The profiles of semen sialic acid and the enzyme alpha-L-fucosidase were studied in rams undergoing chronic infection by Trypanosoma congolense. Our data showed a significant surge in the level of sialic acid with parasitaemia. The pattern followed a polynomial function we had reported for erythrocyte sialic acid in mice undergoing acute infection by T. congolense. The activity of the enzyme alpha-fucosidase decreased progressively with approximately 60% decrease at the end of the 14 weeks of infection. Representative semen samples from the control and infected rams were subjected to kinetic characterization. While the uninfected semen sample showed two active pH peaks at 4.5-5.5 and at 6.8-7.2, respectively, there was an apparent shift to only a single pH optimum at 4.5-5.5 for the pathological semen. The fucosidases from both sources were optimally active at 35 degrees C albeit with contrasting activation energies (E(a)) with values 20.58 and 35 kJ/mol for the control and infected semen, respectively. Kinetic studies using methylumbelliferyl-beta-fucoside (4MU-Fuc) as substrate gave K(M) and V(max) values of 3.25 microM and 14.6 micromol. min(-1) mg(-1), respectively for the control semen. The values for the infected semen were 18.25 microM and 10.5 micromol. min(-1) mg(-1), respectively. The significance of these results is discussed as they relate to loss in reproductive capacity in trypanosomoses.
Pearl millet is widely grown as a multi-purpose cereal grain crop for feed, fodder, fuel and mulch predominantly in the semi-arid tropics. It is highly adapted to drought, representing an essential component of the food security and livelihood of many million poor farmers. DNA extraction is unusually difficult in some plants due to the presence of secondary metabolites that interfere with DNA isolation making it very laborious and time consuming. An effective genomic DNA extraction should be simple, cost effective, with good yield and high purity. The protocol used in this study involved the extraction of genomic DNA from fresh leaves using the Sodium dodecyl sulphate (SDS) method with slight modifications including absence of use of liquid nitrogen which is difficult and expensive to obtain in the developing World. Genomic DNA obtained from the ten pearl millet samples using this procedure was good.
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