Sean M. Richards

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.

Effects of 25 pharmaceutical compounds to Lemna gibba using a seven-day static-renewal test

Antibiotics are known to have antichloroplastic properties, but their effects on aquatic higher plants are virtually unknown. In order to address this issue, 25 pharmaceuticals, including 22 antibiotics, were assessed for phytotoxicity to the aquatic higher plant Lemna gibba. A 7-d static-renewal test was used, and plants were treated with 0, 10, 30, 100, 300, and 1,000 microg/L of pharmaceutical-containing growth media. Phytotoxicity was assessed using multiple growth and biochemical endpoints. Effective concentration (EC)50, EC25, and EC10 values as well as tests for significant differences between treatments and controls lowest-observed-effect concentration (LOECs) were calculated for each endpoint. Twelve different classes of antibiotics were assessed; however, only members of the fluoroquinolone, sulfonamide, and tetracycline classes of antibiotics displayed significant phytotoxicity. The most toxic members of each of these classes tested were lomefloxacin, sulfamethoxazole, and chlortetracycline, with wet weight EC25 values of 38, 37, and 114 microg/L, respectively. Injury symptoms were comparatively uniform and consistent among chemical classes while degree of phytotoxicity varied considerably. Both of these criteria varied markedly between classes. Wet mass was consistently the most sensitive endpoint above 100 microg/L; conversely, frond number was the most sensitive below 100 microg/L. Pigment endpoints were significantly less sensitive than growth endpoints.

Aquatic persistence of eight pharmaceuticals in a microcosm study

The persistence of eight pharmaceuticals from multiple classes was studied in aquatic outdoor field microcosms. A method was developed for the determination of a mixture of acetaminophen, atorvastatin, caffeine, carbamazepine, levofloxacin, sertraline, sulfamethoxazole, and trimethoprim at microg/L levels from surface water of the microcosms using solid phase extraction and high-performance liquid chromatography-ultraviolet (HPLC-UV) and liquid chromatography tandem mass spectrometry (LC-MS-MS). Half-lives in the field ranged from 1.5 to 82 d. Laboratory persistence tests were performed to determine the relative importance of possible loss processes in the microcosms over the course of the study. Results from dark control experiments suggest hydrolysis was not important in the loss of the compounds. No significant differences were observed between measured half-lives of the pharmaceuticals in sunlight-exposed pond water and autoclaved pond water, which suggests photodegradation was important in limiting their persistence, and biodegradation was not an important loss process in surface water over the duration of the study. Observed photoproducts of several of the pharmaceuticals remained photoreactive, which led to further degradation in irradiated surface waters.

Effects of pharmaceutical mixtures in aquatic microcosms.

Pharmaceuticals have a wide range of biological properties and are released into the environment in relatively large amounts, yet little information is available regarding their effects or potential ecological risks. We exposed outdoor aquatic microcosms to combinations of ibuprofen (a nonsteroidal anti-inflammatory drug), fluoxetine (a selective serotonin reuptake inhibitor), and ciprofloxacin (a DNA gyrase-inhibiting antibiotic) at concentrations of 6, 10, and 10 microg/L, respectively (low treatment [LT]); 60, 100, and 100 microg/L, respectively (medium treatment [MT]); and 600, 1,000, and 1,000 microg/L, respectively (high treatment [HT]). We maintained these concentrations for 35 d. Few responses were observed in the LT; however, effects were observed in the MT and HT. Fish mortality occurred in the MT (<35 d) and in the HT (<4 d). Phytoplankton increased in abundance and decreased in diversity (number of taxa) in the HT, with consistent trends being observed in the MT and LT. Zooplankton also showed increased abundance and decreases in diversity in the HT, with consistent trends being observed in the MT. Multivariate analyses for zooplankton and phytoplankton suggested interactions between these communities. Lemna gibba and Myriophyllum spp. showed mortality in the HT; growth of L. gibba was also reduced in the MT. Bacterial abundance did not change in the HT. All responses were observed at concentrations well below the equivalent pharmacologically active concentrations in mammals. Although the present data do not suggest that ibuprofen, fluoxetine, and ciprofloxacin are individually causing adverse effects in surface-water environments, questions remain about additive responses from mixtures.

