Evaluation of the mutagenicity of acetochlor to male rat germ cells.
Male rat dominant lethal (DL) assays conducted on the herbicide acetochlor are described. Single dose studies conducted at the maximum tolerated dose (MTD, < or = 1000 mg/kg) produced no effects on any of the DL assay parameters at any of the ten weekly sampling periods. It is concluded that acetochlor is non-mutagenic to rat germ cells. Due to initial limited knowledge of the MTD of acetochlor it was also evaluated in the DL assay at a dose level of 2000 mg/kg. At this high dose level severe bodyweight loss and some deaths occurred among the treated animals. In addition, reduced implantations and reduced pregnancy rates were observed at the third sampling period (18-25 days post dosing) in the absence of an increase in early post-implantation deaths. These results indicated that the use of supra-MTD doses of acetochlor had reduced the fertility of the treated males leading to the production of a pseudo-DL assay response, as alerted to and defined by Ehling. Although several such pseudo-DL assay responses have been described, none have been explained mechanistically. It was therefore decided to pursue the effects seen in the DL assay when using supra-MTD doses of acetochlor. Ova analysis of female rats mated with male rats exposed to 2000 mg/kg acetochlor revealed unfertilized ova at the critical third sampling time. Normal fertilization of ova was observed at the first and fifth sampling period and, for a dose of 200 mg/kg acetochlor, at the third sampling period. The magnitude and temporal nature of these effects confirmed the induction of a pseudo-DL assay response, and studies were then undertaken to probe its genesis. Rats treated with 2000 mg/kg acetochlor had normal testicular and epididymal pathology and normal sperm numbers and sperm motility at the critical third sampling period. Despite a small reduction in testicular and epididymal glutathione levels 12 h after exposure to 2000 mg/kg acetochlor, testicular LDH and LDH-X enzyme levels were unaffected. Further, no reduction in the level of free sulphydryl groups (-SH) were observed in epididymal caput sperm heads isolated 0.5, 7 or 14 days after treatment of male rats with 2000 mg/kg acetochlor. The only sperm parameter affected by treatment with 2000 mg/kg acetochlor was an increase in epididymal cauda sperm with head abnormalities. The non-specific nature of this effect was considered inadequate to explain fully the high dose fertility effects seen in the DL assays, which therefore remain unexplained. The present data establish that acetochlor is non-mutagenic to rat germ cells. They also confirm the importance of segregating mutagenic and fertility effects in the DL assay, and emphasize the need for appropriate dose-setting studies prior to the conduct of rodent genetic toxicity assays.
Ashby
J
J
Zeneca Central Toxicology Laboratory, Alderley Park, Cheshire, UK.
Tinwell
H
H
Lefevre
P A
PA
Williams
J
J
Kier
L
L
Adler
I D
ID
Clapp
M J
MJ
D000818:Animals; D001835:Body Weight / Q000187:drug effects; D005260:Female; D005799:Genes, Dominant / Q000187:drug effects* / Q000235:genetics; D005804:Genes, Lethal / Q000187:drug effects* / Q000235:genetics; D005854:Germ Cells / Q000187:drug effects* / Q000473:pathology; D005978:Glutathione / Q000032:analysis; D006540:Herbicides / Q000633:toxicity*; D007091:Image Processing, Computer-Assisted; D007770:L-Lactate Dehydrogenase / Q000187:drug effects / Q000378:metabolism; D008297:Male; D016296:Mutagenesis / Q000187:drug effects*; D009929:Organ Size / Q000187:drug effects; D010063:Ovum / Q000187:drug effects; D011247:Pregnancy; D051381:Rats; D013077:Sperm Head / Q000187:drug effects / Q000473:pathology; D013737:Testis / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D014052:Toluidines / Q000633:toxicity*
D006540:Herbicides; D014052:Toluidines; C043377:acetochlor; D007770:L-Lactate Dehydrogenase; D005978:Glutathione