Development of a laboratory exposure system using marine fish to carry out realistic effect studies with produced water discharged from offshore oil production

A biotest system for environmentally realistic exposure of fish to produced water (PW) was developed and tested. Authentic PW was collected at an oil production platform in the North Sea and preserved by freezing in multiple aliquots a 25 L. After transport to the test laboratory onshore, daily PW aliquots were thawed, homogenised and administered to the test fish, Atlantic cod (Gadus morhua), in two diluted exposure concentrations, 0.1% and 0.5%, during a 15 d period, using a continuous flow-through exposure setup. Positive control groups were exposed to two crude oil treatments for comparison. Chemical analyses showed that alkylphenol (AP) and PAH concentrations in PW exposure waters were very low. Observations of significantly increased AP and PAH metabolite levels in PW exposed fish demonstrated the suitability of the biotest system for its use in biological exposure/effect studies of PW, and it also demonstrated the sensitivity of bile metabolites as PW exposure markers in fish. The relevance of the biotest system for PW effect studies and for validating modelled environmental risk estimates of PW dischargers from offshore oil production is discussed.     

The Arctic is no longer put on ice: Evaluation of Polar cod (Boreogadussaida) as a monitoring species of oil pollution in cold waters

The withdrawing Arctic ice edge will facilitate future sea transport and exploration activities in the area, which calls for the establishment of relevant cold water monitoring species. The present study presents first results of field baseline levels for core oil pollution biomarkers in Polar cod (Boreogadussaida) sampled from pristine, Arctic waters. Furthermore, biomarker response levels were characterized in controlled laboratory exposure experiments running over 2 weeks. Fish exposed to a simulated petrogenic spill (1 ppm dispersed, crude oil) exhibited elevated hepatic EROD activity, bile PAH-metabolites, and hepatic DNA-adducts, whereas male individuals exposed to simulated produced water (30 ppb nonylphenol) exhibited a strong induction of plasma vitellogenin. In conclusion, the results demonstrated low and robust biomarker baseline levels that were clearly different from exposure responses. In combination with its high abundance and circumpolar distribution, the Polar cod seems well qualified for oil pollution monitoring in Arctic waters.     

In situ biomonitoring of caged, juvenile Chinook salmon (Oncorhynchus tshawytscha) in the Lower Duwamish Waterway

Contaminated sediments may have wide-ranging impacts on human and ecological health. A series of in situ caged exposure studies using juvenile Chinook salmon was conducted in the Lower Duwamish Waterway (LDW). Chemical analysis of sediment, water, and fish tissue were completed. Additionally, in 2004, DNA adducts in hepatic and gill tissues were measured. Gills contained significantly higher DNA adducts at stations B2 and B4, prompting further analysis of gills in 2006 and 2007. Fluorescent aromatic compounds (FACs) in bile, and CYP1A1 in hepatic tissue were also measured during 2006 and 2007, respectively. FACs in field-caged fish were comparable or significantly higher than wild-caught fish LDW fish and significantly higher than lab fish after only 8-10 days, demonstrating the equivalency of exposure to that of migrating salmon. Furthermore, selected biomarkers appear to be capable of detecting spikes in contamination between sampling years, emphasizing the need for multiple year data collection.     

Bioconcentration of selected endocrine disrupting compounds in the Mediterranean mussel, Mytilus galloprovincialis

The occurrence of three endocrine disrupting compounds, 4-n-nonylphenol, triclosan and bisphenol A, was investigated in different bivalves originating from the Aegean Sea (Greece). The bioconcentration potential of these compounds was studied using the Mediterranean mussel (Mytilus galloprovincialis). Tissue samples were extracted by sonication. Analysis was performed by gas chromatography-mass spectrometry. According to the field survey results, the average concentrations of 4-n-nonylphenol, triclosan and bisphenol A, were 158, 461 and 404 ng g⁻¹ (dry weight), respectively. During 28 days of exposure at 300 ng L⁻¹, the tissue concentrations of compounds were constantly increased. Steady state was not observed up to the end of the experiment. Kinetic bioconcentration factors varied from 1.7 (4-n-nonylphenol and triclosan) to 4.5 L g⁻¹ (bisphenol A). Following exposure, mussels were relocated to clean water for 28 days. This experiment revealed that depuration rates for all of the target compounds were lower than uptake rates. The biological half-lives of each compound ranged between 12 days (triclosan) and 26 days (bisphenol A).     

