Molecular Characterization of Paramphistomes fromCattle from Matebeleland Region (Zimbabwe) using RandomAmplified Polymorphic DNA (RAPDs) and Amplified Ribosomal DNA Restriction Analysis (ARDRA)

Abstract

Molecular Characterization of Paramphistomes fromCattle from Matebeleland Region (Zimbabwe) using RandomAmplified Polymorphic DNA (RAPDs) and Amplified Ribosomal DNA Restriction Analysis (ARDRA) is a published article of the Applied Biology and Biochemistry department.,Paramphistome isolates collected from local abattoirs were genetically characterised using the Random Ampl41i47fied Polymorphic DNA (RAPD) technique and Amplified Ribosomal DNA Restriction Analysis (ARDRA). These isolates were morphologically characterized using median sectioning and five putative species were identified. Of the 18 isolates that were being investigated, 16 were positively identified: three belonged to Calicophoron calicophorum, two were Calicophoron microbothrium, one was Gigantocotyle symmeri, 6 were identified as Calicophoron raja and the other 6 were identified as Calicophoron clavula. A restriction digest of the amplified ITS-2 region of all isolates was done using two restriction enzymes Hae III and Sau 3A1 and the fragments obtained did not show any detectable polymorphisms on all isolates. A total number of 110 bands were generated by RAPD-PCR and 91.82% of these were polymorphic with an average genetic distance of 0.4810+/- 0.185 that showed substantial variability among the paramphistome isolates. The RAPD-PCR technique however, gave banding patterns that on analysis were able to cluster (on the dendrogram) the isolates into their respective species groups and even aid in identifying the two isolates that were not positively identified morphologically as Calicophoron raja. A fragment of approximately 1300bp was generated from primer OPB 07 on Calicophoron microbothrium isolates which can be used as a selectable marker for this species. The findings of the present study therefore showed that the RAPD- PCR technique can be used for molecular identification of paramphistomes