Free Life Amoebas: development of the visualization tool PVT and identification of Naegleria spp. and Acanthamoeba spp. on the Monjolinho River in São Carlos - SP

• Grant number: 21/04364-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2021
• End date: June 30, 2022
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Otavio Henrique Thiemann
• Grantee: Matheus Issa
• Host Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil

Abstract

Free-living amoebae (AVL) are protist microorganisms widely found in nature, but which, although most of them live freely in the environment, present some optional opportunistic parasitic genera, such as Naegleria spp. and Acanthamoeba spp .. They usually cause infections associated with the Central Nervous System (CNS) and their rapid progression leads patients to death. In Brazil, studies of these microorganisms are quite scarce, with no precise mapping of their environmental distribution, which makes the lack of knowledge about presence of these amoebas is a high risk for the population. In addition, the morphological analyzes performed today are not optimized, making it difficult to process the researched samples efficiently. Therefore, this project aims to develop a visualization program based on taxonomic keys, based on the classification guide Page (1988), and to verify, at the molecular level, the existence of species of the AVL genera mentioned in 5 collection sites of the Monjolinho River in the city of São Carlos, SP. For the first question, the morphological characteristics of the different amoebae described in the guide for the creation of the PVT software (in English, Page's Visualization Tool) are gathered in tables. For the second, traditional molecular biology techniques are used, starting with DNA extractiongenomics of the samples, going through PCR amplification with specific oligonucleotides forgender, amplified DNA sequencing and bioinformatics analysis, performing the alignment with NCBI's BLAST tool from the GenBank database. Thus, it is possible to build phylogenies capable of providing a general environmental overview of the dispersion of these microorganisms in the region, contributing, together with the PVT, to the optimization and complementation of future research on the topic.