Dominant Lethal Test Research Articles

Toxicological, medical and industrial hygiene aspects of glutaraldehyde with particular reference to its biocidal use in cold sterilization procedures

Aqueous solutions of > or =5% glutaraldehyde (GA) are of moderate acute peroral toxicity and those of < or =2% are of slight toxicity. By single sustained skin contact, aqueous GA solutions of > or =45% are of moderate acute percutaneous toxicity, those of 25% are of slight toxicity and those of </=15% do not present an acute percutaneous hazard. Vapor generated at ambient temperature may cause sensory irritant effects to the eye and respiratory tract, but not acute respiratory tract injury. The 50% decrease in respiratory rate (rd(50)) is 13.86 ppm. A 0.1% solution of GA is not irritating to the eye; the threshold for conjunctival irritation is 0.2% and for corneal injury it is 1.0%. Eye injury is moderate at 2% and severe at > or =5%. Primary skin irritation depends on the duration and contact site, occlusion and solvent. By sustained contact, the threshold for skin irritation is 1%, above which erythema and edema are dose related. With 45% and higher, skin corrosion may occur. There is a low incidence of skin sensitizing reactions, with an eliciting threshold of 0.5% aqueous GA. However, GA is neither phototoxic nor photosensitizing. Subchronic repeated exposure studies by the peroral route show only renal physiological compensatory effects, secondary to reduced water consumption. Repeated skin contact shows only minor skin irritant effects without systemic toxicity. By subchronic vapor exposure, effects are limited to the nasal mucosa at 1.0 ppm, with a no-effect concentration generally at 0.1 ppm. There is no evidence for systemic target organ or tissue toxicity by subchronic repeated exposure by any route. A chronic drinking water study showed an apparent increase, in females only, of large granular cell lymphocytic leukemia but this was not dosage related. This is most likely the result of a modifying effect on the factor(s) responsible for the expression of this commonly occurring rat neoplasm. A chronic (2-year) inhalation toxicity/oncogenicity study showed inflammatory changes in the anterior nasal cavity but no neoplasms or systemic toxicity. In vitro genotoxicity studies--bacterial mutagenicity, forward gene mutation (HGPRT and TK loci), sister chromatid exchange, chromosome aberration, UDS and DNA repair tests--have given variable results, ranging from no effect through to weak positive. In vivo genotoxicity studies--micronucleus, chromosome aberration, dominant lethal and Drosophila tests--generally have shown no activity but one mouse intraperitoneal study showed bone marrow cell chromosome aberrations. Developmental toxicity studies show GA not to be teratogenic, and a two-generation study showed no adverse reproductive effects. Percutaneous pharmacokinetic studies showed low skin penetration, with lowest values measured in vitro in rats and human skin. Overexposure of humans produces typical sensory irritant effects on the eye, skin and respiratory tract. Some reports have described an asthmatic-like reaction by overexposure to GA vapor. In most cases this resembles reactive airways dysfunction syndrome, and the role of immune mechanisms is uncertain. Local mucosal effects may occur if medical instruments or endoscopes are not adequately decontaminated. Protection of individuals from the potential adverse effects of GA exposure requires that there be adequate protection of the skin, eyes and respiratory tract. The airborne concentration of GA vapor should be kept below the recommended safe exposure level (e.g. the threshold limit value) by the use of engineering controls. Those who work with GA should, through a training program, be aware of the properties of GA, its potential adverse effects, how to handle the material safely and how to deal with accidental situations involving GA. If effects develop in exposed workers, the reasons should be determined immediately and corrective methods initiated. (c) 2001 John Wiley & Sons, Ltd.

