Jarle Klungsøyr

Species characteristics of the hepatic xenobiotic and steroid biotransformation systems of two teleost fish, Atlantic cod (Gadus morhua) and rainbow trout (Salmo gairdneri)

Groups of Atlantic cod and rainbow trout were treated (ip) with beta-naphthoflavone (BNF), phenobarbital, or peanut oil (controls), and properties of the hepatic xenobiotic and steroid metabolizing enzyme systems were evaluated. In both species, BNF treatment resulted in significant induction of microsomal 7-ethoxycoumarin O-deethylase, 7-ethoxyresorufin O-deethylase, biphenyl 4-hydroxylase, and phenanthrene oxidation, especially at the 1,2-position. Immunochemical studies with rabbit IgG prepared against the major BNF-inducible cytochrome P-450 in cod, P-450c, revealed increased amounts of immunoreactive protein in liver slices from both species after BNF treatment. The molecular weight of the induced protein was approximately 58,000 Da, as shown by Western blotting. When titrating biphenyl 4-hydroxylation, however, the antibodies distinguished between the two species, inhibiting the activity of BNF-induced cod 90% and that of rainbow trout 40% at 10 mg IgG/nmol P-450. Furthermore, cytochrome b5 content and UDP-glucuronyltransferase activity were significantly induced only in rainbow trout, whereas the specific content of cytochrome P-450 was significantly increased only in cod. Differences between the two species were observed in the levels of constitutive activities, the amount of induction, and in the regioselectivity of phenanthrene oxidation and androstenedione metabolism. Treatment with phenobarbital showed no effect on any of the parameters investigated in either species. The results show that although there are many common features of the hepatic xenobiotic and steroid biotransformation systems of the two teleosts, certain species characteristics exist in constitutive properties and induction responses.

Environmental impacts of produced water and drilling waste discharges from the Norwegian offshore petroleum industry

Operational discharges of produced water and drill cuttings from offshore oil and gas platforms are a continuous source of contaminants to continental shelf ecosystems. This paper reviews recent research on the biological effects of such discharges with focus on the Norwegian Continental Shelf. The greatest concern is linked to effects of produced water. Alkylphenols (AP) and polyaromatic hydrocarbons (PAH) from produced water accumulate in cod and blue mussel caged near outlets, but are rapidly metabolized in cod. APs, naphtenic acids, and PAHs may disturb reproductive functions, and affect several chemical, biochemical and genetic biomarkers. Toxic concentrations seem restricted to <2 km distance. At the peak of discharge of oil-contaminated cuttings fauna disturbance was found at more than 5 km from some platforms, but is now seldom detected beyond 500 m. Water-based cuttings may seriously affect biomarkers in filter feeding bivalves, and cause elevated sediment oxygen consumption and mortality in benthic fauna. Effects levels occur within 0.5-1 km distance. The stress is mainly physical. The risk of widespread, long term impact from the operational discharges on populations and the ecosystem is presently considered low, but this cannot be verified from the published literature.

Environmental contaminants and biochemical responses in flatfish from the Hvaler Archipelago in Norway.

The levels of several environmental contaminants, including selected polyaromatic hydrocarbons (PAH), organochlorines (DDT/DDE, hexachlorobenzene), 15 polychlorinated biphenyl (PCB) congeners, and polychlorinated dibenzofurans and dibenzo-p-dioxins, PCDF/PCDD), and heavy metals (Cd, Hg, Pb, and As) were analyzed in muscle and liver of three different flatfish species (dab,Limanda limanda; flounder,Platichthys flesus; plaice,Pleuronectes platessa) caught by gill netting at different sites in the Hvaler Archipelago. Indices of biochemical effects in liver S9-fractions were studied by measuring cytochrome P450-dependent monooxygenase and UDP-glucuronyl transferase activities, and by immunoquantitating cytochrome P450 1A1 using an indirect enzyme-linked immunosorbent assay (ELISA). Only low levels of PCDD/PCDF, Cd, and Pb were observed, whereas PCB levels were significantly elevated in fish from the inner sites of the Archipelago compared to a reference site. The contaminant gradient toward the Glomma estuary was correlated with increased cytochrome P450 1A1 activity, measured as 7-ethoxyresorufin O-deethylase (EROD), and with immunoquantitated P450 1A1. In contrast, fish from the site at Idefjorden, although containing elevated contaminant levels, did not show elevated EROD activity, but apparently elevated P450 1A1 protein. These findings may reflect different pollution histories of the sites, and indicate the applicability of biochemical effect indices (i.e., EROD and P450 1A1 immunoquantitation) to monitoring studies. The integrated chemical-biochemical approach employed in this study can obviously be expanded to give fruitful information about cause-effect relationships in other contaminant situations.

