FINAL REPORT STUDY TITLE Acute Inhalation Toxicity (With Histopathology) Study of 1,1,2 Trichloroethane (1, l ,ZTCE) in Rats STUDY DIRECTOR Daniel T. Kirkpatrick, Ph. D., D. A. B. T.

STUDY INITIATED ON December 21,200O STUDY COMPLETED ON October 29,200l PERFORMING LABORATORY WIL Research Laboratories, Inc. 1407 George Road Ashland, OH 44805928 1 LABORATORY STUDY NUMBER WlL 417001 SPONSOR Task Force c/ o Regulatory Sciences International P. O. Box 33 1 Millwood, VA 22314 Page 1 of 311 WILRE~ EARCHLABORATORJES, INC. 1407GEoRGE~ 0m ASHLAND.° H 448059281 (419) 289 8700 ~AX( 419) 289 3650 improving human health and protecting the environment rhrotlglrscimtific research services.@ Task Force Acute Inhalation Toxicity (With Histopathology) Studv of 1 .l .ZTrichloroethane (1 .I .ZTCE) in Rats COMPLIANCE STATEMENT This study, designated WIL 417001, was conducted in compliance with the Environmental Protection Agency Toxic Substances Control Act (EPA TSCA) Good Laboratory Practice Regulations (40 CFR Part 792), the standard operating procedures of WIL Research Laboratories, Inc., and the protocol as approved by the sponsor. A Certificate of Analysis (Appendix A) was supplied by the sponsor; it is unknown whether the characterization analysis was conducted according to Good Laboratory Practices.

Study Director:

2 WIL 417001 Hazardous Air Pollutant Task Force Acute Inhalation Toxicity (With Histopathology) Studv of 1.1.2 Trichloroethane (l. l. ZTCE) in Rats TABLE OF CONTENTS SLllnlllaly Introduction Experimental Design Materials and Methods Observations Statistical Methods Results Conclusion Personnel and Report Submission Quality Assurance Unit Statement References Deviations from the Protocol INDEX OF FIGURES 1: Atmosphere Generation and Exposure System (Low and Mid Concentration Exposure Groups)

2: Atmosphere Generation and Exposure System (High Concentration Exposure Group) &g 6 10 11 13 20 24 25 32 35 36 37 38 l& g 39 40 3 WrL 417001 Hazardous Air Pollutant Task Force 1.

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20. Acute Inhalation Toxicity (With Histopathology) Studv of 1 .1.2 Trichloroethane (1.1 .Z TCE) in Rats INDEX OF TABLES Individual Test Atmosphere Concentrations Summary of Clinical Findings  Total Occurrence/ No. of Animals (Detailed Physical Examinations/ Dispositions)

Summary of Clinical Findings  Total Occurrence/ No. of Animals (Immediately Following Exposure)

Response to Novel Stimulus  Summary of Incidence Body Weights (Grams)  Summary of Means Body Weight Gains (Grams)  Summary of Means Bronchoalveolar Lavage Fluid Values  Summary of Means Gross Necropsy Observations Incidence Summary (Unscheduled Deaths)

Gross Necropsy Observations Incidence Summary (Scheduled Necropsy)

Organ Weights (Grams)  Summary of Means Organ Weights Relative to Final Body Weights (Grams/ 100 Grams)  Summary of Means Histomorphological Diagnosis  Summary Incidence (Unscheduled Deaths)

Histomorphological Diagnosis  Summary Incidence (Scheduled Necropsy)

Individual Clinical Observations (Detailed Physical Examinations/ Dispositions)

Individual Clinical Observations (Immediately Following Exposure)

Individual Response to Novel Stimulus Individual Body Weights (Grams)

Individual Body Weight Gains (Grams)

Individual Bronchoalveolar Lavage Fluid Values Individual Gross and Microscopic Description of Organs (Unscheduled Deaths) Pa. g; e 42 45 52 56 60 64 68 70 71 72 76 80 88 101 110 116 120 136 152 160 4 WIL 417001 Hazardous Air Pollutant Task Force 21.

