Properties Of A Midgut Trypanolysin From The Tsetse Fly Glossina Morsitans Morsitans

The properties of a bloodmeal-induced trypanolysin from the midgut of the tsetse G. m. morsitans was studied in vitro. The semi-purified trypanolysin from twice-fed tsetse had the highest trypanolysin activity against bloodstream trypanosomes followed by those once-fed and the unfed flies. Serum found to display trypanolysin activity. The trypanolysin had no trypsin activity nor even affected by the enzyme. In addition trypanolysin was not affected by protease inhibitors such as soy bean trypsin inhibitor STI N-a-p-Tosyl-L-lysine chromethyl ketone TLCK phenylmethyl sulphonyl fluoride PMSF diisopropyl fluoro-phosphate DFP and tosylamide-2-phenylethyl chloromethyl ketone TPCK. However the activity was completely inhibited by diethyl pyrocarbonate DEPC and partially by aprotinin. The induction of trypanolysin activity by bloodmeal increased gradually reaching a peak at 72-120 h after the bloodmeal and then decreased rapidly with only 25 of the peak activity remaining after 192 h. The trypanolysin was inactivated during storage at 27268451 and 4268451 after 15 and 32 days respectively. Similarly heating the midguts trypanolysin to 60 - 80268451 led to loss of activity. On the other hand 50268451 was found to be the optimum temperature for trypanolysin activity. The activity was also unstable by freeze-thaw at 80268451 -70268451 -20268451 and 0268451 after 33 41 55 and 63 days respectively. Trypanolysin caused lyses of bloodstream-form T. b. brucei while the procyclic trypanosomes were unaffected. The highest trypanolysin activity in different tsetse species was found with Glossina longipennis followed by Glossina pallidipes Glossina morsitans centralis Glossina fuscipes fuscipes and G. m. morsitans. When the midgut homogenate was separated by anion-exchange chromatography the trypanolysin activity was recovered in the bound fraction. These results suggest that the midgut trypanolysin plays an important role in the establishment of trypanosome infections in tsetse.