Rhipicephalus (Boophilus) microplus: Distinct acute phase proteins vary during infestations according to the genetic composition of the bovine hosts, Bos taurus and Bos indicus

Tick bites may trigger acute phase responses. Positive and negative acute phase proteins were measured in infested cattle genetically resistant and susceptible to ticks. During heavier infestations levels of haptoglobin increased significantly in susceptible bovines; levels of serum amyloid A increased in resistant bovines; levels of alpha-l-acid glycoprotein decreased significantly in resistant bovines; levels of transferrin decreased significantly in susceptible bovines. In conclusion, tick infestations trigger acute phase responses and enhancement of specific acute phase proteins differs according to the genetic composition of hosts. Acute phase proteins may constitute useful biological signatures for monitoring the stress induced by tick infestations. (c) 2007 Elsevier Inc. All rights reserved.

Mitotic and meiotic chromosomes of a southern Brazilian population of Boophilus microplus (Acari, Ixodidae)

Using conventional staining with acetic orcein and C-banding techniques it was investigated constitutive heterochromatin chromosomal polymorphisms and the mitotic and the meiotic behavior of male and female chromosomes of Boophilus microplus (Canestrini, 1887). Some differences were detected in the population of southern Brazil as compared to the data of other authors for populations in other latitudes. The differences being mainly concerned with the distribution of constitutive centromeric heterochromatin and variation in the length of heterochromatic blocks in the pericentromeric regions of some chromosome pairs.

New histochemical and morphological findings in the female genital tract of Boophilus microplus (Acari, Ixodidae): an attempt toward the elucidation of fertilization in ticks

At present not only is the site of fertilization in ticks still unknown but it is also unclear as to how this mystery can be solved. Signs of fertilization can be observed throughout the female genital tract and these can be clues for the elucidation of the unsolved questions relating to ticks fertilization. In Boophilus microplus (Canestrini, 1887) the most important signs are the following: the final eversion of the acrosomal canal in females ready for oviposition; the presence of small tubules, resembling the subplasmalemal process of the spermatozoon between the oviduct cells; budding nuclei throughout the female genital tract; and the two Feulgen and DAPI positive areas in the oocyte at vitelogenesis. These morphological characteristics suggest that fertilization takes place in the internal cylinder which extends from the uterus to the ovary itself.

Poderá o carrapato transmitir a lepra?: mais quatro amostras de culturas de bacilos acido-alcool resistentes obtidas de carrapatos (2 de "Amblyomma cajennense"e 2 de "Boophilus microplus") infectados em leprosos do Paraná: 3ª nota

The AA. carried out experiments in the leprosarium São Roque, State of Paraná, South Brazil, to verify if the cattle tick Boophilus microplus could be experimentally infected in lepers, which was true. The AA. Tried also to be ascertained if Boophilus microplus and Amblyomma cajennense could change of hosts during their feedings which was true, both ticks continue feeding, the last species for many days, after being transferred from one to another leper. The junior A. describes in full their experiments and also a dermatites caused by tick bites. The senior A. brought to Rio de Janeiro most of the infected ticks for examination, which revealed a very high positivity. He smeared the sediments of lots of both species of ticks in Loewenstein medium and after a variable periode of incubation at 37° C. he obtained four new samples of cultures of acid-fast organisms, two from Amblyomma cajennense and two from Boophilus microplus. These cultures are being studied and will be inoculated into laboratory animals. The senior A. inoculated new batches of white rats with sediments of many ticks infected in lepers. Various hypotheses of both previous notes upon the subject now are verified facts. The A. is accumulating facts to draw the conclusions in the future. He also suggested the leprosy workers in the interior of the country to cooperate with him in such important studies, specially in the habitat of lepers in the rural zones of various States.

Successful vaccination against Boophilus microplus and Babesia bovis using recombinat antigens

Current methods for the control of the cattle tick Boophils microplus and the agent of bovine babesiosis, Babesia bovis are unsatisfactory. Effective immunological control of both parasites would have great advantages. However, naturally acquired immunity to the tick is generally unable to prevent serious production losses. A vaccine against the tick, based on a novel form of immunization, is being developed. A protective antigen has been isolated from the tick, characterized and produced as an effective, recombinant protein. A vaccine incorporating this antigen is currently undergoing field trials. In the Australian situation, improved tick control will probably increase endemic instability with respect to B. bovis. Fortunately, a trivalent, recombinant B. bovis vaccine has also been developed. This too is now undergoing pre-registration field trials.

