Diagnosis, risk factors analysis and first molecular characterization of Cryptosporidium spp. in horses from Rio de Janeiro, Brazil

Abstract

An analysis was made of the frequency of Cryptosporidium spp. in fecal samples from horses raised on farms in the Teresópolis city, state of Rio de Janeiro, Brazil, and the risk factors that favored this infection. Between 2019 and 2020, 314 samples of equine feces were collected, 287 of which came from English Thoroughbred horses and 27 from ponies. Information on the horses and their management were retrieved from a stud book and forms filled out by trainers. The fecal samples were subjected to macroscopic analysis, modified Sheather's and Lutz parasitological techniques, safranin staining, and to enzyme-linked immunosorbent assay (ELISA) for the detection of coproantigens. All the samples that tested positive by these techniques underwent partial sequence analysis of the 18S rRNA gene to characterize the protozoan species. Cryptosporidium spp. was identified in 35 (11.1%) of the samples, 34 from English Thoroughbred horses and one from a pony. Based on a logistic regression model, it was found that the presence of dogs and small ruminants on the farms, and drinking water from a spring, were significantly associated with the animals' infection by the protozoan (p < 0.05). Eight of the English Thoroughbred horse samples underwent molecular characterization, which revealed the presence of Cryptosporidium felis in one sample and Cryptosporidium parvum in seven. The seven samples containing C. parvum were subjected to gp60 gene analysis, based on which nucleotide sequences typical of the IIa family were identified, which are usually transmitted from animals to humans. In addition, the genotype IIaA15G2R1, which is considered to have the highest profile of zoonotic transmissibility, was identified in one Thoroughbred horse. This is the first study conducted in the state of Rio de Janeiro that molecularly characterized Cryptosporidium spp. in horses, and the first on the American continent to detect C. felis in the feces of these animals.