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Metabolomics reveals increased toxicity in earthworms (Eisenia fetida) of the transformation product methyl-triclosan

ABSTRACT: The effects of the antimicrobial triclosan (TCS) and its transformation product methyl-triclosan (MTCS) on the earthworm Eisenia fetida were investigated using GC-MS metabolomics. TCS is ubiquitous in sewage sludge, but a large proportion is transformed into MTCS during wastewater treatment and in soil when sewage sludge is applied to land. Our objective was to determine if earthworms exposed to ng/g to μg/g concentrations of TCS or MTCS exhibit toxic effects, and to identify the toxic mode of action of each compound. Ten individual earthworm replicates in 10 g worm bedding were exposed to 0, 0.25, 1, 4, 16, or 64 μg/g of either TCS or MTCS (120 experimental units) for 14 days. No mortalities were observed. All MTCS exposed worms had an instantaneous growth rate (IGR) over two times higher than the control during the study, but there was no effect of increasing concentration. Succinic acid was elevated relative to the control at concentrations ≥ 0.25 μg/g and glucose was elevated at 1 μg/g. There was separation from the control at all concentrations except 4 μg/g using Principal Components Analysis. Glucose, palmitic acid, and IGR contributed most strongly to the separation. Discriminant analysis with succinic acid, glucose, and IGR as variables showed a clear separation at all concentrations from the control along Canonical 1. Disruption of energy metabolism was hypothesized as a possible mode of action for MTCS.

INSTRUMENT(S): 5975C Series GC/MSD (Agilent)

SUBMITTER: Joseph Gillis

PROVIDER: MTBLS532 | MetaboLights | 2018-09-17

REPOSITORIES: MetaboLights

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Project description:Triclocarban (TCC) is a widely used antimicrobial agent that is routinely detected in surface waters. The present study was designed to examine TCC’s efficacy and mode of action as a reproductive toxicant in fish. Reproductively mature Pimephales promelas were continuously exposed to either 1 or 5 μg TCC/L, 0.5 μg 17β-trenbolone (TRB)/L or a mixture (MIX) of 5 μg TCC and 0.5 μg TRB/L for 22 d and a variety of reproductive and endocrine-related endpoints were examined. The data were evaluated to answer several key questions: first, whether exposure to TCC could be linked with an ecologically-relevant adverse outcome, i.e., impaired reproduction; second, whether the present study provided additional support for augmentation of androgen action as an endocrine-disrupting mode of action for TCC; and third, whether there were any novel and/or plausible linkages between changes in the ovarian transcriptome and the apical responses observed in females that could support adverse outcome pathway development and/or further hypothesis-driven testing. Fathead minnows were randomly paired (one male, one female) and placed in tanks at a density of two pairs per tank with six replicate tanks per treatment. Pairs were separated by a water permeable mesh divider and each pair had its own breeding substrate. The fish were held in the test system, receiving UV-treated, filtered Lake Superior (control) water only, for a 21 d acclimation period during which the fecundity and fertility of each pair were assessed daily. After 21 d, exposures were initiated with pairs that had spawned successfully during acclimation. Fish were exposed continuously to target test concentrations of 0 (Lake Superior control), TCC (1 and 5 µg/L), TRB (0.5 μg/L), or a mixture of the two chemicals (5 μg TCC and 0.5 μg TRB/L) for 22 d. Water concentrations of TCC and TRB were determined in stock solutions and all exposure tanks every two to three days. General water quality characteristics (mean ± SD) measured regularly over the course of the Experiment were: temperature, 24.8 ± 0.7 oC; pH, 7.87 ± 0.04; dissolved oxygen, 6.8 ± 0.35 mg/L. Ovary RNA samples from twenty four fish (n=5 for all treatments except TRB alone; n=4) were selected for microarray analysis.

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Metabolomics reveals increased toxicity in earthworms (Eisenia fetida) of the transformation product methyl-triclosan

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