Infection rate of Babesia spp. sporokinetes in engorged Boophilus microplus from an area of enzootic stability in the State of Minas Gerais, Brazil

The infection rates of Babesia sporokinetes in engorged Boophilus microplus were evaluated during a 2-year period in a dairy farm located in an area of enzootic stability. Every 14 days engorged females were collected from calves and from adult animals. Ticks were incubated at 27 ± 0.5ºC and 80-90% relative humidity and Babesia infection rates were determined by microscopic examination of Giemsa-stained hemolymph smears. After 52 collections, 2105 ticks were obtained, from which 982 were collected from calves and 1123 from cows. The total Babesia infection rate was 10%, however the incidence was higher (p < 0.05) in ticks collected from calves (17.5%) than in those collected from cows (3.6%). Females collected from cows showed the highest infection rates in January, March, and August, and absence of infection in April and May. Ticks feeding on calves were infected throughout the experimental period. The infection rates of engorged females collected from naturally infected calves that were artificially infested with Babesia-free-larvae of B. microplus gradually decreased until the calves were four months old. No differences were observed among infection rates of ticks collected from calves maintained under natural conditions.

Controle de larvas de Boophilus microplus por Metarhizium anisopliae em pastagens infestadas artificialmente

O objetivo deste trabalho foi avaliar a eficiência do controle exercido por Metarhizium anisopliae na população de Boophilus microplus, em pastagens de Brachiaria brizantha, e do híbrido Tifton 85 (Cynodon spp.), artificialmente infestadas com fêmeas ingurgitadas do carrapato. Trinta canteiros com 1 m² de área cada foram distribuídos aleatoriamente. Quinze foram pulverizados com esporos do fungo e quinze controles em cada forrageira, constituindo cinco repetições de cada tratamento, foram infestados com número e peso padronizados de fêmeas ingurgitadas do ácaro. Aplicou-se o fungo, na concentração de 1,8x10(8) conídios mL-1, em três situações: pulverização antes da infestação com o carrapato, após a infestação e posterioriormente à emergência das primeiras larvas nos capins. A ação do fungo foi avaliada no 35º, 38º, 41º, 48º, 55º e 61º dia pós-infestação, por meio da contagem de larvas recuperadas. Obteve-se controle de larvas do ácaro, que, nas avaliações realizadas entre o 35º e o 48º dia pós-infestação, variou entre 87% e 94%. As médias das contagens de estágios larvares do carrapato foram menores em todas as amostragens realizadas no capim-Tifton 85, indicando que houve efeito da pastagem na ação do fungo. A situação de aplicação influencia a atividade do fungo, com melhor resultado nas coletas realizadas entre o 41º e 55º dia após infestação em B. brizantha, e aplicação dos conídios logo após a emergência das primeiras larvas.

Larvicidal potential of Sapindus saponaria to control the cattle tick Boophilus microplus

The objective of this study was to evaluate the larvicidal potential of a crude ethanol extract (CEE) of soapberry Sapindus saponaria stem peel on the cattle tick Boophilus microplus. Tick larvae obtained by incubating engorged females, collected from naturally infested cattle, were placed in envelopes of filter paper impregnated with different concentrations of CEE in the test group, and distilled water in the control group. Four repetitions were made with each solution (n>120). Mortality was observed after 48 hours. Lethal concentration values of 1,258 ppm (LC50) and 6,360 ppm (LC99) were obtained.

Abordagem sobre o controle do carrapato Rhipicephalus (Boophilus) microplus no sul do Rio Grande do Sul

Na região sul do Rio Grande do Sul a infestação dos bovinos por Rhipicephalus (Boophilus) microplus ocorre, principalmente, entre os meses de outubro e abril, devido às condições climáticas. Além do conhecimento do ciclo biológico desse parasito, também é fundamental conhecer a epidemiologia, para estabelecer estratégias de controle. No Rio Grande do Sul, e também no Brasil, existem poucos estudos epidemiológicos a respeito da resistência aos acaricidas. Além disso, a grande área geográfica e a deficiência estrutural quanto ao uso e acesso a bancos de dados dificultam a obtenção de dados confiáveis. O presente estudo teve como objetivo realizar um inquérito abordando a percepção dos produtores da região sul do Rio Grande do Sul, quanto à identificação de populações de R. (B.) microplus difíceis de controlar com acaricidas e os fatores de risco para a seleção de populações de carrapatos resistentes. Para execução do trabalho foram coletados dados sobre o controle do carrapato de bovinos de corte, em 85 propriedades de sete municípios, localizados na região sul do Estado. Os resultados revelaram a existência de associação positiva entre a dificuldade de controlar o carrapato com os acaricidas e o grau de instrução do proprietário (até o ensino fundamental com OR=3,67 e p=0,01) e o número de aplicação de carrapaticida por ano (superior a 4 com OR=4,05 e p=0,006). Esses resultados indicam também que propriedades com mais de 100 bovinos de corte em criação extensiva, na região sul do rio Grande do Sul apresentam características que podem contribuir para uma maior vida útil dos carrapaticidas do que as verificadas em outras regiões do País.

