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This retrospective study aimed to evaluate potential hematological alterations in dogs treated with four different antineoplastic protocols. We selected 69 dogs that received at least four chemotherapy sessions and divided them into four groups: vinblastine (VBL) (n = 30), doxorubicin (DOX) (n = 15), combined CHOP protocol (cyclophosphamide, doxorubicin, vincristine, and prednisone) (n = 11), and vincristine group (VCR) (n = 13). Hematological parameters were recorded from blood samples collected in five periods: T0 (before the start of treatment), T1, T2, T3, and T4 (before each session). The VBL group showed a significant reduction in erythrocyte (p

Made available in DSpace on 2018-12-11T16:53:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-07-04 The present study evaluated the efficacy of Acmella oleracea extract, and the susceptibility of semi-engorged R. sanguineus females exposed to different concentrations of the extract, with lethal concentration 50 % (LC50) and confidence interval 95%. The ticks were mounted on Petri dishes and immersed in the different concentrations of the extract, distilled water or ethanol 50% + DMSO 1% for 5 minutes, dried and kept in BOD incubator for 7 days. The results showed the daily mortality rate among semi-engorged R.sanguineus females at different concentrations of the extract. The mortality data obtained in bioassay 2 were subjected to Probit analysis, through which a LC50 of 24,883 mg/ml (limits: 22,331 to 28,003 mg/ml) was determined, at a 95% confidence interval. Thus, this study demonstrated the sensitivity and susceptibility of semi-engorged R.sanguineus females to Acmella oleracea extract. The acaricidal potential was effective as from the application of the extract at the concentration of 12.5 mg/ml and was dose-dependent. i.e., higher mortality rates were observed as the concentration increased, with LC50 of 24,883 mg/ml reaching 100% at the concentration of 100 mg/ml in 24h. Department of Biology Institute of Biosciences São Paulo State University-UNESP Division of Chemistry of Natural Products CPQBA/UNICAMP Graduate Program in Animal Science School of Agricultural Science and Veterinary Medicine The Pontificia Universidade Catolica do Parana–PUCPR Embrapa Meio-Norte Department of Biology Institute of Biosciences São Paulo State University-UNESP

The tick vector Rhipicephalus sanguineus is established as a complex of closely related species with high veterinary-medical significance, in which the presence of different genetic, morphological, and biological traits has resulted in the recognition of different lineages within taxa. One of the most striking differences in the “temperate” and “tropical” lineages of R. sanguineus (s.l.) is the vector competence to Ehrlichia canis, suggesting that these ticks tolerate and react differently to pathogen infection. The present study addresses the SG and MG proteome of the R. sanguineus tropical and temperate lineages and compares their proteomic profile during E. canis infection. Batches of nymphs from the two lineages were allowed to feed on naïve and experimentally E. canis infected dogs and after molting, adults were dissected, and salivary glands and midgut tissues separated. Samples were screened for the presence of E. canis before proteomic analyses. The representation of the proteins identified in infected and non-infected tissues of each lineage was compared and gene ontology used for protein classification. Results highlight important differences in those proteomic profiles that added to previous reported genetic, biological, behavioral, and morphological differences, strengthening the hypothesis of the existence of two different species. Comparing infected and non-infected tissues, the results show that, while in midgut tissues the response to E. canis infection is similar in the salivary glands, the two lineages show a different pattern of protein representation. Focusing on the proteins found only in the infected condition, the data suggests that the cement cone produced during tick feeding may be implicated in pathogen infection. This study adds useful information to the debate on the controversial R. sanguineus systematic status, to the discussion related with the different vectorial competence occurring between the two lineages and identifies potential targets for efficient tick and tick-borne disease control., This research was supported by the project PTDC/CVT-WEL/1807/2014, subsidized by the Fundação para a Ciência e Tecnologia (FCT). The authors would like to acknowledge FCT for funds to GHTM – UID/Multi/04413/2013, Fundação de Amparo a Pesquisa do Estado de São Paulo - FAPESP (2018/06651-7), Conselho Nacional de Desenvolvimento Cientı́fico e Tecnológico –CNPq (421980/2016-8) and Coordenação de Aperfeiçoamento de Pessoal de Nı́vel Superior – CAPES (88887.464801/2019-00). MA is a fellowship of CNPq (Productivity Grant - Process #302420/2017-7). JF and JC are fellowships of FCT (Grants SFRH/BD/122894/2016 and SFRH/BD/121946/2016).

