Sodium dodecylbenzene sulphonate (SDBS) present in detergents: action on the gills, skin, and blood of D. rerio fish.

1. Sodium dodecylbenzene sulphonate (SDBS) is one of the surfactants used worldwide in detergents which, due to high residual discharges, has great potential to cause ecotoxicological impacts. Therefore, the sublethal effects of SDBS on the gills and skin of male Danio rerio fish were investigated.2. The fish were distributed into three groups: GC (control), GT1 (0.25 mg/L of SDBS), and GT2 (0.5 mg/L of SDBS) and exposed for 21 days. After the experiment, histopathological analyses of the gills, histochemical analyses (counting of mucous cells), and biochemical analyses (antioxidant defense enzyme analysis, SOD, and CAT) were conducted.3. A significant increase (p < 0.05) in the incidence of circulatory disorders, progressive, and regressive alterations occurred in the GT1 and GT2 groups. Due to these changes, the total histopathological index of the gills was higher in these groups. Mucous cells in the gills and skin increased. There was an increase in SOD activity and a reduction in CAT activity in these groups. Haematology revealed neutrophilia and lymphocytosis in the blood of GT1 and GT2.4. The results clearly demonstrate that a 21-day exposure to SDBS causes severe morphophysiological damage to the gills, skin, and blood of D. rerio fish.

Effects of sodium dodecylbenzene sulfonate (SDBS) on zebrafish (Danio rerio) gills and blood.

Sodium dodecylbenzene sulfonate (SDBS) is an important surfactant used as a cleaning agent and industrial additive to remove unwanted chemicals which have been detected in the aquatic environment. The aim of this study was to examine the toxicological potential of SDBS on the gills of adult male zebrafish (Danio rerio) exposed to this chemical. For the 96 hr acute exposure, fish were divided into three groups: control, 0.25 mg/L, and 0.5 mg/L of SDBS. After the experiment, morphophysiological analyses (gill histopathology and histochemistry), oxidative stress (determination of gill activities of superoxide dismutase (SOD) and catalase (CAT)), and hematological analyses (leukocyte differentiation) were conducted. Data demonstrated that SDBS at both tested concentrations altered the histopathological index and initiated circulatory disturbances, as well as adverse, progressive, and immunological changes in the gills. In the 0.5 mg/L group, SOD activity decreased significantly, but CAT activity was not altered. Prominent blood changes observed in this group were neutrophilia and lymphocytosis. The number of mucous and chloride cells increased significantly in both groups. Taken together, our findings demonstrated that exposure of D. rerio to SDBS, even for 96 hr, produced adverse morphological and hematological effects associated with a reduction in SOD activity. Our findings indicate that exposure of aquatic species to the anionic surfactant SDBS may lead to adverse consequences associated with oxidative stress. Therefore, this study highlights the risks that this substance may pose to aquatic ecosystems and emphasizes the need for further investigations and strict regulations on its disposal.

Effects of sodium dodecylbenzene sulfonate (SDBS), active principle of detergents, on the liver and kidney of zebrafish (Danio rerio).

Sodium dodecylbenzene sulfonate (SDBS), a predominant component in detergents, requires an evaluation of its toxicological potential due to its hazardous environmental levels. Therefore, we evaluated the toxicological effects of SDBS on the liver and kidney of male D. rerio. Therefore, we evaluated the toxicological effects of SDBS on the liver and kidney of male D. rerio. The fish were divided into three groups: 0.0 (control), 0.25, and 0.5 mg/L of SDBS with exposure for up to 96 hours. After exposure, histopathological, histochemical (hepatic glycogen content), and biochemical analyses (SOD and CAT enzyme analysis) were performed on both organs. The results showed significant histopathological effects, such as circulatory disturbances and progressive and regressive alterations, leading to an altered histopathological alteration index. SOD and CAT enzymes exhibited prominent changes. Thus, it became clear that the surfactant SDBS can cause serious hepatic and renal problems in D. rerio fish, even with short-term exposure, necessitating more stringent control and regulation in the disposal of this surfactant.

