Use of fish in vitro hepatocyte assays to detect multi-endpoint toxicity in Slovenian river sediments.
There is an increasing demand for rapid, sensitive and robust methods for toxicity testing of single chemicals, complex mixtures and environmental samples. The objective of this work was to validate and use a primary culture of rainbow trout (Oncorhynchus mykiss) hepatocytes as a multi-endpoint in vitro bioassay for toxicity characterisation of river sediments from four areas of the Sava and Krupa Rivers (Slovenia). The endpoints were chosen to encompass acute toxicity (cytotoxicity) as well as sub-lethal biomarker and effect endpoints such as metabolic inhibition, DNA damage (Fast Micromethod), endocrine disruption (estrogenicity), and 7-ethoxyresorufin O-deethylase (EROD) activity. Results from these studies show that the primary hepatocyte culture was able to successfully detect effects of single model chemicals in all endpoints analysed. Furthermore, the bioassays were also able to discriminate between contaminated and less contaminated sediments for a number of endpoints such as cytotoxicity, metabolic inhibition and induction of EROD activity, although no increase in DNA damage and estrogenicity was observed above background at any site. The present study shows that primary fish hepatocytes may be used to determine multiple mechanisms of toxic action and that a holistic assessment of effects may improve our understanding of cellular toxicity of complex mixtures such as sediments.
Tollefsen
Knut-Erik
KE
Norwegian Institute for Water Research (NIVA), Kjelsaas, Oslo, Norway. knut.erik.tollefsen@niva.no
Bratsberg
Erling
E
Bøyum
Olav
O
Finne
Eivind Farmen
EF
Gregersen
Inger Katharina
IK
Hegseth
Marit
M
Sandberg
Cecilie
C
Hylland
Ketil
K
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity; D000818:Animals; D001681:Biological Assay / Q000379:methods; D002478:Cells, Cultured; D019363:Cytochrome P-450 CYP1A1 / Q000032:analysis; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000379:methods*; D019015:Geologic Sediments / Q000737:chemistry; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects*; D007928:Lethal Dose 50; D017686:Oncorhynchus mykiss / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D010636:Phenols / Q000633:toxicity; D015203:Reproducibility of Results; D017524:Slovenia; D018675:Toxicity Tests / Q000379:methods*; D014818:Vitellogenesis / Q000187:drug effects
D010636:Phenols; D015112:4-Nitroquinoline-1-oxide; C025256:nonylphenol; D019363:Cytochrome P-450 CYP1A1