Ticks and tick-borne diseases are of global importance and can cause serious economic losses to cattle-raising businesses. However, only few attempts have been made to describe the relationships between various cellular immune components and tick counts of, and therefore resistance to, Rhipicephalus microplus. The present study evaluated the late inflammatory cellular response and dermal dendritic cells at the attachment site in naturally presensitized bovines, artificially infested by R. microplus ticks. Twenty-two crossbred (Holstein × Gir) male experimental cattle were artificially infested with approximately 10,000 larvae of R. microplus. Parasitic infestation was determined by counting the engorged female ticks (4.5-8.0 mm in diameter) at the 21st day after artificial infestation. Biopsies (6 mm) were taken prior to the infestation and along with the tick count from the right pinna of each animal, and examined with histological techniques and immunohistochemistry using S100 protein. Inflammatory cell counts were conducted in the sections stained with the May-Grünwald Giemsa technique, and immunostained dermal dendritic cells were evaluated and classified in scores ranging from 0 to 4. An average of 39.2 ticks per animal was found. Eosinophils (47.8%) constituted the major portion of the cellular infiltrate, followed by mononuclear cells (28.3%), neutrophils (14.4%), and basophils (0% to approximately 2%). The dendritic cell count shows a considerable population in the dermis, with pre- and post-infestation mean scores of 1.54 and 1.89, respectively; these scores were not significantly different. Our results pointed out the importance of the cellular response in the cattle resistance to ticks.

The present experiment was conducted to evaluate and compare the impact of Ageratum conyzoides plant extract (ACE) with routinely used synthetic acaricides i.e., amitraz and coumaphos on the oogenesis of engorged adult females of Rhipicephalus microplus tick. On the day of dropping from the host, panoistic ovary of R. microplus appeared white in colour, horseshoe shaped, hollow tubular organ with immature oocytes predominantly in dorsal groove. Different developmental stages of oocytes (I–V) proceed simultaneously and asynchronously. Oocytes showed gradual increase in size, deep brown colored with accumulation of eggs in oviduct during 24–72 hours of development.At LC90 concentration a highly significant (p

A synthetic 20 amino acid peptide of the ribosomal protein P0 from ticks, when conjugated to keyhole limpet hemocyanin from Megathura crenulata and used as an immunogen against Rhipicephalus microplus and Rhipicephalus sanguineus s.l. species, has shown efficacies of around 90%. There is also experimental evidence of a high efficacy of this conjugate against Amblyomma mixtum and Ixodes ricinus species, which suggest that this antigen could be a good broad-spectrum anti-tick vaccine candidate. In this study, the P0 peptide (pP0) was chemically conjugated to Bm86 as a carrier protein. SDS-PAGE analysis of this conjugate demonstrated that it is highly heterogeneous in size, carrying from 1 to 18 molecules of pP0 per molecule of Bm86. Forty-nine out of the 54 lysine residues and the N-terminal end of Bm86 were found partially linked to pP0 by using LC-MS/MS analysis and the combination of four different softwares. Several post-translational modifications of Bm86 protein were also identified by mass spectrometry. High immunogenicity and efficacy were achieved when dogs and cattle were vaccinated with the pP0&ndash, Bm86 conjugate and challenged with R. sanguineus s.l. and R. microplus, respectively. These results encourage the development of this antigen with promising possibilities as an anti-tick vaccine.

Infrared thermography has been used to help in diagnosing lameness. It is hypothesized that, if used in a routine basis, it could help in understanding musculoskeletal modifications during race training. This study aimed to evaluate thermal variation in the musculoskeletal regions of young Thoroughbred (TB) horses during their initial months of race training. Thermographic examinations were performed once every 2 weeks on 16 (10 male, 6 female) two-year-old TB racehorses, from arrival to the racetrack in June 2016, until January 2017, for a total of 16 evaluations. Thermographic imaging was performed using the appropriate protocol. Temperature (°C) was measured at the dorsal and palmar/plantar aspects of specific regions of interest (fetlock, metacarpal, metatarsal, carpal, tarsal, thoracolumbar, sacroiliac spine, and both hips). Initially, we found a thermal balance and all regions demonstrated a positive correlation with one another. However, a significant difference was noted between the left and right sides as training progressed. Four horses were withdrawn from the study after 50% of evaluations because of metacarpal conditions associated with training. Thermographic examination revealed changes before the clinical manifestation of these conditions. In conclusion, this study demonstrated that infrared thermography is an image technique that can facilitate understanding of musculoskeletal system modifications to race training and should be further investigated as a predictive tool to anticipate the occurrence of lesions.

