A Multi-locus Study for Detection of Cryptosporidium Species Isolated from Calves Population, Liverpool; UK

Cryptosporidium is an obligate intracellular protozoan parasite infecting a wide range of hosts. The current study investigated the genetic profile of Cryptosporidium species in calves in Liverpool, England. Fifty-two calve fecal samples were collected from a farm and initially screened by Auramine phenol, modified Ziehl-Neelsen and ELISA. PCR analysis of 18S rRNA gene was carried out for the positive samples. Then, positive PCR samples were genotyped by an 18S rRNA- based PCR-RFLP, COWP - based PCR- RFLP; PCR of GP60 and HSP70 genes. Additionally, sequence analysis was carried out based on representative isolates of four loci. Cryptosporidium oocysts and antigens were detected in 34 out if 52 (65.4%) samples using screening techniques. Genotype analysis showed the presence of C. hominis and C. parvum in one and thirteen samples, respectively. Furthermore, subtypes of C. hominis Ib, C. parvum IIa; C. parvum subtype 2 were identified by GP60 and HSP70 sequences, respectively. These findings indicate the diversity of the molecular characteristics of Cryptosporidium species in calves’ isolates. Moreover, referring to the literature; we report two new subtypes of C. parvum IIa and a rare case of C. hominis Ib in calves population.