Rapid liquid chromatography–tandem mass spectrometry method for the determination of a broad mixture of pharmaceuticals in surface water

Herein, a new method for the detection of 13 different pharmaceuticals and one metabolite in surface water at low ng/L levels is described. The method utilizes ultra performance liquid chromatography-tandem mass spectrometry and a solid-phase extraction sample preparation. Mean method detection limits were low (4.10 ng/L) and overall solid-phase extraction recovery and reproducibility was adequate (mean recovery, 77.9%; mean RSD, 7.3%). The method allows for quick run times and minimal solvent use as compared with other previously reported high performance liquid chromatography-based methods. Application of this method for the detection of pharmaceuticals in Tennessee River surface water determined that caffeine, sulfamethoxazole, and carbamazepine were frequently detected (100% of samples). Trimethoprim was moderately detected (30% of samples); acetaminophen, atorvastatin, and lovastatin were infrequently detected (10% of samples); and ciprofloxacin, diltiazem, fluoxetine, levofloxacin, norfluoxetine, ranitidine, and sertraline were not detected. This study reports the first detection of lovastatin in surface water.

Spatial and temporal analysis of pharmaceutical concentrations in the upper Tennessee River basin

The behavior of pharmaceutical compounds in aquatic ecosystems is not well defined. In order to determine spatial and temporal variations in concentrations of pharmaceuticals in the Tennessee River, water samples were collected from multiple points along the river and at the inflow of major tributaries. Sampling structure was designed to investigate trends between surface and subsurface samples, seasonal trends (winter, spring, summer, and fall), the direct influence of sewage treatment plants (upstream versus downstream), and the effect of downstream distance on pharmaceutical concentrations. All samples were quantified via solid phase extraction followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This method yielded reproducible quantitation at low parts per trillion (ng L(-1)) levels for all 14 analytes (acetaminophen, atorvastatin, caffeine, carbamazepine, ciprofloxacin, diltiazem, fluoxetine, levofloxacin, lovastatin, norfluoxetine, ranitidine, sertraline, sulfamethoxazole, and trimethoprim). Correlation analyses (depth, distance) and repeated-measures ANOVAs (season, sewage treatment plant proximity) were used to determine statistically significant trends for frequently detected pharmaceuticals (caffeine, carbamazepine, sulfamethoxazole). Caffeine and sulfamethoxazole were found to vary by season in subsurface samples; spring exhibited the highest concentrations. Carbamazepine varied in proximity to sewage treatment plant outfall with subsurface samples yielding greater concentrations downstream than upstream. In addition, individual pharmaceuticals displayed positive correlation between surface and subsurface samples and negative correlation with downstream distance from the headwaters.

Placental 11β-Hydroxysteroid dehydrogenase type 2 expression: Correlations with birth weight and placental metal concentrations.

INTRODUCTION Infants born below 2500 g are classified as low birth weight. Excess in utero exposure to cortisol has been linked to restricted fetal growth. Placental production of 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) inactivates cortisol before passage into the fetus. The present study tested the hypothesis that placental 11β-HSD2 expression is positively correlated with an individualized birth weight centile and raw birth weight, and examines the relationship between metal concentrations in placental tissue and 11β-HSD2 expression. METHODS Placentae from 191 births were collected and samples preserved to maintain mRNA profile. Placental 11β-HSD2 expression was measured via qRT-PCR. Addition samples were collected from placental tissues and uniformly dried in order to quantify 18 metals via ICP-MS (n = 160). RESULTS A significant, positive correlation between 11β-HSD2 expression and individualized birth weight centile (p = 0.0321) and birth weight (p = 0.0243) was found. Additionally, maternal age and gestational age were positivity correlated with each other (p = 0.0321). Birth weight was significantly different with race, marital status, education and maternal tobacco use. Four metals (Co, Mn, Ni, Zn) demonstrated significant positive correlations (p < 0.05) with 11β-HSD2 expression. Sex specific differences were found; Co, Cu, Fe, Zn, and Ni were positively correlated with 11β-HSD2 expression in males only, no significant correlations were found in the female only sample. CONCLUSION These data indicate that the growth potential of a fetus is related to the 11β-HSD2 expression in the placenta, and that 11β-HSD2 expression is related to the trace metals status of the mother.

