Liver and spleen of hosts of Rhipicephalus linnaei exposed to synthetic (afoxolaner) and natural acaricides (esters from castor oil). A comparative clinical-morphological study.

In dogs, Rhipicephalus linnaei transmits pathogens such as Ehrlichia canis, Babesia vogeli, and Hepatozoon canis. The veterinary market has synthetic acaricides to ticks control. Esters derived from castor oil are efficient. However, there is little information about their effects on non-target organisms. This work consisted of a clinical (AST, ALT, and ALP) and histological and histochemical analysis (liver and spleen) of female rabbits exposed to these esters and afoxolaner. The rabbits were divided into three groups: control group (CG) received Bandeirante® rabbit feed; the afoxolaner treatment (TG1) received rabbit feed and two doses of afoxolaner; castor oil esters treatment (TG2) received rabbit feed enriched with esters (1.75 g esters/kg). No alterations were observed in the AST, ALT, and ALP enzymes in exposure to esters TG2. Rabbits from TG1 showed changes in AST. The liver of rabbits exposed to afoxolaner underwent histological and histochemical changes, such as steatosis and vacuolation, as well as poor protein labeling. Polysaccharides were intensely observed in the group exposed to esters. The spleen showed no changes in any of the exposure. Esters from castor oil caused fewer liver changes when incorporated into the feed and fed to rabbits than exposure to afoxolaner.

Exposure of Rhipicephalus sanguineus sensu lato Latreille, 1806 (Acari: Ixodidae) to hexane extract of Acmella oleracea (Jambu): semi-engorged and engorged ticks.

The present study analyzed the efficacy of hexane extract of Acmella oleracea against Rhipicephalus sanguineus sensu lato. After exposure of R. sanguineus s. l. female ticks to 14 different concentrations of the hexane extract of A. oleracea, the LC50 was established as 18.135 mg/mL (limits: 16.251-22.675). The effects of the extract were more significant in the ovary of the semi-engorged females, that presented damages in most oocytes. Since such changes would not be repaired, the oocytes could not advance to further stages of development (I-V), thereby causing the inhibition of ovary development, interruption of vitellogenesis, oocyte death and, consequently, infertility. The semi-engorged females also presented extensive damages in the midgut cells, which would prevent (totally or partially) these cells from functioning properly (blood intake and release of nutrients), impairing the ectoparasite nutrition and the viability of the individual. Thus, the hexane extract of A. oleracea can be considered as a promising alternative to chemical control of pests of medical and veterinary importance, due to its efficient acaricidal activity and lesser environmental impact, when used against female ticks in the middle feeding stage.

Rhipicephalus sanguineus salivary gland extract as a source of immunomodulatory molecules.

Rhipicephalus sanguineus sensu lato (s.l.), popularly known as 'brown dog tick', is the primary vector of pathogens affecting dogs worldwide. To enter the host's organism, these pathogens utilise the anticoagulant, antiplatelet, anti-inflammatory and immunomodulatory actions of compounds present in the tick's saliva; such compounds are released by the ectoparasite in order to attach and feed on dogs. Nitric oxide (NO) is one of the regulatory factors in inflammation, apoptosis and immunomodulation. Here, we evaluated the in vitro activity of salivary gland extract of female dog ticks on the macrophage-derived J774 cell line, with and without lipopolysaccharide (LPS) stimulation. Cultures were evaluated for possible morphological alterations caused by exposure to the extract. There was no apparent in vitro cytotoxicity of the extract. Also, the NO secretory response in the non-LPS-stimulated cells was not inhibited. On the other hand, the extract presented modulatory action in the cultures of LPS-stimulated cells at a concentration of 0.1 µg/mL, possibly through macrophage activation, and induced a significant decrease in NO secretion. These results confirm the modulatory potential of bioactive molecules in the salivary glands of R. sanguineus ticks.

Ultrasound-assisted extraction extracts from Stemona japonica (Blume) Miq. and Cnidium monnieri (L.) Cuss. could be used as potential Rhipicephalus sanguineus control agents.

