Exploring Lemna gibba thresholds to nutrient and chemical stressors: differential effects of triclosan on internal stoichiometry and nitrate uptake across a nitrogen:phosphorus gradient.

Nutrient enrichment often co-occurs with chemical stressors in aquatic ecosystems, but the impacts of these multiple stressors across nutrient gradients is poorly understood and not typically addressed in ecotoxicity studies of lower trophic level models. Moreover, laboratory assays performed to determine threshold responses of aquatic macrophytes to contaminants typically use growth and morphometric endpoints to establish threshold effects and seldom report other important functional responses of lower trophic levels. Using the aquatic macrophyte Lemna gibba, we examined influences of varying nitrogen (N) and phosphorus (P) levels in combination with triclosan, a widely used antimicrobial agent in consumer care products, on internal carbon (C):N:P and NO(3) (-) uptake kinetics. Triclosan modulated L. gibba tissue N and P content, and these stoichiometric responses for P-limited plants to triclosan exposure were more sensitive than growth endpoints employed in standardized phytotoxicity assays. Nitrate uptake capacities were also differentially inhibited by triclosan exposure according to external nutrient levels. Uptake rates for plants cultured and exposed under saturating N-levels were inhibited by more than threefold compared with N-limited plants. The results suggest that stoichiometric and nutrient uptake responses to chemical stressors provide useful information regarding adverse ecological thresholds not defined in standardized phytotoxicity assays with aquatic macrophytes. Our findings further indicate that site-specific impacts of chemicals associated with the wide ambient ranges of N and P typical of surface waters may be anticipated in lower trophic levels. Future studies should examine adverse effects of other stressors to these ecologically relevant endpoints, which may be useful in environmental assessment and management.

Influence of nitrogen and phosphorus concentrations and ratios on Lemna gibba growth responses to triclosan in laboratory and stream mesocosm experiments.

The effects of co-occurring nutrient and contaminant stressors are very likely to interact in aquatic systems, particularly at the level of primary producers. Site-specific nitrogen (N) and phosphorus (P) concentrations are often much lower and differ in relative availability than those used in nutrient-saturated laboratory assays for aquatic plants, which can introduce uncertainty in prospective ecological hazard and risk assessments. Because triclosan, an antimicrobial agent included in personal care products, potentially presents high relative risk among antimicrobial agents to aquatic plants and algae, we performed laboratory experiments with the model aquatic macrophyte Lemna gibba across a gradient of environmentally relevant N:P levels with and without triclosan co-exposure. Frond numbers (7 d) were significantly higher in N:P treatments of 16 and 23 but were lower in N:P of 937 and 2,500 treatments relative to standardized control media (N:P=3). When triclosan co-exposure occurred at high nutrient concentrations, frond number median effective concentration values at N:P 0.75, 3, and 16 were more than twofold lower than triclosan median effective concentration values in low nutrient media N:P ratios. However, a triclosan median effective concentration for frond number was twofold lower at N:P of 2,500 than at other N:P ratios in low concentration media. Influences of P enrichment on triclosan toxicity to L. gibba were further explored during a 14-d outdoor experimental stream mesocosm study. Effects of 2.6 and 20.8 microg L(-1) triclosan on L. gibba growth rates were more pronounced with increasing P treatment levels, which was generally consistent with our laboratory observations. Findings from these laboratory and field studies indicate that site-specific nutrient concentrations and ratios should be considered during assessments of primary producer responses to chemical stressors.