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.

Distribution of Rickettsia rickettsii in ovary cells of Rhipicephalus sanguineus (Latreille1806) (Acari: Ixodidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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

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

Determination of the LC50 of selamectin (active principle of the antiparasitic Revolution (R), Pfizer) applied on engorged female of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Toxicity effect of the acaricide fipronil in semi-engorged females of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae): Preliminary determination of the minimum lethal concentration and LC50

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

Determination of LC50 of permethrin acaricide in semi-engorged females of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae)

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

Esterase profile in a pyrethroid-resistant Brazilian strain of the cattle tick Boophilus microplus (Acari, Ixodidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização da população de ectoparasitos em cães de núcleos de expansão urbana de Juiz de Fora, Minas Gerais, Brasil

No presente trabalho caracterizou-se a população de ectoparasitos em cães de núcleo de expansão urbana de Juiz de Fora, MG. As coletas foram realizadas de julho a setembro de 2003, examinando-se 101 cães SRD (sem raça definida). Os ectoparasitos foram coletados através de inspeção visual e tátil dos animais e acondicionados em frascos com etanol 70°GL e identificados sob estereoscopia. Espécimes foram clarificados e montados para análise em microscopia fotônica. Dentre os sifonápteros, Ctenocephalides felis foi mais prevalente (64,35%) com intensidade média de 6,12 ± 5,37 sifonápteros/cão, seguido por Rhopalopsyllus lutzi (3,96%), híbrida C. felis X C. canis (1,98%), Tunga penetrans (1,98%). Aúnica espécie de fitiráptero encontrada foi Trichodectes canis (7,92%) com 1,3 ± 0,51 fitiráptero/cão. Dentre os ixodídeos, Rhipicephalus sanguineus foi a espécie mais prevalente (49,50%) com intensidade média de 6,44 ± 10,2 ixodídeos/ cão, seguido por Amblyomma cajennense (3,96%), Boophilus microplus (2,97%), A. ovale (1%) e A. aureolatum (1%). As ninfas de ixodídeos foram separadas em ninfas de Amblyomminae (58,41%) com 10,11 ± 10,09 ninfas/cão e ninfas Rhipicephalinae (24,75%) com 2,64 ± 3,25 ninfas/cão. em 3,96 % dos cães foram encontradas larvas de ixodídeo.

Development of an enzyme-linked immunosorbent assay for detection of IgM antibodies to Babesia bigemina in cattle

A crude antigenic preparation of Babesia bigemina was used to develop an ELISA for the detection of IgM antibodies. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkerboard titrations. Negative sera from cattle imported from tick-free areas, serum samples collected from infected B. bigemina cattle were used to validate the test. The specificity was 94% and sensitivity of the Elisa 87.5%. Sera from 385 cattle deriving from areas free from tick-borne diseases, which were submitted to a preimmunization process, were screened by this technique. The Elisa detected seroconversion on the 14th day post-inoculation in animals either infested with Boophilus microplus ticks (infected with B. bigemina), or inoculated with B. bigemina infected blood. Antibody titers decreased after day 33; however, all animals remained positive until the end of the experiment (124 days). The ELISA described may prove to be an appropriate serological test for the detection of IgM antibodies against B. bigemina.

Soroprevalência de Borrelia burgdorferi latu sensu associada à presença de carrapatos em cães de áreas rurais do Estado do Rio de Janeiro, Brasil

Pesquisou-se a presença de anticorpos contra Borrelia burgdorferi latu sensu em cães de áreas rurais de sete municípios do Estado do Rio de Janeiro, pelo ensaio imunoenzimático indireto, associando-se os resultados com a presença de carrapatos nestes animais. de 199 cães examinados, 15,58% estavam positivos, com títulos que variaram de 400 (13,57%) a 1600 (0,5%). Os casos positivos se distribuíram uniformemente nos sete municípios. Não houve diferença estatística (P > 0,05) de soropositivos entre as faixas etárias dos cães acima de seis meses. Carrapatos foram encontrados e coletados em 71 (35,68%) cães, dos quais 24,1% estavam infestados com Amblyomma cajennense, 13,6% com Rhipicephalus sanguineus, 2,5% com Amblyomma aureolatum e 1,5% com Amblyomma ovale. Dos animais soropositivos para B. burgdorferi, 38,7% apresentavam A. cajennense e 22,6% apresentavam R. sanguineus, não havendo, entretanto, correlação positiva entre a presença do carrapato e sorologia positiva.

Análise de fatores genéticos e ambientais que afetam a infestação de fêmeas bovinas da raça Caracu por carrapatos (Boophilus microplus)

Este trabalho foi conduzido com o objetivo de avaliar o efeito de fatores de meio sobre a infestação de bovinos Caracu pelo carrapato Boophilus microplus (Canestrini, 1887) e estimar parâmetros genéticos do grau de infestação por esse ectoparasita. Foram realizadas contagens em fêmeas de dois rebanhos, nas quatro estações, por dois anos consecutivos (setembro/1998 a julho/2000). Contou-se o número de carrapatos (NC) em um dos lados do animal e atribuiu-se escore visual (EC) de acordo com a quantidade de carrapatos no animal. Foram feitas de uma a oito avaliações, totalizando-se 4.079 e 3.994 observações de NC e EC, respectivamente, em 718 animais. Os dados foram analisados pelo método dos quadrados mínimos com um modelo que incluiu efeitos de rebanho (R), cor do animal (C), R x C, animal dentro de R x C como erro a, ano e estação da avaliação, espessura de pelame e idade do animal como covariável. As estimativas dos componentes de variância foram obtidas pelo método da máxima verossimilhança restrita livre de derivadas, utilizando-se um modelo que incluiu os efeitos fixos de grupo de contemporâneos (fazenda-ano-época), espessura do pelame e idade do animal como covariável e os efeitos aleatórios aditivos diretos e de ambiente permanente. Antes das análises, a variável NC foi transformada para log10 (n + 1) e EC para (x + 0,5)½, em que n é o número de carrapatos contados no animal e x, o escore (0 a 4). A incidência de carrapatos foi maior no verão e, quanto maior a espessura do pelame, maior o nível de infestação. As estimativas de herdabilidade e repetibilidade foram, respectivamente, 0,22 e 0,29 para NC e 0,15 e 0,21 para EC; a correlação genética entre NC e EC foi igual a 1,00. Os resultados sugerem que é possível obter progresso genético para resistência a carrapato pela seleção.