Nicotiana tabacum, Stemona japonica, and Cnidium monnieri are common plants that are widely used for their anti-parasitic properties. The purpose of this study was to evaluate the acaricidal activity of extracts from these plants against the brown dog tick, Rhipicephalus sanguineus. A composition analysis of crude extracts by GC-MS was conducted to discover compounds with acaricidal effects. The toxicity of extraction against the engorged nymphs of R. sanguineus was evaluated by an immersion test. The results showed that the crude extracts of S. japonica and C. monnieri in varying ratios, concentrations, and from different extraction methods, had a killing effect on R. sanguineus. Lethality reached 76.67% ± 0.04410 when using a 1:1 extract of S. japonica:C. monnieri in 75% ethanol with ultrasonic extraction; the crude extract was determined at a concentration of 0.5 g/mL. GC-MS results showed that osthole and 5-hydroxymethylfurfural (5-HMF) are the main components of the extract. These results suggested that ultrasound-assisted extraction (UAE) extracts contained acaricidal components acting against R. sanguineus, which may result in the development of effective extracts of S. japonica and C. monnieri as a source of low-toxicity, plant-based, natural acaricidal drugs.

A comparison of the efficacy of two commercial acaricides (fipronil and amitraz) with Azadirachta indica (neem) on the brown dog tick (Rhipicephalus sanguineus) from canines in Trinidad.

The brown dog tick (Rhipicephalus sanguineus) is prevalent on canids in Trinidad. It is directly (by causing anaemia) and indirectly (by acting as a vector of tick-borne pathogens) responsible for morbidity and mortalities in the canine population. The most commonly used commercial acaricides available to pet owners in Trinidad are amitraz and fipronil. Often, these acaricides may be abused and misused in a desperate attempt to rid pets of ticks. The objective of this study was to compare the efficacy of amitraz and fipronil with the herbal alternative, neem (Azadirachta indica). Triplicate in vitro trials utilizing the Larval Packet Test (LPT) were conducted using three concentrations (low, recommended and high) of fipronil (0.025%, 0.05% and 0.1%), amitraz (0.01%, 0.02% and 1%), neem oil (10%, 20% and 40%) and neem leaf extract (0.25%, 0.5% and 2%) for each trial. Statistical analysis using the mixed-effect Poisson regression analysis indicated that there was a significant difference (p < .05) in the survival of ticks pre-treatment versus post-treatment with amitraz, fipronil and all controls when compared to the neem oil. Fipronil and amitraz caused ≥99% mortality for all concentrations used in this study. Mortalities for neem oil and neem leaf extract ranged from 72.7% to 82% and 38% to 95.3%, respectively, with the greatest percentage of mortalities occurring at the lower concentrations. Neem oil and neem leaf extract can be used as alternative acaricides, and however, they are less efficacious against the brown dog tick than amitraz and fipronil.

Efficacy of Tagetes minuta (Asteraceae) essential oil against Rhipicephalus sanguineus (Acari: Ixodidae) on infested dogs and in vitro.

Ticks from Rhipicephalus sanguineus complex are widely distributed in the world and one species from this complex is the most common tick on dogs in Brazil, notably in urban areas. This tick is a vector of several diseases. Among others it transmits the agent of canine Ehrlichiosis, a major dog infectious disease and the agent of Rocky Mountain spotted fever. This tick can spread rapidly and develop intolerable infestations within no time. Currently tick control is done with acaricides and demand for such drugs has grown fast. However, R. sanguineus has already developed resistance to the main active compounds and the development of new acaricides is necessary. Many essential oils of plants have acaricidal effect and may be an important source of molecules for the synthesis of new acaricide products. In this study, we evaluated the effectiveness of a new herbal phytotherapic, consisting of the essential oil of Tagetes minuta L., against R. sanguineus in vitro and on dogs undergoing experimental infestations. The product displayed 100% efficacy against larvae, nymphs and adults of the tick on all tested conditions.

Repellency to ticks (Acari: Ixodidae) of extracts of Nigella sativa (Ranunculaceae) and the anti-inflammatory DogsBestFriend™.