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G. Acute Inhalation Toxicity (With Histopathology) Study of 1 , 1 .2 Trichloroethane ( 1,1,2 TCE) in Rats JNDEX OF TABLES tndividual Gross and Microscopic Description of Organs (Scheduled Necropsy)

Individual Organ Weights and Final Body Weights (Grams)

tndividual Organ Weights Relative to Final Body Weights (Grams/ 100 Grams)

INDEX OF APPENDICES Certificate of AnaIysis (Sponsor Provided Data)

Serological Screening (BioReliance Corporation)

Pretest Clinical Observations Clinical Pathology Methods, Procedures and References Bronchoalveolar Lavage Cytology Report (Gail L. Walter, D. V. M., D. A. C. V. P., D. A. B. T., Clinical Pathology Consultant)

Results of Phagocytosis and Respiratory Burst Assays (Gary R. Burleson, Ph. D., Burleson Research Technologies)

Study Protocol 166 220 22s ii2422 236 238 242 245 247 264 292 5 WL 417001 Hazardous Air Pollutant Task Force Acute Inhalation Toxicity (With Histopathology) Study of 1 ,1,2 Trichloroethane (1,1,2 TCE) in Rats SUMMARY The toxicological response, including exposure level response relationships, of the test article in rats was evaluated following a single, four hour exposure. The test article was administered to four groups of 10 male and 10 female rats as a vapor via whole body exposure at target concentrations of 0, 50, 200 and 1500 ppm. Five animals/ sex/ group were assigned for anatomical pathology evaluation (gross and microscopic examinations) and five animals/ sex/ group were assigned for evaluation of bronchoalveolar lavage fluid (BALF). For the female BALF evaluation, the target concentration for the high exposure level was decreased to 1000 ppm based on the death of three females following exposure targeted at 1500 ppm. Following exposure, the animals were maintained for approximately 24 hours prior to euthanasia. Body weights were determined and observations for clinical signs of toxicity were conducted at the appropriate intervals for all animals.

Three females in the 1527 ppm group and three females in the 840 ppm group were found dead or euthanized in extremis as a result of test article exposure.

Animals in the high concentration groups (1527, 1474 and 840 ppm) showed no reaction to novel stimulus (a sudden noise) at the approximate midpoint of exposure. All other test article groups reacted similarly to the control group and were considered normal. Immediately following exposure, all animals in the high concentration groups (1527 ppm and 1474 or 840 ppm) appeared to be sleeping, were lethargic, ataxic, had decreased respiration rates and/ or clear discharge of the eyes. These clinical effects observed for high concentration animals during or immediately following exposure were primarily associated with anesthetic or narcotic effects of the test article, a chlorinated, short chain hydrocarbon. On the day following exposure, ataxia, decreased respiration rate, lethargy, hypothermia, reddish brown urine and/ or clear discharge of the eye( s) were observed in the high concentration groups. There were no other toxicologically significant clinical observations immediately following or on the day following exposure.

Test article related body weight effects were also limited to the high concentration groups. Decreased mean body weight gains were observed in the 1527 ppm group males and females and in the 1474 ppm group males and 840 ppm group females on the day following exposure (day 1) when compared to the control group. As a result, mean body weights were decreased in the 1527 ppm group males and females and 1474 ppm group males on day 1. There were no other remarkable changes in body weight data.

6 WIL 417001 Hazardous Air Pollutant Task Force Mean total protein was increased in the bronchoalveolar lavage fluid of the 1474 ppm group males and 840 ppm group females. No microscopic findings in the lung correlated with this increase in lavage fluid total protein. Additionally, no alterations in BALF total or differential cell counts attributed to test article exposure were observed at any concentration. The functional capacity of alveolar macrophages isolated from bronchoalveolar lavage fluid was also evaluated. Phagocytic activity was reduced in all male 1,1,2 TCE exposure groups with the greatest effect in the mid concentration group. For females, phagocytic activity appeared to be decreased only in the mid concentration group. It is noteworthy that there was no exposure concentration dependence for the apparent effects on phagocytic activity in males. In addition, historical control data, which would provide normal or typical values for phagocytic activity of alveolar macrophages from Fischer 344 rats (determined using the same assay method), is not available. The toxicological significance of the changes observed in this study is uncertain based on the lack of exposure concentration dependence and the unavailability of historical control data. There were no test article related effects on respiratory burst activity at any concentration.

