Gunilla Ericson

Biochemical indicators of pollution exposure in shorthorn sculpin (Myoxocephalus scorpius), caught in four harbours on the southwest coast of Iceland

Shorthorn sculpins (Myoxocephalus scorpius) were caught in four Icelandic harbours, differing in size, use and traffic. Biochemical responses in liver were measured and chemicals analysed in bile. Eyrarbakki harbour, which has not been in use for many years was chosen as a control site. Njar partial differentialvík harbour is a small fishing harbour and a marina, Sandger partial differentiali harbour is a large fishing harbour, and Reykjavík harbour is a large fishing harbour and an international transport harbour. Higher levels of DNA-adducts and cytochrome P4501A (CYP1A) in the fish from the harbours in Sandger partial differentiali, Njar partial differentialvík and Reykjavík, compared to Eyrarbakki harbour, indicate PAH exposure. This was confirmed by PAH analysis in bile. The higher activities of the antioxidant enzymes catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) in fish caught in Sandger partial differentiali, than in fish caught in the other harbours, indicate exposure of sculpin to prooxidative compounds in Sandger partial differentiali harbour. Shorthorn sculpin seems to be a convenient species for monitoring pollution in northern coastal areas.

Effects of two polybrominated diphenyl ethers on rainbow trout (Oncorhynchus mykiss) exposed via food

Abstract Two polybrominated diphenyl ethers, 2, 4, 2′, 4′-tetrabromodiphenyl ether and 2, 4, 5, 2′, 4′-pentabromodiphenyl ether were added to the food given to two different groups of rainbow trout. The rainbow trout were fed for a period of 22 days. On days 6 and 22, a number of physiological and biochemical variables were investigated including condition factor, liver somatic index, spleen somatic index, hematocrit, differential count of leucocytes, haemoglobin and glucose in whole blood as well as the liver enzyme activities, glutathione reductase, catalase and ethoxyresorufin-O-deethylase. The results show that both these substances were relatively non-acutely toxic and several variables were unaffected. However, the blood variables hematocrit and blood glucose showed a slight influence from 2, 4, 5, 2′, 4′-pentabromodiphenyl ether after six days of exposure. Also the glutathione reductase activity in the liver decreased after 22 days of exposure. The 2, 4, 2′, 4′-tetrabromodiphenyl ether showed no influence on the investigated variables, with one striking exception. This structure caused a decrease of about 75% in ethoxyresorufin-O-deethylase activity in the liver after six days, and this effect was also observed after 22 days.

DNA adduct formation and persistence in liver and extrahepatic tissues of northern pike (Esox lucius) following oral exposure to benzo[a]pyrene, benzo[k]fluoranthene and 7H-dibenzo[c, g]carbazole

The formation and persistence of DNA adducts in liver, intestinal mucosa, gills and brain of juvenile northern pike (Esox lucius) following oral exposure to benzo[a]pyrene (BaP), benzo[k]fluoranthene (BkF) and 7H-dibenzo[c,g]carbazol (DBC) were analysed by 32P-postlabelling. The dosage was 25 micromol/kg body weight of each substance, administered on 5 occasions with an interval of 12-14 days. Sampling was carried out 9 days after the second treatment, and 9, 16, 33 and 78 days after the fifth treatment. Pikes were also fed with the substances singly for comparison of adduct patterns. A complex pattern of adducts was detected in all examined tissues from fish treated with the mixture. Total adduct levels were highest in intestine (347+/-17.4 nmol adducts/mol nucleotides, mean+/-SE), followed by liver (110+/-9.3), gills (69+/-6) and brain (14+/-4.2). In pike treated with BaP alone, one major adduct was detected in all examined tissues. This BaP-adduct made up approximately 50% of the total amount of adducts in the brain. Corresponding values in liver, intestine and gills were 23, 31 and 34%, respectively. One relatively weak BkF-adduct and at least 10 different DBC-adducts were detected in all analysed tissues. Total adduct level in the intestine declined to 29.4% of the maximum value 78 days after the last exposure, while there was no significant decline in adduct levels in liver, gills or brain. The results suggest that intestine is more susceptible to adduct formation than liver after oral exposure, and that adduct levels in the intestine represent ongoing or relatively recent exposure. DNA adducts in the other investigated tissues were much more persistent and may therefore accumulate during long-term exposure.