Motivated by observations that the canine anti-inflammatory cream DogsBestFriend™ (DBF) appeared to deter flies, mosquitoes, and ticks from treated animals, repellent efficacy bioassays using four species of ticks were conducted with three extracts of Nigella sativa L. (Ranunculaceae), a constituent of DBF. The DBF cream was tested against nymphs of lone star tick, Amblyomma americanum (L.). In vertical filter paper assays, the three extracts applied at 0.413 mg extract/cm(2) filter paper repelled 96.7-100 % of brown dog tick, Rhipicephalus sanguineus (Latreille) nymphs, whereas, at the same rate, only one extract repelled >90 % A. americanum nymphs. Adult (mixed sexes) American dog ticks, Dermacentor variabilis (Say), required a higher concentration to be repelled effectively; two extracts, applied at 0.827 mg extract/cm(2) filter paper, repelled ≥90 % of the D. variabilis. In contrast, all extracts applied at much lower concentration (0.206 mg extract/cm(2) filter paper) repelled 100 % adult blacklegged ticks, Ixodes scapularis Say (only females tested). Of the two more repellent extracts, one lost most of its activity against A. americanum nymphs in <4 h when applied at 0.827 mg extract/cm(2) filter paper, whereas the other repelled 66.7 % of the nymphs at 192 h after application. At 0.206 mg extract/cm(2) filter paper, one extract was as repellent as deet against A. americanum nymphs. In a vertical bioassay in which nylon organdy was substituted for filter paper, DBF, at the rates of 1.67 and 0.835 mg cream/cm(2), repelled 76.7 and 30.0 % A. americanum nymphs, respectively. These findings indicate that when applied appropriately DBF should afford some protection to canines against tick bites.

Tick repellents and acaricides of botanical origin: a green roadmap to control tick-borne diseases?

Arthropods are dangerous vectors of agents of deadly diseases, which may hit as epidemics or pandemics in the increasing world population of humans and animals. Among them, ticks transmit more pathogen species than any other group of blood-feeding arthropods worldwide. Thus, the effective and eco-friendly control of tick vectors in a constantly changing environment is a crucial challenge. A number of novel routes have been attempted to prevent and control tick-borne diseases, including the development of (i) vaccines against viruses vectored by ticks; (ii) pheromone-based control tools, with special reference to the "lure and kill" techniques; (iii) biological control programmes relying on ticks' natural enemies and pathogens; and (iv) the integrated pest management practices aimed at reducing tick interactions with livestock. However, the extensive employment of acaricides and tick repellents still remains the two most effective and ready-to-use strategies. Unfortunately, the first one is limited by the rapid development of resistance in ticks, as well as by serious environmental concerns. On the other hand, the exploitation of plants as sources of effective tick repellents is often promising. Here, we reviewed current knowledge concerning the effectiveness of plant extracts as acaricides or repellents against tick vectors of public health importance, with special reference to Ixodes ricinus, Ixodes persulcatus, Amblyomma cajennense, Haemaphysalis bispinosa, Haemaphysalis longicornis, Hyalomma anatolicum, Hyalomma marginatum rufipes, Rhipicephalus appendiculatus, Rhipicephalus (Boophilus) microplus, Rhipicephalus pulchellus, Rhipicephalus sanguineus and Rhipicephalus turanicus. Eighty-three plant species from 35 botanical families were selected. The most frequent botanical families exploited as sources of acaricides and repellents against ticks were Asteraceae (15 % of the selected studies), Fabaceae (9 %), Lamiaceae (10 %), Meliaceae (5 %), Solanaceae (6 %) and Verbenaceae (5 %). Regression equation analyses showed that the literature grew by approximately 20 % per year (period: 2005-2015). Lastly, in the final section, insights for future research are discussed. We focused on some caveats for future data collection and analysis. Current critical points mainly deal with (a) not uniform methods used, which prevent proper comparison of the results; (b) inaccurate tested concentrations, frequently 100 % concentration corresponded to the gross extract, where the exact amounts of extracted substances are unknown; and (c) not homogeneous size of tested tick instars and species. Overall, the knowledge summarized in this review may be helpful for comparative screening among extensive numbers of plant-borne preparations, in order to develop newer and safer tick control tools.

Effects of andiroba (Carapa guianensis) oil in ticks: Ultrastructural analysis of the synganglion of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae).

