ENZYME ELECTROPHORESIS OF SOUTH AFRICAN SCWSTOSOMA MATTHEEI AND

KRUGER, F. J., 1987. Enzyme electrophoresis of South African Schistosoma mattheei and S. haematobJum. Onderstepoort Journal ofVeterinary Research, 54, 93-96 (1987) . ~ a. pilot proj~ of a study undertaken to determine the influence of S. mattheei X S. haematobium hybridiza~on on vanous South AfricanS. mattheei populations by means of biochemical-taxonomic methods a co! Dparative electrophoretic study of laboratory-maintained S. mattheei and S. haematobium was performed ! ISmg II. enzymes represe~ting 16 gene loci. Eleven loci were found to be monomorphic, while 5 differed m~pecifically. Computation of the results revealed that South AfricanS. mattheei and S. haematobium are fmly closely related when compared with other Schistosoma spp. groups. INTRooucnoN . As enzymes are transcribed from the DNA contained m ~e nucleus of an organism, the electrophoretic properties of an enzyme provide a method by which to ~haracterize the gene cOding for the enzyme. By studymg a ~ge of enzymes in 2 animal spec1es an empirical expresston can be obtained as to the relationship between the species. Kruger, Schutte, Visser & Evans (1986) and Kruger, Hamilton-Attwell & Schutte (1986) observed that populations of the bovine blood fluke Schistosoma mattheei which occur sympatric with the human l'arasite S. hae"}'ltobium, have more phenotypic qualities m common With ~. haemato~ium than allopatric populations of ~· mattheel. They attributed the presence of these qualities to S. mattheei x S. haematobium hybrids from human origin that have reinfected cattle and introduced S. haematobium genes into the genepool of the sympatric ~· Tnflttheei.populations . Th~ most widely used technique m b10chemtcal systematics IS electrophoresis (Ferguson, 1980). It was therefore selected as the technique for further study of these populations. The electrophoretic properties of the enzymes of S. haematobium from Africa and the Middle East differ from region to region (Wright & Ross, 1983). Before the ultimate objective, viz. the measurement of the effect of hybridization on the gene pool of S. mattheei, could be achieved, a baseline comparison of South African S. mattheei and S. haematobium was essential. In this paper the genotypic and phylogenetic relationships between S. mattheei and the local strain of S. haematobium were calculated from the degree of correspondence between a selected range of enzymes. MATERIAL AND METHODS . S. mattheei used in this study was isolated from cattle m the Eastern Transvaal and maintained in Praomys (Mastomys) coucha. S. haematobium was obtained from man, also from the Eastern Transvaal and was maintained in Saccostomus campestris. Bulinus (Physopsis) globosus was used as intermediate host snail for both species. Adult male schistosomes required for electrophoresis was obtained by employing the perfusion technique of Jackson, Dettman & Higgens-Opitz (1982). Electrophoresis was performed on 220 mm X 140 mm X 2 mm, 12,5 % horizontal starch gels. For each enzyme, the electrophoretic patterns of at least 10 adult males were studied individually. Single worms were macerated in 10 J.d of distilled water and the homogenate Received 10 November 1986---Editor 93 absorbed into 5 mm strips of crochet cotton which was inserted into cuts made across the width of the gel. Four ' enzymes were studied with the homogenate obtained from a single worm. Fourteen enzyme systems were studied initially. Eleven of these were selected for further study, since they developed relatively strong and clear electrophoretic patterns. They are: acid phosphatase (ACP), aldolase (ALD), glucose-6-phosphate dehydrogenase (G6PD), glutamate oxalo acetate transaminase (GOT), hexokinase (HK), lactate dehydrogenase (LDH), leucylglycyl glycine aminopeptidase (LGG), malate dehydrogenase (MDH), 1-naphthyl acetate esterase (EST), octanol dehydrogenase (ODH) and phosphoglucomutase (PGM). The electrode and gel buffer systems used and the references containing the enzyme staining solutions are given in Table 1. Except for LGG, for which a 50 me agar overlay was used, all stains were made up to 100 me. Electrophoresis was conducted at 16 V per em gel at 8 oc for approximaely 3 h. Gels were incubated at 37 oc in the dark until the banding patterns became visible, after which they were photographed. Nei's (1972) indices of.genetic distance (D) and normalized genetic identity (I) were calculted using Green's (1979) BASIC programme.