Tests to determine LC50 and discriminating doses for macrocyclic lactones against the cattle tick, Boophilus microplus

Laboratory tests were carried out on larvae and adults of the cattle tick Boophilus microplus to determine the toxicity of macrocyclic lactone acaricides (MLs). Technical and commercial MLs were used in larval packet test (LPT), larval immersion test (LIT) and adult immersion test (AIT). Tn LIT and AIT the toxicity of MLs was much higher than for LPT. In the AIT, diluting the injectable formulation of MLs in water was as effective as dilution in ethanol + Triton X-100. LC50, LC99.9 and 95% confidence limits were determined so that a discriminating dose (DD) could be set for larval and adult tests in order to diagnose potential resistance to MLs in field samples of the tick. These DDs are for Australian strains of B. microplus and may not be suitable for other strains until further work is carried out.The value of these diagnostic tests can only be verified if or when resistance to MLs emerges in ticks. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Histopathology of tick-bite lesions in naturally infested capybaras (Hydrochoerus hydrochaeris) in Brazil

In the present work features of tick-bite lesions were evaluated in capybaras naturally infested with Amblyomma cajennense and Amblyomma dubitatum ticks. Gross appearance of tick bite site was characterized by a mild swelling and erythema. Microscopic examination revealed the cement cone, a tube-like homogenous eosinophilic mass penetrating deep into the dermis. This structure was surrounded in the dermis by a cellular infiltrate and free eosinophilic granules and was associated to edema of variable intensity. Necrosis was a common feature deep in the dermis particularly at the far end of the eosinophilic tube. Hyperplasia, cellular edema and occasionally necrosis of keratinocytes could be seen at both sides of the ruptured epidermis. Cellular infiltrate was constituted overwhelmingly by polymorphonuclear leukocytes with eosinophilic granules. In capybaras cells with such features can be either eosinophils or heterophils (pseudoeosinophils), the latter being the equivalent of neutrophils of other mammals. Ultrastructural analysis of the cellular infiltrate revealed the predominance of heterophils over eosinophils. Mononuclear cells and mast cells and, in lesser numbers, basophils were also seen at skin attachment sites. The presence of heterophils in the reaction of capybaras against Amblyomma ticks is an outstanding feature but its role in the reaction to the tick is not known. It is however speculated that capybara heterophils might be associated with a more permissive environment for tick feeding and pathogen transmission as already shown for the equivalent cell type, the neutrophil, in the reaction of the dog against the Rhipicephalus sanguineus tick.

Life cycle and host specificity of Amblyomma triste (Acari : Ixodidae) under laboratory conditions

We report biological data of two generations of Amblyomma triste in laboratory and compared the suitability of different host species. Infestations by larval and nymphal stages were performed on guinea pigs (Cavia porcellus), chickens (Gallus gallus), rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus), wild mice (Calomys callosus), dogs (Canis familiaris) and capybaras (Hydrochaeris hydrochaeris). Infestations by adult ticks were performed on dogs, capybaras and rabbits. Tick developmental periods were observed in an incubator at 27degreesC and RH 90%. Guinea pigs were the most suitable hosts for larvae and nymphs, followed by chickens. The remaining host species were less suitable for immature ticks as fewer engorged ticks were recovered from them. Mean larval feeding periods varied from 3.8 to 4.7 d between different host species. Mean larval premolt periods ranged from 8.9 to 10.4 d. Nymphal mean feeding periods varied from 4.2 to 6.2 d for ticks fed on different host species. Premolt period of male nymphs (mean: 15.4 d) was significantly longer than that of female nymphs (14.7 d). Female nymphs were significantly heavier than male nymphs. The overall sex ratio of the adult ticks emerged from nymphs was 0.9:1 (M:F). Capybaras were the most suitable host for the tick adult stage as significantly more engorged females were recovered from them and these females were significantly heavier than those recovered from dogs or rabbits. The life cycle of A. triste in laboratory could be completed in an average period of 155 d. The potential role of guinea pigs, birds and capybaras, as hosts for A. triste in nature, is discussed.

CROSS-REACTIVITY BETWEEN HARD TICK ANTIGENS

1. The present study was carried out to determine the target cells and tissues for anti-tick immunoglobulins using an indirect immunohistochemical technique.2. Sections in triplicate prepared from unfed ticks Rhipicephalus appendiculatus, R. evertsi and Amblyomma variegatum were used to assess the cross-reactivity of serum from guineapigs naturally infested with these tick species or immunized against them.3. The sections showed slight (+) to strong (++++) labelling of several structures in the tick body, e.g. salivary gland, gut lumen and malpighian tubules, depending on the serum used.4. The immune serum resulting from the immunization of guinea pigs with an extract of unfed nymphs of R. appendiculatus ticks showed the most intense cross-reactivity with the sections examined.