Molecular Screening for Sarcocystidae in Muscles of Wild Birds from Brazil Suggests a Plethora of Intermediate Hosts for

The genus and the species are the most prevalent sarcocystid organisms found in birds. Molecular phylogenies based on the first internal transcribed spacer of the ribosomal coding DNA (ITS1) have been widely used to identify them. Here, pectoral muscles from 400 wild birds from Brazil were screened by means of molecular methods using nested PCR, and Sanger sequencing yielded amplicons. A pan-sarcocystid ITS1-directed nested PCR revealed 28 birds infected by (ten Piciformes, eight Psittaciformes, five Columbiformes, two Accipitriformes, one Anseriformes, one Passeriformes and one Strigiformes); one infected by (one Accipitriformes); nine infected by unknown or undescribed (six Passeriformes, one Piciformes, one Cathartiformes and one Cuculiformes); and six harboring DNA (three Pelecaniformes, two Falconiformes and one Columbiformes). Samples harboring -related ITS1 sequences were further characterized by means of PCR and sequencing of genetic sequences of three surface antigen coding genes (SAGs). From this, 10 new allelic combinations of SAGs (SAG2, SAG3 and SAG4) were identified, in addition to 11 SAG allelic combinations already found in Brazil. Samples with -unrelated ITS1 sequences were further characterized by means of PCR and sequencing of cytochrome oxidase subunit I coding sequences (CO1) and 18S ribosomal DNA gene (18S rDNA). This study was the first extensive survey of wild birds in Brazil for Sarcocystidae species. It provides the first molecular evidence of natural infection in 14 species, including in the order Piciformes, and shows the high genetic diversity of in intermediate hosts in South America. Evidence of occurrence of at least three non-described species of was also presented in this study. This survey corroborated the ubiquity of infection but revealed surprisingly low prevalence of this parasite (1.5%).