Bioaccumulation of sediment-associated substituted phenylamine antioxidants in Tubifex tubifex and Lampsilis siliquoidea.

Substituted phenylamine antioxidants (SPAs) are additives in a variety of commercial polymers (e.g., lubricants, plastics, etc.). Based on their physicochemical properties, if SPAs were to enter an aquatic system, they would likely partition into sediment and have the capacity to bioaccumulate in biota. This study investigated the potential of four sediment-associated SPAs, diphenylamine (DPA), N-phenyl-1-naphthalene (PNA), N-(1,3-dimethylbutyl)-N'-phenyl-1,4-phenylenediamine (DPPDA), and 4,4'-methylene-bis[N-sec-butylaniline] (MBA) to accumulate in the tissues of freshwater mussels (Lampsilis siliquoidea) and oligochaete worms (Tubifex tubifex). Mussels and worms were exposed to sediment spiked with individual SPAs for 28 d. The concentration of SPAs was measured in the gill, gonad, and remaining viscera of the mussels and entire body of the worms. The majority of biota-sediment accumulation factors (28-d BSAFs) for the different tissues of mussels were < 1. The highest concentrations of SPAs were consistently observed in the gill tissue of mussels relative to the gonad and viscera. The 28-d BSAFs for DPPDA and MBA for worms were < 1, and for DPA and PNA, they ranged from 0.38-2.13 and 1.54-33.24, respectively. The higher 28-d BSAFs observed for worms compared to mussels were likely because worms are endobenthic and feed on sediment-associated organic matter. PNA and DPPDA have similar octanol-water partition coefficients (Kow) but greater 28-d BSAFs were observed for PNA compared to DPPDA for both species. This observation provides evidence that biota may be able to metabolize and/or excrete SPAs with similar physicochemical properties at considerably different rates. The 28-d BSAFs observed for sediment-associated SPAs are lower than those typically required for a chemical to be classified as bioaccumulative.

Effect of substituted phenylamine antioxidants on three life stages of the freshwater mussel Lampsilis siliquoidea.

Substituted phenylamines (SPAs) are incorporated into a variety of consumer products (e.g., polymers, lubricants) in order to increase the lifespan of the products by acting as a primary antioxidant. Based on their physicochemical properties, if SPAs were to enter the aquatic environment, they would likely partition into sediment. No studies to date have investigated the effect of sediment-associated SPAs on aquatic organisms. The current study examined the effect of four SPAs (diphenylamine (DPA); N-phenyl-1-napthylamine (PNA); N-(1,3-dimethylbutyl)-N'-phenyl-1,4-phenylenediamine (DPPDA); 4,4'-methylene-bis[N-sec-butylaniline] (MBA)) on three different life stages of the freshwater mussel, Lampsilis siliquoidea. The viability of larvae (glochidia) of L. siliquoidea and Lampsilis fasciola was assessed after 48 h of exposure to SPAs in water. The 48-h EC50s for glochidia viability of L. siliquoidea were 5951, 606, 439, and 258 µg/L for DPA, PNA, DPPDA, and MBA, respectively, and 7946, 591, 137, and 47 µg/L, respectively, for L. fasciola. Juvenile (7-15 months) and adult L. siliquoidea were exposed to sediment-associated SPAs for 28 d. LC50s for juvenile mussels were 18, 55, 62, and 109 µg/g dry weight (dw) of sediment for DPA, PNA, DPPDA, and MBA, respectively. Adult mussels were exposed to sub-lethal concentrations of sediment-associated SPAs in order to investigate reactive oxygen species (ROS), lipid peroxidation and total glutathione in the gill, gonad, and digestive gland tissue, and viability and DNA damage in hemocytes. No significant concentration-dependent trend in any of these biochemical and cellular endpoints relative to the concentration of sediment-associated SPAs was observed in any tissues. Investigations into the concentration of SPAs in the aquatic environment are required before a conclusion can be made on whether these compounds pose a hazard to the different life stages of freshwater mussels.

