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Parallel monitoring of mixed-functional oxygenase (MFO) induction and DNA damage in marine organisms
Abstracts
355
Imposex phenomena did occur in some female whelks, which showed tiny or somewhat bigger penis homologues. At some Northern most stations no imposex phenomena occurred. At the other Northern stations the phenomena were smaller and the incidence was lower than in the central and southern North Sea. The highest rates of imposex in the central and southern North Sea coincided with the vicinity of shipping lanes. The imposex rates were higher in the more densely shipped routes than in the less busy north-south deep water route. The station “Brucey’s Garden” a supposed reference area in the northern North Sea, that showed somewhat elevated incidence of imposex, appeared to be situated in a relatively more intensely used shipping area than the Dogger Bank, where only one minor imposex phenomenon was found. The levels of tributyltin (TBT) and its metabolites, dibutyltin (DBT), and monobutyltin (MBT), were measured in whelks from the first leg of the cruise. TBT was not detectable, but DBT and MBT were, albeit at low levels. At the only station where all females showed imposex, DBT and MBT levels were twice as high as in the other stations where DBT and MBT were detectable. Metabolization of TBT in Buccinum undatum may differ from that in Nucella lapillus, the snail in which imposex was clearly related to elevated levels of TBT in animals and environment. Laboratory experiments to try and link the whelks imposex phenomena to the presence of TBT and metabolites from anti-fouling paints are under way.
Parallel Monitoring of Mixed-Functional Oxygenase (MFO) Induction and DNA Damage in Marine Organisms. A. HERBERT, D. WARNECKE, H. KRUMBECK & P.-D. HANSEN. Berlin University of Technology, Institute of Ecology, Dept. of Aquatic
Ecotoxicology,
Goerzallee
253, D-W-1000
Berlin 37, Germany.
Induction of MFO activities in marine fish is now widely used as a biomarker indicating exposure to organic pollutants such as polycyclic aromatic hydrocarbons or chlorinated aromatic hydrocarbons. Although MFOs are part of a detoxifying system, they generate reactive intermediate metabolites; which may damage the primary biological information matrix DNA. Since studies of MFO induction in fish may suffer from interferences of parameters like, e.g. sex and season, it appears useful to supplement MFO monitoring with parallel monitoring of DNA damage. Rather than being simply an indicating parameter for genotoxic exposure, the genotoxic event itself is detected and quantified. To provide an instrument for large-scale monitoring of DNA damage, the authors have adapted a method from cell culture research - an alkaline DNA unwinding assay - to the study of marine organisms, which fits the following requirements: (a) wide spectrum of sensitivites providing a sum parameter for different types of DNA lesions; (b) high cost efficiency; (c) easy use; (d) high sample throughput; and (e) portability for the use in field studies, e.g. on research vessels. The non-radioactive method has been successfully used since 1985 in laboratory and field studies, both in marine invertebrates and fish. Results indicate, that fish might be more sensitive to some types of pollutants, possibly due to different capacities of the MFO system. In a case study, DNA damage in liver of fish from
356
Abstracts
a wild-life population coincided with an induction of mixed-functional oxygenase (MFO) activity, when compared to fish held in laboratory tanks with filtered sea-water.
Ecotoxicologic Studies Investigating the Decline of the Striped Bass (Morone suxafibs) in the Sacramento and San Joaquin Rivers and Delta-Bay Estuary. D. E. HINTON, H. R. BAILEY, W. 0. BENNETT & D. G. OSTRACH. Department of Medicine, School of Veterinary .fornia 9.5616, USA.
Medicine,
University
of California,
Davis, Cali-
Striped bass were first bought to California from the eastern US in 1879. The population underwent steady growth until the last 20 years. California State Game and Fish Commission biologists have constructed an annual striped bass larval recuitment index. The recruitment index has declined annually and all workers are in agreement that the population is in a decline. Factors that could lead to a lack of recuitment include the effects of large volumes of water being diverted from the river system, toxic effects of rice field chemicals, the entrainment and entrapment of pelagic larvae in pumps and diversion canals, and an inadequate food source. Three years of data on field collections have indicated that food organisms are present in the guts of larval fish and companion morphometric and histopathologic investigations have indicated that lesions, produced in the laboratory by starvation, are not seen in field fish. Although the quality of the food has not been determined at this time, the finding of lesions suggestive of toxic chemicals in brain and liver has led us to conduct investigations on the toxicity of water from rice fields. Grab samples from the Colusa Basin Drain, a large single point source of rice field waste-water entry into the Sacramento River have been investigated for their toxicity in striped bass and invertebrates. The possible role of these toxicants and their effects on striped bass recruitment and further growth will be presented.
Effects of PBDEs, PCNs and PCBs on Reproduction, Liver Morphology and Cytochrome P450 Activity in the Three-Spined Sticklehack, Gusterosteus myleatus. G. HOLM,“,b L. NORRGREN,“,’ T. ANDERSSONd & A. THUREN.” “Swedish Environmental Research Institute, Box 21060, S-100 31 Stockholm, Sweden; bDepartment of Zoology, University of Stockholm, S-106 91 Stockholm, Sweden; ‘Department of Pathology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, S-750 07 Uppsala, Sweden; dDepartment of Zoophysiology, University of Goteborg, Medicinaregatan 18, S-413 90 Gd’teborg, Sweden.