Efficacy of essential oils of Egletes viscosa and Lippia schaueriana on the reproductive biology of Rhipicephalus sanguineus sensu lato engorged females.

The study analyzed the chemical composition and the acaricide effect of Egletes viscosa Less (macela-da-terra) and Lippia schaueriana Mart. (lipia-da-serra) essential oils (EOs) on Rhipicephalus sanguineus s. l. (Acari: Ixodidae) engorged females. The chemical analysis (GC-MS and GC-FID) identified 27 components in E. viscosa EO and 18 in L. schaueriana EO, which comprise more than 98% of its constituents. The effects of the oils on the reproductive biology of R. sanguineus ticks were assessed by adult immersion test. Both EOs significantly reduced (p < 0.05) the egg production index when the females were exposed to 25 and 50 mg/mL, also affecting the egg viability. During the laying process, the eggs produced by the females exposed to the EO showed several morphological alterations such as dehydrated, darkened, and disaggregated, and these alterations were more severe as the concentrations increased. The mortality percentages were 58.9%, 70.8% and 92.7% when the ticks were exposed to 12.5, 25 and 50 mg/mL of E. viscosa oil, respectively. In the same concentrations, the efficacy of L. schaueriana was 39.3%, 53.4%, and 84.6%. Therefore, it can be concluded that the essential oils of E. viscosa and L. schaueriana have acaricidal effect in females of R. sanguineus s.l ticks.

Rhamnolipids and essential oils in the control of mosquito-borne tropical diseases.

The diseases transmitted by mosquito vectors are a great public health issue. Thus, effective vector control becomes the main strategy to reduce their prevalence. However, insecticide resistance has become a huge concern for the mitigation of mosquitoes; here, we propose the use of rhamnolipids in emulsion with clove oil against Aedes aegypti and Culex quinquefasciatus. The toxicity of rhamnolipids and clove oil to two species of mosquitoes transmitting tropical diseases was investigated. After 24 h, the LC50 was 140 mg/L when rhamnolipids were used and 154 mg/L when clove oil was used against Aedes aegypti larvae. In the case of Culex quinquefasciatus, the LC50 was 130 mg/L for rhamnolipids and 19 mg/L for clove oil. When the concentrations of the upper limits of one of the solutions (rhamnolipid or clove oil) were mixed, 100% mortality was obtained after 24 h. The bioassay of insecticidal action for solutions of rhamnolipids and clove oil in the lower limit, upper limit, and lethal concentration 50 to determine the effect on 50% of the population (KD50) achieved low results from KD50 to the upper limit compared to the other concentrations for both Aedes aegypti and Culex quinquefasciatus. The rhamnolipids and clove oil at the upper limit concentration had the greatest repellent activity against the two mosquito species. Bioassays using different concentrations of rhamnolipids revealed variations in the morphology of the intestinal epithelium (800 mg/L). A concentration of 900 mg/L led to the most severe morphological changes in the organization of the epithelium and the cells lining the intestines of these larvae. When larvae were exposed to a concentration of 1000 mg/L, the marginalization of chromatin in the nucleus of epithelial cells was very severe, indicating the onset of cell death.Key points• The toxicity of rhamnolipids and clove oil has a larvicidal, insecticidal, and repellent effect.• The combination of concentrations of these compounds enhances their action.• Different concentrations of rhamnolipids led to severe morphological changes in the organization of the epithelium and the cells and the intestines of larvae.

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.

