Effects of Temperature and Salinity on Perfluorooctane Sulfonate (PFOS) Toxicity in Larval Estuarine Organisms.

Perfluorooctane sulfonate (PFOS) is a persistent contaminant that has been found globally within the environment. Key data gaps exist in the toxicity of PFOS to marine organisms, especially estuarine species that are crucial to the food web: fish, shrimp, and mollusks. This study developed toxicity thresholds for larval estuarine species, including grass shrimp (Palaemon pugio), sheepshead minnows (Cyprinodon variegatus), mysids (Americamysis bahia), and Eastern mud snails (Tritia obsoleta). Multiple abiotic stressors (salinity and temperature) were included as variables in testing the toxicity of PFOS. Acute 96 h toxicity testing under standard test conditions of 25 °C and 20 ppt seawater yielded LC50 values of 0.919 mg/L for C. variegatus, 1.375 mg/L for A. bahia, 1.559 mg/L for T. obsoleta, and 2.011 mg/L for P. pugio. The effects of increased temperature (32 °C) and decreased salinity (10 ppt) varied with test species. PFOS toxicity for the sheepshead minnows increased with temperature but was not altered by decreased salinity. For grass shrimp and mud snails, PFOS toxicity was greater under lower salinity. The combination of higher temperature and lower salinity was observed to lower the toxicity thresholds for all species. These data demonstrate that expanding toxicity testing to include a wider range of parameters will improve the environmental risk assessment of chemical contaminants, especially for species inhabiting dynamic estuarine ecosystems.

Mixture Effects of Per- and Polyfluoroalkyl Substances on Embryonic and Larval Sheepshead Minnows (Cyprinodon variegatus).

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous and persistent environmental contaminants originating from many everyday products. Perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) are two PFAS that are commonly found at high concentrations in aquatic environments. Both chemicals have previously been shown to be toxic to fish, as well as having complex and largely uncharacterized mixture effects. However, limited information is available on marine and estuarine species. In this study, embryonic and larval sheepshead minnows (Cyprinodon variegatus) were exposed to several PFAS mixtures to assess lethal and sublethal effects. PFOS alone was acutely toxic to larvae, with a 96 h LC50 of 1.97 mg/L (1.64-2.16). PFOS + PFOA resulted in a larval LC50 of 3.10 (2.62-3.79) mg/L, suggesting an antagonistic effect. These observations were supported by significant reductions in malondialdehyde (105% ± 3.25) and increases in reduced glutathione concentrations (43.8% ± 1.78) in PFOS + PFOA exposures compared to PFOS-only treatments, indicating reduced oxidative stress. While PFOA reduced PFOS-induced mortality (97.0% ± 3.03), perfluorohexanoic acid (PFHxA) and perfluorobutanoic acid (PFBA) did not. PFOS alone did not affect expression of peroxisome proliferator-activated receptor alpha (pparα) but significantly upregulated apolipoprotein A4 (apoa4) (112.4% ± 17.8), a downstream product of pparα, while none of the other individually tested PFAS affected apoa4 expression. These findings suggest that there are antagonistic interactions between PFOA and PFOS that may reduce mixture toxicity in larval sheepshead minnows through reduced oxidative stress. Elucidating mechanisms of toxicity and interactions between PFAS will aid environmental regulation and management of these ubiquitous pollutants.

Multi-stressor Effects of Ultraviolet Light, Temperature, and Salinity on Louisiana Sweet Crude Oil Toxicity in Larval Estuarine Organisms.

