William M. Snellings

A two-year inhalation study of the carcinogenic potential of ethylene oxide in fischer 344 rats☆

Fischer 344 rats were exposed to 100, 33, 10, or 0 ppm of ethylene oxide vapor (EtO) by inhalation for 6 hr per day, 5 days per week, for approximately 2 years. Inhalation of EtO resulted in a significant depression of body weight gain in the 100- and 33-ppm exposure groups and a significant increase in mortality in the 100-ppm group. Through 18 months of exposure to EtO, no statistically significant increases in tumor incidence were observed. After 18 months, the incidence of primary brain tumors was increased for both sexes. Statistical evaluation indicated a treatment-related response, particularly for the male rats, in the 100- and 33-ppm exposure groups. After 24 months of exposure, histologic findings confirmed hematologic evidence that exposure to EtO resulted in an increased prevalence of mononuclear cell leukemia, which is a neoplasm that is common in aged Fischer 344 rats. This increase was dose related and increased for each of the three exposure concentrations. The percentage of female rats with multiple neoplasms was also greater in all three exposure groups than in controls. Furthermore, in both the 100- and 33-ppm exposure groups, the percentage of female rats with at least one malignant neoplasm was increased. An increased frequency of peritoneal mesothelioma was treatment related in the male rats exposed to 100 or 33 ppm of EtO. This study has shown biologically significant adverse effects at all concentrations tested. The incidences of mononuclear cell leukemia, peritoneal mesothelioma, and primary brain tumors in the air-control rats were similar to those reported in the literature. The possible contribution of a sialodacryoadenitis viral outbreak (which occurred during the 15th exposure month) to the EtO exposure-related tumors is unknown, though unlikely.

Ethylene glycol monobutyl ether: Acute, 9-day, and 90-day vapor inhalation studies in Fischer 344 rats

The acute 4-hr LC50 (with 95% confidence limits) for Fischer 344 rats was determined to be 486 (339 to 696) ppm of ethylene glycol monobutyl ether (EGBE) for males and 450 (315 to 645) ppm for females. Notable observations included loss of coordination, red stained urine, and enlarged discolored kidneys at 867 and 523 ppm. In a subsequent study, rats were exposed for 9 days (6 hr/day) to EGBE concentrations of 245, 86, 20, or 0 (control) ppm. There were significant depressions of red blood cell (RBC) count (approximately 20% below control values), hemoglobin (Hgb), and mean corpuscular hemoglobin (MCH) concentration and increases in nucleated erythrocytes, reticulocytes, and lymphocytes in males and females of the 245 ppm group. Decreased body weight gains and increased liver weights were also found. A 14-day postexposure recovery showed substantial reversal of the affected blood parameters. Similar, but less marked, hematologic effects were observed in rats exposed to 86 ppm of EGBE, while rats of the 20 ppm group were indistinguishable from controls. In a 90-day study, rats were exposed to EGBE concentrations of 77, 25, 5, or 0 ppm for 13 weeks (6 hr/day, 5 days/week). Slight, but statistically significant, decreases in RBC (13% below control) and Hgb, accompanied by an increase in MCH (11% above control) were observed in the 77 ppm-exposed females after 6 weeks. At the conclusion of the 90-day exposure regimen, the hematologic effects seen in the females had lessened (RBC was 7% below control) or returned to control value ranges. Furthermore, no treatment-related differences were found in body weight, organ weights, urine or serum chemistries, gross lesions, or microscopic lesions in males or females. There were no significant biological effects in rats exposed subchronically to 25 or 5 ppm. The subtle hematologic findings of these studies confirm the known RBC perturbations of EGBE.

Acute inhalation studies with methyl isocyanate vapor: I. Methodology and LC50 determinations in guinea pigs, rats, and mice

