Identification of excretory secretory (ES) liquid antigen protein Fasciola gigantica with polyethilene glycol (PEG) separation

https://doi.org/10.22146/ijbiotech.27017

Yendri Junaidi(1*), Ima Malawati(2), Made Sriasih(3)

(1) Postgraduate Program of the Faculty of Animal Science, Universitas Gadjah Mada, Jalan Fauna No. 3 Bulaksumur, Yogyakarta 55281, Indonesia
(2) Postgraduate Program of the Faculty of Animal Science, Universitas Gadjah Mada, Jalan Fauna No. 3 Bulaksumur, Yogyakarta 55281, Indonesia
(3) Laboratory of Microbiology and Biotechnology, Faculty of Animal Science, Mataram University, Jalan Majapahit 62, Mataram, Nusa Tenggara Barat 83125, Indonesia
(*) Corresponding Author

Abstract


Fasciola gigantica diagnosis usually performed by detection of worm eggs presence in the feces,but this conventional method has many disadvantages. Early diagnosis (early detection) cannot be performed in conventional methods because the worms in the host's body began to lay eggs at the age of 8–12 weeks of patency. The current detection method that is based on antibody­antigen reactions using excreted/secreted (ES) liquid by adult F. gigantica, is believed to be used for the early detection of fasciolosis. This study aimed to characterize the antigenic components of F. gigantica extretory/secretory products that could be used as a vaccine candidate development for early fasciolosis diagnostics. ES products were separated by PEG4000 at various concentrations (8%, 16%, 24%), then precipitates (pellets) obtained were dialyzed and characterized using SDS­PAGE and Western blotting. Results from SDSPAGE showed that there were 18 proteins bands with 7–70 kDa molecular weights. Western blotting on pellets derived from PEG separation at various concentrations affirmed that the proteins of 50, 25 and 20 kDa were antigenic at 8% PEG concentration, the 25 kDa and 50 kDa were antigenic at 16% PEG concentrations and the 25 kDa was antigenic at 25% PEG concentration


Keywords


antigenic; diagnostic; ES liquid; Fasciola gigantica; fasciolosis; PEG4000

Full Text:

PDF


References

Abidin, Z. (2002) Laporan Tahunan Dinas Peternakan Kabupaten Gowa. Dinas Peternakan Kabupaten Gowa. Jawa Barat.

Anonymous. (2011) Pengembangan metode ELISA utuk mendeteksi keberadaan koproantigen Fasciola gigantica pada ruminansia: model uji diagnostik untuk human fasciolosis. Bogor: Sekolah Pasca Sarjana IPB.

Astiti, L.G.S., Estuningsih, E., Hartiningrum, D.B., Muliyati, K., and Ichwan (2011) Pemberantasan dan pengendalian penularan fasciolosis pada sapi bali di Pulau Lombok. NTB: Balai Pengkajian Teknologi Pertanian Nusa Teggara Barat.

Chwohury, N., and Tada I. (1994) Helminthology. New Delhi: Springer-Verlag-Norasa Publising House.

Dewi, N.C., Nurhidayati, T., Purwati, K.I. (2012) Pengaruh penambahan PEG (Polyethylen Glicol) terhadap profil protein tembakau (Nicotiana tabacum L. var Prancak 95) pada media in vitro. Surabaya: FMIPA Institut Teknologi Sepuluh November.

El-Rahimy, M. (2012) Evaluation of some Fasciola gigantica antigens as vaccines against fasciolosis in goats. Giza: Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary Medicine. Cairo University.

Estuningsih, S.E., Spithill, T., Raadsma, H., Law, R., Adiwinata, G., Meusen, L., and Piedrafitall, D. 2009. Development and application of a fecal antigen diagnostic sandwich ELISA for estimating prevalence of Fasciola gigantica in cattle in Central Java, Indonesia. J. Parasitol., 95(2), 450–455.

Garfin, D.E. (2013) Gel electrophoresis of proteins. In: Davey J., and Lord, M. (Eds) Essential cell biology vol. 1, chapter 7: cell structure, a practical approach. Oxford: Oxford University Press.

Farghaly, A.M., Nada, S.M., Emam, W.A., Matter, M.A., Mohamed, S.M.A., Sharaf, E.M., and El-Gamal, R.A. 2009. Role of Fast-ELISA and Western Blot in diagnosis of human fasciolosis using crude adult worm and excretory/secretory Fasciola antigen. Int. J. Vet. Sci. Med., 1(2), 55–65.

Guyton, A.C., and Hall, J.E. (2007) Fisiologi kedokteran edisi 11 (Translation of the Textbook of Medical Physology 11th Edition). Jakarta: EGC.

Harris, M.J. (Ed.) (1992) Poly(ethylene glycol) chemistry: biotechnical and biomedical applications. New York: Plenum Press.

Kim, K., Yang, H.J., and Chung, Y.B. 2003. Usefulness of 8 kDa protein of Fasciola hepatica in diagnosis of fasciolosis. The Korean J. Parasitol., 41(2), 121–123.

Kumar, N, Ghosh, S., and Gupta, S.C. 2008. Early detection of Fasciola gigantica infections in buffaloes by enzim linked immunosorbent assay. Parasitol. Res., 103, 141–150.