When oil is spilled into the environment its toxicity is affected by abiotic conditions. The cumulative and interactive stressors of chemical contaminants and environmental factors are especially relevant in estuaries where tidal fluctuations cause wide variability in salinity, temperature, and ultraviolet (UV) light penetration, which is an important modifying factor for polycyclic aromatic hydrocarbon (PAH) toxicity. Characterizing the interactions of multiple stressors on oil toxicity will improve prediction of environmental impacts under various spill scenarios. This study examined changes in crude oil toxicity with temperature, salinity, and UV light. Oil exposures included high-energy, water-accommodated fractions (HEWAFs) and thin oil sheens. Larval (24-48 h post hatch) estuarine species representing different trophic levels and habitats were evaluated. Mean 96 h LC50 values for oil prepared as a HEWAF and tested under standard conditions (20 ppt, 25 °C, No-UV) were 62.5 µg/L tPAH50 (mud snails), 198.5 µg/L (grass shrimp), and 774.5 µg/L (sheepshead minnows). Thin oil sheen 96 h LC50 values were 5.3 µg/L tPAH50 (mud snails), 14.7 µg/L (grass shrimp), and 22.0 µg/L (sheepshead minnows) under standard conditions. UV light significantly increased the toxicity of oil in all species tested. Oil toxicity also was greater under elevated temperature and lower salinity. Multi-stressor (oil combined with either increased temperature, decreased salinity, or both) LC50 values were reduced to 3 µg/L tPAH50 for HEWAFs and < 1.0 µg/L tPAH50 for thin oil sheens. Environmental conditions at the time of an oil spill will significantly influence oil toxicity and organismal response and should be taken into consideration in toxicity testing and oil spill damage assessments.

Uptake and Biological Effects of Perfluorooctane Sulfonate Exposure in the Adult Eastern Oyster Crassostrea virginica.

Perfluorooctane sulfonate (PFOS) is a legacy contaminant that has been detected globally within the environment and throughout numerous species, including humans. Despite an international ban on its use, this unique contaminant continues to persist in organisms and their surroundings due to PFOS's inability to breakdown into nontoxic forms resulting in bioaccumulation. In this study, we analyzed the effects of a technical mixture of PFOS (linear and branched isomers) in the adult Eastern oyster, Crassostrea virginica, at 2 days and 7 days exposure. Biomarker analysis (lysosomal destabilization, lipid peroxidation, and glutathione assays) in oyster tissue along with chemical analysis (liquid chromatography tandem mass spectrometry) of PFOS in oyster tissue and water samples revealed the oysters' ability to overcome exposures without significant damage to lipid membranes or the glutathione phase II enzyme system; however, significant cellular lysosomal damage was observed. The oysters were able to eliminate up to 96% of PFOS at 0.3 mg/L and 3 mg/L exposures when allowed to depurate for 2 days in clean seawater. Chemical analysis showed the linear isomer to be the prevailing fraction of the residual PFOS contained in oyster tissue. Results provide insight into possible detrimental cellular effects of PFOS exposure in addition to offering insight into contaminant persistence in oyster tissue.

Developmental and reproductive effects in grass shrimp (Palaemon pugio) following acute larval exposure to a thin oil sheen and ultraviolet light.

Many early stages of estuarine species congregate at the surface or in the upper mixing layer making them prone to UV light exposure and oil sheens. Laboratory testing was used to assess UV-oil sheen interactions with grass shrimp (Palaemon pugio). Newly hatched grass shrimp larvae were exposed to a 1-µm thick oil sheen for 24 h with or without an 8-h pulse of UV light. Grass shrimp were then transferred to clean seawater and non-UV conditions to measure development, growth, and reproductive fitness. Minimal toxicity was observed after the initial exposure but larval development was significantly delayed in shrimp exposed to the UV enhanced sheen. After reaching sexual maturity, shrimp were paired to evaluate effects on reproduction. Shrimp initially exposed to the UV enhanced sheen as larvae had a significant reduction in fecundity compared to controls. This demonstrates the importance of examining interactions between UV light and oil since negative effects to aquatic organisms may be underestimated if based on standard laboratory fluorescent lighting. Acute exposures of early life stages to thin oil sheens and UV light may lead to long-term impacts to individuals and ultimately to grass shrimp populations.

Toxicity comparison of the shoreline cleaners Accell Clean® and PES-51® in two life stages of the grass shrimp, Palaemonetes pugio.

