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.     

Toxicity of crude oil and dispersed crude oil to ghost shrimp Palaemon serenus and larvae of Australian bass Macquaria novemaculeata

Acute 96‐h LC50 values of the water‐accommodated fraction (WAF) of crude oil, dispersants (Corexit 9500 and Corexit 9527) and dispersed oil combinations were determined in semistatic bioassays with seawater, using the ghost shrimp Palaemon serenus and larval Australian bass (fish) Macquaria novemaculeata. Sodium dodecyl sulphate (SDS) and zinc sulphate were used as reference toxicants and identical bioassays were conducted using these compounds. Total petroleum hydrocarbon (TPH) uptake of shrimp was also measured on the samples taken from the bioassays. The nominal mean (n=4) 96‐h LC50 standard error (SE) values for WAF of crude oil, Corexit 9527, Corexit 9500, dispersed oil (9527) and dispersed oil (9500) were 258,000 ppm (13,000), 49.4 ppm (6.4), 83.1 ppm (5.8), 8.1 ppm (0.3), and 3.6 ppm (0.3) in the shrimp bioassays, respectively. The nominal mean (n=4) 96‐h LC50 (SE) values calculated from the fish larval bioassays were 465,000 ppm (16,000), 14.3 ppm (0.9), 19.8 ppm (1.6), 28.5 ppm (1.4), and 14.1 ppm (2.6) for WAF of crude oil, Corexit 9527, Corexit 9500, dispersed oil (9527), and dispersed oil (9500), respectively. These LC50 values indicate that dispersed oil combinations were significantly more toxic to these organisms than WAF of crude oil. TPH uptake of shrimp increased in correlation to exposure concentrations, and the presence of dispersant made oil more available for shrimp. © 2000 John Wiley & Sons, Inc. Environ Toxicol 15: 91–98, 2000     

Liver antioxidant and plasma immune responses in juvenile golden grey mullet (Liza aurata) exposed to dispersed crude oil

Dispersants are often used after oil spills. To evaluate the environmental cost of this operation in nearshore habitats, the experimental approach conducted in this study exposed juvenile golden grey mullets (Liza aurata) for 48 h to chemically dispersed oil (simulating, in vivo, dispersant application), to dispersant alone in seawater (as an internal control of chemically dispersed oil), to mechanically dispersed oil (simulating, in vivo, natural dispersion), to the water-soluble fraction of oil (simulating, in vivo, an oil slick confinement response technique) and to seawater alone (control condition). Biomarkers such as fluorescence of biliary polycyclic aromatic hydrocarbon (PAH) metabolites, total glutathione liver content, EROD (7-ethoxy-resorufin-O-deethylase) activity, liver antioxidant enzyme activities, liver lipid peroxidation and an innate immune parameter (haemolytic activity of the alternative complement pathway) were measured to assess the toxicity of dispersant application. Significant responses of PAH metabolites and total glutathione content of liver to chemically dispersed oil were found, when compared to water-soluble fraction of oil. As was suggested in other studies, these results highlight that priority must be given to oil slick confinement instead of dispersant application. However, since the same patterns of biomarker responses were observed for both chemically and mechanically dispersed oil, the results also suggest that dispersant application is no more toxic than the natural dispersion occurring in nearshore areas (due to, e.g. waves). The results of this study must, nevertheless, be interpreted cautiously since other components of nearshore habitats must be considered to establish a framework for dispersant use in nearshore areas.     

