Immunomodulatory effects of tick saliva

Ticks are bloodsucking ectoparasites that cause great damage to host organisms, so these ectoparasites are of great importance in medicine and veterinary medicine. All the biological success achieved by ticks is due to the action of bioactive components present in their saliva, which are synthesized by the salivary glands. These components have great diversity of functions such as enabling feeding and the permanence of ectoparasites on hosts, since they modulate immune system acting as complement inhibitors, immunosuppressors, cytokine expression modulator and chemokine binders of hosts. In addition, these components are an important source of protective antigens. In this sense, salivary glands/saliva are considered a potential source of multifunctional molecules. In this context, many studies have been conducted aiming at searching to establish a better understanding on the biology and morphophysiology of some organs such as salivary glands, as well as elucidate the complex relationship of these ectoparasites with their hosts. Such studies are conducted with the main objective of developing new immunobiological products aimed at the alternative control of ticks, as well as for the identification and isolation of bioactive molecules with pharmacological properties and with great therapeutic potential in the search for treatments for some diseases.

Effect of Amblyomma cajennense Ticks on the Immune Response of BALB/c Mice and Horses

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Antigens from Rhipicephalus sanguineus ticks elicit potent cell-mediated immune responses in resistant but not in susceptible animals

In the present study we compared the immunological reactions between Rhipicephalus sanguineus tick-infested susceptible (dogs and mice) and tick-resistant hosts (guinea pigs), elucidating some of the components of efficient protective responses against ticks. We found that T-cells from guinea pigs infested with adult ticks proliferate vigorously in the presence of concanavalin A (ConA), whereas ConA-induced cell proliferation of tick-infested mice and dogs was significantly decreased at 43.1 and 94.0%, respectively, compared to non-infested controls. Moreover, cells from mice and dogs submitted to one or three successive infestations did not exhibit a T-cell proliferative response to tick antigens, whilst cells from thrice tick-infested guinea pigs, when cultured with either a tick extract or tick saliva, displayed a significant increase in cell proliferation. Also, we evaluated the response of tick-infested mice to a cutaneous hypersensitivity test induced by a tick extract. Tick-infested mice developed a significant immediate reaction, whereby a 29.9% increase in the footpad thickness was observed. No delayed-type hypersensitivity (DTH) reaction was detected. Finally, the differential cell count at the tick attachment site in repeatedly infested mice exhibited a 6.6- and 4.1-fold increase in the percentage of eosinophils and neutrophils, respectively, compared to non-infested animals, while a decrease of 77.0-40.9 in the percentage of mononuclear cells was observed. The results of the cutaneous hypersensitivity test and the cellular counts at the tick feeding site for mice support the view that tick-infested mice develop an immune response to R. sanguineus ticks very similar to dogs, the natural host of this species of tick, but very different from guinea pigs (resistant host), which develop a DTH reaction in addition to a basophil and mononuclear cell infiltration at the tick-attachment site. In conclusion, saliva introduced during tick infestations reduces the ability of a susceptible animal host to respond to tick antigens that could stimulate a protective immune response. As a consequence, the animals present a lack of DTH response and disturbed cellular migration to tick feeding site, which can represent a deficient response against ticks. © 2003 Elsevier B.V. All rights reserved.