Accumulation and effects of aromatic and chlorinated hydrocarbons in juvenile Atlantic cod (Gadus morhua) caged in a polluted fjord (Sørfjorden, Norway)

Abstract Juvenile Atlantic cod were placed in net cages on the bottom at 20–30 m depth at four sites from the inner end of Sorfjorden (Hardanger, Western Norway), to the northern mouth of the fjord in October 1990. After 4 weeks the fish were killed, and liver samples were analyzed for aromatic and chlorinated hydrocarbons (PAHs and PCBs). Liver and gill were also analyzed for cytochrome P450 1A induction, a biomarker for exposure and effects of these contaminants, using catalytic measurements (7-ethoxyresorufin-O-deethylase, EROD) and immunodetection (P450 1A1-ELISA). The caging resulted in the accumulation of a number of PAHs, but very little PCBs, in the liver of the cod, with an inward gradient in the fjord. The gradient, although not dramatic in absolute terms, was parallelled by elevated levels of P450 1A1 when measured by EROD and, partly, by ELISA. The caging strategy seems to be a promising way to approach ecotoxicologically relevant problems, such as bioavailability of contaminants, biomarker responses in the field, and dose-response relationships, also under mixed contaminant situations.

Natural background and anthropogenic inputs of polycyclic aromatic hydrocarbons (PAH) in sediments of South-Western Barents Sea

Polycyclic aromatic hydrocarbons (PAH) were measured in sediment cores from 13 locations in South-Western Barents Sea as part of a detailed study of the Norwegian seabed under the MAREANO program. The generally low PAH levels found, an average around 200 ng g(-1) dry weight for sum PAH, indicate low inputs of petroleum hydrocarbons to the marine environment in the area. Differences in PAH composition and various PAH ratios indicate a natural, mostly petrogenic origin of PAH in sediments from the open sea locations, while the fjord locations show higher pyrogenic PAH contents with an increase towards upper sediment layers, indicating low inputs from human activities. Petrogenic PAH levels increase in deeper sediments at open sea locations, also when normalised to total organic carbon (TOC) contents, suggesting natural leakages of oil-related hydrocarbons in the area.

Development of Atlantic cod (Gadus morhua) exposed to produced water during early life stages: Effects on embryos, larvae, and juvenile fish

Produced water (PW) contains numerous toxic compounds of natural origin, such as dispersed oil, metals, alkylphenols (APs), and polycyclic aromatic hydrocarbons (PAHs). In addition, PW also contains many different chemicals which have been added during the oil production process. In the study described here, cod were exposed to real PW collected from an oil production platform in the North Sea. This was done in order to best recreate the most realistic field-exposure regime in which fish will be affected by a wide range of chemicals. The biological effects found in this study therefore cannot be assigned to one group of chemicals alone, but are the result of exposure to the complex chemical mixture found in real PW. Since APs are well known to cause endocrine disruption in marine organisms, we focused our chemical analysis on APs in an attempt to better understand the long-term effects of APs from PW on the biology of fish. In this study, cod were exposed to several concentrations of real PW and 17β-oestradiol (E(2)), a natural oestrogen, at different developmental stages. Cod were exposed to PW either during the embryo and early larvae stage (up to 3 months of age) or during the early juvenile stage (from 3 to 6 months of age). Results showed that, in general, APs bioconcentrate in fish tissue in a dose and developmental stage dependent manner during PW exposure. However, juveniles appeared able to effectively metabolise the short chain APs. Importantly, PW exposure had no effect on embryo survival or hatching success. However, 1% PW clearly interfered with the development of normal larval pigmentation. After hatching most of the larvae exposed to 1% PW failed to begin feeding and died of starvation. This inability to feed may be linked to the increased incidence of jaw deformities seen in these larvae. In addition, cod exposed to 1% PW, had significantly higher levels of the biomarkers vitellogenin and CYP1A in plasma and liver, respectively. No similar effects were seen in cod exposed to either 0.1% or 0.01% PW.

