Strain difference in the micronucleus test

The Collaborative Study Group for the Micronucleus Test, a task group of the Environmental Mutagen Society of Japan, has earlier addressed the question of sex difference as a source of variation in the micronucleus test. Strain difference, another issue in test protocols requiring urgent clarification, was selected as the subject of the second study. Male mice of strains Slc: ddY (ddY), CRJ: CD-1(ICR) (CD-1), Slc: BDF₁ (BDF₁), and ms: Hal (ms) were treated with 6 different chemicals chosen from various classes of micronucleus inducers: colchicine, 7,12-dimethylbenz[a]anthracene, ethyl methanesulfonate, N-ethyl-N-nitrosourea, 6-mercaptopurine, and potassium chromate. All 4 strains gave positive results with all 6 chemicals, although ms tended to show the highest responses. ddY and CD-1 were low responders, while BDF₁ was intermediate between ms and the other two. Although ms seemed superior to the other strains, its high responses became manifest mostly at high dose levels. ms was not always the most sensitive strain; it responded moderately to ethyl methanesulfonate. Also the background level of micronucleated polychromatic erythrocytes was the highest in ms, but this did not explain the apparent high sensitivity of this strain. Despite the strain differences, it can be concluded that any of the other strains used seems to suffice as a tester for the micronucleus test.     

Thirty compounds tested in the Drosophila wing spot test

The Drosophila wing somatic mutation and recombination test (SMART) was evaluated for its suitability in genotoxicity screening by testing 30 chemicals. Of the 2 crosses used, the mwh-flr3 cross turned out to be more convenient than the previously used mwh-flr cross. Based on the experience gained with both acute exposures and chronic exposures of different duration, we suggest that the optimal strategy in genotoxicity screening is to start with chronic exposure of 3-day-old larvae for 48 h (that is, until pupation). Only for unstable compounds and very volatile compounds and gases are acute treatments, including inhalation, recommended. In general, a qualitative evaluation of the genotoxicity of a compound in the wing assay is possible with as few as 1-2 different exposure concentrations. A more quantitative evaluation of genotoxicity, based upon dose-response data, can often be achieved with as few as 3-4 concentrations. The results reported here were obtained in 2 different laboratories, demonstrating that the wing spot test is easily transferable to other laboratories. The experience gained indicates that the assay has now been developed to an extent that a coordinated international comparative validation study is desirable.     

Possible recombinogenic effect of caprolactam in the mammalian spot test

Tests of caprolactam in the mouse spot test showed that treatment with this compound increased the frequency of color spots among animals treated in utero. The nature of these spots suggests that caprolactam may induce spots through the induction of mitotic recombination.     

Genotoxicity of four herbicides in the Drosophila wing spot test.

The herbicides alachlor, atrazine, maleic hydrazide and paraquat were evaluated for genotoxicity in the Drosophila melanogaster wing spot test. Third-instar larvae trans-heterozygous for two recessive mutations of wing trichomes, multiple wing hairs (mwh) and flare (flr3), were treated by chronic feeding with different concentrations of the four herbicides. Feeding ended with pupation of the surviving larvae. The genotoxic effects were determined from the appearance of clones of cells with mwh, flr3 or mwh-flr3 phenotypes. Exposure to maleic hydrazide resulted in a significant increase in the frequency of the three categories of spots recorded (small single, large single and twin spots) in a dose-related fashion. Exposure to alachlor induced significant increases in both small and total spots at the four concentrations assayed and in the frequency of twin spots at the highest concentration tested (10 mM). Atrazine and paraquat also induced significant increases in both small and total spots at three of the four concentrations tested, without indication of a direct dose-effect relationship.     

Genotoxicity testing of five herbicides in the Drosophila wing spot test

Four triazine herbicides: amitrole, metribuzin, prometryn and terbutryn, and the bipyridal compound diquat dibromide have been evaluated for genotoxicity in the wing somatic mutation and recombination test of Drosophila melanogaster, following standard procedures. Third-instar larvae trans-heterozygous for the third chromosome recessive markers multiple wing hairs (mwh) and flare-3 (flr(3)) were chronically fed with different concentrations of the test compounds. Feeding ended with pupation of the surviving larvae. Genetic changes induced in somatic cells of the wing's imaginal discs lead to the formation of mutant clones on the wing blade. Point mutation, chromosome breakage and mitotic recombination produce single spots; while twin spots are produced only by mitotic recombination. Exposure to 0.5 mM and 1 mM of amitrole clearly increased the frequency of small single, large single and total spots. Terbutryn, at the concentration of 5 mM, induced a slight increase in the frequency of small single and total spots, but this result could be false positive. The other three herbicides tested did not show any genotoxic effect. When heterozygous larvae for mwh and the multiple inverted TM3 balancer chromosomes were treated, significant increases in the frequency of mutant spots were only detected for amitrole. The observed spot frequencies were lower than those found in mwh/flr(3)50%) of the total spot induction was due to mitotic recombination.