Oil spills are a significant source of coastal pollution. Shoreline cleaners, used to remove oil from surfaces during spill response and remediation, may also act as toxins. Adult and larval grass shrimp, Palaemonetes pugio, were tested for lethal and sublethal impacts from two shoreline cleaners, Accell Clean SWA® and PES-51®, alone and in combination with crude oil using Chemically Enhanced Water Accommodated Fractions (CEWAFs). Median lethal toxicity values determined for the individual cleaners were similar. However, when tested in mixture with oil as CEWAFs, Accell Clean SWA resulted in greater hydrocarbon concentrations in the water column and greater toxicity than PES-51. Increased glutathione levels were observed for adult shrimp exposed to Accell Clean SWA, and glutathione was elevated in shrimp exposed to both CEWAFs. Larval shrimp development was delayed after exposure to both CEWAFs. These findings may have implications for managing and mitigating oil spills.

Effects of salinity on oil dispersant toxicity in the eastern mud snail, Ilyanassa obsoleta.

Chemical dispersants can be a beneficial method for breaking up oil slicks; however, their use in mitigation could pose potential toxic effects on the marine ecosystem. Dispersants may be transported to lower salinity habitats, where toxicity data for aquatic species have not been established. This study examined the effect of salinity on oil dispersant toxicity in the eastern mud snail, Ilyanassa obsoleta, using two dispersants authorized for oil spill response, Corexit® 9500A and Finasol® OSR 52. Median lethal toxicity values (LC50) and sublethal effects were examined at 10, 20, and 30 ppt salinity in adult and larval mud snails. Two biomarkers (lipid peroxidation and acetylcholinesterase) were used to measure sublethal effects. The 96-h static renewal LC50 values indicated significant differences in toxicity between dispersants and salinities. Larval snails were significantly more sensitive than adult snails to both dispersants, and both life stages were significantly more sensitive to Finasol than to Corexit. Larval snails were more sensitive to dispersants at lower salinity, but adult snails were more sensitive at higher salinities. Dispersants increased lipid peroxidation and decreased acetylcholinesterase activity. These results demonstrate that dispersant toxicity varies among compounds and organism life stages, and that physicochemical properties of the environment, such as salinity, can affect the potential toxicity to estuarine species.

Toxicity of oil and dispersant on the deep water gorgonian octocoral Swiftia exserta, with implications for the effects of the Deepwater Horizon oil spill.

Benthic surveys of mesophotic reefs in the Gulf of Mexico post Deepwater Horizon (DWH) showed that Swiftia exserta octocorals exhibited significantly more injury than in years before the spill. To determine the vulnerability of S. exserta to oil and dispersants, 96h toxicity assays of surrogate DWH oil water-accommodated fractions (WAF), Corexit® 9500 dispersant, and the combination of both (CEWAF) were conducted in the laboratory. Fragment mortality occurred within 48h for some fragments in the dispersant-alone and oil-dispersant treatments, while the WAF group remained relatively unaffected. The 96h LC50 values were 70.27mg/L for Corexit-alone and 41.04mg/L for Corexit in CEWAF. This study provides new information on octocoral sensitivity to toxins, and indicates that combinations of oil and dispersants are more toxic to octocorals than exposure to oil alone. These results have important implications for the assessment of effects of the DWH spill on deep-water organisms.

The ocean sampling day consortium.

Ocean Sampling Day was initiated by the EU-funded Micro B3 (Marine Microbial Biodiversity, Bioinformatics, Biotechnology) project to obtain a snapshot of the marine microbial biodiversity and function of the world's oceans. It is a simultaneous global mega-sequencing campaign aiming to generate the largest standardized microbial data set in a single day. This will be achievable only through the coordinated efforts of an Ocean Sampling Day Consortium, supportive partnerships and networks between sites. This commentary outlines the establishment, function and aims of the Consortium and describes our vision for a sustainable study of marine microbial communities and their embedded functional traits.

Individual and mixture effects of caffeine and sulfamethoxazole on the daggerblade grass shrimp Palaemonetes pugio following maternal exposure.

