Toxicity Level of 2, 4-D Dimethylammonium in Nile tilapia and Acetylcholinesterase (AChE) Expression (Biomarker) to Identify Exposure in Sub-lethal Concentration
Abstract
The objectives of this study were to assess toxicity of the herbicide 2, 4-D dimethyl ammonium in Nile tilapia and to examine acetylcholinesterase expression (bio-indicators) for indicating exposure in sub-lethal concentrations in laboratory condition. The results can be applied as fundamental knowledge for determining herbicide 2, 4-D dimethyl ammonium in the aquatic environment. The toxicity of this compound in Nile tilapia was estimated from the cumulative mortality which was found to increase with time and concentration of exposure. After studying the lethal toxicity, the toxicity in sub-lethal level was also evaluated which was at a concentration of 5 µl/L. Nile tilapia which exposed to herbicide in sub-lethal concentration had both behavioral and morphological changes differed from the control group depending on exposure time. Acetylcholinesterase found in tissues of brain, gill and muscle have a molecular weight of 71 kDa. The expression of acetylcholinesterase in all tissues was reduced with prolonged exposure. In changed gill tissues, we found hyperplasia, epithelial lifting, partial fusion of lamellae, edema and lamellae disorganization and blood congestion. In muscle tissue, it was found dilation of muscle fiber, spiting of muscle, and blood congestion. These tissue alterations increased with an increasing in exposure time. Based on all information we found, tilapia can be used as bio-indicator for herbicide 2, 4-D dimethyl ammonium contamination in water resources. Moreover, acetylcholinesterase expression can also be used as a biomarker of exposure. Keywords : herbicide ; 2, 4-D dimethylammonium ; biomarker ; acetylcholinesteraseReferences
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Samanta, P. Rituparna, D.A.S., Sandipan, P.A.L., Aloke kumar, M., Senapati, T., Kole, D & Ghosh, A.R. (2018).
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Sarikaya, R. & Selvi, M. (2005). Investigation of acute toxicity of (2, 4-dichlorophenoxy) acetic acid (2, 4-D) herbicide
on larvae and adult Nile tilapia (Oreochromis niloticus L.). Environmental Toxicology and Pharmacology, 20, 264-268.
Sarikaya, R., & Yilmaz, M. (2003). Investigation of acute toxicity and the effect of 2, 4-D (2,4
dichlorophenoxyacetic acid) herbicide on the behavior of the common carp (Cyprinus carpio L., 1758 ; Pisces, Cyprinidae). Chemosphere, 52, 195-201.
Thanomsit, C., Maprajuab, A., Saowakoon, A., Prasartkaew, W., Ocharoen, Y., Wattanakornsiri, A., Nanuam, J., &
Nanthanawat, P. (2018). Acetylcholinesterase (AChE): Potential Biomarker for evaluating Pesticide exposure on egg and tissue of golden apple snail (Pomecea canliculata) from Huai-Saneng Resevior, Surin Province, Thailand. Thai Journal of Agricultural science, 51(3), 104-117.
Thanomsit, C., Saowakoon, S., Wattanakornsiri, A., Nanuam, J., Prasatkaew, W., Nanthanawate, P., Monkolvai, P.
& Chalorcharoenying, W. (2020a). Glyphosate (Roundup): Fate, Toxicity Assessment and Adverse Effect on Aquatic Environment. Naresuan University Journal: Science and Technology, 28(1), 65-81.
Thanomsit, C., Nuankaew, C., Prasatkaew, W., Wattanakornsiri, A., Nanuam, J., Nanthanawat, P. & Kiatprasert, P.
(2020b). Adverse effects of chlorpyrifos and cypermethrin mixture on physiological alterations and
cholinesterase on Nile tilapia (Oreochromis niloticus). Thai journal of Agricultural science, 53(3), 134-148.
Thanomsit, C., Saowakoon, K., Wattanakornsiri, A., Khongchareonporn, N., Nanuam, J., Prasatkaew, W., &
Nanthanawat, P. (2020c) Acetylcholinesterase (AChE) Polyclonal Antibody from Hybrid catfish (C. macrocephalus × C. gariepinus): Specification, Sensitivity and Cross Reactivity. Biochemistry and Physiology Part C: Toxicology & Pharmacology, 108837.
