Molecular Diagnostic studies and Phylogenetic analysis of Theileria species in Egypt

Amira A. T. AL-Hosary1 , Laila S. Ahmed1 , Ulrike Seitzer2

Infectious Diseases, Department of Animal Medicine, Faculty of Veterinary Medicine, Assiut University, Assiut71526, Egypt. 2Veterinary Infection Biology and Immunology, Department of Veterinary Cell Biology and Immunology, Research Center,Borstel, Parkallee 22, 23845, Borstel-Germany The corresponding author: Amira Adel TahaAL-Hosary Faculty of Veterinary Medicine (Department of Animal Medicine, Infectious Diseases), Assiut University, Egypt,

Mobil phone: (002)01004477501.Fax.(088)2366503,(088)2354130 E-mail,amiraelhosary@yahoo.com

Abstract :

Bovine theileriosis is a destructive disease affect the livestock production in Egypt. In the current study 210 cattle and 37 buffaloes from different localities in Upper Egypt were collected and tested by Tams1 target based polymerase chain reaction (Tams1 PCR) and Reverse Line Blot (RLB) assay that target 18S rRNA gene of Theileria spp.flowed by sequencing and phylogenetic analysis for identification of theileria species in both cattle and buffaloes. Tams-1 PCR confirmed the infection in 46.19% and 8.11% while RLB assay confirmed the infection in 65.24% and 51.35%in cattle and buffaloes, respectively. RLB was used as reference test to evaluateTams-1 PCR assay as a diagnostic tool used for diagnosis of bovine theileriosis, the sensitivity of this test was (70.80% and 15.79%) in cattle and buffaloes, respectively while its specificity was 100% in both animal’s species. The PCR products were hybridized on Biodyne C membrane which containing the spacer-oligonucleotide sequences of Theileria spp. Phylogenetic analysis was carried out by using neighbor joining (NJ) method. The obtained results concluded that Assiut and AL-Fayoum governorates have different species of the theileriaprotozoan parasite include Theileriaannulata, Theilerialestoquardi,Theileriauilenbergi and Theileriaovis. On the other hand in ALWadi AL-Jadid governorate Theileriaannulatais the only species founded. All partial sequences were deposited in the GenBankTM databases and available under accession numbers as followingT. annulata (GenBankTM: KF781307, GenBankTM: KF922216&GenBank TM:KF922219), T. lestoquardi (GenBankTM:KF771185), T. ovis (GenBankTM:KF922218) and T. uilenbergi (GenBankTM:KF781308, GenBankTM:KF771184, GenBank TM: KF922215 &GenBank TM: KF922217). Key words Tams-1 PCR, RLB,18S rRNA, Neighbor joining.