Genotoxic and Ecotoxic Effects of Groundwaters and Their Relation to Routinely Measured Chemical Parameters CHRISTOPH HELMA,* ,† PETER ECKL, ‡ EVA GOTTMANN, † FEKADU KASSIE, † WOLFGANG RODINGER, § HANS STEINKELLNER, † CHRISTIAN WINDPASSINGER, † ROLF SCHULTE-HERMANN, † AND SIEGFRIED KNASMU ¨ LLER † Institute for Tumor Biology, Cancer Research, University of Vienna, Borschkegasse 8a, A-1090 Vienna, Austria, Institute for Genetics and Developm ental Biology, University of Salzburg, Hellbrunnerstrasse 34, A-5020 Salzburg, Austria, Institute for Water Quality, Federal Agency for Water Managem ent, Schiffm u ¨ hlenstrasse 120, A-1220 Vienna, Austria The primary aim of the present investigation was to study possible adverse effects of groundwater from an aquifer south of Austria’s capital, Vienna, and to relate these toxicological effects to routinely measured physical/chemical parameters. Fourty-three water samples were tested for genotoxic and ecotoxic effects. For genotoxicity testing the Salmonella/microsome assay, the micronucleus test with primary rat hepatocytes and micronucleus tests with plants (Tradescantia, Vicia faba)were used. In ecotoxicity tests, algae (Selenastrum capricornutum), water cress (Le- pidium sativum), and water flea (Dapnia magna) were studied as target organisms. In genotoxicity assays, 10 samples (23%) gave a weak positive response with a single end point, but only one sample (2%) was genotoxic in three different test systems. Thirty-six samples (86%)caused adverse effects in ecotoxicity assays. Plants (algae and water cress) were more sensitive than daphnie. No correlations between toxic effects and physical/chemical parameters were detected. The genotoxicity experiments indicate presently a low risk from genotoxic compounds. The ecotoxic (especially phytotoxic) properties of many water samples raise concern about their suitability for irrigation purposes. The lacking correlation between results from toxicity tests and physical/chemical data indicates that it is presently impossible to predict toxic properties from routine physical/chemical measurements with a sufficient level of safety. It is therefore important to include biological toxicity assays in groundwater monitoring programs. Introduction Groundwater is an important source for drinking and irrigation watersupplies. The contamination ofaquiferswith substances originating from agricultural and industrial activities or waste disposal has raised much public concern during the last years (1). Scientific and regulatoryefforts are presentlyfocused on the identification and quantification of single chemicals and summary parameters that are consid- ered to be harmful for man or the environment. This approach is based on the assumptions (a) that all harmful compounds are known and accessible to chemical analysis and (b) that the compounds present in the environment interact accordingto a certain model(e.g.,additivity).Areview ofthe present literature (2)reveals that onlya minor fraction ofthe organicsubstancespresent in waterhasbeen identified. Antagonistic and synergistic effects of water contaminants are not predictable a priori, and the toxicological properties of many water contaminants are unknown. Therefore,it was proposed to use biologicaltoxicitytests, which were initially developed for the safety evaluation of chemicals, directly for environmental samples. Although manyreports exist on toxic properties ofaqueous media (for reviews, see refs 3-6), only a very few studies have been published about adverse toxicologicaleffectsofgroundwater (7-12). The primary objective of this study was to investigate if DNA-damaging (genotoxic) compounds are present in one ofthe largest groundwater reservoirsofAustria located south ofthe capital Vienna and to clarify ifcontaminated drinking water might be responsible for the slightly elevated cancer rate observed in this region. It is well-known that the majority ofcarcinogensactbygenotoxicmechanisms(13). Therefore, short-term genotoxicitytests are used to identifycarcinogens prior to long-term animal studies. We applied a battery of genotoxicity assays with different target organisms, namely, the Salm onella/microsome assay(14),the micronucleustest with primary rat hepatocytes (15), and micronucleus assays with plants (Tradescantia (16)and Vicia faba (17)) to detect genotoxic and probably carcinogenic properties of ground- water from the investigated area. These tests were supplemented with ecotoxicityassays to identifyadverse effects toward plants and invertebrates that may result from agricultural use of the groundwater (e.g., irrigation). Standardized ecotoxicity tests were conducted with algae (Selenastrum capricornutum ;(18), water cress (Lepidium sativum ; 19), and water flea (Daphnia magna ; 20). Finally, we compared the results from toxicity tests with physical/chemicalparametersroutinelymeasured according to Austrian Federal regulations (21). The primary aim was to identify compounds responsible for toxic effects. The second aim was to clarify if it is possible to substitute expensive toxicity assays with the measurement of selected chemical parameters. Experimental Protocol Sample Sources. From 1994 to 1996, 43 samples were taken from 26 locations in the investigation area, some of them (locations 3, 12, 17, 22, 29, 35, and 40) repeatedly in different years. The sample codes used in this paper are composed bya numberindicatingthe location followed bythe sampling year (e.g., 3-94 location 3 in 1994). The majority of samples were taken from wells whose water is currently not used as human drinking water (locations 3-5, 12, 13, 16-19, 22, 24, 29,39,and 109). In addition,five localwater works (locations 35, 101, 102, 104, and 105) and one reclamation plant for groundwater contaminated by a landfill (location 40) were included in this study. At locations 35, 105, and 40, different *To whom correspondence should be addressed. Phone: +43- 1-401 54-226; fax: +43-1-406 07 90; e-mail: Christoph.Helma@univie.ac.at. † University of Vienna. ‡ University of Salzburg. § Federal Agency for Water Management. Environ. Sci. Technol. 1998, 32, 1799-1805 S0013-936X(97)01049-3 CCC: $15.00  1998 American Chemical Society VOL. 32, NO. 12, 1998 / ENVIRONMENTAL SCIENCE & TECHNOLOGY 9 1799 Published on Web 05/09/1998