Developmental toxicity study of inhaled acrylic acid in New Zealand White rabbits.

In range-finding and definitive developmental toxicity studies, timed pregnant New Zealand White rabbits were exposed to acrylic acid (CAS No. 79-10-7) vapour for 13 consecutive days during pregnancy. In the range-finding study, eight pregnant does/group were exposed to 30, 60, 125 or 250 ppm acrylic acid vapour on gestation days (gd) 10-22 of pregnancy. Monitors of toxicity included body weight measurements, daily food consumption measurements and clinical observations. Three of the eight does/group were killed on the day following the last exposure (gd 23), and the remaining does were killed and autopsied on gd 29. At autopsy, special attention was given to gross observation of maternal nasal turbinates, and nasal turbinates from all does were evaluated histologically. No evaluation of foetuses was performed in the range-finding study. In the definitive study, 16 does/group were exposed to concentrations of 25, 75 or 225 ppm acrylic acid vapour from gd 6 to 18, the major period of organogenesis. Monitors of maternal toxicity included clinical observations and measurements of body weight and daily food consumption measurements. Does were killed and autopsied on gd 29. Maternal liver and kidney weights were measured and external, visceral and skeletal evaluations of foetuses were conducted. Maternal nasal turbinates were not evaluated histologically in the definitive study. Effects in does from both studies included consistent concentration-related reductions in food consumption and body weight gains throughout the exposure period at concentrations of acrylic acid vapour above 60 ppm. Characteristic clinical signs of sensory irritation, including perinasal and perioral wetness and severe nasal congestion, were noted in does from both studies at or above vapour concentrations of 75 ppm. Gross observation of nasal turbinates immediately following exposures in the range-finding study indicated colour changes in the nasal turbinates of does in the 60 and 250 ppm groups. Colour changes in the nasal turbinates were noted in one doe from the 250 ppm exposure group killed on gd 29. Pertinent autopsy findings in the does from the definitive study included ulceration of the nasal turbinates of a single doe in the 225 ppm group. Histological evaluation of turbinates from does killed the day following exposures in the range-finding study revealed lesions in the nasal epithelium in all acrylic acid-exposed groups. The severity of the lesions was concentration related. Microscopic evaluation of turbinates from does killed on gd 29 showed the presence of nasal lesions in the 60, 125 and 250 ppm groups. However, the nasal tissues had recovered considerably during the post-exposure interval. Despite the severe effects on the nasal mucosa of does in both studies, there was no evidence of developmental toxicity including teratogenicity at any exposure concentration used in the definitive study.

Development of pituitary lesions in ND4 Swiss Webster mice when estimating the sensory irritancy of airborne chemicals using ASTM method E981-84.

This study determined the origin of pituitary lesions found in male ND4 Swiss Webster mice following a single head-only exposure to inhaled test materials using ASTM E981-84, standard test method for estimating sensory irritancy of airborne chemicals. Necropsy and histopathology data were evaluated due to the occurrence of unexpected pituitary lesions in sham control and exposure groups. Groups of four mice were restrained in body plethysmographs and exposed for 30 min to increasing dust concentrations of one of three test chemicals to assess the ability to cause sensory irritation. Sham control and test material-exposed mice were sacrificed after a single exposure and subjected to a complete necropsy and microscopic evaluation of the pituitary gland. Control mice remained in the animal room and were not restrained in the plethysmograph. Gross observation at necropsy showed pituitary lesions in one of seven unrestrained control mice (revised to zero of seven after microscopic examination). Seven of seven sham control mice had pituitary lesions, suggesting that the lesions were not related to test material exposure. Each test material-exposed group also had pituitary lesions with high incidence (52/60 for all groups combined), which was not exposure concentration-dependent. Microscopic evaluation of the pituitary glands revealed that darkening of the gland was due to hemorrhage and confirmed that the lesions developed with 100% incidence (19/19) in plethysmograph-housed animals. The rubber neck seal used to restrict animal movement in the plethysmograph appears to have caused an increase in pressure in the blood vessels in the pituitary gland; vessels then ruptured and hemorrhaged. This finding should not adversely affect sensory irritation responses evaluated with this method.

