Biochemical biomarkers in Nile tilapia (Oreochromis niloticus) after short-term exposure to diesel oil, pure biodiesel and biodiesel blends

Fossil fuels such as diesel are being gradually replaced by biodiesel, a renewable energy source, cheaper and less polluting. However, little is known about the toxic effects of this new energy source on aquatic organisms. Thus, we evaluated biochemical biomarkers related to oxidative stress in Nile tilapia (Oreochromis niloticus) after two and seven exposure days to diesel and pure biodiesel (B100) and blends B5 and B20 at concentrations of 0.01 and 0.1 mL L⁻¹. The hepatic ethoxyresorufin-O-deethylase activity was highly induced in all groups, except for those animals exposed to B100. There was an increase in lipid peroxidation in liver and gills in the group exposed to the higher concentration of B5. All treatments caused a significant increase in the levels of 1-hydroxypyrene excreted in the bile after 2 and 7 d, except for those fish exposed to B100. The hepatic glutathione-S-transferase increased after 7 d in animals exposed to the higher concentration of diesel and in the gill of fish exposed to the higher concentration of pure diesel and B5, but decreased for the two tested concentrations of B100. Superoxide dismutase, catalase and glutathione peroxidase also presented significant changes according to the treatments for all groups, including B100. Biodiesel B20 in the conditions tested had fewer adverse effects than diesel and B5 for the Nile tilapia, and can be suggested as a less harmful fuel in substitution to diesel. However, even B100 could activate biochemical responses in fish, at the experimental conditions tested, indicating that this fuel can also represent a risk to the aquatic biota.     

A comparison of acute toxicity of biodiesel, biodiesel blends, and diesel on aquatic organisms

The increased demand of alternative energy sources has created interest in biodiesel and biodiesel blends; biodiesel is promoted as a diesel substitute that is safer, produces less harmful combustion emissions, and biodegrades more easily. Like diesel spills, biodiesel can have deleterious effects on the aquatic environments. The effect of neat biodiesel, biodiesel blends, and diesel on Oncorhynchus mykiss and Daphnia magna was evaluated using acute toxicity testing. Static nonrenewal bioassays of freshwater organisms containing B100, B50, B20, B5, and conventional diesel fuel were used to compare the acute effects of biodiesel to diesel. Mortality was the significant end point measured in this study; percent mortality and lethal concentration (LC50) at different exposure times were determined from the acute toxicity tests performed. Trials were considered valid if the controls exhibited > 90% survival. Based on percentage of mortality and LC50 values, a toxicity ranking of fuels was developed.     

Influence of constant and fluctuating temperature on biodegradation rates of fish biodiesel blends contaminating Alaskan sand

Bioremediation of sandy soil contaminated with fish-biodiesel, conventional diesel, and blends of both was studied in microcosm experiments at different temperatures, simulating the subarctic environment. While distinct lag, exponential, and stationary phases were observed at 20 °C, degradation at 6 °C was slow and the lag phase continued throughout the 4-week experiment. A three-phase 1st order kinetic model successfully described respiration at 20 °C, a one-phase model was sufficient at 6 °C. For temperatures fluctuating between ∼6 and ∼20 °C, higher than expected microbial activity persisted at 6 °C for several days, due to the presence of active cultures, even though the soil temperature closely followed the air temperature. At 20 °C, respiration peaked already after 1 week, and 18-51% of the initially added fuel was mineralized within 4 weeks, whereby degradation was higher at higher biodiesel percentages. Biodiesel addition accelerated mineralization of blends with regular diesel beyond expectations. In blends with 20% biodiesel, the degradation rate constant was twice as high as for conventional diesel. These synergistic effects are likely due to an active microbial population. Addition of biodiesel to conventional diesel could reduce the impact of diesel spills.     