Toxic effects of chemical dispersant Corexit 9500 on water flea Daphnia magna

In 2010, approximately 2.1 million gallons of chemical dispersants, mainly Corexit 9500, were applied in the Gulf of Mexico to prevent the oil slick from reaching shorelines and to accelerate biodegradation of oil during the Deepwater Horizon oil spill. Recent studies have revealed toxic effects of Corexit 9500 on marine microzooplankton that play important roles in food chains in marine ecosystems. However, there is still little known about the toxic effects of Corexit 9500 on freshwater zooplankton, even though oil spills do occur in freshwater and chemical dispersants may be used in response to these spills. The cladoceran crustacean, water flea Daphnia magna , is a well‐established model species for various toxicological tests, including detection of juvenile hormone‐like activity in test compounds. In this study, we conducted laboratory experiments to investigate the acute and chronic toxicity of Corexit 9500 using D. magna . The acute toxicity test was conducted according to OECD TG202 and the 48 h EC₅₀ was 1.31 ppm (CIs 0.99–1.64 ppm). The reproductive chronic toxicity test was performed following OECD TG211 ANNEX 7 and 21 days LOEC and NOEC values were 4.0 and 2.0 ppm, respectively. These results indicate that Corexit 9500 has toxic effects on daphnids, particularly during the neonatal developmental stage, which is consistent with marine zooplankton results, whereas juvenile hormone‐like activity was not identified. Therefore, our findings of the adverse effects of Corexit 9500 on daphnids suggest that application of this type of chemical dispersant may have catastrophic impacts on freshwater ecosystems by disrupting the key food chain network. Copyright © 2016 John Wiley & Sons, Ltd.     

Chemical dispersant potentiates crude oil impacts on growth, reproduction, and gene expression in Caenorhabditis elegans

The economic, environmental, and human health impacts of the deepwater horizon (DWH) oil spill have been of significant concern in the general public and among scientists. This study employs parallel experiments to test the effects of crude oil from the DWH oil well, chemical dispersant Corexit 9500A, and dispersant-oil mixture on growth and reproduction in the model organism Caenorhabditis elegans. Both the crude oil and the dispersant significantly inhibited the reproduction of C. elegans. Dose-dependent inhibitions of hatched larvae production were observed in worms exposed to both crude oil and dispersant. Importantly, the chemical dispersant Corexit 9500A potentiated crude oil effects; dispersant-oil mixture induced more significant effects than oil or dispersant-alone exposures. While oil-alone exposure and dispersant-alone exposure have none to moderate inhibitory effects on hatched larvae production, respectively, the mixture of dispersant and oil induced much more significant inhibition of offspring production. The production of hatched larvae was almost completely inhibited by several high concentrations of the dispersant-oil mixture. This suggests a sensitive bioassay for future investigation of oil/dispersant impacts on organisms. We also investigated the effects of crude oil/dispersant exposure at the molecular level by measuring the expressions of 31 functional genes. Results showed that the dispersant and the dispersant-oil mixture induced aberrant expressions of 12 protein-coding genes (cat-4, trxr-2, sdhb-1, lev-8, lin-39, unc-115, prdx-3, sod-1, acr-16, ric-3, unc-68, and acr-8). These 12 genes are associated with a variety of biological processes, including egg-laying, oxidative stress, muscle contraction, and neurological functions. In summary, the toxicity potentiating effect of chemical dispersant must be taken into consideration in future crude oil cleanup applications.     

Dispersed crude oil amplifies germ cell apoptosis in Caenorhabditis elegans,followed a CEP-1-dependent pathway

