A Potential impact of glyphosate on morpho-behavioural changes, antioxidant system and lipid peroxidation products in the epigeic earthworm, Eudrilus eugeniae.
Published 2026-02-14
Keywords
- antioxidants,
- biomarkers,
- glyphosate,
- lipid peroxidation,
- oxidative stress
- earthworm ...More
Copyright (c) 2026

This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Abstract
The plants and animals in the soil ecosystem are negatively impacted by the regular use of chemical pesticides in agricultural areas. The usage of glyphosate-based herbicides (GBHs) and the discharge of glyphosate residues into the soil environment have raised serious concerns over time. This intentional release of pesticides into the soil environment highlights how important it is to keep an eye on the herbicides' ecotoxicological risk assessment and minimize any potential negative consequences. The current study examines the harmful effects of the herbicide glyphosate on the earthworm Eudrillus eugeniae from this perspective. In the epigeic earthworm, E. eugeniae, the non-enzymatic antioxidant glutathione (GSH) and the hepatic antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and glutathione peroxidase (GPx) were selected as biomarkers. The results demonstrated that the sublethal exposure to glyphosate had a significant effect on these biomarkers' activity. Lipid peroxidation (LPO) products, such as conjugate diene and malondialdehyde, were also altered by exposure. Through its interactions with antioxidant enzymes, the current study further demonstrated that glyphosate is a chemical with oxidative potential.
References
- Achuba, F. I. (2018). Protective influence of Elaesis guineensis leaf in diet on petroleum-mediated kidney damage in rats. Journal of Applied Pharmaceutical Sciences and Research. 7(2), 33-38.
- Ansaldo, M., Luquet, C. M., Evelson, P. A., Polo, J. M., & Llesuy, S. (2000). Antioxidant levels from different Antarctic fish caught around South Georgia Island and Shag rocks. Polar Biology. 23, 160-5.
- Banerjee, B. D., Seth, V., & Ahmed, R. S. (2001). Pesticide-induced oxidative stress: Perspectives and trends. Reviews on Environmental Health. 16,1–40.
- Benke, G. M., & Cheevar, K. C. (1974). The comparative toxicity, acetylcholinesterase action and metabolism of methyl parathion and parathion in sunfish and mice. Toxicology and Applied Pharmacology. 28, 97-109.
- Bohn, T., Cuhra, M., Traavik, T., Sanden, M., Fagan, J., & Primicerio R. (2013). Compositional differences in soybeans on the market: glyphosate accumulates in Roundup Ready GM soybeans. Food Chemistry. 153, 207–215.
- Booth, J., Boyland, E., & Sims, P. 1961. An enzyme from rat liver catalyzing conjugation with glutathione. Biochemical Journal. 79, 516-524.
- Bradbury, F. R., & Standan, H. (1959). Metabolism of benzene hexachloride by resistant house flies Musca domestica. Nature. 183, 983-984.
- Bradford, M. M., (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical. biochemistry, 72(1-2), 248-254.
- De Zwart, L. L., Venhorst, J., Groot, M., Commandeur, J. N. M., Hermanns, R. C. A., Meerman, J. H. N., Van Baar, B. L. L., & Vermeulen, N. P. E. (1997). Simultaneous determination of eight lipid peroxidation degradation products in urine of rats treated with carbon tetrachloride using gas chromatography with electron capture detection. Journal of Chromatography. 694: 277-288.
- Di Giulio, R. T., Washburn, P. C., Wenning, R. J., Winston, G. W., & Jewell, C. S. (1989). Biochemical responses in aquatic animals: A review of determinants of oxidative stress. Environmental Toxicology and Chemistry. 8, 1103–1123.
- Drake, H. L., & Horn, M. A. (2007). As the worm turns: the earthworm gut as a transient habitat for soil microbial biomes. Annual Review of Microbiology. 61,169–189.