Thanomsit, C., Wattanakornsiiri, A. & Nanthanawat, P. (2016a). Effect of glyphosate on fish behavior and
histological alteration of gills in Asian Sea bass (Lates calcarifer). Burapha Science Journal, 21 (2), 204-215.
Thanomsit, C., Wattanakornsiiri, A. & Nanthanawat, P. (2016b). Toxicity evaluation of glyphosate on mortality rate and histological alteration in Black Rice crab. Burapha Science Journal, 21(3), 151-165.
U.S. EPA. (United states Environment Protection Agency). 2005. Reregistration Eligibility Decision for 2, 4-D. USA.
321 p.
Vigario, A, F. & Saboia-Morais, S.M.T. (2014). Effect of the 2, 4-D herbicide on gills epithelia and liver of the fish
Poecilia vivipara. Brazilian Journal of Veterinary Research, 34(6), 523-528.
indicator of peroxisomal proliferation in the mummichog, Fundulus heteroclitus. Marine Environmental Research, 50, 361-365.
Ateeq, B., Farah, M. A. & Ahmad, W. (2006). Evidence of apoptotic effects of 2, 4-D and butachlor on walking
catfish, Clarias batrachus, by transmission electron microscopy and DNA degradation studies. Lift Sciences, 78, 977-986.
Barbieri, E. (2008). Effect of 2, 4-D herbicide (2, 4-dichloro phenoxyacetic acid) on oxygen consumption and
ammonium excretion of juveniles of Geophagus brailiensis (Quoy & Gaimard, 1824) (Osteichthyes, Cichlidae). Ecotoxicology, 1-6.
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)
Braz-Mota, S., Sadaus=Henrique, H., Duarte, R. M., Val, A. L. & Almeida-Val, V. M. F. (2015). Roundup exposure
promotes gills and liver impairments, DNA damage and inhibition of brain cholinergic activity in the Amazon teleost fish Colossoma macroponum. Chemosphere. 135, 53-60.
Cessna, A. J., Donald, D.B., Bailey, J., Waiser. M., Headley, J. V. (2006). Persistence of the sulfonylurea
herbicides thifensulfuron-methyl, ethametsulfuron-methyl and metsulfuron-methyl infarm dugouts. Journal of Environmental Quality, 35, 2395–2401.
Castro Marcato, A.C., Pereira de souza, C. & Fontanetti, C.S. (2017). Herbicide 2, 4-D: A review of toxicity
on Non-target organisms. Water Air Soil and Pollution, 228(120), 1-12.
Cattaneo, R., Loro, V.L., Spanevello, R., Silverira, F. A., Luz, L. & Miron, D. S. (2008). Metabolic and histological
parameters of silver catfish (Rhamdia quelen) exposed to commercial formulation 0f 2, 4-dichlorophenoxiacetic acid (2,4-D) herbicide. Pesticide Biochemistry and Physiology, 92, 133-137.
Heggsteom, M. J. (2009). The sublethal effects of 2, 4-D dimethylamine on wood frog tadpoles in Saskatchewan.
Thesis Master of Science. University of Saskatchewan. Canada.
Kayhan, F.E., Turan, G T., Kaymak, G., Akbulut, C., & Ertug, N. D. Y. (2017). The 2, 4-D herbicidal effect on defense
enzyme activities and AChE levels in liver and gill tissues of Xiphoprus hellerii (Sworstail fish). Eltxir pollution, 110, 48560-48563.
Modesto, K.A. & Martinez, C. B. R., (2010). Roundup causes oxidative stress in liver and inhibits
acetylcholinesterase in muscle and brain of the fish Prochilodus lineatus. Chemosphere, 78, 294–299.
Mohamed, F.A.S. (2009). Histological studies on Tilapia zillii and Solea vulgaris from Lake Qurun, Egypt. World
Journal of fish and Marine science, 1(1), 29-39.
Nuankaew, C., Ngamsnae, P., Saowakoon, S., Nanthanawat, P. & Thanomsit, C. (2020). Toxicity of Paraquat on
Physiology, Behavior and Acetylcholinesterase (AChE) Expression in Nile tilapia (Oreochromis niloticus),
in: The 4th National Conference on Innovation for Learning and Invention 2020. pp. A16-A28.