Subchronic dermal toxicity study of triethanolamine in C3H/HeJ mice.

Triethanolamine (TEA) was applied to the skin of male and female C3H mice (15 per sex per dose group) three times weekly for 95 days (37 applications). TEA was administered at concentrations of 0 (acetone vehicle), 10, 33 and 100% (undiluted) in a volume of 50 microliters. The approximate daily doses of TEA were 0.14, 0.46 or 2.0 g/kg per male and 0.16, 0.54 or 2.3 g/kg per female, respectively. The animals were weighted weekly and observed for clinical signs including skin irritation. 10 mice per sex per dose group were designated for clinical chemistry and haematology at terminal killing. Complete autopsies were performed, and the liver, kidneys, brain, heart, spleen, thymus and testes were weighted. Histopathology was performed on tissues from control and high-dose mice and on target organs. Treatment-related effects were limited to a slight epidermal hyperplasia at the site of application at all TEA concentrations. The results indicate that TEA caused a mild local reaction at all concentrations tested, but did not cause systemic toxicity under these conditions.

Hepatic ultrastructural changes induced by the toxin microcystin-LR (MCLR) in mice.

Microcystin-LR (MCLR) is a cyclic heptapeptide produced by the blue-green algae Microcystis aeruginosa. It is highly toxic and causes death in rodents due to hypovolemic shock with associated intrahepatic hemorrhage. The molecular mechanism of toxicity is unknown. In order to provide additional information regarding the toxicity of MCLR, the ultrastructural changes present in livers of mice following the administration of 100 micrograms MCLR/kg intraperitoneally, (i.p.) were examined. Time-dependent changes were observed. Disruption of cell to cell contact occurred with infiltration of erythrocytes 60 min after MCLR treatment. Hepatocyte distortion, mitochondrial aggregation, and the prominent accumulation of large areas of endoplasmic reticulum were observed. No detectable hepatic changes in sinusoidal endothelial cells were present. The results suggest that MCLR preferentially affects hepatocytes, although the observations do not preclude the involvement of hepatic vasculature in the toxicity of MCLR.

Acute vapour inhalation toxicity of acrolein and its influence as a trace contaminant in 2-methoxy-3,4-dihydro-2H-pyran.

1. The LC50 values for acrolein (AC) vapour to Sprague-Dawley rats (combined sexes) were determined to be 26 ppm (1 h) and 8.3 ppm (4 h). Signs of severe irritancy were present, and death was due to lung injury. 2. Exposure of rats to a 2-methoxy-3,4-dihydro-2H-pyran (MDP) saturated vapour atmosphere statistically generated from liquid MDP containing 0.037% AC, caused severe irritancy and death from accumulation of AC vapour. Sparging the impure material with nitrogen gas before atmosphere generation significantly reduced or abolished lethal toxicity. 3. Dynamically generated MDP vapour atmosphere produced transient respiratory and ocular irritancy, but no mortalities. The intrinsic acute vapour inhalation toxicity of MDP is low. 4. The presence of highly volatile toxic impurities in a material may confer a significant acute inhalation toxicity and hazard under conditions of low air movement. Assessment of potential inhalation hazards from liquid mixtures may require investigation by static and dynamic methods for vapour generation.

Dermal oncogenicity studies on two methoxysilanes and two ethoxysilanes in male C3H mice.

