Prevalence and molecular characterization of Cryptosporidium spp. in dairy cattle in South Bohemia, the Czech Republic

Abstract

A total of 995 samples from slaughtered dairy cattle (6 months and older) were collected during two consecutive years (from 2007 to 2008), stained by aniline-carbol-methyl violet and examined microscopically. DNA was extracted from Cryptosporidium positive samples and from 200 randomly selected microscopically negative samples. Nested PCR was performed to amplify the partial SSU rRNA and GP60 genes of Cryptosporidium that were subsequently digested by SspI, VspI and MboII restriction enzymes to determine the Cryptosporidium species and genotype present. The highest prevalence of infection (18.2%) was in the animals in age group of 12-18 months. The sequence analyses of PCR-positive specimens identified 41 samples as Cryptosporidium andersoni (4.1%), 2 samples as Cryptosporidium bovis (0.2%), and 1 sample as Cryptosporidium parvum (0.1%). C. bovis was detected only in a group of cattle older than 18 months and C. parvum in heifer which was older than 14 months. Seasonal dependency of Cryptosporidium spp. prevalence was not proven to any significant degree. Infection intensity was generally low. Sequence analysis of the glycoprotein (GP60) gene showed that detected C. parvum belonged to the IIaA16G1R1 subtype. This is the first published report about subtyping of the Cryptosporidium GP60 gene from cattle in the Czech Republic.