Leprosy is an infectious disease caused by Mycobacterium leprae and frequently resulting in irreversible deformities and disabilities. Ticks play an important role in infectious disease transmission due to their low host specificity, worldwide distribution, and the biological ability to support transovarial transmission of a wide spectrum of pathogens, including viruses, bacteria and protozoa. To investigate a possible role for ticks as vectors of leprosy, we assessed transovarial transmission of M. leprae in artificially-fed adult female Amblyomma sculptum ticks, and infection and growth of M. leprae in tick cell lines. Our results revealed M. leprae RNA and antigens persisting in the midgut and present in the ovaries of adult female A. sculptum at least 2 days after oral infection, and present in their progeny (eggs and larvae), which demonstrates the occurrence of transovarial transmission of this pathogen. Infected tick larvae were able to inoculate viable bacilli during blood-feeding on a rabbit. Moreover, following inoculation with M. leprae, the Ixodes scapularis embryo-derived tick cell line IDE8 supported a detectable increase in the number of bacilli for at least 20 days, presenting a doubling time of approximately 12 days. As far as we know, this is the first in vitro cellular system able to promote growth of M. leprae. Finally, we successfully transformed a clinical M. leprae isolate by inserting the reporter plasmid pCHERRY3; transformed bacteria infected and grew in IDE8 cells over a 2-month period. Taken together, our data not only support the hypothesis that ticks may have the potential to act as a reservoir and/or vector of leprosy, but also suggest the feasibility of technological development of tick cell lines as a tool for large-scale production of M. leprae bacteria, as well as describing for the first time a method for their transformation., Author summary Leprosy is a slow-progressing and extremely debilitating disease; the armadillo is the only animal model able to mimic the symptoms observed in humans. In addition, the causative agent, Mycobacterium leprae, is not cultivable in vitro. Due to these constraints the chain of transmission is still not yet completely understood. We know, however, that at least two animals, armadillos in the Americas and red squirrels in the UK, are natural reservoirs of the bacillus, although their role in disease epidemiology is unclear. This information raised the following question: Can ticks carry leprosy from wild animals to humans? In the present study we demonstrated that artificially-infected female cayenne ticks are able to transmit the bacillus to their offspring, which were then able to transmit it to rabbits during bloodfeeding. We were able to grow M. leprae in vitro in a tick cell line for the first time. We also generated the first transgenic M. leprae strain, making the pathogen fluorescent in order to monitor its viability in real time. We believe that this new methodology will boost the screening of new drugs useful for control of leprosy, as well as improving understanding of how M. leprae causes disease.

INCOGARR is a thematic network recently approved to be financially supported by the Ibero-American Program of Science and Technology for Development (CYTED). The objectives of this Network are the design and evaluation of an efficient and feasible anti-tick vaccine candidate from the technical and economical points of view and also sharing experiences in the immunological control of ticks as part of an Integrated Control Program. The Network consists of seven laboratories and one company from six countries. The first meeting of the Network took place with the representation of each laboratory involved. In the meeting, general and specific objectives and activities of the Network were discussed and it was a very nice example of international collaboration to address an unsolved worldwide topic on tick control in which laboratories with different competencies and expertise join their efforts in a common goal.

Comparative studies between brown dog tick Rhipicephalus sanguineus populations from Brazil (Jaboticabal, São Paulo) and Argentina (Rafaela, Santa Fé) showed significant biological, morphological and genetic differences between them. This work aimed to study, in a comparative way, the acquisition of resistance in domestic dogs to R. sanguineus from Jaboticabal and Rafaela, after successive and controlled infestations. Ticks were kept in a BOD incubator under controlled conditions (27 °C, 80 % relative humidity, 12-h photoperiod). Ten dogs, Dachshund breed, males and females, 6 months old, short- or long-haired, without prior contact with ticks, were used as hosts. They were distributed into two experimental groups composed of five animals each: G1 infested with ten adult couples of R. sanguineus (Jaboticabal) per animal, and G2 infested with ten adult couples of R. sanguineus (Rafaela) per animal. Ticks' biological parameters and titration of antibodies from the dogs' sera by ELISA test were used for comparison between the strains. Results of the biological parameters showed that the dogs did not acquire immunity to either of the R. sanguineus strains after repeated infestations. The ELISA test showed low antibody titers in sera of dogs from G2, in successive infestations, and higher antibody responses post second and third infestations in G1. It also demonstrated cross-reactivity between sera of dogs infested with R. sanguineus (Jaboticabal) and antigens from R. sanguineus (Rafaela) and vice versa. We conclude that Dachshund dogs did not develop resistance against neither Jaboticabal nor Rafaela strains of R. sanguineus.

This study presents the morphological and physiological characterization of the salivary glands of semi-engorged Amblyomma triste females. Unfed individuals were placed on New Zealand White rabbits for feeding and the females, after 4 days, were collected, dissected and the salivary glands were submitted to the application of histological (hematoxylin–eosin technique) and histochemical tests for the detection of protein (bromophenol blue technique, polysaccharides (periodic acid–Schiff technique), lipid (Nile blue technique) and calcium (von Kossa technique). The histological results show that the glandular tissue is composed by a system of ducts and three types of acini (I, II and III). The acini I are formed by a large central cell surrounded by several smaller agranular peripheral cells. Acini II are formed by cells a , c1 , c2 , c3 and c5 , which are full of secretion granules. Acini III are constituted by cells d , e and f ; the former two contain secretion granules, the latter is agranular. The glandular histochemical composition was also verified. Data obtained here will certainly help in the understanding of the cellular morphology and of the general physiology of these organs in this specie, providing important information for the creation of scientific bases which will contribute for the development of more specific and efficient methods of control.

