Theileria annulata: Genetic characterization of Spanish isolates by isoenzyme electrophoresis and random amplified polymorphic DNA

Isoenzyme electrophoresis and RAPD techniques were used to study the geneti c polymorphism of different Spanish isolates of Theileria annulata in the s chizont and piroplasm stages. Enzyme activity attributable to the parasite was detected in only 5 of the 13 loci studied with isoenzyme electrophoresi s. Of these, differences between the cell lines studied were found only in the loci GPI, ICD, and RI. Only 6 of the 11 primers used in the RAPD genera ted reproducible genomic DNA fingerprints. None of the amplification produc ts generated using primers ILO 509, ILO 525, ILO 872, and no 875 hybridized with DNA of the bovine cell line not infected by T. annulata, BL20, indica ting that this technique can be used with either piroplasm DNA or DNA from parasite schizonts after first passing it through DEAE cellulose columns. T he results obtained with both characterization techniques demonstrate a mod erate degree of polymorphism among the Spanish isolates of this protozoan. (C) 1999 Academic Press.