The dermal oncogenic potential of beta-(3,4-epoxycyclohexyl)ethyltrimethoxysilane (EEMS), gamma-glycidoxypropyltrimethoxysilane (GPMS), beta-(3,4-epoxycyclohexyl)ethyltriethoxysilane (EEES), and gamma-glycidoxypropyltriethoxysilane (GPES) was assessed by applying 25-microliters aliquots of acetone solutions to the skin of 40 male C3H/HeJ mice. The concentrations applied were 100, 25, 10, and 10% by volume for EEMS, GPMS, EEES, and GPES, respectively. Applications were made thrice weekly until the death of the animals. A negative control group received acetone (solvent) only. No treatment-related skin tumors were observed, nor was there evidence of increased incidence of any internal tumor in the groups that received GPMS, EEES, or GPES. In the group treated with EEMS, four mice were observed with squamous cell carcinomas of the treated skin and two mice had subcutaneous sarcomas outside of the treated area. No skin tumors were observed in the group treated with acetone, but two mice had subcutaneous sarcomas outside of the treated area. The mean survival times were 529, 482, 545, 492, and 502 days for the EEMS, GPMS, EEES, GPES, and acetone control groups, respectively. In no case was the mortality rate significantly different from that of the controls. The results indicate that only EEMS was oncogenic under the conditions of these studies.

Dermal oncogenicity studies on various ethyleneamines in male C3H mice.

The dermal oncogenic potential of diethylenetriamine, high purity and commercial grades (DETA-HP and DE-TA-C), triethylenetetramine (TETA), tetraethylenepentamine (TEPA), and polyamine HPA No. 2 was assessed by applying 25-microliter aliquots of aqueous solutions to the skin of groups of 50 male C3H/HeJ mice. The concentrations applied were 5.0, 5.0, 5.0, 25.0, and 10.0% by volume for DETA-HP, DETA-C, TETA, TEPA, and HPA No. 2, respectively. Applications were made thrice weekly until the death of the animals. A negative control group received deionized water (solvent) only. All animals were individually housed. No treatment-related skin tumors were observed, nor was there evidence of increased incidence of any internal tumor. Twenty TEPA-treated mice had hyperkeratosis and 13 had necrosis of the epidermis, both indicative of skin irritation. Such lesions were absent or occurred very infrequently in the other groups of mice. The mean survival times were 587, 662, 627, 591, 601, and 626 days for the DETA-HP, DETA-C, TETA, TEPA, HPA No. 2, and water control groups, respectively. In no case was the mortality rate significantly different from that of the controls. The results indicate that none of these compounds was oncogenic under the conditions of these studies.

A 14-week vapor inhalation toxicity study of methyl isobutyl ketone.

In a 2-week probe study male and female Fischer-344 rats and B6C3F1 mice were exposed 6 hr/day to 2000, 500, 100, or 0 ppm methyl isobutyl ketone (MIBK). At 2000 ppm there was a slight increase in male rat liver weight (absolute and relative). The only changes observed histologically were increases in regenerative tubular epithelia and hyalin droplets in kidneys of male rats exposed to 2000 or 500 ppm. Exposure levels for a subchronic study were 0, 50, 250, or 1000 ppm methyl isobutyl ketone vapors 6 hr/day, 5 days per week, for 14 weeks. The 14 weeks of exposure had no adverse effect on the clinical health or growth of rats or mice. Male rats and male mice exposed to 1000 ppm MIBK had a slight but statistically significant increase in liver weight and the liver weight/body weight ratio. Liver weight was also increased slightly in male mice exposed to 250 ppm. No gross or microscopic hepatic lesions related to MIBK exposure were observed. Furthermore, the only microscopic change observed was an increase in the incidence and extent of hyalin droplets within proximal tubular cells of the kidneys of male rats exposed to 250 and 1000 ppm of MIBK. The relevance of the male rat kidney tubular effect to humans is not known. In conclusion, other than the male rat kidney effect, exposure of male and female rats and mice to MIBK at levels up to 1000 ppm for 14 weeks was without significant toxicological effect.

Dimethylethanolamine: acute, 2-week, and 13-week inhalation toxicity studies in rats.

