Transmiss?o experimental de Rickettsia parkeri para Rhipicephalus sanguineus e Amblyomma sculptum por meio da alimenta??o artificial

• Author(s): Valim, Jaqueline Rodrigues de Almeida
• Date: 2021
• Origin: Oasisbr
• Subject(s): Ixod?deos; Alimenta??o artificial; Caracteriza??o; Infec??o; Ixodidae; Artificial feeding; Molecular characterization; Infection; Medicina Veterin?ria
• Document Type: Doctoral thesis
• Language: Portuguese

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES, Brasil)

Artificial feeding techniques of hematophagous arthropods constitute an important tool for making possible the analysis of biological aspects and the evaluation of the efficiency of arthropods as vectors of bioagents. Plastic tips are a concrete alternative, as it makes it possible to tevaluate the relationship between vectors and pathogens, biological aspects, to test acaricides and to minimize the use of animals in scientific studies. The objectives of this study were to standardize the of plastic tips on the artificial feeding of Amblyomma sculptum, to carry out the experimental study of Rickettsia parkeri for A. sculptum and Rhipicephalus sanguineus by feeding and characterization of proteins in the interaction of this Rickettsia with the in addition to analyzing the egg mass and the rhythm of posture of the non-parasitic phase of the females of experimentally infected ixodids. For infestations rabbits were used young of the species Oryctolagus cuniculus. On the ninth day of infestation in rabbits, females partially of A. sculptum were collected and separated into two bands of initial weight and submitted to artificial feeding for the times of 6, 12 and 24 hours. After standardization, the range used for A. sculptum was 55-150 mg for a period of 12 hours. For the ixod?deo R. sanguineus, the weight range and the time for the infection had previously been standardized, adopted the weight range of 36-80 mg for a period of 12 hours. After infection by feeding artificial females were dissected through the method of exhaustion, the organs ovaries, gland salivary gut and intestine were separated for characterization of proteins using the SDSPAGE technique. The biological parameters in the standardization were not influenced by artificial feeding with the device, for the first time specimens of A. sculptum parthenogens presented complete engorgement when fed in vitro, in the case of the experimental infection by R. parkeri, the two species of ixodids presented alterations in the egg mass and in the posture / oviposition, showing a decrease in quantity and posture peaks were the fourth day for R. sanguineus and the seventh day for A. sculptum. These results will contribute to the studies involving the transmission of pathogens in vitro, as well as minimizing the use of animals in animal experimentation. In the protein profile, bands were found in the polyacrylamide with weight standards ranging from 40kDa to 170kDa.

As t?cnicas de alimenta??o artificial de artr?podes hemat?fagos constituem em uma importante ferramenta por possibilitar a an?lise dos aspectos biol?gicos e a avalia??o da efici?ncia dos artr?podes como vetores de bioagentes. Ponteiras pl?stica consituem-se em alternativa concreta, pois possibilita avaliar a rela??o entre vetores e agentes patog?nicos, aspectos biol?gicos, testar acaricidas e minimizar a utiliza??o de animais em estudos cient?ficos. Os objetivos deste trabalho foram padronizar a utiliza??o de ponteiras pl?sticas na alimenta??o artificial de Amblyomma sculptum, realizar a transmiss?o experimental de Rickettsia parkeri para A. sculptum e Rhipicephalus sanguineus por meio da alimenta??o artificial e proceder a caracteriza??o de prote?nas na intera??o desta Rickettsia com os respectivos ixod?deos, al?m de analisar a massa de ovos e o ritmo de postura da fase n?o parasit?ria das f?meas dos ixodideos infectadas experimentalmente. Para as infesta??es, foram utilizados coelhos jovens da esp?cie Oryctolagus cuniculus. No nono dia de infesta??o nos coelhos, as f?meas parcialmente ingurgitadas de A. sculptum foram coletadas e separadas em duas faixas de peso inicial e submetidas ? alimenta??o artificial pelos tempos de 6, 12 e 24 horas. Ap?s a padroniza??o, a faixa utilizada para A. sculptum foi 55-150 mg por um per?odo de 12 horas. Para o ixod?deo R. sanguineus, a faixa de peso e o tempo de alimenta??o utilizada para a infec??o j? haviam sido padronizados anteriormente, sendo adotada a faixa de peso de 36-80 mg por um per?odo de 12 horas. Ap?s a infec??o pela alimenta??o artificial as f?meas foram dissecadas atrav?s do m?todo do esgotamento, os ?rg?os ov?rios, gl?ndula salivar e intestino foram separados para a caracteriza??o de prote?nas utilizando a t?cnica de SDS-PAGE. Os par?metros biol?gicos na padroniza??o n?o sofreram influ?ncia da alimenta??o artificial com o dispositivo, pela primeira vez exemplares de parten?ginas de A. sculptum apresentaram ingurgitamento completo quando alimentadas in vitro, em se tratando da infec??o experimental por R. parkeri as duas esp?cies de ixod?deos apresentaram altera??o na massa de ovos e no tempo de postura/oviposi??o, apresentando diminui??o na quantidade e os picos de postura foram o quarto dia para R. sanguineus e o s?timo dia para A. sculptum. Estes resultados contribuir?o para o aprofundamento nos estudos envolvendo a transmiss?o de pat?genos in vitro, bem como minimizar a utiliza??o de animais na experimenta??o animal. No perfil prot?ico foram encontradas bandas nos g?is de poliacrilamida com padr?es de peso variando de 40kDa at? 170kDa.