Bioconcentration of the anionic surfactant linear alkylbenzene sulfonate (LAS) in the marine shrimp Palaemonetes varians: A radiotracer study

Uptake and depuration kinetics of dissolved [¹⁴C]C₁₂-6-linear alkylbenzene sulfonate (LAS) were determined in the shrimp Palaemonetes varians using environmentally relevant exposure concentration. The shrimp concentrated LAS from seawater with a mean BCF value of 120 L kg⁻¹ after a 7-day exposure. Uptake biokinetics were best described by a saturation model, with an estimated BCF_ss, of 159 ± 34 L kg⁻¹, reached after 11.5 days. Shrimp weight influenced significantly BCF value with smaller individuals presenting higher affinity to LAS. To the light of a whole body autoradiography, major accumulation of LAS occurred in the cephalothorax circulatory system (gills, heart, hepatopancreas) and ocular peduncle, but not in the flesh, limiting potential transfer to human consumers. LAS depuration rate constant value of the shrimp was 1.18 ± 0.08 d⁻¹ leading to less than 1% of remaining LAS in its tissues after 8 days of depuration.     

Testes specific accumulation of tributyltin in turbot Scophthalmus maximus from the southern Baltic Sea

Concentrations of tributyltin (TBT) and its breakdown products, dibutyltin (DBT) and monobutyltin (MTB), were measured in tissues of male turbot Scophthalmus maximus from the Gulf of Gdańsk (GDA) and the Pomeranian Bay (POM) in the southern Baltic Sea. The fish (n = 206) were collected during the spawning seasons of 2008-2009. All specimens were at the same maturity stage and in full breeding condition. The ripe testes of the turbot appeared to be a target tissue for TBT accumulation. We report that TBT is extensively eliminated at the specific whole-body level and that the males from TBT-contaminated location (GDA) reach this specific level at 23-24 cm in total length.     

Non-food-chain transfer of sediment-associated persistent organic pollutants to a marine benthic fish

We investigated the transfer of persistent organic pollutants (POPs) from unspiked bottom sediment to a benthic marine fish, marbled sole (Pleuronectes yokohamae), via non-food-chain pathways, i.e., via sediment particles and water column. One-year-old sole were held for 28 days in an exposure tank with bottom sediment or in a control tank. o,p′-DDE and tri- to penta-chlorobiphenyls were transferred from the sediment to the fish via non-food-chain pathways, as demonstrated by concentrations in the exposed fish at 2.5-30 times the control levels. A model analysis based on first-order kinetic equations indicated that the overall rate constant of transfer of these compounds from sediment to fish was generally lower than that from food (median of ratio, 0.48). It also suggested that relatively high concentrations of the other POPs in the food and the longer times necessary for them to reach a steady state masked any transfer of them from the sediment.     

Biomarker responses in Atlantic cod (Gadus morhua) exposed to produced water from a North Sea oil field: laboratory and field assessments

Biological markers of produced water (PW) exposure were studied in Atlantic cod (Gadus morhua) in both laboratory and field experiments, using authentic PW from a North Sea oil field. In the laboratory study, the PW exposure yielded significantly elevated levels of metabolites of polycyclic aromatic hydrocarbons (PAHs) and alkylphenols (APs) in bile even at the lowest exposure dose (0.125% PW). Other biomarkers (hepatic CYP1A induction and DNA adduct formation) responded at 0.25% and 0.5% PW concentrations. In the field study, bile metabolite markers and hepatic CYP1A were clearly increased in fish caged close to the PW outfall. Induction of plasma vitellogenin was not found in laboratory or field exposures, suggesting that the levels of oestrogen agonists (such as APs) might not have been sufficient to elicit induction, under the present conditions. The applicability of the biomarkers for use in water column biomonitoring programs is discussed.     

Optimization of nutrient component for diesel oil degradation by Rhodococcus erythropolis

A novel bacterium T7-2 was isolated from the oil-polluted sea-bed mud of Bohai Sea, northern China, which can degrade diesel oil at 15 °C. This bacterium was identified as a strain of Rhodococcus erythropolis according to its 16S rDNA gene. In order to enhance degradation efficiency, a five-level, three-factor central composite design was employed to optimize the nutrition supplied to artificial seawater. The results indicate that a supplement of 2.53 g (NH₄)₂SO₄ L⁻¹, 2.75 g Na₂HPO₄ L⁻¹ and 0.01 g yeast extract L⁻¹ to artificial seawater increases the degradation rate from 12.61% to 75% within 7 d.     

