title: Molecular detection of Babesia ovis in sheep and ticks using the gene encoding B. ovis surface protein D (BoSPD).
abstract: The gene encoding Babesia ovis surface protein D (BoSPD) was cloned from B. ovis cDNA library. This gene encodes a polypeptide chain of 155 amino acids, including a predicted 22 amino acid signal peptide. Sequence analysis of the BoSPD suggested that it is a surface protein with no known domains. BLAST analysis followed by multiple alignments showed four orthologs from other Apicomplexan species and suggested that BoSPD is specific for B. ovis. BoSPD-based PCR was then developed to specifically detect B. ovis in experimentally-infected sheep and Rhipicephalus bursa ticks, as well as in field samples. The PCR enabled detection of B. ovis at a calculated parasitemia of 0.0016% and was shown to be specific for B. ovis. Moreover, the BoSPD PCR allowed detection of prolonged subclinical infection in experimentally-infected lambs and in dissected organs of experimentally-infected ticks. Finally, the PCR was used to detect parasitemia in blood samples from naturally-infected sheep and in R. bursa ticks collected from sheep in an infected flock. These results suggest that the BoSPD gene sequence can be used as a specific and sensitive marker, allowing detection of subclinical parasitemia in sheep and in ticks. Based on its predicted properties, BoSPD may be considered as a candidate for anti-B. ovis vaccine development or a target for anti-B.ovis treatment.
authors.affiliation: Division of Parasitology, Kimron Veterinary Institute, PO Box 12, Bet Dagan 50250, Israel. Electronic address: oran.erster@gmail.com.
authors.forename: Oran
authors.initials: O
authors.lastname: Erster
authors.affiliation: Division of Parasitology, Kimron Veterinary Institute, PO Box 12, Bet Dagan 50250, Israel.
authors.forename: Asael
authors.initials: A
authors.lastname: Roth
authors.affiliation: Division of Parasitology, Kimron Veterinary Institute, PO Box 12, Bet Dagan 50250, Israel.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Wolkomirsky
authors.affiliation: Division of Parasitology, Kimron Veterinary Institute, PO Box 12, Bet Dagan 50250, Israel.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Leibovich
authors.affiliation: Division of Parasitology, Kimron Veterinary Institute, PO Box 12, Bet Dagan 50250, Israel.
authors.forename: Igor
authors.initials: I
authors.lastname: Savitzky
authors.affiliation: Israeli Veterinary Field Services, Bet Dagan 50250, Israel.
authors.forename: Shmuel
authors.initials: S
authors.lastname: Zamir
authors.affiliation: Division of Parasitology, Kimron Veterinary Institute, PO Box 12, Bet Dagan 50250, Israel.
authors.forename: Thea
authors.initials: T
authors.lastname: Molad
authors.affiliation: Division of Parasitology, Kimron Veterinary Institute, PO Box 12, Bet Dagan 50250, Israel.
authors.forename: Varda
authors.initials: V
authors.lastname: Shkap
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001403:Babesia / Q000235:genetics* / Q000502:physiology; D001404:Babesiosis / Q000097:blood*; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D018512:Parasitemia / Q000097:blood / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000662:veterinary; D048494:Rhipicephalus / Q000469:parasitology*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D017385:Sequence Homology; D012756:Sheep; D012757:Sheep Diseases / Q000097:blood*
chemical_list: D008565:Membrane Proteins
keywords: Babesia ovis; BoSPD PCR; Domain prediction; Rhipicephalus bursa