Toxic effects of Hydrilla verticillata exposed to toluene, ethylbenzene and xylene and safety assessment for protecting aquatic macrophytes

Little information is available about the toxicity of toluene, ethylbenzene and xylene acting on macrophytes, and their toxicity data are rarely used in regulation and criteria decisions. The results extended the knowledge on toxic effects of toluene, ethylbenzene and xylene on aquatic plants. The responses of Hydrilla verticillata to these pollutants were investigated. Chlorophyll levels, lipid peroxidation, and antioxidant enzymes (superoxide dismutase and guaiacol peroxidase) showed diverse responses at different concentrations of toluene, ethylbenzene and xylene. The linear regression analyses were performed respectively, suggesting the concentrations of toluene, ethylbenzene and xylene expected to protect aquatic macrophytes were 7.30 mg L⁻¹, 1.15 mg L⁻¹ and 2.36 mg L⁻¹, respectively. This study emphasized that aquatic plants are also sensitive to organic pollutants as fishes and zooplanktons, indicating that macrophytes could be helpful in predicting the toxicity of these pollutants and should be considered in regulation and criteria decisions for aquatic environment protection.     

Atrazine promotes biochemical changes and DNA damage in a Neotropical fish species

The effects of Atrazine, an herbicide used worldwide and considered as a potential contaminant in aquatic environments, were assessed on the Neotropical fish Prochilodus lineatus acutely (24 and 48 h) exposed to 2 or 10 μg L⁻¹ of atrazine by using a set of biochemical and genetic biomarkers. The following parameters were measured in the liver: activity of the biotransformation enzymes ethoxyresorufin-O-deethylase (EROD) and glutathione S transferase (GST), antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), content of reduced glutathione (GSH), generation of reactive oxygen species (ROS) and occurrence of lipid peroxidation (LPO); in brain and muscle the activity of acetylcholinesterase (AChE) and DNA damage (comet assay) on erythrocytes, gills and liver cells. A general decreasing trend on the biotransformation and antioxidant enzymes was observed in the liver of P. lineatus exposed to atrazine; except for GR, all the other antioxidant enzymes (SOD, CAT and GPx) and biotransformation enzymes (EROD and GST) showed inhibited activity. Changes in muscle or brain AChE were not detected. DNA damage was observed in the different cell types of fish exposed to the herbicide, and it was probably not from oxidative origin, since no increase in ROS generation and LPO was detected in the liver. These results show that atrazine behaves as enzyme inhibitor, impairing hepatic metabolism, and produces genotoxic damage to different cell types of P. lineatus.     

Effects of biodiesel on emissions of regulated air pollutants and polycyclic aromatic hydrocarbons under engine durability testing

An 80,000-km durability test was performed on two engines using diesel and biodiesel (methyl ester of waste cooking oil) as fuel in order to examine emissions resulting from the use of biodiesel. The test biodiesel (B20) was blended with 80% diesel and 20% methyl ester derived from waste cooking oil. Emissions of regulated air pollutants, including CO, HC, NO_x, particulate matter (PM) and polycyclic aromatic hydrocarbons (PAHs) were measured at 20,000-km intervals. The identical-model engines were installed on a standard dynamometer equipped with a dilution tunnel used to measure the pollutants. To simulate real-world driving conditions, emission measurements were made in accordance with the United States Environmental Protection Agency (USEPA) FTP transient cycle guidelines. At 0 km of the durability test, HC, CO and PM emission levels were lower for the B20 engine than those for diesel. After running for 20,000 km and longer, they were higher. However, the deterioration coefficients for these regulated air pollutants were not statistically higher than 1.0, implying that the emission factors do not increase significantly after 80,000 km of driving. Total (gaseous+particulate phase) PAH emission levels for both B20 and diesel decreased as the driving mileage accumulated. However, for the engine using B20 fuel, particulate PAH emissions increased as engine mileage increased. The average total PAH emission factors were 1097 and 1437 μg bhp h⁻¹ for B20 and diesel, respectively. For B20, the benzo[a]pyrene equivalence emission factors were 0.77, 0.24, 0.20, 7.48, 5.43 and 14.1 μg bhp h⁻¹ for 2-, 3-, 4-, 5-, 6-ringed and total PAHs. Results show that B20 use can reduce both PAH emission and its corresponding carcinogenic potency.     

Regulated emissions from biodiesel fuels from on/off-road applications

This research is one of the largest studies of biodiesel in both on-road and off-road uses. The testing was conducted for the military and encompassed a wide range of application types including two medium-duty trucks, two Humvees, a heavy heavy-duty diesel truck, a bus, two stationary backup generators (BUGs), a forklift, and an airport tow vehicle. The full range of fuels tested included a California ultra-low sulfur diesel (ULSD) fuel, different blend ratios of two different yellow-grease biodiesels and one soy-based biodiesel, JP-8, and yellow-grease biodiesel blends with two different NO_x reduction additives. The B20-YGA, B20-YGB, and B20-Soy did not show trends relative to ULSD that were consistent over all applications tested. Higher biodiesel blends were tested on only one vehicle, but showed a tendency for higher total hydrocarbons (THC) and carbon monoxide (CO) emissions and lower particulate matter (PM) emissions. The JP-8 showed increases in THC and CO relative to the ULSD.     

