Molecular characterisation of Trypanosoma evansi stocks from Indonesia

• Main Author: Sukanto, Ismu Prastyawati
• Published: University of Edinburgh 1999
• Subjects: 576.58
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.662591

The work described in this thesis used the techniques of karyotype analysis, random amplified polymorphic DNA analysis (RAPD), Riboprinting and simple sequence repeat polymerase chain reaction (SSR-PCR) to examine genetic variation in stocks of Indonesian <I>Trypanosoma evansi</I>. The 80 <I>T. evansi</I> stocks investigated were isolated from natural infections in cattle, buffaloes and horses in 10 widely separated areas, including Aceh, North Sumatra, Lampung, West, Central and East Java, Madura, South Kalimantan, North and South Sulawesi. Transverse alternating field electrophoresis (TAFE) was used to determine the polymorphisms present in the karyotype patterns of the 80 <I>T. evansi</I> stocks. Electrophoretic conditions were developed to give good resolution of chromosome bands in the size range 50 kb to 5.7 Mb. The total number of chromosome bands in the <I>T. evansi</I> genome varied from 17-27 depending on the stock. A high degree of polymorphism was detected in the karyotype patterns of <I>T. evansi</I> stocks from Indonesia by TAFE. In the 80 stocks studied, 46 different karyotype patterns were detected. Cluster analysis with a grouping level of 10% fitted the patterns into seven main karyotype groups which correlated with the locality from which the stocks had been isolated. The stability of the karyotype patterns within stocks was not related to differences between the surface variant antigenic types (VAT) in that relapse populations derived from a single infection with a cloned stock of <I>T. evansi</I> showed the identical karyotype pattern to the original infecting stock. In relapse populations collected from an uncloned <I>T. evansi</I> stock different karyotype patterns were found indicating that more than one population was present in the original isolate. Attempts to isolate VAT specific chromosomal probes were not successful.