On Bayesian analyses and finite mixtures for proportions

When the results of biological experiments are tested for a possible difference between treatment and control groups, the inference is only valid if based upon a model that fits the experimental results satisfactorily. In dominant-lethal testing, foetal death has previously been assumed to follow a variety of models, including a Poisson, Binomial, Beta-binomial and various mixture models. However, discriminating between models has always been a particularly difficult problem. In this paper, we consider the data from 6 separate dominant-lethal assay experiments and discriminate between the competing models which could be used to describe them. We adopt a Bayesian approach and illustrate how a variety of different models may be considered, using Markov chain Monte Carlo (MCMC) simulation techniques and comparing the results with the corresponding maximum likelihood analyses. We present an auxiliary variable method for determining the probability that any particular data cell is assigned to a given component in a mixture and we illustrate the value of this approach. Finally, we show how the Bayesian approach provides a natural and unique perspective on the model selection problem via reversible jump MCMC and illustrate how probabilities associated with each of the different models may be calculated for each data set. In terms of estimation we show how, by averaging over the different models, we obtain reliable and robust inference for any statistic of interest.

Bleomycin, unlike other male-mouse mutagens, is most effective in spermatogonia, inducing primarily deletions

Dominant-lethal tests [P.D. Sudman, J.C. Rutledge, J.B. Bishop, W.M. Generoso, Bleomycin: female-specific dominant lethal effects in mice, Mutat. Res. 296 (1992) 205–217] had suggested that Bleomycin sulfate (Blenoxane), BLM, might be a female-specific mutagen. While confirming that BLM is indeed a powerful inducer of dominant-lethal mutations in females that fails to induce such mutations in postspermatogonial stages of males, we have shown in a specific-locus test that BLM is, in fact, mutagenic in males. This mutagenicity, however, is restricted to spermatogonia (stem-cell and differentiating stages), for which the specific-locus mutation rate differed significantly ( P<0.008) from the historical control rate. In treated groups, dominant mutations, also, originated only in spermatogonia. With regard to mutation frequencies, this germ-cell-stage pattern is different from that for radiation and for any other chemical studied to date, except ethylnitrosourea (ENU). However, the nature of the spermatogonial specific-locus mutations differentiates BLM from ENU as well, because BLM induced primarily (or, perhaps, exclusively) multilocus deletions. Heretofore, no chemical that induced specific-locus mutations in spermatogonia did not also induce specific-locus as well as dominant-lethal mutations in postspermatogonial stages, making the dominant lethal test, up till now, predictive of male mutagenicity in general. The BLM results now demonstrate that there are chemicals that can induce specific-locus mutations in spermatogonia without testing positive in postspermatogonial stages. Thus, BLM, while not female-specific, is unique, (a) in its germ-cell-stage specificity in males, and (b) in inducing a type of mutation (deletions) that is atypical for the responding germ-cell stages (spermatogonia).

Mutagenic evaluation of deltamethrin using rodent dominant lethal assay

The dominant lethal test was used to analyse the mutagenic potential of deltamethrin, a synthetic pyrethroid insecticide, in Swiss albino mice. In the treated series, the animals were exposed orally to three different doses (0.36, 0.72 and 1.08 mg/kg body weight) of deltamethrin dissolved in corn oil. Following the treatment, each male of control, as well as of the treated series, was mated with untreated females, every week for a period of 6 weeks. All mated females were sacrificed on the 13th day of separation and their ovaries and uterus were examined. The results revealed that deltamethrin treatment did not impair the mating capacity and fertility of Swiss albino mice. Mutagenic index, pre- and post-implantation losses were assessed. No significant pre-implantation losses were observed either weekly or averagely. Post-implantation losses were observed at medium and high doses of deltamethrin. A slight increase in dominant lethal mutation rate was observed by increasing doses of deltamethrin in early weeks but decreased in later weeks, so an apparent dose response was not observed.

Recommendations for statistical designs of in vivo mutagenicity tests with regard to subsequent statistical analysis

