Masaaki Moriya

Further mutagenicity studies on pesticides in bacterial reversion assay systems

A total of 228 pesticides (88 insecticides, 60 fungicides, 62 herbicides, 12 plant-growth regulators, 3 metabolites and 3 other compounds) was tested for mutagenicity in bacterial reversion-assay systems with 5 strains (TA100, TA98, TA1535, TA1537 and TA1538) of Salmonella typhimurium and a strain (WP2 hcr) of Escherichia coli. 50 pesticides (25 insecticides, 20 fungicides, 3 herbicides, 1 plant-growth regulator and 1 other compound) were found to be mutagenic. 5 of them required metabolic activation (S9 mix) for their activities. Among various chemical groups, organic phosphates, halogenated alkanes and dithiocarbamates showed higher ratios of mutagens. Although 22 of the pesticides tested have been reported to be carcinogenic, 7 of them, i.e., captain, DBCP, EDB, EDC, ETU, HEH and nitrofen, were detected as mutagens in the present assay. Most of the other 15 non-mutagenic carcinogens were organochlorine pesticides such as alpha-BHC, chlorobenzilate, p,p-DDT, dieldrin and quintozene.

Mutagenicity screening of pesticides in the microbial system

A survey on the mutation induction capacity was made in the microbial system on 166 pesticides including 57 fungicides, 63 herbicides and 46 insecticides. The screening methods consisted of the rec-assay procedure, a sensitivity test utilizing H17 Rec+ and M45 Rec- strains of Bacillus subtilis, as well as the reversion assays on plates utilizing auxotrophic strains of Escherichia coli (WP2) and Salmonella typhimurium (Ames series). Chemicals inducing reversions were detected only among those showing positive effects in the rec-assay but not among negative samples. In addition to Captafol, Captan, Dexon and NBT of which mutagenicities have been previously reported, Dichlorvos, Folpet, 2-hydrazinoethanol (HEH), 5-nitro-1-naphthonitrile (NNN) and Vamidothion were found to be mutagens in our systems.

Mutagenicity screening of feed additives in the microbial system

18 feed additives were tested for DNA-modifying effects by the repair test named rec-assay with Bacillus subtillis H17 (rec+) and M45 (rec-), and for mutagenicity with Escherichia coli WP2 hcr and 5 Salmonella typhimurium tester strains with the use of a top-agar overlay method. Carbadox, furazolidone, panazon and zoalene were positive in both assays. The former 3 were mutagenic for TA100, TA98 and WP2 hcr, while zoalene was mutagenic for all strains. These 4 compounds did not require a metabolic activation for their mutagenic activities. Nicarbazin was weakly mutagenic for TA1538 and TA98 with and without S9 mix. Amprolium and caprylohydroxamic acid also showed very weak mutagenicities only for TA100 with S9 mix and for WP2 hcr with and without S9 mix, resp. The mutagenic activities of carbadox, furazolidone and panazon for TA100 were reduced only by the addition of S9 mix, but not by S9 fraction or blood, whereas that of zoalene was decreased by any of the 3 factors.

Mutagenicity of Pesticides

Microbial mutagenicity screening studies employing Ames tests were done on 228 pesticides. As a result, 50 compounds showed mutagenicity and five of them required metabolic activation for their activities. Among the various compound groups, organic phosphates, halogenated alkanes and dithiocarbamates contained mutagens at a higher ratio. Mutagenicity of these pesticides was less potent than that of other mutagens. Cytogenetic studies on the pesticides which were positive in microbial assays revealed a good correlation between the ability to induce sister chromatid exchanges or chromosomal aberrations and the mutagenic potency in bacteria. Dominant lethal studies on 1, 2-dibromo-3-chloropropane (DBCP) and ethylene dibromide (EDB) disclosed that DBCP gave positive results in rats but not in mice, although EDB was negative in both species. DBCP was also positive in a sex-linked recessive lethal test using Drosophila melanogaster. Occupational exposure to pesticides, especially gaseous ones like DBCP, in industry or in agriculture seems hazardous from the genetic toxicological point of view.

Inhibitors for the mutagenicities of colon carcinogens, 1,2-dimethylhydrazine and azoxymethane, in the host-mediated assay.

Inhibitory effects of several chemicals on the mutagenicities of 1,2-dimethylhydrazine (DMH) and azoxymethane (AOM) for Salmonella typhimurium G46 in the host-mediated assay were investigated. They were carbon disulfide (CS2), tetraethylthiuram disulfide (disulfiram, DSF), sodium diethyldithiocarbamate (SDDC), ethylene-bis(dithiocarbamato) manganese (Maneb), pyrazole (PZ), aminoacetonitrile hydrogen sulfate (AAN), and sodium selenite (SE). All the compounds, except for SE, inhibited the mutagenicities of DMH and AOM.

Detection of mutagenicity of procarbazine by the host-mediated assay with polychlorinated biphenyl (aroclor 1254) as enzyme inducer

Procarbazine [N-isoppropyl-alpha-(2-methylhydrazino)-p-toluamide] was tested for mutagenicity with Salmonella typhimurium G46 in the host-mediated assay by using male BALB/c mice pretreated orally with polychlorinated biphenyl (PCB, Aroclor 1254). Procarbazine was weakly mutagenic without PCB pretreatment, but the pretreatment greatly enhanced the mutagenicity of this compound. The administration of 500 mg PCB/kg 1 day before procarbazine dosage was suitable for the detection of the mutagenicity. Among PCB, 3-methylcholanthrene (3-MC) and phenobarbital sodium (PB), the former 2 inducers showed much stronger enhancing effects than PB. The pretreatment with 3-MC in combination with PB did not cause further enhancement compared with 3-MC alone.

Carcinogenicity of N-nitroso-ethylenethiourea in female mice

The nitrosation product of ethylenethiourea (ETU), N-nitroso-ETU, was tested for tumorigenicity in female ICR mice. Ten weekly oral administrations of this compound (0.66-2.64 mg/dose) caused apparently dose-dependent increases in the number of mice with pulmonary and lymphocytic neoplasms.

Induction of sex-linked recessive lethal mutations in Drosophila melanogaster males by gaseous 1,2-dibromo-3-chloropropane (DBCP)

Acute exposure to gaseous 1,2-dibromo-3-chloropropane (DBCP) caused sex-linked recessive lethal mutations in Drosophila melanogaster males. Brood pattern analysis showed that spermatids and spermatocytes were more sensitive to the lethal-mutation induction by DBCP than were spermatogonia. The mutation induction was not observed in the stage of spermatozoa.