Regulated and unregulated emissions from a diesel engine fueled with biodiesel and biodiesel blended with methanol

Experiments were carried out on a diesel engine operating on Euro V diesel fuel, pure biodiesel and biodiesel blended with methanol. The blended fuels contain 5%, 10% and 15% by volume of methanol. Experiments were conducted under five engine loads at a steady speed of 1800 rev min⁻¹ to assess the performance and the emissions of the engine associated with the application of the different fuels. The results indicate an increase of brake specific fuel consumption and brake thermal efficiency when the diesel engine was operated with biodiesel and the blended fuels, compared with the diesel fuel. The blended fuels could lead to higher CO and HC emissions than biodiesel, higher CO emission but lower HC emission than the diesel fuel. There are simultaneous reductions of NO_x and PM to a level below those of the diesel fuel. Regarding the unregulated emissions, compared with the diesel fuel, the blended fuels generate higher formaldehyde, acetaldehyde and unburned methanol emissions, lower 1,3-butadiene and benzene emissions, while the toluene and xylene emissions not significantly different.     

Characteristics of trans,trans-2,4-decadienal and polycyclic aromatic hydrocarbons in exhaust of diesel engine fueled with biodiesel

The use of biodiesel fuel as a substitute for fossil fuel in diesel engines has received increasing attention in recent years. This study is the first to investigate and compare the characteristics of mutagenic species, trans,trans-2,4-decadienal (tt-DDE), and polycyclic aromatic hydrocarbons (PAHs) in the diluted exhaust of diesel engines operated with diesel and biodiesel blend fuels. An engine of current design was operated on a dynamometer consistent with the US federal test procedure transient-cycle specifications. Petroleum diesel and a blend of petroleum diesel and biodiesel (B20) were tested. Exhaust sampling was carried out on diluted exhaust in a dilution tunnel with a constant-volume sampling system. Concentrations of tt-DDE and PAHs were analyzed by GC/MS. Although average PAH emission factors decreased from 1403 to 1051 μg bhp-h⁻¹, the results show that tt-DDE is evidently generated (1.28 μg bhp-h⁻¹) in the exhaust of diesel engine using B20 as fuel. This finding suggests that tt-DDE emission from the use of biodiesel should be taken into account in characterization and health-risk assessment. The results also show that tt-DDE is depleted in the diesel engine combustion process and the existence of tt-DDE in biodiesel is the major source of tt-DDE emission. The distribution of tt-DDE in the particulate phase is 55.3% under this study's sampling conditions. For diesel and B20, PAH phase distributions have similar trends. Lower molecular weight PAHs predominate in gaseous phase for both diesel and B20. Cold-start driving has higher tt-DDE and PAH emission factors, as well as a higher percentage of tt-DDE in particulate phase, than for warm-start driving.     

Ecotoxicological responses of the earthworm Eisenia fetida exposed to soil contaminated with HHCB

Although polycyclic musks have been shown to cause lethal and sub-lethal effects on organisms, their biochemical toxicity to earthworms is not well understood. In the current study, we investigated the responses of antioxidant systems and lipid peroxidation after exposing Eisenia fetida to soil contaminated with 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethyl-cyclopenta-γ-2-benzopyran (HHCB). Significant increase in lipid peroxidation level was observed on day 14 at two high concentrations, 50 and 100 mg kg⁻¹. Among antioxidant enzymes, the primary response to chronic HHCB exposure can be attributed to superoxide dismutase (SOD) and catalase (CAT). Of the two enzymes, SOD exhibited more sensitive response to HHCB stress. In addition, these two enzymes could have a combined effect on fighting damage by reactive oxygen species, evidenced by a marked relationship between lipid peroxidation and enzyme activity. On the other hand, dose-dependent inhibition of peroxidase (POD) activity has been observed throughout the test. The results suggest that the variations in investigated parameters of E. fetida could be used as responsive biomarkers for oxidative stress caused by HHCB in a soil environment.     