Morphological changes in branchial mitochondria-rich cells of the teleost Paralichthys olivaceus as a potential indicator of CO2 impacts

We studied the morphological and biochemical changes of mitochondria-rich cells (MRCs) of a demersal teleost, Paralichthys olivaceus, during exposure to 0.98, 2.97 and 4.95 kPa pCO₂. The apical opening area of MRCs increased 2.2 and 4.1 times by 24 h exposure to 2.97 and 4.95 kPa pCO₂, respectively, while the cross-sectional area or density of MRCs did not change. Gill Na⁺/K⁺-ATPase activity more than doubled at 72 h and then returned to the pre-exposure level at 168 h in 0.98 kPa pCO₂, while it increased 1.7 times at 24 h at 4.95 kPa. These results indicate that the apical opening area of MRCs and the gill Na⁺/K⁺-ATPase activity may be used as an indicator of acute (up to 72 h), but not chronic, impacts of high (>1 kPa) seawater CO₂ conditions in P. olivacues. Limitations of those parameters as indices of CO₂ impacts are discussed.     

A mechanistic study on the photodegradation of Irgarol-1051 in natural seawater

The kinetics of the photoinduced degradation and transformation of the antifouling booster biocide, Irgarol-1051, in natural coastal seawater was studied. The measured first-order rate constant for the degradation of Irgarol-1051 was 4.02 ± 0.1 × 10⁻⁴ h⁻¹, while the rate constant for the formation of 2-methylthio-4-tert-butylamino-6-amino-s-triazine (M1), the most dominant degradation product of Irgarol-1051, was 4.6 ± 0.1 × 10⁻⁵ h⁻¹. This considerably slower rate suggested that the transformation of Irgarol-1051 to M1 may not be the predominant pathway of the photodegradation process. During the photodegradation study, a new s-triazine species was observed in the degradation mixtures which, together with M1, appeared immediately upon photolysis and continued to accumulate in the degradation mixture throughout the entire study duration. This is in contrast to the behaviour of the recently identified degradation product of Irgarol-1051, 3-[4-tert-butylamino-6-methylthiol-s-triazin-2-ylamino]- propionaldehyde (M2), which was only detected in the degradation mixture after a long induction period. High-resolution tandem mass spectrometric analysis hinted that the new degradation product (M4) may possess a terminal alcohol and is likely to be an N-allylic alcohol derivative of M1. This suggests that M4 may, indeed, be a precursor of M2 via redox transformation at its N-allylic alcohol functionality.     

Cytochrome P450 1A1 expression in cetacean skin biopsies from the Indian Ocean

The study describes cytochrome P450 1A1 (CYPA1) expression in the skin of different cetacean species (Megaptera novaeangliae, n = 15; Stenella attenuata, n = 7 and Stenella longirostris, n = 24) from the Mozambique Channel island of Mayotte. Immunohistochemical examination was performed with a monoclonal antibody against scup cytochrome CYPA1. The sex was determined using a molecular approach consisting in the genotyping sex-specific genes. CYPA1 was detected at the junction between epidermis and blubber on dolphins only, mostly in the endothelial cells. Similar observation was obtained in the dermis of one M. novaeangliae. Immunohistochemical slides were scored to evaluate the expression of the CYPA1 and a higher expression was observed in S. longirostris, suggesting a higher exposure to pollutants for this species. The difference of expression between sexes was not significant.     

Effects of chronic exposure to dispersed oil on selected reproductive processes in adult blue mussels (Mytilus edulis) and the consequences for the early life stages of their larvae

Mussels (Mytilus edulis) were continuously exposed to dispersed crude oil (0.015-0.25 mg/l) for 7 months covering the whole gamete development cycle. After 1 month exposure to 0.25 mg oil/l, the level of alkali-labile phosphates (ALP) and the volume density of atretic oocytes in female gonads were higher than those in the gonads of control females, indicating that oil affected the level of vitellogenin-like proteins and gamete development. Spawning of mussels was induced after 7 months oil exposure. Parental oil exposure did not affect subsequent fertilization success in clean seawater but this was reduced in 0.25 mg oil/l. Parental exposure to 0.25 mg oil/l caused both slow development and a higher percentage of abnormalities in D-shell larvae 2 days post-fertilization; reduced growth 7 days post-fertilization. These effects were greatly enhanced when larval stages were maintained at 0.25 mg oil/l. Similar studies are warranted for risk assessment prognosis.     

