How weathering might intensify the toxicity of spilled crude oil in marine environments.

Crude oils are highly complex mixtures containing many toxic compounds for organisms. While their level of toxicity in a marine environment depends on many parameters, one of the main factors is their composition. After oil spills, their compositions are significantly changed, so it changes the toxicity. In this study, different weathering processes such as evaporation, photooxidation, and biodegradation were applied to crude oil to understand how composition changed over time and how this affects its toxicity on phytoplankton. In laboratory settings, three distinct water-accommodated fraction samples of crude oil were prepared, unweathered, evaporated, and weathered and were exposed to phytoplankton communities at different dilution levels. After 3 days, evaporation reduced the crude oil concentration by 47%, and the concentration of the crude oil affected by photooxidation, biodegradation, and evaporation reduced by 81%. This study also showed that even though the weathering reduced the overall amount of crude oil substantially, its toxicity increased significantly. In the microcosm experiments, 7-day EC50 values of the unweathered oil, the evaporated oil and the weathered oil were 49.07, 21.09, and 7.16 µg/L, respectively. Different processes altered the crude oil composition, and weathered crude oil ended up with a higher fraction of high molecular weight (HMW) polycyclic aromatic hydrocarbons (PAHs). A promising relation between the increasing toxicity and HMW PAH fraction indicates that increasing the fraction of HMW PAHs might be one of the main reasons for the weathering process to cause higher crude oil toxicity. These results could be used as a diagnostic tool to estimate the extent of weathering and toxicity of crude oil after spills.

Marine phytoplankton responses to oil and dispersant exposures: Knowledge gained since the Deepwater Horizon oil spill.

The Deepwater Horizon oil spill of 2010 brought the ecology and health of the Gulf of Mexico to the forefront of the public's and scientific community's attention. Not only did we need a better understanding of how this oil spill impacted the Gulf of Mexico ecosystem, but we also needed to apply this knowledge to help assess impacts from perturbations in the region and guide future response actions. Phytoplankton represent the base of the food web in oceanic systems. As such, alterations of the phytoplankton community propagate to upper trophic levels. This review brings together new insights into the influence of oil and dispersant on phytoplankton. We bring together laboratory, mesocosm and field experiments, including insights into novel observations of harmful algal bloom (HAB) forming species and zooplankton as well as bacteria-phytoplankton interactions. We finish by addressing knowledge gaps and highlighting key topics for research in novel areas.

Trimethyl 4,4',4''-(ethene-1,1,2-tri-yl)tribenzoate.

The title compound, C26H22O6, is formed as the major product from the reaction between syn-1,2-bis-(pinacolatoboron)-1,2-bis-(4-methyl-carb-oxy-phen-yl)ethene and excess methyl 4-iodo-benzoate in basic DMSO using a palladium catalyst at 80°C via Suzuki coupling followed by protodeboronation. Crystals were grown by slow evaporation of a hexa-nes solution at room temperature.

Degradation of Bisphenol A in Natural and Artificial Marine and Freshwaters in Turkey.

Bisphenol A (BPA), one of the important synthetic chemicals, has been produced at high volumes since the 1960s. These chemicals are commonly detected in the marine and freshwater environments; however, their transformation in aquatic environments depends on many parameters. This study aims to investigate the degradation of BPA in marine and freshwaters under different conditions in terms of microbial degradation, photodegradation, and temperature effect. The results showed that BPA content in samples prepared from the artificial waters did not change significantly in 150 days. BPA concentrations in natural river water started to degrade after day 50, and the degradation rate was faster for the samples at 25°C than ones at 4°C. In natural seawater samples, there was no degradation detected in 150 days at 4°C and 25°C. However, samples prepared in natural seawater, kept outside, and exposed to over 40°C temperature showed degradation after day 50. A treatment exposed to the sunlight showed a higher degradation rate, indicating the additive/synergistic role of the photodegradation. Our study suggests that high temperatures ( > 25°C) are required for BPA degradation in seawater. River water is more potent than seawater for BPA degradation. It is suggested that BPA contamination in a marine environment could be more persistent than in a freshwater environment.

Temporal and Spatial Distributions of Bisphenol A in Marine and Freshwaters in Turkey.

