Biodiversity of Blastocystis spp. from human and dogs fecal samples from the six Brazilian biomes: parasitological and molecular characterization

• Grant number: 12/05388-4
• Support Opportunities: Regular Research Grants
• Start date: July 01, 2012
• End date: December 31, 2014
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Jeffrey Jon Shaw
• Grantee: Jeffrey Jon Shaw
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Antonio Francisco Malheiros ; Antonio Humberto Hamad Minervino ; Lucile Maria Floeter-Winter ; Raimundo Alves Barrêto Júnior ; Ronaldo Cesar Borges Cryschek

Abstract

Blastocystis spp. is a protist commonly diagnosed in human and animal's feces such as dogs, monkeys, pig, cattle, marsupials, other mammals and birds, verifying its importance as a zoonotic agent. Several studies worldwide have elucidated the etiology of the parasite in humans as in animals, through subtyping. In contrast, in Brazil there is a need for more comprehensive studies applied to public health that show the molecular parasitological diagnosis, the actual specific etiologic agent may have an important role in the epidemiology of this parasite. For this, we will collect fecal samples of humans and other animals of the six Brazilian biomes: Amazon, Caatinga, Cerrado, Atlantic Forest, Pampa, Pantanal, visiting a total of 24 municipalities (four cities geographically distributed selected in each biome. In each of these cities 100 samples will be collected from human and 100 from dogs, totaling 4800 fecal samples (2400 to 2400 of human and dog). The human samples will be collected from patients treated by Hospitals and Regional Health Programs Family, while the fecal samples of dogs will be collected from animals from the Zoonosis Control Centers or veterinary clinics. The samples will be preserved in 96% ethanol and kept under refrigeration at 4 ° C until analysis by three microscopic techniques (Direct examination; Hoffmann, Ritchie). In the positive samples in at least one of the tests, will be performed the Polymerase Chain Reaction (PCR), prior to DNA extraction, for Blastocystis spp., followed by the DNA sequencing in an automatic sequencer of the positive samples. All sequences obtained in the BLAST analysis will be submitted to verify homology with corresponding sequences available in GenBank, and thus perform the genetic identification of the amplified products in the samples tested. (AU)