A workshop was held on September 13 and 14, 1993, at the GSF, Neuherberg, Germany, to start a discussion of experimental design and statistical analysis issues for three in vivo mutagenicity test systems, the micronucleus test in mouse bone marrow/peripheral blood, the chromosomal aberration tests in mouse bone marrow/differentiating spermatogonia, and the mouse dominant lethal test. The discussion has now come to conclusions which we would like to make generally known. Rather than dwell upon specific statistical tests which could be used for data analysis, serious consideration was given to test design. However, the test design, its power of detecting a given increase of adverse effects and the test statistics are interrelated. Detailed analyses of historical negative control data led to important recommendations for each test system. Concerning the statistical sensitivity parameters, a type I error of 0.05 (one tailed), a type II error of 0.20 and a dose related increase of twice the background (negative control) frequencies were generally adopted. It was recommended that sufficient observations (cells, implants) be planned for each analysis unit (animal) so that at least one adverse outcome (micronucleus, aberrant cell, dead implant) would likely be observed. The treated animal was the smallest unit of analysis allowed. On the basis of these general consideration the sample size was determined for each of the three assays. A minimum of 2000 immature erythrocytes/animal should be scored for micronuclei from each of at least 4 animals in each comparison group in the micronucleus assays. A minimum of 200 cells should be scored for chromosomal aberrations from each of at least 5 animals in each comparison group in the aberration assays. In the dominant lethal test, a minimum of 400 implants (40–50 pregnant females) are required per dose group for each mating period. The analysis unit for the dominant lethal test would be the treated male unless the background frequency of dead implants (DI) is so low that multiple males would need to be integrated to meet the minimum observation of one adverse outcome (DI) per analysis unit. A three-step strategy of data analysis was proposed for the cytogenetic assays. Use of negative historical controls was allowed in certain circumstances for interpretation of results from micronucleus tests and chromosomal aberration tests.

Dominant lethal test with rats exposed to 1,3-dichloropropene by inhalation.

The potential of 1,3-dichloropropene (1,3-DCP) to induce dominant lethal mutations in the germ cells of male CD rats following inhalation exposure was investigated. Groups of 11-week-old males (30 animals/group) were exposed to 1,3-DCP vapors by inhalation at targeted concentrations of 0 (negative control), 10, 60, and 150 ppm for 10 weeks (6 hr/day, 7 days/week). An additional group of 30 males (designated the pairfed group) was kept on dietary restriction for 10 weeks. This group served as a control for any effects of decreased feed consumption and the associated body weight loss on the dominant lethal indices in the males exposed to 1,3-DCP. At the termination of the exposures, each male was cohoused with naive adult virgin CD females for two consecutive mating trials (1 week/trial, 2 females/male). Females were necropsied 13 days after the conclusion of each weekly mating trial and the number of corpora lutea, live implantations, and resorptions were determined. There were no statistically significant increases in either the pre- or postimplantation embryonic/fetal loss in females mated with 1,3-DCP-exposed males relative to controls at any weekly mating period. Based on these results, it can be concluded that 1,3-DCP is not mutagenic to the male germ cells of CD rats at exposure levels < or = 150 ppm, the highest concentration tested.

Dose response study for 1,3-butadiene-induced dominant lethal mutations and heritable translocations in germs cells of male mice

Butadiene (BD) and its metabolites have extensively been studied in the EU sponsored research project `Multi-endpoint analysis of genetic damage induced by 1,3-butadiene and its major metabolites'. Within this project a dominant lethal test and a heritable translocation test were performed with male mice to study the dose-response relationships for the respective endpoints. BD concentrations of 130 and 500 ppm were tested in the dominant lethal assay by exposing male mice on 6 h/day for five consecutive days resulting in doses of 3900 and 15 000 ppmh, respectively. Males were mated for four consecutive weeks at a ratio of 1:2 to untreated females. A positive dominant lethal effect was observed in the first mating week in the experiment with 15 000 ppmh but no dominant lethality was found with the lower dose of 3900 ppmh. The present dominant lethal data have to be viewed together with the data already published for a BD dose of 39 000 ppmh (1300 ppm at 6 h/day on 5 consecutive days) [1]. The main difference between results with the highest and the middle dose is that mating weeks one and two (sperm and late spermatids) showed an effect at 39 000 ppmh while only mating week one (sperm) showed an effect at 15 000 ppmh. In the heritable translocation assay, males mice were exposed with a BD dose of 15 000 ppmh and mated for one week to untreated females. Among 434 F 1 offspring, we found 5 translocation carriers (1.15% vs. 0.05% in the historical control, p<0.001). In the previous heritable translocation experiment with a BD dose of 39 000 ppmh of DB exposure, 2.7% of the offspring carried a reciprocal translocation [2]. These data can be used for quantification of genetic risk. The dose response for BD-induced heritable translocations in sperm and late spermatids of mice was linear ( Y=0.05+6.9×10 −5 X) and a doubling dose of 725 ppmh could be calculated.