Caracterização do controle de Haematobia irritans e Rhipicephalus (Boophilus) microplus no Triângulo Mineiro e Alto Paranaíba, Minas Gerais

O controle eficaz de Haematobia irritans (mosca-dos-chifres) e Rhipicephalus (Boophilus) microplus (carrapato-do-boi) é um fator crucial à maior rentabilidade da pecuária brasileira, porém ainda constitui-se um desafio. Um maior conhecimento das práticas adotadas no combate destes parasitos faz-se necessário para que se possam estruturar estratégias de controle mais próximas da realidade do produtor rural e mais fáceis de serem executadas. Este estudo caracterizou, através de entrevistas, as práticas adotadas no controle desses ectoparasitos em 23 propriedades da mesorregião do Triângulo Mineiro e Alto Paranaíba, Minas Gerais. A maioria dos entrevistados reconheceu a importância desses parasitos para a atividade pecuária, entretanto, diversos problemas relativos ao controle parasitário foram observados. O controle da mosca-dos-chifres e do carrapato-do-boi era uma prática rotineira em, respectivamente, 17,4% e 95,7% das propriedades, geralmente realizado em função de elevado grau de infestação dos animais. Mais de seis aplicações de ectoparasiticidas eram realizadas por ano em todas as propriedades que empregavam tratamentos para o controle da mosca e em 76,5% daquelas que combatiam o carrapato. Os produtos eram aplicados principalmente com bombas costais manuais (63,6%) e sem a contenção dos animais em 45,5% das propriedades. A diluição dos produtos segundo recomendações dos fabricantes era realizada por 45,4% dos entrevistados, porém, 63,6% aplicavam um volume de solução por animal menor do que o tecnicamente recomendado. Utilizava-se principalmente a associação de piretróides e organofosforados para o controle de ambos os parasitos. Apesar de utilizar rotineiramente ectoparasiticidas, o uso de equipamento de proteção individual (EPI) não era comum entre os entrevistados. A maioria dos entrevistados conhecia algumas características epidemiológicas dos parasitos, entretanto, o controle parasitário adotado na maior parte das propriedades tende a comprometer não apenas a eficácia dos tratamentos, mas a suscetibilidade dos parasitos e a sustentabilidade do controle.

Assessing bovine babesiosis in Rhipicephalus (Boophilus) microplus ticks and 3 to 9-month-old cattle in the middle Magdalena region, Colombia

Babesia sp. is a protozoan hemoparasite that affects livestock worldwide. The Colombian Middle Magdalena is an enzootic region for babesiosis, but there is no previous research providing detail on its transmission cycle. This study aims to assess some Babesia sp. infection indicators in cattle and ticks from the area, by using direct microscopic and molecular techniques to detect the infection. In the cattle, 59.9% and 3.4 % positivity values for B. bigemina and mixed infection (B. bovis + B. bigemina) were found respectively. In ticks, the positivity of B. bigemina reached 79.2% and 9.4% for the mixed infection. The degree of infestation in the region was 3.2 ticks per bovine. There was positive correlation between tick control acaricide frequencies and infestation in bovines. This leads us to infer that control periodicity greater than 90 days, in stable zones, is an abiotic factor that benefits the acquisition of protective immunity in calves, the natural control of the infection and eventual disease absence. It is necessary to monitor the disease by applying new entomological and parasitological indicators showing the complexity of this phenomenon.

Resposta de linfonodos em bovinos inoculados a campo com a vacina recombinante rSBm7462 anti Rhipicephalus (Boophilus) microplus)