- Duke, S. O., & Powles, S. B. (2008). Glyphosate: a once-in-a-century herbicide. Pest Management Science. 64(4), 319–325.
- Duke, S. O., & Powles, S. B., (2009). Glyphosate-resistant crops and weeds: now and in the future. The Journal of Agrobiotechnology Management. 12(3), 346–357.
- Duncan, D. B. (1955). Multiple range and multiple F- test. Biometrics 11, 1-42
- Guo, T. R., Zhang G. P, Zhou, M. X., Wu, F. B., & Chen, J. X. (2004). Effects of aluminum and cadmium toxicity on growth and antioxidant enzyme activities of two barley genotypes with different Al resistance. Plant and Soil. 258, 241–248.
- Habig, W. H., Pabst, M. J., & Jakoby, W. B. (1974). Glutathione-S-transferases: The first enzymatic step in mercapturic acid formation. Journal of Biololgical Chemistry. 249, 7130-7139.
- Hegedus, A., Erdei, S., & Horvath, G. (2001). Comparative studies of H2O2 detoxifying enzymes in green and greening barley seedlings under cadmium stress. Plant Science. 160,1085–1093.
- Jakoby, W. B., & Habig, W. H. (1980). Glutathione-S-transferase. In Enzymatic Basis of Detoxification (W. B. Jakoby. Ed.) Academic Press, New York. 2, 63-94.
- Johnson, M. K. (1966). Studies on glutathione-S-transferase of the rat. Biochemical Journal. 93, 3843.
- Kadar, E., Costa, V., Santos, R. S. (2006). Distribution of microessential (Fe, Cu, Zn) and toxic (Hg) metals in tissues of two nutritionally distinct hydrothermal shrimps. Science of the Total Environment. 71, 65-70.
- Kakkar, P., Das, B., & Viswanathan, P. N. (1984). A modified spectrophotometric assay of superoxide dismutase. Indian Journal of Biochemistry and Biophysics. 21(2), 130.
- Kelly, S., Havrilla, C., Brady, T. A., & Levin, E. (1998). Oxidative stress in toxicology: Established mammalian and emerging piscine model system. Environmental Health Perspectives. 106(7),375-384.
- Krea, S., Zaja, R., Calic, V., Terzic, S., Grubesic, M. S., & Ahel, N. (2007). Hepatic biomarker response to organic contamination in feral chub (Letrciscus cephalus) – Laboratory characterisation and field study in the Sava River, Croatia. Environmental. Toxicology and Chemistry. 26, 2620-26233.
- Kweki, G. R., Ichipi-Ifukor, P. C., & Asagba, S. O. (2018). High caffeine-containing Energy drink Induced Metabolic stress in Rats. Sok. Journal of Medical Laboratory Science. 3(3), 86-93
- Landrum, M., Canas, J. E., Coimbatore, G., Cobb, G. P., Jackson, W. A., Zhang, B., & Anderson, T. A., (2006). Effects of perchlorate on earthworm (Eisenia fetida) survival and reproductive success. Science of the Total Environment. 363, 237–244.
- Lawrence, R. A., & Burck, R. A. (1976). Glutathione peroxidase activity in selenium-deficient rat liver. Biochemical and Biophysical Research Communications. 71, 952-958.
- Livingstone, D. (2001). Contaminant-stimulated reactive oxygen species production and oxidative damage in aquatic organisms. Marine Pollution Bulletin. 42, 656-66.
- Livingstone, D., Lemaire, P., Matthews, A., Peters, L., Buke, D., & Law, R. (1993). Pro-oxidant, antioxidant and 7-ethoxyresorufin O-deethylase (EROD) activity responses in liver of dab (Limanda limanda) exposed to sediment contaminated with hydrocarbons and other chemicals. Marine Pollution Bulletin 26(11), 602-606.
- Lozowicka, B., Jankowska, M., Hrynko, I., & Kaczynski, P. (2016). Removal of 16 pesticide residues from strawberries by washing with tap and ozone water, ultrasonic cleaning and boiling. Environmental Monitoring & Assessment. 188(1),1–19.
