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Musca domestica L., 1758, is an important mechanical vector of several pathogens for humans and livestock, making it essential to study new alternatives of more efficient and safer control for this dipteran. This study evaluated the toxicity of the phenylpropanoids eugenol and (E)-cinnamaldehyde on its life stages. A contact test with 10 repetitions (n = 10) was performed for each substance concentration on each post-embryonic immature life stage. Both substances presented insecticidal activity on the immature life stages of the dipteran, and secondary effects on development caused by sublethal concentrations. Larvicidal activity was shown from the 1.25 mg/ml concentration by eugenol and from 2.5 mg/ml by (E)-cinnamaldehyde, and both substances had a 100% larval treatment efficacy (LTE) from the 5mg/ml concentration. For pupal treatment, (E)-cinnamaldehyde differed from the control from the 10 mg/ml concentration (P < 0.05), and both phenylpropanoids caused malformation in adults from 10 mg/ml. The highest pupal treatment efficacy (PTE) was obtained from the 30 mg/ml concentration, 67.2% for (E)-cinnamaldehyde, and 32% for eugenol. The products tested in this study showed high larvicidal potential, and both presented pupicidal effects and caused malformation in adults from treated pupae., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Thymol is a monoterpene with proven acaricidal activity on different tick species and life stages. The objective of this work was to evaluate the effect of thymol on the reproductive biology of engorged females of Rhipicephalus sanguineus sensu lato and the morphological changes caused in the cells of the tubular and accessory glands of Gené's organ. The females were exposed to thymol by immersion. Seven groups were established: group I (distilled water), group II (30% ethanol), group III (thymol 1.25 mg/mL), group IV (thymol 2.5 mg/mL), group V (thymol 5.0 mg/mL), group VI (thymol 10.0 mg/mL), and group VII (thymol 20.0 mg/mL), with 20 replicates for each treatment. The ticks were kept in a BOD incubator at 27 ± 1 °C and RH > 80 ± 10%. Ten females from each group were evaluated daily until death, and the remaining ten were kept in the incubator under the same conditions for five days and then dissected for the removal of Gené's organ to note possible damage to cell morphology by scanning electron microscopy (SEM) and hematoxylin-eosin (HE) techniques. Thymol demonstrated 63% and 98% efficacy in groups VI (10.0 mg/mL) and VII (20.0 mg/mL), respectively. In these groups, the cells of the tubular and accessory glands of Gené's organ showed signs of damage: irregular eosin staining, rupture and deformation of the cellular limit, presence of fragmented nuclei, changes in cytoplasmic homogeneity and areas with deformation (folds) in the basal region., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

The acaricidal activity of combinations of thymol, carvacrol and eugenol was evaluated on larvae and engorged females of the cattle tick Rhipicephalus microplus. The first step assessed the compounds separately, in concentrations of 3.125, 6.25, 12.5 and 25 mg/mL. Then tests were performed with the compounds combined in the ratio of 1:1 at concentrations of 3.125 and 6.25 mg/mL, along with the control group treated with the solvent (3% DMSO). In the second step, combinations were tested incorporated in a formulation at the concentration de 6.25 mg/mL, using the larval packet and adult immersion tests. The associations carvacrol + thymol (3.125 mg/mL), carvacrol + eugenol and thymol + eugenol (6.25 mg/mL) presented synergism, while the other associations had an additive effect. In the experiments with formulation, all combinations caused 100% larval mortality, but the efficacy was under 15% against engorged females. Therefore, the combinations of thymol + carvacrol (3.125 mg/mL) as well as carvacrol + eugenol and eugenol + thymol (6.25 mg/mL) had a synergistic effect on engorged females, but when incorporated in the formulation, the acaricide activity was strong against larvae but weak against engorged females., (Published by Elsevier Inc.)

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.)., (Copyright © 2018. Published by Elsevier GmbH.)

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., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

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 ( 1 H and 13 C 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 LC 50 and LC 90 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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study evaluated the acaricidal activity of the methanol extract of Acmella oleracea with 0.187% of spilanthol against immature stages of Amblyomma sculptum. The packet test was used to evaluate the extract's activity on unengorged larvae and nymphs, testing concentrations of 0.4 to 50 mg/mL for larvae and 12.5 to 200.0 mg/mL for nymphs. For the engorged stages, the immersion test was used, at concentrations of 0.4 to 50 mg/mL for larvae and 12.5 to 200.0 mg/mL for nymphs. The methanol extract caused 100% mortality of the unengorged larvae and nymphs starting at concentrations of 12.5 and 200.0 mg/mL, respectively. For engorged larvae and nymphs, the mortality was 100% starting from concentrations of 12.5 and 150.0 mg/mL, respectively. The LC 50 for unengorged larvae was 3.2 mg/mL, while for engorged larvae it was 6.6 mg/mL. For unengorged nymphs, the LC 50 was 38.5 mg/mL, but it was not possible to calculate the corresponding value for engorged nymphs because the data did not fit the probit model. These results demonstrate that the methanol extract of A. oleracea has acaricidal activity against different immature stages of A. sculptum., (Copyright © 2018. Published by Elsevier B.V.)

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.

The current study evaluated parasitism of wild birds by ticks in a fragment of Atlantic Forest in the state of Minas Gerais, Brazil, and examined the ticks for rickettsial agents. Birds were captured during 2014 and 2015 and ticks were identified by sequencing fragments of the 16S and 12S ribosomal DNA. Among 260 birds representing 19 families and 52 species, a total of 69 (26.5%) were found to be infested by larvae (LL) and/or nymphs (NN) of Amblyomma longirostre (Koch, 1844) (45 LL, 4 NN), Amblyomma calcaratum Neumann, 1899 (9 LL, 15 NN), Amblyomma nodosum Neumann, 1899 (2 NN), Amblyomma parkeri Fonseca and Aragão, 1952 (21 LL), Amblyomma sp. haplotype Nazaré (77 LL), and Haemaphysalis leporispalustris (Packard, 1869) (17 LL, 1 NN). The use of PCR and sequencing of the rickettsial genes gltA, htrA, ompA and ompB, revealed the presence of "Candidatus Rickettsia amblyommii" in A. longirostre (13/49; 26%) and Rickettsia parkeri (strain ApPR) in both A. parkeri (1/21; 5%) and haplotype Nazaré (42/77; 55%) ticks. In addition, we detected Rickettsia rhipicephali in 31 (40%) of the 77 haplotype Nazaré ticks. This is the first record of this rickettsial agent in a species of the genus Amblyomma. The pathogenic potential of this bacterium is undetermined, but the unprecedented association with Amblyomma ticks may represent a cause for concern for public and/or animal health., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

