Matias Pablo Juan Szabó

Ticks (Acari: Ixodidae) associated with wild animals in the Pantanal region of Brazil.

Abstract This paper describes the identification of ticks from wild animals of the Pantanal region in Brazil as part of a comprehensive study about established and emerging tick-host relationships and related pathological aspects. Eighty-one animals were captured (representing 13 species, six orders), and ticks were found on 63 (78%). Tick species identified included Boophilus microplus (Canestrini), Amblyomma cajennense (F.), Amblyomma parvum Aragão, Amblyomma pseudoconcolor Aragão, Amblyomma scalpturatum Neumann, Amblyomma nodosum Neumann, Amblyomma ovale Koch, and Amblyomma tigrinum Koch. Dragging from grasslands yielded negative results compared with the high concentration of ticks that were collected from leaves in the forests.

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.

Gene discovery in Boophilus microplus, the cattle tick: the transcriptomes of ovaries, salivary glands, and hemocytes.

Abstract: The quest for new control strategies for ticks can profit from high throughput genomics. In order to identify genes that are involved in oogenesis and development, in defense, and in hematophagy, the transcriptomes of ovaries, hemocytes, and salivary glands from rapidly ingurgitating females, and of salivary glands from males of Boophilus microplus were PCR amplified, and the expressed sequence tags (EST) of random clones were mass sequenced. So far, more than 1,344 EST have been generated for these tissues, with approximately 30% novelty, depending on the the tissue studied. To date approximately 760 nucleotide sequences from B. microplus are deposited in the NCBI database. Mass sequencing of partial cDNAs of parasite genes can build up this scant database and rapidly generate a large quantity of useful information about potential targets for immunobiological or chemical control.

Report on ticks collected in the Southeast and Mid-West regions of Brazil: analyzing the potential transmission of tick-borne pathogens to man

Specimens of ticks were collected in 1993, 1996, 1997, and 1998, mostly from wild and domestic animals in the Southeast and Mid-West regions of Brazil. Nine species of Amblyommidae were identified: Anocentor nitens, Amblyomma cajennense, Amblyomma ovale, Amblyomma fulvum, Amblyomma striatum, Amblyomma rotundatum, Boophilus microplus, Boophilus annulatus, and Rhipicephalus sanguineus. The potential of these tick species as transmitters of pathogens to man was analyzed. A Flaviviridade Flavivirus was isolated from Amblyomma cajennense specimens collected from a sick capybara (Hydrochaeris hydrochaeris). Amblyomma cajennense is the main transmitter of Rickettsia rickettsii (=R. rickettsi), the causative agent of spotted fever in Brazil. Wild mammals, mainly capybaras and deer, infested by ticks and living in close contact with cattle, horses and dogs, offer the risk of transmission of wild zoonosis to these domestic animals and to man.

Immunisation of dogs and guinea pigs against Rhipicephalus sanguineus ticks using gut extract.

Tick-bite naive guinea pigs were inoculated three times with Rhipicephalus sanguineus gut or salivary gland extracts and saponin as adjuvant. Dogs were inoculated three times with gut extract only as this fraction induced a more efficient resistance in guinea pigs (lower tick recovery and lower engorged female weights). Freunds adjuvant and saponin were used as adjuvants for the immunisation of dogs. Freunds adjuvant was used to enhance cellular immunity. The highest level of resistance in dogs was induced by the immunisation with gut extract and Freunds adjuvant. Many female ticks from dogs immunised this way engorged fully but died prior to oviposition. Resistant guinea pigs and dogs seemed to trigger different immune mechanisms against R. sanguineus ticks as damage to parasites also differed. A major role for cellular immunity in the resistance of dogs against R. sanguineus ticks is suggested. Resistance mechanisms against R. sanguineus ticks is discussed.

Skin Test and Tick Immune Status in Susceptible and Resistant Cattle in Brazil

Abstract: A cutaneous hypersensitivity test (CHT) was used to correlate host resistance to ticks and type of reaction elicited to unfed larval extract‐ULE of the cattle tick Boophilus microplus in European and Indian cattle. Twenty calves were separated into four groups of five animals each: naïve or preinfested Indian or European cattle. CHT was induced by intradermal inoculation of 0.1 ml of ULE cattle tick B. microplus (50 μg protein) in the calf ear. Ear thickness was measured using calipers before and 10 min, 1, 2, 6, 18, 24, 48, 72, 96, and 144 h postinoculation (PI). Preinfested European calves showed only an immediate type reaction with maximum response (75% increase in ear thickness) at 10 min PI. On the other hand, preinfested Indian calves presented an immediate response with maximum reaction (70% increase in ear thickness) between 10 min and one hour PI, and a delayed type reaction at 72 h PI (60% increase in ear thickness). These results point out the crucial role of the cellular immune response of cattle in the expression of resistance to cattle tick B. microplus. Skin test might be useful in the ranking of cattle according to the susceptibility/resistance to ticks.

