S-XllI : Identification and evaluation of environmental mutagens, ecogenotox ico!ogy SI19

The METAFER2 Automated Metaphase Finder (MetaSystems, Sandhausen,Germany) is based on an IBM-AT compatible computer equipped with a bi­nary image analysis board, a light microscope with motorized scanning stageand autofocus, and a high resolution CCD camera. Cytogenetics analysis isperformed in a two step process . The METAFER2 metaphase finder scansa slide frame by frame at a 16 x magnification using predefined class ifiersfor e.g. human lymphocytes or Chinese hamster metaphase. Chromosomeanalysis is then performed using the 100 x magnification oil objective.In order to demonstrate that METAFER2 allows an unbiased selection ofmetaphases, human lymphocytes or Chinese hamster permanent ceUs prepa­rations were automatically or manually scanned . The comparison for slideswith a high percentage of metaphases containing structural chromosomeaberrations (mult iple aberrant cells were also included in the evaluation)as weU as numerical chromosome changes (polyploid and endoreduplicatedceUs were evaluated) shows that METAFER2 does not detect the non­aberrant metaphases at a higher or lower probability than the aberrantones. Since random selection of metaphases is neither influenced by variousstructural chromosome aberrations nor by increased frequencies of numericalchromosome changes, the METAFER2 system can be used to performroutine chromosomal aberration assays using human lymphocytes or Chinesehamster cells,

Keyword(s): Automated metaphase finding; Chromosome aberration assays;Image analysis

Removal of residual chlorine: For removal of residual chlorine, Na2S203as reducing agent of chlorine and nitrogen as vaporizing gas were used. Themutagenic activity of water sample treated with Na2S203, not with nitrogen,decreased.

Sample storage ; Three storage methods were compared After collectingof water samples, they were kept at 4', ·scr for I month. Higher mutagenicactivity wasobtained in the case of 4' compared with those kept at -80' andcollected before use.

Eluant of adsorbates; Dichlorornethane and ethyl acetate were used asextracting solvent of mutagenic substances . These solvents showed similarextracting efficiency and mutagenic results.

When the water supplying source had troubles such as drought or anremarkable increase of algae, the mutagenic activity ofchlorinated tap waterwas significantly high value.

These data shows that how to remove chlorine and how to storage thechlorinated water samples are essential factors for determination of themutagenic activity.

Keyword(s); Chlorinated by-products; Drinking water; Ames test

Ip XIII.ll1l Mutagenic halogenated furanones occurring In tapwater

Boiena Chlopkiewicz. Drug Institute. 30/34 ChelmskaStr; 00-725 Wanaw,Poland

Adriamyc in and bleomycin are widely used as effective antineoplastic drugsin the treatment of number of human cancer. The mechanism behindthe cytostatic activity of adriamycin and bleomycin is not yet completelyunderstood, although, a number of cytotoxic and genotoxic effects of drugshave been described . Various investigators have proposed that the antitumoror genotoxic activity of adriamycin resulted from drug-induced free radicalformation . Genotoxic activity of adriamycin and bleomycin in embryo cellsfrom mice differ in anti-oxidant enzymes activity was investigated. Thecatalase activity in cultured ill vitroembryo ceUsofC3H mice was 2-fold andsuperoxide dismutase 3-fold higher than of CS7BUI0 mice. For genotoxicityevaluation, the micronucleus test In vitro was used. The results obtainedindicated that frequency of micronucleated cells in untreated C3H cultureswas higher than in CS7BUIO cell cultures . The increase in micronucleiformation after treatment with adriamycin and bleomycin was higher inCS7BUIO than in C3H cells as compare with micronuclei in untreatedcultures. The higher frequency of micronucleated cells in treated towardsuntreated CS7BUIO than C3H cell cultures may be caused by lower activityof anti-oxidant enzymes in CS7BUI0 cells. It may suggests that DNAdamage caused by adriamycin and bleomycin resulted from action of activeoxygen species.

Ip XIII.I091 III vitroevaluation of genotoxlc activity of adrlamyclnand bleomycin In mouse embryo cells which differ Incatalase and superoxlde dlsmutase actIvity

Isabel Ramos, Maia Lloveras, Maria-Pilar Marco, Angel Messeguer. Dpt. ofBiologicalOrganic Chemistry. CID-CSIC. J. Girona. 18. 08034 Barcelona.Spa",

The occurrence of halogenated futanones in drinking water is a matter ofconcern due to the mutagenic ity exhibited by these compounds. The most im­portant representat ive of this family of toxins is 3-chloro.4-(dichlorornethyl)­S-hydroxy-2(SH)-furanone, a compound known also as MX. This furanonehas been widely detected in samples from chlorinated drinking and humicwater. MX turned out to be one of the most potent direct acting mutagensever tested in the Ames assay and it has been found to induce genotoxicityin numerous ill vitro and ill vivo assays (I). On the other hand, in chlorinedisinfected drinking water from a raw water with a high brom ide content,bromohydroxyfuranones could also be present (2). Consequently, levelsof halogenated furanone derivatives should be adequately mon itored andminimized.

In the present communication the rigorous characterisation of differenthalogenated furanones containing bromine atoms will be presented. In addi­tion. preliminary results on the mutagen ic activity and potential occurrenceof these compounds in water samples will be also discussed.

Keyword(s): Mutagens; Halogenated furanones

(I) Kronberg, L.; Franzen, R. Eroiron. Sci. Technol..• 1993,27. 1811-1818.(2) Horth, H. J. Fr. Hydrol.• 1990,21, I3S-14S.

Keyword(s): Adriamycin; Bleomycin; Anti-oxidant enzymes

Ip XIII.HOI Mutagenic activity of chlorinated tap water

Mitsulco Kato, Haruko Saito, Shin'ichi lsoda, Noboru Nagaolca. YofcahamaCity lnstit: ofHealth, ]35. Yokohama, Japan

Recently, reports on mutagenic substances in contained tap water are in­creased in number. In this work, the pretreatment techniques for isolation ofmutagens followed by the Ames test and the change of mutagenic activityof several municipal drinking water were studied .

Each water sample was passed through an XAD-2 resin column andadsorbates were eluted with a solvent. The eluate was concentrated andthen submitted to the Ames test.