For the female rats from the 1527 ppm group that died prior to the scheduled necropsy, the only internal macroscopic finding at necropsy was a dark red area in the glandular portion of the stomach of one animal. At the scheduled necropsy, dark red area( s) on the liver were observed for two males and a pale liver was observed for one female in the 1527 ppm group, Dark red areas in the glandular and/ or non glandular portions of the stomach were observed for one male in the 1527 ppm group and dark red contents in the cecum, ileum, jejunum and/ or stomach were observed for four males and one female in the 1527 ppm group. Dark red contents in the urinary bladder were also observed for one male in the 1527 ppm group. There were no other internal macroscopic findings at the scheduled necropsy. Mean relative (to final body weight) kidney weight in the 1527 ppm group males was significantly (~~ 0.01) higher than the control group. The increase correlated with the microscopic findings noted for the kidney. There were no other test article related effects on organ weights.

Test article related microscopic changes were seen in the liver, nasal cavity, larynx, kidney and stomach. In the liver, centrilobular hepatocellular necrosis (moderate or severe) was observed in all animals in the 1527 ppm group and in 4/ 5 females in the 18 1 ppm group. Chlorinated, short chain hydrocarbons, including 1 ,I ,2 trichloroethane, have been widely reported to produce cell death in the centrilobular areas of the liver. The localization of liver injury appears to be related to the distribution of microsomal metabolizing enzymes. A relatively high concentration of microsomal metabolizing enzymes is also present in the rodent nasal olfactory epithelium. In the current study, minimal to severe necrosis was observed in multiple nasal sections of all 181 and 1527 ppm group males and females with severity tending to increase with exposure level and proximity to the nares. At 58 ppm, all females and 2/ 5 males had minimal to mild 7 w& 417001 Hazardous Air Pollutant Task Force olfactory epithelial necrosis in at least one nasal section. The nasal olfactory epithelium of the rat is extremely sensitive to agents that may be activated by cytochrome P 450 metabolizing enzymes. It has been reported that the fraction of the nasal airways covered by olfactory epithelium is approximately 50% in the F344 rat compared to 3% in the human6. In addition, based on information cited by Haschek and Witschi, the relative surface area of the nasal airways (normalized to body weight) is approximately 16 times greater in the rat compared to the human'. As a result, the rat may be considered one of the most sensitive animal models for this type of airway epithelial lesion. The finding of minimal or mild olfactory epithelial cell necrosis for animals from the 58 ppm group is clearly test article related, but it is considered unlikely to represent a toxicologically adverse response. It is likely that such minor effects involving a small fraction of the olfactory epithelium would completely resolve following a single exposure.

Single cell necrosis of the transitional epithelium was seen in the turbinates and lateral wall of nasal sections II and, variably, IlI from all males and females in the 1527 ppm group. Acute inflammation of the larynx was observed in all females and 3/ 5 males in the 1527 ppm group. Four of five males in the 58 ppm group and 2/ 5 males in the 181 ppm group also had minimal laryngeal inflammation. It was unclear why the highest incidence of this change was observed in the lowest concentration group. The ventral pouch of the larynx was the most commonly affected area. The transitional epithelium of the larynx in the rat has also been found to be extremely sensitive to injury by a broad range of chemical agents6. The finding of minimal laryngeal inflammation with no concentration dependence suggests that this finding has little or no toxicological significance.