Impact of elevated levels of CO2 on animal mediated ecosystem function: The modification of sediment nutrient fluxes by burrowing urchins

A mesocosm experiment was conducted to quantify the relationships between the presence and body size of two burrowing heart urchins (Brissopsis lyrifera and Echinocardium cordatum) and rates of sediment nutrient flux. Furthermore, the impact of seawater acidification on these relationships was determined during this 40-day exposure experiment. Using carbon dioxide (CO₂) gas, seawater was acidified to pH_NBS 7.6, 7.2 or 6.8. Control treatments were maintained in natural seawater (pH ≈ 8.0). Under normocapnic conditions, burrowing urchins were seen to reduce the sediment uptake of nitrite or nitrate whilst enhancing the release of silicate and phosphate. In acidified (hypercapnic) treatments, the biological control of biogeochemical cycles by urchins was significantly affected, probably through the combined impacts of high CO₂ on nitrifying bacteria, benthic algae and urchin behaviour. This study highlights the importance of considering biological interactions when predicting the consequences of seawater acidification on ecosystem function.     

Prey and plastic ingestion of Pacific Northern Fulmars (Fulmarus glacialis rogersii) from Monterey Bay,California

Marine plastic pollution affects seabirds, including Pacific Northern Fulmars (Fulmarus glacialis rodgersii), that feed at the surface and mistake plastic for prey or incidentally ingest it. Direct and indirect health issues can result, including satiety and possibly leading to inefficient foraging. Our objective was to examine fulmar body condition, identify cephalopod diet to species, enumerate and weigh ingested plastic, and determine if prey number and size were correlated with ingested plastics in beach-cast fulmars wintering in Monterey Bay California (2003, n = 178: 2007, n = 185). Fulmars consumed mostly Gonatus pyros, G. onyx, and G. californiensis of similar size for both years. We found a significant negative correlation between pectoral muscle index and average size of cephalopod beaks per stomach; a significant increase in plastic categories between 2003 and 2007; and no significant correlation between number and mass of plastic compared with number and size of prey for either year.     

Temporal and spatial biomonitoring of heavy metals in eastern Aegean coastal waters using Amphibalanus amphitrite

This biomonitoring study presents the spatial and temporal distributions of heavy metals in the soft tissues of a major fouling species Amphibalanus amphitrite living on hard substrate at different sites along the eastern Aegean coast. A. amphitrite has been chosen as a strong candidate for monitoring heavy metals. Sediment and seawater samples were also collected to detect their metal contents in order to gain more information on the environmental conditions and possible bioaccumulation patterns. The physico-chemical characteristics of sampling stations have been measured in order to characterize the sampling area. The order of metal concentrations in barnacles, sediment and seawater decreased in the following order Cu > Fe > Zn > Mn > Cd > Cr > Pb > Hg, Fe > Mn > Zn > Cu > Pb > Cr > Hg > Cd and Fe > Zn > Mn > Cu > Pb > Cr > Cd > Hg, respectively. These results showed that barnacles accumulate Cu in a higher degree than both sediment and seawater. Moreover, metal concentrations in barnacle have the potential for use in any future regulatory framework monitoring and eventually controlling ambient metal pollution levels.     

Spatial distribution of polyfluoroalkyl compounds in dab (Limanda limanda) bile fluids from Iceland and the North Sea

The spatial distribution of polyfluoroalkyl compounds (PFCs) was investigated in dab (Limanda limanda) bile fluids collected from Iceland and the North Sea. Concentrations of various PFCs, including perfluorinated sulfonates (C₄-C₆, C₈ PFSAs), perfluorinated carboxylic acids (C₉-C₁₄ PFCAs) and n-methyl perfluorooctane sulfonamidoethanol (MeFOSE), were quantified. Perfluorooctane sulfonate (PFOS) was the predominant compound with highest concentrations along the Danish and German coast (mean 9.36 ng/g wet weight (ww)). Significantly lower PFOS concentrations were found at the other sampling stations in the North Sea and Iceland (p < 0.01, t-test). Conversely, the spatial distribution of the PFCAs in Iceland and the North Sea was more uniform. The most abundant PFCA was perfluorononanoic acid (PFNA), while the mean concentration decreased with increasing chain length from 4.7 ng/g ww for PFNA to 0.04 ng/g ww for perfluorotetradecanoic acid (PFTeDA). Overall, the different spatial distribution of PFCs indicates different origin of sources and different transportation mechanism.     

