Development of Dot Blot Technique for Detection of Vitellogenin as a Biomarker of Endocrine Disrupting Chemicals
Abstract
Development of a dot blot technique for detection of vitellogenin in sea bass (Lates calcarifer) for using as a biomarker of endocrine disrupting chemicals (EDCs) exposure to the environment. Dot blot technique by using a specific monoclonal antibody to vitellogenin of sea bass (MAb-sea bass VTG F3-23) can be measured vitellogenin levels at 7.8 ng. It can be used to monitor vitellogenin in natural fish. Cross-reactivity testing of MAb-sea bass VTG F3-23 has shown the reactivity to vitellogenin from Coral grouper and Nile tilapia that were induced by injection with 17β-estradiol without cross-reactivity to control fish. The assessment of vitellogenin in 6 fish species (nile tilapia, three spotted tilapia, striped snake-head, moonbeam gourami, climbing perch, and silver barbs) from industrial, residential and agriculture area reveals that, the highest induction of VTG in male fishes were found from the industrial and agriculture area respectively (33.33% and 11.11%). These data suggest that EDCs were present in aquatic environment and vitellogenin detection using dot blot technique could be validated for a biomarker of EDCs in aquatic environment. Keywords : vitellogenin, sea bass, biomarker, dot blot, endocrine disrupting chemicalsReferences
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Bastos, C.R., Mathias,L. A., Gomes-Jusi, M.M., Ferreira dos Santos, R., Pinheiro da Silva, G.C., André, M.R., Machado, R.Z., & Bürger, K.P. (2018). Evaluation of dot-blot test for serological diagnosis of bovine brucellosis. Brazilian Journal of Microbiology. (In press).
Bjerregaard, P., Hansen, P.R., Larsen, K.J., Erratico, C., Korsgaard, B., & Holbech, H. (2008). Vitellogenin as a biomarker for estrogenic effects in brown trout, Salmo trutta: laboratory and field investigations. Environmental Toxicology and Chemistry, 27, 2387–2396.
Combalbert, S., & Hernandez-Raquet, G. (2010). Occurrence, fate, and biodegradation of estrogens in sewage and manure. Applied Microbiology and Biotechnology, 86, 1671–1692.
Duong, C.N., Ra, J.S., Cho, J., Kim, S.D., Choi, H.K., Park, J-H., Kim, K.W., Inam, E., & Kim, S.D. (2010). Estrogenic chemicals and estrogenicity in river waters of South Korea and seven Asian countries. Chemosphere, 78, 286–293.
Garnayak, S.K, Mohanty, J., Rao, T.V., Sahoo, S.K., & Sahoo, P.K. (2013). Vitellogenin in Asian catfish, Clarias batrachus: Purification, partial characterization and quantification during the reproductive cycle by ELISA. Aquaculture, 392-395, 148-155.
Jackson, J., & Sutton, R. (2008). Sources of endocrine-disrupting chemicals in urban wastewater, Oakland, CA. Science of the Total Environment, 405(1-3), 153–160.
Lomax, D.P., Roubal, W.T., Moore, J.D., & Johnson, L.L. (1998). An enzyme-linked immunosorbent assay (ELISA) for measuring vitellogenin in English sole (Pleuronectes etulus): development, validation and cross-reactivity with other pleuronectids. Comparative Biochemistry and Physiology Part B, 121, 425–436
Morthorst, J.E., Mathiesen, K.K., Holbech, H., Pedersen, K.L., & Bjerregaard, P. (2017). Vitellogenin concentrations in feral Danish brown trout have decreased: An effect of improved sewage treatment in rural areas? Environmental Toxicology and Chemistry, 37, 839–845.
Nanthanawat, P. (2015). Vitellogenin : Biomarker for EDCs Exposure in Aquatic Environment. Burapha Science Journal, 20(2), 201-208. (in Thai)
Nanthanawat, P., Khongchareonporn, N., & Srisawas, S. (2016). Development of monoclonal antibody specific to plasma Vitellogenin in Asian sea bass (Lates calcarifer Bloch) Full report. (The Research grant of Burapha University through the National Research Council of Thailand, grant no. 79/2558). Chon Buri, Thailand: Burapha University. (in Thai)
Peter, L.D., Doyottea, A., Mitchelmorea, C.L., McEvoy, J., & Livingstonea, D.R. (2001). Seasonal variation and estradiol-dependent elevation of Thames estuary eel Anguilla anguilla plasma vitellogenin levels and comparisons with other United Kingdom estuaries. The Science of the Total Environment, 279, 137–150.
Prasatkaew, W., Thanomsit, C., & Nanthanawat, P. (2016). Cross-Reactivity of Monoclonal Antibody against Plasma Vitellogenin from Asian Sea Bass (Lates calcarifer) In Proceedings the 7th National Science Research Conference. (pp. 1–7). Phitsanulok: Naresuan University. (in Thai)
Ritphoe, S., Pimprong, A., Prasatkaew, W., & Nanthanawat P. (2015).Purification and Characterization of Vitellogenin from Asian Sea bass. In Proceedings the 8th Science Research Conference. (pp. 491–496). Payao : University of Payao. (in Thai)
Rupprecht, K.R., Nair, R.K., Harwick, L.C., Grote, J., Beligere, G.S., Rege, S.D., Chen, Y.Y., Lin, Z., & Fishpaugh, J.R. (2010). Development of a dot-blot assay for screening monoclonal antibodies to low-molecular-mass drugs. Analytical Biochemistry, 407, 160–164.
