Coelomic fluid: a complimentary biological medium to assess sub-lethal endosulfan exposure using ¹H NMR-based earthworm metabolomics.

Endosulfan is an environmentally persistent pesticide and has been shown to be genotoxic, neurotoxic and carcinogenic to surrounding organisms. Earthworms are widely used in environmental metabolomic studies to assess soil ecotoxicity. Previous nuclear magnetic resonance (NMR)-based metabolomic studies have analyzed earthworm tissue extracts after exposure to endosulfan and identified some key metabolic indicators that can be used as biomarkers of stress. However, some metabolites may have been masked due to overlap with other metabolites in the tissue extract. Therefore, in this study, the coelomic fluid (CF) and the tissue extract of the earthworm, Eisenia fetida, were both investigated using ¹H NMR-based metabolomics to analyze their metabolic profile in response to endosulfan exposure at three sub-lethal (below LC₅₀) concentrations. Principal component analysis determined the earthworm CF and earthworm tissue extract to both have significant separation between the exposed and control at the two highest sub-lethal endosulfan exposures (1.0 and 2.0 μg cm⁻²). Alanine, glycine, malate, α-ketoglutarate, succinate, betaine, myo-inositol, lactate and spermidine in the earthworm CF and alanine, glutamine, fumarate, glutamate, maltose, melibiose, ATP and lactate in earthworm tissue extract were all detected as having significant fluctuations after endosulfan exposure. An increase in ATP production was detected by the increase activity in the citric acid cycle and by anaerobic metabolism. A significant decrease in the polyamine, spermidine after endosulfan exposure describes an apoptotic mode of protection which correlates to a previous endosulfan exposure study where DNA damage has been reported. This study highlights that earthworm CF is a complementary biological medium to tissue extracts and can be helpful to better understand the toxic mode of action of contaminants at sub-lethal levels in the environment.