Assessment of mutagenic potential of thiram

Thiram is a widely used dithiocarbamate fungicide. In this study, the mutagenicity of thiram was investigated using the micronucleus and dominant lethal tests in Swiss albino mice. A single ip injection of 100 mg thiram/kg body weight, which is the maximum tolerated dose (MTD), significantly induced micronucleus formation in bone marrow cells after 30 and 48 hr of exposure; 50% and 25% of the MTD also induced micronucleus formation after the above time periods. A significant number of dead implants were induced when thiram was given to male mice in the diet at 10% of the oral LD 50 during the whole spermatogenesis cycle (8 wk); this post-implantation loss indicates a dominant lethal mutation.

Mammalian germ cell mutagenicity of ENU, IPMS and MMS, chemicals selected for a transgenic mouse collaborative study

A collaborative study to systematically assess transgenic mouse mutation assays as screens for germ cell mutagens has been conducted. Three male mouse germ cell mutagens (ENU, iPMS and MMS) were selected for testing. This paper provides a brief review of the effects reported for those 3 chemicals in the most commonly used non-transgenic germ cell mutagenicity assays, namely the dominant lethal, heritable translocation, and specific locus tests. Additionally, information on the DNA reactivity and the molecular nature of mutations induced by these chemicals is summarized.

5554377 Non-toxic animal repellent : Abraham Carl J Great Neck, NY, United States

The soluble organic fraction of diesel particulate emissions was tested for physiological and genotoxic effects on the parasitoid wasp, Bracon hebetor. Adult female survival after topical treatment was significantly decreased at the higher concentrations of SOF. Daily egg production, a sensitive assay for genotoxic and physiological effects, was not significantly affected at any concentration. Egg hatchability on Days 1–5, representing vitellogenic oocytes at the time of treatment, was decreased, but this effect lacked consistency and dose dependence. An extensive dominant lethal test was performed with negative results. Possible explanations for these findings and comparisons with in vitro tests such as the Ames test and sister chromatid exchange are discussed.

Mutagenicity of anticancer drugs in mammalian germ cells

The evidence for mammalian germ cell mutagenicity induced by anticancer drugs is summarized. Primary attention is paid to the three major mouse germ cell mutagenicity tests — the dominant lethal, heritable translocation, and morphological specific locus tests — from which most germ cell mutagenicity data historically have been obtained. Of the 21 anticancer drugs reviewed, 16 have been tested in one or more of these three tests; with all 16 tested in the most common germ cell test, the male dominant lethal test, and 9 of the 16 also tested in the female dominant lethal test. The patterns of germ cell stage specificity for most of the anticancer drugs are similar, and generally resemble the patterns seen with other types of chemicals; however, some of the patterns are unique. For example, 2 of the 8 chemicals shown to induce dominant lethal mutations in female oocytes, do not induce dominant lethal mutations in male germ cells (adriamycin and platinol). Ten of the 16 chemicals tested in the dominant lethal test were positive in post-meiotic stages (spermatids through mature sperm), and seven also induced reciprocal translocations and/or specific locus mutations in post-meiotic stages. This propensity to induce mutations in post-meiotc stages has been observed with most mutagens. However, 5 of the anticancer drugs also induced dominant lethal mutations in spermatocytes (meiotic prophase cells) and one of them, 6-mercaptopurine, uniquely induced dominant lethal mutations exclusively in preleptotene spermatocytes. Finally, three of the anticancer drugs (melphalan, mitomycin C, procarbazine) are members of a very select group of chemicals shown to induce specific locus mutations in spermatogonial stem cells of mice. The implications for human risk are discussed.