Scott, A.P., Sanders, M., Stentiford, G.D., Reese, R.A., & Katsiadaki, I. (2007). Evidence for estrogenic endocrine disruption in an offshore flatfish, the dab (Limanda limanda L.). Marine Environmental Research, 64,
128–148.
Yamamoto, A., Garcia, J.R.E., Kupsc, A., & Oliveira Ribeiro, C. A. (2017). Vitellogenin levels and others biomarkers show evidences of endocrine disruption in fish species from Iguaçu River – Southern Brazil. Chemosphere, 186, 88–99.
148-153.
Bastos, C.R., Mathias,L. A., Gomes-Jusi, M.M., Ferreira dos Santos, R., Pinheiro da Silva, G.C., André, M.R., Machado, R.Z., & Bürger, K.P. (2018). Evaluation of dot-blot test for serological diagnosis of bovine brucellosis. Brazilian Journal of Microbiology. (In press).
Bjerregaard, P., Hansen, P.R., Larsen, K.J., Erratico, C., Korsgaard, B., & Holbech, H. (2008). Vitellogenin as a biomarker for estrogenic effects in brown trout, Salmo trutta: laboratory and field investigations. Environmental Toxicology and Chemistry, 27, 2387–2396.
Combalbert, S., & Hernandez-Raquet, G. (2010). Occurrence, fate, and biodegradation of estrogens in sewage and manure. Applied Microbiology and Biotechnology, 86, 1671–1692.
Duong, C.N., Ra, J.S., Cho, J., Kim, S.D., Choi, H.K., Park, J-H., Kim, K.W., Inam, E., & Kim, S.D. (2010). Estrogenic chemicals and estrogenicity in river waters of South Korea and seven Asian countries. Chemosphere, 78, 286–293.
Garnayak, S.K, Mohanty, J., Rao, T.V., Sahoo, S.K., & Sahoo, P.K. (2013). Vitellogenin in Asian catfish, Clarias batrachus: Purification, partial characterization and quantification during the reproductive cycle by ELISA. Aquaculture, 392-395, 148-155.
Jackson, J., & Sutton, R. (2008). Sources of endocrine-disrupting chemicals in urban wastewater, Oakland, CA. Science of the Total Environment, 405(1-3), 153–160.
Lomax, D.P., Roubal, W.T., Moore, J.D., & Johnson, L.L. (1998). An enzyme-linked immunosorbent assay (ELISA) for measuring vitellogenin in English sole (Pleuronectes etulus): development, validation and cross-reactivity with other pleuronectids. Comparative Biochemistry and Physiology Part B, 121, 425–436
Morthorst, J.E., Mathiesen, K.K., Holbech, H., Pedersen, K.L., & Bjerregaard, P. (2017). Vitellogenin concentrations in feral Danish brown trout have decreased: An effect of improved sewage treatment in rural areas? Environmental Toxicology and Chemistry, 37, 839–845.
Nanthanawat, P. (2015). Vitellogenin : Biomarker for EDCs Exposure in Aquatic Environment. Burapha Science Journal, 20(2), 201-208. (in Thai)
Nanthanawat, P., Khongchareonporn, N., & Srisawas, S. (2016). Development of monoclonal antibody specific to plasma Vitellogenin in Asian sea bass (Lates calcarifer Bloch) Full report. (The Research grant of Burapha University through the National Research Council of Thailand, grant no. 79/2558). Chon Buri, Thailand: Burapha University. (in Thai)
Peter, L.D., Doyottea, A., Mitchelmorea, C.L., McEvoy, J., & Livingstonea, D.R. (2001). Seasonal variation and estradiol-dependent elevation of Thames estuary eel Anguilla anguilla plasma vitellogenin levels and comparisons with other United Kingdom estuaries. The Science of the Total Environment, 279, 137–150.
Prasatkaew, W., Thanomsit, C., & Nanthanawat, P. (2016). Cross-Reactivity of Monoclonal Antibody against Plasma Vitellogenin from Asian Sea Bass (Lates calcarifer) In Proceedings the 7th National Science Research Conference. (pp. 1–7). Phitsanulok: Naresuan University. (in Thai)
Ritphoe, S., Pimprong, A., Prasatkaew, W., & Nanthanawat P. (2015).Purification and Characterization of Vitellogenin from Asian Sea bass. In Proceedings the 8th Science Research Conference. (pp. 491–496). Payao : University of Payao. (in Thai)
Rupprecht, K.R., Nair, R.K., Harwick, L.C., Grote, J., Beligere, G.S., Rege, S.D., Chen, Y.Y., Lin, Z., & Fishpaugh, J.R. (2010). Development of a dot-blot assay for screening monoclonal antibodies to low-molecular-mass drugs. Analytical Biochemistry, 407, 160–164.
Scott, A.P., Sanders, M., Stentiford, G.D., Reese, R.A., & Katsiadaki, I. (2007). Evidence for estrogenic endocrine disruption in an offshore flatfish, the dab (Limanda limanda L.). Marine Environmental Research, 64,
128–148.
Yamamoto, A., Garcia, J.R.E., Kupsc, A., & Oliveira Ribeiro, C. A. (2017). Vitellogenin levels and others biomarkers show evidences of endocrine disruption in fish species from Iguaçu River – Southern Brazil. Chemosphere, 186, 88–99.
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2018-10-08
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บทความวิจัยจากการประชุมวิชาการระดับชาติ"วิทยาศาสตร์วิจัย"ครั้งที่ 10