A small number of renal cortical tubules in the 1527 ppm group contained a brightly eosinophilic to basophilic coarsely granular material. The location of this luminal debris was nonspecific and the composition and significance of this change was unclear. In the glandular stomach, ulceration and/ or erosion was observed at the 1527 ppm exposure level. This change was not seen in any other groups and was the most likely source of the dark red areas and contents seen in the stomach at necropsy. This change may be secondary to stress and not a direct effect of 1,1,2 TCE exposure. No other test article related microscopic changes were observed.

Based on the microscopic findings (olfactory epithelial necrosis and acute laryngeal inflammation) and effects on phagocytic activity of alveolar macrophages from bronchoalveolar lavage fluid, the no observed effect level (NOEL) for males and females was less than 58 ppm when exposed to 1,1,2 trichloroethane as a single, four hour vapor exposure in albino rats. However, the findings noted for the 58 ppm group animals were of questionable toxicological significance, in particular in relation to human risk. There was no exposure concentration dependence for the apparent decrease in alveolar macrophage phagocytic activity. The respiratory tract tissues affected in this group are known to be extremely sensitive to chemical injury in the rat. Since olfactory epithelium 8 w& 417001 Hazardous Air Pollutant Task Force covers approximately 50% of the rat's nasal airways, minimal to mild lesions are considered unlikely to have an adverse effect on function. Laryngeal inflammation of minimal severity is also unlikely to be toxicologically important. Such minor effects in tissues that are extremely sensitive in the rat compared to other species may be misleading in estimating human risk.

9 wIIZ417001 Hazardous Air Pollutant Task Force Acute Inhalation Toxicity (With Histopathology) Studv of 1.1.2 Trichloroethane (1,1.2 TCE) in Rats INTRODUCTION The objective of this study was to characterize the toxicological response, including exposure level response relationships, of the test article in rats following a single, four hour, whole body vapor exposure. Anatomic pathology and effects on body weight, clinical condition and mortality were evaluated. In addition, to characterize potential adverse effects on the respiratory tract, histopathological examination of fixed tissues and evaluation of cellular and biochemical parameters in bronchoalveolar lavage fluid (BALI!) were performed. Histopathological examination was also performed for the liver, kidneys, adrenals and stomach. The inhalation route is a potential route for human exposure to the test article.

The protocol was designed and the study was conducted in compliance with the following guideline: U. S. Environmental Protection Agency Toxic Substances Control Act (EPA TSCA) guideline 799.9135: Acute Inhalation Toxicity with Histopathology.

lO wIL 417001 Hazardous Air Pollutant Task Force Acute Inhalation Toxicity (With Histopathology) Studv of 1 .1,2 Trichloroethane (1 ,1,2 TCE) in Rats EXPERIMENTAL DESIGN The study consisted of four groups, each consisting of 10 male and 10 female Fischer 344 albino rats. Each group was exposed to the test article for a four hour period. For the mid level exposure on February 25, 2001, the exposure period was extended 54 minutes to compensate for a syringe pump failure that resulted in a period of low atmosphere concentration. Five animals/ sex/ group were assigned for anatomical pathology evaluation (gross and microscopic examinations) and five animals/ sex/ group were assigned for evaluation of bronchoalveolar lavage fluid (BALF). Following exposure, the animals were maintained for approximately 24 hours prior to euthanasia. Body weights were determined and observations for clinical signs of toxicity were conducted at the appropriate intervals for all animals.

The male and female animals for anatomical pathology assessment were exposed on February 25, 2001. The animals for BALF evaluation were exposed on February 26,200l (males) and February 27,200l (females). The target concentration for the high exposure level for the female BALF evaluation was lowered from 1500, ppm to 1000 ppm based on the death of three of five female rats exposed to 1527 ppm on February 25,200l. Data for the anatomical pathology assessment phase were collected in computer protocol WIL 417001 and data for the BALF evaluation phase were collected in computer protocol WL417001A. The dose group designations for the BALI? animals are presented as "male/ female" ppm on the report tables.