Pharmaceuticals and personal care products (PPCPs) such as caffeine and sulfamethoxazole have been detected in the estuarine environment. The present study characterized effects of a maternal exposure of these compounds on the development of the daggerblade grass shrimp Palaemonetes pugio from embryo to juvenile life stage. Ovigerous females were exposed to either caffeine (20 mg/L), sulfamethoxazole (60 mg/L), or a mixture of both (20 mg/L caffeine and 60 mg/L sulfamethoxazole). Embryos were then removed from the females and the effects of the PPCPs on hatching, metamorphosis, juvenile growth, and overall mortality were determined. No significant effect was observed on gravid female survival after 5 d of exposure to caffeine, sulfamethoxazole, or the mixture; however, development of the embryos on the female shrimp was delayed in the mixture. Caffeine and sulfamethoxazole in the mixture significantly reduced embryo survival. There was a significant effect of caffeine, sulfamethoxazole, and the mixture on embryo hatching time. Exposure to sulfamethoxazole alone significantly delayed larval metamorphosis. Exposure to caffeine and sulfamethoxazole separately led to significantly smaller length of juvenile shrimp. Maternal exposure to caffeine and sulfamethoxazole, individually and in mixture, resulted in negative effects on P. pugio offspring survival and development; however, the concentrations tested in the present study were well above maximum detected field concentrations. These results may be incorporated into PPCP risk assessments to protect sensitive estuarine ecosystems more effectively.

Comparative toxicity of pyrethroid insecticides to two estuarine crustacean species, Americamysis bahia and Palaemonetes pugio.

Pyrethroid insecticides are widely used on agricultural crops, as well as for nurseries, golf courses, urban structural and landscaping sites, residential home and garden pest control, and mosquito abatement. Evaluation of sensitive marine and estuarine species is essential for the development of toxicity testing and risk-assessment protocols. Two estuarine crustacean species, Americamysis bahia (mysids) and Palaemonetes pugio (grass shrimp), were tested with the commonly used pyrethroid compounds, lambda-cyhalothrin, permethrin, cypermethrin, deltamethrin, and phenothrin. Sensitivities of adult and larval grass shrimp and 7-day-old mysids were compared using standard 96-h LC50 bioassay protocols. Adult and larval grass shrimp were more sensitive than the mysids to all the pyrethroids tested. Larval grass shrimp were approximately 18-fold more sensitive to lambda-cyhalothrin than the mysids. Larval grass shrimp were similar in sensitivity to adult grass shrimp for cypermethrin, deltamethrin, and phenothrin, but larvae were approximately twice as sensitive to lambda-cyhalothrin and permethrin as adult shrimp. Acute toxicity to estuarine crustaceans occurred at low nanogram per liter concentrations of some pyrethroids, illustrating the need for careful regulation of the use of pyrethroid compounds in the coastal zone.

Influence of increasing temperature and salinity on herbicide toxicity in estuarine phytoplankton.

Ecological risk assessments are, in part, based on results of toxicity tests conducted under standard exposure conditions. Global climate change will have a wide range of effects on estuarine habitats, including potentially increasing water temperature and salinity, which may alter the risk assessment of estuarine pollutants. We examined the effects of increasing temperature and salinity on the toxicity of common herbicides (irgarol, diuron, atrazine, and ametryn) to the phytoplankton species Dunaliella tertiolecta. Static 96-h algal bioassays were conducted for each herbicide under four exposure scenarios: standard temperature and salinity (25°C, 20 ppt), standard temperature and elevated salinity (25°C, 40 ppt), elevated temperature and standard salinity (35°C, 20 ppt), and elevated temperature and elevated salinity (35°C, 40 ppt). The endpoints assessed were algal cell density at 96 h, growth rate, chlorophyll a content, lipid content, and starch content. Increasing exposure temperature reduced growth rate and 96-h cell density but increased the cellular chlorophyll and lipid concentrations of the control algae. Exposure condition did not alter starch content of control algae. Herbicides were found to decrease growth rate, 96 h cell density, and cellular chlorophyll and lipid concentrations, while starch concentrations increased with herbicide exposure. Herbicide effects under standard test conditions were then compared with those observed under elevated temperature and salinity. Herbicide effects on growth rate, cell density, and starch content were more pronounced under elevated salinity and temperature conditions. To encompass the natural variability in estuarine temperature and salinity, and to account for future changes in climate, toxicity tests should be conducted under a wider range of environmental conditions.

