ALTEX :: Alternatives to Animal Experiments

1995, Volume 4

Toxicity Assessment of Waste Water Samples with Fish Cell Lines

Markus Schulz1, Bettina Lewald1, Martin Kohlpoth2, Brigitte Rusche2, Katrin H.J. Lorenz3, Eckehardt Unruh3, Peter-D. Hansen3, Herbert G. Miltenburger1

1Technische Hochschule Darmstadt, Laboratorium für Mutagenitätsprüfung (LMP), D-Darmstadt; 2Akademie für Tierschutz, D-Neubiberg; 3Technische Universität Berlin, Institut für Ökologie und Biologie, Ökotoxikologie, D-Berlin

SUMMARY

An in vitro cytotoxicity assay with fish cell lines was established in order to reduce the number of animals used in fish tests with the golden orfe (leuciscus idus melanotus). In three co-ordinated working laboratories the toxicity of seven chemicals was determined according to a standardized protocol using the permanent cell lines D11 and RTG-2. There was an almost linear correlation (r=0.99) between the IC50 values after neutral red uptake and crystal violet staining of the chemically treated RTG-2 cells. These cells were more sensitive than D11 cells (r=0.93) with regard to chemically induced cytotoxicity.

The test protocol was adapted for the testing of waste water (ten samples). Two samples induced cytotoxic effects in RTG-2 cells while in none of the three laboratories the same samples were found cytotoxic in D11 cells. The data with the seven chemicals and the still limited number of waste water samples (10) lend strong support to the assumption that the fish cell line RTG-2 can be regarded as well suited for detecting toxic properties of waste water.

Keywords: in vitro cytotoxicity, fish cell line, RTG-2, D11, neutral red assay, waste water samples