Bisphenol A (BPA), a chemical component used in the manufacture of plastics, is commonly introduced to and detected in aquatic environments. This is the first study conducted to understand the distribution of BPA in the marine and freshwaters of Turkey. The purpose of this study is to report BPA concentrations measured from a time-series conducted in coastal waters of Erdemli and regional rivers located in the northeastern Mediterranean region. Furthermore, seawater samples obtained from other Turkish coastal areas-The Black Sea, Bosphorus, Sea of Marmara, and the Mediterranean Sea-also were investigated to gain a better understanding of regional and seasonal variations of BPA concentrations in Turkish Seas. Whilst spatial variation in BPA concentrations was very low, temporal variation was found to be high. It has been shown that BPA can reach the deep sea environment (> 500 m depth). This study indicated that BPA contamination has reached serious levels at another location in the world.

Assessment of trophic status of the northeastern Mediterranean coastal waters: eutrophication classification tools revisited.

The Eastern Mediterranean and its Cilician Basin offshore waters have oligotrophic features with low nutrient concentrations, low primary production, and high water transparency. However, the wide shelf area of the Cilician Basin is subject to contaminated river inflows with enhanced nutrient loads and direct discharges of urban wastewaters of southern Turkey, leading to develop local eutrophic/mesotrophic conditions in the inner sites of Mersin and Iskenderun Bays on the Cilician Basin. For the assessment of changing trophic status of the coastal and the bay water bodies under anthropogenic pressures since the 1980s, five extensive field studies were performed in summer and winter periods of 2014, 2015, and 2016. Physical and eutrophication-related biochemical parameters (salinity, temperature, Secchi Disk Depth, nutrients, dissolved oxygen, chlorophyll-a) were measured at 65 stations in different water bodies occupying the Northeastern (NE) Mediterranean coastal, offshore areas and bays. The collected data sets were used in scaling the trophic status of the visited water bodies of NE Mediterranean coastal, offshore areas and semi-enclosed bays, using novel classification tools of Trophic Index (TRIX), Eutrophication Index (E.I.), chl-a, and HELCOM Eutrophication Assessment Tool (HEAT), developed by different experts for highly productive seas. These tools, which can successfully classify highly productive coastal water masses under human pressures, and their sensitivities have been tested for scaling of the current trophic status of the NE Mediterranean coastal water bodies being subject to human pressures. The scaling results of classical TRIX, E.I., and chl-a indices in the NE Mediterranean water masses are not sensitive enough to differentiate mesotrophic and eutrophic water bodies because these indices principally assume to have higher concentrations of eutrophication-related parameters in the least effected (reference) water bodies. The HEAT tool, which uses a site-specific "reference value" for each eutrophication-indicator, has allowed us to produce more reliable and sensitive scaling of the current trophic status of the NE Mediterranean shelf areas, even though we used only the "reference values" derived from the composite data sets. The results of the indices were compared with the HEAT tool and the actual status was assessed from observations, indicating revision requirements of the multi-metric classification tools. For this goal, scales of natural (oligotrophic) and anthropogenic (eutrophic) levels of eutrophication indicators should be determined at a sub-basin scale using long-term site-specific observations in the NE Mediterranean. The revised scale ranges of TRIX for oligotrophic, mesotrophic, and eutrophic water bodies of Mersin Bay are in line with ranges of TRIX classification tool proposed for Aegean Sea waters, which can be used to assess trophic status of the entire Eastern Mediterranean and Aegean coastal seas (surface salinity > 37.5) having oligotrophic properties in the offshore waters.

Temperature Effects Explain Continental Scale Distribution of Cyanobacterial Toxins.

Insight into how environmental change determines the production and distribution of cyanobacterial toxins is necessary for risk assessment. Management guidelines currently focus on hepatotoxins (microcystins). Increasing attention is given to other classes, such as neurotoxins (e.g., anatoxin-a) and cytotoxins (e.g., cylindrospermopsin) due to their potency. Most studies examine the relationship between individual toxin variants and environmental factors, such as nutrients, temperature and light. In summer 2015, we collected samples across Europe to investigate the effect of nutrient and temperature gradients on the variability of toxin production at a continental scale. Direct and indirect effects of temperature were the main drivers of the spatial distribution in the toxins produced by the cyanobacterial community, the toxin concentrations and toxin quota. Generalized linear models showed that a Toxin Diversity Index (TDI) increased with latitude, while it decreased with water stability. Increases in TDI were explained through a significant increase in toxin variants such as MC-YR, anatoxin and cylindrospermopsin, accompanied by a decreasing presence of MC-LR. While global warming continues, the direct and indirect effects of increased lake temperatures will drive changes in the distribution of cyanobacterial toxins in Europe, potentially promoting selection of a few highly toxic species or strains.

