Diagnóstico molecular de bioagentes em carrapatos, oriundos de fragmentos do bioma Mata Atlântica, Brasil

Abstract

This work is divided into two chapters and aims to: 1) Identify the species of ticks in the freeliving phase, which occur in the Conservation Units (UC's) in the states of Rio de Janeiro, Espírito Santo (ES) and Minas Gerais, Brazil, and to detect deoxyribonucleic acids (DNA) from bacteria of the genera Rickettsia and Borrelia through polymerase chain reaction (PCR) and 2) Detect molecular evidence of the presence of bacteria and protozoa of the genera Rickettsia, Borrelia , Babesia, Anaplasma and Ehrlichia in ticks collected from wild birds in the state of Rio de Janeiro. In the first study, 17 UC's were visited (national, state and municipal) where free-living ticks were captured using CO2 trap and manual collection. A total of 1,356 ticks were collected, including 1201 larvae, 106 nymphs and 49 adults. The nymphs were identified as Amblyomma spp. and adults such as A. oblongoguttatum, A. sculptum, A. brasiliense, A. dubitatum, A. aureolatum. The samples were submitted to DNA extraction by the phenolchloroform method with precipitation in isopropyl alcohol. In PCR using a partial sequence of the gltA gene as a target, Rickettsia spp. in 10.1% (19/188) of the samples, for Borrelia spp. a partial sequence of the flaB gene was used as a target and no sample was positive. Sequencing of 19 samples identified the presence of Rickettsia bellii DNA. For Chapter II, 33 birds of 20 different species were captured, of which 14 were parasitized by tick nymphs that were identified as A. longirostre. The presence of Rickettsia spp. in three samples, one of which was also positive for Borrelia spp. After sequencing, the co-infected sample showed 100% identity with Rickettsia bellii strain H3 and 91% with Borrelia turcica, two samples had 100% identity with Rickettsia sp. Aranha strain and AL strain.