A dependência exclusiva de compostos químicos para o controle de Rhipicephalus (Boophilus) microplus tornou-se uma das maiores preocupações científicas e econômicas dos últimos anos, e como consequência, estão sendo realizadas pesquisas para o desenvolvimento de vacinas. O objetivo deste trabalho foi avaliar a resposta de linfonodos de bovinos imunizados a campo com o peptídeo rSBm7462 anti R. (B.) microplus. Foram utilizados 14 bovinos mestiços (Bos taurus x Bos indicus), com idades entre 4-10 meses, mantidos em duas propriedades rurais do norte do estado de Minas Gerais. Os animais receberam três imunizações do peptídeo rSBm7462, aplicados por via subcutânea, com intervalo de 30 dias. Após 15 dias de cada imunização, os linfonodos pré-escapulares foram coletados e fixados por 18 horas em formol. Posteriormente, foram incluídos em Paraplast e as amostras foram coradas pela técnica hematoxilina-eosina (HE) para a observação de eventos celulares. Para a identificação do antígeno nos linfonodos dos animais imunizados, foi realizada a técnica de imuno-histoquímica (IHQ) com o método peroxidase-anti-peroxidase (PAP). A resposta de linfonodos dos bovinos inoculados foi avaliada pelas análises de formação de centros germinais (CG), hiperplasia de cordões medulares (CM) e a presença do antígeno rSBm7462 em células PAP+, demonstrando que o peptídeo recombinante rSBm7462 induz uma resposta imune adaptativa T-dependente, caracterizada nos tecidos linfóides secundários pela formação de estruturas que conferem afinidade e memória imunológica.

Characterization of proteinases from the midgut of Rhipicephalus (Boophilus) microplus involved in the generation of antimicrobial peptides

Background: Hemoglobin is a rich source of biologically active peptides, some of which are potent antimicrobials (hemocidins). A few hemocidins have been purified from the midgut contents of ticks. Nonetheless, how antimicrobials are generated in the tick midgut and their role in immunity is still poorly understood. Here we report, for the first time, the contribution of two midgut proteinases to the generation of hemocidins. Results: An aspartic proteinase, designated BmAP, was isolated from the midgut of Rhipicephalus (Boophilus) microplus using three chromatographic steps. Reverse transcription-quantitative polymerase chain reaction revealed that BmAP is restricted to the midgut. The other enzyme is a previously characterized midgut cathepsin L-like cysteine proteinase designated BmCL1. Substrate specificities of native BmAP and recombinant BmCL1 were mapped using a synthetic combinatorial peptide library and bovine hemoglobin. BmCL1 preferred substrates containing non-polar residues at P2 subsite and polar residues at P1, whereas BmAP hydrolysed substrates containing non-polar amino acids at P1 and P1`. Conclusions: BmAP and BmCL1 generate hemocidins from hemoglobin alpha and beta chains in vitro. We postulate that hemocidins may be important for the control of tick pathogens and midgut flora.

Structure and Mode of Action of Microplusin, a Copper II-chelating Antimicrobial Peptide from the Cattle Tick Rhipicephalus (Boophilus) microplus

Microplusin, a Rhipicephalus (Boophilus) microplus antimicrobial peptide (AMP) is the first fully characterized member of a new family of cysteine-rich AMPs with histidine-rich regions at the N and C termini. In the tick, microplusin belongs to the arsenal of innate defense molecules active against bacteria and fungi. Here we describe the NMR solution structure of microplusin and demonstrate that the protein binds copper II and iron II. Structured as a single alpha-helical globular domain, microplusin consists of five alpha-helices: alpha 1 (residues Gly-9 to Arg-21), alpha 2 (residues Glu-27 to Asn-40), alpha 3 (residues Arg-44 to Thr-54), alpha 4 (residues Leu-57 to Tyr-64), and alpha 5 (residues Asn-67 to Cys-80). The N and C termini are disordered. This structure is unlike any other AMP structures described to date. We also used NMR spectroscopy to map the copper binding region on microplusin. Finally, using the Gram-positive bacteria Micrococcus luteus as a model, we studied of mode of action of microplusin. Microplusin has a bacteriostatic effect and does not permeabilize the bacterial membrane. Because microplusin binds metals, we tested whether this was related to its antimicrobial activity. We found that the bacteriostatic effect of microplusin was fully reversed by supplementation of culture media with copper II but not iron II. We also demonstrated that microplusin affects M. luteus respiration, a copper-dependent process. Thus, we conclude that the antibacterial effect of microplusin is due to its ability to bind and sequester copper II.

(1)H, (15)N and (13)C assignments of the Rhipicephalus (Boophilus) microplus anti-microbial peptide microplusin

Microplusin, a Rhipicephalus (Boophilus) microplus anti-microbial peptide (AMP) is the first member of a new family of cysteine-rich AMPs with histidine-rich regions at the N- and C-termini, which is being fully characterized by biophysical and biochemical methods. Here we report the NMR resonance assignments for (1)H, (15)N, and (13)C nuclei in the backbone and side chains of the microplusin as basis for further studies of structure, backbone dynamics and interactions mapping.