The following table summarizes the experimental design:

ll WlL 417001 Hazardous Air Pollutant Task Force Group Number of Target 0 Animals Concentration (pnmj Date of Exuosure W& 417001 0 58 181 1527 5M, 5F 0 February 25,200 1 5M, 5F 50 February 25,200l 5M, 5F 200 February 25,200l 5M, 5F 1500 February 25,200l WL417001A 0 60 205 1474 5M 0 February 26,200l 5M 50 February 26,200 1 5M 200 February 26,200l 5M 1500 February 26,200 1 0 5F 0 February 27,200l 45 5F 50 February 27,200l 177 5F 200 February 27,200l 840 5F 1000* February 27,200l M = Male, F = Female * = The target concentration was lowered due to the death of 3/ 5 females exposed at 1527 ppm on February 25,2001.

Experimental Start Date: February 25,200l Experimental Termination Date: July 12,200l 12 WIL 417001 Hazardous Air Pollutant Task Force MATERIALS AND METHODS Test System:

Rat Strain* *

Justification for Selection:

Source:

Number on Study:

Bodv WeiPht Range:

Acre at Start of Study:

Method of Identification:

Housing:

Quarantine: Fischer 344 This species and strain of animal is recognized to be appropriate for acute inhalation studies.

Charles River Laboratories 401 S. New Hope Road Raleigh, NC 27610 40 males and 40 females. The females were nulliparous and non pregnant.

157 to 197 grams for males and 114 to 137 grams for females at the initiation of exposure Approximately eight weeks old Eat tag Individual suspended wire mesh cages. The animals were maintained by the animal husbandry staff of WIL Research Laboratories, Inc., in accordance with standard operating procedures.

The animals were acclimated to laboratory conditions for a minimum of seven days prior to initiation of exposure.

13 wIL 417001 Hazardous Air Pollutant Task Force Test System (continued):

Disease Screen:

Food and Water:

Environmental Conditions:

Test Article Data:

Identification:

Source: Seven days after receipt, blood was collected from the vena cava of five animals/ sex for serological screening. Serum samples were analyzed for antibodies to the following pathogens: Sendai, RCV/ SDA, Reo, KRV, LCM, H l, PVM, ROPV viruses, M. puhonis and CAR bacillus. The analyses were performed by BioReliance Corporation, Rockville, Maryland (Appendix B). All individual serological assays were negative for all animals evaluated.

PMI Nutrition International, Inc. Certified Rodent LabDiet@ 5002 and reverse osmosis treated municipal water were provided ad libitum. Analysis of feed was performed and provided by the manufacturer. Water was analyzed in accordance with standard operating procedures. Contaminants were not present in feed or water at levels expected to interfere with the objectives of the study. Results of analyses are available upon sponsor request.

Animal room with controlled temperature [70 74° F (21 23"( Z)], humidity (44 50%) and light (12 hours light02 hours dark). Protocol specified ranges for temperature and humidity were 68 76° F (19 25"( Z) and 30 70%, respectively.

1, l ,Ztrichloroethane Dow Chemical 2301 N. Brazosport Blvd. Freeport, TX 7754 1 Date Received: December 29,200O 14 WIL 417001 Hazardous Air Pollutant Task Force Test Article Data (continued):

Lot Number:

Purity:

Stability:

Physical Description:

Storage Conditions: NH06019301 99.56% by weight based on the Certificate of Analysis provided by the supplier (Appendix A). The purity of the test article was also evaluated by the WIL Research Laboratories, Inc. Analytical Chemistry Department and found to be 99% (GLP compliant analysis; data maintained in study records).

Following completion of animal exposures, the stability of the test article was also evaluated by the WIL Research Laboratories, Inc. Analytical Chemistry Department and found to be acceptable (GLP compliant analysis; data maintained in study records).