Tissue differences, dose-response relationship and persistence of DNA adducts in blue mussels (Mytilus edulis L.) exposed to benzo[a]pyrene

Baltic Sea blue mussels (Mytilus edulis) were experimentally exposed to the genotoxic model substance benzo[a]pyrene (B[a]P) to study DNA adduct formation. The specific aims were (a). to examine where in the mussels the DNA adducts were formed, in gills or digestive gland; (b). to study the dose-response relationship between B[a]P exposure and DNA adduct formation; and (c). to examine the persistence of the formed adducts. A Scope for growth (SFG) study was also run to compare physiological responses of the mussels with the degree of DNA adduct formation. In an initial dose-response experiment, the mussels were exposed to 0, 5, 50, and 100 microg/l of tritium labelled B[a]P under semi-static conditions for 4 days, and thereafter the bioaccumulation of B[a]P and DNA adduct formation in different tissues was determined using liquid scintillation counting and 32P-postlabelling analysis, respectively. In a following exposure-depuration experiment, mussels were exposed to 17 microg/l of radiolabelled B[a]P under semi-static conditions for 6 days. B[a]P accumulation and DNA adduct formation were determined during the exposure, and B[a]P elimination and persistence of DNA adducts were studied during 28 days of depuration in uncontaminated water. The results revealed large tissue differences in DNA adduct formation. DNA adduct levels were not elevated in the digestive gland of the mussels at any exposure concentration (0-100 microg/l), even though the highest B[a]P tissue concentrations were found in the digestive gland (1.0+/-0.1 mg B[a]P/g tissue dry wt at 100 microg/l, mean+/-SE, n=12). DNA adducts were on the other hand formed in the gills, with the highest levels found in mussels exposed to 50 and 100 microg B[a]P/l, and a dose dependent increase in adduct levels (from 1.6 to 5.9 nmol adducts/mol nucleotides) from 0 to 50 microg B[a]P/l. In gills, DNA adduct levels increased with time during the 6-day exposure period in the exposure-depuration experiment, and then persisted for at least 2 weeks after exposure cessation while B[a]P tissue levels exhibited a rapid decrease (half-life of 8 days). No significant differences were observed in SFG between the control and exposed groups. Since DNA adducts exhibited a relatively high persistence in gills compared to B[a]P tissue concentrations, they seem to be a more integrated measure of genotoxic exposure than only chemical analysis of the contaminant bioaccumulation. The results also suggest that if using analysis of DNA adducts in M. edulis for monitoring purposes, analysis of gills in addition to the more commonly used digestive gland should be taken into consideration.

Accumulation and effects of aluminum smelter-generated polycyclic aromatic hydrocarbons on soft-bottom invertebrates and fish

An integrated study involving measurements of polycyclic aromatic hydrocarbon (PAH) levels in bottom sediments, assessments of resident soft-bottom communities, the accumulation of PAHs in soft-bottom invertebrates, and biomarker responses in invertebrates and fish was conducted to assess the impact of an aluminum reduction plant in a Norwegian fjord. The fjord sediments were heavily contaminated by PAHs in the inner reaches near the aluminum smelter, where concentrations were well above levels elsewhere reported to induce biological effects. Nevertheless, the PAH contamination in the fjord did not seem to have severe effects on the benthic biota. This conclusion can be drawn from the soft-bottom communities as well as from biomarker analyses. Presumably, contaminant speciation is important for explaining the restricted biological effects. The results support the assumption that PAHs associated with soot-like structures have limited bioavailability. They also point to the need to link various single-species approaches to measurements of effects on higher levels of organization and with an understanding of the speciation of the chemical contaminant.

DNA adduct formation in northern pike (Esox lucius) exposed to a mixture of benzo[a]pyrene, benzo[k]fluoranthene and 7H-dibenzo[c, g]carbazole: time-course and dose-response studies

The time-course and dose dependent formation of DNA adducts in juvenile northern pike (Esox lucius) following a single exposure to a mixture of benzo[a]pyrene (BaP), benzo[k]fluoranthene (BkF) and 7H-dibenzo[c,g]carbazole (DBC) were investigated by use of the (32)P-postlabelling assay. A complex adduct pattern was detected in liver and intestine of exposed fish. For the time-course studies fish were exposed either by oral administration or by intraperitoneal (i.p.) injection. Following a single i.p. injection of the mixture (40micromole/kg body weight of each substance) significantly elevated DNA adduct levels were detected in the liver after 1 day. Adduct levels were higher in liver than in intestine, in which significant elevation were detected from day 3 to 12. Following exposure via food (80micromole/kg body weight of each substance), adduct levels were detected in both liver and intestine 1 day after exposure, and continued to increase until day 3 in liver and day 6 in intestine. Calculation of a binding index, which compensates for differences in dosage, resulted in much higher adduct formation (five times in liver and 22 times in intestine) following oral exposure. Pikes receiving single oral doses of 12.5, 50, 100 or 200micromole/kg body weight of each substance exhibited significantly higher adduct levels in both liver and intestine compared to controls. Hepatic adduct levels were also higher in fish given 100 and 200micromole/kg compared to 12.5micromole/kg. Results from this study show that DNA adducts are rapidly formed in juvenile northern pike following both i.p. injection and feeding of a mixture of BaP, BkF and DBC. A maximum level was reached within a few days, which then persisted at approximately the same level for at least 9-12 days. The results also shows that higher levels of adducts were obtained following oral administration compared to i.p. injection, particularly in the intestine.