Effects of different inorganic arsenic species in Cyprinus carpio (Cyprinidae) tissues after short-time exposure: Bioaccumulation, biotransformation and biological responses

Differences in the toxicological and metabolic pathway of inorganic arsenic compounds are largely unknown for aquatic species. In the present study the effects of short-time and acute exposure to As^III and As^V were investigated in gills and liver of the common carp, Cyprinus carpio (Cyprinidae), measuring accumulation and chemical speciation of arsenic, and the activity of glutathione-S-transferase omega (GST Ω), the rate limiting enzyme in biotransformation of inorganic arsenic. Oxidative biomarkers included antioxidant defenses (total glutathione-S-transferases, glutathione reductase, glutathione, and glucose-6-phosphate dehydrogenase), total scavenging capacity toward peroxyl radicals, reactive oxygen species (ROS) measurement and lipid peroxidation products. A marked accumulation of arsenic was observed only in gills of carps exposed to 1000 ppb As^V. Also in gills, antioxidant responses were mostly modulated through a significant induction of glucose-6-phosphate dehydrogenase activity which probably contributed to reduce ROS formation; however this increase was not sufficient to prevent lipid peroxidation. No changes in metal content were measured in liver of exposed carps, characterized by lower activity of GST Ω compared to gills. On the other hand, glutathione metabolism was more sensitive in liver tissue, where a significant inhibition of glutathione reductase was concomitant with increased levels of glutathione and higher total antioxidant capacity toward peroxyl radicals, thus preventing lipid peroxidation and ROS production. The overall results of this study indicated that exposure of C. carpio to As^III and As^V can induce different responses in gills and liver of this aquatic organism.     

Histopathological and biochemical alternations of the heart induced by acute cadmium exposure in the freshwater crab Sinopotamon yangtsekiense

Cadmium (Cd) is a highly toxic element in water. Its toxicity has been attributed to oxidative stress mediated by free radicals. Here we investigated the effects of Cd on the histopathology, antioxidant enzymes and lipid peroxidation of crustacean heart. The freshwater crabs Sinopotamon yangtsekiense were exposed to different concentrations of Cd for 1, 3, 5 and 7 d. After exposure, histological abnormalities were discovered, including myocardial edema, vacuolar and vitreous degeneration, and infiltration of inflammatory cells. Additionally, alterations in nuclei, mitochondria, rough endoplasmic reticulum as well as myofibrils were observed. Meanwhile, superoxide dismutase (SOD) activity was significantly increased after Cd exposure. Catalase (CAT) activity was only increased in the group exposed to 14.50 mg L⁻¹ Cd on day 5 and decreased with increasing Cd concentration and exposure time. Glutathione peroxidase (GPx) activity was increased in groups treated with 29.00, 58.00 and 116.00 mg L⁻¹ on days 1 and 3, and decreased thereafter. Besides, malondialdehyde (MDA) levels were significantly increased after 3 d of Cd exposure at all the indicated concentrations. These results showed that acute Cd exposure led to harmful effects on the histology of crab heart, which are most likely linked to Cd-induced oxidative stress.     

Hydrocarbon emissions speciation in diesel and biodiesel exhausts

Diesel engine emissions are composed of a long list of organic compounds, ranging from C₂ to C₁₂₊, and coming from the hydrocarbons partially oxidized in combustion or produced by pyrolisis. Many of these are considered as ozone precursors in the atmosphere, since they can interact with nitrogen oxides to produce ozone under atmospheric conditions in the presence of sunlight. In addition to problematic ozone production, Brookes, P., and Duncan, M. [1971. Carcinogenic hydrocarbons and human cells in culture. Nature.] and Heywood, J. [1988. Internal Combustion Engine Fundamentals.Mc Graw-Hill, ISBN 0-07-1000499-8.] determined that the polycyclic aromatic hydrocarbons present in exhaust gases are dangerous to human health, being highly carcinogenic.The aim of this study was to identify by means of gas chromatography the amount of each hydrocarbon species present in the exhaust gases of diesel engines operating with different biodiesel blends. The levels of reactive and non-reactive hydrocarbons present in diesel engine exhaust gases powered by different biodiesel fuel blends were also analyzed.Detailed speciation revealed a drastic change in the nature and quantity of semi-volatile compounds when biodiesel fuels are employed, the most affected being the aromatic compounds. Both aromatic and oxygenated aromatic compounds were found in biodiesel exhaust. Finally, the conservation of species for off-side analysis and the possible influence of engine operating conditions on the chemical characterization of the semi-volatile compound phase are discussed.The use of oxygenated fuel blends shows a reduction in the Engine-Out emissions of total hydrocarbons. But the potential of the hydrocarbon emissions is more dependent on the compositions of these hydrocarbons in the Engine-Out, to the quantity; a large percent of hydrocarbons existing in the exhaust, when biodiesel blends are used, are partially burned hydrocarbons, and are interesting as they have the maximum reactivity, but with the use of pure biodiesel and diesel, the most hydrocarbons are from unburned fuel and they have a less reactivity. The best composition in the fuel, for the control of the hydrocarbon emissions reactivity, needs to be a fuel with high-saturated fatty acid content.     