Biomarkers in Natural Fish Populations Indicate Adverse Biological Effects of Offshore Oil Production

Background Despite the growing awareness of the necessity of a sustainable development, the global economy continues to depend largely on the consumption of non-renewable energy resources. One such energy resource is fossil oil extracted from the seabed at offshore oil platforms. This type of oil production causes continuous environmental pollution from drilling waste, discharge of large amounts of produced water, and accidental spills. Methods and principal findings Samples from natural populations of haddock (Melanogrammus aeglefinus) and Atlantic cod (Gadus morhua) in two North Sea areas with extensive oil production were investigated. Exposure to and uptake of polycyclic aromatic hydrocarbons (PAHs) were demonstrated, and biomarker analyses revealed adverse biological effects, including induction of biotransformation enzymes, oxidative stress, altered fatty acid composition, and genotoxicity. Genotoxicity was reflected by a hepatic DNA adduct pattern typical for exposure to a mixture of PAHs. Control material was collected from a North Sea area without oil production and from remote Icelandic waters. The difference between the two control areas indicates significant background pollution in the North Sea. Conclusion It is most remarkable to obtain biomarker responses in natural fish populations in the open sea that are similar to the biomarker responses in fish from highly polluted areas close to a point source. Risk assessment of various threats to the marine fish populations in the North Sea, such as overfishing, global warming, and eutrophication, should also take into account the ecologically relevant impact of offshore oil production.

May organic pollutants affect fish populations in the North Sea

The North Sea is a highly productive area with large fish populations that have been extensively harvested over the past century. North Sea fisheries remain important to the surrounding countries despite declining fish stocks over the past decades. The main reason for declining fish stocks is nearly certainly overfishing, but other environmental pressures also affect fish populations, such as eutrophication, climate change, and exposure to metals and organic pollutants, including polyaromatic hydrocat bons (PAHs), alkylphenols, and organochlorine compounds. There are three main sources of organic pollutants in the North Sea: atmospheric, land-based sources, and inputs from offshore gas and oil installations. All three sources contribute to elevated concentrations of organic pollutants in the North Sea compared to the Norwegian Sea. There is evidence that chlorinated organic contaminants were present in sufficiently high concentrations in the southern North Sea two decades ago, to alter embryonal development in fish. The results from extensive, long-term monitoring programs show that some diseases decreased whereas other increased in the southern North Sea and that, among other factors, contaminants may play a role in the temporal changes recorded in disease prevalence. Recent studies demonstrated that components in offshore effluents may affect fish reproduction and that tissues of fish near oil rigs are structurally different to tissues of fish from reference areas. Data on effluents from offshore activities have recently become available through an international workshop (BECPELAG) and follow-up studies.

Initial characterization of the hepatic microsomal cytochrome P-450-system of the piked whale (Minke) Balaenoptera acutorostrata

Abstract Components of the microsomal cytochrome P -450- system were measured in liver samples from several female, one male and two foetal piked whales. In the same samples, four eytochrome P -450- dependent monooxygenase activities were assayed, protein pattern was determined by SDS-polyacrylamide gel e electrophoresis and the regiospecificity of phenanthrene metabolism in vitro was determined by gas chromatography/mass spectrometry. Foetal samples contained cytochrome P -450, cytochrome b 5 and NADPH-cytochrome P -450,reductase, but at lower levels than adults. Of the monooxygenase activities, only the O -deethylation reactions (7-EROD ∗ and 7-ECOD) could be detected in the foetuses. The turnover of 7-ethoxyresorufin was even higher than in most ofthe adult samples. In the adult samples, apparent differences could be observed in the parameters investigated between immature specimens of different sex and between immature and mature female whales. In vitro metabolism of phenanthrene showed a clear preference for attack at the 9,10-position by adult liver microsomes, where 10–35% appeared as the 1,2-dihydrodiol, with large individual variations. When metabolized by foetal liver microsomes, only 25–35% of total phenanthrene metabolites appeared as the 9,10-dihydrodiol, whereas 38–50% were the 1,2-dihydrodiol. Between 16 and 32% of the metabolites in all samples were phenols.

PAH metabolites in bile and EROD activity in North Sea fish

Abstract Atlantic cod ( Gadus morhua ), haddock ( Melanogrammus aeglefinus ), and long rough dab ( Hippoglossoides platessoides ) were sampled from areas with oil production in the North Sea. Egersundbanken, an area without oil production, was used as a reference site. Ethoxyresorufin- O -deethylase (EROD) activity was measured in liver to estimate cytochrome P4501A levels. Bile was analysed by fixed wavelength fluorescence (FF) detection for polyaromatic hydrocarbon (PAH) metabolites. No increased levels of EROD activity were detected in fish caught in the vicinity of oil production fields, when compared to the reference site, Egersundbanken. No differences in levels of PAH metabolites in bile were detected between the stations for any of the species. Therefore, contamination from 2–5 ring PAH compounds in the vicinity of oil fields in the North Sea may not be of great concern for these fish species.