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

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

The distribution and variation of Ixodid ticks in Matabeleland region Zimbabwe

Ticks have been known to cause severe economic losses due to cattle deaths, low milk production, poor beef quality and poor hide quality. To inform control strategies, it is necessary to carry out tick surveys to determine tick species present in Matabeleland as well as determine their distribution in the region. A total of 119 cattle were sampled from seven sites in   Matabeleland communal and commercial areas, that is, Mangwe, Grills farm, Umguza, Beitbridge, Insiza, Gwanda and Nkayi. Phenotypic identification was done by microscopy and total DNA extraction was performed using the tick legs, which was then followed by Random Amplified Polymorphic DNA (RAPD) analysis to determine genetic variation and DNA polymorphism within the tick population. Of the cattle sampled, 268 ticks were collected and 11 tick species were identified, that is, Amblyomma hebraeum 54%, Hyalomma  truncatum 11%,  Rhipicephalus evertsi evertsi 12%, Rhipicephalus appendiculatus 7.4%, Rhipicephalus (Boophilus) decoloratus 6%, Hyalomma rufipes 3.7%,  Rhipicephalus zambeziensis 1.7%,  Rhipicephalus sanguineus 1.1%,  Rhipicephalus simus 1.1%,  Rhipicephalus lunulatus 0.7% and Amblyomma variegatum 0.7 %. The occurrence of Amblyomma variegatum in this part of the country was a unique finding as the tick is known to be a northern tick and is a rare tick in the southern part of the country. Of the 11 tick species collected, only four tick species were used in the genetic characterization, that is, Hyalomma truncatum, Hyalomma rufipes, Amblyomma hebraeum and Rhipicephalus lunulatus.  Primers OPY-3 and OPX-14 revealed genetic variation amongst the four species showing that RAPDs can be used for determining genetic diversity amongst tick population. This study forms a baseline in further studies on genetic characterization of ticks in Zimbabw