Clear, colorless liquid Sealed container at ambient temperature Exposure Methods:

The exposures were conducted in four 500 L glass and stainless steel whole body exposure chambers operated under dynamic conditions such that there were at least 12 air changes per hour. One chamber was dedicated to each group. The animals were caged individually, without food and water, during the exposures. Chamber temperature, relative humidity, ventilation rate and negative pressure were monitored continuously and recorded approximately every 30 minutes during exposure. A resistance temperature detector (RTD) and polymeric sensors (Model No. HX94C, Omega Engineering, Inc., Stamford, CT) provided monitoring of temperature and humidity, respectively. Chamber ventilation rate was measured by a sharp edge orifice meter and pressure gauges (Dwyer, Inc., Michigan City, IN) calibrated to convert pressure to airflow rate in liters per minute. Oxygen content of the exposure atmosphere was measured during method development prior to study initiation by a Mine Safety Appliances (Pittsburgh, PA) Remote Sampling System equipped with an oxygen cell. The measured oxygen content was greater than 19%.

15 WrL 417001 Hazardous Air Pollutant Task Force Test Atmosphere Generation Methods:

Vapor atmospheres of the test article were generated using systems similar to those shown in Figures 1 and 2. The test article was delivered at a known rate to the top of a glass vaporization column (Ace Glass, Inc., Vineland, NJ) packed with various sized glass beads. A syringe pump (Harvard Apparatus, Holliston, PA) and appropriately sized glass syringe were used to deliver the test article for the low and mid exposure concentrations. During the mid level exposure on February 25,2001, a syringe pump failure resulted in a period of low atmosphere concentration. To compensate for this excursion, the exposure duration was extended to 4 hours and 54 minutes. A liquid metering pump (Fluid Metering Inc., Oyster Bay, NY) was used for the high exposure concentration. Compressed air at a known flow rate was metered to the bottom of the vaporization column. Vaporization occurred as the test article flowed down through the beads while the compressed air flowed up through the column. Each vaporization column was wrapped with a flexible electric heating tape (Omega Engineering, Inc.) set at approximately 75° C to facilitate vaporization of the test article. The resulting vapor was piped to the exposure chamber inlet where the vapor was diluted to the desired exposure concentration by chamber supply air. The control group received filtered air. Operational conditions for the generation system are provided in the following table:

16 wlL417001 Hazardous Air Pollutant Task Force Group Syringe Size Test Article Compressed Air Chamber or Lab Flow Rate Flow Rate Ventilation liIldh@ i3eaQ Rate WIL 417001 0 58 181 1527 WL417001A Males 0 60 205 1474 WIL 417001A Females 0 45 177 840 NA NA 16 103 20 cc 0.05 17 99 119 50 cc 0.16 0.20 17 119 122 QG20 l/ S" 1.01 14 94 102 NA NA 16 103 20 cc 0.05 17 114 126 50 cc 0.16 17 121 QG20 l/ S" 1.01 14 97 102 NA NA 16 103 20 cc 0.05 17 112 126 50 cc 0.16 0.24 17 115 121 QG20 l/ S" 0.68 1.19 14 96 101 Methods of Characterization of Exnosure Atmospheres:

Nominal Concentration:

Nominal exposure concentrations were calculated as the total weight of test article used during each exposure divided by the total volume of air that passed through the system during the exposure.

Actual Concentrations:

Actual exposure concentrations were measured using gas chromatography. A pump and multi port sampling valve drew samples of the exposure atmospheres to the gas chromatograph (GC). The following table summarizes the GC conditions:

17 WIL 417001 Hazardous Air Pollutant Task Force Instrument: Detector: Column:

Gases Carrier Helium Fuel Hydrogen Air Temperatures (" C) Detector: Column: Injector: Injection Volume Retention Time: Hewlett Packard 5890 Series II Flame ionization DB WAX 0.25 p film thickness130 m x 0.25 mm 80 psig (30 ml/ mm) 16 psig (30 ml/ mm) 35 psig (300 ml/ min)