The Deepwater Horizon oil spill is among the most severe environmental disasters in US history. The extent of crude oil released and the subsequent dispersant used for cleanup was unprecedented. The dispersed crude oil represents a unique form of environmental contaminant that warrants investigations of its environmental and human health impacts. Lines of evidence have demonstrated that dispersed oil affects reproduction in various organisms, in a more potent manner than oil- and dispersant-only exposures. However, the action mechanism of dispersed oil remains largely unknown. In this study, we utilized the model organism Caenorhabditis elegans to investigate impacts of dispersed oil exposure on sex cell apoptosis and related gene expressions. Worms were exposed to different diluted levels of crude oil–dispersant (oil–dis) mixtures (20:1, v/v; at 500×, 2,000×, and 5,000× dilutions). The dispersed crude oil significantly increases the number of apoptotic germ cells in treated worms when compared with control at all exposure levels (p < 0.05). Genes involved in the apoptosis pathway were dysregulated, which include ced-13, ced-3, ced-4, ced-9, cep-1, dpl-1, efl-1, efl-2, egl-1, egl-38, lin-35, pax-2, and sir-2.1. Many aberrant expressed genes encoding for core components in apoptosis machinery (cep-1/p53, ced-13/BH3, ced-9/Bcl-2, ced-4/Apaf-1, and ced-3/caspase) displayed consistent expression patterns across all exposure levels. Significantly ced-3/caspase was upregulated at all dispersed oil-treated groups, consistent with the observed apoptosis phenotype. Given cep-1/p53 was activated at all dispersed oil treatments and the germ cell apoptosis was suppressed in the CEP-1 loss of function mutant, the increased apoptosis is likely CEP-1 dependent. In addition, the anti-apoptotic ced-9/Bcl-2 was activated in response to the increase in cell death. This study provides a mechanism understanding of dispersed crude oil-induced reproductive toxicity.     

Impact of dispersant on early life stages of the water flea Daphnia magna and the eastern oyster Crassostrea virginica

In response to the 2010 Deepwater Horizon oil spill, over 1 million gallons of dispersant were applied in Gulf of Mexico offshore waters; Corexit 9500 was the most applied dispersant. The impact on organisms in nearshore and freshwaters has received little scrutiny. Acute 48 h toxicity of Corexit 9500 and a new hyperbranched polyethylenimine (HPEI) dispersant‐like compound were evaluated for the freshwater indicator organism, Daphnia magna and for larval and early spat stages of the Eastern oyster, Crassostrea virginica . For D. magna , Corexit 9500 demonstrated toxicity (EC₅₀ of 0.14 [0.13, 0.15] ppm) similar to the 10‐kDa HPEI (EC₅₀ of 0.16 [0.12, 0.19] ppm). HPEI toxicity increased as a function of molecular weight (1.2 to 750 kDa). The 10 kDa size HPEI was further investigated because it dispersed crude oil with equal effectiveness as Corexit. For Corexit, 100% oyster mortality was detected for the ≤0.2‐mm size classes and mortality >50% for the 0.3‐ and 0.7‐mm size classes at the two greatest concentrations (25 and 50 ppm). HPEI (10 kDa) exhibited low mortality rates (<30%) for all concentrations for all oyster size classes except the 0.1‐mm class. Although mortality rates for this size class were up to 60%, mortality was still less than the mortality caused by Corexit 9500. The low toxicity of HPEI polymers for C. virginica in comparison with Corexit 9500 suggests that HPEI polymers warrant further study.     

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.     

Water-soluble polysaccharides from Ulva intestinalis: Molecular properties,structural elucidation and immunomodulatory activities

Water-soluble sulfated polysaccharides extracted from Ulva intestinalis and fractionated using DEAE Sepharose fast flow column to identify their molecular properties and macrophage cells stimulating activities. Crude and fractions (F₁ and F₂) were formed of neutral sugars (58.7–74.7%), sulfates (6.2–24.5%), uronic acids (4.9–5.9%) and proteins (3.2–10.4%). Different levels of sugar constituents including rhamnose (30.1–39.1%), glucose (39.0–48.4%), galactose (0.0–15.8%), xylose (8.5–11.3) and arabinose (0.0–5.1%). The molecular weight (M_w) of crude and fractionated polysaccharides ranged from 87.1 × 10³ to 194.1 × 10³ (g/mol). Crude polysaccharides were not toxic to RAW264.7 cells and fractions induced cell proliferation. Fraction F₁ stimulated RAW264.7 cells to release considerable amounts of nitric oxide, IL-1β, TNF-α, IL-6, IL-10 and IL-12 cytokines. The main backbone of the most immunostimulating polysaccharide (F₁) was consisted of mixed linkages of (1 → 2)-linked rhamnose and (1 → 2)-linked glucose residues.     