Biomarker responses as indication of contaminant effects in Oreochromis niloticus

The current study investigated oxidative stress parameters (enzymes activities, metallothionein content and lipid peroxidation) in freshwater fish, Oreochromis niloticus, tilapia exposure to Monjolinho River (in 4 months of year: January, April, July and November). One critical site in Monjolinho River (site B) was assessed in comparison to a reference site (site A). Water pH and oxygen concentration was lower than that recommended by CONAMA (Brazilian National Environmental Committee), resolution 357/2005 for protection of aquatic communities, and ammonium and the metals Cu, Zn, Mn and Fe (on all months) concentrations were higher than the maximum concentration recommended. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were significantly decreased in liver and muscle in tilapia from Monjolinho River, throughout the year, in relation to reference except in gills that SOD activity increased. Glutathione S-transferase (GST) activity was significantly increased in liver of the tilapia from Monjolinho River in all sites, in relation to reference except in gills that GST activity increased in July and decreased in November, suggesting that GST activity could be induced to neutralize the pollutants toxicity. On the other hand, GST activity was significantly decreased in white muscle indicating a toxic effect of pollutants, resulting in a decreased ability of tilapia to perform defense reactions associated to GSTs. The decrease of catalase (CAT) activity in gills of the O. niloticus together with the increase of SOD activity, could explain the increased lipid peroxidation (LPO) level in this organ. Metallothionein levels in liver and gills were significantly high in all sites. Results indicate that the exposure to metals caused severe damage to tissues; despite the consensually assumed antioxidant induction as a sign of exposure to contaminants the effects seem in part to be mediated by suppression of antioxidant system with SOD, CAT and GPx as potential candidates for tissues toxicity biomarkers of pollutants.     

Effects of diesel/biodiesel blends on regulated and unregulated pollutants from a passenger vehicle operated over the European and the Athens driving cycles

This paper presents the regulated and unregulated exhaust emissions of a diesel passenger vehicle, operated with low sulphur automotive diesel and soy methyl ester blends. Emission and fuel consumption measurements were conducted under real driving conditions (Athens Driving Cycle, ADC) and compared with those of a modified New European Driving Cycle (NEDC) using a chassis dynamometer. A Euro II compliant diesel vehicle was used in this study, equipped with an indirect injection diesel engine, fuelled with diesel fuel and biodiesel blends at proportions of 5, 10, and 20% respectively. Unregulated emissions of 11 polycyclic aromatic hydrocarbons (PAHs), 5 nitro-PAHs, 13 carbonyl compounds (CBCs) and the soluble organic fraction (SOF) of the particulate matter were measured. Qualitative hydrocarbon analysis was also performed on the SOF. Regulated emissions of NO_x, CO, HC, CO₂, and PM were also measured over the two test cycles. It was established that some of the emissions measured over the (hot-start) NEDC differed from the real-world cycle. Significant differences were also observed in the vehicle's fuel consumption between the two test cycles. The addition of biodiesel reduced the regulated emissions of CO, HC and PM, while an increase in NO_x was observed over the ADC. Carbonyl emissions, PAHs and nitro-PAHs were reduced with the addition of biodiesel over both driving cycles.     