This study evaluated a recombinant aquaporin 1 protein of Rhipicephalus (Boophilus) microplus (RmAQP1) as antigen in a vaccine against R. sanguineus. Five dogs were immunized with RmAQP1 (10 µg) + adjuvant (Montanide) (G1), and five were inoculated with adjuvant only (G2), three times. Twenty-one days after the last immunization, animals of both groups were challenged with R. sanguineus larvae, nymphs and adults, and their biotic potential was compared. Blood samples were collected before each immunization and every 28 days after the last immunization for 10 weeks. Serum antibody titers (IgG) were assessed by ELISA. We observed that: engorgement period of adult females from G1 was 12% shorter than G2; larvae from G1 had 8.7% longer engorgement period than G2 and weighed 7.2% less; nymphs from G1 had 4.5% shorter engorgement period than G2 and weighed 3.6% less; although the antibody titers increased following the second immunization, they rapidly decreased after the third immunization. Results indicated low immunoprotection of RmAQP1 against adult R. sanguineus ticks, and possible efficacy on larvae and nymphs fed on immunized dogs. Further studies should be performed for a full evaluation of the immunoprotection of RmAQP1 against R. sanguineus infestations in dogs. Resumo Este estudo avaliou a proteína recombinante (aquaporina) do carrapato Rhipicephalus (Boophilus) microplus como antígeno em vacina contra Rhipicephalus sanguineus. Cinco cães foram imunizados com RmAQP1 (10 µg) + adjuvante (G1) e cinco foram inoculados apenas com adjuvante (G2), três vezes. 21 dias após a última imunização todos os animais foram desafiados com larvas, ninfas e adultos de R. sanguineus, e potencial biótico dos carrapatos foi comparado. Amostras de sangue foram coletadas antes de cada imunização e a cada 28 dias após a última imunização, durante 10 semanas. Títulos de anticorpos dos soros dos cães foram avaliados por ELISA. Resultados: o período de ingurgitamento das fêmeas do G1 foi 12% mais curto que o período de ingurgitamento de G2; o período de ingurgitamento das larvas do G1 8,7% foi mais longo e o peso 7,2% menor que no caso de G2; o período de ingurgitamento das ninfas do G1 4,5% foi mais curto e peso 3,6% menor que no caso do G2; aumento dos títulos de anticorpos do G1 após a segunda imunização e declínio após a terceira imunização. Os resultados indicaram baixo potencial de imunoproteção de RmAQP1 contra R. sanguineus adultos, e possível eficácia contra larvas e ninfas, na dose testada. Sugere-se desenvolver novos estudos para melhor avaliação da eficácia de RmAQP1 contra R. sanguineus em cães.

This is an ultrastructural study of two important organs of ticks: the integument, covering the body and involved in the molting process, and the midgut, the first organ to have contact with the ingested host blood. The integument of Rhipicephalus sanguineus sensu lato freshly engorged nymphs consists of a layer of epidermal cells rich in organelles and a cuticle, subdivided in epicuticle and procuticle. The latter presents two distinct regions: the exocuticle (with several pore canals) and the endocuticle (secreted in highly organized overlaid lamellae). The midgut of R. sanguineus sensu lato freshly engorged nymphs consists of an epithelial wall formed by two types of cells: the generative cells, with few organelles; and the digestive cells, with several endosomes, digestive vacuoles, hematin residual bodies, and small drops of lipid.

The sloth's giant tick Amblyomma varium Koch, which is a neotropical species that inhabits tropical rainforests, is the largest tick reported to date. The adult stage of this tick parasitizes mammals from the families Bradypodidae and Magalonychidae (Xenarthra) nearly exclusively. This study aimed to describe morphological and histological features of the reproductive system and the oocyte maturation process of this tick species. The ovary of A. varium is a long single tubular organ that is horseshoe-shaped, winding and arranged in the posterior part of the body. Two oviducts are connected to the ovary on each side; these thicken at certain region forming the uterus (common oviduct), followed by a muscular connecting tube, vagina and genital aperture. A large number of oocytes at different stages of development are attached to the ovary wall by the pedicel, as they reach maturity they are released into the ovary lumen and from there to the genital aperture. These oocytes develop simultaneously and asynchronically along the ovary. Amblyomma varium oocytes were classified into five development stages (i.e., I-V), and specific characteristics were observed; the processes of yolk and chorion deposition begin early in oocytes stage II, and oocytes V exhibit a very thick chorion and eggs of a large size. These characteristics are likely adaptations that enhance the survival and the reproductive success of this extremely host-specific tick, which is limited to a particular environment.