We evaluated the acaricidal activity of Acmella oleracea methanol extract and spilanthol on Rhipicephalus microplus and Dermacentor nitens. The extract was made through maceration with methanol. From this extract, a dichloromethane fraction with 99% spilanthol was obtained and tested on R. microplus larvae and engorged females and D. nitens larvae. For evaluation against larvae, the modified larval packet test was used, and both the methanol extract and dichloromethane fraction were tested at concentrations of 0.2-50mg/mL. The modified larval packet test was also used in the lethal time (LT) test, with the methanol extract at a concentration of 12.5mg/mL and the percentage mortality was assessed after 15, 30, 45, 60, 75, 90, 120min and 24h. The 50% lethal time calculation (LT 50 ) was performed in this test. The engorged female test was performed with R. microplus only, at concentrations of 25-200mg/mL for methanol extract and 2.5-20.0mg/mL for spilanthol. The methanol extract caused 100% mortality of the R. microplus and D. nitens larvae at concentrations of 3.1 and 12.5mg/mL, respectively. Spilanthol resulted in 100% mortality of R. microplus larvae at concentration of 1.6mg/mL and of D. nitens at 12.5mg/mL. In the lethal time assay using the methanol extract, the mortality rate was 100% for R. microplus and D. nitens larvae after 120min and 24h, with LT 50 values of 38 and 57min, respectively. In the test of females, the egg mass weight and the hatching percentage of the groups treated with concentrations equal to or higher than 50.0mg/mL of methanol extract were significantly reduced (p<0.05), while for spilanthol, the reduction of the egg mass weight and hatching percentage occurred from concentrations of 10.0mg/mL and 2.5mg/mL, respectively. Females treated with 200.0mg/mL of extract died before starting oviposition, resulting in 100% effectiveness, while the best efficacy for spilanthol was 92.9% at a concentration of 20.0mg/mL. Thus we conclude that the methanol extract of A. oleracea and spilanthol have acaricidal activity against R. microplus and D. nitens., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Birds are an important component of the life histories and bioecology of a number of tick species and of some tick associated pathogens. An examination of the data concerning bird/tick associations in the Neotropics, showed that the tick Haemaphysalis leporispalustrisis (Packard, 1869) was rarely recorded infesting birds. The current study reports parasitism by H. leporispalustris in wild birds collected from Atlantic rain forest environments in the states of Rio de Janeiro (4LL) and Minas Gerais (17LL, 1NN), Brazil. All ticks were identified morphologically to the genus level; total DNA was extracted from each Haemaphysalis tick and examined by PCR and nucleotide sequencing of fragments of the eukaryotic genes encoding 16S rRNA and 12S rRNA. The bird species Arremon semitorquatus, Corythopis delalandi, Fluvicola nengeta, Troglodytes musculus, and Volatinia jacarina were recorded as hosts for H. leporispalustris for the first time in South America, and Turdus rufiventris represented a new record for Brazil.

Haemoproteid species have a wide global distribution, and they have been described in falcon species in several parts of the world. However, few studies in South America have focused on these birds. Haemoproteus spp. infections have been reported as the causative agents of serious histopathological changes, which can lead to the death of the host. Thus, this study aimed to molecularly and phylogenetically characterize Haemoproteus spp. in Caracara plancus, to characterize aspects of parasitism through clinical analysis and biochemical parameters, and to describe the histopathology of infection. To examine these aspects, 5 southern caracaras were examined clinically, and blood samples were collected. Blood smears were subsequently utilized in parasitemia calculations, PCR amplification, and serum biochemical investigations. Histological sections of the liver, kidneys, spleen, and heart were examined to check for possible histopathological changes. The birds showed clinical signs such as pallor and prostration that are consistent with Haemoproteus spp. infection. Moreover, the examination of the blood smears revealed 0.07% parasitemia in young gametocytes only. The PCR and sequencing results confirmed that the parasites belonged to Haemoproteus spp. The activity of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) enzymes, albumin, total serum proteins, and enzymatic urea were first described in C. plancus and serve as reference for future studies of bird species parasitized by Haemoproteus spp. Histopathology results showed signs of injury that were consistent with haemosporidian infection in the tissues of the analyzed organs. The present study is preliminary, and additional studies of Haemoproteus spp. infections in other bird species are needed to better understand the relationship between parasites and hosts, because despite the low parasitemia recorded, biochemical and histopathological changes in various organs were observed.

The Rickettsia bacteria include the aetiological agents for the human spotted fever (SF) disease. In the present study, a SF group Rickettsia amblyommii related bacterium was detected in a field collected Amblyomma sculptum (Amblyomma cajennense species complex) tick from a Brazilian SF endemic site in southeastern Brazil, in the municipality of Juiz de Fora, state of Minas Gerais. Genetic analysis based on genes ompA,ompB and htrA showed that the detected strain, named R. amblyommii str. JF, is related to the species R. amblyommii.

The habits of birds make them more or less susceptible to parasitism by certain tick species. Therefore, while some bird species are typically found to be intensely infested, others are relatively unaffected. This study investigated the occurrence of ticks in Passeriformes inhabiting an Atlantic Forest fragment in southeastern Brazil, during the dry and rainy seasons, by means of parasitological indexes and multiple correspondence analysis, to determine the factors that influence tick parasitism in these birds. Data were collected on 2391 ticks, all classified in the Amblyomma genus, from 589 birds. The ticks identified to the species level were A. longirostre, A. nodosum, A. calcaratum, A. parkeri, and A. ovale. Thamnophilidae, Conopophagidae, Thraupidae, Dendrocolaptidae, and Platyrinchidae were the families with the highest prevalence. In terms of parasite intensity, the families Conopophagidae, Thamnophilidae, Thraupidae, Furnariidae, and Pipridae stood out with the highest values. Bird species that are generalists regarding eating habits and habitat occupation tended to have higher parasite loads, as did larger species and those inhabiting the understory. The tick prevalence was higher in the dry season than in the rainy season. The majority of the ticks were collected from the head region, mainly around the eyes and in the nape. Also, this work reports 22 new bird-parasite relations.