Histopathological effects of copper and lithium in the ramshorn snail, Marisa cornuarietis (Gastropoda, Prosobranchia)

The aim of this study was to determine and quantify effects of copper and lithium in tissues of early juveniles of the ramshorn snail, Marisa cornuarietis. For this purpose, hatchlings of M. cornuarietis were exposed for 7 days to a range of five different sublethal concentrations of copper (5, 10, 25, 50, and 75 μg Cu²⁺ L⁻¹) and lithium (50, 100, 200, 1000, and 5000 μg Li⁺ L⁻¹). Both metals changed the tissue structure of epidermis, hepatopancreas, and gills, varying between slight and strong reactions, depending on the copper and lithium concentration. The histopathological changes included alterations in epithelial and mucous cells of the epidermis, swelling of hepatopancreatic digestive cells, alterations in the number of basophilic cells, abnormal apices of digestive cells, irregularly shaped cilia and changes in the amount of mucus in the gills. The most sensible organ in M. cornuarietis indicating Cu or Li pollution is the hepatopancreas (LOECs were 10 μg Cu²⁺ L⁻¹, or 200 μg Li⁺ L⁻¹). In epidermis, mantle and gills relevant effects occurred with higher LOECs (50 μg Cu²⁺ L⁻¹, or 1000 μg Li⁺ L⁻¹). Base on LOECs, our results indicated that histopathological endpoints are high sensitivity to copper and lithium compared to endpoints for embryonic developmental toxicity.     

Effect of SME biodiesel blends on PM2.5 emission from a heavy-duty engine

To explore the effect of biodiesel and sulfur content on PM_2.5 emissions, engine dynamometer tests were performed on a Euro II engine to compare the PM_2.5 emissions from four fuels: two petroleum diesel fuels with sulfur contents of 50 and 100 ppm respectively, and two B20 fuels in which soy methyl ester (SME) biodiesel was added to each of the above mentioned petroleum diesel fuels (v/v: 80%/20% for petroleum diesel and SME respectively). Gaseous pollutants and PM_2.5 emissions were sampled with an AVL AMA4000 and Model 130 High-Flow Impactor (MSP Corp). Measurements were made of the PM_2.5 mass, organic carbon (OC), elemental carbon (EC) and the water-soluble ion distribution. The results showed that PM_2.5 emissions decreased with lower sulfur content or blending with SME biodiesel, and the decrease would be more by applying both two methods together. Particles of approximately 0.13 μm contributed 48–83% of PM_2.5 emissions. The impact of sulfur content on this percentage was different for low and high engine speed. The majority of PM_2.5 was comprised of OC and EC, and the carbon emission rate had the same trend as PM_2.5. Since the EC abatement of B20 was larger than OC, the OC/EC ratio of B20 was always larger than that of petroleum diesel. For petroleum diesel, the OC/EC increased with sulfur content, which was not the case for B20. The SO₄^2? had highest emission rate in the water-soluble ions of PM.     

Determination of hypoxia and dietary copper mediated sub-lethal toxicity in carp, Cyprinus carpio, at different levels of biological organisation

Hypoxic events frequently occur in the aquatic environment in association with micro pollutants, including heavy metals. Only a few studies are however available on the uptake and biological responses of heavy metals under hypoxic conditions. To elucidate the phenomenon, mirror carp Cyprinus carpio L. (16.13-16.22 g) were exposed chronically to dietary copper (Cu; 250 and 500 mg kg dry wt.⁻¹) for 30 d under normoxic (8.25 mg O₂ L⁻¹) and hypoxic (∼3 mg O₂ L⁻¹) conditions and adopting an integrated approach, sub-lethal biomarker responses were determined at different levels of biological organisation. Level of oxidative DNA damage (as determined by modified Comet assay) showed strong significant difference following exposure to dietary Cu level under normoxic (1.6-fold) as well as under hypoxic condition at both Cu levels (2.1 and 2.5-folds respectively). Significant difference was also observed for haematological parameters (i.e. increased red and white blood cells, haematocrit value and haemoglobin concentration). Quantitative histology revealed alterations in tissues (i.e. liver and gills) for hypoxic and all dietary Cu treatment groups under both normoxic and hypoxic conditions suggesting a compensatory response to these organs (p < 0.05). The order of Cu accumulation in tissues (as determined by ICP-OES) was liver > intestine > kidney > gill. Interestingly, SGR under both normoxic and hypoxic conditions reduced with elevating Cu levels (p = 0.019). Overall, the results provide evidence for enhanced toxicological responses in fish following exposure to Cu either alone or in combination with hypoxic condition and lends support to the evolving viewpoint that many water quality guidelines should be revisited in terms of new ecotoxicological criteria.     