Dominant lethal and heritable translocation tests with chlorambucil and melphalan in male mice

Chemicals used in the treatment of cancer include several that are potent mutagens in a range of in vitro and in vivo assays. For some, genetic effects have also been demonstrated in humans, detected as chromosomal aberrations in peripheral lymphocytes. Because (1) many of these agents are confirmed mutagens, (2) humans are exposed to them in relatively high doses, and (3) an increasing number of early cancer victims are surviving to reproductive age, it is important that information be available on the genetic and reproductive hazards associated with exposure to these agents. Chlorambucil and melphalan are structurally related chemicals that are included in our efforts to identify and assess such hazards among cancer chemotherapy agents. To date, both have been reported to induced specific locus mutations in germ cells of male mice (Russell et al., 1989; Russell et al., 1992b) and melphalan is one of very few chemicals shown to induced such mutations in spermatogonial stem cells. More recently, both chemicals were found to have strong reproductive effects in female mice (Bishop and Generoso, 1995, in preparation). In the present studies, these chemicals were tested for the induction of dominant lethal mutations and heritable translocations in male mice. Both chemicals were found to have reproductive effects attributable to cytotoxicity in specific male germ cell stages and to induce dominant lethal mutations and heritable translocations in postmeiotic germ cells, particularly in mid to early stage spermatids. Thus, relatively extensive data are now available for assessing the genetic and reproductive hazards that may result from therapeutic exposures to these chemicals.

The rodent dominant lethal assay: Regulatory questions concerning appraisal of an industrial chemical as exemplified by 1,3-butadiene

Ashby has recently described a clear diagrammatic model for the presentation and assessment of test data from dominant lethal tests. From a regulatory perspective, this approach has the merit of providing data in a standard format. Having recently evaluated the dominant lethal test data for butadiene, we have identified several areas for consideration and discussion.

The performance of short-term tests in identifying potential germ cell mutagens: a qualitative and quantitative analysis

A retrospective analysis was undertaken to assess the performance of selected short-term tests in the discrimination of mammalian germ cell mutagens and nonmutagens using data derived from the U.S. Environmental Protection Agency/International Agency for Research on Cancer Genetic Activity Profile (EPA/IARC GAP) and EPA GENE-TOX databases. The short-term tests selected were gene mutation in Salmonella ( S. typhimurium), cultured mammalian cell gene mutation and chromosomal aberrations, and mammalian bone marrow cytogenetics (micronucleus and chromosomal aberrations). These are the first level tests used in the EPA mutagenicity testing guidelines. The results of this analysis showed good sensitivity of short-term in vitro tests for mammalian cell gene mutation (96%) or chromosomal aberrations (92%) in identifying germ cell mutagens, while the sensitivity of tests for gene mutation in S. typhimurium was lower (79%). Bone marrow micronucleus or chromosomal aberration assays in vivo each displayed a sensitivity of 96%. Thus, both the in vitro and in vivo tests may be used effectively to screen chemicals for potential germ cell mutagenicity. In contrast, the in vitro tests mentioned above performed poorly in discriminating putative germ cell nonmutagens, giving results for specificity at or below what is expected due to chance alone (50–11%). The bone marrow assays were more efficient in this regard, the micronucleus test yielding a specificity of 63% and the chromosomal aberrations assay 64%. The mouse bone marrow micronucleus test also performed well on a quantitative basis, responding at or below the lowest effective doses tested in the mouse dominant lethal assay. Regression analysis of the mean lowest effective doses of chemicals evaluated in vivo showed approximately 1:1 linear correlations for mouse germ cell assays (heritable translocation vs dominant lethal or specific locus tests) as well as for mouse bone marrow assays (micronucleus vs chromosomal aberration). The results suggest the value of the bone marrow micronucleus test as an assay for potential germ cell mutagenicity and the dominant lethal test as a relatively inexpensive choice for confirmation of germ cell damage. The sensitivity of the in vitro assays investigated and the discriminatory capability of the in vivo bone marrow assay affirmed the utility of these tests within the framework of the EPA mutagenicity testing guidelines.