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 dogs 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.

Ticks Associated with Wild Animals in the Nhecolândia Pantanal, Brazil

Abstract: A study of ticks associated with wild animals was carried out from September 1996 to April 1998 at the Fazenda Alegria (21,000 ha), in the Nhecolândia Pantanal, State of Mato Grosso do Sul, Brazil, a sunken plain bordering the upper Paraguay river, located 19 × 08′S; 56 × 46′W. A total of 81 wild animals (13 species, 6 orders) were captured with the aid of nets, and ticks were found on 63 (78%). Tick species identified included Boophilus microplus (Canestrini), Amblyomma cajennense (F.), A. parvum (Aragão), A. pseudoconcolor (Aragão), A. scalpturatum (Neumann), A. nodosum (Neumann), A. ovale (Koch), and A. tigrinum (Koch). Dragging from grasslands (campos) yielded negative results compared to the high concentration of ticks, mainly nymphs, that were collected from leaves in the forests (capão). Predominance of immature instars (Amblyomma genera) was observed in the end of winter (August‐September). Ticks were associated mainly with coatis, deer (Mazama gouazoubira) and anteater, and these animals may play a role in the epidemiology of tick‐transmitted pathogens in the Pantanal if one considers their co‐existence with local domestic animals.

Colonização e lesão em fêmeas ingurgitadas do carrapato Rhipicephalus sanguineus causadas pelo fungo Metarhizium anisopliae

The objective of this work was to verify the penetration of the fungus Metarhizium anisopliae [METSCH. (SOROKIN, 1883)], in to the Rhipicephalus sanguineus ticks (LATREILLE, 1806), as well as the lesions caused in the tissues of the acarus. The form of adherence and penetration of the fungus in the deep tissue was studied through scanning microscopy and the fungus action on the internal tissues was evaluated in conventional hystological sections. In order to evaluate these events, experimental infestations were conducted in 11 engorged females groups of the tick R. sanguineus, containing 12 engorged females in each. Aiming this goal the engorged females were submited to a bath in a conidial suspension in a concentration of 108 conidia/mL during three minutes under manual agitation. In case of control groups the bath was done only in the suspension vehicle. Ticks were processed for histological and scanning microscopy surveys at different intervals after the infection: 1 and 18h and one, two, three, four, five, six, seven, eight, nine and eleven days. It has been observed that the major part of the conidians have germinated within 18h after the inoculation and that the fungus the penetrated the acarus body, trough it´s tegument 48h after the infection. After the penetration the fungus colonized the tickx92s body in a diffuse manner, without any preference for a specific tissue. Among the lesions caused by the fungus, rupture of the tickx92s intestinal wall and the dispersion of the fluid into the hemocel were the most outstanding features. Death of the host occurred 96h pos-infection and the sporulation of the pathogen over the dead acarus began 120 to 144h pos-infection. This work aims to contribute to the establishement of techniques for biological control of the ticks by fungus as an ecological alternative for acaricides.

Histopathology of Rhipicephalus sanguineus (Acari: Ixodidae) ticks fed on resistant hosts

Histological features of Rhipicephalus sanguineus ticks fed on dog, a non resistant host, and on guinea pig, a resistant host, were compared. Unfed ticks and ticks from each host species were collected during first and third infestation and processed for histology. Many ticks from guinea pigs, especially during third infestation, were unattached, dehydrated and small. Only the midgut of ticks fed on guinea pigs had host leukocytes. Vacuolization of midgut cells was observed in all ticks, with exception of those fed on dogs for more than 96xa0h. Ticks of guinea pigs, particularly from third infestation, had vacuolated tracheae and swelling of malpighian tubules. Solely ticks from third infestation of guinea pigs displayed vacuolization of oocytes. Ticks fed on guinea pigs also had an increased number of guanine spherules. Observed alterations in ticks from guinea pigs are discussed.