This study aimed at assessing the combined effect of thymol, carvacrol and (E)-cinnamaldehyde on Amblyomma sculptum and Dermacentor nitens larvae. The effects resulting from treatments were evaluated by means of the modified larval packet test. In order to determine the LC50, components of essential oils, the monoterpenes thymol, carvacrol and phenylpropanoid (E)-cinnamaldehyde were individually tested at different concentrations. After determining the LC50, each essential oil component was separately evaluated and then combined with another substance at a 1:1 proportion at the LC50 concentration and at 1/2 and 1/4 of the LC50. For A. sculptum, the lowest LC50 value was obtained for (E)-cinnamaldehyde (1.40 mg/ml), followed by thymol (2.04 mg/ml) and carvacrol (3.49 mg/ml). The same order of effectiveness was observed for D. nitens, with values of 1.68, 2.17 and 3.33 mg/ml, respectively. In the evaluation of component associations of essential oils against A. sculptum larvae, only the combinations between carvacrol and thymol (LC50) and carvacrol and (E)-cinnamaldehyde (1/4 LC50) presented a moderate synergetic effect. In turn, for D. nitens larvae, the combinations between thymol and carvacrol (LC50 and 1/2 LC50) presented a synergetic effect, while the others presented an additive or antagonistic effect. Therefore, it can be concluded that the combination of thymol and carvacrol (LC50) has a moderate synergetic effect against A. sculptum larvae, while thymol, combined with carvacrol (LC50 and 1/2 LC50), has a synergetic effect against D. nitens larvae., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This is the first study to investigate the activity of thymol on Rhipicephalus microplus larvae under semi-natural conditions. For this purpose, tests were conducted in pots with Brachiaria decumbens seedlings containing cattle tick larvae. Thymol, diluted in ethanol 50° GL, was tested at concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 mg/mL, along with the control group treated with the solvent alone. Each treatment was composed of five pots (1 pot = a repetition). The experiment was performed in three steps. On the first day, the larvae were applied at the base of the signalgrass. Twenty-four hours later, approximately 25 mL of the solution was applied with thymol on the top of the vegetation in each pot. The survival of the larvae was measured 24 h after application of the solutions. Each pot was analyzed individually, and the grass fillets contained larvae were cut with scissors, placed in Petri dishes, and taken to the laboratory to count the number of living larvae. At the highest concentrations (10, 15, and 20 mg /mL), the number of live larvae declined by more than 95 % in relation to the control group. The lethal concentration 50 % (LC50) and LC90 values were 3.45 and 9.25 mg/ml, respectively. The application of thymol in semi-natural conditions starting concentration of 10 mg/mL significantly reduced the number of living R. microplus larvae.

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%), β-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., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The cattle tick, Rhipicephalus (Boophilus) microplus, has caused serious harm to livestock raising in Brazil, considering the costs of controlling it, loss of revenue due to smaller production of milk and meat, and damage to leather, in addition to transmitting diseases. The use of medicinal plants is considered an alternative to the recurring resistance to chemicals. Due to the need for efficient alternatives with less environmental impact, this study aimed to develop contact formulations with essential oils from the Java citronella (Cymbopogon winterianus) and clove (Syzygium aromaticum) plants and to assess in vitro the effects in different stages of the tick cycle. In the present study, concentrations from 0.5-15.0% of the essential oils incorporated in the formulations were used. The ticks from different geographical areas were treated with those formulations, and their effects on the production levels of eggs, on the larvae hatching, and their efficiency on ticks were assessed. The obtained results were compared with other commercial acaricidal products. After the 20th day of treatment, the formulations with citronella essential oil had 2.09-55.51% efficiency, depending on the concentration of the oil incorporated. The efficiency of the treatment with formulations containing clove essential oil was higher, from 92.47-100%. The results showed the acaricidal effects of the formulations tested when compared to commercial chemical products. In vivo studies should be performed in order to assess the efficiency of those formulations in the fields, aiming to use these products as an alternative for controlling cattle ticks.

Thymol is a monoterpene with proven acaricide action for several tick species. In addition to killing these ectoparasites, thymol can also reduce oviposition and egg hatch rate. However, the effects of thymol on the morphophysiology of tick ovaries are still unknown. Thus, the aim of this study was to evaluate the morphophysiological changes caused by this active principle in ovaries of Rhipicephalus sanguineus after a 6-day feeding period, through the application of morphohistochemical techniques. After the feeding period, a total of 50 females were divided into five groups and immersed in the following solutions: (I) distilled water (control), (II) 30% ethanol (control), (III) 1.25 mg/mL thymol, (IV) 2.5 mg/mL thymol, and (V) 5.0 mg/mL thymol. The experimental groups were kept in a climatic chamber (27 ± 1 °C; RH 80 ± 10%) for 5 days. After this period, morphological (hematoxylin/eosin) and histochemical (von Kossa) techniques were applied after remotion of the ovaries. The morphological results revealed large vacuoles in germ cells at different developmental stages and invaginations that represent deformations in the chorionic membrane. From the results obtained in this study, it was concluded that thymol interfered with the development of oocytes, which showed degeneration signs. The treatment containing 5.0 mg/mL thymol affected more accentuately the morphological development. Moreover, thymol also altered the calcium content of yolk granules, which generally showed an intense staining for this element.

