Molecular detection of Cryptosporidium parvum in wild rodents (Phyllotis darwini) inhabiting protected and rural transitional areas in north-central Chile

Abstract

Wild rodents often harbor Cryptosporidium species that can be transmitted to multiple mammal hosts. In Chile, little is known about Cryptosporidium in wild rodents, and available studies have been focused on morphological findings with no molecular-based evidence. A longitudinal survey was conducted between 2021 and 2022 to investigate the occurrence of Cryptosporidium spp. in populations of the Darwin's leaf-eared mouse (Phyllotis darwini) living in protected and rural transitional areas in north-central Chile, using staining and molecular methods. A total of 247 fecal samples were collected and examined by the modified Ziehl–Neelsen (ZN) staining test, 54 of which were positive for Cryptosporidium-like oocysts. Molecular analyses were carried out by PCR of the partial 18S ribosomal RNA and 60 kDa glycoprotein (gp60) genes. Cryptosporidium infection was confirmed in 34 samples (13.7 %) based on the PCR amplification, and individual (i.e., sex, and body mass index) and ecological variables (i.e., type of site and season) were not statistically significant (p > 0.05). Using the nucleotide sequencing of the partial 18S rRNA gene, Cryptosporidium parvum was identified in nine isolates. Also, C. parvum subgenotype family IIa was determined in seven samples by the partial gp60 gene, including the subtype IIaA17G4R1 in two samples. This is the first molecular evidence of Cryptosporidium parvum IIa in Phyllotis darwini in Chile. These results indicate potential cross-species transmition between wild rodents and domestic-wild animals in north-central Chile. More research is needed to understand better the role of wild rodents in the transmission of Cryptosporidium spp. in Chile.