Abstract. A method for gravimetric determination of tannins based on binding with insoluble polyvinylpyrrolidone (PVP) is presented. The gravimetric method gives the absolute amount of tannins and avoids problems of standards associated with spectrophotometric methods. The method was applied to nine browse and tree leaves. The values obtained correlate significantly with tannins determined spectrophotometrically, protein precipitation capacities and protein precipitable phenolics. This method together with other tannin assays will be useful in nutritional studies. The present study also demonstrates the different behaviour of tannic acids from different commercial sources towards PVP suggesting the presence of different moieties in tannic acids from different commercial sources and even among batches from the same source thereby affecting the results obtained using the spectrophotometric methods. Use of well-defined tannic acid as a standard in spectrophotometric methods is suggested which will allow meaningful comparison of values obtained from different laboratories.

A method for the selective depletion of transferrin from bovine serum is presented. Bloodstream forms of Trypanosoma brucei cannot grow in medium containing transferrin-deficient serum, whereas reconstitution with transferrin restores normal growth. We conclude that transferrin is an essential growth factor for the mammalian stage of the parasite.

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Intraperitoneal injection of Trypanosoma brucei AnTat 1.1 into mice of the C3H.He, BALB/c or C57BL/6 strains resulted in impaired immune responses from day 3 onwards, as measured by the reduction in DNA synthesis in spleen cell populations stimulated with concanavalin A (Con-A) in vitro. Adherent cells from the peritoneum (PC) or from the spleen of infected mice, consisting predominantly of macrophages, caused a 60-80% reduction of the Con-A response in spleen cells from syngeneic recipients 3-4 days after transfer in vivo. Adherent PC from irradiated or athymic mice were equally suppressive. Spleen cells from infected mice reduced the proliferative response of spleen cells from uninfected mice upon co-cultivation in vitro. This dominant suppressive effect was abolished after the selective removal of macrophages from the spleen cell population by treatment with L-leucine methylester. Moreover, the macrophage-depleted spleen cells from infected mice responded normally to Con-A provided they were supplemented with splenic adherent cells from naive mice as a source of accessory cells. Both the cell transfer and co-cultivation experiments suggest that infection with African trypanosomes changes the properties of macrophages to a state which allows them actively to suppress immune responses.