DNA adducts in perch (Perca fluviatilis) from a creosote contaminated site in the River Ångermanälven, Sweden

Levels of hepatic DNA adducts, analysed by 32P-postlabelling, histopathological lesions and organosomatic indices, were measured in perch (Perca fluviatilis). The fish were caught at five sites in the river Angermanalven, Sweden: one site known to be contaminated with creosote, sites downstream from the contaminated site and a reference site upstream. Perch were also caught at a long-distance reference site. The level of DNA adducts in fish from the creosote-contaminated site was 6.8±4.1 nmol mol−1 nucleotides compared to 0.21±0.21 nmol mol−1 nucleotides in fish from the long-distance reference site. The adduct level was also significantly increased compared to adduct levels in fish from the local reference site. Hepatocellular degeneration and macrophage aggregates were observed but did not correlate with a specific site in the river. No effects on organosomatic indices that correlated with distance from the contaminated site were observed. In the laboratory, perch were exposed to an organic solvent extract prepared from sediment collected at the creosote-contaminated site or to benzo(a)pyrene (BaP) by oral administration. Perch treated with the extract had adduct patterns very similar to those observed in perch from the contaminated field site. Two diagonal zones including 10–12 adduct spots were observed on the autoradiograms. One adduct was tentatively identified by co-chromatography with a 32P-labelled standard of 7R,8S,9S-trihydroxy, 10R-(N2-deoxyguanosyl 3′-phosphate)-7,8,9,10-tetrahydrobenzo(a)pyrene (BaPDE-dG-3′p) in two different solvent systems. Autoradiograms derived from the BaP-exposed perch showed one major adduct and several less intense spots. The major BaP adduct had the same chromatographic properties as the standard BaPDE-dG-3′p adduct. This study shows that 32P-postlabelling analysis of DNA adducts in perch can be used as a sensitive indicator of exposure to genotoxic polycyclic aromatic hydrocarbons, and that comparison of DNA adduct patterns of fish exposed in the field and laboratory can be used to determine the responsible source of pollutants.

Further studies of the effects of exhaust from two-stroke outboard motors on fish

Abstract Rainbow trout were exposed to an n -hexane extract of engine exhaust condensate by intra peritoneal (i.p.) injection or by feeding with contaminated home-made cod chips. Exposure levels were calculated and chosen to reflect those conditions possibly experienced by fish in their natural habitats. Disruption of biological functions has been observed at different levels of biological organization including cellular and subcellular processes (DNA-adduct levels and enzyme activity) and physiological functions (carbohydrate metabolism). Female and male fish may be affected to a varying degree by these types of pollutants.

Investigation of the Biological Effects of 2-Cycle Outboard Engines' Exhaust on Fish

Abstract A 2-cycle outboard engine was run in a tank to simulate boatload conditions. After appropriate dilution, exhaust water was led into another tank where fish were exposed. In addition, exposure from extract obtained by acetone/hexane extracted water containing exhaust was used in intraperitoneal injection of juvenile fish as well as in embryo injection experiments. The different areas of investigation were enzymatic disturbances, genotoxicological effects and reproduction disturbances. The investigation so far indicates that the biological effects of the exhaust from 2-cycle outboard engines is a serious threat to the environment. Toxicological effects have been measured in the liver, kidney and blood.

Comparative32P-postlabeling analysis of DNA Adducts in Perch (Perca fluviatilis) from unpolluted and polluted areas in Swedish coastal waters

Abstract Perch (Perca fluviatilis), were sampled from unpolluted and polluted areas in Swedish coastal waters. The level of aromatic/hydrophobic DNA adducts in liver tissue was analyzed using the nucleas P1 version of the32P-post-labeling assay. The level of total adducts measured in the individual fish from polluted areas was between 6 and 22 nmol of adducts/mol of nucleotides, and in the fish from the reference area between ⩽ 0.2 and 0.6 nmol of adducts/mol of nucleotides.