Female three-spined sticklebacks were fed with freeze-dried chironomids contaminated with low or high doses of polybrominated diphenyl ethers (PBDEs) (Bromkal 70-5DE), polychlorinated naphthalenes (PCNs) (Halowax 1014) and polychlorinated biphenyls (PCBs) (Clophen A50). After 3.5 months of exposure, reproduction studies were performed. No significant difference in number of eggs laid was found between the control and exposed groups. Spawning success in the control group was 80% whereas it was 20% in the groups that received high
355
Imposex phenomena did occur in some female whelks, which showed tiny or somewhat bigger penis homologues. At some Northern most stations no imposex phenomena occurred. At the other Northern stations the phenomena were smaller and the incidence was lower than in the central and southern North Sea. The highest rates of imposex in the central and southern North Sea coincided with the vicinity of shipping lanes. The imposex rates were higher in the more densely shipped routes than in the less busy north-south deep water route. The station “Brucey’s Garden” a supposed reference area in the northern North Sea, that showed somewhat elevated incidence of imposex, appeared to be situated in a relatively more intensely used shipping area than the Dogger Bank, where only one minor imposex phenomenon was found. The levels of tributyltin (TBT) and its metabolites, dibutyltin (DBT), and monobutyltin (MBT), were measured in whelks from the first leg of the cruise. TBT was not detectable, but DBT and MBT were, albeit at low levels. At the only station where all females showed imposex, DBT and MBT levels were twice as high as in the other stations where DBT and MBT were detectable. Metabolization of TBT in Buccinum undatum may differ from that in Nucella lapillus, the snail in which imposex was clearly related to elevated levels of TBT in animals and environment. Laboratory experiments to try and link the whelks imposex phenomena to the presence of TBT and metabolites from anti-fouling paints are under way.
Parallel Monitoring of Mixed-Functional Oxygenase (MFO) Induction and DNA Damage in Marine Organisms. A. HERBERT, D. WARNECKE, H. KRUMBECK & P.-D. HANSEN. Berlin University of Technology, Institute of Ecology, Dept. of Aquatic
Ecotoxicology,
Goerzallee
253, D-W-1000
Berlin 37, Germany.
Induction of MFO activities in marine fish is now widely used as a biomarker indicating exposure to organic pollutants such as polycyclic aromatic hydrocarbons or chlorinated aromatic hydrocarbons. Although MFOs are part of a detoxifying system, they generate reactive intermediate metabolites; which may damage the primary biological information matrix DNA. Since studies of MFO induction in fish may suffer from interferences of parameters like, e.g. sex and season, it appears useful to supplement MFO monitoring with parallel monitoring of DNA damage. Rather than being simply an indicating parameter for genotoxic exposure, the genotoxic event itself is detected and quantified. To provide an instrument for large-scale monitoring of DNA damage, the authors have adapted a method from cell culture research - an alkaline DNA unwinding assay - to the study of marine organisms, which fits the following requirements: (a) wide spectrum of sensitivites providing a sum parameter for different types of DNA lesions; (b) high cost efficiency; (c) easy use; (d) high sample throughput; and (e) portability for the use in field studies, e.g. on research vessels. The non-radioactive method has been successfully used since 1985 in laboratory and field studies, both in marine invertebrates and fish. Results indicate, that fish might be more sensitive to some types of pollutants, possibly due to different capacities of the MFO system. In a case study, DNA damage in liver of fish from
356
Abstracts
a wild-life population coincided with an induction of mixed-functional oxygenase (MFO) activity, when compared to fish held in laboratory tanks with filtered sea-water.
Ecotoxicologic Studies Investigating the Decline of the Striped Bass (Morone suxafibs) in the Sacramento and San Joaquin Rivers and Delta-Bay Estuary. D. E. HINTON, H. R. BAILEY, W. 0. BENNETT & D. G. OSTRACH. Department of Medicine, School of Veterinary .fornia 9.5616, USA.
Medicine,
University
of California,
Davis, Cali-
Striped bass were first bought to California from the eastern US in 1879. The population underwent steady growth until the last 20 years. California State Game and Fish Commission biologists have constructed an annual striped bass larval recuitment index. The recruitment index has declined annually and all workers are in agreement that the population is in a decline. Factors that could lead to a lack of recuitment include the effects of large volumes of water being diverted from the river system, toxic effects of rice field chemicals, the entrainment and entrapment of pelagic larvae in pumps and diversion canals, and an inadequate food source. Three years of data on field collections have indicated that food organisms are present in the guts of larval fish and companion morphometric and histopathologic investigations have indicated that lesions, produced in the laboratory by starvation, are not seen in field fish. Although the quality of the food has not been determined at this time, the finding of lesions suggestive of toxic chemicals in brain and liver has led us to conduct investigations on the toxicity of water from rice fields. Grab samples from the Colusa Basin Drain, a large single point source of rice field waste-water entry into the Sacramento River have been investigated for their toxicity in striped bass and invertebrates. The possible role of these toxicants and their effects on striped bass recruitment and further growth will be presented.
Effects of PBDEs, PCNs and PCBs on Reproduction, Liver Morphology and Cytochrome P450 Activity in the Three-Spined Sticklehack, Gusterosteus myleatus. G. HOLM,“,b L. NORRGREN,“,’ T. ANDERSSONd & A. THUREN.” “Swedish Environmental Research Institute, Box 21060, S-100 31 Stockholm, Sweden; bDepartment of Zoology, University of Stockholm, S-106 91 Stockholm, Sweden; ‘Department of Pathology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, S-750 07 Uppsala, Sweden; dDepartment of Zoophysiology, University of Goteborg, Medicinaregatan 18, S-413 90 Gd’teborg, Sweden.
Female three-spined sticklebacks were fed with freeze-dried chironomids contaminated with low or high doses of polybrominated diphenyl ethers (PBDEs) (Bromkal 70-5DE), polychlorinated naphthalenes (PCNs) (Halowax 1014) and polychlorinated biphenyls (PCBs) (Clophen A50). After 3.5 months of exposure, reproduction studies were performed. No significant difference in number of eggs laid was found between the control and exposed groups. Spawning success in the control group was 80% whereas it was 20% in the groups that received high