Langtidsvirkninger av utslipp til sjø fra
petroleumsvirksomheten (PROOF)
årsmøte 2005
Solstrand Fjord Hotell, 27. – 29. september 2005
Sammendrag fra prosjektene
Innholdsfortegnelse
TEMA: ARKTIS .............................................................................................................. 4
Leonel Camus/Thierry Bassant: long term effects on Arctic ecosystem from accidental
discharges .............................................................................................................................................. 4
Odd Gunnar Brakstad: Interactions of microbes with oil-polluted Arctic fjord ice .......................... 4
Guri Faksness: Dissolution of Water Soluble Compounds from Oil Encapsulated in Ice ............... 5
Per Johan Brandvik: Fate and behavior of oil spills under Arctic conditions Earlier results
compared with new field experiments on Svalbard ............................................................................ 6
TEMA: EFFEKTER AV PRODUSERT VANN PÅ TORSK ......................................................... 7
Stephan Boitsov: Identification of Estrogen-Like Compounds in Produced Water from Offshore
Oil Installations. ..................................................................................................................................... 7
Grete Jonsson: Analyse av alkylfenol metabolitter I fiskegalla; metodeutvikling og anvendelse .. 7
Jarle Klungsøyr: Effects on development, sex differentiation and reproduction of cod (Gadus
morhua) exposed to produced water during early life stages ........................................................... 8
Sonnich Meier: Effect on the timing of spawning, sperm quality and the realized fecundity of cod
(Gadus morhua) after a long-term exposure to produced water and alkylphenols.......................... 8
Rajdeep Sidhu: Radioaktivitet i produsert vann fra norske olje- og gass installasjoner –
konsentrasjoner, biotilgjengelighet og doser til mennesker og marin biota .................................... 8
TEMA: UNRESOLVED COMPLEX MIXTURE – UCM .............................................................. 9
Alf Melbye: Chemical Characterisation of Polar Components in Produced Water ......................... 9
Trond Nordtug: : Effects of the unresolved complex mixture (UCM) of petrogenic oils: Impacts in
the seawater column............................................................................................................................ 10
Bodil Larsen: Overlevelse av rekelarver (Pandalus borealis) unner eksponering til vannløselige
olje komponenter og lys: kopling til biomarkører. ............................................................................ 10
TEMA: BIOMARKØRER/RISK ......................................................................................... 11
Kevin Thomas: Bioassay-assisted monitoring of produced water effluents and environmental
samples from the Norwegian sector of North Sea. ........................................................................... 11
Merete Grung : Use of Passive sampling devices in monitoring of potential impact of offshore
discharges and accidental oil spills (PASSIMPACT) ........................................................................ 11
Tor Fredrik Holth: Predicting chronic effects in fish from sublethal markers (PredFish) ............. 12
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Steinar Sanni: ....................................................................................................................................... 13
Arnfinn Skadsheim: DEEP-SEA: knowledge status and methods for impact and risk assessment
............................................................................................................................................................... 13
Odd Ketil Andersen: Proteome Expression Signatures (PES) in fish as a diagnostic tool to
evaluate the environmental impacts of offshore oil and gas exploration ....................................... 14
Bjørn Einar Grøsvik: Effects of oil and produced water on atlantic cod; biomarker development
with proteomics based methods ........................................................................................................ 14
TEMA: MODELERING.................................................................................................... 15
Anne Solberg:....................................................................................................................................... 15
Anders Bjørgesæter: THE DEVELOPMENT OF FIELD-BASED SPECIES SENSITIVITY
DISTRIBUTIONS (F-SSDs). .................................................................................................................. 15
Henrik Rye: Utvikling av sediment modell for simulering av miljøeffekter som følge av utslipp av
borekaks og boreslam. ........................................................................................................................ 15
Frode Olsgard: Parameterisation of the Environmental Impacts on Bottom Fauna of Water-based
Drilling Fluids and Cuttings – Field and Mesocosm Experiments (PEIOFF-FAME) ....................... 16
Renee Katrin Bechmann: Effekter av boreslam på torsk og skjell .................................................. 16
JoLynn Carroll: Environmental effects of offshore oil activities: Experimental tests of petroleumassociated components on benthos at community, individual, and cellular levels....................... 17
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TEMA: Arktis
Leonel Camus/Thierry Bassant: long term effects on Arctic ecosystem from
accidental discharges
Project: PROOF No 164407
T. Baussant*, A. Skadsheim, B.K. Larsen and S. Sanni - RF-Akvamiljø, Mekjarvik 12,
4070 Randaberg
L. Camus* - Akvaplan-niva, polar Environmental center, 9296 Tromsø
Jørgen Berge – The University Centre on Svalbard, PB 156, 9170 Longyearbyen
R. Krapp - University of Kiel - Institute for Polar Ecology, 24148 Kiel, Germany
*presenting authors
Summary – Oil and gas activities are moving in Arctic pristine areas. This requires
investigations to assess their possible environmental impact. The main objective of this
project is to investigate possible long term effects of oil drifting to Arctic sea ice fauna
from accidental discharges and offshore installations. A marked seasonality with regard
to ice cover and illumination, and the constant low temperature feature the Arctic
environment. Also short trophic chains rely on some key species like the crustacean
amphipod Gammarus wilkitzkii, a species exclusively found in ice fauna. In case of spill
to the ice margin, several routes and scenarios of exposure to the amphipod are relevant.
In this project, experiments will be performed to simulate exposures via water and food
following the weathering of oil compounds entrapped in ice during spring ice break-up.
In G. wilkitzkii, the uptake of the bioavailable fraction will be measured. Short- to
medium-term biological responses (biomarkers), after continuous or repeated exposures
and with intermittent restitution periods, will be analysed as a way to assess longer term
effects of oil. Also, a link with ecologically-relevant impacts associated to the
reproduction cycle of the amphipod will be established. The aspects related to UV
irradiance in ice-covered water will be examined by a PhD study connected to the
project and also, through a close collaboration with another ongoing NFR project
(“phototoxicity study”; NFR° 159176). A main experiment is planned in the autumn
2005 after collection of a sufficient number of G. wilkitzii around Spitzbergen. This
presentation will describe the main subtasks assigned to the project and their
participants, the exposure setup and analytical methodologies. Also, the results from a
short uptake study with G. wilkitzkii exposed to oil in water (WAF) and oil in ice will be
shown.
