Exposures to a selective serotonin reuptake inhibitor (SSRI), sertraline hydrochloride, over multiple generations: changes in life history traits in Ceriodaphnia dubia.

Selective serotonin reuptake inhibitors (SSRIs) have been reported to range from low parts per trillion to parts per billion levels in surface waters, wastewater effluents, and sediments. These low levels have led to concern for their potential long-term risks to the survival, growth, and reproduction of aquatic organisms. We investigated the acute and chronic effects of sertraline on the life history traits of Ceriodaphnia dubia over the course of three generations under environmentally realistic concentrations. Acute toxicity of sertraline in C. dubia offspring resulted in a 48h median effective concentration of 126µgL(-1). Under chronic exposure, the lowest concentration to affect fecundity and growth was at 53.4µgL(-1) in the first two generations. These parameters become more sensitive during the third generation where the LOEC was 4.8µgL(-1). The median effective concentrations (EC50) for the number of offspring per female, offspring body size, and dry weight were 17.2, 21.2, and 26.2µgL(-1), respectively. Endpoints measured in this study demonstrate that chronic exposure of C. dubia to sertraline leads to effects that occur at concentrations only an order of magnitude higher than predicted environmental concentrations. However, this study also demonstrates that multigenerational effects should be considered in chronic exposure studies because standard toxicity tests do not account for increases in sensitivity in successive generations to toxicants.

Chronic effects of carbamazepine on life-history strategies of Ceriodaphnia dubia in three successive generations.

Trace quantities of pharmaceuticals are continuously being discharged into the environment through domestic and industrial wastewater effluents, causing concern among scientists and regulators regarding potential long-term impacts on aquatic ecosystems. These compounds and their metabolites are constantly interacting with organisms at various life-cycle stages and may differentially influence the development of embryonic, larval, juvenile, and adult stages. To understand the possible cumulative effects of exposure to carbamazepine (CBZ), a multigenerational approach was taken in which survival, reproduction, respiration, growth, brood size, and biomass of Ceriodaphnia dubia were assessed at sublethal concentrations over the course of three successive generations. CBZ exposure significantly decreased fecundity at 196.7 µg/L in the F0 and F1 generations over 2 weeks and acclimatized at 264.6 µg/L in the F2 generation. Similarly, a significant decrease of neonate dry weight was observed at the 196.7 µg/L CBZ treatment in the F1 generation, and it acclimatized at 264.6 µg/L treatment level in the F2 generation. Median time to first brood release was significantly delayed at 264.6 µg/L in the F2 generation, indicating slower maturation. Results over three successive generations are not different than what one would obtain by testing simply the F0 generation. Furthermore, the effects measured were observed at concentrations two orders of magnitude higher than are environmentally relevant, and it is unlikely that CBZ poses a substantial risk to the environment regarding the end points measured in this study. However, additional research through laboratory and field multigenerational studies may be required to understand the overall risk of CBZ to other nontarget organisms.

Snail bioaccumulation of triclocarban, triclosan, and methyltriclosan in a North Texas, USA, stream affected by wastewater treatment plant runoff.

Grazing by freshwater snails promotes nutrient turnover in algal communities. Grazed algal compartments may include antimicrobial agents and metabolites, such as triclocarban (TCC), triclosan (TCS), and methyltriclosan (MTCS), which are incompletely removed by wastewater treatment plant (WWTP) processing. The present study quantifies snail bioaccumulation factors (BAFs) for TCC, TCS, and MTCS at the outfall of Pecan Creek (TX, USA), the receiving stream for the city of Denton (TX, USA) WWTP. Helisoma trivolvis (Say) is ubiquitous and thrives under standard laboratory conditions, leading to its choice for this bioaccumulation study in conjunction with Cladophora spp. Along with providing substrate for epiphytic growth, Cladophora spp. provide a source of food and shelter for H. trivolvis. After being caged for two weeks, algae and snails were collected from the WWTP outfall, along with water-column samples, and analyzed by isotope dilution gas chromatography-mass spectrometry for TCS and MTCS and by liquid chromatography-mass spectrometry for TCC. Algal and snail samples were analyzed before exposure and found to be below practical quantitation limits for all antimicrobial agents. Triclocarban, TCS, and MTCS in water samples were at low-ppt concentrations (40-200 ng/L). Triclocarban, TCS, and MTCS were elevated to low-ppb concentrations (50-300 ng/g fresh wt) in caged snail samples and elevated to low-ppb concentrations (50-400 ng/g fresh wt) in caged algal samples. Resulting snail and algal BAFs were approximately three orders of magnitude, which supports rapid bioaccumulation among algae and adult caged snails at this receiving stream outfall. The results further support TCC, TCS, and MTCS as good candidate marker compounds for evaluation of environmental distribution of trace WWTP contaminants.