Kusnoto, Subekti, S., Sudiana, I.K., and Soedarno. 2011. Karakterisasi dan isolasi protein spesifik dari material Ekskretori-Sekretori (ES) Toxocara cati untuk pengembangan diaknostik toxocariasis dengan teknik ELISA. JBP, 13(1), 56–65.

Lastuti, N.D.R., Pasila, A.R., and Nurhayati, T. 2008. Identifikasi profil protein eksresi sekresi cacing Haemonchus contortus dewasa dengan SDS-PAGE. Veterinaria Medika, 1(1), 39–42.

Laula, P.D., Koesdarto, S., and Atik, M,G. (2007) Karakterisasi protein ekskretori sekretori larva stadium kedua dorman terhadap antibodi anti-larva stadium kedua Toxocara canis dengan teknik Western blot. Thesis. Faculty of Veterinary Medicine Airlangga University. Surabaya.

Marcodo, R. 1989. Detection of immunocreative peptides of F. gigantica with sera from infected subjects by enzyme immunoelectrotransfer. Bol. Chil. Parasitol., 44(3–4), 86–88.

Mas-Coma, S., Valero, M.A., and Bargues, M.D. 2009. Fasciola, lymnaeids, and human fasciolosis with a global overview on disease transmission epidemiology, evolutionary genetics, molecular epidemiology and control. Adv. Parasitol., 69, 41–146.

Meshgi, B., Eslami, A., and Hemmatzadeh, F. 2008. Determination of somatic and excretory-secretory antigens of Fasciola hepatica and Fasciola gigantica using SDS-PAGE. Iranian J. Vet. Res., 9(1), 77–80.

Mitchell, G.B.B. (2007) Liver fluke. In: Aitken, I.D. (Ed.). Disease of sheep 4th ed. London: Blackwell.

Mohamed, M.M., Al-Sherbiny, M.M., Sharaf, A.A., and Elmamlouk, T.H. 2004. Immunological identification of Fasciola hepatica antigens containing major human T-cell and B-cell epitopes. J. Egypt Soc. Parasitol., 34(3), 751–766.

Neverrete, P. A., Yadav, S.C., and Raina, O.K. 1993. Vaccination of buffaloes with Fasciola gigantica recombinant fatty acid binding protein. Parasitol. Res., 97, 129–135.

Priago, M.V., Valero, M.A., Panova, M., and Mas-Coma, S. 2006. Phenotypic comparison of allopatric populations of Fasciola hepatica and Fasciola gigantica from European and African bovines using a computer image analysis system (CIAS). Parasitol. Res., 99., 368–378.

Rivera, M.C.A., Santiago, N., and Hillyer, G.V. 1988. Evaluation of immunodiagnostic antigen in the ES production of Fasciola hepatica. J. Parasitol., 74(4), 646–652.

Shehab, A.Y., Hassan, E.M., Abou, B.L.M., Omar, E.A., Helmy, M.H., El-Morshedy, H.N., and Farag, H.F. 1999. Detection of circulating e/s antigen in the sera of patients with fascioliasis by ELISA: a tool of serodiagnosis and assessment of cure. Trop. Med. Int. Health, 4, 686–690.

Spithill, F.W. (1999) Fasciola gigantica: epidemiology, control, immunology and molecular biology. In: Fasciolosis. Wallingfort: CAB International Publishing.

Sriasih, M., Depamede, S.N., and Ali, M. (2013) Karakterisasi protein antigenik cairan eksretori sekretori cacing F. gigantica isolat lokal dengan teknik Western blotting. Research report. Mataram: Mataram University.

Suryana, A. (2009). Dukungan teknologi penyediaan produk pangan peternakan bermutu, aman dan halal. Jakarta: Agricultural Research and Development Agency.

WHO (Wold Health Organization). (2009) Position paper on food-borne Trematode infections and taeniasis/cysticercisis. Vientiane. Lao People’s Democratic Republic.

Wibowo, M.S. (2010) Elektriforesis. Bandung: School of Pharmacy ITB.

Yuliana. (2010) Pengaruh invigorasi menggunakan polyethylene glycol (PEG) 6000 terhadap viabilitas benih tembakau (Nicotiana tabacum). Thesis. Faculty of Science and Technology Islamic State University Maulana Malik Ibrahim. Malang.

Xi, J., Wang, X., Li, S., Zhou, X., Yue, L., Fan, J., and Hao, D. (2006) Polyethylene glycol fractionation improved detection of low-abundant proteins by two-dimensional electrophoresis analysis of plant proteome. Phytochemistry, 67(21), 2341–2348.

Zhang, W., Peigne, F., Moreau, E., Chauvin, A., Huang, W. 2005. Comparison of modulation of sheep, mouse, and buffalo lympochocyte responses by Fasciola hepatica and Fasciola gigantica excretory-secretory product. Parasitol. Res., 95(5), 333–338.



DOI: https://doi.org/10.22146/ijbiotech.27017

Article Metrics

Abstract views : 2415 | views : 2149

Refbacks

  • There are currently no refbacks.


Copyright (c) 2016 The Author(s)

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.