- Lynn, H. B., & Kathryn, O. (2001). A comparison of biomarker responses in the earthworm Aporrectodea caliginosa to the organophosphorus insecticides Diazinon and chlorpyrifos. Environmental Toxicology and Chemistry. 20(11), 2494–2502.
- Maehly, A. C., & Chance, B. (1954). The assay of catalases and peroxidases. Wiley, New York, pp. 357–424.
- Matozzo, V., Fabrello, J., & Marin, M.G. (2020). The effects of glyphosate and its commercial formulations to marine invertebrates: a review. Journal of Marine and Science Engineering. 8(6), 399.
- McCarthy, J. F., & Shugart, L. R. (1990). Biomarkers of environmental contamination Boca Raton, FL: Lewis Publishers. 457.
- Neuhauser, E. F., Cukic, Z.V., Malecki, M. R., Loehr, R. C., & Durkin, P. R. (1995). Bioconcentration and biokinetics of heavy metals in the earthworm. Environmental Pollution. 89(3), 293-301.
- Niehaus, W. G., & Samuelsson, B. (1968). Formation of malonaldehyde from phospholipid arachidonate during microsomal lipid peroxidation. European Journal of biochemistry. 6(1), 126-130.
- Oakes, K. D., & Van Der Kraak, G. J. (2003). Utility of the TBARS assay in detecting oxidative stress in white sucker (Catostomus commersoni) populations exposed to pulp mill effluent. Aquatic Toxicology, 63(4), 447-463.
- Oakes, K. D., McMaster, M. E., & Van Der Kraak, G. J. (2004). Oxidative stress responses in longnose sucker (Catostomus catostomus) exposed to pulp and paper mill and municipal sewage effluents. Aquatic Toxicology, 67(3), 255-271.
- OECD (Organization for Economic Cooperation and Development). (1984). OECD Guidelines for Testing of Chemicals. Section 2, Effects on Biotic Systems: Earthworm Acute Toxicity Tests. Paris, France.
- Orbea, A., Fahimi, H., & Cajaraville, M. (2000). Immunolocalization of four antioxidant enzymes in digestive glands of mollusks and crustaceans and fish liver. Histochemistry and Cell Biology. 114, 393-404.
- Ozkara, A., Akyıl, D., & Konuk, M. (2016). In Environmental health risk-hazardous factors to living species. Pesticides, environmental pollution, and health. 8(6), 399.
- Padmini, E., Vijaya Geetha, B., & Usha Rani, M. (2008). Liver oxidative stress of grey mullet, Mugil cephalus, present seasonal variations in Ennore estuary. Brazilian Journal of Medical and Biological Research. 41(11), 951-955.
- Pastore, A., Piemonte, F., Locatelli, M., Russo, A. L., Gaeta, L. M., Tozzi, G., & Federici, G. (2003). Determination of blood total, reduced, and oxidized glutathione in pediatric subjects. Clinical Chemistry. 47(8), 1467–1469.
- Plummer, J. L., Smith, B. R., Sies, H., & Bend, J. R. (1981). Chemical depletion of glutathione in viro. In Methods in Enzymology (W. J. Jakoby, Ed.) Academic Press, New York. 77, 50-51.
- Recknagel, R. O., & Ghoshal, A. K. (1966). Lipoperoxidation of rat liver microsomal lipids induced by carbon tetrachloride. Nature. 210(5041), 1162-1163.
- Reinecke, S. A., & Reinecke, A. J. (2007). The impact of organophosphate pesticides in orchards on earthworms in the Western Cape, South Africa. Ecotoxicological and Environmental Safety. 66(2), 244–251.
- Ribera, D., Narbonne, J. F., Arnaud, C., & Saint-Denis, M. (2001). Biochemical responses of the earthworm Eisenia fetida andrei exposed to contaminated artificial soil, effects of carbaryl. Soil Biology and Biochemistry. 33, 1123–1130.