Rhipicephalus sanguineus is a species of tick which is widely distributed in America, Africa and Australia and is probably the most prevalent among all the other ixodid tick species. The present study demonstrated the effects of the arthropod growth regulator fluazuron (Acatak(®)), in the formation of the integument and the digestive processes of R. sanguineus nymphs fed on rabbits treated with different doses of this chemical acaricide. For this, three different doses of fluazuron (20mg/kg, 40mg/kg and 80mg/kg) were applied "pour on" to the hosts divided into three different treated-groups (II, III, IV) of three animals each. A fourth group (I) of rabbits (n=3) was given distilled water as control. On the first day after treatment (24h), the hosts were artificially infested with R. sanguineus nymphs. After full engorgement (7 days), the nymphs were removed and placed on labeled Petri dishes and kept in BOD incubator for 7 days. The engorged nymphs were then taken to ultra-structural analysis. Results revealed the following main ultra-structural changes in the nymphs integument and midgut of the different treated groups (II, III, IV): cuticle disorganization and the absence of subdivisions, damages in the integument epithelial cells, size of digestive cells, amount of endosomes, autophagic and digestive vacuoles, accumulated digestive residues, lipid droplets and organelles found in the digestive cells' cytoplasm, as well as the presence of microvilli in their plasma membranes. It is concluded that fluazuron may act on the integument and midgut cells of R. sanguineus engorged nymphs by impairing the synthesis of the new cuticle and the digestive processes (absorption of the blood ingested from the host, digestion - hemolysis, formation of digestive residues and release of nutrients to be converted into lipid, as well as for the synthesis of structural protein), which interfere in the development of nymphs, being able to prevent the emergence of adults after periodical ecdysis. These data indicate the possibility to use this arthropod growth regulator (AGR) in the control of R. sanguineus, at least in the nymphal stage of its biological cycle.

This study performs the ultrastructural description of the central nervous system (CNS) of Rhipicephalus sanguineus larvae and nymphs. The CNS is surrounded by the neural lamella, a thin acellular sheath. Just below, the perineurium is found, formed by glial cells, with irregular membranes, elongated nucleus and few organelles in the cytoplasm. Internally, the synganglion is divided into an outer cortex, containing the cellular bodies of the neural cells and an inner neuropile. The neural cells can be classified into two types. Type I is most frequently observed, presenting spherical or oval shape and a large nucleus occupying most part of the cytoplasm, in which few organelles are observed. Type II cells are polygonal, presenting a great cytoplasmic volume and the nucleus dislocated towards the cellular periphery. The subperineurium is located between the cortex and the neuropile. The neuropile is the innermost region of the synganglion, the most complex one and formed by extensions of the neural cells. ...

The present study analyzed, by means of confocal laser scanning microscopy, the epidermis of rabbits infested by the brown dog tick Rhipicephalus sanguineus and exposed to concentrations of 50%, 80% and 100% of a selamectin-based commercial acaricide (Pfizer's Revolution ® ). The results demonstrated that rabbits exposed to concentrations of 80% and 100% of the Revolution acaricide, which contains 12% selamectin, showed thinning of epithelial tissue of the epidermis with associated cellular disorganization. Individuals exposed to a 50% concentration showed lower epidermal tissue disorganization when compared to those exposed to the higher doses of the acaricide (80% and 100%). Whereas selamectin, when used in higher concentrations (80% and 100% Revolution ® ) can alter the morphology of the epidermis, at lower concentrations (50%), even though still able to eliminate ectoparasites, it causes less toxicity damage to the host. Selamectin can be considered a dose-dependent toxic agent, since higher concentrations increase the morphological changes in the epidermis of the host rabbits.

Because of the high incidence of thromboembolic diseases in humans, experimental models of thrombosis have been widely developed in different animal species. The pathogenesis of thrombosis is associated with three components, first outlined by Virchow in 1856: vessel injury, stasis, and hypercoagulability. Based on this concept, the purpose of the present investigation was to create an innovative model of jugular thrombophlebitis in horses that included components of Virchow's triad and excluded surgical procedures. Eighteen horses were subjected to blood vessel injury through the coadministration of sclerosing agents (glucose and ethanolamine oleate) and transitory occlusion of the jugular flow by manual compression. Thrombus formation was followed by ultrasonography imaging, and all horses developed jugular thrombophlebitis, showing that the proposed model was effective. Once occlusive thrombophlebitis was induced, jugular venous pressure cranial to the lesion was evaluated and yielded increased values, suggesting cephalic hypertension. Biochemical tests were performed to verify hepatotoxicity and nephrotoxicity after the ethanolamine injection, but no abnormalities were observed. Five horses were then euthanized to evaluate the vascular, hepatic, and renal tissues. The jugular vein wall had increased thickness, inflammatory cell influx, endothelial destruction, and thrombus firmly adhered to the vessel intima. Histological evaluation of the hepatic and renal tissues was normal. The present thrombophlebitis model in the jugular vein of the horse is simple and reproducible, providing a useful tool for investigating acute and chronic venous thrombosis because the model allows evaluation of different aspects of the prevention, pathogenesis, and treatment of this disease.

