Aplicações da genética na conservação e combate ao tráfico de animais silvestres

Abstract

Different approaches in the area of genetics are able to help species conservation. One is through the identification of wild animals mantained illegally in captivity. Recently the CONAMA resolution No. 487/2018 made the genotyping mandatory in breeding facilities to verify the affiliation of some species, being Saltator similis the species with the highest priority due to the demand for trafficking. Thus, this work had as main focus the development of a paternity test for the species based on microsatellites (STR) using the technology of massively parallel DNA sequencing both for the development and for the analysis of the markers sequences. The paternity test was able to identify the parents even when less than ten loci were used, indicating that the loci developed in this work are very informative and are able to compose the genotyping panel, in addition to the fact that they can also be used in management projects and conservation. Additionally, the complete mtDNA of the species has also been sequenced and annotated, and may assist investigations of crimes against the species when the use of COI alone is not informative enough and also in evolutionary studies. Another approach is synthetic biology, which can help maintain species by reducing the risk of extinction and even creating proxies of extinct species that maintain the ecological function in the ecosystem. One of these pioneering de-extinction projects, in which part of this work was carried out, is the creation of a hybrid of Asian elephant (template species) and woolly mammoth that maintains the ecological functions of the extinct one. For this, it is necessary to know the genomes of both species and perform gene editing. sgRNAs were designed to knockout Asian elephant genes and primers to confirm these edits, as well as FISH protocol standardization for a better understanding of the nuclear structure of woolly mammoth. At the same time, this type of project usually helps in the preservation of template species. In the case of the Asian elephant, EEHV infections have been a complicating factor in its conservation. Little is known about the pathogenicity mechanisms of this virus and possible treatments because its cultivation in the laboratory has not yet been possible. In order to make these studies possible, in this work EEHV fragments were amplified and extracted for later construction of the virus genetic material that could be used in experiments.