title: Cryptosporidium in farmed animals: the detection of a novel isolate in sheep.
abstract: We describe the discovery of polymorphisms in the Cryptosporidium oocyst wall protein (COWP) gene conferring a novel restriction fragment length polymorphism (RFLP) pattern in 26/60 (43%) isolates from a flock of sheep sampled following a waterborne outbreak of human cryptosporidiosis. The sheep isolates showed identical PCR-RFLP patterns to each other by COWP genotyping but different from those of most currently recognised genotypes, including the major Cryptosporidium parvum genotypes 1 and 2. Sequence analysis of the 550bp amplicon from the COWP gene was compared with a DNA coding region employed in previous studies and showed the novel isolate to differ from other Cryptosporidium species and C. parvum isolates by 7-21%. The sheep-derived isolates were compared at this and further three Cryptosporidium gene loci with isolates from other farmed animals. The loci employed were one in the thrombospondin related adhesive protein (TRAP-C2) gene and two in the 70kDa heat shock protein (HSP70) gene (CPHSP1 and 2). Other animal samples tested in our laboratory were from clinically ill animals and all contained C. parvum genotype 2. The sheep in which the novel isolate was identified were healthy and showed no symptoms of cryptosporidiosis, and the novel sheep isolate could represent a non-pathogenic strain. Our studies suggest that a previously undetected Cryptosporidium sub-type may exist in sheep populations, reflecting the increasingly recognised diversity within the parasite genus.
authors.affiliation: PHLS Cryptosporidium Reference Unit, Swansea PHL, Singleton Hospital, Swansea SA2 8QA, UK. rachel.chalmers@phls.wales.nhs.uk
authors.forename: R M
authors.initials: RM
authors.lastname: Chalmers
authors.forename: K
authors.initials: K
authors.lastname: Elwin
authors.forename: W J
authors.initials: WJ
authors.lastname: Reilly
authors.forename: H
authors.initials: H
authors.lastname: Irvine
authors.forename: A L
authors.initials: AL
authors.lastname: Thomas
authors.forename: P R
authors.initials: PR
authors.lastname: Hunter
mesh_terms: D000818:Animals; D003457:Cryptosporidiosis / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*; D003458:Cryptosporidium / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics*; D004196:Disease Outbreaks; D005243:Feces / Q000469:parasitology; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D015800:Protozoan Proteins / Q000737:chemistry / Q000145:classification / Q000235:genetics; D017422:Sequence Analysis, DNA; D012756:Sheep; D012757:Sheep Diseases / Q000453:epidemiology / Q000469:parasitology*
chemical_list: D016054:DNA, Protozoan; D018840:HSP70 Heat-Shock Proteins; D015800:Protozoan Proteins; C106305:oocyst wall protein, Cryptosporidium; C069428:thrombospondin-related adhesive protein, protozoan