The present study performed an analysis of the ultrastructural changes induced by andiroba seed oil in the synganglion of Rhipicephalus sanguineus female ticks, aiming to provide scientific grounds to help in the creation of more specific and efficient methods of control. The synganglion consists of a mass of fused nerves externally covered by the neural lamella, a uniform and acellular layer. Just below, the perineurium is found, formed by glial cells. Internally, the synganglion is subdivided into an outer cortical region (cortex), which contains the cellular bodies of the neural cells and an inner region, the neuropile, formed by a set of nerve fibers (extensions of the neural cells). The results showed that the synganglion of females exposed to andiroba oil showed structural changes, such as: irregular and apparently thinner neural lamella, perineurium glial cells presenting large cytoplasmic vacuoles, decrease in the extensions of glial cells, separation of cortex cells, which were formerly attached through their membranes, neural cells presenting irregular plasma membranes and cytoplasm with autophagic vacuoles and mitochondria with disorganized cristae and in process of degeneration. This study confirmed the neurotoxic action of the andiroba oil, which would probably be able to impair the neural functions. Thus, it is suggested that this product has the potential to be used as an alternative method to control ticks.

Histopathological study of ovaries of Rhipicephalus sanguineus (Acari: Ixodidae) exposed to different thymol concentrations.

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.

Morphological alterations in the synganglion and integument of Rhipicephalus sanguineus ticks exposed to aqueous extracts of neem leaves (Azadirachta indica A. JUSS).

Currently, the necessity of controlling infestation by ticks, especially by Rhipicephalus sanguineus, has led researchers and public health managers around the world to search for new and more efficient control methods. This way, we can highlight neem (Azadirachta indica A. Juss) leaf, bark, and seed extracts, which have been very effective on tick control, and moreover causing less damage to the environment and to the host. This study showed the potential of neem as a control method for R. sanguineus through morphological and morphometric evaluation of the integument and synganglion of females, in semiengorged stage. To attain this, routine techniques of optical microscopy, scanning electron microscopy and morphometry of the cuticle and subcuticle of the integument were applied. Expressive morphological alterations were observed in both organs, presenting a dose-dependent effect. Integument epithelial cells and nerve cells of the synganglion showed signs of cell vacuolation, dilated intercellular boundaries, and cellular disorganization, alterations not previously reported in studies with neem. In addition, variations in subcuticle thickness were also observed. In general, the effects of neem are multiple, and affect the morphology and physiology of target animals in various ways. The results presented in this work are the first evidence of its effects in the coating and nervous system of ticks, thus allowing an indication of neem aqueous extracts as a potential control method of the brown dog tick and opening new perspectives on acaricide use.

Acaricidal activity of essential oil from Lippia sidoides on unengorged larvae and nymphs of Rhipicephalus sanguineus (Acari: Ixodidae) and Amblyomma cajennense (Acari: Ixodidae).

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.

Acaricidal activity of extract of Artemisia absinthium against Rhipicephalus sanguineus of dogs.

The objective of the study was to evaluate the in vitro efficacy of different concentrations of chloroform extract obtained from the aerial parts of Artemisia absinthium in comparison to amitraz on adults, eggs and larvae of the dog tick Rhipicephalus sanguineus using the adult immersion test (AIT), egg hatchability test (EHT) and larval packet test (LPT), respectively. Five concentrations of the extract (1.25, 2.5, 5, 10 and 20 %) with three replications for each concentration were used in all the bioassays. A control group was established (water + dimethylsulphoxide) together with a positive control group (amitraz), with three repetitions each. In AIT, the mortality rates were 0.0, 13.3, 16.7, 33.3 and 93.3 % in concentrations of 1.25, 2.5, 5, 10 and 20 %, respectively, and the variation was significant (p=0.0151). The LC50 (CI) and LC95 (CI) values were calculated as 8.793 % (8.217-9.408) and 34.59 % (29.71-40.26), respectively. The egg production was reduced by 6.6, 6.6, 18.3, 42.5 and 85.1 % in the concentrations of 1.25, 2.5, 5, 10 and 20 %, respectively, and it was statistically significant (p=0.0274). In EHT, hatching was completely inhibited at 5, 10 and 20 % displaying 100 % ovicidal action while at the concentrations of 1.25 and 2.5 %, the inhibition rates were 20 and 60 %, respectively. In LPT, the extract caused 100 % mortality of larvae in the concentrations of 5, 10 and 20 % after 24 h while at the concentrations of 1.25 and 2.5 %, the mortality rates were 54.3 and 96.7 %, respectively. The LC50 (CI) and LC95 (CI) values were determined to be 1.11 % (1.099-1.121) and 2.37 % (2.328-2.423), respectively. The results show that the extract of A. absinthium has acaricidal properties and could be useful in controlling R. sanguineus which is an efficient vector of pathogens both in dogs and humans.