Induction of reactive oxygen species in marine phytoplankton under crude oil exposure.

Exposure of phytoplankton to the water-accommodated fraction of crude oil can elicit a number of stress responses, but the mechanisms that drive these responses are unclear. South Louisiana crude oil was selected to investigate its effects on population growth, chlorophyll a (Chl a) content, antioxidative defense, and lipid peroxidation, for the marine diatom, Ditylum brightwellii, and the dinoflagellate, Heterocapsa triquetra, in laboratory-based microcosm experiments. The transcript levels of several possible stress-responsive genes in D. brightwellii were also measured. The microalgae were exposed to crude oil for up to 96 h, and Chl a content, superoxide dismutase (SOD), the glutathione pool (GSH and GSSG), and lipid peroxidation content were analyzed. The cell growth of both phytoplankton species was inhibited with increasing crude oil concentrations. Crude oil exposure did not affect Chl a content significantly in cells. SOD activities showed similar responses in both species, being enhanced at 4- and 8-mg/L crude oil exposure. Only H. triquetra demonstrated enhanced activity in GSSG pool and lipid peroxidation at 8-mg/L crude oil exposure, suggesting that phytoplankton species have distinct physiological responses and tolerance levels to crude oil exposure. This study indicated the activation of reactive oxygen species (ROS) in phytoplankton under crude oil exposure; however, the progressive damage in cells is still unknown. Thus, ROS-related damage in nucleic acid, lipids, proteins, and DNA, due to crude oil exposure could be a worthwhile subject of study to better understand crude oil toxicity at the base of the food web.

Responses of sympatric Karenia brevis, Prorocentrum minimum, and Heterosigma akashiwo to the exposure of crude oil.

Impacts of the Deepwater Horizon oil spill on phytoplankton, particularly, the tolerability and changes to the toxin profiles of harmful toxic algal species remain unknown. The degree to which oil-affected sympatric Karenia brevis, Prorocentrum minimum, and Heterosigma akashiwo, all of which are ecologically important species in the Gulf of Mexico, was investigated. Comparison of their tolerability to that of non-toxic species showed that the toxin-production potential of harmful species does not provide a selective advantage. Investigated toxin profiles for K. brevis and P. minimum demonstrated an increase in toxin productivity at the lowest crude oil concentration (0.66 mg L(-1)) tested in this study. Higher crude oil concentrations led to significant growth inhibition and a decrease in toxin production. Findings from this study could assist in the assessment of shellfish bed closures due to high risk of increased toxin potential of these phytoplankton species, especially during times of stressed conditions.

Relative phytoplankton growth responses to physically and chemically dispersed South Louisiana sweet crude oil.

We conducted controlled laboratory exposure experiments to assess the toxic effects of water-accommodated fractions (WAFs) of South Louisiana sweet crude oil on five phytoplankton species isolated from the Gulf of Mexico. Experiments were conducted with individual and combinations of the five phytoplankton species to determine growth inhibitions to eight total petroleum hydrocarbon (TPH) equivalent concentrations ranging from 461 to 7,205 ppb. The composition and concentration of crude oil were altered by physical and chemical processes and used to help evaluate crude oil toxicity. The impact of crude oil exposure on phytoplankton growth varied with the concentration of crude oil, species of microalgae, and their community composition. At a concentration of TPH < 1,200 ppb, dinoflagellate species showed significantly better tolerance, while diatom species showed a higher tolerance to crude oil at higher concentrations of TPH. For both groups, the larger species were more tolerant to crude oil than smaller ones. The toxicity potential of crude oil seems to be strongly influenced by the concentration of polycyclic aromatic hydrocarbons (PAHs). The addition of the dispersant, Corexit® EC9500A, increased the amount of crude oil up to 50-fold in the water column, while the physical enhancement (vigorous mixing of water column) did not significantly increase the amount of TPH concentration in the water column. The species response to crude oil was also examined in the five-species community. Each phytoplankton species showed considerably less tolerance to crude oil in the five-species community compared to their individual responses. This study provides baseline information about individual phytoplankton responses to crude oil and dispersed crude oil for subsequent research efforts seeking to understand the impacts of oil on the phytoplankton in the bigger picture.

Distinct responses of Gulf of Mexico phytoplankton communities to crude oil and the dispersant corexit(®) Ec9500A under different nutrient regimes.

