Molecular identification of Cryptosporidium species in Canadian post-weaned calves and adult dairy cattle.

Cryptosporidium is a zoonotic protozoan parasite that is distributed globally and impacts both human and animal health. There are over 40 species of Cryptosporidium described to date, of which four (C. parvum, C. bovis, C. ryanae and C. andersoni) are routinely reported in cattle. The goal of this study was to identify the Cryptosporidium species infecting dairy cattle from across Canada using cow fecal samples and post-weaned calf rectal swabs obtained through the Canadian National Dairy Study. A total of 353 cattle samples (117 pooled rectal fecal swabs from post-weaned calves and 236 cow fecal samples) from 175 herds across the 10 Canadian provinces were analysed by targeting Cryptosporidium's small subunit ribosomal RNA (SSU rRNA or 18S) gene. Herd prevalence of Cryptosporidium was 27.4% nationally, ranging from 0% in Saskatchewan (SK) to 62% in Prince Edward Island (PE). The national prevalence of Cryptosporidium cattle infections was 15.4% in pooled rectal fecal swab samples from post-weaned calves and 16.1% in adult cows. Sanger sequence analysis of the SSU rRNA gene target revealed that C. bovis, C. andersoni and C. ryanae occurred in both adults and post-weaned calves, with C. bovis as the predominant species detected in pooled fecal swab samples of post-weaned calves (9/18, 50%) and C. andersoni as the predominant species in cows (25/38, 66%). Cryptosporidium parvum was not observed in any of the pooled rectal swab samples from post-weaned calves but was observed in one mixed infection of C. bovis/C. parvum in an adult cow. The fifth species identified in this study was C. muris and was present in two adult cows. Low concentrations of oocyst equivalents, as measured by quantitative real-time PCR (qPCR) of the SSU rRNA gene copy number, were observed in a subset of cattle samples. Cryptosporidium andersoni concentrations varied from province to province, with the widest range and highest counts in cows from PE. In conclusion, oocysts from Cryptosporidium species observed in this study are shed into the environment, contributing to the environmental load. However, the Cryptosporidium species in the post-weaned calves and cows found in this study pose a lower risk to the dairy cattle than if they were infected with C. parvum. Similarly, these Cryptosporidium species pose only a small risk to public health as the three species are infrequently reported in humans globally and have not been reported in Canadians to date.