The aims of this work were to identify the compounds and to investigate the acaricidal activity of the essential oil of Lippia sidoides for unengorged larvae and nymphs of Rhipicephalus sanguineus and Amblyomma cajennense. The oil was analyzed by gas chromatography and gas chromatography/mass spectrometry. In total, 22 compounds comprising 98.5% of the total peak area were identified. The major constituent of the essential oil was thymol (69.9%). The acaricidal activity against larvae and nymphs was assessed using a modified larval packet test. In all experiments, oils were tested at concentrations of 2.35, 4.70, 9.40 14.10 and 18.80 mg/mL. The mortalities of larvae and nymphs of R. sanguineus were 20.6, 47.8, 73.6, 99.5 and 99.0% and 12.0, 50.0, 76.3, 96.0 and 96.1%, respectively. For larvae and nymphs of A. cajennense the rates of mortality were 41.9, 63.3, 77.8, 82.5 and 100.0% and 0.0, 32.8, 64.8, 71.1 and 94.0%, respectively. The LC 90 values of the L. sidoides oil were 11.56 and 12.97 mg/mL for larvae and nymphs of R. sanguineus and 15.70 and 18.52 mg/mL for larvae and nymphs of A. cajennense, respectively. The essential oil from L. sidoides has acaricidal activity on unengorged larvae and nymphs of R. sanguineus and A. cajennense., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The present study evaluated efficacy of thymol against engorged females and unengorged larvae of Rhipicephalus microplus when diluted in 1 % dimethyl sulfoxide (DMSO) (formulation 1) and diluted in hydro-ethanol solutions of 50 % (formulation 2), 40 % (formulation 3), and 30 % (formulation 4). The immersion test was used on the engorged females for all dilution methods, while the larval packet method was applied to the larvae to test formulations 3 and 4 since the other two had already been tested before. Two control groups were established in both experiments; in one, ticks were treated with 1 % DMSO, and in the other, with 50 % ethanol, with ten repetitions for each treatment. The groups were maintained at 27±1 °C and RH>80 ± 10 %. For engorged females, the parameters weight before oviposition, egg mass weight, hatching percentage of larvae, and percentage of control were evaluated, while for larvae, percentage of mortality was evaluated. The egg mass weight of all treatments differed significantly (p<0.05) from the control groups; however, the reduction in egg mass weight was more accentuated (p<0.05) in the treatments with formulations 1, 3, and 4. The values of hatching percentage of the group treated with formulation 2 and control group were statistically similar (p>0.05). The group treated with formulation 2 attained the lowest percentage of control (66 %). In the other treatments, the control percentages were greater than 99 %. In larval test, the mortality was 100 % in all the treatments, and there was no mortality in the control groups.

In vitro effect of the association of the entomopathogenic nematodes Heterorhabditis bacteriophora HP88 and Heterorhabditis indica LPP1 with the essential oil of Lippia sidoides on engorged females of Rhipicephalus microplus was assessed. Engorged females with homogeneous weights (p >0.05) were divided into six groups of ten ticks each (each female = an experimental unit). In the treated groups, the ticks were exposed to the oil (40 μl/ml=4%) and nematodes (300 entomopathogenic nematodes (EPNs/tick) separately and also to the nematodes together with the oil, while in the control group, the females were immersed in Tween 80 (3%). All the treatments caused a significant reduction in the egg mass weight (p <0.05). The hatching percentage of larvae was reduced (p <0.05) in the groups treated with H. bacteriophora HP88, H. indica LPP1, and H. bacteriophora HP88 associated with the oil, but in the group treated only with the oil, the hatching of percentage of larvae rate was similar to that of the control group (p >0.05). The percentage of control in the group exposed only to the oil was 50.7%, while in the other treated groups, the percentage of control was greater than 95 %, reaching 100 %in the group treated with H. indica LPP1 associated with the oil. The results of the L. sidoides essential oil in the concentrations tested in association with both H. bacteriophora HP88 and H. indica LPP1 indicate that these combinations can be used to control R. microplus.

The objective of this study was to assess the acaricidal activity of carvacrol, thymol, eugenol, and (E)-cinnamaldehyde on unengorged larvae and nymphs of Amblyomma cajennense and Rhipicephalus sanguineus, using the modified larval packet test. Carvacrol, eugenol, and (E)-cinnamaldehyde were tested at concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 μl/ml, while thymol was tested at concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 mg/ml, in all cases with 10 repetitions per treatment. For the A. cajennense larvae, mortality rates caused by carvacrol, thymol, eugenol, and (E)-cinnamaldehyde at the lowest concentration were 45.0, 62.7, 10.2, and 81.6%, respectively, reached 100% at the concentration of 5.0 μl/ml for carvacrol and (E)-cinnamaldehyde and 5.0 mg/ml for thymol, while this mortality was observed at 15.0 μl/ml for eugenol. For the nymphs of this species, carvacrol and thymol caused 100% mortality starting at a concentration of 5.0 μl/ml and 10.0 mg/ml, respectively, while eugenol caused 100% mortality at 20.0 μl/ml and the mortality caused by (E)-cinnamaldehyde did not exceed 64%. In the tests with R. sanguineus larvae, the lowest concentration of carvacrol and (E)-cinnamaldehyde resulted in 100% mortality, while this percentage was observed starting at 10.0 μl/ml for eugenol. For nymphs, carvacrol and thymol at the smallest concentration caused 100% lethality, unlike the results for eugenol and (E)-cinnamaldehyde, where 100% mortality was only observed starting at the concentration of 10.0 μl/ml. The results obtained indicate that the tested substances have acaricidal activity on unengorged larvae and nymphs of A. cajennense and R. sanguineus.