- Salisbury, F. B., & Ross, C. W. (1994). Fisiologia Vegetal. Grupo Editorial Iberoamerica S.A., Mexico.
- Salvio, C., Menone, M. L., Rafael, S., Iturburu, F. G., & Manetti, P. L. (2016). Survival, reproduction, avoidance behavior and oxidative stress biomarkers in the earthworm Octolasion cyaneum exposed to glyphosate. Bulletin of environmental contamination and toxicology. 96, 314-319.
- Sanchez, W., Palluel, O., Meunier, L., Coquery, M., Porcher, J. M., & Ait-Aissa, S. (2005). Copper-induced oxidative stress in three-spined stickleback: relationship with hepatic metal levels. Environmental toxicology and pharmacology. 19(1), 177-183.
- Sanchez-Hernandez, J. C. (2006). Earthworm biomarkers in ecological risk assessment. Reviews of Environmental Contamination and Toxicology. 188, 85–126.
- Shaner, D. (2006). An overview of glyphosate mode of action: why is it such a great herbicide? North Central Weed. Science Society. 61, 94.
- Sies, H. (1986). Biochemistry of oxidative stress. Angewandte Chemie International Edition in English, 25(12), 1058-1071.
- Suryasikha Samal, S. S., Sunanda Sahoo, S. S., & Mishra, C. S. K. (2017). Morpho-histological and enzymatic alterations in earthworms Drawida willsi and Lampito mauritii exposed to urea, phosphogypsum and paper mill sludge. 762-776.
- Tjalkens, R. B., Valeriojr, I. G., Awasthi, Y. C., & Petersen, D. R. (1998). Association of GST induction and lipid peroxides in two inbred strains of mice subjected to chronic iron overload. Toxicology and Applied Pharmacology. 151, 174-81.
- Uner, N., Ozcan-Oruc, E., Canli, M., & Sevgiler, Y. (2001). Effects of cypermethrin on antioxidant enzyme activities and lipid peroxidation in liver and kidney of the freshwater fish, Oreochromis niloticus and Cyprinus carpio (L.). Bulletin of Environmental Contamination of Toxicology. 67,657–664.
- Wang, J. H., Zhu, L. S., Liu, W., Wang, J., & Xie, H., (2012). Biochemical responses of earthworm (Eisenia foetida) to the pesticides chlorpyrifos and fenvalerate. Toxicology Mechanisms and Methods, 22(3), 236-241.
- Wang, K., Mu, X., Qi S., Chai, T., Pang, S., Yang, Y., & Jiang, J. (2015). Toxicity of a neonicotinoid insecticide, guadipyr, in earthworm (Eisenia fetida). Ecotoxicology and Environmental Safety. 114, 17–22.
- Wiegand, C., Pehkonen, S., Akkanen, J., Penttinen, O. P., & Kukkonen, J. V. K. (2007). Bioaccumulation of paraquat by Lumbriculus variegatus in the presence of dissolved natural organic matter and impact on energy costs, biotransformation and antioxidative enzymes. Chemosphere. 66, 558–566.
- Winston, G. W., & Di Giulio, R. T. 1991. Prooxidant and antioxidant mechanisms in aquatic organisms. Aquatic Toxicology. 19, 137-161.
- Xu, Z., Wang, Z., Li, J. J., Chen, C., Zhang, P. C., Dong, L., & Wang, Z. L. (2013). Protective effects of selenium on oxidative damage and oxidative stress related gene expression in rat liver under chronic poisoning of arsenic. Food and chemical toxicology, 58, 1-7.
- Yang, Y., Cheng, J. Z., Singhal, S. S., Saini, M., Pandya, U., & Awasthi, S. (2001). Role of glutathione S-transferases in protection against lipid peroxidation. Journal of Biological Chemistry. 276: 220-230.