Dimethylethanolamine (DMEA) is a volatile, water-soluble amine that has applications in the chemical and pharmaceutical industries. These studies evaluated the acute and subchronic inhalation toxicity of DMEA. Acute (4-hr) exposures of Wistar rats to DMEA vapor resulted in an LC50 value (95% confidence limits) of 1641 (862-3125) ppm. Clinical signs of nasal and ocular irritation, respiratory distress, and body weight loss were observed in rats exposed to 1668 ppm DMEA and higher. In the 2-week study, F-344 rats exposed to 98, 288, or 586 ppm DMEA for 9 days (6 hr/day) during an 11-day period also exhibited signs of respiratory and ocular irritation (except the 98 ppm group). All animals of the 586 ppm group and 4 of 15 male rats of the 288 ppm group died. Body weight values for the 288 ppm group were reduced to about 75% of preexposure values, while the 98 ppm group gained 35% less weight than controls. Statistically significant differences in clinical pathology parameters (288 ppm group) and in organ weight values (288 and 98 ppm groups) probably resulted from the decreased food consumption and not from specific target organ toxicity. In the groups evaluated histologically (the 98 and 288 ppm groups) the eye and nasal mucosa were the primary target organs. In the 13-week subchronic study, F-344 rats were exposed to 0, 8, 24, or 76 ppm DMEA for 6 hr/day, 5 days/week for 13 weeks. The principal exposure-related changes were transient corneal opacity in the 24 and 76 ppm groups; decreased body weight gain for the 76 ppm group; and histopathologic lesions of the respiratory and olfactory epithelium of the anterior nasal cavity of the 76 ppm group and of the eye of several 76 ppm group females. Rats maintained for a 5-week recovery period only exhibited histological lesions of the nasal tissue, with the lesions being decreased in incidence and severity. DMEA acts primarily as an ocular and upper respiratory tract irritant and toxicant at vapor concentrations of 76 ppm, while 24 ppm or less produced no biologically significant toxicity in rats. Thus, 24 ppm was considered to be the no-observable-effect level.

Respiratory tract changes in guinea pigs, rats, and mice following a single six-hour exposure to methyl isocyanate vapor.

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. The majority of deaths occurred during postexposure days 1 through 3. 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. Body weight losses were common in all species following MIC exposures of 2.4 ppm or greater. Microscopic lesions included acute necrosis of the epithelial lining throughout the respiratory tract in animals that died shortly after exposure, coupled with congestion, edema, and inflammation. A microscopic lesion that appeared unique to guinea pigs was bronchiolitis obliterans (where the products of necrosis and inflammation completely closed the bronchioles). Additional microscopic lesions observed in some animals that died or were sacrificed at the end of the study (postexposure day 14) consisted of squamous metaplasia of respiratory epithelium in the nasal cavity, which extended into the larynx, trachea, and in some cases, the bronchi.(ABSTRACT TRUNCATED AT 250 WORDS)

Methyl isocyanate eight-day vapor inhalation study with Fischer 344 rats.

Groups of ten male and ten female Fischer 344 rats were exposed by inhalation 3.1, 0.6, 0.15, or 0.0 (control) ppm of methyl isocyanate (MIC) vapor 6 hr per day for 8 days (two 4-day sessions separated by a 2-day rest). Evaluation of toxic effects included body weight, food consumption, organ weights, and selected hematologic, ophthalmic, neurologic, gross anatomic, and histologic examinations. There were no deaths during the study. Rats of the 3.1 ppm exposure group had decreased body weights, food consumption, and blood oxygen saturation (males only). An increase in hemoglobin concentration (males only) and in lung weights (absolute and as a percentage of body weight) were also observed in the 3.1 ppm rats. Ophthalmic or neurofunctional behavior evaluations were negative for all MIC exposure groups. Only 3.1 ppm of MIC vapor resulted in lesions in the respiratory tract, 0.6 or 0.15 ppm did not. The types of lesions observed were inflammation and squamous metaplasia in the nasal cavity, trachea, and bronchi; inflammation of the bronchioles and alveoli; and submucosal fibroplasia of the bronchioles. No significant lesions were observed in tissues other than those of the respiratory tract in all MIC exposure groups. The results of this study indicate the current 0.02 ppm threshold limit value for MIC is not too high regarding toxicity.

