Navegando por Palavras-chave "Rhipicephalus Microplus"

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    Estudo Funcional Da Proteína Rmsei Presente No Intestino Do Carrapato Rhipicephalus Microplus E Sua Participação Na Infecção Por Anaplasma Marginale
    (Universidade Federal de São Paulo (UNIFESP), 2017-03-29) Gomes, Cicera Maria [UNIFESP]; Tanaka, Aparecida Sadae [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Ticks are important vectors of diseases to humans and other vertebrates. The Rhipicephalus (Boophilus) microplus is an exclusive ectoparasite of cattle, being responsible for transmission of Anaplasma sp and Babesia sp, etiologic agents of the disease called "tick fever". The R. microplus causes great damage to livestock, resulting in considerable losses in the production of meat, milk and leather. The ectoparasite control using acaricides is the main method, however it can lead to bovine contamination, as well as the environment. An altemative method to tick control is the production of vaccine, however, to date no vaccine appeared effective for Brazilian cattle. In attempt to identify important proteins for R. microplus, which may compose a multi-vaccine antigens, the aims of this work are: to study an inhibitor of human neutrophil elastase presents in the B. microplus saliva (BmSEI). RmSEI The recombinant protein was produced by cloning into the vector pET26b expression in cells and BL21 pLysS (DE3). The purified rRmSEI was valued about different serine proteases. However, the rRmSEI not presented inhibitory activity as different enzymes tested. For a conventional PCR analysis showed que transcript of RmSEI be presente in all tissue analyzed: salivary gland, fat body, gut, and hemocytes, ovary, with lower expression ofthe transcript in hemocytes. The Analysis of the presence of protein in all tissues, demonstrated a presence in all tissues analyzed with prevalence in the ovary. Preliminary results of gene silencing experiment in gut and ovarian by interference RNA showed 61% silencing RmSEI in gut and a 50% reduction of the microbiota composition; 49.1% of silencing the expression RmSEI in ovarian and a reduction of36.7% ofmicrobiota present in ovariano In tick embryonic cellline BME26, and infected with A. marginale, demonstrated that RmSEI gene silencing in BME26 cells line not change the number of bacteria A. marginale in tum the bacteria A. marginale reduces relative expression RmSEI in BME26 cells line. Thus we conclude that although bacteria infection reduce expression RmSEI, its reduction by RNA interference does not promote bacterial growth, suggesting that RmSEI is not a direct effector, but part of the immune tick response, probably within a system wherein other molecules are involved.
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    Identificação de potenciais antígenos do carrapato Rhipicephalus (Boophilus) microplus para o desenvolvimento de vacinas
    (Universidade Federal de São Paulo (UNIFESP), 2020-07-30) Silva, Fernando Allan Abreu [UNIFESP]; Tanaka, Aparecida Sadae [UNIFESP]; Universidade Federal de São Paulo
    The tick Rhipicephalus (Boophilus) microplus is responsible for considerable economic losses to the livestock sector. The main control method of infestation of this parasite is by acaricides and can result in the selection of resistant ticks, environment and products contamination, between other factors. The vaccine is a more sustainable alternative, but the products available on the market are not efficient in the Brazilian territory. Although the tick genome has not yet been elucidated, transcriptomes and proteomes can indicant potential targets for selection of antigens. Two proteins sequences were selected for this work, a sphingomyelinase D – like and a cathepsin A – like from a tick transcriptome. The DNA sequences that codified these two proteins were cloned into the pET14b expression vector and confirmed by DNA sequencing. The analysis of the translated amino acids of cathepsin A – like showed the absence of amino acids residues important for the activity of this enzyme when compared to others cathepsins A sequences. However, it was not possible to confirm experimentally this data since the recombinant protein was not express in bacterial system. In contrast, sphingomyelinase D – like was expressed in bacteria in large quantities, but in the form of inclusion body. The sphingomyelinase D – like was partially purified in the presence of urea. The analysis of sphingomyelinase D – like primary structure showed a replacement of a histidine for an asparagine in an important position for the enzymatic activity. Therefore, the perspective of this work is to obtain the purified sphingomyelinase D – like and carry out its biochemical and functional characterization.