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IJSTR >> Volume 5 - Issue 10, November 2016 Edition

International Journal of Scientific & Technology Research  
International Journal of Scientific & Technology Research

Website: http://www.ijstr.org

ISSN 2277-8616

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

[Full Text]



Mahamat H.Abakar, Ellie O.Osir, Mabel O.Imbuga, Hector G.Morgan



Midgut, property, trypanolysin, trypanosomes, tsetse.



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 27℃ and 4℃ after 15 and 32 days, respectively. Similarly, heating the midguts trypanolysin to 60 - 80℃ led to loss of activity. On the other hand, 50℃ was found to be the optimum temperature for trypanolysin activity. The activity was also unstable by freeze-thaw at 80℃, -70℃, -20℃ and 0℃ 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.



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