Eighty-five day postexposure follow-up study in Fischer 344 rats after repeated exposures to methyl isocyanate vapor.

The objectives of this study were to describe the microscopic lesions in the respiratory tract of Fischer 344 rats as a result of 4- or 8-days exposure (6 hr/day) of 3 ppm MIC and to characterize the postexposure development of these lesions up to day 85. All rats survived the exposure regimen, although significant decreases in body weight and encrustation of the eyes, nose, or mouth were observed. During the first 15 days of postexposure, the rats were hypoactive and had increased respiratory rates. Male mortality was as high as 63%; only 5% of the MIC-exposed females died. The cause of death was interpreted to be respiratory compromise complicated by anorexia and probably dehydration as well. During the next 28 postexposure days, 48% of the male survivors died, while only 3% of the female survivors died. Throughout the 85-day postexposure period, body weight gains in the MIC-treated groups were consistently below control values. Inflammatory and squamous metaplastic lesions of the respiratory tract, observed the day following completion of either the 4- or 8-day exposure regimen, decreased in both frequency and/or severity in survivors of the 85-day postexposure period, indicating recovery from the cytotoxic and irritating effects of MIC vapor. The squamous metaplastic epithelium was replaced by regenerative epithelium beginning in the deeper portion of the respiratory tract.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological effects of short-term, high-concentration exposure to methyl isocyanate. I. Study objectives and inhalation exposure design.

Early reports from India indicated that humans were dying within minutes to a few hours from exposure to methyl isocyanate (MIC). Attempts to explain the cause(s) of these rapid mortalities is where Union Carbide Corporation concentrated its post-Bhopal toxicologic investigations. The MIC studies involving rats and guinea pigs focused primarily on the consequences of acute pulmonary damage. All MIC inhalation exposures were acute, of short duration (mainly 15 min), and high in concentration (ranging from 25-3506 ppm). MIC vapors were statically generated in a double chamber exposure design. Precautionary measures taken during exposures are discussed. Guinea pigs were more susceptible than rats to MIC exposure-related early mortality. A greater than one order of magnitude difference was observed between an MIC concentration that caused no early mortality in rats (3506 ppm) and an MIC concentration that caused partial (6%) early mortality in guinea pigs (225 ppm) for exposures of 10 to 15 min duration. For both species, the most noteworthy clinical signs during exposure were lacrimation, blepharospasm, and mouth breathing. Fifteen minute LC50 tests with 14-day postexposure follow-up were conducted, and the LC50 (95% confidence limit) values were 171 (114-256) ppm for rats and 112 (61-204) ppm for guinea pigs. Target exposure concentrations for the toxicologic investigations of MIC-induced early mortality were established. A short summary of pertinent results of Union Carbide Corporation's post-Bhopal toxicologic investigations is presented.

Biological effects of short-term, high-concentration exposure to methyl isocyanate. II. Blood chemistry and hematologic evaluations.

Human, rat, and guinea pig packed erythrocytes exposed to 100, 500, or 1000 ppm of methyl isocyanate (MIC) vapor in vitro showed a concentration-related inhibition of cholinesterase (ChE) activity. Rat and guinea pig packed erythrocytes showed an almost complete inhibition of ChE activity at 2000 ppm. In vitro exposures of human and guinea pig blood to 1000 or 2000 ppm of MIC vapor resulted in qualitative alterations in the electrophoretic mobility of hemoglobin (Hb) as measured by citrated agar electrophoresis. In rats and guinea pigs, neither IV injection of liquid MIC nor in vivo exposure to 1000 ppm of MIC by inhalation resulted in any inhibition of erythrocyte ChE activity or alteration in Hb electrophoretic mobility. As a result of these observations, it was concluded that neither ChE inhibition nor structural alteration of Hb were major contributing factors to death resulting from MIC exposure. Rats and guinea pigs receiving IV injections of liquid MIC showed an increase in creatine kinase (CK) levels. This increase could not be attributed to a specific isoenzyme of CK by ion exchange chromatography. Rats exposed to 100, 600, or 1000 ppm of MIC and guinea pigs exposed to 25, 125, or 225 ppm of MIC and bled immediately following a 15-min exposure or at 1, 2, 4, or 16 hr postexposure had the following alterations in blood parameters: an increase in CK, increases in hemoglobin concentration and hematocrit, reticulocytosis (rats only), neutrophilia, a decrease in blood pH and PO2, and an increase in blood PCO2.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological effects of short-term, high-concentration exposure to methyl isocyanate. III. Influence on gas exchange in the guinea pig lung.