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

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

Evaluation of Tagetes patula (Asteraceae) as an ecological alternative in the search for natural control of the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

The cattle tick Rhipicephalus microplus is a great sanitary problem and causes huge losses to livestock, being a vector of important diseases. The aim of this work was to verify the action of plant-derived material obtained from Tagetes patula on the life cycle of R. microplus, as well as to analyze the action of these extracts on ovary cells of engorged females. In the Adult Immersion Test, the crude ethanolic extract (TpEtOH) showed an efficacy of 99.2% (LC50 = 18.60 mg mL-1). From the repellency test, it was found that both the TpEtOH and essential oil were 100% efficient on the larval stage. In addition, in the Larval Package Test, a mortality of 98.37% was obtained with TpEtOH (LC50 = 3.798 mg mL-1). From the microscopic analysis of ovary cells, morphological changes in the chorion and cytoplasm of oocytes were observed, with vacuolization around the germinal vesicle and disorganization of the pedicular cells, suggesting an interference in the normal embryogenic formation of the larvae. These results demonstrate that T. patula extracts interfere at all stages of development of the ixodid, from eggs to adults forms, and have a pronounced repellent effect. In addition, the results of the cytotoxicity assays performed on keratinocytes, as well as previous information on oral and dermal acute toxicity (LD50 > 4000 mg kg-1), attest that T. patula can be a safe alternative for ectoparasitic control, representing an alternative for development of a formulation used to help control populations of ticks in the field.

Comparative analysis of germ cells and DNA of the genus Amblyomma: adding new data on Amblyomma maculatum and Amblyomma ovale species (Acari: Ixodidae).

Among tick species, members of the subfamily Amblyomminae have received special attention, since they serve as vectors for pathogens such as Rickettsia spp. and display cryptic species complexes that make their taxonomical classification challenging. Amblyomma ovale, Amblyomma maculatum, and other species of the genus Amblyomma have shown a long history of taxonomic controversies. Spermiotaxonomy has proved to be a valuable tool in the solution of systematic conflicts in Metazoa that can aid molecular and external morphological analyses in ticks and, overall, provide more robust analyses and results. With this in mind, this study included histological analyses of the reproductive system of the species A. ovale and A. maculatum, as well as the description of morphohistological characters of the male reproductive system of ticks of the genus Amblyomma, in order to evaluate these characters within the current clustering proposals. In addition, 16S rDNA and COI (mitochondrial) molecular markers were used to study the genetic relationships of the species. The results show that the tick male reproductive system and its germ cells contain useful candidate characters for taxonomical analyses of Ixodida.

Detrimental effect of deltamethrin on the central nervous system (synganglion) of Rhipicephalus sanguineus ticks.

Ticks are ectoparasites of medical and veterinary importance, which transmit many infectious agents, causing significant damage to the hosts. The "dog tick" Rhipicephalus sanguineus is responsible for transmitting several pathogens to dogs, motivating researchers to investigate efficient and sustainable control methods. Currently, chemical acaricides currently in use target the central nervous system (synganglion), which is responsible for controlling all the systemic functions of the ticks. Here, the neurotoxic potential of deltamethrin on the synganglion of unfed R. sanguineus female ticks was investigated. The results showed that the synganglion of the females belonging to the control group presented intact morphological characteristics; however, the ones from the treatment group (exposed to 1.5, 3.12 and 6.25 ppm of deltamethrin) displayed alterations, which were increasingly intense as the concentration increased. Observed alterations were mainly in the cortex region and in the neuropile, indicating that the deltamethrin is neurotoxic.

Deltamethrin as inductor agent of precocious ovarian degeneration in Rhipicephalus sanguineus s.l. (Acari: Ixodidae) ticks.