Odd Gunnar Brakstad: Interactions of microbes with oil-polluted Arctic fjord ice
O.G. Brakstad,1 I. Nonstad,2 Faksness, L.-G.,3 and P.J. Brandvik3
SINTEF Materials and Chemistry, Trondheim1 NTNU, Trondheim2, and UNIS, Longyearbyen3
Oil pollutions caused by production or transport accidents may be transported to the ice margins
of the Arctic oceans by prevailing seawater currents, where the oil becomes infested in the ice. It is well
4
known that the Polar ice harbour high numbers of microbes of diverse community structures, and recent
studies have shown that ice bacteria have metabolic activities at temperatures well below freezing point.
In the PROOF study “Weathering of oilspills under Arctic conditions” we have investigated the
interactions between microbial communities and oil in Arctic fjord ice during a winter period. Crude oils
(Statfjord, Goliat, Heidrun Åre and IF180) were frozen in Fjord ice (Van Mijen fjord, Spitzbergen) in
February 2004, and microbial communities were investigated during a period of 112 days (February 11
to June 2). During this period oil-containing ice cores and references (clean ice) were sampled in March,
April and at the end of the trial. Total numbers of microbes were enumerated in ice core sections by
epifluorescence microscopy, while viable heterotrophic and oil-degrading prokaryotes were quantified by
culture techniques. Changes in microbial communities during the experiment were characterized after
polymerase chain reaction (PCR) amplification of 16S rRNA genes by denaturing gradient gel
electrophoreses (DGGE) and restriction enzyme analysis. Individual members of the communities were
identified by cloning and sequence analysis of 16S rRNA genes. Oil analyses were performed by GCFID and GCMS analysis.
Epifluorescence analysis, in which microbial concentrations in oil-polluted and clean ice were
compared, showed that the microbial numbers increased significantly in core sections associated with
the highest oil contents in April and June samples. These results were confirmed with heterotrophic
prokaryotes and partly with oil-degrading prokaryotes. DGGE analysis revealed that bacterial banding
patterns were reduced in oil-polluted ice during the test period, and a few strong bands seemed to
predominate. In contrast, the numbers of DGGE bands seemed were higher in comparable samples
from clean ice. This indicated that bacterial genotypes were reduced in polluted ice, with a few types
being abundant in these samples. Cloning, restriction enzyme analysis and sequence analysis were
performed on samples from clean ice and ice polluted with Statfjord oil. These results showed that
restriction enzyme fragment patterns were reduced in oil-polluted ice cores when compared to patterns
in clean ice, in agreement with DGGE analysis. Sequence analysis showed that a bacterial type related
to the polar gammaproteobacterium Colwellia sp. were predominant in oil-polluted ice, while a number
of other well-known Polar bacteria were present in clean ice.
In conclusion, oil pollution had a significant impact on the microbial communities in Arctic fjord
ice. Oil pollution resulted in elevated concentrations of prokaryotes, and bacterial diversity was reduced,
with predominance of a few bacterial types known to be involved in oil biodegradation in Arctic
seawater. These results indicate that ice bacteria may degrade oil compounds in Arctic ice at
temperatures well below freezing points. This is new information, and it will be important to investigate
hydrocarbon biodegradation processes and rates for oils frozen in ice, since oil compounds may be
eradicated from the Arctic environment even in the cold winter periods.
Guri Faksness: Dissolution of Water Soluble Compounds from Oil Encapsulated in Ice
Liv-Guri Faksness and Per Johan Brandvik
The University Centre in Svalbard (UNIS), Longyearbyen, Norway
Liv-Guri.Faksness@unis.no
Field work with oil encapsulated in ice was performed to study the dissolution of water soluble
compounds. Six oil types were frozen into fjord ice early in February, 2004. Ice cores with oil were
sampled every third week until June. Oil/ice samples were analyzed and migration of water soluble
compounds from the oil into the ice was quantified. In addition, water accommodated fractions of the
same oils in seawater were generated in the laboratory according to a procedure established by
CROSERF.
The distribution and concentration of water soluble compounds in the field samples were
compared with data for the same oil types obtained in the laboratory experiments. The distribution of
5
compounds in the field was similar as observed in the laboratory, but the field concentrations were
lower.
The results from the field experiments show that the concentration of the water soluble
compounds in the ice is decreasing with ice depth. A concentration gradient as a function of time was
also observed, indicating that there has been a migration of water soluble compounds through brine
channels in the ice to the underlying water. The concentration of water soluble compounds in the bottom
20 cm ice core was reduced from 30 ppb to 6 ppb in the experimental period. Although the
concentrations were low, the exposure time was long (nearly four months). This might indicate that the
ice fauna could be exposed to a substantial dose of toxic water soluble compounds.
Additional field experiments were performed with oil encapsulated in and under ice this winter, and
preliminary results might be included in the presentation.
Per Johan Brandvik: Fate and behavior of oil spills under Arctic conditions
Earlier results compared with new field experiments on Svalbard
P. J. Brandvik1* and L-G Faksness1
University centre in Svalbard (UNIS),
Pb. 156, N-9171 Longyearbyen, Norway
1The
The knowledge regarding weathering processes in Arctic oil spills and especially with ice is
limited. Experimental studies have been performed in laboratories, but only to a limited degree in the
field. This paper summarizes and compares results from field experiments performed in Norway in
1989, 1993 and 2005.