The influence of methyl isocyanate (MIC) inhalation on the gas exchange function of the lungs in guinea pigs was studied by measuring arterial blood gases, pH, and tracheal pressure during constant-volume, artificial ventilation with air or 100% O2 at 40 and 120 min after exposure. A 15 min exposure to MIC at concentrations of 240 to 628 ppm caused a marked reduction in PaO2 and pHa and an elevated tracheal pressure during artificial ventilation. The low PaO2 was only slightly elevated when the animals were ventilated with 100% O2. Although the dry-wet lung weight ratio was reduced at the highest exposure concentration, the effect was not severe and no significant increase in lung water was found at the lower concentrations. MIC inhalation caused severe pulmonary blood shunting and ventilation/perfusion imbalance. This, in turn, led to hypoxemia, metabolic acidosis, and tissue hypoxia, which could produce death. The pulmonary gas exchange deficit likely resulted from bronchial and bronchiolar obstruction caused by sloughed epithelium and other debris from intra- and extrapulmonary airways.

Biological effects of short-term, high-concentration exposure to methyl isocyanate. V. Morphologic evaluation of rat and guinea pig lungs.

The morphologic changes induced in the lungs of rats and guinea pigs exposed to high concentrations of MIC vapor (100, 600, and 1000 ppm in the rat and 25, 125, 225, and 675 ppm in the guinea pig) for a short time (15 min) in a static exposure chamber were evaluated at varying postexposure periods (0, 1, 2, 4, and 16 hr). The 675 ppm-exposed guinea pigs were evaluated only immediately following removal from the chamber. Attention was primarily focused on the intrapulmonary conducting airways and the parenchyma (gas exchange region) of the lungs. The severity of morphologic changes observed by light microscopy was directly correlated with exposure concentration and time postexposure in both species. Specifically, degenerative changes were observed in the bronchial, bronchiolar, and alveolar epithelium in both species. Quantitative differences were observed; 100 ppm of MIC in the rat resulted in much less damage than did 125 ppm of MIC in the guinea pig. Morphologic evidence of sloughing of large sheets of conducting airway epithelium with fibrin buildup and increased mucus production resulted in plugging of major airways and atelectasis. These observations support the hypothesis that tissue hypoxia was a major contributing factor resulting in death.

Reproduction and fertility evaluations in CD rats following nitrobenzene inhalation.

A two-generation reproduction study was performed by exposure of Sprague-Dawley CD rats to concentrations of 40, 10, 1, or 0 (control) ppm of nitrobenzene (NB) vapor. No NB-related effects on reproduction were observed at 10 or 1 ppm. At 40 ppm, a decrease in the fertility index of the F0 and F1 generations occurred, which was associated with alterations in the male reproductive organs. Specifically, weights of testes and epididymides were reduced and seminiferous tubule atrophy, spermatocyte degeneration, and the presence of giant syncytial spermatocytes were observed. The only significant finding in the litters derived from rats exposed to 40 ppm was an approximate 12% decrease in the mean body weight of F1 pups on Postnatal Day 21. Survival indices were unaltered. To examine the reversibility of this selective effect on male gonads, the F1 males from the 40-ppm group were allowed a 9-week nonexposure period and mated to naive females. An almost fivefold increase in the fertility index was observed, indicating at least partial functional reversibility upon removal from NB exposure. Also, the numbers of giant syncytial spermatocytes and degenerated spermatocytes were greatly reduced. The results of this study support the selection of 10 ppm of NB as the no-observable-effect level for reproduction and fertility effects in rats.