Groups of male and female Fischer 344 rats, B6C3F1 mice, and Hartley guinea pigs were exposed once for 6 hr to mean concentrations of 10.5, 5.4, 2.4, 1.0, or 0 (control) ppm of methyl isocyanate (MIC) vapor. Rats and mice were also exposed to 20.4 ppm of MIC. No deaths occurred in animals exposed to 2.4 or 1.0 ppm. The majority of deaths for the 20.4- and 10.5-ppm groups occurred during postexposure Days 1 through 3, while at 5.4 ppm deaths were observed throughout the 14-day postexposure period. The 6-hr LC50 values (with 95% confidence limits) were 6.1 (4.6 to 8.2) ppm for rats, 12.2 (8.4 to 17.5) ppm for mice, and 5.4 (4.4 to 6.7) ppm for guinea pigs. Notable clinical observations during and immediately following MIC exposure were lacrimation, perinasal/perioral wetness, respiratory difficulty (e.g., mouth breathing), decreased activity, ataxia, and hypothermia. The frequency of clinical signs decreased during the second postexposure week. Body weight losses were common in all species following MIC exposures of 2.4 ppm or greater. At 1.0 ppm, only female mice had body weight depression. Recovery of body weight loss was observed in the 5.4- (guinea pigs only), 2.4- and 1.0-ppm concentration groups. The lungs of all animals that died were discolored. Following microscopic examination of the respiratory tract, deaths were attributed to pulmonary edema and congestion. In a separate study, Fischer 344 rats and Hartley guinea pigs were exposed once for 4 hr to mean concentrations of 36.1, 25.6, 15.2, or 5.2 ppm of MIC vapor. In general, the results were similar to those of the single 6-hr exposure study.

Teratology study in fischer 344 rats exposed to ethylene oxide by inhalation

Abstract Pregnant Fischer 344 rats were exposed 6 hr/day to either 100, 33, or 10 ppm of ethylene oxide vapor on Days 6 through 15 of the gestation period. Two separate control groups were maintained under the same conditions but were exposed to room air only. Two other groups of rats were given a known rodent teratogen, aspirin, by gavage; one group was given 500 mg/kg body weight on Day 9 of gestation, and the other was given 625 mg/kg body weight on Day 10 of gestation. No treatment-related effects of ethylene oxide were noted for the following evaluations; maternal survival, litter size, number of implantation and resorption sites, and number of preimplantation losses. Exposure to 100 ppm of ethylene oxide resulted in a significant depression of body weight in the fetuses, but did not result in any abnormal effects of embryonic or fetal lethality, or teratologic effects of the soft tissues or skeleton. Consequently, ethylene oxide was not considered a teratogen by inhalation in the Fischer 344 rat at an exposure concentration of 100 ppm.

Effects on reproduction in Fischer 344 rats exposed to ethylene oxide by inhalation for one generation

Abstract After 12 weeks of exposing male and female Fischer 344 rats (6 hr/day, 5 days/week) to either 100, 33, or 10 ppm of ethylene oxide vapor, these animals were mated, and the females were continued on exposure from Day 0 through Day 19 of gestation (6 hr/day, 7 days/week). The major treatment-related adverse effect was significantly fewer pups born per litter in the highest exposure level only. There were fewer implantation sites per pregnant female, and a smaller ratio of the number of fetuses born to the number of implantation sites per pregnant female in the 100-ppm exposure group than in any other group. No statistically significant differences of male or female fertility indices were noted between the ethylene oxide exposure and air-control groups. There were significantly more animals with a gestation period longer than 22 days in the 100-ppm exposure group when compared to either air-control group. No significant differences in body weight gain or any other signs of toxic effects from exposure were observed in the F0 generation. Moreover, there was no treatment-related adverse effect on survival or growth rate of the Fla generation during the lactation period, even when the dams were exposed to ethylene oxide during this period.

A subchronic inhalation study on the toxicologic potential of ethylene oxide in B6C3F1 mice.

Four groups of B6C3F1 mice, each containing 30 per sex, were exposed to ethylene oxide (EtO) vapor at target concentrations of 250, 100, 50, 10, or 0 ppm which are equivalent to 450, 180, 90, 18, and 0 mg/m3, respectively. The actual mean chamber concentrations were 236, 104, 48, 10, and 0 ppm, respectively. After an exposure regimen of 6 hr per day, 5 days per week, for 10 weeks (males), or 11 weeks (females), urine and blood samples were taken for clinical pathologic evaluation and selected tissues were weighed and examined by light microscopy. Statistically significant findings that could be indicative of a toxic response were observed in the 250-ppm exposure group. These included minimal changes in certain erythroid parameters, increased liver weight, decreased testicular weight, and decreased spleen weight which was noted also in the 100-ppm group. However, there were no microscopic findings to support or explain any of these apparently treatment-related effects. Results of a neuromuscular screening test indicated that certain reflex responses and locomotor activities were altered in the EtO-exposed animals. A dose-related trend of response in the 250-, 100-, and 50-ppm exposure groups was noted in the evaluation of locomotor function; however, because of the small sample size, it was not possible to determine a threshold concentration for this effect. There were no accompanying histopathologic alterations in muscle and central or peripheral nervous tissue.