The monitoring of resistance of cattle tick populations in Brazil to the chemical bases in use is largely limited to investigation of the phenotypic profile. There are few studies investigating the role played by the genotypic profile in acaricide resistance in the country. Therefore, the aim of the present study was to carry out molecular characterization and trace out the genetic profile of populations of Rhipicephalus microplus with respect to resistance to the organophosphate and pyrethroid chemical groups. For that purpose, larvae were genotyped belonging to 587 populations for pyrethroids and 306 for organophosphates, using the polymerase chain reaction technique. It was found that 75.49% and 97.44% of the larvae studied showed resistance to the organophosphates and pyrethroids, respectively. Among the populations resistant to pyrethroids, 91.9% were heterozygotes, showing that most of the resistant populations have only one allele responsible for resistance. Therefore, it is possible to conclude that the genotyped populations have high resistance to organophosphates, and even more so to pyrethroids. This information is fundamental for understanding the mechanisms of resistance of R. microplus to acaricides, to enable improvement of control techniques., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The repellent activity of eugenol was evaluated on Rhipicephalus microplus and Dermacentor nitens larvae in a bioassay based on the vertical migration of these tick larvae during the host-seek phase. Solutions were used at concentrations of 10, 20, 30, 40, and 50 μl/ml and a control only with the solvent ethanol 50 GL. The larvae's reaction of climbing the rods of filter paper was observed each hour for 12 h and once more at 24 h. The repellence was calculated as a percentage through visual estimation, from the tendency of the larvae to climb the rods when the top part was impregnated with the eugenol solutions. For R. microplus larvae, the four highest concentrations produced repellence levels were greater than 80 % for up to 9 h. The repellence rates after 10 h for the concentration of 10 μl/ml was lower than 2 % (p > 0.05); and after 24 h, the values observed for the concentrations of 40 and 50 μl/ml were 77 and 80 % (p < 0.05). For the D. nitens larvae, the repellence was greater than 80 % for up to 5 h at the concentrations of 40 and 50 μl/ml; and after 9 h, the repellence fell for all treated groups were significantly different than the control were found only at the three highest concentrations. Therefore, eugenol has repellent activity on the larval stage of these two tick species, with the R. microplus larvae being more sensitive.

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.

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.

The aim of this work was to identify the compounds and to investigate the acaricidal activity of the essential oil from the leaves of Lippia sidoides on Rhipicephalus microplus and Dermacentor nitens. The oil was obtained by hydrodistillation and analyzed by gas chromatography (GC/FID) and gas chromatography/mass spectrometry. In total, 15 compounds comprising 99.97 % of the total peak area were identified. The main constituent of the essential oil was thymol (67.60 %). The acaricidal activity was assessed by the modified larval packet test, with oil concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 μl/ml, and by the female immersion test with concentrations of 10.0, 20.0, 40.0, 60.0, and 80.0 μl/ml. The mortality of the R. microplus and D. nitens larvae was greater than 95 % starting at concentrations of 10.0 and 20.0 μl/ml, respectively. In the test with the engorged females, the L. sidoides essential oil starting at a concentration of 40.0 μl/ml caused a significant reduction (p < 0.05) in the values of the egg mass weight and egg production index. The viability of the eggs was affected in all the treated groups, with significantly lower hatching rates (p < 0.05) in relation to the control group. The control percentages at concentrations of 10.0, 20.0, and 30.0 μl/ml were 54, 57, and 72 %, and reached 100 % at the highest two concentrations (60.0 and 80.0 μl/ml). Therefore, it can be concluded that the essential oil from the leaves of L. sidoides has acaricidal activity on R. microplus and D. nitens.

The acaricidal activity of three thymol formulations was investigated at five concentrations (1.25, 2.5, 5.0, 7.5, and 10.0 mg/ml) on Rhipicephalus microplus larvae, and the behavior of its solubility in these formulations was analyzed. The thymol was dissolved in distilled water plus 1 % dimethylsulfoxide as adjuvant under two heating regimes (water bath in formulation 1 and hot plate in formulation 2) as well as without heating in 50 % ethanol and 50 % water (v/v). The acaricidal activity was assessed by the modified larval packet test, and the solubilization behavior was investigated by ultraviolet-visible spectroscopy, based on the Beer-Lambert law. With formulations 1 and 2, the mortality was greater than 95 % starting at the thymol concentrations of 5.0 and 7.5 mg/ml, respectively, while with formulation 3, this mortality level was reached starting at a concentration of 2.5 mg/ml, showing that the addition of ethanol in the solution enhanced the acaricidal action of thymol. This result was supported by the LC 90 values, which were 3.3, 2.4, and 1.6 mg/ml of thymol for formulations 1, 2, and 3, respectively. This result is related to the better solubility of thymol in the hydroethanolic formulation, since the spectroscopic analysis revealed that the thymol dissolved more completely in this formulation. This fact was evident once the R (2) obtained from the linear regression analysis of the relation absorbance × concentration of the formulations 1, 2, and 3 approached the optimal value (R (2) = 1) in the following sequence: 1, 2, and 3 (0.717, 0.901, and 0.968, respectively).

The aim of this study was to evaluate the acaricidal activity of eugenol, with different solubilizations and concentrations, on Rhipicephalus microplus and Dermacentor nitens larvae and to determine the lethal time. The study consisted of four experiments, and the mortality was assessed using the larval packet test with adaptations. The mortality observed in the first experiment was 100 % for all the groups treated with eugenol solubilized in different solvents. In the second, the hydroethanolic formulation of eugenol was used, and the mortality rates for R. microplus and D. nitens was 100 % starting from the concentration of 5.0 μl/ml. In the third experiment, the mortality was 100 % for larvae of both R. microplus and D. nitens after 1 h of contact. And in the fourth experiment, the mortality was above 90 % and statistically similar (p > 0.05) for the four methods the test evaluated.

The objective of this work was to verify the sensitivity of Amblyomma cajennense and Dermacentor nitens larvae to the solvents ethanol, methanol, acetone, xylol and dimethyl sulfoxide (DMSO) and the surfactant Tween 80. The first four solvents were tested at analytical purity while the DMSO and surfactant Tween 80 were tested at a concentration of 1%. The substances tested at analytical purity that caused high mortality were also tested at concentrations of 50%, 25% and 1%. The larval packet test was used, with 10 repetitions for each treatment. A control group was also formed with the same number of repetitions, in which the larvae were only exposed to distilled water. In the first experiment, only xylol was highly toxic at the concentrations tested, causing mortality above 90% for larvae of both species. In the second experiment, xylol at 1% and at 25% showed low toxicity to the A. cajennense and D. nitens larvae, respectively, since the percentage mortality was statistically similar to that of the control group (p>0.05)., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The aim of this study was to assess the acaricidal activity of hydroethanolic formulations of thymol at varying concentrations on Rhipicephalus sanguineus and Dermacentor nitens larvae. The larval packet test was used and the thymol concentrations tested were 2.5, 5.0, 10.0, 15.0 and 20.0 mg/ml. The control group was exposed only to water and ethanol (50/50%) and there were 10 repetitions for each treatment. The mortality was evaluated after 24 h. For the R. sanguineus larvae, the mortality rates were 47.5, 50.2, 96.7, 95.9 and 98.1%, while for D. nitens the rates were 14.1, 75.0, 90.2, 90.3 and 99.5%, at respective thymol concentrations of 2.5, 5.0, 10.0, 15.0 and 20.0 mg/ml. The results indicate that the hydroethanolic formulations of thymol tested have acaricidal activity on R. sanguineus and D. nitens larvae exposed topically, causing mortality greater than 90% 24 h post-treatment starting at the concentration of 10 mg/ml., (Copyright © 2011 Elsevier B.V. All rights reserved.)

