Free living amoeba diversity on Monjolinho River at State of São Paulo - morphological and molecular approaches

• Grant number: 18/20693-4
• Support Opportunities: BIOTA-FAPESP Program - Regular Research Grants
• Start date: February 01, 2019
• End date: January 31, 2022
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Otavio Henrique Thiemann
• Grantee: Otavio Henrique Thiemann
• Host Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
• Associated researchers: Douglas Cedrim Oliveira ; Jacob Lorenzo-Morales ; María Reyes Batlle ; Odete Rocha

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 key, as well as 18S rDNA sequencing. 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 FLA biodiversity and establishing protocols to that intent, besides to bringing important public health information. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

BELLINI, NATALIA KARLA; MOREIRA DA FONSECA, ANA LETICIA; REYES-BATLLE, MARIA; LORENZO-MORALES, JACOB; ROCHA, ODETE; THIEMANN, OTAVIO HENRIQUE. Isolation of Naegleria spp. from a Brazilian Water Source. PATHOGENS, v. 9, n. 2, FEB 2020. (18/20693-4)

BELLINI, NATALIA KARLA; THIEMANN, OTAVIO HENRIQUE; REYES-BATLLE, MARIA; LORENZO-MORALES, JACOB; COSTA, ADRIANA OLIVEIRA. A history of over 40 years of potentially pathogenic free-living amoeba studies in Brazil - a systematic review. Memórias do Instituto Oswaldo Cruz, v. 117, p. 16-pg., 2022-01-01. (18/20693-4)