Methyl isocyanate subchronic vapor inhalation studies with Fischer 344 rats.

Groups of Fischer 344 rats were exposed to 3.1, 0.6, 0.15, or 0.0 (control) ppm of methyl isocyanate (MIC) vapor 6 hr per day for two 4-day sessions separated by a 2-day rest. There were no deaths during the study. The rats were killed the morning following the last exposure day. The 3.1-ppm-exposed rats had decreased body weight, food consumption, and blood oxygen saturation (males only). Increased hemoglobin concentration (males only) and lung weights were also observed in this group of rats. Multiple histologic lesions, limited to the respiratory tract, were observed in rats of the 3.1-ppm group only. The lesions consisted of necrosis, suppurative inflammation, squamous metaplasia, and intraluminal and submucosal fibroplasia (bronchi and bronchioles only) which extended from the anterior nasal cavity to the terminal bronchioles. In a second study, rats were exposed to 3.0 ppm MIC, 6 hr per day, for either one or two 4-day sessions and sacrificed on postexposure Days 1, 15, 43, and 85. All rats survived the 4- or 8-day exposure regimen, although significant decreases in body weight and encrustation of the eyes, nose, or mouth area were observed. During the first 15 days postexposure, male mortality was 63%; only 6% of the MIC-exposed females died. The cause of death was interpreted to be a combination of pulmonary vascular and inflammatory changes coupled with anorexia. For survivors, recovery from the necrotizing and irritating effects of MIC vapor was observed. Squamous metaplasia of respiratory epithelium, observed in rats sacrificed at the end of the exposure period, was replaced by tall pseudostratified columnar (regenerative) epithelium beginning in the bronchi and bronchioles as well as the distal trachea. Collagen maturation and condensation of the intraluminal and submucosal fibroplasia occurred during the postexposure period. The results of these investigations support the current threshold limit value for MIC of 0.02 ppm.

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.

Acute inhalation studies with methyl isocyanate vapor. II. Respiratory tract changes in guinea pigs, rats, and mice.

Hartley guinea pigs, Fischer-344 rats, and B6C3F1 mice of both sexes were exposed to varying concentrations of methyl isocyanate (MIC) vapor with the highest concentration being 20.4 ppm for rats and mice and 10.5 ppm for guinea pigs. A control group for each species was exposed to air only. All animals were exposed for a duration of 6 hr, and survivors were sacrificed 14 days following exposure. The respiratory tract was removed and examined microscopically from all animals. Guinea pigs were more sensitive to the MIC vapor than were rats which were in turn more sensitive than mice. Gross lesions encountered in many of the animals that died consisted of nasal discharge, often blood tinged, and discoloration of the lungs. Microscopic lesions included acute necrosis of epithelial lining throughout the respiratory tract in animals that died shortly after exposure coupled with congestion, edema, and inflammation. A microscopic lesion which appeared unique to guinea pigs was bronchiolitis obliterans where the necrosis and inflammation had completely closed the bronchioles. Additional microscopic lesions observed in some animals that died or were sacrificed at the end of the study (postexposure Day 14) consisted of squamous metaplasia of respiratory epithelium in the nasal cavity, which extended into the larynx, trachea, and, in some cases, the bronchi. In addition, epithelial regeneration throughout the tract and submucosal fibroplasia in the trachea, bronchi, and bronchioles were observed, the latter lesion being primarily confined to rodents. No animals exposed to 2.4 or 1.0 ppm of MIC vapor died following exposure. There were minimal microscopic lesions at sacrifice in the 2.4 ppm-exposed animals from all three species. Only in guinea pigs were there lesions in the 1.0-ppm group attributed to MIC vapor exposure.