Active avoidance from a crude oil soluble fraction by an Andean paramo copepod

Several oil spills due to ruptures in the pipeline oil systems have occurred at the Andean paramo. A sample of this crude oil was mixed with water from a nearby Andean lagoon and the toxicity of the soluble fraction was assessed through lethal and avoidance assays with a locally occurring copepod (Boeckella occidentalis intermedia). The integration of mortality and avoidance aimed at predicting the immediate decline of copepod populations facing an oil leakage. The 24-h median lethal PAH concentration was 42.7 (26.4–91.6) µg L^?1. In the 12-h avoidance assay, 30 % avoidance was recorded at the highest PAH concentration (19.4 µg L^?1). The mortality at this PAH concentration would be of 25 % and, thus, the population immediate decline would be of 55 %. The inclusion of non-forced exposure testing with the quantification of the avoidance response in environmental risk assessments is, therefore, supported due to underestimation of the lethal assays.     

A test battery approach to the ecotoxicological evaluation of cadmium and copper employing a battery of marine bioassays

Heavy metals are ubiquitous contaminants of the marine environment and can accumulate and persist in sediments. The toxicity of metal contaminants in sediments to organisms is dependent on the bioavailability of the metals in both the water and sediment phases and the sensitivity of the organism to the metal exposure. This study investigated the effects of two metal contaminants of concern (CdCl₂ and CuCl₂) on a battery of marine bioassays employed for sediment assessment. Cadmium, a known carcinogen and widespread marine pollutant, was found to be the least toxic of the two assayed metals in all in vivo tests. However, CdCl₂ was found to be more toxic to the fish cell lines PLHC-1 and RTG-2 than CuCl₂. Tisbe battagliai was the most sensitive species to both metals and the Microtox^® and cell lines were the least sensitive (cadmium was found to be three orders of magnitude less toxic to Vibrio fischeri than to T. battagliai). The sensitivity of Tetraselmis suecica to the two metals varied greatly. Marine microalgae are among the organisms that can tolerate higher levels of cadmium. This hypothesis is demonstrated in this study where it was not possible to derive an EC₅₀ value for CdCl₂ and the marine prasinophyte, T. suecica. Conversely, CuCl₂ was observed to be highly toxic to the marine alga, EC₅₀ of 1.19 mg l^?1. The genotoxic effect of Cu on the marine phytoplankton was evaluated using the Comet assay. Copper concentrations ranging from 0.25 to 2.50 mg l^?1 were used to evaluate the effects. DNA damage was measured as percent number of comets and normal cells. There was no significant DNA damage observed at any concentration of CuCl₂ tested and no correlation with growth inhibition and genetic damage was found.     

Effects of dispersed and adsorbed crude oil on microalgal and bacterial communities of cold seawater

Mesocosm facilities consisting of five 3.5 m³ stainless steel tanks filled with seawater from the St Lawrence Estuary (Québec, Canada) were used to conduct a 2 month experiment under the natural conditions prevailing at the end of the winter in subarctic environments, with seawater temperatures ranging from-1.5°C (surface ice cover) to 3°C. Various oil treatments were simulated in mesocosms: Forties crude oil was chemically dispersed, adsorbed onto an immersed substrate and spilled without any treatment. Total oil concentrations ranged from <1 mgl^-1 (untreated oil) to 44.6 mgl^-1 (dispersed oil). Contrasting with the parent crude oil and dispersed oil, the dissolved phase was enriched with low molecular weight polycyclic aromatic hydrocarbons (PAH). As revealed by pH variations, chlorophyll a contents and degraded pigments, the phytoplankton growth was inhibited early in tanks contaminated with dispersed and adsorbed oil. Although global measurements showed a recovery of the microalgal activity while the dispersed oil was diluted in a flow-through cascade system, the specific composition was quite different from the control, with an increase in small microflagellate species and a marked decline in the diversity of centric diatoms. Small microflagellates also dominated in the heavily oil-contaminated surface microlayer. The growth of viable heterotrophic bacteria (VHB) was immediately stimulated by both dispersed and adsorbed oil (10⁴–10⁵ colony forming units per ml) and oil-degrading bacteria (ODB) reached maximum densities (10² CFU ml^-1) later in the experiment. The adaptation of the indigenous community was assessed using the ODB/VHB ratio, which increased by ten times in the seawater contaminated with dispersed oil. No significant bacterial enhancement was observed in the tank that received untreated oil. No bacterial enrichment was found in the surface microlayer. In sediment traps, the bacterial density increased with the amount of total settling matter and oil residues.     