Effects of stream acidification on lotic salamander assemblages in a coal-mined watershed in the Cumberland Plateau

We studied the effects of acid mine drainage (AMD) from abandoned coal mines on lotic salamanders and environmental conditions in the upper watershed (Cumberland Plateau) of North Chickamauga Creek (NCC; Tennessee River drainage) in southeastern Tennessee, USA, from 1996–97. Study sites (2nd- or 3rd-order reaches) were sampled in an AMD-influenced section (five sites) and in two reference streams (two minimally disturbed sites). A total of 212 plethodontids (premetamorphic larvae) representing four species were collected by kicknetting in riffles (n = 99) and electrofishing in mixed habitats (n = 113). The dusky salamander (Desmognathus fuscus) was the most abundant species in both AMD and reference reaches (> 80 – 90% of total catches), successively followed by the southern two-lined salamander (Eurycea cirrigera), spring salamander (Gyrinophilus porphyriticus), and red salamander (Pseudotriton ruber). Mining-influenced reaches were characterized by acidic flows (mean pH = 3.8–5.6), zero to low alkalinity, and elevated conductivity, sulfate, hardness, aluminum, and manganese, as well as very low abundances of salamanders. Reference reaches were slightly acidic to circumneutral (mean pH = 6.0–6.9) with low to moderate alkalinity, low levels of conductivity, hardness, sulfate, and metals, and high salamander abundances. Our findings document the impact of acid/metal pollution from past coal mining activities on lotic salamanders in a Cumberland Plateau stream.

Is ambient chitobiase activity a monitoring tool for impacts on secondary production in lotic systems

Chitobiase is one of two enzymes responsible for cleaving the chitin polymers that form arthropod exoskeletons and has been found to be predictive of secondary production for marine and freshwater zooplankton. Since chitobiase is re- leased into water during molting by aquatic arthropods, it is possible that ambient chitobiase activity may be a useful proxy of secondary production in aquatic systems. In the present study, surface water samples from the upper Tennessee River basin, USA, were collected and analyzed for chitobiase to determine how enzyme levels vary with season, water depth, and proximity to sewage treatment outfalls. Additionally, chitobiase activity was correlated with three pharmaceuti- cal compounds to assess if changes in chitobiase activity were associated with contaminants derived from sewage treat- ment plants. Chitobiase activity was significantly (p < 0.05) elevated in the fall relative to other seasons. Further, chitobiase activity displayed significant positive correlation (p < 0.05) between surface and subsurface samples and was negatively correlated (p < 0.05) with pharmaceutical concentrations in summer samples. These results indicate the potential for chitobiase as an assessment tool in estimating secondary production and as an ecotoxicological indictor.

Environmental toxicant exposure during pregnancy.

This literature review is meant to serve as a brief reference for acute and chronic poisonings in pregnant women, specifically involving environmental toxicants commonly present in the home or workplace. These scenarios are familiar to primary care providers but cause great confusion for practitioners and anxiety in the pregnant patient. Herein, we review metals and metalloids, organic solvents, disinfectant byproducts, pesticides, plasticizers, and multiple air pollutants. Reviews of specific studies involving these toxicants are provided to assist practitioners in providing information to patients regarding potential sources, mechanism of action, current laboratory and epidemiological studies, and possible treatments. Literature-based associations with specific toxicants and various pregnancy outcomes are also outlined. Finally, a contact list of important federal and state toxicology support services is provided. Target Audience: Obstetricians & Gynecologists Learning objectives: After completing this CME activity, physicians should be better able to assess both acute and chronic consequences of various environmental toxic exposures in pregnancy; to evaluate possible pregnancy related specific events surrounding environmental pollutants; and to identify common exposure routes and implement therapeutic interventions where appropriate.