Summary report of the working group on mammalian germ cell tests

The two tests considered by the Working Group were the mammalian germ cell cytogenetic assay and the rodent dominant lethal test. It was agreed that both tests were mainly used for identification of germ cell hazards, however, that the commonly applied protocol of the dominant lethal assay often supplied information for hazard characterization such as sensitivity of particular developmental stages of male germ cells. No particular species or strains were indicated. Concurrent solvent controls were regarded as indispensable for both tests. In the discussion of the mammalian germ cell cytogenetic assay, harmonization was obtained to a large extent with the cytogenetic bone marrow assay regarding the number of animals (5), the number of cells analyzed per animal (200), the highest exposure dose (MTD) and sampling times (twice within 24 and 48 h after dosing). However, it was pointed out that only the single acute exposure was adequate for the mammalian germ cell cytogenetic assay. Furthermore, it was stated that only structural chromosome aberrations could be analyzed and that it was not informative to score polyploidies or aneuploidies. In the discussion of the rodent dominant lethal test, it was stated that the assay was generally performed with treated males, however, increasing concern about female specific effects required that a protocol for female dominant lethal testing should be developed and validated. Acute and subacute treatment schedules were considered equally acceptable. It was regarded as highly important that the entire male germ cell development from meiosis to mature sperm was covered in the test protocol either by the appropriate mating schedules after single dosing or by subchronic dosing during the respective period. Postimplantation loss, preimplantation loss and fertility rate were the main parameters to be assessed in the rodent dominant lethal tests. It was agreed that the size of the experiment depended on the spontaneous frequency of dead implants, the mating scheme and the statistical design of the experiment.

Genotoxicity assessment of the antifungal antibiotic aureofungin in Salmonella typhimurium and Swiss albino mice

The widely used agricultural antifungal agent aureofungin (ARF) was subjected to genotoxicity assessment using the Ames Salmonella assay as well as the in vivo micronucleus test and dominant lethal test in Swiss mice. In the Ames Salmonella spot test, ARF slightly elevated the number of histidine revertants after metabolic activation over a wide dose range (1–1000 μg/plate) in TA102 but not in TA97a, TA98 or TA100. In the preincubation plate incorporation assay with TA102, ARF increased the number of revertants in a dose-dependent manner only after metabolic activation. ARF failed to significantly elevate the frequency of micronucleated polychromatic erythrocytes (PE) in the bone marrow of Swiss mice. It elevated the frequency of dominant lethal mutations in the 7th and 8th weeks at 30 mg/kg body weight, a concentration much higher than the actual concentration used in the field. We conclude that ARF is non-mutagenic in somatic cells in vivo at doses used in the present study, probably mutagenic in stem-cell spermatogonia and may be classified as an equivocal promutagen, possibly acting as a cross-linker.

Cytogenetic and germ cell effects of phosphine inhalation by rodents: II. Subacute exposures to rats and mice.

Phosphine (PH3) is a highly toxic grain fumigant to which there is significant human workplace exposure. To determine the in vivo cytogenetic effects of inhalation of PH3, male F344/N rats and B6C3F1 mice were exposed to target concentrations of 0, 1.25, 2.5, or 5 ppm PH3 for 6 hr/day for 9 days over an 11-day period. Approximately 20 hr after the termination of exposures, blood was removed from the mice and rats by cardiac puncture and the lymphocytes cultured for analyses of sister chromatid exchanges and chromosome aberrations in rats and mice, and micronuclei (MN) in cytochalasin B-induced binucleated lymphocytes from mice. In addition, bone marrow (rats) and peripheral blood (mice) smears were made for the analysis of MN in polychromatic and normochromatic erythrocytes. No significant increase in any of the cytogenetic endpoints was found at any of the concentrations examined. These results indicate that concentrations of PH3 up to 5 ppm are not genotoxic to rodents when administered by inhalation for 9 days during an 11-day period as measured by several cytogenetic assays. To evaluate the effects of PH3 on male germ cells, a dominant lethal test was conducted in male mice exposed to 5 ppm PH3 for 10 days over a 12-day period and mated to groups of untreated females (2 females/male) on each of 6 consecutive 4-day mating intervals. None of the 6 groups of females exhibited a significant increase in percent resorptions. These results indicate that exposure to 5 ppm PH3 by inhalation does not induce dominant lethality in male mouse germ cells at steps in spermatogenesis ranging from late differentiating spermatogonia/early primary spermatocytes through mature sperm.