Comparative risk assessment of permethrin, chlorothalonil, and diuron to coastal aquatic species.

The precise application of risk assessment can lead to different conclusions about risk depending on how species are grouped in the assessment. We compared the use of different risk assessment methods for three different classes of pesticide, the herbicide diuron, the fungicide chlorothalonil, and the insecticide permethrin for marine and estuarine species. Permethrin was the most toxic pesticide to marine and estuarine crustaceans. Diuron was the most toxic pesticide to algae, and chlorothalonil was most toxic to early life stages of molluscs and other invertebrates. Toxicity data (96 h LC50/EC50 values) were analyzed using a probability distribution on the ranked toxicity values and 10th centile values were calculated based on different groups of species and for all species combined. Our results indicate that an assessment of risk based on smaller taxonomic groups can be informative, especially for pesticides of less specific modes of action such as chlorothalonil.

A long-term monitoring study of chlorophyll, microbial contaminants, and pesticides in a coastal residential stormwater pond and its adjacent tidal creek.

Stormwater ponds are commonly used in residential and commercial areas to control flooding. The accumulation of urban contaminants in stormwater ponds can lead to water-quality problems including nutrient enrichment, chemical contamination, and bacterial contamination. This study presents 5 years of monitoring data assessing water quality of a residential subdivision pond and adjacent tidal creek in coastal South Carolina, USA. The stormwater pond is eutrophic, as described by elevated concentrations of chlorophyll and phosphorus, and experiences periodic cyanobacterial blooms. A maximum monthly average chlorophyll concentration of 318.75 µg/L was measured in the stormwater pond and 227.63 µg/L in the tidal creek. Fecal coliform bacteria (FCB) levels were measured in both the pond and the tidal creek that exceeded health and safety standards for safe recreational use. A maximum monthly average FCB level of 1,247 CFU/100 mL was measured in the stormwater pond and 12,850 CFU/100 mL in the tidal creek. In addition, the presence of antibiotic resistant bacteria and pathogenic bacteria were detected. Low concentrations of herbicides (atrazine and 2,4-D: ), a fungicide (chlorothalonil), and insecticides (pyrethroids and imidacloprid) were measured. Seasonal trends were identified, with the winter months having the lowest concentrations of chlorophyll and FCB. Statistical differences between the stormwater pond and the tidal creek were also noted within seasons. The tidal creek had higher FCB levels than the stormwater pond in the spring and summer, whereas the stormwater pond had higher chlorophyll levels than the tidal creek in the summer and fall seasons. Chlorophyll and FCB levels in the stormwater pond were significantly correlated with monthly average temperature and total rainfall. Pesticide concentrations were also significantly correlated with temperature and rainfall. Pesticide concentrations in the stormwater pond were significantly correlated with pesticide concentrations in the adjacent tidal creek. Chlorophyll and FCB levels in the tidal creek, however, were not significantly correlated with levels in the pond. While stormwater ponds are beneficial in controlling flooding, they may pose environmental and human health risks due to biological and chemical contamination. Management to reduce residential runoff may improve water quality in coastal stormwater ponds and their adjacent estuarine ecosystems.

Effects of the synthetic pyrethroid insecticide, permethrin, on two estuarine fish species.