Field measurements from experimental oil releases in Norway are used to compare the behavior
of oil spilled in open water and in an Arctic broken ice scenario. Similar oil types and amount (25 - 30
m3) were used in an experimental oil release in open water at Haltenbanken (65°N) in 1989 and in
dynamic broken ice at Svalbard (75°N) in 1993. Experiments performed during spring 2005 on Svalbard
are also presented. Several weathering properties for the oil spill in broken ice are strongly influenced
by the low temperature, reduced oil spreading and reduced wave action caused by the high ice
coverage. Reduced water uptake, viscosity, evaporation and pour point extend the operational time
window for several contingency methods compared to oil spills in open waters. This could open up for
dispersant treatment and in-situ burning even after an extended period of weathering for an oil spill in
broken ice.
In 2003 SINTEF and the University Centre in Svalbard (UNIS) and co-workers
(SINTEF/NILU/UiB) initiated a three year research program as a part of the PROOF program to study
selected weathering processes in Arctic oil spills for a wide range of oil types. This three-year program if
funded by Norwegian authorities and oil companies. The objective with this program is to focus on the
weathering processes; biodegradation, release of water-soluble components to the seawater and photo
oxidation. Results from field experiments to study oil behaviour (evaporation, emulsification, spreading
etc.) with different ice conditions (slush ice, 30% and 90% ice coverage) are presented in this paper.
Keywords: oil spill; weathering; Arctic; ice; field experiments.
6
TEMA: Effekter av produsert vann på torsk
Stephan Boitsov: Identification of Estrogen-Like Compounds in Produced Water from Offshore
Oil Installations.
Stepan Boitsov* and Sonnich Meier, Institute of Marine Research (IMR)
Svein Mjøs, Norwegian Institute of Fisheries and Aquaculture Research (NIFAR)
Jon Sognstad, Department of Chemistry, University of Bergen
*Institute of Marine Research, Post Box 1870 Nordnes, N-5817 Bergen, Norway
Fax: +47-55238584, e-mail address: stepan.boitsov@imr.no
Significant quantities of alkylphenols are released into the sea from oil installations as a result
of produced water discharges. It has been shown that alkylphenols have estrogenic effects on marine
biota. The alkylphenol fraction of produced water is most likely not acutely toxic due to its rapid dilution
upon release into the sea. However, chronic effects may occur over time due to bioaccumulation.
Methods developed at IMR allow the determination of alkylphenols of various alkyl chain lengths in
produced water and fish tissues.
However, the above-mentioned methods have been validated for 23 alkylphenols only. A large number
of various isomers of C0-C9 alkylphenols is found in an average produced water sample. Most of these
have not been identified due to absence of standards. At the same time, it is only some of the
alkylphenol isomers that have been shown to exert a significant toxic effect on marine biota. Thus, longchain, branched para-substituted alkylphenols have the strongest estrogenic effect. Therefore, a new
project has been started at IMR with the aim of identifying the potentially estrogenic alkylphenol isomers
present in produced water. Large volumes of produced water from 2 oil platforms have been extracted,
purified by GPC and normal-phase HPLC and analysed by various GC-MS techniques. Standards for
the isomers of interest have been synthesised using suitable procedures. By comparing the GC-MS
data obtained for the produced water samples with that for the synthesised standards, as well as by
using statistical methods (retention indices) and new mathematical techniques (curve resolution), the
alkylphenol isomers of interest are analysed. Certain other toxic components of produced water, such
as thiols, are also analysed .
Grete Jonsson: Analyse av alkylfenol metabolitter I fiskegalla; metodeutvikling og
anvendelse
Forfattere: Grete Jonsson, Admira Cavcic, Tone Stokke, Kjellaug Aarebrot, Jonny
Beyer og Kåre Jørgensen
Prosjektet “Pollutant exposure and effects in fish related to the discharge of produced
water in the North Sea oil industry” er inndelt i åtte ulike aktiviteter, hvorav utvikling
av analysemetode for å kunne måle metabolitter av alkylfenoler i fiskegalle er en.
Eksterne samarbeidspartnere er; NIVA – Oslo, ITM - Universitetet i Stockholm, NTNU
- Trondheim og Universitetet i Florence - Italia. Alkylfenoler (APer) finnes i relativt
høye konsentrasjoner i produsert vann, og noen av disse forbindelsene kan være giftige
eller ha hormonforstyrrende egenskaper. På grunn av disse potensielle negative
effektene er det ønskelig å kunne måle en eventuell eksponering av fisk.
I første del av metodeutviklingen ble torsk eksponert for ni ulike alkylfenoler som er
representative for hva som finnes i produsert vann. Væskekromatografi med fluorescens
detektor (HPLC-F) ble så anvendt for å karakterisere alkylfenolmetabolittene som
skilles ut i fiskegallen. For å kunne kvantifisere spesifikke metabolitter i galle fra fisk
7
eksponert for mer komplekse løsninger ble gallen først renset og deretter derivatisert
vha fast fase ekstraksjons kolonner (Solid Phase Analytical Derivatization, SPAD). To
typer derivater ble laget, trimetylsilyl (TMS) og pentaflourobensyl (PFB), og analysert
vha gasskromatografi med massespekter som detektor (GCMS, EI og NCI mode).
Begge GCMS metodene har blitt anvendt for å analysere alkylfenolmetabolitter i galle
fra både laboratorie- og felteksponert fisk. Resultater fra analysene vil bli presentert.
Jarle Klungsøyr: Effects on development, sex differentiation and reproduction of
cod (Gadus morhua) exposed to produced water during early life stages
Sonnich Meier: Effect on the timing of spawning, sperm quality and the realized
fecundity of cod (Gadus morhua) after a long-term exposure to produced water
and alkylphenols.
Authors:
Sonnich Meier; Marianne Petersen; Marita Larsen; Audery Geffen; Stepan Boitsov; Jarle Klungsøyr
Two years old North-Atlantic cod have been orally exposed to different concentrations of a mixture of
alkylphenols (body burden 0,4 – 2000 ppb ≈ 0,8-4000 ng/l) and produced water from oil installation
(Oseberg C, Hydro) (diluted 1:1000 and 1:10000) and a positive control (100 ppb E2) for 20 weeks from
the start of October to the end of February. A high number of fish (30 in each group) were sampled for a
study of a board spectrum of biomarkers and physiological effects. Two parallels of 2 females and 2
males from each treatment were transferred to 2 m3 tanks and were allowed to spawn naturally. Egg
was collected throughout the whole season, and egg quality and hatching success were measured. In
addition, there were performed experiments on exposure (produced water) of embryos from the different
groups to produced water to look for eventual second generation effects.
