IJSTR

International Journal of Scientific & Technology Research

Home About Us Scope Editorial Board Blog/Latest News Contact Us
0.2
2019CiteScore
 
10th percentile
Powered by  Scopus
Scopus coverage:
Nov 2018 to May 2020

CALL FOR PAPERS
AUTHORS
DOWNLOADS
CONTACT

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]

 

AUTHOR(S)

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

 

KEYWORDS

Midgut, property, trypanolysin, trypanosomes, tsetse.

 

ABSTRACT

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.

 

REFERENCES

[1] Abubakar et al., 1995. "Properties of bloodmeal-induced midgut lectin from the tsetse fly, Glossina morsitans. Parasitology Research".

[2] Abubakar, L.U.; Bullmo, W.D.; Muloo, F.J. and Osir, E.O. (2006). "Molecular characterization of a tsetse fly midgut proteolytic lectin that mediates differentiation of African trypanosomes". Insect Biochem.Mol.Biol.36, 344-352.

[3] Acosta-Scerrano, A.; Vassella, E., Linger, M.; Renggli, C.K.; Brun, R.; Roditi, I. and Englund, P.T. (2001). "The surface coat of procyclic T.brucei; programmed expression and proteolytic cleavage of procyclin in the tsetse fly", Proc.Natl.Sci.USA 98, 1512-1518.

[4] Aksoy, S. 2000. "Tsetse - a haven for microorganisms". Parasitol. Today 16: 114-118.

[5] Brun & Jenni (1977). "A new semi-defined medium from Trypanosoma brucei spp". Acta Trop. (Basel).

[6] Brun & Schonenberger, (1970). "Cultivation and in vitro clonig of procyclic culture forms of trypanosome brucei in a semi-defined medium". Acta Trop. (Basel).

[7] Cheesememan, M.T. and Gooding, R.H (1985). Proteolyic enzymes from tsetse flies Glossina palpalis (Diptera:Glossinidae). Insect. Biochem.

[8] Duke, H.L (1936). "On the prophylactic action "Bayer 205" against the trypanosomes of man." Lancet.

[9] Gibson, W. and Bailey, M. (2003). "The development of T.brucei within the tsetse fly midgut observed using green fluorescent trypanosomes". KinetoplastidBiol.Dis. 2, 1-13.

[10] Goding, R.H. (1974a). "Digestive process of Hematophagous insects. XI. Partial purification and some properties of six proteolytin enzymes from the tsetse fly Glossina morsitans morsitans West wood (Dipetar:Glossinidae)".

[11] Hao, Z.; Kasumba, I.; Lehane, M.J.; Gibson, W.C.; Kwon, J. and Aksoy, S. (2001). "Tsetse immune responses and trypanosome transmission: implications for the development of tsetse based strategies to reduce trypanosomiasis". Proceedings of the National Academy of sciences of the USA, 98, 12648-12653.

[12] Imbuga, M.O., Osir, E.O. and Labongo, V.L., Daryi, N. and Otieno, L.H (1992a). "Studies on tsetse midgut factors that induce differentiation of bloodmeal of Trypanosomes brucei in vitro Parasitol".

[13] Imbuga, M.O., Osir, E.O. and Labongo, V.L., (1992b). "Inhibitory effect of Trypanosome brucei brucei on Glossina morsitans morsitans midgut trypanosin in vitro Parasitol".

[14] Maudlin, I. (1882). "Inheritance of susceptibility to trypanosomes infection in Glossina morsitans". Ann. Trop. Med. Parasitol.

[15] Maudlin, I. (1991). "Transmission of African trypanosomias among tsetse immune system, symbionts and parasites Adv.Disease Vector Res".

[16] Moolo, S.K., and Kutuza, S.B and Desai, J. (1988b). "Infection rates in sterile males of Glossina morsitans, Glossina palpalis and fuscs groups of Glosina austeni and Glossina tachinoides for pathogenic trypanosomes species from East and West Africa trypanosome vivax stockes, Parasitology".

[17] Ochieng Nyambega B.C. "The mechanism of Trypanolysin mediated lysis of T. b. brucei" (Master of Science, 2002, Nairobi, Kenya-JKUAT).

[18] Osir, E.O. Abubakar, L. and Imbuga, M.O. (1995). "Purification and characterisation of a midgut lectin-trypsin complex from the tsetse fly, Glossina longipennis Parasitol Res".

[19] Osir, E.O. Abubakar, M.O and Onyango.P. (1993). "Inhibition of Glossina morsitans morsitans midgut trypsin activity by D-glucosamine. Parasitol".

[20] Otieno,L.H., Darji, N.,Onyango,P.and Mpanga,I.(1983). "Some observations on factors associated with the development of Trypanosoma brucei brucei infection in Glossina morsitans morsitans". Acta Trop.,

[21] Stiles, J.K.,Ingram, G.A., Walbanks, K.R., Molyneux, D.H.,Maudlin, I., and Welburn, S.(1990). "Identification of midgut trypanolysin and trypanoagglutinin in Glossina palpalis sspp.(Diptera:Glossinidae) Parasitol".

[22] Stiles,J.K., Walbanks, K.R., and Molyneux, D.H.(1991). "The use of casein subtrate gels for determining trypsin-like activity in the kidgut of Glossina palpalis sspp".

[23] Van den Abbelle, V.J., Declair, W (1992). "Studies of vectorial capacity of Glossina sp.related to its digestive physiology and rearing conditions.tsetse control diagnosis and chemotherapy using nuclear techniques IAEA-TECDOC". International Energy Agency, Vienna.

[24] Vassella, E.; Acosta-Scerrano, A; Studer, E.; Lee, S.H.; Englund, P.T. and Roditi, I. (2001). "Multiple procyclinis forms are expressed differentially during the development of insect forms of T.brucei". J.Mol.Biol. 312, 597-607.