First detection of Cryptosporidium scrofarum, an agent with zoonotic potential, in wild boars (Sus scrofa) in the Americas

Abstract

Cryptosporidiosis is a zoonosis with worldwide distribution and is important because it presents a risk to One Health, despite being neglected by the World Health Organization (WHO). Therefore, it was investigated the occurrence of Cryptosporidium spp. in 44 free-ranging wild boars slaughtered in the northwest region of the state of São Paulo, Brazil. A fecal sample from each animal was collected directly from the rectal ampulla after slaughter. The parasitological investigation was carried out using the malachite green negative staining technique for the detection of Cryptosporidium spp. oocysts. An aliquot of all stool samples was subjected to genomic DNA extraction using the GenElute™ Stool DNA Isolation kit and the Polymerase Chain Reaction (PCR) was performed, amplifying the fragments of the 18S rRNA subunit gene, to the molecular detection of Cryptosporidium spp. Samples positive in the 18S gene for the last agent mentioned above were also subjected to amplification of specific fragments of the heat shock protein HSP70. To verify the association between the positivity and negativity of Cryptosporidium spp. and the variables analyzed (sex and age group), statistical analysis was performed using Fisher's Exact Test. In this research, it was not observed infectious evolutionary forms of the protozoan mentioned in the coproparasitological exams. However, it was detected Cryptosporidium spp. in 6.8 % (3/44) of the animals investigated, by amplification of the 18S rRNA and HSP 70 genes, and by genetic sequencing, two samples with 100 % similarity to Cryptosporidium scrofarum were verified. Therefore, it was detected this species for the first time in free-ranging wild boars in the Americas.