Algal bioaccumulation of triclocarban, triclosan, and methyl-triclosan in a North Texas wastewater treatment plant receiving stream.

Algae comprise the greatest abundance of plant biomass in aquatic environments and are a logical choice for aquatic toxicological studies, yet have been underutilized in this capacity. The lipid content of many algal species provides a point of entry for trophic transfer of lipophilic organic contaminants. Triclosan (TCS) and triclocarban (TCC), widely used antimicrobial agents found in numerous consumer products, are incompletely removed by wastewater treatment plant (WWTP) processing. Methyl-triclosan (M-TCS) is a metabolite of TCS more lipophilic than the parent compound. The focus of this study was to quantify algal bioaccumulation factors (BAFs) for TCS, M-TCS, and TCC in Pecan Creek, the receiving stream for the City of Denton, Texas WWTP. The complex algal compartment was field identified for collection and verified by laboratory microscopic description as being comprised of mostly filamentous algae (Cladophora spp.) and varying inconsequential levels of epiphytic diatoms and biofilm. Algae and water column samples were collected from the WWTP outfall, an upstream site, and two downstream sites and analysed by isotope dilution gas chromatography/mass spectrometry (GC/MS) for TCS and M-TCS and liquid chromatography/mass spectrometry (LC/MS) for TCC. TCS, M-TCS, and TCC in Pecan Creek water samples taken at and downstream from the WWTP were at low ppt concentrations of 50-200 ng l(-1) and were elevated to low ppb concentrations of 50-400 ng g(-1) fresh weight in algae collected from these stations. The resulting BAFs were approximately three orders of magnitude. TCS, M-TCS and TCC appear to be good candidate marker compounds for evaluation of environmental distribution of trace WWTP contaminants. Residue analysis of filamentous algal species typically occurring in receiving streams below WWTP discharges is a readily obtained indicator of the relative bioaccumulative potential of these trace contaminants.

Solid phase microextraction of aminodinitrotoluenes in tissue.

Tubifex tubifex metabolizes 2,4,6-trinitrotoluene (TNT) to 2-amino-4,6-dinitrotoluene (2ADNT) and 4-amino-2,6-dinitrotoluene (4ADNT). Elimination rates of metabolically-generated ADNTs are low compared to ADNTs absorbed directly from water, suggesting that metabolically-generated ADNTs may be bound or sequestered within tissue and therefore less available for elimination. A solid phase microextraction (SPME) technique was used to extract ADNTs from T. tubifex tissue to investigate the recalcitrance of metabolically-generated ADNTs. As SPME is a gentle, non-depletive, equilibrium sampling technique useful for measuring "available" organic compounds, we hypothesized that metabolically-generated ADNTs would be less extractable than absorbed ADNTs. T. tubifex were exposed to two scenarios to generate tissues containing absorbed ADNTs and metabolically-generated ADNTs. Tissue was then homogenized in a neutral buffer solution. Polyacrylate-coated (PA) SPME fibers were deployed and agitated in tissue homogenates to measure available ADNTs. Extractability of ADNTs from tissue containing metabolically-generated ADNTs was significantly less than expected: 50-60% based on the theoretical fiber-water partition ratio. Extractability of absorbed ADNTs was significantly higher (81-90%), and not significantly different than expected. The lower SPME extractability of metabolically-generated ADNTs may stem from the unavailability of metabolically-generated ADNTs sequestered in tissue or bound to tissue macromolecules during metabolism of TNT to ADNT. Tissue extractions using SPMEs may be able to estimate bound organic residues in tissue and serve to indicate the toxicological bioavailability of tissue-associated organic compounds.

Solid-phase microextraction for predicting the bioavailability of 2,4,6-trinitrotoluene and its primary transformation products in sediment and water.

