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Figure 1.

Workflow for the optimization and evaluation of earthworm responses to sub-lethal toxicity of Carbofuran.

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Table 1.

Compounds structurally identified from earthworm, Metaphire.posthuma by GC-MS.

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Figure 2.

PCA of optimization of extraction solvent system for global metabolite profiling of earthworm Metaphire posthuma A) scores plot, explaining the extraction efficiency of different solvent systems 1. 80% MeOH, 2. Pure MeOH, 3. MIPW, 4. MCW, 5. AMW. B) Loadings plot.

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Figure 3.

Coefficient of variation of metabolite features for different solvent systems extracted from earthworm Metaphire posthuma.

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Figure 4.

Clustered heat map of intracellular metabolites extracted using A) MCW B) AMW C) Pure MeOH D) MIPW E) 80% MeOH.

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Figure 5.

Principle component analysis (PCA) A) scores and B) loadings plots for 1.control earthworms and earthworms exposed to soil spiked with 2. Concentration of 0.1 mg/Kg, 3. Concentration of 0.3 mg/Kg, and 4. Concentration of 0.6 mg/Kg of carbofuran.

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Figure 6.

Predictive accuracy of the model discriminating carbofuran exposed and healthy control earthworms summarised using ROC curve analysis.

Area under the curve = 0.99.

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Figure 7.

Earthworm metabolite responses to carbofuran exposure 1) control group 2) concentration of 0.15 mg kg−1 exposed group 3) concentration of 0.3 mg kg−1 exposed group 4) concentration of 0.6 mg kg−1 exposed group.

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Table 2.

Key putatively identified metabolic perturbations from GC MS based metabolomic analyses of earthworms after exposed to carbofuran.

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Table 2 Expand