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.

With the objective of encouraging the use of rabbits as alternative hosts for the cattle tick, four rabbits were infested on the ears and back. From the second day of infestation the developmental stages were observed. e duration of larval engorgement and changes were estimated between six and eight days in the region of the back and between 've and seven days in the pinna. e nymphal engorgement and changes occurred at approximately 10.80 ± 2.65 days in the dorsal and 11.00 ± 2.52 days in the ear, with the engorgement of adults and copulation occurring at 7.03 ± 2.45 days, on the dorsal region and 8.55 ± 1.82 days in the region of the pinna. e parasitic period ranged from 21 to 29 days (back) and from 23 to 30 days (pinna). e engorged females of the back and ear weighed on average 34.43 ± 18.73 and 36.30 ± 18.10 mg, respectively. e nutritional and reproductive efficiency indexes were 17.38 ± 14.27 and 26.85 ± 17.13% (back) and 17.42 ± 12.22% and 30.70 ± 19.80% (pinna). Although not appropriate to maintain successive generations of Rhipicephalus (Boophilus) microplus, rabbits can be used for different stages of engorgement of the ixodid, allowing experimental studies.

The aim of the present study was to evaluate the acaricidal efficacy of thymol on unengorged and engorged larvae and engorged nymphs of Amblyomma cajennense. To perform the test for unengorged larvae, the larval packet technique was employed and mortality was evaluated 24h after the test, while for engorged larvae and engorged nymphs the immersion technique was employed and mortality was evaluated after 15 days. In all the experiments, the following concentrations of thymol were tested: 2.5, 5.0, 10.0, 15.0 and 20.0mg the thymol/ml. The control group was exposed to water and 1% DMSO and there were 10 repetitions for each treatment. The values found for mortality of unengorged larvae were 18.2%, 51.8%, 97.6%, 93.5% and 94.5%, and for engorged nymphs the values were 26.0%, 92.2%, 100.0%, 100.0%, 100.0%, in concentrations of 2.5, 5.0, 10.0, 15.0 and 20.0mg of thymol/ml, respectively. In the test with engorged larvae, mortality was 100.0% in all treatments. Based on the results, it is possible to conclude that thymol has acaricidal activity against immature stages of A. cajennense., (Copyright © 2011 Elsevier B.V. All rights reserved.)

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.

The present work had the objective of evaluating the influence of different concentrations of thymol on the biological parameters of engorged females of Rhipicephalus (Boophilus) microplus and also its ovicide activity on eggs of this tick. In order to carry out the work, four groups were formed, each containing 20 engorged females, which were immersed for 5 min in different concentrations of thymol (1.0%, 1.5%, and 2.0%) and a control group (water + dimethylsulfoxide). The following biological parameters were observed: initial weight (mg); egg mass weight (mg); pre-oviposition, oviposition, and survival period (days); hatching percentage (%HP); egg production (%EPI) and nutritional (%NI) indexes; and the percentage of control (%C). In order to perform the second stage, thymol solutions were sprayed on the egg masses (50 mg). The parameters' initial weight and pre-oviposition did not present significant differences (p>0.05) among the groups. Thymol caused alterations in the parameters egg mass weight, oviposition, survival period, hatching percentage, EPI, and NI, presenting highly significant differences between the treatments and the control group (p<0.01). The control efficacy was higher than 95% in all the treatments, reaching 99% in the concentration of 2.0%. In the ovicide test, the hatching percentage was not affected in any of the treatments (p>0.05). In the second experiment, when eggs were treated directly, the thymol did not affect significantly this parameter. These results demonstrate that thymol showed deleterious action on most of the analyzed parameters; thus, it is possible to conclude that, in laboratorial conditions, this monoterpene was efficient in the control of engorged females of R. (B.) microplus.

The present work was aimed at evaluating the "in vitro" efficacy of different concentrations of thymol on engorged nymphs and females of Rhipicephalus sanguineus. The nymphs were separated in seven groups and immersed in different concentrations of thymol (0.25%, 0.5%, 1.0%, 1.5%, and 2.0%) for 5 min. A control group was established (water + dimethylsulfoxide) together with a positive control group (Amitraz*), and mortality was evaluated after 15 days. In order to perform the experiment with females, the same methodology was used, and the following concentrations were tested: 1.0%, 1.5% and 2.0%. After 5 min of immersion, the engorged females were fixed on Petri dishes with the aid of a double-sided adhesive tape and kept in a climatized room regulated at 27 +/- 1 degrees C and UR >80 +/- 10%, and different parameters referring to the biology of the non-parasitary phase were evaluated daily. The values found for thymol efficacy on nymphs were 0.0%, 100%, 100%, 100%, and 100% in concentrations of 0.25%, 0.5%, 1.0%, 1.5%, and 2.0%, respectively. In the experiment with engorged females, thymol did not induce any significant alterations (p < 0.05) in the parameters of weight alteration, egg mass weight, pre-oviposition period, hatching percentage, egg production index and nutritional index; however, it affected the engorged females final weight in all treatments (p > 0.05). The concentration of thymol 2% was the one that showed a better efficacy (41%). It was concluded that thymol had a more accentuated deleterious effect on engorged nymphs, and it might be a promising supporting agent for the control of this ixodid.

