Articles

In vitro Effects of Triclabendazole (TCBZ) on the Excretory-Secretory Products (ESP) of Fasciola spp Parasites

Abstract

Fascioliasis is an endemic disease in Iran and triclabendazole (TCBZ) is using for treatment of domestic animals and infected people. Excretory-secretory products (ESP) play an important role in the host biochemical defense by means of activities of detoxifying and antioxidant glutathione S-transferase (GST) and superoxide dismutase (SOD) enzymes respectively. Therefore, the aim of this comparative study was to evaluate fasciola protection against TCBZ drug by detection of enzymatic activities, GST and SOD, in TCBZ treated Fasciola hepatica / Fasciola gigantica and control ESP samples. F. gigantic and F. hepatica helminthes were collected and cultured within buffer media (TCBZ treated and untreated or control) for 4 h at 37 °C. Three TCBZ treated and 1 control ESP samples for each species were collected, centrifuged and supernatants were stored at -20°C. ESP samples protein concentrations were measured by Bradford method. SOD and GST enzymes activities of ESP samples were estimated photometrically. To determine the statistically significant difference between ESP of treated and control samples, t-test was conducted. ESP protein bands were detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Protein concentrations in treated F. hepatica and F. gigantica ESP samples were estimated 204.88, 428, 130.4 and 288.2, 488.2, 308.2 µg/ml respectively. Protein concentrations in control samples were estimated 488.18 and 124.8 ug/ml respectively. SOD enzyme specific activities level in treated F.hepatica and F. gigantica ESP samples were determined 0.14, 0.31, 3.96 and 11.11, 13.54, 19.95 U/mg/protein respectively. SOD activities level in control samples were detected 70.69 and 10.92 U/mg/protein. GST specific activities level in treated F.hepatica and F. gigantica ESP samples were calculated 25.3, 85.5, 37.3 and 1823, 1314.3, 1320.8 U/mg respectively. GST activities levels in control samples were detected 98.6 and 1083.9 U/mg/protein respectively. Statistical analysis reveal the significant different between proteins concentrations, GST and SOD enzyme specific activities of TCBZ treated ESP samples of F. gigantic in comparison to the control samples (P0.05). There is no difference between SDS-PAGE results of treated and control samples. Based on the results of the present work, significant increase of enzymatic activities of GST and SOD in TCBZ treated F. gigantica ESP, it seems, the protection of this species against drug is higher than F. hepatica.

World Health Organization (WHO). Neglected tropical diseases. Fascioliasis. [Internet] 2012 [cited 2012 Feb 15]; Available from: http://www.who.int/neglected_diseases/ diseases/fascioliasis/en/

Knobloch J, Delgado E, Alvarez A, Reymann U, Bialek R. Human fascioliasis in Cajamarca/Peru. I. Diagnostic methods and treatment with praziquantel. Trop Med Parasitol 1985;36(2):88-90.

Massoud J. Fascioliasis outbreak of man and drug test (triclabendazole) in Caspian littoral, northern part of Iran. Bul Soc Fr Parasitol 1990;8:438.

Savioli L, Chitsulo L, Montresor A. New opportunities for the control of fascioliasis. Bull World Health Organ 1999;77(4):300.

World Health Organization (WHO). Action Against Worms. [Internet] 2007 Dec [cited 2012 Feb 15]; Available from: http://www.who.int/neglected_diseases/ preventive_chemotherapy/Newsletter10.pdf

Lee CG, Zimmerman GL, Mulrooney DM. Isoelectric focusing of soluble proteins from Fasciola hepatica L, 1758 and Fascioloides magna B, 1875. Am J Vet Res 1992;53(2):246-50.

Jefferies JR, Campbell AM, van Rossum AJ, Barrett J, Brophy PM. Proteomic analysis of Fasciola hepatica excretory-secretory products. Proteomics 2001;1(9):1128-32.

Abu-Amero KK, Morales J, Mohamed GH, Osman MN, Bosley TM. Glutathione S-transferase M1 and T1 polymorphisms in Arab glaucoma patients. Mol Vis 2008;14:425-30.

Sies H. Oxidative stress: oxidants and antioxidants. Exp Physiol 1997;82(2):291-5.

Chemale G, Perally S, LaCourse EJ, Prescott MC, Jones LM, Ward D, Meaney M, Hoey E, Brennan GP, Fairweather I, Trudgett A, Brophy PM. Comparative proteomic analysis of triclabendazole response in the liver fluke Fasciolahepatica. J Proteome Res 2010;9(10):4940-51.

Toner E, Brennan GP, McConvery F, Meaney M, Fairweather I. A transmission electron microscope study on the route of entry of triclabendazole into the liver fluke, Fasciola hepatica. Parasitology 2010;137(5):855-70.

Maizels RM, Blaxter ML, Robertson BD, Selkirk ME. Parasite Antigens, Parasite Genes. A Laboratory Manual for Molecular Parasitology. Cambridge University Press, 1991.

McCord JM, Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem 1969;244(22):6049-55.

Randox Life Sciences Ltd. Superoxide dismutase assay (ransod). [Internet] 2012 [cited 2012 Feb 15]; Available from: http://www.randoxonlinestore.com/RANSOD- %28Superoxide-Dismutase%29-p-7904

Minnesota State University Moorhead. Enzyme Assay for Glutathione S- Transferase Protocol. [Internet] 2007 Jan [cited 2012 Feb 15]; Available from: http://web.mnstate. edu/provost/gst%20assay%20protocol.pdf

Morphew RM, Wright HA, LaCourse EJ, Woods DJ, Brophy PM. Comparative proteomics of excretorysecretory proteins released by the liver fluke Fasciola hepatica in sheep host bile and during in vitro culture ex host. Mol Cell Proteomics 2007;6(6):963-72.

Meshgi B, Eslami A, Hemmatzadeh F. Determination of somatic and excretory-secretory antigens of Fasciola hepatica and Fasciola gigantica using SDS-PAGE. IJVR 2008;9(1):77-80.

Gourbal BE, Guillou F, Mitta G, Sibille P, Thèron A, Pointier JP, Coustau C. Excretory-secretory products of larval Fasciola hepatica investigated using a twodimensional proteomic approach. Mol Biochem Parasitol 2008;161(1):63-6.

Shehab AY, Ebeid SM, El-Samak MY, Hussein NM. Detoxifying and anti-oxidant enzymes of Fasciola gigantica worms under triclabendazole sulphoxide (TCBZSX): an in vitro study. J Egypt Soc Parasitol 2009;39(1):73-83.

Cervi L, Rossi G, Masih DT. Potential role for excretorysecretory forms of glutathione-S-transferase (GST) in Fasciola hepatica. Parasitology 1999;119 ( Pt 6):627-33.

Ganga G, Varshney JP, Patra RC. Activity of antioxidant enzymes in excretory-secretory fluid and somatic extracts of Fasciola gigantica. J Vet Parasitol 2007;21(1):51-2.

Toner E, Brennan GP, Hanna RE, Edgar HW, FairweatherI. Fasciola hepatica: time-dependent disruption of spermatogenesis following in vivo treatment with triclabendazole. Parasitol Res 2011;109(4):1035-43.

Files
IssueVol 50, No 3 (2012) QRcode
SectionArticles
Keywords
Fasciola Excretory-secretory product Triclabendazole

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Farahnak A, Golmohamdi T, Eshraghian M. In vitro Effects of Triclabendazole (TCBZ) on the Excretory-Secretory Products (ESP) of Fasciola spp Parasites. Acta Med Iran. 1;50(3):164-168.