The cosmopolitan species Rhipicephalus sanguineus s.l. is one of the most widely distributed ticks all over the world. These ectoparasites are vectors of several pathogens and cause significant direct damage to their hosts. The biological success of these ectoparasites has been attributed to their ovaries and salivary glands, organs that ensure their survival in various environmental conditions. The importance of the ovaries in ticks is that, after mating, the individuals are able to lay approximately three thousand eggs. The present study had the objective to demonstrate the effects of deltamethrin obtained from the product Butox P CE 25® (MSD Saúde Animal, São Paulo, Brazil) on the ovarian development of R. sanguineus s.l. females. The chemical was tested in the concentrations of 25, 50, 100 and 200 ppm (respectively 80, 40, 20 and 10 times lower than the recommended by the manufacturer). Through the application of histological techniques and HE staining, the results showed that the deltamethrin was potentially able to modify the morphophysiology of the oocytes in all developmental stages, interfering in the vitellogenesis, causing intense vacuolation, cytoplasmic disorganization, and alterations in the chorion secretion. In addition, the chemical affected the germ vesicle of some oocytes, causing damages and hypertrophy, fragmenting the chromatin and forming bodies strongly stained by hematoxylin. Therefore, this study confirmed that the deltamethrin had an important action on the reproductive system of the R. sanguineus s.l. females, causing the precocious structural disorganization of the germ cells, consequently preventing the generation of new individuals.

Comparative analysis of spermatids of Rhipicephalus sanguineus sensu lato (Ixodidae) and Ornithodoros rostratus ticks (Argasidae): morphophysiology aimed at systematics.

The phylogenetic relationships among tick species (Acari: Ixodida) have been revisited by several researchers over the last decades. Two subfamilies, Rhipicephalinae (Ixodidae) and Ornithodorinae (Argasidae), deserve special attention. The male reproductive system morphology, as well as the ultrastructure of the germ cells, may provide important information for phylogeny and systematics of metazoan groups, with spermatozoa exhibiting characters that can be used for this purpose. With that information in mind, this study aimed at evaluating, through a comparative analysis, the morphology of the male reproductive systems and germ cells of ticks species Rhipicephalus sanguineus and Ornithodoros rostratus. In order to do that, histology and scanning electron microscopy techniques were used. The results have shown that despite the similarities in the general morphology of the male reproductive system among studied Ixodida so far, there are morphological differences among the species studied herein, mainly the U-shaped testis (ancestral character) in O. rostratus and the pair testes (derived character) in R. sanguineus, and the general morphology of germ cells (spermatids V). Besides that, the morphological changes observed during the spermiogenesis appear to be different between the species studied here, probably characterizing the two families considered. The data generated in this study showed the importance of comparative internal morphology studies, mainly in regard to spermatology, despite the morphological data obtained herein not being enough to product a cladogram (sperm cladistics), it was already possible to observe clear differences among families Argasidae and Ixodidae in regard to the organization of their male reproductive systems and concerning the external morphology of spermatids. Data yet to be obtained through transmission electron microscopy techniques will allow the application of spermiocladistics and spermiotaxonomy as tools for tick systematics.

Potential of the chemical dinotefuran in the control of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged female ticks.

Ticks are vectors of several pathogens to vertebrates, including the human being. They produce lesions on the hosts during the blood feeding and great economic losses. Several chemical acaricides have been used in an attempt to control tick infestations; however these substances are harmful to both the human being and non-target organisms, and to the environment. Therefore, there is a need to fight these ectoparasites through less harmful methods, less aggressive to the environment, non-target organisms and to the human health. The present study examined the efficacy of dinotefuran on the susceptibility of Rhipicephalus sanguineus semi-engorged females exposed to different concentrations of the product. Its lethal concentration of 50% (LC50) at 95% confidence interval was determined. The ticks were immersed in Petri dishes containing different concentrations of dinotefuran or distilled water for 5 minutes and then dried and maintained in an incubator for 7 days. The results showed the daily number of dead R. sanguineus semi-engorged females after being treated with different concentrations of dinotefuran. The mortality data in bioassay 2 were subjected to Probit analysis, where a LC50 of 10,182.253 ppm (8725.987-13,440.084) and 95% confidence interval were estimated. The susceptibility of R. sanguineus semi-engorged females to dinotefuran in higher concentrations of the acaricide was demonstrated, indicating that its effect is probably dose-dependent. In addition, the action of dinotefuran was slow and gradual, interfering in the development and growth of the individuals throughout the observation period (7 days).