Effects of in situ exposure to tritiated natural environments: A multi-biomarker approach using the fathead minnow, Pimephales promelas.

Aquatic ecosystems are chronically exposed to radionuclides as well as other pollutants. Increased concentrations of pollutants in aquatic environments can present a risk to exposed organisms, including fish. The goal of this study was to characterize the effects of tritium, in the context of natural environments, on the health of fathead minnow, Pimephales promelas. Fish were exposed to tritium (activity concentrations ranging from 2 to 23,000Bq/L) and also to various concentrations of several metals to replicate multiple-stressor environments. Fish were exposed for 60days, then transferred to the tritium background site where they stayed for another 60days. Tritium, in the forms of tritiated water (HTO) and organically bound tritium (OBT), and a series of fish health indicators were measured in fish tissues at seven time points throughout the 120days required to complete the exposure and the depuration phases. Results showed effects of environmental exposure following the increase of tritium activity and metals concentrations in water. The internal dose rates of tritium, estimated from tissue HTO and OBT activity concentrations, were consistently low (maximum of 0.2µGy/h) compared to levels at which population effects may be expected (>100µGy/h) and no effects were observed on survival, fish condition, gonado-somatic, hepato-somatic, spleno-somatic and metabolic indices (RNA/DNA, proteins/DNA and protein carbonylation (in gonads and kidneys)). Using multivariate analyses, we showed that several biomarkers (DNA damages, MN frequency, gamma-H2AX, SFA/MUFA ratios, lysosomal membrane integrity, AChE, SOD, phagocytosis and esterase activities) were exclusively correlated with fish tritium internal dose rate, showing that tritium induced genotoxicity, DNA repair activity, changes in fatty acid composition, and immune, neural and antioxidant responses. Some biomarkers were responding to the presence of metals, but overall, more biomarkers were linked to internalized tritium. The results are discussed in the context of multiple stressors involving metals and tritium.

Variation in the toxicity of sediment-associated substituted phenylamine antioxidants to an epibenthic (Hyalella azteca) and endobenthic (Tubifex tubifex) invertebrate.

Substituted phenylamine antioxidants (SPAs) are produced in relatively high volumes and used in a range of applications (e.g., rubber, polyurethane); however, little is known about their toxicity to aquatic biota. Therefore, current study examined the effects of chronic exposure (28 d) to four sediment-associated SPAs on epibenthic (Hyalella azteca) and endobenthic (Tubifex tubifex) organisms. In addition, acute (96-h), water-only exposures were conducted with H. azteca. Mortality, growth and biomass production were assessed in juvenile H. azteca exposed to diphenylamine (DPA), N-phenyl-1-napthylamine (PNA), N-(1,3-dimethylbutyl)-N'-phenyl-1,4-phenylenediamine (DPPDA), or 4,4'-methylene-bis[N-sec-butylaniline] (MBA). Mortality of adult T. tubifex and reproduction were assessed following exposure to the four SPAs. The 96-h LC50s for juvenile H. azteca were 1443, 109, 250, and >22 µg/L and 28-d LC50s were 22, 99, 135, and >403 µg/g dry weight (dw) for DPA, PNA, DPPDA, and MBA, respectively. Reproductive endpoints for T. tubifex (EC50s for production of juveniles > 500 µm: 15, 9, 4, 3.6 µg/g dw, for DPA, PNA, DPPDA, and MBA, respectively) were an order of magnitude more sensitive than endpoints for juvenile H. azteca and mortality of adult worms. The variation in toxicity across the four SPAs was likely related to the bioavailability of the sediment-associated chemicals, which was determined by the chemical properties of the SPAs (e.g., solubility in water, Koc). The variation in the sensitivity between the two species was likely due to differences in the magnitude of exposure, which is a function of the life histories of the epibenthic amphipod and the endobenthic worm. The data generated from this study will support effect characterization for ecological risk assessment.