title: Molecular characterisation of Theileria orientalis in imported and native bovines from Pakistan.
abstract: The epidemiological aspects of Theileria orientalis in Pakistan are unknown; therefore, investigations using sensitive and precise molecular techniques are required. This study reports the first molecular characterisation of T. orientalis detected from imported (Bos taurus) and native cattle (Bos indicus×Bos taurus) and buffaloes (Bubalus bubalis) selected from four districts of Punjab, Pakistan. DNA samples from blood (n=246) were extracted and tested using conventional PCR utilising the major piroplasm surface protein (MPSP) gene and multiplexed tandem PCR (MT-PCR). Theileria orientalis DNA was detected (15%; 22/147) only in imported cattle by conventional PCR, whereas 24.5% (36/147), 6% (3/50) and 6.1% (3/49) of the imported cattle and native Pakistani cattle and buffaloes, respectively were test-positive for T. orientalis using MT-PCR. Using MT-PCR, the prevalence of T. orientalis was significantly higher (P<0.0001) in imported cattle compared to that of detected in native Pakistani bovines. The prevalence of T. orientalis and DNA copies of chitose and ikeda were significantly higher (P<0.05) in imported cattle than those detected in native Pakistani bovines. DNA sequencing of amplicons of the conventional PCR revealed the presence of buffeli, chitose and ikeda genotypes of T. orientalis. Phylogenetic analysis revealed that the MPSP sequences of buffeli, chitose and ikeda from imported cattle were closely related to those sequences reported previously from Australia and other regions. This study provides the first survey of T. orientalis infection in imported and native bovines in Pakistan, and highlights the need for future studies to understand the spread of transboundary animal diseases.
authors.affiliation: Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Werribee, Victoria, Australia. Electronic address: hgebremikael@student.unimelb.edu.au.
authors.forename: Hagos
authors.initials: H
authors.lastname: Gebrekidan
authors.affiliation: University College of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Bahawalpur, Punjab, Pakistan.
authors.forename: Tariq
authors.initials: T
authors.lastname: Abbas
authors.affiliation: Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Punjab, Pakistan.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Wajid
authors.affiliation: Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering, Faisalabad, Punjab, Pakistan.
authors.forename: Aamir
authors.initials: A
authors.lastname: Ali
authors.affiliation: Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Werribee, Victoria, Australia.
authors.forename: Robin B
authors.initials: RB
authors.lastname: Gasser
authors.affiliation: Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Werribee, Victoria, Australia. Electronic address: jabbara@unimelb.edu.au.
authors.forename: Abdul
authors.initials: A
authors.lastname: Jabbar
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics; D001315:Australia; D002417:Cattle; D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics; D058889:Molecular Typing; D010154:Pakistan; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics; D016794:Theileria / Q000145:classification* / Q000235:genetics*; D013801:Theileriasis / Q000469:parasitology*
chemical_list: D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D015800:Protozoan Proteins; C115173:major piroplasm surface protein, Theileria
keywords: Australia; Buffalo; Cattle; MT-PCR; PCR; Pakistan; Theileria orientalis