Limited toxicity data are available for estuarine and marine species and the widely used pyrethroid insecticide, permethrin. This study determined acute effects of permethrin on survival, lipid peroxidation, acetylcholinesterase activity, and splenocyte proliferation for two fish species found in South Carolina estuaries; juvenile red drum (Sciaenops ocellatus) and adult mummichog (Fundulus heteroclitus). Juvenile S. ocellatus were significantly more sensitive than adult F. heteroclitus to permethrin exposure, with a 96-h LC50 value of 8 µg/L determined for red drum compared to 23 µg/L for mummichog. Lipid peroxidation activity of the liver increased in permethrin-treated fish compared to control animals after 24 h and decreased after 96 h. Permethrin had no effect on acetylcholinesterase activity of the brain at the concentrations tested. Permethrin exposure significantly inhibited splenocyte proliferation, indicating an immunosuppressive effect. Most of the effects of permethrin on fish cellular stress enzymes and survival occurred at concentrations much higher than those typically measured in the environment. However, inhibition of splenocyte proliferation in juvenile red drum occurred at approximately twice that of measured permethrin concentrations in surface water. These findings may prove useful to the future management and regulation of pyrethroid insecticide use near estuarine habitats.

Toxicity of the mosquito control insecticide phenothrin to three life stages of the grass shrimp (Palaemonetes pugio).

Phenothrin is a synthetic pyrethroid used as a contact insecticide in mosquito control programs. This study compared the toxicity of phenothrin to adult, larval and embryonic grass shrimp (Palaemonetes pugio) and examined oxidative stress responses in adult and larval grass shrimp. The adult 24-h LC50 was 0.341 µg/L (95 % confidence intervals 0.282-0.412) and the 96-h LC50 was 0.161 µg/L (95 % CI 0.128-0.203 µg/L). The larval 24-h LC50 was 0.50 µg/L (95 % CI 0.441-0.568) and the 96-h LC50 was 0.154 µg/L (95 % CI 0.139-0.170 µg/L). In the presence of sediment, the 24-h LC50 was 6.30 µg/L (95 % CI 5.00-7.44 µg/L) for adults and 0.771 µg/L (95 % CI 0.630-0.944) for larvae. The sublethal biomarkers glutathione and lipid peroxidase (LPx) were examined after 96-h phenothrin exposure at five concentrations, and there were no statistically significant differences in these levels in adults or larvae compared to controls. There was a significant downward trend in larval LPx levels. This research confirms that phenothrin is highly toxic to grass shrimp and suggests that both adult and larval grass shrimp are appropriate life stages for risk assessments.

Toxicity of the insecticide etofenprox to three life stages of the grass shrimp, Palaemonetes pugio.

Opportunities for environmental contamination by the insecticide etofenprox are increasing as its uses expand from primarily indoor residential to rice cultivation and mosquito control. To provide toxicity data for sensitive saltwater species, effects of etofenprox were assessed using three life stages of the estuarine grass shrimp, Palaemonetes pugio. Adults, larvae, and embryos were tested in aqueous exposures, while adults and larval shrimp were also tested in the presence of sediment. In addition, sublethal cellular stress biomarkers, glutathione and lipid peroxidation, were examined. Larval shrimp was the most sensitive life stage, with 96-h median lethal concentration (LC(50)) of 0.89 microg/l, compared with 1.26 microg/l for adults and 100 microg/l for embryos. Presence of sediment significantly decreased toxicity of etofenprox to both adult and larval shrimp. Etofenprox exposure (100 micog/l) increased time to hatch in embryos. Lipid peroxidation levels were reduced in adult and larval shrimp after 96 h exposure to etofenprox, while no effect on glutathione was detected. The results of this study provide new information on the toxicity of etofenprox to estuarine invertebrates. These data may prove beneficial to the regulation of this pesticide and management of its uses in coastal areas.

Descriptive and mechanistic toxicity of conazole fungicides using the model test alga Dunaliella tertiolecta (Chlorophyceae).