This study evaluated aspects of population ecology and community component of mites in Pseudolynchia canariensis (Macquart) of domestic pigeons Columba livia in Juiz de Fora, Minas Gerais, Brazil. A total of 236 forest mites were found in 74 out of 156 fl ies (47.4% infestation). The prevalence for Myialges anchora Sergent & Trouessart was 23.7%, with average intensity 1.5 +/- 0.90 and average abundance 0.4 +/- 0.76, mostly (94.4%) located on the abdomen; 75.9% of the mites (n = 41) surrounded by eggs, with a mean of 16.4 +/- 14.60 eggs per individual and 21.6 +/- 12.90 eggs per ovigerous female. The prevalence for M.lophortyx (Furmann & Tharshis) was 13.5% (n = 21) with 62 individuals, mean intensity 3.0 +/- 2.75 average abundance 0.4 +/- 1.41; 41.9% (n = 26) located at the right wing and 58.1% (n = 36) were in the left wing; 72.6% (n = 45) showed ovigerous mass, with 4.56 +/- 3.42 per each individual and 6.3 +/- 3.04 per ovigerous female. The prevalence of Ornithocheyletia hallae Volgin was 23.7% (n = 37) with 120 individuals, with mean intensity of 3.2 +/- 4.47, and mean abundance of 0.8 +/- 2.56. In 28.4% (n = 21) of flies occurred simultaneous mite infestations, mainly for M.anchora and O.hallae, present in 66.7% (n = 14) of flies with mixed effect. The component communities showed an aggregate distribution. The mite species O.hallae was more agregate, while M.anchora and M.lophortyx showed the same K value.

The present work was aimed at evaluating the in vitro efficacy of thymol on engorged and unengorged larvae of Rhipicephalus sanguineus. In order to perform the study, unengorged larvae were interleaved on filter paper (2 x 2 cm) recently sprayed with thymol in the following concentrations: 0.25%, 0.5%, 1.0%, 1.5%, and 2.0%. For each concentration, ten repetitions of 50 larvae each were performed. A control group was established (water + DMSO) together with a positive control group (Amitraz), with ten repetitions each. Mortality was evaluated after 24 h. For the test with engorged larvae, the same thymol concentrations were used with the larvae being immersed for 5 min and mortality evaluated after 15 days. The values found for thymol efficacy for unengorged larvae were of 21.3%, 17.4%, 19.1%, 32.1%, 37.7% and for engorged larvae 3.0%, 0.0%, 3.0%, 97.0%, and 100.0% in concentrations of 0.25%, 0.5%, 1.0%, 1.5%, and 2.0% respectively. We have concluded that thymol had a deleterious effect on larvae of R. sanguineus, especially on engorged larvae for which 100% mortality was found in higher concentration.

This study was aimed at making the bionomic aspects of Pseudolynchia canariensis (Macquart 1839) clear and at providing ways for keeping this diptera under laboratory conditions. A hundred and seventy-four flies were collected, of which 30% did not show hematophagy under laboratory conditions. The others were split into two groups with feeding intervals of 24 and 48 h. The individuals fed within a 24-h interval were found to live longer than the ones fed within a 48-h interval and blood meal time was decreased as feeding interval was increased. In the 48-h-feeding-interval group, females were found to live longer than the males, what could not be shown for the 24-h-feeding-interval group. Copulation was not observed in vitro, even after exposure to pigeon feathers and scraped skin. Data from this study suggest that daily feeding makes it possible to keep P. canariensis under laboratory conditions for a period of time longer than the one found for the 48-h feeding interval.

The present work studied the characterization of ectoparasites on dogs in the nucleus of urban expansion of Juiz de Fora, Minas Gerais, Brazil. The collections were conducted from July to September 2003, examining 101 dogs of no defined breed. The ectoparasites were collected through visual and tactile inspection of the animals' bodies, and were then placed in bottles with 70 degrees GL ethanol and identified under a stereomicroscope. Specimens were clarified and mounted on slides for analysis under a photonic microscope. Among the siphonaptera, Ctenocephalides felis was the most prevalent (64.35%), with mean intensity of 6.12 +/- 5.37 Siphonaptera/dog, followed by Rhopalopsyllus lutzi (3.96%), a hybrid of C. felis X C. canis (1.98%), and Tunga penetrans (1.98%). The only species of Phthiraptera found was Trichodectes canis (7.92%), with 1.3 +/- 0.51 Phthiraptera/dog. Among the Ixodidae, Rhipicephalus sanguineus was the most prevalent species (49.50%), with a mean intensity of 6.44 +/- 10.2 Ixodidae/dog, followed by Amblyomma cajennense (3.96%), Boophilus microplus (2.97%), A. ovale (1%) and A. aureolatum (1%). The ixodid nymphs were separated into a Amblyomminae nymphs (58.41%), with 10.11 +/- 10.09 nymphs/dog, and Rhipicephalinae nymphs 1(24.75%), with 2.64 +/- 3.25 nymphs/dog. Ixodid larvae were found on 3.96 % of the dogs.

The objective of this work was to present a new technique for feeding Pseudolynchia canariensis, reducing both fly-host and researcher-host contact. Flies obtained from pigeons caught in Juiz de Fora, MG, Brazil were housed in screen top test tubes and kept at 27 degrees C 80%RH. Three infestation-free pigeons were kept in screen cages as their food source. Pigeons were restrained on a wooden frame with test tube supports and a muzzle and twine to immobilize the head and wings, respectively. To minimize bird stress, both feeding wing and birds were alternated. Using this new technique, 60 flies were kept under laboratory conditions for an average of 12 +/- 17.8 days. This method allowed up to six flies per time to be fed in a short space of time, thus helping reduce stress for the animals used.

During the study of the metazoan parasites of Paroaria dominicana Linnaeus, 1758 (Passeriformes), four infrapopulations of Platynosomum illiciens (Brown, 1901) (Trematoda), parasitic in the liver ducts were collected, with intensity of infection 10, 11, 18 and 36 specimens respectively. Differences between the means of morphometrical values and the total number of eggs were statistically tested. The liver weigh were not significantly correlated with avian total weigh and length, and with the total parasitic abundance. Positive correlations were observed between the size of infrapopulations and the total number of eggs, and between parasite body size and the eggs. size. Negative correlations were observed between parasitic intensity and liver weigh; between the size of infrapopulations and the parasite body size; between total number and the size of eggs and between parasite body size and total number of eggs. These results suggests that host with smaller liver had greater infrapopulations. These greater infrapopulations showed smaller parasite body size and higher size and number of eggs.