Disposable solid-phase microextraction fibers (SPMEs) were used to measure the availability of 2,4,6-trinitrotoluene (TNT) and its two primary transformation products, 2-amino-4,6-dinitrotoluene (2ADNT) and 4-amino-2,6-dinitrotoluene (4ADNT). The SPMEs (85-microm polyacrylate) and sediment-dwelling oligochaetes (Tubifex tubifex) were exposed to TNT-spiked sediment, to TNT-spiked sediment amended with activated carbon, and to TNT-, 2ADNT-, and 4ADNT-spiked water. Sediment concentration was a poor predictor of bioavailability in unamended and carbon-amended sediments (r2 = 0.14-0.73) The activated carbon amendment reduced the bioavailability of compounds in carbon-amended sediment, causing the relationships between Tubifex concentrations and sediment concentrations to differ significantly between unamended and carbon-amended sediment for all compounds. In contrast, SPME TNT concentrations predicted Tubifex TNT concentrations (r2 = 0.54-0.79). and regression models did not differ significantly among the three TNT-spiked matrices. The SPME 2ADNT and 4ADNT concentrations also were predictive of Tubifex 2ADNT and 4ADNT concentrations (r2 = 0.44-0.90). Relationships between Tubifex concentrations and SPME concentrations were the same between unamended and carbon-amended TNT-spiked sediments for 2ADNT and 4ADNT; however, the relationship in sediment (pooled data) differed from the relationship found in 2ADNT- and 4-ADNT-spiked water. The SPMEs provided carbon amendment-independent measures of ADNT availability in sediment and matrix-independent measures of TNT availability among the three matrices. The SPMEs show promise for predicting bioavailable organic compounds in sediment and water.

A comparison of chronic cadmium effects on Hyalella azteca in effluent-dominated stream mesocosms to similar laboratory exposures in effluent and reconstituted hard water.

Laboratory single-species toxicity tests are used to assess the effects of contaminants on aquatic biota. Questions remain as to how accurately these toxicity tests predict site-specific bioavailability and chronic effects of metals, particularly in streams that are effluent-dominated or dependent on effluent discharge for flow. Concurrent 42-d Hyalella azteca exposures were performed with cadmium and final treated municipal effluent in the laboratory and at the University of North Texas Stream Research Facility (Denton, TX, USA), a series of outdoor lotic mesocosms. An additional 42-d laboratory test was conducted with H. azteca to evaluate Cd toxicity in reconstituted hard water (RHW). Endpoints included Cd body burden, survival, growth, and reproduction. Calculated average bioaccumulation factors were: 2,581 (stream mesocosm test) < 3,626 (laboratory effluent) < 7,382 (laboratory RHW). The 42-d survival lowest-observed-effect concentrations (LOECs) were 0.94, 4.53, and 22.97 microg/L for the laboratory RHW, laboratory effluent, and stream mesocosm exposures, respectively. Baseline growth (dry wt) and reproduction (young female(-1)) among the three exposures followed the relationship: Stream mesocosms > laboratory effluent > laboratory RHW. Differences among response variables in the three tests likely resulted from increased food sources and decreased Cd bioavailability in lotic mesocosms. Our results demonstrate that laboratory toxicity tests may overestimate chronic toxicity responses of H. azteca to Cd in effluent-dominated streams.

Comparison of in vitro and in vivo bioassays for estrogenicity in effluent from North American municipal wastewater facilities.

Attempts to better understand causal factors affecting estrogenicity in municipal wastewater have primarily focused on analytical evaluation of specific chemical estrogens and the use of estrogen receptor (ER) based in vitro assays. To compare analytical,in vitro, and in vivo assays for estrogenicity, wastewater from four New York and one Texas municipal wastewater facilities was evaluated for estrogenic activity using the yeast estrogen screen assay (YES) and an in vivo fish vitellogenin (VTG) assay. Estrogenic activity, as measured by the YES assay, was observed in methanol and/or methylene chloride eluents from C18 extracts in two of the New York treatment facilities and the Texas facility. Estradiol equivalents for the YES assay data ranged from

Aquatic ecotoxicology of fluoxetine.

