Project Details


Name Diversidade de amebas de vida livre no Rio Monjolinho no estado de São Paulo - abordagens morfológicas e moleculares   FAPESP Project
Website http://www.ifsc.usp.br/~biota_fla
Validity Date 2018 - 2021
Team Otavio Henrique Thiemann - Coordenador - Instituto de Física de São Carlos/IFSC/USP Douglas Cedrim Oliveira - Pesquisador Associado - Campus Rio Verde/IFGRV/IFG Jacob Lorenzo-Morales - Pesquisador Associado - Universidad de La Laguna/ULL María Reyes Batlle - Pesquisador Associado - Universidad de La Laguna/ULL Odete Rocha - Pesquisador Associado - Universidade Federal de São Carlos/UFSCAR
Participating Institutions Instituto de Física de São Carlos/IFSC/USP Campus Rio Verde/IFGRV/IFG Universidad de La Laguna/ULL Universidade Federal de São Carlos/UFSCAR
Biome The Monjolinho river is the sampling site. Geographical position of each sampling sites A to E are: A (22° 0ʹ 2.87ʹʹ S, 47° 50ʹ 8.96ʹʹ W) B (21° 58ʹ 40.6884ʹʹ S, 47° 52ʹ 25.50ʹʹ W) C (21° 59ʹ 45.40ʹʹ S, 47° 54ʹ 6.32ʹʹ W) D (22° 0ʹ 52.41ʹʹ S, 47° 55ʹ 26.65ʹʹ W) E (22° 1ʹ 34.92ʹʹ S, 47° 57ʹ 26.29ʹʹ W).
Abstract Naegleria, Acanthamoeba, Sappinia and Balamuthia comprise the most studied genera of the Free Living Amoeba (FLA) group, whose protists have a worldwide distribution being recovered from either soil or freshwater samples. They are capable of causing serious and fatal infections in humans. However, little is known about the biodiversity of FLA in general and of the incidence of the pathogenic FLA in Brazil. Thereby, this research aims to identify the environmental occurrence of FLA in five sites of a public freshwater supply source, called Monjolinho River at São Carlos – São Paulo, Brazil. The methodology adopted to address this goal includes the water limnological characterization, the morphological and molecular analysis of the protists present in the samples. The latter will be performed using a set of 18S rDNA primers for global (FLA group) sequencing by a high throughput strategy in an Illumina HiSeq technology. The amoeba present in the water samples will be cultured and the isolated trophozoites will be observed with scanning electron microscopy. Four different staining methods will be applied and observed by optical microscopy for morphological identification with the Page’s taxonomic key, as well as 18S rDNA sequencing and phylogenetic reconstruction. The project has advanced by establishing water collection methodology, DNA extraction and PCR amplification protocols. Test samples have been cultured at 26 ºC, 37 ºC and 44 ºC aiming at the identification of thermotolerant species. This proposal is in line with the BIOTA-FAPESP initiative to evaluate the microbial biodiversity, FLA in specific, and establishing protocols to that intent, besides to bringing important public health information.
Expected Products Identification of free-living amoebas present in the water environment.
Keywords
Occurrences 5
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