We will present here the results of the spawning experiment, measurement of sperm quality and the
second generation experiment.
Rajdeep Sidhu: Radioaktivitet i produsert vann fra norske olje- og gass
installasjoner – konsentrasjoner, biotilgjengelighet og doser til mennesker og
marin biota
R. Sidhu, D. Eriksen, E. Strålberg, K. I. Iden (IFE), H. Rye (SINTEF), K. Hylland, A. Ruus (NIVA) og M
Berntssen (NIFES)
Produsert vann fra olje- og gassproduksjon inneholder forhøyde nivåer av naturlig forekommende
radionuklider, i hovedsak 226Ra and 228Ra. I dette prosjektet studeres konsekvensene av utslipp av
radionuklider til det marine miljøet . Hovedhensikten med prosjektet er å etablere radiologisk sikre
8
utslippsgrenser for radium, bly og polonium assosiert med andre komponenter i produsert vann fra oljeog gassinstallasjoner på den norske kontinentalsokkelen. Prosjektet er delt inn i seks delprosjekter:
1. Kartleggig av det naturlige bakgrunnsnivået til de relevante radionuklidene (226Ra, 228Ra) i sjøvann,
sediment og relevante organismer i Nordsjøen.
2. Studier av mobilitet og sedimenteringsmekanismer til de relevante radionuklidene gitt ulike
utslippsscenarier.
3. Studier av biotilgjengelighet av radium i sediment, porevann og vann gitt de samme
utslippsscenarier som i delprosjektet 2.
4. Studier av biologiske effekter av ioniserende stråling på marine organismer. Det skal utføres
eksperimenter med både in vitro- og helkroppseksponering.
5. Modellering av utslipp av produsert vann inneholdende de relevante radionuklidene i sjøvann
utføres med DREAM-modellen utviklet av SINTEF. Både transport, sedimentering og opptak i
relevant biota skal inkluderes.
6. Beregning av doser til individer, befolkningsgrupper og marin biota fra utslipp av produsert vann.
Basert på beregnede doser til marin biota, vil PNEC-verdier (Potential No Effect Concentrations) bli
anbefalt.
Foreløpige resultater og planlagt arbeid vil bli presentert.
TEMA: Unresolved complex mixture – UCM
Alf Melbye: Chemical Characterisation of Polar Components in Produced Water
Jorunn Nerbø Hokstad, Alf Melbye
The main objective of this project is to obtain a thorough understanding of the chemical characteristics
of polar components present in produced water and crude oils. The results from the project will form an
essential basis for evaluation of possible long-term effects of produced water constituents in the marine
environment, and this project cooperates in this matter with the PROOF supported project “The
unresolved complex mixture (UCM) of petrogenic oils: Impacts in the seawater column (Odd Gunnar
Brakstad, SINTEF). The UCM part of the water extract of a crude oil, where all compounds are
bioavailable, has been associated with toxicity to marine organisms. The project concentrates on
fractionating the water soluble fraction of the reservoir biodegraded oil Troll crude which is dominated by
the UCM fraction, and the reference oil Statfjord crude which is a light oil with a quantitatively small
UCM fraction. The first activities in the project have focused on establishing fractionation techniques. A
preparative technique has been established for fractionating the water soluble fraction of crude oil into
14 different fractions. Present methodology for chemical analysis of organic constituents of produced
water focus mainly on hydrocarbons (PAH) and to some extent phenols and naphtenic acids. The
unresolved complex material (UCM) is poorly understood and not considered in regular analysis of
produced water. UCM is known to include polar components, like nitrogen-, sulphur- and oxygencompounds (N,S,O-compounds). Analysis of the different fractions has been performed using GC/MS
and LC/MS techniques. This work has confirmed the complexity of the UCM fraction, but has also
identified different compound groups and general chemical structures. Preliminary results will be
presented.
9
Trond Nordtug: : Effects of the unresolved complex mixture (UCM) of petrogenic
oils: Impacts in the seawater column
T. Nordtug,1 O.G. Brakstad1, A.G. Melbye,1 J. N. Hokstad,1 D. Altin,2 K.-E. Tollefsen3, M. Grung3, D. and
S. Rowland4
SINTEF Materials and Chemistry, Trondheim1, Biotrix, Trondheim2, NIVA, Oslo3, and University of
Plymouth4
The unresolved complex mixtures (UCMs) of petrogenic oils is a recalcitrant fraction of the oil
after discharges to the marine water column. The UCM fraction contain a high proportion of N-,S, and
O-containing compounds, which are polar and therefore represented in the water-soluble fractions
(WSFs) of in-reservoir biodegraded oils. UCM compounds may resist both evaporative and
biodegradation processes in the water column, and photo-transformation processes may result in toxic
UCM-related compounds.
We have investigated the toxicity of this complex mixture in an UCM-rich oil (Troll) and an UCMpoor reference oil (Statfjord). WSFs were generated from oils evaporated at 200°C to remove
monoaromatics and other low boiling point hydrocarbons. The WSFs were separated in a number of
fractions (n=15) by a large scale normal-phase high-pressure liquid chromatography (HPLC) method.
The UCM, observed as an unresolved chromatographic “hump”, was distributed in 4-5 of the fractions.
All fractions were subjected to a solvent exchange to a low-tox solvent (DMSO) and diluted in water
below the toxicity threshold of this solvent. All fractions were then tested with a number of rapid
screening bioassays to compare the toxicities of different UCM-fractions. The results of this bioassayassisted fractionation procedure are under processing and the results presented. Similar studies will be
performed with photo-oxidised oils, since this process may contribute to the UCM-related toxicity. The
sub-acute effects of selected UCM-fractions on marine phytoplankton photosystem II will also be
examined. Toxicity-related data will be available for exposure model tools (Oscar, Dream), either for the
whole UCM-fraction representing one pseudo-oil compound group, or separated in several toxic
fractions partially characterized by chemical analysis.