Recent studies indicate that the pharmaceutical fluoxetine, a selective serotonin reuptake inhibitor, is discharged in municipal wastewater treatment plant effluents to surface waters. Few data on environmental fluoxetine exposure and hazard to aquatic life are currently available in the literature. Here, we summarize information on fluoxetine detection in surface waters and review research on single-species toxicity test, Japanese medaka (Oryzias latipes) reproduction and endocrine function, and freshwater mesocosm community responses to fluoxetine exposure. Based on results from our studies and calculations of expected introduction concentrations, we also provide a preliminary aquatic risk characterization for fluoxetine. If standard toxicity test responses and a hazard quotient risk characterization approach are solely considered, little risk of fluoxetine exposure may be expected to aquatic life. However, our findings indicate that: (1) the magnitude, duration and frequency of fluoxetine exposure in aquatic systems requires further investigation; (2) mechanistic toxicity of fluoxetine in non-target biota, including behavioral responses, are clearly not understood; and (3) an assessment of environmentally relevant fluoxetine concentrations is needed to characterize ecological community responses.

Response of Ceriodaphnia dubia to ionic silver: discrepancies among model predictions, measured concentrations and mortality.

Silver thiosulfate, often a waste product of photoprocessing, is less bioavailable or toxic to aquatic organisms than is ionic silver. We conducted duplicate 48-h Ceriodaphnia dubia tests in reconstituted laboratory water using treatments of 92.7 nM Ag+ with various concentrations of thiosulfate. Expected Ag+ concentrations were generated for thiosulfate treatment levels using MINEQL + chemical equilibrium modeling. Ag+ concentrations in treatments were determined using a novel silicon-based sensor. Based on predicted Ag+ and published 48-h LC50 values for C. dubia, we did not expect to observe adverse effects. Yet, 100% mortality was observed at low thiosulfate treatments, whereas > 85% and > 95% survival was observed at higher thiosulfate treatment levels, respectively. Our results indicate that biotic responses match the sensor-based Ag+ concentrations. However, there is a discrepancy between these empirical results and responses expected to occur with Ag+ concentrations as predicted by MINEQL + chemical modeling. By correlating silicon sensor data with toxicity results obtained from our laboratory, our work clearly relates a specific chemical form (Ag+) to toxicity results.

Waterborne and sediment toxicity of fluoxetine to select organisms.

Ecological risk assessments of pharmaceuticals are currently difficult because little-to-no aquatic hazard and exposure information exists in the peer-reviewed literature for most therapeutics. Recently several studies have identified fluoxetine, a widely prescribed antidepressant, in municipal effluents. To evaluate the potential aquatic toxicity of fluoxetine, single species laboratory toxicity tests were performed to assess hazard to aquatic biota. Average LC(50) values for Ceriodaphnia dubia, Daphnia magna, and Pimephales promelas were 0.756 (234 microg/l), 2.65 (820 microg/l), and 2.28 microM (705 microg/l), respectively. Pseudokirchneriella subcapitata growth and C. dubia fecundity were decreased by 0.044 (14 microg/l) and 0.72 microM (223 microg/l) fluoxetine treatments, respectively. Oryias latipes survival was not affected by fluoxteine exposure up to a concentration of 28.9 microM (8.9 mg/l). An LC(50) of 15.2 mg/kg was estimated for Chironomus tentans. Hyalella azteca survival was not affected up to 43 mg/kg fluoxetine sediment exposure. Growth lowest observed effect concentrations for C. tentans and H. azteca were 1.3 and 5.6 mg/kg, respectively. Our findings indicate that lowest measured fluoxetine effect levels are an order of magnitude higher than highest reported municipal effluent concentrations.

Recommendations for the assessment of TNT toxicity in sediment.

Previous investigations of the ecotoxicity of TNT in spiked sediments noted the rapid degradation and disappearance of the toxicant, yet little is understood regarding the effects of this process on toxicity and subsequent derivation of toxicity reference values. We conducted environmental fate studies and 28-d sediment toxicity tests with benthic oligochaete worms (Tubifex tubifex) with sediments spiked at three different TNT concentrations (440, 1,409, and 4,403 nmol/g dry wt) aged for 1, 8, and 29 d. Because of rapid degradation of TNT, disappearance of degradation products, and partitioning to overlying water, only 25 to 40% of the added nitroaromatic mass balance was associated with sediment immediately after spiking. Lethal toxicity decreased with aging time and was best described by measured sediment nitroaromatic concentrations (sum of TNT and degradation products) at the beginning of exposure, with a median lethal concentration of nitroaromatic compounds of 184 nmol/g dry weight. To accurately describe the ephemeral exposure doses of TNT and its degradation products during toxicity tests with spiked sediments, we suggest that sediments should be aged at least 8 to 14 d after spiking, exposure should be based on measured sediment concentrations or chemical measures of availability, exchange of overlying water should be avoided or minimized, and short-term toxicity tests should be considered.