Essential oil composition, insecticidal and antibacterial activities of Salvia tomentosa Miller

The effectiveness of the essential oil of Salvia tomentosa Miller was assessed on two important pest insects and seven pathogenic bacteria. The essential oil of aerial parts of this plant was hydro-distillated using a clevenger-type apparatus (yield 0.31 % v/w) and constituents were determined using GC–MS analysis. β-pinene (37.28 %) and α-pinene (5.73 %) were the predominant chemical constituents, followed by trans-pinocarveol (3.05 %), myrtenol (2.81 %), caryphyllene oxide (2.68 %), d-camphor (2.08 %). The complete mortality was determined at above doses of 50 μl L^?1 air on Acanthoscelides obtectus (Say) adults. However, the essential oil in concern caused 83.34 and 100 % mortality at higher doses (150 and 200 μl L^?1 air) against Tribolium castaneum (Herbst) adults. The essential oil also revealed significant bacteriostatic and bactericidal activities against Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Micrococcus luteus, Escherichia coli, Enterobacter aerogenes and Yersinia enterocolitica. The essential oil of S. tomentosa in concern can be used as a potential insecticide and bactericidal agents in food applications and agricultural commodities and/or other fields.     

Immunotoxic effects of in vitro exposure of dolphin lymphocytes to Louisiana sweet crude oil and Corexit™

The Deepwater Horizon oil spill was one of the worst environmental disasters on record in the United States. Response efforts to reduce the magnitude of the oil slick included the use of thousands of gallons of the chemical dispersant Corexit™ in surface and deep‐water environments. The immunotoxicity of Louisiana sweet crude oil and the chemical dispersant Corexit was examined using lymphocyte proliferation (LP) and natural killer cell (NK) assays as measures of impact on the adaptive (LP) and innate (NK) immune response in bottlenose dolphins. Study results show that both high‐energy media‐accommodated fractions (MAF) and chemically enhanced MAF (CEMAF) mixtures modulate immune function. Following exposure to Louisiana sweet crude, both B‐ and T‐cell proliferation of white blood cells was increased for all exposure concentrations, compared to control; however, this increase was only significant for the 50% and 100% treatments. In contrast, exposure of white blood cells to the CEMAF mixture significantly decreased both T‐ and B‐cell proliferation in the 25%, 50% and 100% treatments. NK cell activity was enhanced significantly by CEMAF mixtures for the 50% and 100% treatments. The immunosuppression of LP at environmentally relevant concentrations of oil and dispersant suggests that marine mammals may be unable to mount an adequate defense against xenobiotic threats following exposure to oil and dispersant, leaving them more susceptible to disease. In contrast, NK cell activity was significantly enhanced, which may increase an organism's tumor or viral surveillance ability by mounting an enhanced immune response. Copyright © 2016 John Wiley & Sons, Ltd.     