Bodil Larsen: Overlevelse av rekelarver (Pandalus borealis) unner eksponering til
vannløselige olje komponenter og lys: kopling til biomarkører.
Forfattere: Bodil K. Larsen, Daniela M. Pampanin, Shaw Bamber, Francesco Regoli og Thierry
Baussant
Formålet med prosjektet ” Effect of off-shore oil industry related discharges in the Arctic” er at
undersøke effekter av olje når dyrene samtidig belyses med UV-lys og synlig lys. Olje/tjærestoffer kan
være langt mer giftige i kombinasjon med UV-lys og i arktiske strøk, med 24 timers dagslys, er dette
viktig at ta i betraktning. Nyklekkede rekelarver blev eksponert for 2 uker til to forskjellige
konsentrasjoner av vannløselige oljekomponenter og til to forskjellige lys scenarier: naturlig nivå av
UV/synlig lys eller synlig lys alene. I tillegg blev en gruppe rekelarver eksponert til UV eksponert olje
mens dyrene selv blev belyst med synlig lys. Resultatene bekreftede at UV lys forøker giftigheten av olje
komponenter. Etter 5 dages eksponering sås en signifikant høyere dødelighet hos larver eksponert til
UV og den høyeste konsentrasjon av olje komponenter. Forskjellene mellom grupper blev mer markant
med tid. Hos larver eksponert til UV belyst olje uten selv at blive UV belyst sås en plutselig og kraftig
forøkning i dødelighet i de siste dager før avslutningen av forsøket. Denne markante forøkning i
dødelighet kan henge sammen med at larvene på dette tidspunkt skiftede stadie/skall, og herunder kan
celleveggene muligvis være mer gjennom trengelige for giftige komponenter. Ved avslutningen av
forsøket blev rekelarver frosset ned til forskjellige biomarkør målinger. Resultater av disse vil blive
diskutert. Et preliminært forsøk med torskelarver blev utført, men må gjentas i 2006 da dødeligheten var
for høy i alle grupper. Det er forventet at utføre forsøk med både polartorsk og is amfipoder i etteråret
2005.
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TEMA: Biomarkører/RISK
Kevin Thomas: Bioassay-assisted monitoring of produced water effluents and
environmental samples from the Norwegian sector of North Sea.
Knut-Erik Tollefsen (NIVA), Jan Balaam (CEFAS), Marc Berntssen (NIFES), Christian Dye (NILU),
Eivind Farmen Finne (NIVA), Merete Grung (NIVA), Ketil Hylland (NIVA), Torsten Källquist (NIVA), Leif
Norrgren (SLU), Hege Stubberud (Jordforsk) and Kevin V. Thomas (NIVA)
Oil production platforms discharge large volumes of produced water (PW) containing various chemicals
that may pose a threat to organisms living in the recipient. This applies in particular to groups of
chemicals such as alkylphenols (APs) and polycyclic aromatic hydrocarbons (PAHs) that are suspected
of producing various toxic effects including reproductive disturbances, mutagenicity and carcinogenicity.
Although the toxicity to marine animals are well documented for many of the chemicals that are
found in PW effluents, complex samples such as PW contain a variety of chemicals whose identity and
toxicity is poorly characterised. In the NFR-proof funded pilot project “Identification of ecologically
relevant toxic components in effluents from offshore activities (OffTIE)”, a bioassay-directed
fractionation and chemical identification procedure revealed that PW contain acute toxic chemicals,
CYP1A inducers, oxidative stressors, estrogen mimics and mutagenic chemicals. Some of these groups
of chemicals exhibited properties that may potentially cause accumulation and long-term (ecological)
effects on organisms living in the vicinity of oil/gas production platforms.
The objective of the NFR-proof project no. 159113 “Integrating monitoring methods for impacts
of offshore discharges to the North Sea (IMONIT)” is to develop and deploy a suite of small scale
bioassays for toxicity characterisation of PW effluents and effluent receiving waters in the vicinity of
selected oil/gas production platforms. The combination of bioassay development, validation of different
sampling methodologies and the use of bioassay-directed fractionation and chemical identification
procedures aims to assess; 1) the spatial and temporal variation in PW toxicity, 2) to characterise the
toxicity of the water in the vicinity of oil/gas production platforms and 3) to identify the most toxic
chemicals found in these waters. This presentation will focus on the validation of the suite of bioassays
developed, results from spatial and temporal monitoring of PW effluents from selected production
platforms in the North Sea, and toxicity screening of samples obtained after deployment of passive
sampling devices during the water column survey 2004 at Statfjord B.
Affiliations
NIVA-Norwegian Institute for Water Research (NO), NIFES - Norwegian Institute for Fisheries and
Seafood Research (NO), NILU - Norwegian Institute for Air Research (NO), SLU - Swedish Agricultural
University (SE), Jordforsk (NO), and CEFAS - The Centre for Environment, Fisheries & Aquaculture
Science (UK).