Laboratory and field responses to cadmium: an experimental study in effluent-dominated stream mesocosms.

Although select stream flows in the southwestern United States are dominated by effluent discharges, metal hazards have not been experimentally evaluated in effluent-dominated streams. Lotic mesocosms were designed to assess cadmium effects on multiple levels of biological organization, to determine relevance to regulatory criteria of standard laboratory toxicity tests, and to link laboratory tests to stream responses. Replicate streams were treated with 15 or 143 microg/L Cd during a 10-d study. Streams were sampled on days 0 and 10 for benthic macroinvertebrates, periphyton, and ecosystem metabolism. Concurrent Ceriodaphnia dubia and Pimephales promelas laboratory tests were performed with water from streams. Individual organism, population, and community response variables were affected by 143 microg/L Cd but not by the I5-microg/L treatment level. A biotic ligand model for cadmium predicted a 48-h median effective concentration (EC50) value of 280 microg/L Cd for C. dubia in these effluent-dominated streams: an EC50 value of 38.3 microg/L Cd was estimated for C. dubia in tests performed with reconstituted hard water. Our findings generally support use of the biotic ligand model for establishing site-specific, acute water quality criteria for cadmium. However, future effluent-dominated stream research is required to evaluate relationships between chronic cadmium exposure and organismal and community responses.

Tissue-specific bioconcentration of the synthetic steroid hormone medroxyprogesterone acetate in the common carp (Cyprinus carpio).

The steroid hormone medroxyprogesterone acetate (MPA), commonly used in oral and injectable contraceptives, has been detected in surface and wastewaters near urban and agricultural areas in several rivers of the world. The objectives of this study were to examine the accumulative potential and tissue distribution of MPA in fish. A freshwater species, the common carp (Cyprinus carpio), was exposed to 100 µg/L of MPA for a 7-day period followed by a depuration phase in which fish were maintained in dechlorinated tap water for an additional 7 days. Tissues (muscle, brain, plasma, and liver) were sampled during the uptake (days 1, 3, and 7) and depuration (day 14) phases of the experiment. Tissue-specific bioconcentration factors (BCF) ranged from 4.3 to 37.8 and uptake was greatest in the liver>brain>plasma and lowest in the muscle. From a regulatory standpoint, MPA shows little tendency to bioaccumulate in fish.

Nondestructive, minimal-disturbance, direct-burial solid-phase microextraction fiber technique for measuring TNT in sediment.

We explored a novel technique to deploy solid-phase microextraction (SPME) fibers to nondestructively measure the explosive compound 2,4,6-trinitrotoluene (TNT) and its nitroaromatic (NA) degradation products in laboratory sediment toxicity tests and field sediments in situ. SPME fibers within steel mesh envelopes were exposed statically via direct burial within sediment. Six fiber types (polymer coatings) were tested. Polyacrylate (PA) SPME fiber was sufficiently durable for this application, yielded the lowest detection limits, and exhibited a linear uptake relationship across toxicologically relevant sediment NA concentrations (100-2000 nmol/g dw (20-500 microg/g dw)). Temperature greatly influenced SPME absorption kinetics. Via evaluation of absorption at different temperatures, recommended sampling times needed to achieve steady-state equilibrium were 48 h for room temperatures (23-25 degrees C) and up to 7 d for cold (5 degrees C) temperatures. Although a comparison of TNT residues by SPMEs and TNT bioavailability and toxicity in sediments has not been completed, differences in SPME availability of TNT and its degradation products were found between two different TNT-spiked sediments. Our disposable SPME technique was slightly less expensive and as precise as the conventional extraction for total NAs and may prove to be a powerful exposure evaluation tool for assessing the ecological risk of these compounds.