Levels of organochlorine pesticides and polychlorinated biphenyls in the critically endangered Iberian lynx and other sympatric carnivores in Spain

Accumulation of organochlorine compounds is well studied in aquatic food chains whereas little information is available from terrestrial food chains. This study presents data of organochlorine levels in tissue and plasma samples of 15 critically endangered Iberian lynx (Lynx pardinus) and other 55 wild carnivores belonging to five species from three natural areas of Spain (Doñana National Park, Sierra Morena and Lozoya River) and explores their relationship with species diet. The Iberian lynx, with a diet based on the consumption of rabbit, had lower PCB levels (geometric means, plasma: <0.01 ng mL⁻¹, liver: 0.4 ng g⁻¹ wet weight, fat: 87 ng g⁻¹ lipid weight) than other carnivores with more anthropic and opportunistic foraging behavior, such as the red fox (Vulpes vulpes; plasma: 1.11 ng mL⁻¹, liver: 459 ng g⁻¹, fat: 1984 ng g⁻¹), or with diets including reptiles at higher proportion, such as the Egyptian mongoose (Herpestes ichneumon; plasma: 7.15 ng mL⁻¹, liver: 216 ng g⁻¹, fat: 540 ng g⁻¹), or the common genet (Genetta genetta; liver: 466 ng g⁻¹, fat: 3854 ng g⁻¹). Chlorinated pesticides showed interspecific variations similar to PCBs. Organochlorine levels have declined since the 80s in carnivores from Doñana National Park, but PCB levels are still of concern in Eurasian otters (Lutra lutra; liver: 3873-5426 ng g⁻¹) from the industrialized region of Madrid.     

Comparative acute freshwater hazard assessment and preliminary PNEC development for eight fluorinated acids

Short-term 48, 72 and 96-h aquatic toxicity tests were conducted to evaluate the acute toxicity of eight fluorinated acids to the cladoceran, Daphnia magna, the green alga, Pseudokirchneriella subcapitata, and the rainbow trout, Oncorhynchus mykiss or the fathead minnow, Pimephales promelas. The eight fluorinated acids studied were tridecafluorohexyl ethanoic acid (6:2 FTCA), heptadecafluorooctyl ethanoic acid (8:2 FTCA), 2H-dodecafluoro-2-octenoic acid (6:2 FTUCA), 2H-hexadecafluoro-2-decenoic acid (8:2 FTUCA), 2H,2H,3H,3H-undecafluoro octanoic acid (5:3 acid), 2H,2H,3H,3H-pentadecafluoro decanoic acid (7:3 acid), n-perfluoropentanoic acid (PFPeA) and n-perfluorodecanoic acid (PFDA). The results of the acute toxicity tests conducted during this study suggest that the polyfluorinated acids, 8:2 FTCA, 8:2 FTUCA, 6:2 FTCA, 6:2 FTUCA, 7:3 acid and 5:3 acid, and the perfluorinated acids PFPeA and PFDA, are generally of low to medium concern based on evaluation of their acute freshwater toxicity (EC/LC50s typically between 1 and >100 mg L⁻¹) using the USEPA TSCA aquatic toxicity evaluation paradigm. For the polyfluorinated acids, aquatic toxicity generally decreased as the number of fluorinated carbons decreased and as the overall carbon chain length decreased from 12 to 8. Acute aquatic toxicity of the 5 and 10 carbon perfluorocarboxylic acids (EC/LC50s between 10.6 and >100 mg L⁻¹) was greater or similar to that of the 6-9 carbon perfluorocarboxylic acids (EC/LC50s > 96.5 mg L⁻¹). This study also provides the first report of the acute aquatic toxicity of the 5:3 acid (EC/LC50s of 22.5 to >103 mg L⁻¹) which demonstrated less aquatic toxicity than the 7:3 acid (EC/LC50s of 0.4-32 mg L⁻¹). The cladoceran, D. magna and the green alga, P. subcapitata had generally similar EC50 values for a given substance while fish were typically equally or less sensitive with the exception that PFPeA was most toxic to fish. Predicted no-effect concentrations (PNECs) were estimated using approaches consistent with REACH guidance and when compared with available environmental concentrations, these PNECs suggest that the fluorinated acids tested pose little risk for aquatic organisms.