Action of andiroba oil and permethrin on the central nervous and reproductive systems of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) ticks females. A confocal study.

Research for acaricides with lower toxicity and impact on the environment has been intensified. An alternative would be the use of natural compounds or of synthetic products in lower concentrations than the ones sold commercially. Thus, this study describes the action of andiroba seed oil on the nuclei of the ovary and synganglion cells of Rhipicephalus sanguineus, and presents an analysis of the nuclear morphology of the nervous system cells of this tick species when exposed to permethrin. The results obtained showed that, although no changes have been observed in the genetic material of the ovary cells exposed to the andiroba oil, this compound, as well as permethrin, has neurotoxic action on the females of this species. The damages caused to the physiology of the synganglion, due to the loss of integrity of the genetic material, would result in the impairment of the metabolism of other systems of R. sanguineus ticks.

Morphological and cytochemical changes in synganglion of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) female ticks from exposure of andiroba oil (Carapa guianensis).

Actually, the most used method to control ticks is synthetic acaricides with neurotoxic action. However, the use of these methods presents inconveniences, such as the contamination of the environment and risks to the host's health due to the residual effects. Thus, several studies have been developed aiming to find alternative ways to control these ectoparasites, such as the use of natural compounds with active ingredients, which act controlling some species of plagues in addition to presenting medicinal properties that are beneficial to humans. The present study aimed to analyze the action of andiroba oil (Carapa guianensis) on the synganglion of Rhipicephalus sanguineus semiengorged females through morphological and cytochemical techniques aiming to verify if this natural product have neurotoxic action as the numerous synthetic acaricides. The results showed that andiroba oil interferes in the synganglion through structural and enzymatic changes, which lead the nervous tissue to apoptotic death involving autophagy. Among these changes was observed the emergence of large empty spaces between the perineurium and the cortical region, vacuolated cortex cells and with cell swelling, neural cells with picnotic nuclei or in initial stage of chromatin margination and neuropile with high structural disorganization. Considering these data, it can be concluded that andiroba seed oil can be used as an alternative method in the control of R. sanguineus ticks due to its neurotoxic action.

Rhipicephalus sanguineus (Acari: Ixodidae) female ticks exposed to castor oil (Ricinus communis): an ultrastructural overview.

Tick control has been accomplished through the use of synthetic acaricides, which has created resistant individuals, as well as contaminating the environment and nontarget organisms. Substances of plant origin, such as oils and extracts of eucalyptus and neem leaves, have been researched as an alternative to replace the synthetic acaricides. Ricinoleic acid esters from castor oil have recently been shown as a promising alternative in eliminating bacterial contamination during ethanol fermentation, by acting as an effective biocide. The same positive results have been observed when these esters are added to the food given to tick-infested rabbits. This study tested the effect of these substance on the reproductive system of Rhipicephalus sanguineus females, added to rabbit food, more specifically on oogenesis. For this, four groups were established: four control groups (CG1, CG2, CG3, and CG4) and four treatment groups (TG1, TG2, TG3, and TG4) with one rabbit in each (New Zealand White), used as hosts. After full 4 days feeding (semi-engorgement), the females were collected and had their ovaries extracted. In this study, it was observed that R. sanguineus females exposed to esters had their ovaries modified, which was demonstrated through transmission electron microscopy techniques. The addition of ricinoleic esters to the diet of tick-infested rabbits revealed how toxic such substances are for the cytoplasmic organelles of oocytes and pedicel cells. These compounds can change the morphophysiology of germ and somatic cells, consequently influencing their viability and, therefore, confirming that the ricinoleic acid esters from castor oil are a promising substance in the control of R. sanguineus.

Effect of ricinoleic acid esters from castor oil (Ricinus communis) on the oocyte yolk components of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae).