Study of caffeine urine and saliva of horses subjected to urinary acidification

The study of caffeine in racing horses has been of growing concern in veterinary sports medicine since the Association of Racing Commissioners International (ARCI) stated that it has no valid therapeutic use in racehorses. We examined the kinetic alterations in the urinary excretion and salivary secretion of caffeine in seven horses subjected to urinary acidification using ascorbic acid because this procedure can simulate the acidosis that follows anaerobic exercise. They participated in two treatment groups: the control group (SG) received 500 ml of saline and then 2.0 mg kg(-1) caffeine i.v. 30 min later; and the acidified group (AG) was subjected to urinary acidification with ascorbic acid at a dose of 0.5 g kg(-1) i.v. and then 2.0. mg kg(-1) caffeine i.v. 30 min later. Samples were collected 30 min before caffeine administration, immediately before caffeine administration (time zero) and at 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, 48 and 72 h afterwards. The samples were assayed by gas chromatography. The mean urinary pH for SG was 8.2, but for AG it was as low as 5.9 at 4 h, extending acidosis for up to 8 h. The kinetic curves for the two groups were similar for urinary excretion and salivary secretion. Differences occurred only in peak excretion and peak secretion in SG obtained at 1 h and 30 min, respectively, and in AG at 2 h and 1 h, respectively. This could be explained, in part, to the diuresis in AG compared with SG, resulting in less concentrated urine in the former group. The large difference between the pK(a) of caffeine and the pH of the medium may be responsible for the similar pharmacokinetics observed for the two groups. Copyright (C) 2004 John Wiley Sons, Ltd.

Microscopic features of tick-bite lesions in anteaters and armadillos - Emas National Park and the Pantanal region of Brazil

The naturally occurring wildlife host associations between ticks and tick-borne pathogens found in the neotropics are poorly described. Understanding tick-bite lesions is important as these are the site of host reaction to and pathogen delivery by ticks. As part of a comprehensive study concerning established and emerging tick-host relationships. The present work describes some aspects of tick-bite lesions in anteaters and armadillos captured at the Emas National Park and the Pantanal region of Brazil. Biopsies were of skin were taken and examine. Tick feeding sites of all animals displayed an eosinophilic homogeneous mass, the cement cone, and, occasionally, a feeding cavity underneath the tick attachment site. At these locations the epidermis was usually thickened due to keratinocyte hyperplasia. The main dermal changes included tissue infiltration with a varying number of inflammatory cells, edema, hemorrhage. and vascular dilatation. Cellular infiltration of the dermis was predominantly composed of mononuclear cells, neutrophils. and eosinophils. Mast cells were also seen in both non-parasitized and parasitized skin but were found in higher numbers at perivascular sites and in parasitized skin. Basophils were not seen at tick attachment sites of anteaters or armadillos.

Sequential histopathology at the Rhipicephalus sanguineus tick feeding site on dogs and guinea pigs

The tick Rhipicephalus sanguineus is a very common parasite of dogs worldwide. Dogs seem unable to acquire resistance against this tick species, whereas guinea pigs demonstrate a very strong resistance following primary infestation. We studied the inflammatory reaction at the R. sanguineus tick feeding site on dogs and guinea pigs during primary and tertiary infestations at different time intervals after attachment. Biopsies were collected after 4, 24, 48 and 96 hours. Changes that were found in all experimental groups included a cone of cement around the mouthparts of the tick, epidermal hyperplasia, edema and inflammatory cell infiltration in the dermis directly underneath the tick attachment site. Dogs reacted to ticks mainly with neutrophils, particularly after repeated exposure. Mast cells and mononuclear leukocytes were also present. Guinea pigs reacted to R. sanguineus mainly with mononuclear cells, eosinophils and basophils. These cells were particularly numerous after repeated exposure to R. sanguineus. Our results suggest that basophils and eosinophils are involved in resistance of guinea pigs to R. sanguineus and that neutrophils in dogs have little effect against this tick species.

Antibody and blood leukocyte response in Rhipicephalus sanguineus (Latreille, 1806) tick-infested dogs and guinea pigs