Merete Grung : Use of Passive sampling devices in monitoring of potential impact
of offshore discharges and accidental oil spills (PASSIMPACT)
Merete Grung1, Christopher P. Harman1, Knut-Erik Tollefsen1, Oddvar Røyset1, Ketil Hylland1,3, Alf
Melbye2, Pål Molander3, David Alvarez4 and Graham Mills5
1) NIVA – Norwegian Institute for Water Research – NO, 2) SINTEF – NO, 3) University of Oslo – NO,
4) USGS – U.S. Geological Survey – USA, 5) University of Portsmouth – UK
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The major discharge from offshore activities is produced water (PW). In the Norwegian sector we expect
a rise in the volume of discharge in the years to come due to the increased amounts of produced water
associated with ageing wells. The substances discharged are readily diluted by receiving waters, with
low concentrations predicted to be present a short distance from the point of discharge. Establishing the
occurrence of substances from these discharges in the surrounding environment thus requires sensitive
analytical techniques and pre-concentration processes. Several pre-concentration techniques have
been used in earlier studies to allow for chemical characterisation and bioassay testing. Some of them
represent the analyte concentration at a specific moment of time; others can provide time integrated
information about the target compound concentration in the environment. Knowledge of
uptake/sampling rates for the different target compounds in the passive samplers makes it possible to
estimate the concentrations of these compounds in water. The use of passive samplers may thus
greatly improve the techniques available for monitoring the occurrence of PW specific substances in the
water column. These data are required to ensure the robust assessment of the environmental risk
posed by substances originating from offshore discharges.
The objective of this project is to optimise and validate a suite of passive sampling devices for the
chemical and toxicological screening of organic chemicals from oil related discharges. The sampling
kinetics of the passive sampling devices will be characterised and compared to the sampling kinetics of
blue mussels; a typical sentinel monitoring species. To determine the susceptibility of passive sampling
devices to variations in environmental conditions we will investigate the effects of bio-fouling on passive
sampler sampling kinetics. The approach will be validated using a suite of passive sampling devices and
blue mussels in a field study and simulated spill situation. The results obtained in the field studies will
then be compared to those obtained from the DREAM model using a selection of PW specific
substances.
Tor Fredrik Holth: Predicting chronic effects in fish from sublethal markers
(PredFish)
Holth TF1,2, Grung M1, Rindal Jacobsen M2, Balk L3, Petersen G4, Bernssen M5, Olsvik P5, Camus
L6, Lam P7, Norrgren L8, de Coen W9, Förlin L10, Klobucar G11, Ropstad E12, Aleström P12, Hylland
K1,2
1Norwegian Institute for Water Research (NIVA), Oslo, Norway; 2University of Oslo, Oslo, Norway;
3Stockholm University, Stockholm, Sweden; 4DHI Water & Environment, Hørsholm, Denmark; 5National
Institute of Nutrition and Seafood Research (NIFES), Bergen, Norway; 6Akvaplan-NIVA, Tromsø,
Norway; 7City University of Hong Kong, Hong Kong; 8University of living natural resources (SLU),
Uppsala, Sweden; 9University of Antwerp, Antwerp, Belgium; 10Göteborg University, Göteborg, Sweden;
11University of Zagreb, Zagreb, Croatia; 12Norwegian School of Veterinary Science (NVH), Oslo, Norway
Common questions in studies involving both the caging of fish and the sampling of wild-caught fish are
wether the measured responses are biologically significant. This is particularly difficult when evaluating
the development of effects over time and the evolution of symptoms and effects following chronic,
presumably discontinuous exposure to low concentrations of contaminants. The main objective of this
project is to identify and quantify early sublethal changes in model fish species that are predictive of
later adverse effects. The project aims to clarify the time-course of biological responses during
continuous or pulsed exposure to environmentally relevant concentrations of offshore-relevant
contaminants.
Long-term exposure experiments with zebrafish (Danio rerio) and Atlantic cod (Gadus morhua) will be
performed. In addition to providing valuable results in its own right, exposure studies with zebrafish will
be used to identify the mechanisms and endpoints to be further investigated in cod. The exposure
experiments will use a mixture of contaminants that reflect the composition of produced water. Fish will
be sampled and analysed for common biomarkers for phase 1 enzyme induction, oxidative and
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membrane-related stress, estrogenic responses, DNA integrity as well as metabolic products of PAH in
bile, histology and hatching success. Microarray and proteomics technology will also be applied. The
project will produce a larger understanding of how adverse effects can develop in fish during
contaminant exposure. Also, endpoints suitable for monitoring will be identified and an assessment of
current methods in relation to adaptation processes will be delivered.
Steinar Sanni:
Arnfinn Skadsheim: DEEP-SEA: knowledge status and methods for impact and
risk assessment
Skadsheim, A., J. F. Børseth, and S. Sanni.
RF-Akvamiljø, Mekjarvik 12, N-4070 Randaberg, Norway
For questions, contact: ask@rf.no
The Norwegian oil and gas industry is expanding northwards and into deeper waters and the nation
borders extensive deep-sea areas. Recent environmental law regulations emphasize the importance of
biological effects. In order to meet the management needs of the oil and gas industry at deep-sea
exploration and production, the development of an environmental effect and risk estimation
methodology for decision-making, impact assessment and monitoring has been initiated within the
Norwegian Deepwater Programme (NDP). As can be easily understood, there are basic aspects of
deep-sea ecotoxicology which are poorly studied, but beyond the priorities of the NDP programme to
address. This limited basic ecotoxicological knowledge in the deep-sea is addressed in our
presentation. Further, we suggest an approach to deep-sea ecotoxicology that can be useful for the oil
and gas industry’s needs. This approach, as proposed by RF-Akvamiljø, combines chemistry and
biology and adapts to limitations on the handling of live deep-sea animals so that relevant testing can be
conducted. The deep-sea is inaccessible, cold, and at high pressure but contains very species rich
benthic communities. Still, no acute toxicity tests are yet conducted on deep-sea species. Circumstantial
indices of adaptations to toxic exposures are only found in some studies at vent sites.
Due to the currently limited knowledge about the deep sea environmental risks, a precautionary
management policy will probably be required. However, in spite of a no-discharge policy, substances
may enter the environment by accidents at transport, production, pipeline leaks or larger spills. For
decision making it is therefore a demand for knowledge and monitoring methods for hazard and risk
estimation, impact assessment and follow-up of accidents and other unforeseen disturbances.
Instead of selecting just a precautionary hazard based approach due to the lack of basic
ecotoxicological knowledge about the deep sea, it should be far better to base the oil and gas
management on knowledge-based options facilitating risk assessment.
One of the important basic questions about deep sea ecotoxicology is if special adaptations in
organisms to deep-sea life may possibly also make them more vulnerable to industrial activities.