Swimming speed alteration of <Emphasis Type="Italic">Artemia</Emphasis> sp. and <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> as a sub-lethal behavioural end-point for ecotoxicological surveys

In this study, we investigated the possibility to improve a new behavioural bioassay (Swimming Speed Alteration test—SSA test) using larvae of marine cyst-forming organisms: e.g. the brine shrimp Artemia sp. and the rotifer Brachionus plicatilis. Swimming speed was investigated as a behavioural end-point for application in ecotoxicology studies. A first experiment to analyse the linear swimming speed of the two organisms was performed to verify the applicability of the video-camera tracking system, here referred to as Swimming Behavioural Recorder (SBR). A second experiment was performed, exposing organisms to different toxic compounds (zinc pyrithione, Macrotrol^® MT-200, and Eserine). Swimming speed alteration was analyzed together with mortality. The results of the first experiment indicate that SBR is a suitable tool to detect linear swimming speed of the two organisms, since the values have been obtained in accordance with other studies using the same organisms (3.05 mm s^?1 for Artemia sp. and 0.62 mm s^?1 for B. plicatilis). Toxicity test results clearly indicate that swimming speed of Artemia sp. and B. plicatilis is a valid behavioural end-point to detect stress at sub-lethal toxic substance concentrations. Indeed, alterations in swimming speed have been detected at toxic compound concentrations as low as less then 0.1–5% of their LC₅₀ values. In conclusion, the SSA test with B. plicatilis and Artemia sp. can be a good behavioural integrated output for application in marine ecotoxicology and environmental monitoring programs.     

Responses of freshwater mussel (Elliptio complanata) hemocytes exposed in vitro to crude extracts of Microcystis aeruginosa and Lyngbya wollei

Lyngbya wollei is a benthic filamentous cyanobacterium that produces a toxin analogous to the neurotoxic saxitoxin known as lyngbyatoxin (LYNGTX). Microcystis aeruginosa form blooms in the pelagic area of eutrophic lakes and produce a series of potent hepatotoxins–microcystins (MCYST). The aim of this study in vitro study was to examine the difference between the crude extracts of either M. aeruginosa or L. wollei toward the immune system of Elliptio complanata mussels. Freshly isolated hemolymph was plated and exposed to the crude extract of each species at LYNGTX or MCYST equivalent concentrations of 5, 10 and 25 μg/L for 18 h. Immunocompetence was characterized by following changes in hemocyte numbers, metabolic activity (viability), and phagocytosis. Hemocyte counts were not affected, indicating no turnover of hemocytes. Hemocyte metabolic activity was higher in cells exposed to crude extracts of L. wollei. Exposure to L. wollei extracts led to decreased pro-inflammatory precursors such as reactive oxygen species (ROS) and cyclooxygenase (COX) activities. Phagocytosis increased at 25 μg/L for both types of crude extracts. However, hemocytes exposed to crude extracts of M. aeruginosa produced more ROS and COX compared to hemocytes exposed to crude extracts of L. wollei. In conclusion, the data suggest that the crude extract of M. aeruginosa was more toxic than crude extract of L. wollei to mussel hemocytes.     

Evaluation of cyanobacteria toxicity in tropical reservoirs using crude extracts bioassay with cladocerans

During a cyanobacterial bloom in a eutrophic environment, particularly at the end when decomposition occurs, toxic compounds such as the cyanotoxins and the lipopolysaccharides can be released in high concentrations into the water column damaging aquatic organisms. In this work, the effects of this release of toxic compounds during a cyanobacterial bloom were investigated. The acute and chronic toxicity of cyanobacterial crude extracts from two natural blooms in the Barra Bonita and Ibitinga reservoirs (Middle Tietê River, São Paulo State, Brazil) and of a toxic strain cultured in the laboratory were tested. The cladocerans Daphnia similis, Ceriodaphnia dubia and Ceriodaphnia silvestrii were used as test organisms. In the chronic toxicity tests, only a native cladoceran found in Brazilian freshwaters, Ceriodaphnia silvestrii, was used. Microcystins were detected in all cyanobacterial samples. The acute toxicity tests showed that the crude bloom material extract from the Ibitinga Reservoir (48-h EC₅₀ values between 32.6 and 35.8 μg microcystin g^?1 of freeze-dried material) exhibited higher toxicity to cladoceran than did the crude bloom material extract from Barra Bonita Reservoir (48-h EC₅₀ values between 46.0 and 80.2 μg microcystin g^?1 of freeze-dried material). The chronic toxicity test data showed that the three extracts reduced the fecundity of C. silvestrii, and the crude extract of Barra Bonita Reservoir bloom material also affected the survival of this cladoceran. Both acute and chronic tests effectively prognosticated possible changes in the cladoceran population, and probably other components of the biota due to cyanobacterial blooms in natural aquatic ecosystems.     