Dynamics of cell and tissue genesis in the male reproductive system of ticks (Acari: Ixodidae) Amblyomma cajennense [corrected] (Fabricius, 1787) and Amblyomma aureolatum (Pallas, 1772): a comparative analysis.

Ticks are classified into three families: Argasidae, Ixodidae, and Nutalliellidae. The taxonomy and phylogeny within Ixodidae are still discussed by the specialists, thus requiring further studies. Amblyomma cajennese and Amblyomma aureolatum (Brazil) belong to two species complexes known as "cajennese" and "ovale", respectively, and are directly related to the transmission of the Brazilian spotted fever. This confirms the medical and veterinary significance of these species, as well as the need for further morphological studies that will bring a better understanding of their taxonomy, phylogeny, and control. In this context, the present study aimed to characterize the morphology of the male reproductive system of A. cajennese and A. aureolatum when unfed and after 4 days of feeding, thereby seeking to: (a) distinguish the two species or "complexes", and (b) study an internal system which has the potential to be targeted by acaricides. Therefore, males from both species (unfed and after 4 days of feeding) were cold-anesthetized, dissected, and had their reproductive systems removed for histological analysis. The results showed that the morphology of the male reproductive system is generally similar between both species, like in other Ixodidae ticks, exhibiting a multilobed accessory gland complex related to seminal fluid secretion, a pair of vasa deferentia and a pair of testes housing germ cells (spermatocytes) in different stages. The main differences were found in the development of the accessory gland complex cells and germ cells, showing that the maturation of the male reproductive system starts later in A. aureolatum, when compared to A. cajennese. However, during the blood meal, A. aureolatum development is increased, thus making germ cell maturation and gland complex activity higher than in A. cajennese. This study shows the differences in the development of the male reproductive systems of both species, while providing information that can assist in the establishment of new control methods.

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.

Histopathology of the tegument of rabbits infested by Rhipicephalus sanguineus (ACARI: IXODIDAE) ticks and exposed to selamectin (active principle of acaricide Revolution, Pfizer).

Ticks are hematophagous ectoparasites which can transmit several diseases to the host during their feeding process. When ticks mechanically damage the tissue, they eventually induce inflammatory responses on the skin spot where they are fixed. One of the alternatives to control these ectoparasites is the use of chemical substances like selamectin-the active principle of Pfizer's antiparasitic Revolution-a macrocyclic lactone capable of doing neurotoxic damage to the tick and eventually eliminating infestation in dogs and cats. The purpose of this study was to analyze, using histological and histochemical techniques, the occurrence of morphophysiological alterations in the skin of the host rabbits exposed to selamectin and infested with Rhipicephalus sanguineus (Acari: Ixodidae). Histologically, the exposed and infested rabbits showed a partial and/or total decrease in the stratum corneum and the epithelium decreased in the number of cell layers, consequently reducing the stratification (thinning) and quite pronounced formations of sub-epidermal edemas with consequent disorganization of collagen fibers in the dermal layer's connective tissue. Histochemical tests showed strong periodic acid-Schiff-positive reaction in the hair follicle and some regions of the dermis, besides resynthesis of collagen fibers detected by Mallory's trichrome technique. The obtained results showed that selamectin acts like a toxicant agent when in contact with the skin of the rabbit infested with ticks, inducing morphophysiological alterations in the acute inflammatory process in the animal's tegument. Selamectin is a chemical substance which has a dose-dependent action since higher concentrations cause greater morphophysiological damage in the skin of rabbits.

Inoculation of salivary gland extracts obtained from female of Rhipicephalus sanguineus (Latreille, 1806) (Acari, Ixodidae) with 2, 4, and 6 days of feeding in rabbit: I--histopathology of the feeding lesion.