OECD. Test Guidelines for the Chemicals - OECD. 2019. Test guideline No.203 Fish, acute toxicity testing. 1-23.
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.
Pollution Control Department (2010). 2, 4-dichlorophenoxyacetic acid (2, 4-D). Chulaprint. Bangkok. 84 p.
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 lowmolecular-mass drugs. Analytical Biochemistry, 407, 160–164.
Samanta, P. Rituparna, D.A.S., Sandipan, P.A.L., Aloke kumar, M., Senapati, T., Kole, D & Ghosh, A.R. (2018).
Toxicity assessment of agrochemical Almix in Heteroneustes fossils through histopathological alterations. Interdisciplinary Toxicology, 11(2), 129-147.
Sarikaya, R. & Selvi, M. (2005). Investigation of acute toxicity of (2, 4-dichlorophenoxy) acetic acid (2, 4-D) herbicide
on larvae and adult Nile tilapia (Oreochromis niloticus L.). Environmental Toxicology and Pharmacology, 20, 264-268.
Sarikaya, R., & Yilmaz, M. (2003). Investigation of acute toxicity and the effect of 2, 4-D (2,4
dichlorophenoxyacetic acid) herbicide on the behavior of the common carp (Cyprinus carpio L., 1758 ; Pisces, Cyprinidae). Chemosphere, 52, 195-201.
Thanomsit, C., Maprajuab, A., Saowakoon, A., Prasartkaew, W., Ocharoen, Y., Wattanakornsiri, A., Nanuam, J., &
Nanthanawat, P. (2018). Acetylcholinesterase (AChE): Potential Biomarker for evaluating Pesticide exposure on egg and tissue of golden apple snail (Pomecea canliculata) from Huai-Saneng Resevior, Surin Province, Thailand. Thai Journal of Agricultural science, 51(3), 104-117.
Thanomsit, C., Saowakoon, S., Wattanakornsiri, A., Nanuam, J., Prasatkaew, W., Nanthanawate, P., Monkolvai, P.
& Chalorcharoenying, W. (2020a). Glyphosate (Roundup): Fate, Toxicity Assessment and Adverse Effect on Aquatic Environment. Naresuan University Journal: Science and Technology, 28(1), 65-81.
Thanomsit, C., Nuankaew, C., Prasatkaew, W., Wattanakornsiri, A., Nanuam, J., Nanthanawat, P. & Kiatprasert, P.
(2020b). Adverse effects of chlorpyrifos and cypermethrin mixture on physiological alterations and
cholinesterase on Nile tilapia (Oreochromis niloticus). Thai journal of Agricultural science, 53(3), 134-148.
Thanomsit, C., Saowakoon, K., Wattanakornsiri, A., Khongchareonporn, N., Nanuam, J., Prasatkaew, W., &
Nanthanawat, P. (2020c) Acetylcholinesterase (AChE) Polyclonal Antibody from Hybrid catfish (C. macrocephalus × C. gariepinus): Specification, Sensitivity and Cross Reactivity. Biochemistry and Physiology Part C: Toxicology & Pharmacology, 108837.
Thanomsit, C., Wattanakornsiiri, A. & Nanthanawat, P. (2016a). Effect of glyphosate on fish behavior and
histological alteration of gills in Asian Sea bass (Lates calcarifer). Burapha Science Journal, 21 (2), 204-215.
Thanomsit, C., Wattanakornsiiri, A. & Nanthanawat, P. (2016b). Toxicity evaluation of glyphosate on mortality rate and histological alteration in Black Rice crab. Burapha Science Journal, 21(3), 151-165.
U.S. EPA. (United states Environment Protection Agency). 2005. Reregistration Eligibility Decision for 2, 4-D. USA.
321 p.
Vigario, A, F. & Saboia-Morais, S.M.T. (2014). Effect of the 2, 4-D herbicide on gills epithelia and liver of the fish
Poecilia vivipara. Brazilian Journal of Veterinary Research, 34(6), 523-528.
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2022-06-01
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