A computer-controlled whole-body inhalation exposure system for the oil dispersant COREXIT EC9500A

An automated whole-body inhalation exposure system capable of exposing 12 individually housed rats was designed to examine the potential adverse health effects of the oil dispersant COREXIT EC9500A, used extensively during the Deepwater Horizon oil spill. A computer-controlled syringe pump injected the COREXIT EC9500A into an atomizer where droplets and vapor were formed and mixed with diluent air. The aerosolized COREXIT EC9500A was passed into a customized exposure chamber where a calibrated light-scattering instrument estimated the real-time particle mass concentration of the aerosol in the chamber. Software feedback loops controlled the chamber aerosol concentration and pressure throughout each exposure. The particle size distribution of the dispersant aerosol was measured and shown to have a count median aerodynamic diameter of 285 nm with a geometric standard deviation of 1.7. The total chamber concentration (particulate + vapor) was determined using a modification of the acidified methylene blue spectrophotometric assay for anionic surfactants. Tests were conducted to show the effectiveness of closed loop control of chamber concentration and to verify chamber concentration homogeneity. Five automated 5-h animal exposures were performed that produced controlled and consistent COREXIT EC9500A concentrations (27.1 ± 2.9 mg/m(3), mean ± SD).     

Chemical composition and bioactivities of the essential oil from different organs of <Emphasis Type="Italic">Ferula communis</Emphasis> L. growing in Tunisia

The essential oils obtained by the hydrodistillation from the fresh flowers, leaves, stems, and roots of Ferula communis L., growing in Tunisia were analyzed by GC and GC/MS. Thirty-two components were identified in the oil of flowers with camphor (18.3 %), α-pinene (15.3 %), and β-eudesmol (9.3 %) as the main constituents. Twenty-nine compounds were identified in the oil of stems with β-eudesmol (28.1 %), δ-eudesmol (11.1 %), and α-eudesmol (9.6 %) as the main compounds. Twenty compounds were characterized in the oil of roots with dillapiole (7.9 %), guaiol (7.3 %), and spathulenol (6.8 %). In the oil of leaves, α-eudesmol (25.2 %), β-eudesmol (20.7 %), δ-eudesmol (10.1 %), and caryophyllene oxide (7.2 %) were found as the main constituents. This study was undertaken to evaluate the antioxidant activity using DPPH (2,2′-diphenyl-1-picrylhydrazyl), ABTS (2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid), reducing power, and catalase activity. We tested also the antibacterial, cytotoxic, and cholinesterase inhibition properties of the essential oil of different organs of F. communis. The essential oil of the stems showed the highest antioxidant activity (IC₅₀ = 0.03 ± 0.001 mg mL^?1), in DPPH assay and the important result of catalase (303.03 µmol H₂O₂ degraded/min/protein) of F. communis. The antibacterial activity of the oil was determined by micro-well dilution assay. The best results (MIC = 0.156 ± 0.02 mg mL^?1) were exhibited by the essential oil of the leaves of F. Communis against Pseudomonas aeruginosa. Besides, the strongest cytotoxic activity against Hela cells was shown with essential oils’ leaves with an inhibition percentage of 79.05 % at the concentration of 500 µg mL^?1. However, the best inhibition percentage of A 549 cells was detected for essential oils’ leaves with an inhibition percentage of 54.56 % at 250 µg mL^?1. Our finding showed that the essential oil of the flowers was the most active, with 64.623 % of inhibition against butyrylcholinesterase at 10 mg mL^?1 from the incubation time of 30 min.