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  4. The exposure of ecologically critical invertebrates to pharmaceutically active compounds in aquatic environments has been one of the major concerns over the past decade, which also adds serious risk to the aquatic ecosystem. However, the metabolic level perturbations in invertebrates in response to sub-lethal doses of pharmaceuticals are still rarely studied, especially in the marine coastal environment. In this study, the diclofenac regulation of southern hard clam Mercenaria campechiensis metabolites at different time points and concentrations was investigated using NMR-based metabolomics. As a result, clam metabolic profile perturbations were observed under both low and high concentrations of diclofenac exposure in one week according to principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA); however, the potential influenced metabolic pathways were distinctly different. The low-concentration group showed significant taurine upregulation, which indicated self-protection from osmotic stress. However, the metabolites succinate, alanine, and glutamate were significantly upregulated at the relatively high concentration of diclofenac, which was a sign of anaerobic activities. The metabolic profile perturbations in week 2 showed high similarity in both low- and high-concentration groups, and the osmotic protectants betaine and taurine were significantly downregulated. The study indicated the early markers of diclofenac exposure in M. campechiensis , which provided pioneering results for monitoring the toxicity of pharmaceuticals to marine coastal water. 
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  5. Abstract

    Nutrient enrichment often alters the biomass and species composition of plant communities, but the extent to which these changes are reversible after the cessation of nutrient addition is not well‐understood. Our 22‐year experiment (15 years for nutrient addition and 7 years for recovery), conducted in an alpine meadow, showed that soil nitrogen concentration and pH recovered rapidly after cessation of nutrient addition. However, this was not accompanied by a full recovery of plant community composition. An incomplete recovery in plant diversity and a directional shift in species composition from grass dominance to forb dominance were observed 7 years after the nutrient addition ended. Strikingy, the historically dominant sedges with low germination rate and slow growth rate and nitrogen‐fixing legumes with low germination rate were unable to re‐establish after nutrient addition ceased. By contrast, rapid recovery of aboveground biomass was observed after nutrient cessation as the increase in forb biomass only partially compensated for the decline in grass biomass. These results indicate that anthropogenic nutrient input can have long‐lasting effects on the structure, but not the soil chemistry and plant biomass, of grassland communities, and that the recovery of soil chemical properties and plant biomass does not necessarily guarantee the restoration of plant community structure. These findings have important implications for the management and recovery of grassland communities, many of which are experiencing alterations in resource input.

     
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