Conazole fungicides are commonly used to prevent fungal growth on turf grass and agricultural crops. As many of these sites are adjacent to coastal waterways and estuaries, there exists the potential for nontarget effects of runoff on marine organisms. This study reports 96 h EC(50) values for four selected conazole fungicides (triadimefon = 5.98 mg/L; triadimenol = 5.51 mg/L; propiconazole = 2.33 mg/L; hexaconazole = 0.91 mg/L) to the model test alga Dunaliella tertiolecta. We further investigated possible mechanisms of toxicity by examining sublethal effects of exposure on cell morphology, osmoregulatory function, and lipid composition. These mechanistic studies revealed that conazole exposure does not inhibit synthesis of the cell's glycerol osmolyte, but does result in an overall increase in cellular volume and total lipid content. Both fungi and chlorophytes rely on ergosterol to maintain membrane structure and fluidity, and we provide evidence that the sterol-inhibiting conazoles may interfere with ergosterol biosynthesis in the cell membrane of Dunaliella. These findings suggest that green algae may be especially susceptible to nontarget effects of sterol-inhibiting fungicides in marine systems.

Temperature and salinity effects on the toxicity of common pesticides to the grass shrimp, Palaemonetes pugio.

This study investigated the effects of increased temperature and salinity, two potential impacts of global climate change, on the toxicity of two common pesticides to the estuarine grass shrimp, Palaemonetes pugio. Larval and adult grass shrimp were exposed to the fungicide chlorothalonil and the insecticide Scourge under standard toxicity test conditions, a 10 degrees C increase in temperature, a 10 ppt increase in salinity, and a combined increased temperature and salinity exposure. Toxicity of the fungicide chlorothalonil increased with temperature and salinity. Toxicity of the insecticide Scourge also increased with temperature; while increased salinity reduced Scourge toxicity, but only in adult shrimp. These findings suggest that changes in temperature and salinity may alter the toxicity of certain pesticides, and that the nature of the effect will depend on both the organism's life stage and the chemical contaminant. Standard toxicity bioassays may not be predictive of actual pesticide toxicity under variable environmental conditions, and testing under a wider range of exposure conditions could improve the accuracy of chemical risk assessments.

Lethal and sublethal toxicity of the antifoulant compound Irgarol 1051 to the mud snail Ilyanassa obsoleta.

Irgarol 1051 is an algistatic compound used in copper-based antifoulant paints. It is a widespread and persistent pollutant of the estuarine environment. Ilyanassa obsoleta, the Eastern mud snail, is a common intertidal gastropod that inhabits mud flats and salt marshes along the east coast of North America. It is an important inhabitant of the estuarine environment; contributing to nutrient regeneration and regulating microbial processes in the sediments. The toxicity of irgarol to estuarine gastropods has not been previously examined, although they have the potential to be exposed to antifoulants through both aqueous and sediment routes. The objectives of this study were to evaluate irgarol's effects on I. obsoleta survival, reproductive status (imposex occurrence and testosterone levels), chemoreceptive function, and cellular respiration (cytochrome-c oxidase activity). Irgarol was moderately toxic to I. obsoleta; adult aqueous 96-h LC(50) = 3.73 mg/L, larval aqueous 96-h LC(50) = 3.16 mg/L, and adult sediment 10-day LC(50) = 12.21 mg/kg. Larval snails were not significantly more sensitive to irgarol than adult snails. A chronic 45-day aqueous irgarol exposure (0.005-2.5 mg/L) did not induce imposex or affect free-testosterone levels. The 45-day chronic LC(50 )of 1.88 mg/L was significantly lower than the 96-h acute value. A 96-h acute aqueous irgarol exposure (0.375-1.5 mg/L) caused a decrease in normal response to chemosensory cues such as the presence of food or predators. There was a significant increase in cytochrome-c oxidase activity at 2.5 mg/L, which might indicate irgarol's disruption of the mitochondrial membrane and subsequently ATP synthesis. Although the toxicity values determined for I. obsoleta exceeded irgarol concentrations measured in surface waters, results from this toxicity assessment will provide valuable information to environmental resource managers faced with decisions regarding the use and regulation of antifoulant paints in the coastal zone.