The present study investigated the effects of different concentrations of Acmella oleracea extract on the germinative cells and digestive processes of semi-engorged Rhipicephalus sanguineus females. For this experiment, 150 ticks were divided into five groups (30 individuals each). The animals were immersed for 5 min in different concentrations of the extract, distilled water, or ethanol 50%/DMSO 1%, dried, and kept in biological oxygen demand incubator for 7 days. The alterations were associated with the size of germinative cells and yolk granules; presence, size, and location of vacuoles in the cytoplasm of germinative cells; nuclear modifications in the germinative cells; damages to the nucleus and cytoplasm of the midgut generative cells; size of digestive cells; number of captured blood elements; accumulated digestive residues and digestive vacuoles in the digestive cells cytoplasm; and the number and distribution of proteins and polysaccharides in all the cells of both organs. The concentrations used in this study prevented an efficient and complete blood digestion by the midgut epithelial cells of the treated animals, resulting in the absence of the necessary nutrients to maintain the physiological events in the ectoparasites. In advanced stages, This can lead the ectoparasite to death. The germinative cells were highly impaired and probably not able to advance developmental stages (I–V) or complete vitellogenesis to be released during ovulation, which would prevent the females from originating a new individual. Thus, it can be concluded that the effects of A. oleracea are similar to those caused by chemical products widely recognized as effective to control ticks.

The present study evaluated the efficacy of fluazuron (active ingredient of the acaricide Acatak®) and its effects on Rhipicephalus sanguineus nymphs fed on rabbits exposed to different doses of this insect growth regulator. Three different doses of fluazuron (20 mg/kg, 40 mg/kg, and 80 mg/kg) were applied on the back of hosts (via "pour on"), while distilled water was applied to the Control group. On the first day of treatment with fluazuron (24 h), hosts were artificially infested with R. sanguineus nymphs. Once fully engorged, nymphs were removed and placed in identified Petri dishes in Biochemical Oxygen Demand (BOD) incubator for 7 days. After this period, engorged nymphs were processed for ultramorphological analysis. The results revealed alterations in the ultramorphology of many chitinous structures (smaller hypostome and chelicerae, less sclerotized scutum, fewer sensilla, fewer pores, absence of grooves, marginal and cervical strips and festoons in the body, even the anal plaque was damaged) that play essential roles for the survivor of ticks and that can compromise the total or partial development of nymphs and emergence of adults after periodic molting. Our findings confirm the efficacy of fluazuron, a more specific and less aggressive chemical to the environment and human health, and that does not induce resistance, in nymphs of the tick R. sanguineus in artificially infested rabbits treated with this arthropod growth regulator (AGR), indicating that it could be used in the control of this stage of the biological cycle of the tick R. sanguineus.

Actually, the most used method to control ticks is synthetic acaricides with neuro- toxic action. However, the use of these methods presents inconveniences, such as the contamina- tion of the environment and risks to the host's health due to the residual effects. Thus, several studies have been developed aiming to fi nd 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 pres- ent study aimed to analyze the action of andiroba oil (Carapa guianensis) on the synganglion of Rhipicephalus sanguineus semiengorged females through morphological and cytochemical tech- niques 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 perineu- rium and the cortical region, vacuolated cortex cells and with cell swelling, neural cells with pic- notic 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. Microsc. Res. Tech. 00:000-000, 2013. V C 2013 Wiley Periodicals, Inc.

