Evaluation of the rat bone marrow and peripheral blood micronucleus test using monocrotaline.

The present study was performed as part of the 9th collaborative trial of the CSGMT (Collaborative Study Group for the Micronucleus Test) to evaluate the performance of the rat micronucleus test in detection of clastogenic agents. Temporary fluorescent staining using acridine orange (AO) was compared with permanent staining using a modified Feulgen technique for bone marrow and Giemsa for blood. The Feulgen and AO methods were both found to be suitable for demonstrating micronucleus induction in rat bone marrow following oral administration of monocrotaline and cyclophosphamide. Induction of micronuclei was also shown using rat blood. The simple AO supravital method described produced uniformly stained preparations which were extremely easy to assess and interpret. The AO method was preferred to Giemsa for analysis of blood, but it was essential to analyse samples within 4 days of collection to avoid excessive deterioration of cells. Because of the relatively low number of micronucleated cells found in rat blood, it is suggested that examination of a larger number of cells may be necessary to achieve a similar sensitivity to the bone marrow assay for routine screening.

Assessment of the potential in vivo genotoxicity of three trihalomethanes: chlorodibromomethane, bromodichloromethane and bromoform.

Chlorination of drinking water results in the formation of chlorodibromomethane, bromodichloromethane and bromoform. These trihalomethanes have all shown evidence of genotoxicity in bacterial and mammalian cell systems in vitro and some evidence of carcinogenicity in rodents. Chlorodibromomethane and bromodichloromethane have previously been tested in the mouse micronucleus test and did not induce chromosome damage, but results from two previous micronucleus tests on bromoform are somewhat contradictory. In the present study, bromoform was tested in the mouse bone marrow micronucleus test in order to reassess the response in this system; all three compounds were evaluated using the rat liver unscheduled DNA synthesis test. Trihalomethanes are well absorbed by the oral route which was selected for this study as being that most relevant to humans. Bromoform did not induce micronuclei in mouse bone marrow, and chlorodibromomethane, bromodichloromethane and bromoform did not cause unscheduled DNA synthesis in rat liver. These trihalomethanes have not shown any evidence of genotoxicity in vivo and are most unlikely to have any significant genotoxic activity in mammals. Their mode of action as rodent carcinogens remains unexplained.

Assessment of the potential in vivo genotoxicity of fluoranthene.

Fluoranthene is a ubiquitous environmental pollutant. Although fluoranthene is mutagenic in bacterial and mammalian in vitro cell systems following metabolic activation by rat liver fraction, information on in vivo mutagenicity is lacking and studies on tumour initiating activity in mice are equivocal. In the present study, the potential genetic hazard to man was assessed using the mouse bone marrow micronucleus and rat liver unscheduled DNA synthesis test systems. Fluoranthene did not show any evidence of genotoxicity in either of the in vivo assays following acute oral administration at levels of up to 2000 mg/kg b.w.

Genotoxicity testing of pidotimod in vitro and in vivo.

The mutagenic potential of pidotimod ((R)-3-[(S)-(5-oxo-2- pyrrolidinyl)carbonyl]-thiazolidine-4-carboxylic acid, PGT/1A, CAS 121808-62-6) was assessed in a series of five assays designed to measure gene mutation, chromosomal damage and primary DNA damage. All tests were carried out in accordance with appropriate EEC and OECD Guidelines. No indications of mutagenic potential were observed in any of the assays.

Evaluation of the mutagenicity of a-dihydroergocryptine in vitro and in vivo.

The mutagenic potential of the ergot alkaloid a-dihydroergocryptine (a-DEC, CAS 14271-05-7) was assessed in a series of 5 assays designed to measure gene mutation, chromosomal damage and primary DNA damage. All tests were carried out in accordance with appropriate EEC and OECD Guidelines. No indications of mutagenic potential were observed in any of the assays.

Assessment of the genotoxic potential of Caramel Colour I in four short-term tests.

A battery of three short-term tests in vitro and one in vivo was used to determine the genotoxicity of Caramel Colour I. The results of the bacterial mutation assay, using five strains of Salmonella typhimurium, and the mouse micronucleus assay in vivo showed no evidence of genotoxic activity. Results from both the cytogenetics assay in vitro, using CHO cells, and the mouse lymphoma assay indicated that there was some genotoxic activity associated with Caramel Colour I but only in the absence of S-9 and at very high dose levels.