Rhipicephalus sanguineus are bloodsucking ectoparasites, whose main host is the domestic dog, thus being present in urban areas and closely located to people. Eventually, this tick species parasitize humans and can become a potential vector of infectious diseases. Methods to control this type of pest have been the focus of many research groups worldwide. The use of natural products is increasingly considered nowadays, due to the low toxicity levels to the host and low waste generation to the environment. This study tested the effect of ricinoleic acid esters from castor oil (as an potential acaricide) on the reproductive system of R. sanguineus females, more specifically on the vitellogenesis process. For this, two groups were established: the control group (CG) and the treatment group (TG) with five rabbits in each (New Zealand White), used as hosts. NaCl and ester were added to rabbits' food and offered to the hosts. After full engorgement, the females were collected and had their ovaries extracted. The ticks ovaries were submitted to histochemical techniques so the effects of esters could be observed over polysaccharides, proteins and lipids yolk. Changes in the deposition of yolk components were observed. This caused modifications on elements of polysaccharide origin and on glycoprotein compounds, interfering in the final yolk synthesis and compromising the development of the future embryo.

Action of andiroba oil (Carapa guianensis) on Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged females: morphophysiological evaluation of reproductive system.

Because of the increasing medical-veterinary importance of ticks, the development of alternative control methods, less aggressive to the host and the environment has become the target of several researches. In this sense, the present study analyzed the action of different concentrations (5, 10, and 20%) of andiroba seed oil (Carapa guianensis) on the reproductive system of Rhipicephalus sanguineus females, through histochemical techniques and the quantification of the reproductive efficiency index. The results showed that andiroba oil is a potent natural agent, able to cause several changes in the oocytes of this species, impairing the reproductive success, once this natural product induces great physiological changes in the oocytes in all development stages, such as drastic reduction in proteins, polysaccharides, and lipids in these cells, and these components are essential for the viability of the embryo. In addition, it was observed that this product stimulate the oviposition, mainly at the concentration of 20%. This higher production of eggs represents a defense mechanism developed by the organism in order to ensure the reproductive success of the species, even in the presence of the toxic agent. However, the results obtained suggested that the laid eggs would not be viable, due to the great changes undergone by the oocytes. Thus, the present study showed that the use of this vegetal product would be an alternative way to control the ticks, bringing benefits similar to the ones obtained through the use of synthetic acaricides; however, with less damage to nontarget organisms and the environment as well.

Cytotoxic effects of andiroba oil (Carapa guianensis) in reproductive system of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged females.

The present study performed an analysis about the effects of andiroba seed oil (Carapa guianensis) in the ovary of Rhipicephalus sanguineus semi-engorged females; once, there are few studies about the action of natural products on the reproductive system, a vital organ for the biological success of this animal group. The results showed that andiroba oil is a potent natural agent which causes significant structural changes in the oocytes, such as the emergence of large vacuolated cytoplasmic regions, reduction in the number of yolk granules, changes in the shape of the cells, as well as impairment of genetic material. In addition, the ovary epithelium showed severe morphological changes, such as extreme structural disorganization, with highly vacuolated cells and picnotic nuclei, forming an amorphous mass. This study showed also that oocytes (mainly in the initial stages of development) and the ovary epithelium of R. sanguineus females subjected to different concentrations of andiroba oil presented morphological changes which became more numerous and intense as the concentration of the product increased. Based on the results, it can be inferred that although the defense mechanisms are developed by oocytes to recover the cellular integrity (presence of autophagic vacuoles), these cells are not able to revert the damage caused by this product. Thus, it can be concluded that although the damages caused to the oocytes by andiroba oil are comparatively less severe than the ones caused by synthetic acaricides, this product can be considered a potent natural agent that reduce and/or prevent the reproduction of R. sanguineus females, with the advantage of not causing environmental impact such as synthetic chemical acaricides.

Effect of Calea serrata Less. n-hexane extract on acetylcholinesterase of larvae ticks and brain Wistar rats.

Acetylcholinesterase (AChE), an enzyme that hydrolyses acetylcholine (ACh) at cholinergic synapses, is a target for pesticides and its inhibition by organophosphates leads to paralysis and death of arthropods. It has been demonstrated that the n-hexane extract of Calea serrata had acaricidal activity against larvae of Rhipicephalus (Boophilus) microplus and Rhipicephalus sanguineus. The aim of the present study was to understand the mechanism of the acaricidal action of C. serrata n-hexane extract are specifically to investigate the in vitro anticholinesterase activity on larvae of R. microplus and in brain structures of male Wistar rats. The n-hexane extract significantly inhibited in vitro acetylcholinesterase activity in R. microplus larvae and rat brain structures. The results confirm that inhibition of acetylcholinesterase is a possible mechanism of action of hexane extract at C. serrata.