Subchronic Toxicity Study of Dicyclopentadiene Vapor in Rats

Fischer 344 rats were exposed by inhalation to 0, 1, 5 or 50 ppm dicyclopentadiene (DCPD) vapor 6 hr/day, 5 days/week for 13 weeks, followed by a 13-week recovery period. Animals were euthanized following completion of exposure at 2, 6, or 13 weeks and at postexposure weeks 4 or 13. No mortality, overt signs, body weight changes, hematologic or clinical chemistry values were related to DCPD exposure. In the high-exposure male rats, relative liver weights were significantly increased but with no accompanying histopathologic changes. Exposure to DCPD produced adverse kidney effects in male, but not female, rats as evidenced by the excretion of epithelial cells in the urine. Histologic changes were localized to the proximal tubules of the kidney and included increased accumulation of protein droplets, regenerative epithelium, and the presence of intraluminal proteinaceous material. In addition, several alterations in renal function were observed. Urinary Na+ excretion rates were decreased and urinary K+ excretion rates were increased throughout the exposure period; however, glucose was not present in the urine, and creatinine clearance was normal. The ability of the kidney to concentrate urine was also impaired. After the recovery period, many of the treatment-related kidney effects were not observed, including the presence of hyaline droplets in the proximal tubules and epithelial cells in the urine. These findings indicate an overall low degree of systemic toxicity following sub-chronic inhalation exposure of dicyclopentadiene at exposure levels up to 50 ppm. The only effect that was observed was a male rat-specific nephropathy that is characteristic of the hyaline droplet nephropathy produced by a diverse group of compounds.

2,4-Pentanedione: 9-Day and 14-week vapor inhalation studies in Fischer-344 rats

Fischer-344 rats, in groups of 10 males and 10 females, were exposed for 9 days (6 hr/day) to 2,4-pentanedione (2,4-PD) vapor at mean concentrations of 805, 418, 197, and 0 (control) ppm. No deaths occurred, and the only adverse signs were of sensory irritation (partial closure of eyelids, periocular and perioral wetness) at 805 ppm. Also at 805 ppm were decreased body and organ weights, lymphocytosis, and moderate inflammation of the nasal mucosa. At 418 ppm there was a decrease in body weight gain and mild inflammation of the nasal mucosa. Apart from minimal nasal mucosal inflammation, there were no effects at 197 ppm. In the subchronic (14-week) study, rats were exposed (6 hr/day; 5 days/week) to 650, 307, 101, and 0 (control) ppm of 2,4-PD vapor, using groups containing 20 males and 20 females, with half being sacrificed at the end of the exposure period and the remainder kept for a 4-week postexposure recovery period. An additional 10 males were added to the 650 and 0 ppm groups for glutaraldehyde perfusion and subsequent electron microscopic examination of sciatic nerves. At 650 ppm, all females and 10 of 30 males died between the second and sixth weeks of exposure. These animals had acute degenerative changes in the deep cerebellar nuclei, vestibular nuclei and corpora striata, and acute lymphoid degeneration in the thymus. Seven of 15 male survivors of the 650 ppm group (combined 14-week and recovery sacrifices) had gliosis and malacia in the same brain regions, minimal squamous metaplasia in the nasal mucosa, decreased body and organ weights, lymphocytosis, and minor alterations in serum and urine chemistries. No ultrastructural evidence of peripheral neuropathy was observed. Except for central neuropathy, many of the adverse effects at 650 ppm were less marked in the 4-week recovery animals. No deaths occurred at 307 ppm, but females had slightly decreased body weight gains, and in both sexes there were minor alterations in hematology, serum chemistry, and urinalysis parameters, which were not present in the 4-week recovery animals. Rats exposed to 101 ppm showed no differences from the control rats. Subchronic exposure to 650 ppm of 2,4-PD vapor causes serious adverse biological effects. Under these study conditions, the minimum-effects concentration was 307 ppm, and the no-adverse effects concentration was 101 ppm.

Monochlorodiisobutylene vapor: Acute and 9-day inhalation studies in Fischer-344 rats and B6C3F1 mice☆

Acute and 9-day repeated exposures to monochlorodiisobutylene (CDIB) were conducted in male and female Fischer-344 rats and B6C3F1 mice. The 4-hr LC50 values for these animals ranged between 1400 and 2100 ppm. Animals in the 9-day study were exposed at a mean concentration of 478, 97, or 25 ppm of CDIB for 6 hr per day. Treatment-related effects differed between species in this study. Body weight change was decreased in rats. Morphologic changes in the kidneys with accompanying polyuria and hematuria/hemoglobinuria were observed in male rats. The only effect observed at 25 ppm was a low incidence of hematuria/hemoglobinuria in male rats. Mice appeared unaffected by exposure to CDIB at levels as high as 478 ppm.