Answers to this are pertinent as to which degree we can rely on knowledge from shallow areas for deepsea environmental management.
Some of the studies we have conducted (for NDP) and descriptions of contaminant accumulation in
deep-sea food webs have raised the basic question if really high pressure (beyond 200 bars) may
significantly alter the partitioning of some hydrocarbons between oil, organisms and water and hence
possibly alter bioaccumulation and effects in biota at the great depths, such as those of the abyssal
plains.
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Odd Ketil Andersen: Proteome Expression Signatures (PES) in fish as a diagnostic
tool to evaluate the environmental impacts of offshore oil and gas exploration
Sammendrag fra prosjekt: 164413
Recent studies on fish examining the impact of produced water released offshore have revealed an
extremely complex response to oil micro droplets and oil spiked with relevant doses of poly-aromatic
hydrocarbons (PAHs) and alkylated phenols (AP). Protein expression signatures (PES) obtained from
exposed and control fish show both up and down regulation of between 50-100 proteins, with the
majority of responses observed being sex dependent, even in juvenile fish. Produced water is an
unstable mix of many components which may be unique for individual oil fields. Components in
produced water, PAHs, APs and a variety of polar organic compounds (POCs), are over represented in
comparison to concentrations found in the oil. The POCs are the least studied components of the
produced water both with respect to acute toxicity and long-term effects. In this study we will seek to
sample from cod exposure experiments financed through the PROOF, especially "Comparative oral and
water based exposures of cod to produced water components", and other programs.
Based on these findings individual proteins will be sought to be identified for further development as
biomarkers of chronic exposure to low doses of oil, synthetic mixes of PAHs, APs and POCs at
concentrations similar to those found in produced water. The proteome response in both wild and caged
cod from the North-sea will be compared against results from the laboratory exposures with the aim of
identifying and validating diagnostically important proteins. To perform the identification of diagnostically
important proteins a cooperation with national and international research groups within the field of
proteomics will be established. If possible some genomic analyses will be performed on the Cod as
cooperation with other research groups to increase the mechanistic interpretation of the proteom
responses.
Bjørn Einar Grøsvik: Effects of oil and produced water on atlantic cod; biomarker
development with proteomics based methods
Bjørn Einar Grøsvik1, Anneli Bohne Kjersem1, Sonnich Meier2, Rolf C. Sundt3, Arnfinn Skadsheim3,
Steinar Sanni3 and Anders Goksøyr1
1)
Dept. of Molecular Biology, University of Bergen, Norway
2)
Institute of Marine Research, Bergen, Norway
3)
RF-Akvamiljø, Stavanger, Norway
The substantial discharges of produced water from offshore oil and gas exploration may lead to longterm effects on pelagic fish resources like the Atlantic cod Gadus morhua L. To be better able to
address these issues cod larvae were exposed for 90 days to produced water (1%, 0.1 % and 0.01%)
and 17- estradiol (10 µg/l). This period covers newly fertilised eggs, hatching, start feeding, larvae
stage until start of feeding pellets. Maturing cod have been exposed orally to alkylated phenols and oil
and produced water compounds for 20 weeks as well as in a continuous flow for 2 weeks.
Our objectives are to utilise proteomics based techniques, i.e. 2-dimensional gel electrophoresis (2-DE)
combined with different mass spectrometry techniques and database search to study and identify
possible changes in protein expression due to such exposure. An important aim is to develop new
biomarkers to be used in future biomonitoring programmes. We also want to study possible posttranslational modifications due to such exposure. As one of the concerns of these discharges are
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endocrine disruptive effects from alkylated phenols, we will focus on changes in plasma, but also study
other organs like liver in addition to whole larvae extracts.
Preliminary results with isolated proteins from 2DE analyses of whole larvae extracts indicate differential
expression of structural proteins like actin and myosin shown by MALDI analyses after exposure to 1%
produced water.
Supported by The Norwegian Research Council, grant 164423/S40 and TOTAL E&P NORGE AS.
TEMA: Modelering
Anne Solberg:
Anders Bjørgesæter: THE DEVELOPMENT OF FIELD-BASED SPECIES
SENSITIVITY DISTRIBUTIONS (F-SSDs).
Current methods of deriving sediment predicted no effect concentrations (PNECs) or Threshold Effect
Levels (TELs) are primarily based on laboratory eco-toxicity bioassays that may not be ecologically and
environmentally relevant. In this talk I will present the methods and result from studies exploring the
possibility of utilizing field data from monitoring surveys on the marine environment on the Norwegian
Continental Shelf (the MOD-database) to construct field based species sensitivity distributions (f-SSDs)
and estimate f-PNECs. I will present how one may use quantile regression analysis to quantify and
estimate species sensitivity from scatterplots of species abundance. Because quantile regression may
estimate rates of changes across all parts of a response variable (i.e. here abundance) it is well suited
to model heterogeneous distribution like those found in benthic marine organisms occurring along an
increasing gradient of a contaminant. Results will be compared with current sediment quality guidelines
(SQG) from other parts of the world. Examples from the whole Norwegian Continental Shelf, the Ekofisk
region (a habitat) and also species (sensitive, insensitive sand opportunistic).
Henrik Rye: Utvikling av sediment modell for simulering av miljøeffekter som følge
av utslipp av borekaks og boreslam.
Bakgrunn: Oljeselskapene har gjennom et større prosjekt (ERMS = Environmental Risk Management
System) utviklet en numerisk modell for beregninger av miljøpåvirkninger som følge av utslipp av kaks
og slam på norsk sokkel. Modellen er utviklet som en del av oljeselskapenes strategi for å oppnå null
skadelige utslipp til sjø på norsk sokkel.
Metode: Modellen beregner risiko for skade på bunnsamfunn som følge av utslipp av kaks/slam på
norsk sokkel. Det beregnes 4 forskjellige typer miljøpåvirkninger: 1)Begravning (”burial”), 2)Giftighet,
3)Oksygenbrist samt 4)Forekomst av eksotisk sediment (endring i kornstørrelse). Influensområde for
påvirkning beregnes både for samlet påvirkning og for påvirkning for hver enkelt stressfaktor. Også
påvirkning i vannsøylen beregnes.
