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.

A Correlational Analysis of Phthalate Exposure and Thyroid Hormone Levels in Common Bottlenose Dolphins (Tursiops truncatus) from Sarasota Bay, Florida (2010-2019).

Phthalates are chemical esters used to enhance desirable properties of plastics, personal care, and cleaning products. Phthalates have shown ubiquitous environmental contamination due to their abundant use and propensity to leach from products to which they are added. Following exposure, phthalates are rapidly metabolized and excreted through urine. Common bottlenose dolphins (Tursiops truncatus) sampled from Sarasota Bay, Florida, have demonstrated prevalent di(2-ethylhexyl) phthalate (DEHP) exposure indicated by detectable urinary mono(2-ethylhexyl) phthalate (MEHP) concentrations. Widespread exposure is concerning due to evidence of endocrine disruption from human and laboratory studies. To better understand how phthalate exposure may impact dolphin health, correlations between relevant hormone levels and detectable urinary MEHP concentrations were examined. Hormone concentrations measured via blood serum samples included triiodothyronine (T3), total thyroxine (T4), and free thyroxine (FT4). Urinary MEHP concentrations were detected in 56% of sampled individuals (n = 50; mean = 8.13 ng/mL; s.d. = 15.99 ng/mL). Adult female and male FT4 was significantly correlated with urinary MEHP concentrations (adult female Kendall's tau = 0.36, p = 0.04; adult male Kendall's tau = 0.42, p = 0.02). Evidence from this study suggests DEHP exposure may be impacting thyroid hormone homeostasis. Cumulative effects of other stressors and resultant endocrine impacts are unknown. Further research is warranted to understand potential health implications associated with this relationship.

Assessment of the efficacy of six field cleaning protocols for hydrocarbon quantification.

The defensibility of field sampling data collected in support of natural resource damage assessments and other environmental investigations depends on rigorous quality assurance and control both in the field and laboratory. One important step in field procedures is the cleaning of sampling equipment between samples to minimize the carryover of contaminants. Large-scale sampling efforts during the Deepwater Horizon oil spill event have highlighted the importance of understanding how multiple equipment cleaning protocols affect interstation cross-contamination and the resulting chemical data quality. In this study, six field cleaning techniques were tested on metal sampling equipment using two different sediment types spiked with crude oil in order to understand their relative and absolute effectiveness in reducing chemical carryover. The complexity of the cleaning protocols ranged from a simple water and scrub brush application to protocols that included soap and/or solvent. In this study, percent residual hydrocarbon transfer, relative to total loading in sediments, never exceeded 0.032%. The least labor-intensive protocol, water and scrub brush application, had the highest potential for hydrocarbon transfer (0.011-0.032%). Statistical differences were observed among treatments, and it was found that protocols containing a solvent step were more effective than protocols without solvents. Depending on the data quality objectives, the differences may not be meaningful, and choosing a cleaning technique should be governed by health, safety, and environmental factors. The residual hydrocarbons measured after equipment cleanings for all techniques in this study were negligible when compared with other variables that occur during routine sampling and laboratory activities.

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.

Sentinels of synthetics - a comparison of phthalate exposure between common bottlenose dolphins (Tursiops truncatus) and human reference populations.

Phthalates are chemical esters used as additives in common consumer goods, such as plastics, household cleaners, and personal care products. Phthalates are not chemically bound to the items to which they are added and can easily leach into the surrounding environment. Anthropogenic drivers, such as coastal plastic pollution and wastewater runoff, increase the exposure potential for coastal marine fauna. Phthalate exposure in free-ranging bottlenose dolphins has been the focus of recent study, with indications of heightened exposure to certain phthalate compounds. The objective of this study was to compare urinary phthalate metabolite concentrations among bottlenose dolphins (Tursiops truncatus) sampled in Sarasota Bay, FL, to levels reported in human samples collected as part of the Centers for Disease Control and Prevention's (CDC) National Health and Nutrition Examination Survey (NHANES). Monoethyl phthalate (MEP) and mono-(2-ethylhexyl) phthalate (MEHP) were the most prevalent metabolites detected in dolphin urine (n = 51; MEP = 29.41%; MEHP = 54.90%). The geometric mean (GM) concentration of MEP was significantly lower for dolphins (GM = 4.51 ng/mL; 95% CI: 2.77-7.34 ng/mL) compared to humans (p<0.05), while dolphin concentrations of MEHP (GM = 4.57 ng/mL; 95% CI: 2.37-8.80 ng/mL) were significantly higher than levels reported in NHANES (p<0.05). Health impacts to bottlenose dolphins resulting from elevated exposure to the MEHP parent compound (diethyl-2-ethylhexyl phthalate, DEHP) are currently unknown. However, given the evidence of endocrine disruption, reproductive impairment, and abnormal development in humans, pursuing investigations of potential health effects in exposed bottlenose dolphins would be warranted.

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.