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Kinetics and magnitude of metallothionein induction by Cd, Cu, Hg and Zn in European flounder — calibration for environmental monitoring
Abstracts / Marine Environmental Research 50 (2000) 431±441
433
haemocytes eciently take up BFLV to their lysosomes, while uptake is decreased in haemocytes of exposed mussels. EthD-1 is taken up generally with time and the ¯uorescence will increase upon binding to nucleic acids; however, damaged cells take up EthD-1 to a greater extent. Thereby, the ratio of the ¯uorescent probes enhances the signal. The eect was veri®ed by ¯uorescence microscope studies. The results with pyrene demonstrated signi®cant reduction in ratio of ¯uorescence after treatment with 0.4, 1, and 2.5 mg/l for 1 week, as did 2 mg/l Cu and Zn. The results have been compared with the NRR assay, demonstrating similar sensitivity. A comparison of the methods has also been performed with mussels collected from a metal-polluted ®eld site. The measured ¯uorescence ratio may be an alternative to the NRR assay, providing automated reading. PII: S0141-1136(00)00205-1
Kinetics and magnitude of metallothionein induction by Cd, Cu, Hg and Zn in European ¯ounder Ð calibration for environmental monitoring S. George a, J. Wright a, E. Carpene b, M. Kindt b a Institute of Aquaculture, University of Stirling, Stirling FKL9 4LA, UK Department of Biochemistry FM Vet, University of Bologna, Ozzano Emilia, Bologna, Italy
b
Abstract The inductive response of metallothionein (MT) synthesis to heavy-metal exposure in many Phyla is well documented; however, before this response can be used and interpreted as an environmental biomarker of exposure, the kinetics and dose responses must be known. We have carried out a series of experiments where we have studied these parameters of MT mRNA and MT protein induction in dierent tissues (liver, kidney, gills) of European ¯ounder after intraperitoneal (ip) injection of Cd (1±320 mg/kg), Cu (25±200 mg/kg), Hg (235±100 mg/kg) and Zn (1±20 mg/kg). Basal hepatic levels of 320 mg MT/g wet tissue for the wild caught ¯ounder used were ®ve times higher than in ®sh from a pristine area and 10±20 times higher than in plaice or turbot obtained from pristine areas. Transient increases in renal MT mRNA levels were only observed 4±10 days post injection (p.i.) of Cu or Hg and there were no signi®cant increases in MT protein levels in any tissue. Doses of >5 mg/kg Zn induced MT mRNA levels three- to four-fold in all tissues 2±4 days p.i. but only produced a transient elevation in MT levels at 4 days p.i. There was no measurable response to low doses of Cd, doses of >80 mg/kg Cd had no signi®cant eect on hepatic MT levels but caused a signi®cant elevation in gill and kidney levels 10 and 21 days p.i. MT mRNA levels in gill and kidney were
434
Abstracts / Marine Environmental Research 50 (2000) 431±441
induced four- to ®ve-fold 24 h p.i. of Cd (and remained elevated for >10 days) whilst a response was only observed in liver (two-fold) 4 and 10 days p.i. These laboratory results, obtained with a wild population exhibiting moderately elevated basal MT levels indicate that the response to metal exposure is too small and too highly dependent upon the metal and exposure time to provide a robust biomarker of contaminant eect. This supports our previous observation of analyses of ¯ounders from the southern North Sea and Dutch estuaries and would indicate that caging studies with `ultra-clean' animals would be required for any monitoring studies. PII: S0141-1136(00)00206-3
Proliferative and neoplastic conditions of ¯ounder (Platichthys ¯esus) in the northern Baltic Sea in relation to DNA-adducts G. Bylund a, C. MalmstroÈm a, T. Wiklund a, M.S. Myers b a Institute of Parasitology, AÊbo Akademim University, AÊbo, Finland Environmental Conservation Division, Northwest Fisheries Center, National Marine Fisheries Service/NOAA, Seattle, WA 98112, USA
b
Abstract A disease survey o the Finnish sea coast of the Baltic Sea included 6644 specimens of ¯ounder (Platichthys ¯esus). The aim of the survey was to explore to what extent the quality of the coastal environment in the area is re¯ected in ®sh health parameters. As degenerative and proliferative conditions of ®sh liver have increasingly been recognized to be most closely correlated with chemical contaminants special attention was given to this organ. Some neoplastic conditions were recorded in other organs/tissues in the ¯ounders and are reported too. On an average 3.0% of all ¯ounders examined (>20 cm) were aected by preneoplastic/ neoplastic liver lesions. The prevalence is signi®cantly increasing with increasing size of ®sh. Of specimens >30 cm 10.1% were aected and, at one site the disease prevalence in female ¯ounder >30 cm was 31.5%, which must be considered strikingly high. The site not immediately in¯uenced by human activities but in closest contact with the main Baltic Sea, had the highest lesion prevalences. The levels of hydrophobic DNA-adducts in the liver of ¯ounder in the area are low which indicates that, for example, polycyclic aromatic hydrocarbon compounds are not responsible for the high prevalences of proliferative lesions in ¯ounder in this area. PII: S0141-1136(00)00207-5