The tick Boophilus microplus is the principal species of ectoparasite that impairs dairy cattle productivity in Brazil. Its control is mainly by using synthetic chemical products during its parasitic phase. The purpose of this study is to evaluate the acaricide activity of four products of natural origin. Depending on solubility, tests were conducted with solutions in distilled water or emulsified in aqueous DMSO at 1% of the following products: thymol, menthol, methyl salicylate, and salicylic acid. Each of these was tested at three concentrations (0.25, 0.5, and 1.0%) with five repetitions. The "larval packet test" was performed on approximately 100 larvae of B. microplus at around 15 days of age. After applying the test substances, the envelopes were kept at 27 degrees C and UR >80% and opened after 24 h to count the living and dead larvae. Of the four products tested, only thymol caused significant (up to 100%) mortality of B. microplus larvae. The mortality caused by the other products varied from 0.52 to 9.76%. Hence, thymol can be considered a potential agent to control bovine ticks.

The aim of the experiment was to evaluate the influence of different aggregations on the molting process of the nidicoulus ticks, Haemaphysalis leporispalustris. Tick-naïve domestic rabbits (Oryctolagus cuniculus) were used as experimental hosts for the parasitic phase. All free-living phases were kept into 4.5 mL glass vials in incubators at 27+/-1 degrees C, 80+/-15% relative humidity and scotophase. The periods of pre-molt, molt, percentage of ecdisis and molting pattern of larvae and nymphs were studied. A total of 1560 engorged larvae and nymphs were divided in six groups of aggregations: 1, 5, 10, 20, 40 and 80 ticks with ten repetitions. The molting of nymphs and adults were observed daily. The percentage of ecdysis was the only parameter with no statistical difference among the groups (p>0.05). All others parameters were statisticaly different (p<0.05). The results suggested that highest aggregations accelerate the molting process of H. leporispalustris. We recommended the following aggregations: 4.4 ticks/ml for scientific research and less than 7.7 tick/ml for ticks maintenance.

The species Hepatozoon procyonis Richards, 1961 was described in Procyon lotor in the USA and then in other reports in the USA, while in Panama H. procyonis has been described in Procyon cancrivorus. The objective of this paper is to report the occurrence of this species in the Brazilian procionids P. cancrivorus and Nasua nausa and to describe the morphology and morphometrics of the gametocytes. The analysis was based on blood smears, stained with Giemsa, which were examined under a photonic microscope. The morphometry was done with an ocular micrometer. It was based on the morphological characteristics and morphometric data on the gametocyte. It can be concluded that the species of the genus Hepatozoon that occurs in Brazilian procionids is the same as that occurring in procionids in Central and North America.

Oviposition, reproductive and nutritional efficiency indices (REI and NEI), eclosion and larvae mortality of Haemaphysalis leporispalustris were studied under controlled conditions, inside desiccators, at constant temperatures of 25+/-1 degrees C and relative humidity (RH) levels adjusted at 30, 50, 70 and 95%. Fourth generation tick specimens, originated from a colony fed on rabbits (Oryctolagus cuniculus), were used. The experiment was conducted between Jul/1999 and Jan/2001. The pre-oviposition and oviposition periods decreased and increased, respectively, according to the increase in RH. The egg mass weight, REI and NEI, increased according to the increase in RH. The females' residual weight, under high humidity levels, was probably small due to a better energetic use for egg production (51.77 +/- 11.06 mg at 95% RH). The incubation period was inversely proportional to RH increase (45.10+/- 2.47 days; 40.33+/- 1.45 days and 36.85+/- 0.37 days at 50% 70% and 95% RH, respectively). There was no eclosion at 30% RH. Period and percentage of eclosion were directly proportional to the increase of the other RH levels. A diminished formation of embryonic points as well as the desiccation of the already formed eggs were observed. Larval longevity was negatively influenced by 50% RH, with death occurring concomitant to eclosion.

Fleas and ticks transmit various pathogens while feeding on the blood of dogs. This study sought to verify the occurrence of ectoparasitism and hemoparasitism in dogs from two urban areas in the city of Juiz de Fora, Minas Gerais, Brazil. Between February and August 2003, 101 dogs were studied: 50 came from apartments in the downtown region and 51 from houses with grassy yards. The ectoparasites were collected and conserved in etanol 70%. The occurrence of hemoparasites was verified by examining blood smears from sample taken from the dogs'ears. The blood smears were stained with Giemsa and 100 fields per slide were examined, studying the erythrocytes to determine parasitism. From among the dogs living in apartments, we found (with respective prevalence and mean intensity): Ctenocephalides felis (12%), (3.3+/-2.0); Rhipicephalus sanguineus (2%); and ixodid nymphs (2%). In this environment in the dogs were not found hemoparasites. From the houses with grassy yards, we observed the following prevalence levels and mean intensities: C. felis (14%), (2.28+/-1.9); R. sanguineus (35%), (7.8+/-9.8); ixodid nymph (18%), (1.4+/-0.7); and ixodid larvae (4%), (12+/-14.4). The hemoparasites found were: Ehrlichia canis (16%) and Babesia canis (2%).

To evaluate the effect of the immersion of engorged larvae and nymphs of Amblyomma cajennense, these stages were immersed by 24, 48, 72 and 96 hours in test tubes containing distilled water. The period of premolt of the nymphs increased proportionally to the time of immersion (p<0.05). The ecdisis period, the longevity and the mortality (days) were similar in all of treatments (p>0.05). The percentage of ecdisis in nymphs was reduced after 72 hours of immersion of the larvae (p<0.05). None of the engorged nymphs survived 96 hours of immersion. The period of the adult premolt increased after 48 hours of immersion (p<0.05). The ecdisis period was similar in the treatments of 24 and 48 hours of immersion (p>0.05). The percentage of ecdisis decreased starting from 24 hours of immersion (p<0.05). There was not statistical difference (p<0.05) between the control group and the 24 hours group of immersion for the parameters longevity and mortality. These findings suggest that the engorged nymphs were more sensitive to the immersion that the engorged larvae.