Acetylcholinesterase,glutathione and hepatosomatic index as potential biomarkers of sewage pollution and depuration in fish

The current study was designed to validate the biomarkers of sewage pollution in Mozambique Tilapia (Tilapia mossambica, Peters) reared in sewage treatment plant (STP) effluent in Ras Al Khaimah, United Arab Emerates, before and following depuration/detoxification. Cellular biomarkers, cholinesterase activity using acetylcholine as a substrate (acetylcholinesterase AChE) and reduced glutathione (GSH) and hepatosomatic index (HSI) were investigated in fresh water fish, Tilapia, raised in a fish farm (Group I/Clean, as Control), treated sewage water/TSW (Group II/Sewage) and thereafter exposed to fresh water in an aquarium for 6 weeks (Group III/Depurated) for depuration. The results showed significantly lower levels of AChE activities in liver (26% p < 0.01) and muscle (30% p < 0.01) of the fish reared in the STP water (Group II/Sewage) as compared to those recorded in the fish from fish farm (Group I/Clean). The depressed AChE level was fully restored in the muscle but partially in the liver after depuration (Group III/Depurated). In contrast, GSH levels were significantly raised in both liver (1.3-fold p < 0.01) and muscle (4-fold) of Group II fish as compared to Group I (control) fish raised in fish farm and following depuration in fresh water (Group III/Depurated) elevated GSH level in liver restored to control values, while remained unchanged in muscle. The average hepatosomatic index (HSI = weight of liver × 100/total fish weight), an indicator of hepatomegaly, in the Group II fish reared in TSW was also significantly higher than that in the reference Group I fish, but decreased to control level in Group III fish following depuration. This study suggests the importance of cellular biomarkers, AChE, GSH and hepatosomatic index in monitoring the impact of sewage water pollution on fish caused by a complex mixture of chemico-biological contaminants and its mitigation following depuration, an effective mean of fish detoxification.     

Trophic interactions of the pelagic ciliate Stentor spp. in North Patagonian lakes

The zooplankton of oligotrophic lakes in North Patagonia is often dominated by mixotrophic ciliates, particularly Stentor amethystinus and Stentor araucanus. Therefore, we tested whether Stentor spp. (i) is an important food for juvenile endemic (Cheirodon australe, Galaxias maculatus, Odontesthes mauleanum, Percichthys trucha) and introduced (Oncorhynchus mykiss) fish species, and (ii) represents a remarkable grazer of bacteria. Ingestion rates of fish estimated by disappearance of Stentor in feeding experiments ranged between 8 (G. maculatus) and 53 (C. australe) ciliates per fish and day, and assimilation rates measured by using radioactively labelled Stentor ranged between 3 (P. trucha) and 52 (C. australe) ciliates per fish and day. However, although we detected the consumption of Stentor by fish, the daily consumption amounted to at most 0.2% of the fish biomass which can not cover the energy requirement of the fish. Furthermore, the daily consumption was equivalent to a maximum of 1.6% of the Stentor standing stock so that fish predation does not seem to be an important mortality factor for the ciliates. The clearance rate of Stentor sp. on natural bacteria was on average 3.8 μl cil⁻¹ h⁻¹. The daily ingestion (mean 3.9 ng C cil⁻¹ d⁻¹) was about 3.5% of the individual biomass of Stentor sp. Therefore, bacteria ingestion might explain a ciliate growth rate of appr. 1% d⁻¹, which was about 17% of the photosynthesis of endosymbiotic algae. The maximum density of Stentor sp. in the lake could ingest about 1 μg C L⁻¹ d⁻¹ bacteria which is only 3% of average bacterial production. Thus, grazing by Stentor sp. does not seem to be a main loss factor for the bacteria.     

The relationships between particulate and soluble alkaline phosphatase activities and the concentration of phosphorus dissolved in the seawater of Toulon Bay (NW Mediterranean)

The activities of particulate and soluble phosphatase were analyzed monthly for 1 year in the coastal ecosystems of the NW Mediterranean Sea. The mean contribution of the particulate activity increased from 56% at an MUF-P concentration of 30 μM to 77% at 0.04 μM. This particulate activity was negatively correlated with the DIP, DOP and TDP concentrations when the activities were related to the seawater volume, chlorophyll a or the protein concentration. The TDP correlations were highly significant (p: 0.001). The DOP correlations were significant (p: 0.04) and became highly significant (p: 0.009) at low DIP concentrations (<0.13 μM). The DIP correlations were significant (p: 0.04) only at low DOP concentrations (<0.18 μM). Thus, the effects of seawater DIP and DOP were found to be linked. The soluble activity exhibited distinct phosphatase fractions with high (0.5–29.5 μM) and low (0.02–2 μM) Km values, but none exhibited significant correlations with phosphorus compounds.