433
haemocytes eciently take up BFLV to their lysosomes, while uptake is decreased in haemocytes of exposed mussels. EthD-1 is taken up generally with time and the ¯uorescence will increase upon binding to nucleic acids; however, damaged cells take up EthD-1 to a greater extent. Thereby, the ratio of the ¯uorescent probes enhances the signal. The eect was veri®ed by ¯uorescence microscope studies. The results with pyrene demonstrated signi®cant reduction in ratio of ¯uorescence after treatment with 0.4, 1, and 2.5 mg/l for 1 week, as did 2 mg/l Cu and Zn. The results have been compared with the NRR assay, demonstrating similar sensitivity. A comparison of the methods has also been performed with mussels collected from a metal-polluted ®eld site. The measured ¯uorescence ratio may be an alternative to the NRR assay, providing automated reading. PII: S0141-1136(00)00205-1
Kinetics and magnitude of metallothionein induction by Cd, Cu, Hg and Zn in European ¯ounder Ð calibration for environmental monitoring S. George a, J. Wright a, E. Carpene b, M. Kindt b a Institute of Aquaculture, University of Stirling, Stirling FKL9 4LA, UK Department of Biochemistry FM Vet, University of Bologna, Ozzano Emilia, Bologna, Italy
b
Abstract The inductive response of metallothionein (MT) synthesis to heavy-metal exposure in many Phyla is well documented; however, before this response can be used and interpreted as an environmental biomarker of exposure, the kinetics and dose responses must be known. We have carried out a series of experiments where we have studied these parameters of MT mRNA and MT protein induction in dierent tissues (liver, kidney, gills) of European ¯ounder after intraperitoneal (ip) injection of Cd (1±320 mg/kg), Cu (25±200 mg/kg), Hg (235±100 mg/kg) and Zn (1±20 mg/kg). Basal hepatic levels of 320 mg MT/g wet tissue for the wild caught ¯ounder used were ®ve times higher than in ®sh from a pristine area and 10±20 times higher than in plaice or turbot obtained from pristine areas. Transient increases in renal MT mRNA levels were only observed 4±10 days post injection (p.i.) of Cu or Hg and there were no signi®cant increases in MT protein levels in any tissue. Doses of >5 mg/kg Zn induced MT mRNA levels three- to four-fold in all tissues 2±4 days p.i. but only produced a transient elevation in MT levels at 4 days p.i. There was no measurable response to low doses of Cd, doses of >80 mg/kg Cd had no signi®cant eect on hepatic MT levels but caused a signi®cant elevation in gill and kidney levels 10 and 21 days p.i. MT mRNA levels in gill and kidney were
434
Abstracts / Marine Environmental Research 50 (2000) 431±441
induced four- to ®ve-fold 24 h p.i. of Cd (and remained elevated for >10 days) whilst a response was only observed in liver (two-fold) 4 and 10 days p.i. These laboratory results, obtained with a wild population exhibiting moderately elevated basal MT levels indicate that the response to metal exposure is too small and too highly dependent upon the metal and exposure time to provide a robust biomarker of contaminant eect. This supports our previous observation of analyses of ¯ounders from the southern North Sea and Dutch estuaries and would indicate that caging studies with `ultra-clean' animals would be required for any monitoring studies. PII: S0141-1136(00)00206-3
Proliferative and neoplastic conditions of ¯ounder (Platichthys ¯esus) in the northern Baltic Sea in relation to DNA-adducts G. Bylund a, C. MalmstroÈm a, T. Wiklund a, M.S. Myers b a Institute of Parasitology, AÊbo Akademim University, AÊbo, Finland Environmental Conservation Division, Northwest Fisheries Center, National Marine Fisheries Service/NOAA, Seattle, WA 98112, USA
b
Abstract A disease survey o the Finnish sea coast of the Baltic Sea included 6644 specimens of ¯ounder (Platichthys ¯esus). The aim of the survey was to explore to what extent the quality of the coastal environment in the area is re¯ected in ®sh health parameters. As degenerative and proliferative conditions of ®sh liver have increasingly been recognized to be most closely correlated with chemical contaminants special attention was given to this organ. Some neoplastic conditions were recorded in other organs/tissues in the ¯ounders and are reported too. On an average 3.0% of all ¯ounders examined (>20 cm) were aected by preneoplastic/ neoplastic liver lesions. The prevalence is signi®cantly increasing with increasing size of ®sh. Of specimens >30 cm 10.1% were aected and, at one site the disease prevalence in female ¯ounder >30 cm was 31.5%, which must be considered strikingly high. The site not immediately in¯uenced by human activities but in closest contact with the main Baltic Sea, had the highest lesion prevalences. The levels of hydrophobic DNA-adducts in the liver of ¯ounder in the area are low which indicates that, for example, polycyclic aromatic hydrocarbon compounds are not responsible for the high prevalences of proliferative lesions in ¯ounder in this area. PII: S0141-1136(00)00207-5