This study examines the potential effects of exposure to South Louisiana sweet crude oil (LSC), Corexit(®) EC9500A, and dispersed oil on enclosed phytoplankton communities under different nutrient regimes. Three distinct microcosm experiments were conducted for 10 days to assess changes to the structure of natural communities from the Gulf of Mexico as quantified by temporal changes in the biomasses of different phytoplankton groups. Concentration of NO3, Si and PO4 were 0.83, 0.99 and 0.09 µM for the unenriched treatments and 14.07, 13.01 and 0.94 µM for the enriched treatments, respectively. Overall, the contaminants LSC and Corexit(®) EC9500A led to a decrease in the number of sensitive species and an increase in more resistant species. Phytoplankton communities showed more sensitivity to LSC under nutrient-limited conditions. The addition of nutrients to initially nutrient-limited treatments lessened the inhibitory effect of LSC in the short term. Centric diatoms benefited most from this enrichment, but pennate diatoms demonstrated considerably greater tolerance to crude oil at low crude oil concentrations in nutrient-enriched treatments. Dinoflagellates showed relatively higher tolerance in nutrient-limited treatments and high crude oil concentrations. Corexit(®) EC9500A inputs significantly increased the toxicity of crude oil. Corexit(®) EC9500A alone had a highly inhibitory effect at 63 ppm on phytoplankton communities. This study highlights the fact that different nutrient regimes play a major role in determining the shifts of the phytoplankton community in response to exposure to different concentrations of crude oil and dispersant. Determination of the functional equivalence of shifted phytoplankton groups could complement our research and allow for more pertinent extrapolation to real world conditions.

Can crude oil toxicity on phytoplankton be predicted based on toxicity data on benzo(a)pyrene and naphthalene?

Polycyclic aromatic hydrocarbons (PAHs), which are major components of crude oil, are responsible in large part for the toxicity of crude oil to phytoplankton. This study addressed the following question. Can reliable predictions of the aquatic toxicity of crude oil, a multi-component mixture, be described from toxicity data on individual PAH compounds? Naphthalene, the most abundant PAH compound, and benzo(a)pyrene, a highly toxic PAH compound, were selected as model compounds to quantify toxicity of crude oil on two phytoplankton species, Ditylum brightwellii and Heterocapsa triquetra, by analyzing the effects of different concentrations of these PAHs on growth rate. EC50 values suggested that the diatom D. brightwellii was more vulnerable to both toxicants than the dinoflagellate H. triquetra. However, a previous study, which investigated the impact of crude oil on the same two species, had opposite results. The differences in response from these phytoplankton species to naphthalene and benzo(a)pyrene toxicity compared to their response to crude oil suggest that they may not be solely used as surrogates to assess crude oil toxicity on phytoplankton.

Ultra-rapid absorption of recombinant human insulin induced by zinc chelation and surface charge masking.

BACKGROUND: In order to enhance the absorption of insulin following subcutaneous injection, excipients were selected to hasten the dissociation rate of insulin hexamers and reduce their tendency to reassociate postinjection. A novel formulation of recombinant human insulin containing citrate and disodium ethylenediaminetetraacetic acid (EDTA) has been tested in clinic and has a very rapid onset of action in patients with diabetes. In order to understand the basis for the rapid insulin absorption, in vitro experiments using analytical ultracentrifugation, protein charge assessment, and light scattering have been performed with this novel human insulin formulation and compared with a commercially available insulin formulation [regular human insulin (RHI)]. METHOD: Analytical ultracentrifugation and dynamic light scattering were used to infer the relative distributions of insulin monomers, dimers, and hexamers in the formulations. Electrical resistance of the insulin solutions characterized the overall net surface charge on the insulin complexes in solution. RESULTS: The results of these experiments demonstrate that the zinc chelating (disodium EDTA) and charge-masking (citrate) excipients used in the formulation changed the properties of RHI in solution, making it dissociate more rapidly into smaller, charge-masked monomer/dimer units, which are twice as rapidly absorbed following subcutaneous injection than RHI (Tmax 60 ± 43 versus 120 ± 70 min). CONCLUSIONS: The combination of rapid dissociation of insulin hexamers upon dilution due to the zinc chelating effects of disodium EDTA followed by the inhibition of insulin monomer/dimer reassociation due to the charge-masking effects of citrate provides the basis for the ultra-rapid absorption of this novel insulin formulation.