GAVAC (Heber Biotec S.A, Havana, Cuba) is a commercially available vaccine developed with the Rhipicephalus (Boophilus) microplus Bm86 recombinant antigen. Bm86 is a “concealed” antigen that is present in the plasmatic membrane of tick gut epithelial cells with unknown function so far. It is well known that after vaccination in the last fifteen years in Cuba, there was a significant decrease of babesiosis (Babesia bovis and Babesia bigemina) and anaplasmosis (Anaplasma marginale) in cattle. A reduced transmission capacity of ticks fed on tick-immune animals and humans has been reported for several tick-borne pathogens. Recent experiments have demonstrated that an anti-tick vaccine may contribute to the control of tick-borne pathogens not only by decreasing the exposure of susceptible hosts to ticks, but also by reducing the vector capacity of ticks. In this study, the potential of Bm86 vaccination to interfere with pathogen transmission among ticks was evaluated by using as experimental model the brown dog tick Rhipicephalus sanguineus and the tickborne Babesia canis and Ehrlichia canis pathogens. Dogs, vaccinated and not vaccinated, were infested with pathogen-infected ticks and noninfected nymphs of R. sanguineus. After feeding, the pathogen transmission to newly molted adults from co-feeding uninfected nymphs was studied by conventional PCR and qPCR. Results suggest that the anti-Bm86 antibodies could be able to block the transmission of B. canis and/or E. canis from infected to non-infected ticks.

Made available in DSpace on 2018-12-11T17:26:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-02-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The present study demonstrated the effects of dinotefuran (active compound of the Protetor Pet® acaricide) in germ cells and the digestive processes of semi-engorged females of R. sanguineus exposed to different concentrations of the chemical. For this purpose, 120 semi-engorged females were divided into four treatment groups with 30 individuals each: group I or control (distilled water), group II (5000 ppm), group III (6250 ppm), and group IV (8334 ppm of dinotefuran). All ticks were immersed in different concentrations of dinotefuran or in distilled water for 5 min and then were dried and stored in biological oxygen demand (BOD) incubator for 7 days. The results show the action of this compound, exhibiting morphohistologic and histochemical changes in the oocytes and the midgut cells of individuals of different groups, which were compared with those of group I (control). The alterations occurred mainly in relation to the size of the germ cells and yolk granules; presence, quantity, size, and location of vacuoles found in the cytoplasm of these germ cells; the damage occurred in the generative cells of the midgut; the size of the digestive cells; the quantity of blood elements captured, accumulated digestive wastes and digestive vacuoles found in the cytoplasm of the digestive cells of the midgut, as well as the amount and distribution of proteins, polysaccharides, lipids of all cells in both organs. So, it has demonstrated the effectiveness of dinotefuran in the reduction of fertility and digestive processes of semi-engorged females of R. sanguineus, data that points the possibility of employing this chemical to control these ectoparasites. Department of Biology Institute of Biosciences São Paulo State University-UNESP, Av. 24 A, n° 1515, Postal Code 199 Department of Animal Pathology São Paulo State University-UNESP, Via de Acesso Prof. Paulo Castellane, s/n Graduate Program in Animal Science School of Agricultural Science and Veterinary Medicine The Pontificia Universidade Catolica do Parana—PUCPR, Rua Imaculada Conceição, 1155 Department of Biology Institute of Biosciences São Paulo State University-UNESP, Av. 24 A, n° 1515, Postal Code 199 Department of Animal Pathology São Paulo State University-UNESP, Via de Acesso Prof. Paulo Castellane, s/n

In this study, different geographical populations of Rhipicephalus sanguineus sensu lato were compared by molecular, biological, and morphometric methods. Phylogenetic trees were constructed using 12S and 16S rDNA sequences and showed two distinct clades: one composed of ticks from Brazil (Jaboticabal, SP), Cuba (Havana) Thailand (Bangkok) and the so-called "tropical strain" ticks. The second clade was composed of ticks from Spain (Zaragoza), Argentina (Rafaela, Santa Fe) and the so-called "temperate strain" ticks. Morphometric analysis showed good separation between females of the two clades and within the temperate clade. Males also exhibited separation between the two clades, but with some overlap. Multiple biological parameters revealed differences between the two clades, especially the weight of the engorged female. These results confirm the existence of at least two species under the name "R. sanguineus". Fil: Sanches, Gustavo S.. Universidade de Sao Paulo; Brasil Fil: Évora, Patrícia M.. Universidade de Sao Paulo; Brasil Fil: Mangold, Atilio Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina Fil: Jittapalapong, Sattaporn. Kasetsart University; Tailandia Fil: Rodriguez Mallon, Alina. Centro de Ingeniería Genética y Biotecnología-CIGB; Cuba Fil: Guzmán, Pedro E. E.. Centro de Ingeniería Genética y Biotecnología-CIGB; Cuba Fil: Bechara, Gervásio H.. Pontifícia Universidade Católica do Paraná; Brasil. Universidade de Sao Paulo; Brasil Fil: Camargo Mathias, Maria I.. Universidade de Sao Paulo; Brasil

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.