The dog is considered to be the natural host of Rhipicephalus sanguineus and is unable to develop appreciable resistance even after repeated feedings. The guinea pig develops strong resistance after one infestation with adult ticks. Antibody (IgG) titres against tick salivary gland antigens (SGAs) and blood leukocyte numbers in dogs and guinea pigs undergoing experimental R. sanguineus tick infestations were measured to detect a possible correlation with susceptibility or resistance of hosts. Since infested dogs develop an immediate hypersensitivity reaction to R. sanguineus antigens, total and anti-R. sanguineus SGA IgE levels were also measured in this host species. IgG and IgE antibody levels were determined by an enzyme-linked immunosorbent assay (ELISA) along three consecutive infestations of both hosts. Most dogs and guinea pigs displayed low IgG levels against R. sanguineus SGAs, though marked differences in individual response were observed. Although dog's total serum IgE levels increased significantly after infestations, no change in the amount of anti-salivary gland IgE was detected. Total and differential blood cell counts were determined in dogs and guinea pigs during primary and secondary infestation. In dogs, a tertiary infestation and a subsequent higher infestation level were also evaluated. Infested dogs did not display any alteration in blood leukocyte counts throughout the experiment. Guinea pigs, on the other hand, developed a significant basophilia during primary infestation which increased further during secondary infestation. These data reveal similarities and differences in the reactions of resistant and non-resistant hosts to ticks. They contribute for the understanding of such host-parasite relationships and will hopefully aid in the development of immune control of ticks. (C) 2003 Elsevier B.V. All rights reserved.

Survey for Tick-Borne Zoonoses in the State of Espirito Santo, Southeastern Brazil

Blood samples collected from 201 humans, 92 dogs, and 27 horses in the state of Espirito Santo, Brazil, were tested by polymerase chain reaction, indirect immunofluorescence assays, and indirect enzyme-linked immunosorbent assay for tick-borne diseases (rickettsiosis, ehrlichiosis, anaplasmosis, borreliosis, babesiosis). Our results indicated that the surveyed counties are endemic for spotted fever group rickettsiosis because sera from 70 (34.8%) humans, 7 (7.6%) dogs, and 7 (25.9%) horses were reactive to at least one of the six Rickettsia species tested. Although there was evidence of ehrlichiosis (Ehrlichia canis) and babesiosis (Babesia cams vogeli, Theileria equi) in domestic animals, no human was positive for babesiosis and only four individuals were serologically positive for E. canis. Borrelia burgdorferi-serologic reactive sera were rare among humans and horses, but encompassed 51% of the canine samples, suggesting that dogs and their ticks can be part of the epidemiological cycle of the causative agent of the Brazilian zoonosis, named Baggio-Yoshinari Syndrome.

Antigen-presenting cells in draining lymph nodes of goats repeatedly infested by the Cayenne tick Amblyomma cajennense nymphs

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular detection of tick-borne bacterial agents in Brazilian and exotic captive carnivores

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Guinea Pigs Develop Cutaneous Basophilia after Repeated Infestations by Nymphs of the Tick Amblyomma triste

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of Metarhizium anisopliae fungus on off-host Rhipicephalus (Boophilus) microplus from tick-infested pasture under cattle grazing in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cellular and molecular characterization of an embryonic cell line (BME26) from the tick Rhipicephalus (Boophilus) microplus

The cellular and molecular characteristics of a cell line (BME26) derived from embryos of the cattle tick Rhipicephalus (Boophilus) microplus were studied. The cells contained glycogen inclusions, numerous mitochondria, and vesicles with heterogeneous electron densities dispersed throughout the cytoplasm. Vesicles contained lipids and sequestered palladium meso-porphyrin (Pd-mP) and rhodamine-hemoglobin, suggesting their involvement in the autophagic and endocytic pathways. The cells phagocytosed yeast and expressed genes encoding the antimicrobial peptides (microplusin and defensin). A cDNA library was made and 898 unique mRNA sequences were obtained. Among them, 556 sequences were not significantly similar to any sequence found in public databases. Annotation using Gene Ontology revealed transcripts related to several different functional classes. We identified transcripts involved in immune response such as ferritin, serine proteases, protease inhibitors,. antimicrobial peptides, heat shock protein, glutathione S-transferase, peroxidase, and NADPH oxidase. BME26 cells transfected with a plasmid carrying a red fluorescent protein reporter gene (DsRed2) transiently expressed DsRed2 for up to 5 weeks. We conclude that BME26 can be used to experimentally analyze diverse biological processes that occur in R. (B.) microplus such as the innate immune response to tick-borne pathogens. (C) 2008 Elsevier Ltd. All rights reserved.

Resistance of cattle of various genetic groups to the tick Rhipicephalus microplus and the relationship with coat traits

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)