First identification of Cryptosporidium meleagridis in wild white-eyed parakeet (Psittacara leucophthalmus).

In this research, we investigated the genetic diversity of Cryptosporidium spp. in 65 free-living white-eyed parakeets (Psittacara leucophthalmus) from the central-eastern region of São Paulo state, Brazil. Using real-time nested PCR of the 18S rRNA gene and conventional nested PCR of the actin and heat shock protein (HSP-70) genes, two samples were positive for Cryptosporidium spp., representing an prevalence of 3.08% (95% CI: 0.85-10.54%). However, oocysts of this protozoan were not observed through coproparasitological examination, nor were clinical signs characteristic of cryptosporidiosis noted in any bird. The amplicons obtained in this study were subjected to bidirectional Sanger sequencing. Among the positive samples, one presented sequences of sufficient quality for analysis of the 18S rRNA and HSP70 genes, exhibiting 100% and 99.64% genetic similarity, respectively, with Cryptosporidium meleagridis sequences deposited in GenBank. Phylogenetic analysis using HSP-70 gene sequences from Cryptosporidium spp. confirmed that the amplified Cryptosporidium sequence is closely related to C. meleagridis sequences, grouping within the same clade. Nested PCR targeting the parasite surface glycoprotein gene (GP60) was performed; however, a sufficient number of amplicons were not obtained, making subgenotyping impossible. We believe this to be the first report of C. meleagridis, a protozoan with zoonotic potential in the synanthropic white-eyed parakeets (P. leucophthalmus).