This study analyzed the histopathology of rabbit skin, previously immunized with SGE2, SGE4, and SGE6 gland extracts prepared from salivary glands of Rhipicephalus sanguineus female with 2, 4, and 6 days of feeding, at the region of the R. sanguineus female feeding lesion 2, 4, and 6 days after tick attachment. In this work, infestation-naïve New Zealand White rabbits were inoculated either with the extracts (test group (TG)) or with phosphate buffer and complete Freund's adjuvant mixture (control group 2 (CG2)). Each extract-inoculated- (TG and CG2) and non-inoculated (CG1) rabbit was subsequently infested with R. sanguineus. Skin biopsies were collected from the rabbit at the tick feeding lesion at 2, 4, and 6 days of feeding. Results revealed that rabbit immunization with gland extracts induced acquisition of resistance against this species. It should be stated that the SGE4 extract was the most effective in developing an immune-inflammatory response against ectoparasites, being this process characterized by the presence of an early and intense inflammatory cell infiltrate. On the other hand, SGE6 extract caused a later appearance of resistance with less infiltrate occurrence and intense edema at the feeding lesion site. As to the inflammatory process deriving from SGE2 extract inoculation, it was the less intense. It was concluded that immunization with different extracts from R. sanguineus female salivary glands did not change microscope features of the inflammatory process, although an earlier or more intense and later response, which was also dependent on the inoculate extract, was noticed.

Fluazuron-induced morphophysiological changes in the cuticle formation and midgut of Rhipicephalus sanguineus Latreille, 1806 (Acari: Ixodidae) nymphs.

The present study demonstrated the effects of the arthropod growth regulator, fluazuron (Acatak®), in the formation of the integument and digestive processes of Rhipicephalus sanguineus nymphs fed on rabbits treated with different doses of this chemical acaricide. For this, three different doses of fluazuron (20, 40, or 80 mg/kg) were applied "pour on" to the hosts (groups II, III, and IV), as well as distilled water to the control group. On the first day after treatment (24 h), the hosts were artificially infested with R. sanguineus nymphs. After full engorgement (7 days), the nymphs were removed, placed on labeled Petri dishes, and kept in biochemical oxygen demand incubator for 7 days. The engorged nymphs were then taken for morphological, histochemical, and histological analyses. The results showed the occurrence of cytological, morphohistological, and histochemical alterations in the integument and midgut of nymphs from all the different treated groups. These alterations occurred at cuticular level in the subdivisions of the cuticle, related to the size of the digestive cells, amount of accumulated blood elements, and digestive residues, as well as the presence of vacuoles in the cytoplasm of the digestive cells. Thus, this study demonstrated that fluazuron acts on the integument and midgut cells of R. sanguineus nymphs fed on treated rabbits and pointed out the possibility of the use of this chemical-which is more specific, less toxic, and less harmful to the environment and nontarget organisms-in the control of R. sanguineus, at least in the nymphal stage of its biological cycle.

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.

Ecdysteroid levels changed by permethrin action in female Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) ticks.

As recent studies have shown that ecdysteroids may play a major role in the regulation of vitellogenesis in Ixodidae, the present study quantified, by means of a radioimmunoassay, the levels of ecdysteroids present in the hemolymph of semi-engorged females of Rhipicephalus sanguineus ticks obtained from control females (exposed to distilled water) and those exposed to increasing concentrations of permethrin. The levels of ecdysteroids decreased significantly as the concentration of permethrin increased, suggesting that this compound could be an inhibitor of ecdysteroids secretion, and consequently interfering with the reproductive ability of these ticks, since this hormone is responsible for the synthesis and incorporation of vitellogenin by oocytes. This study complements the previous results with R. sanguineus semi-engorged females, showing that permethrin is a potent agent causing major morphological changes in tick oocytes, such as the appearance of large vacuoles in the cytoplasm, reduction in the amount of yolk granules and a decrease in oocyte size, thus culminating in cell death and consequently reducing or preventing reproduction in treated females. The findings that permethrin leads to a decrease in ecdysteroid titers could represent an entry step into this scenario.