KEY WORDS ticks; natural product; germ cells; cytotoxicity; reproductive efficiency indexABSTRACT Because of the increasing medical-veterinary importance of ticks, the developmentof alternative control methods, less aggressive to the host and the environment has become the tar-get of several researches. In this sense, the present study analyzed the action of different concen-trations (5, 10, and 20%) of andiroba seed oil (Carapa guianensis) on the reproductive system ofRhipicephalus sanguineus females, through histochemical techniques and the quantification of thereproductive efficiency index. The results showed that andiroba oil is a potent natural agent, ableto cause several changes in the oocytes of this species, impairing the reproductive success, oncethis 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 compo-nents are essential for the viability of the embryo. In addition, it was observed that this productstimulate the oviposition, mainly at the concentration of 20%. This higher production of eggs repre-sents a defense mechanism developed by the organism in order to ensure the reproductive successof the species, even in the presence of the toxic agent. However, the results obtained suggested thatthe laid eggs would not be viable, due to the great changes undergone by the oocytes. Thus, thepresent study showed that the use of this vegetal product would be an alternative way to controlthe 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. Microsc. Res.Tech. 00:000–000, 2012.

Pyrethroids such as permethrin are synthetic compounds widely used in the agriculture of many countries to combat plagues and in domestic products, such as acaricides. Not so long ago these chemicals were characterized as non-toxic for non-target organisms; however, recent studies have showed that these compounds could present toxic potential for many organisms. In this sense, this study presents genotoxic and mutagenic potential of per- methrin administered intraperitoneally in mice under artificial conditions by the use of micro- nucleus assay in the peripheral blood of these animals. The mice were divided into five groups: group I 5 negative control (distilled water), group II 5 positive control (cyclophosphamide), group III 5 30% of permethrin LD50 (96 mg/kg), group IV 5 50% of permethrin LD50 (160 mg/ kg), and group V 5 80% of permethrin LD50 (256 mg/kg). The peripheral blood was collected 24, 48, and 72 h after treatment. Results showed that all the tested permethrin dosages pre- sented genotoxic and mutagenic effects 24 h after treatment, which would contradict the clas- sification of this chemical product as moderately toxic, i.e., unable to cause damages to the cell DNA. Microsc. Res. Tech. 00:000-000, 2012. V C 2012 Wiley Periodicals, Inc.

Ovarian development and egg maturation are essential stages in animal reproduction. For bisexual ixodid ticks, copulation is an important prerequisite for the completion of the gonotrophic cycle. In this study, we aimed to characterize the morpho-histological changes in the ovary and oocytes of the tick Rhipicephalus sanguineus, together with the identification of feeding and reproductive parameters associated with mating. Virgin and cross-mated females (with R. turanicus males) weighed 60% less at full engorgement than females mated conspecifically. In addition, the oocytes of these females did not develop to the same advanced stages as those of the conspecifically mated females. Sequencing of a 250-bp ITS-2 fragment in eggs that originated from a cross between an R. sanguineus female and an R. turanicus male showed a genotype similar (except by a deletion of 1 thymine) to that observed in the mother, arguing against fertilization by a trans-specific male. These findings suggest that male sex peptides are species-specific molecules that influence both full engorgement and oocyte maturation. Mechanical stimulation of the gonopore alone was insufficient for the completion of the entire process of vitellogenesis.

Made available in DSpace on 2013-09-27T14:53:29Z (GMT). No. of bitstreams: 1 WOS000305307000004.pdf: 224251 bytes, checksum: ad832f37dbd1804429de3fe43307203c (MD5) Previous issue date: 2012-06-01 Made available in DSpace on 2013-09-30T18:47:15Z (GMT). No. of bitstreams: 1 WOS000305307000004.pdf: 224251 bytes, checksum: ad832f37dbd1804429de3fe43307203c (MD5) Previous issue date: 2012-06-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:56:03Z No. of bitstreams: 1 WOS000305307000004.pdf: 224251 bytes, checksum: ad832f37dbd1804429de3fe43307203c (MD5) Made available in DSpace on 2014-05-20T13:56:03Z (GMT). No. of bitstreams: 1 WOS000305307000004.pdf: 224251 bytes, checksum: ad832f37dbd1804429de3fe43307203c (MD5) Previous issue date: 2012-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) 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 semiengorged 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. (C) 2012 Elsevier B.V. All rights reserved. Univ Estadual Paulista, UNESP, Dept Biol, Inst Biociencias, BR-13506900 Rio Claro, SP, Brazil UNESP, Fac Ciencias Agr & Vet, Dept Patol Vet, BR-14884900 Jaboticabal, SP, Brazil Univ Estadual Paulista, UNESP, Dept Biol, Inst Biociencias, BR-13506900 Rio Claro, SP, Brazil UNESP, Fac Ciencias Agr & Vet, Dept Patol Vet, BR-14884900 Jaboticabal, SP, Brazil FAPESP: 07/57809-5 FAPESP: 07/59020-0 FAPESP: 09/54125-3 CNPq: 308733/2006-1