ß-pinene (6.7) and bicyclogermacrene (6.5%). The acaricidal activity of the essential oil and the pure compounds nerolidol and (R)-(+)-limonene were assessed in the laboratory through the modified larval packet test (LPT) and the female immersion test (FIT). In the LPT, the essential oil and nerolidol were both active, causing more than 90% mortality at concentrations from 15.0 and 10.0 mg mL(-1), respectively, whereas (R)-(+)-limonene was not active. In the FIT, the oil and nerolidol caused reduction in the quantity and quality of eggs produced, with control percentages of 96.3% and 90.3% at concentrations of 60.0 and 50.0 mg mL(-1), respectively. It can be concluded that the essential oil obtained from the aerial parts of B. dracunculifolia and its major component nerolidol have high activity on R. microplus larvae and engorged females.

Assessment of the acaricidal activity of carvacrol, (E)-cinnamaldehyde, trans-anethole, and linalool on larvae of Rhipicephalus microplus and Dermacentor nitens (Acari: Ixodidae).

The acaricidal activity of carvacrol, (E)-cinnamaldehyde, trans-anethole, and linalool was studied on Rhipicephalus microplus and Dermacentor nitens larvae. All the substances were tested at concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 µl/ml, with 10 repetitions per treatment. The modified larval packet technique was employed in the tests and the mortality was evaluated after 24 h. In the groups treated with carvacrol, the lowest concentration (2.5 µl/ml) was sufficient to cause 100% death of the R. microplus and D. nitens larvae. The same concentration of (E)-cinnamaldehyde resulted in death of approximately 99% of the larvae of both tick species and reached 100% at the other concentrations. For trans-anethole, mortality rates above 90% of the R. microplus and D. nitens larvae were only observed starting at the concentration of 15.0 µl/ml and reached 100% at the highest concentration (20.0 µl/ml). Finally, the mortality rates of the groups treated with linalool were low, only reaching 8.4 and 14.5% at the highest concentration (20.0 µl/ml) for larvae of D. nitens and R. microplus, respectively. These results show that carvacrol, (E)-cinnamaldehyde, and trans-anethole have acaricidal activity, particularly carvacrol and (E)-cinnamaldehyde, both of which resulted in high mortality rates for the larvae of these two tick species even at the lowest concentration.

Activity of essential oil of Lippia triplinervis Gardner (Verbenaceae) on Rhipicephalus microplus (Acari: Ixodidae).

The objective of this work was to characterize and investigate the acaricidal activity of the essential oil of the aerial parts of Lippia triplinervis at different concentrations on unengorged larvae and engorged females of Rhipicephalus microplus. The essential oil yielded 2.21 % (w/w to dry matter) and was composed mainly of carvacrol (31.9 %), thymol (30.6 %), and p-cymene (12.3 %). Two tests were performed to assess the acaricidal activity: the modified larval packet test, with concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 mg/mL and the female immersion test, with concentrations of 10.0, 20.0, 30.0, 40.0, and 50.0 mg/mL. There were ten repetitions for each concentration, and for each test, a control group was formed in which the ticks were treated with Tween 80 (20 mg/mL). The experimental groups were kept in a climate-controlled chamber (27 ± 1 °C and RH >80 %). The mortality of the larvae was above 95 % in all the groups tested and reached 100 % as of the 5.0 mg/mL concentration, while the control group exhibited 0 % mortality. In the female immersion test, there was a significant decline (p < 0.05) in the egg mass weight, egg production index, and hatching percentage starting at the concentration of 30.0, 40.0, and 20.0 mg/mL, respectively, and the control percentage at the concentrations of 40.0 and 50.0 mg/mL were above 90 and 95 %. The L. triplinervis oil as thus an alternative source of the monoterpenes thymol, carvacrol, and p-cymene, and its toxicity on R. microplus larvae and females makes it promising possibility for control of this tick.

Enhancing the acaricide effect of thymol on larvae of the cattle tick Rhipicephalus microplus (Acari: Ixodidae) by solubilization in ethanol.

The aim of the present work was to propose a new method of solubilizing thymol for use as an acaricide and to evaluate the resulting product's efficacy on Rhipicephalus microplus larvae. Two experiments were performed, both employing the larval packet test to evaluate the potential of the solutions formulated. In the first experiment, R. microplus larvae were exposed to three new formulations: formulation 1-distilled water (70%) + ethanol (30%) + thymol (10.0 mg/ml); formulation 2-distilled water (50%) + ethanol (50%) + thymol (10.0 mg/ml); and formulation 3-distilled water (55%) + ethanol (40%) + DMSO (5%) + thymol (10.0 mg/ml). A control was prepared for each formulation (same formulations except thymol), and another control group was formed where the larvae did not receive any treatment. From the results obtained in the first experiment, the formulation with the best performance and cost-benefit relation was chosen and was tested at concentrations of 2.5, 5.0 and 10.0 mg/ml on larvae of this tick. In the first experiment, the mortality of the groups treated with thymol was greater than 95% for all three formulations, with significant differences (p < 0.05) in comparison with the control groups, where the mortality was under 1%. In the second experiment, formulation 2 was selected because of its better cost-benefit relation, and the mortality rates were 94.0%, 96.5% and 99.9% for the concentrations of 2.5, 5.0 and 10.0 mg/ml, respectively, demonstrating that thymol dissolved in ethanol has high acaricide activity on cattle tick larvae.

Acaricidal activity of the essential oils from Eucalyptus citriodora and Cymbopogon nardus on larvae of Amblyomma cajennense (Acari: Ixodidae) and Anocentor nitens (Acari: Ixodidae).

The present study evaluated the acaricidal activity of essential oils from Eucalyptus citriodora and Cymbopogon nardus on non-engorged larvae of Amblyomma cajennense and Anocentor nitens. In order to carry out the study, six groups were formed, each concentration being a treatment (6.25%, 12.5%, 25%, and 50%, respectively) and also with the creation of a control group (distilled water) and a positive control (Deltametrine). For each treatment, approximately 100 larvae of these ticks were placed onto filter papers (2 x 2 cm) impregnated with the concentrations used to test. Next, the envelopes were closed bearing inside the filter paper with measurements of 6 x 6 cm. For each group, six repetitions were performed, and after 24 h live and dead larvae were counted. This procedure was carried out for two essential oils on the two species of ticks. For A. cajennense, the acaricide efficacy of E. citriodora oil was of 10.8%, 35.3%, 34.5%, and 53.1%, whereas the efficacy of C. nardus was of 0.0%, 0.0%, 0.0%, and 61.1% at concentrations of 6.25%, 12.5%, 25.0%, and 50.0%, respectively. In relation to A. nitens, the acaricide efficacy of E. citriodora oil was of 20.1%, 84.5%, 89.2%, and 100.0%, whereas the efficacy of C. nardus was of 0.0%, 90.8%, 100.0%, and 100.0% at concentrations of 6.25%, 12.5%, 25.0%, and 50.0%, respectively. The results indicate that the essential oils tested showed a promising acaricidal activity mainly on A. nitens larvae.