Dominant-lethal mutations and micronucleus induction in male BALB/c, BDF1 and H mice by tobacco smoke

The mutagenicity of whole tobacco smoke (TS) was examined in male BALB/c, BDF1 and H mice using the dominant lethal and micronucleus tests in bone marrow polychromatic erythrocytes. Male mice (about 80 days old) from the three strains were treated with TS on 5 days/week for 8 weeks. The animals were divided into three groups for every strain: control and two experimental groups. Two doses - low (1h treatment/day) and high (2 h treatment/day) - were used. In the first case 8 exposures of 7.5 min each with 1-min intervals were given and to realize the high dose this was repeated 4 h later. It was found that TS induces significant dominant-lethal mutations in both experimental groups of BALB/c, BDF1 and H mice ( p < 0.001), but some strain differences existed. The data obtained suggest that in BALB/c and BDF1 mice TS induces dominant-lethal mutations mainly in spermatocytes, spermatogonia and gonial stem cells, while in H mice only spermatids and spermatocytes are affected. The bone marrow from each strain and each dose group was investigated for the presence of micronucleated polychromatic erythrocytes (PCE) at 5, 19, 38, 54, and 63 days after beginning of the treatment with TS, using the same exposure regimen. Exposure of male mice to TS caused an up to 2–3-fold increase in the number of PCE in the bone marrow of BALB/c and BDF1 mice in both dose groups. In H mice this effect is observed only on days 19 and 38 of sampling. No cumulative or dose-dependent effects were detected. Increased formation of micronuclei within PCE of femoral bone marrow after passive smoking was regarded as being due to the effect of TS.

Effect modification by inorganic lead in the dominant lethal assay

Experimental evidence suggests that inorganic lead may modify mutagenic events. We examined the modifying effect of lead on mutagenic events in late spermatogenesis in the dominant lethal assay. Twelve NMRI male mice were given lead chloride in the drinking water and 12 male mice received tap water without lead chloride. Cyclophosphamide (120 mg/kg b.w., i.p.) was given to six males in the lead treatment group and six males in the tap water group 1 week before mating. This resulted in four treatment groups: control, lead, cyclophosphamide, and lead plus cyclophosphamide. Cyclophosphamide given to the males (with or without lead treatment) reduced the numbers of live implants in mated females. The most prominent effect of cyclophosphamide was an increase of resorbed implants. Females mated to lead exposed male mice showed a nonsignificantly lower frequency of resorptions compared to controls. The results give no support to the hypothesis that inorganic lead may influence the mutagenicity of cyclophosphamide in the dominant lethal test.

Trichlormethine hydrochloride and correlation of its mutagenic and toxic effects on male germ cells in mice

The genetic effect of the cytostatic trichlormethine hydrochloride (TS-160 Spofa) was assessed after a 1-week administration using the dominant lethal mutation test (DLM) and the sperm abnormality test. The dosage was 0.5 mg/kg for 7 consecutive days, an equivalent of the human therapeutic dosage. Simultaneously, the cytostatic's direct toxic effect on male sex organs was assessed. TS-160 carries a genetic risk for the postmeiotic stages of spermatogenesis (DLM) and is responsible for interference in the morphology of sperm heads through its action on spermatocytes. The toxic effects of TS-160 were found to influence the body weight of mice (days 4–25 after administration), to reduce the relative weight of the testes (days 18–25 after administration), to damage spermatogenesis in the seminiferous tubules (spermatids), to be responsible for an appearance of multinucleate cells in the epididymides, and for an increased rate of abnormality of the heads of fully mature spermatozoa. Our findings stress the need to separate the cytotoxic effects from genetic effects so as to avoid false positives, especially in the test for head abnormalities, and also in the assessment of the fertility of male animals or fertilization of females mated with treated males.