Resultater og videreføring: Modellen er så langt benyttet til å foreta noen testberegninger for aktuelle
utslipp av kaks og slam. Modellen vil nå bli benyttet til å beregne mulig påvirkninger på marin resipient
som følge av utslipp av radioaktive komponenter i produsert vann (PROOF RAIV prosjektet,
prosjektleder Dag Øistein Eriksen, FFI). Det er også søkt PROOF om støtte til et prosjekt som vil utvide
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modellen til å inkludere hvordan det aktuelle bunnsamfunn på stedet kan påvirke modellresultatene
(PROOF PEIOFF - FAME, prosjektet, prosjektleder Frode Olsgaard, NIVA).
Frode Olsgard: Parameterisation of the Environmental Impacts on Bottom Fauna
of Water-based Drilling Fluids and Cuttings – Field and Mesocosm Experiments
(PEIOFF-FAME)
Authors: Dr. Frode Olsgard, NIVA; Dr. Henrik Rye, SINTEF og cand. scient Hilde Cecilie Trannum,
Akvaplan-NIVA
In Norway, monitoring surveys aimed at studying possible effects of disturbance on the sedimentdwelling fauna from offshore oil and gas exploration activities and disposal of drill cuttings have been a
standard since the early 1970s. Drill cuttings discharges have caused significant ecological changes in
the benthic fauna (eg. reduction in number of species, abundance and diversity) adjacent to many
platforms in the North Sea. Today water based drilling muds (WBM) are the main drilling mud used and
adverse effects on the bottom fauna are considered to be the burying effect on sessile benthic species,
changes in the surface sediment grainsize, development of anoxia in the surface sediments and
possible toxic effects on the benthic fauna from drilling mud compounds. Over the last years the oil
companies has initiated the development of an environmental risk model (ERMS) to predict and
evaluate effects of drill cutting discharges. The model have till now mainly been based on
physical/chemical process, without incorporating the effects the bottom fauna has on the various
parameters used in the model. It is important to acknowledge that the benthic fauna is both affected by
and will affect the WBM drill cutting discharges. In this project data will, through controlled mesocosmand field studies and literature studies provide quantitative information needed for development of a
Faunal Module in ERMS and quantitatively describe the adverse effects of WBM on bottom fauna and
the fauna recolonisation processes on WBM cutting disposals.
Renee Katrin Bechmann: Effekter av boreslam på torsk og skjell
Bechmann, R.K., Baussant, T., Taban, I.C., Øysæd, K.B., Pampanin, D., Lowe, D., Sundt, R.C.,
Westerlund, S.
De to viktigste målsetningene er å finne ut om eksponering for suspenderte boreslampartikler gir
negative effekter på dyr som lever i de fri vannmassene (fisk og plankton) og skjell som filtrerer vannet
for å skaffe seg mat, og om metaller akkumuleres i skjell og torsk som følge av eksponeringen.
Mengden barium i torsk og skjell økte med økende eksponering av brukt boreslam med barytt som
vektstoff. Mengden av enkelte andre metaller økte også, men vi kan ikke utelukke at disse metallene var
bundet i baryttpartikler i vevet. Resultatene vil bli diskutert i forhold til akkumulering av metall og effekter
i torsk og skjell eksponert for en blanding av metaller løst i vann. Tre ukers eksponering for ilmenitt (kun
torsk), barytt og brukt boreslam med barytt ga histologiske skader i skjell og torsk. Metallblandingen ga
også histologiske skader, og vi har tidligere sett at både boreslam og metall ga redusert
filtreringseffektivitet og redusert vekst for skjell eksponert i 6-10 uker. Signifikant lavere
filtreringseffektivitet ble observert for kamskjell eksponert for 0.5, 2, og 20 mg/L brukt boreslam. Vi har
gjort to forsøk med blåskjellarver eksponert for boreslam. Det første forsøket viste at andelen
deformerte larver økte når de ble eksponert for brukt boreslam. Årets forsøk viste at boreslam førte til at
larvene spiste mindre effektivt enn larver i rent vann. Overlevelse og vekst av torskelarver ble ikke
redusert etter to ukers eksponering for 6 mg/L brukt boreslam med barytt. Det vil bli gitt en oversikt over
resultater fra alle forsøkene i prosjektet.
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JoLynn Carroll: Environmental effects of offshore oil activities: Experimental tests
of petroleum-associated components on benthos at community, individual, and
cellular levels
Gro Harlaug Olsen1, JoLynn Carroll1, Michael Carroll1, Ragni Olssøn1, Lionel Camus1, Will Ambrose1,
Ketil Hylland2, Morten Schaaning2, Frode Olsgard2.
1Akvaplan-niva
2Norwegian
AS, Polar Environmental Center, 9296 Tromsø
Institute for Water Research, 0411 Oslo
To responsibly apply monitoring techniques developed in the North Sea to other regions, we must know
the effects on benthic communities from oil-related activities and the ecosystem processes controlling
the transfer of petroleum-associated components between pelagic and benthic ecosystems. For this
purpose, a series of controlled experiments are being performed to identify benthic community
responses to routine discharges of drill cuttings and to evaluate effects on benthic organisms (organismand cellular-level) from acute spills of petroleum-associated chemicals. Experimental results are being
combined with an examination of real-world monitoring data sets collected under Norway’s Petroleum
Regional Monitoring Programme in order to identify the geographic scope of responses to petroleum
industrial activities. At the completion of the project, the results will be used to evaluate whether
indicators, techniques, and threshold levels used for environmental assessment of the impact of
petroleum activities at lower latitudes are appropriate for the arctic, and propose improvements to the
interpretation of benthic monitoring data from diverse environmental regions. These efforts will result in
unique data for use in validating and testing the benthic module of the Environmental Risk Management
System (ERMS).
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