250 160 250 0.25 ml Approximately 0.77 minutes The GC was standardized using known concentrations of the test article prepared in Tedlar@ gas bags (SK, Inc., Eighty Four, PA). Each standard was prepared by injecting a known volume of test article into a glass vaporization bulb. A continuous flow of air through the bulb carried the vaporized test article to a 40 L bag. A dry test meter (American Meter Co., Horsham, PA) measured the volume of air delivered to the bag. Concentrations of the gas phase standards were calculated as follows:

Concentration (ppm) = VOL x R x T x D x 10m3 x 1 Q 6 LxGMWxP Where: VOL = Volume of the test article, ~1 R= Universal gas constant, 62.36 L mmHg/ mol K T= Nominal laboratory temperature, 294 K D= Density of test article, 1.4350 g/ ml L= Volume of air used to make bag, 30 L GMW= Molecular weight of test article, 133.41 P= Laboratory barometric pressure, 730 mmHg 1( r3 = p. l to ml conversion 106= ppm conversion Standards were prepared that bracketed the target concentration for each exposure. Each standard bag was prepared and analyzed on the GC. An instrument response for each standard was recorded to produce a calibration curve. On the day of exposure, one standard was analyzed to confirm GC calibration. Then for each exposure, the instrument response was converted to concentration from the calibration curve for each sample.

18 WIL 417001 Hazardous Air Pollutant Task Force Temporal Stability and Homogeneity of the Test Atmosphere Homogeneity of the exposure atmospheres was determined during the method development phase of the study. Samples were taken and compared from two remote locations and a reference position. The remote samples were taken from the back upper right and front lower left comers. The reference samples were collected from the approximate center of the chamber. The determinations were performed in triplicate and a mean percent difference from the reference position was calculated for each remote location. Mean concentrations collected at the remote locations were within + 10% of the reference location and considered homogeneous for the purposes of this study. In addition, the stability of the test article concentrations over time was found to be acceptable during method development trial exposures.

Chemical Puritv of the Test Atmosuhere During the 1474 ppm animal exposure, the chemical purity of the test atmosphere was analyzed by the following procedure. A sample of the test atmosphere was analyzed by the gas chromatograph. The GC was optimized and the run length was extended to allow for detection of all test article associated peaks. The high concentration standard (1801 ppm) was then analyzed in the same manner. The percent of total peak area counts represented by the area counts for the atmosphere and standard peaks were calculated and compared. The 1,1,2 TCE peak represented 99.5% and 99.4% of all peaks detected for the test atmosphere and 1801 ppm standard, respectively.

Absence of Aerosol in the Test Atmosnhere During the 1474 ppm animal exposure, the test atmosphere was evaluated for the absence of aerosol using a light scattering aerosol photometer (MIE, Inc. Billerica, MA). The photometer was connected to a strip chart recorder (Omega Engineering, Inc.). The photometer was first standardized with a room air sample. A sample of the test atmosphere was then pulled through the photometer and the output was recorded. No aerosols were detected during this evaluation.

19 WIL 417001 Hazardous Air Pollutant Task Force OBSERVATIONS Mortality:

Each rat was observed for mortality/ moribundity twice daily during the acclimation period, between 3 and 4 p. m. on the day of exposure and before 8 a. m. on the day following exposure.

Detailed Phvsical Examinations:

Detailed physical examinations were performed approximately one week prior to the initiation of exposures, on the day of animals were assigned to groups and on the day following exposure (prior to necropsy). The animals were removed from their home cages and placed in a standard arena for observation. Observations were detailed and carefully noted. Signs noted included, but were not limited to, changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions, and autonomic activity (e. g., lacrimation, piloerection, pupil size, unusual respiratory pattern). Changes in gait, posture and response to handling, as well as the presence of clonic or tonic movements, stereotypes (e. g., excessive grooming, repetitive circling) or bizarre behavior (e. g., self mutilation, walking backwards) were recorded. The absence or presence of findings were recorded for individual animals. If applicable, animals to be eutbanized prior to scheduled necropsy received an additional detailed physical examination prior to euthanasia to assist in determination of the cause of death. Pretest clinical observations are presented in Appendix C.

Clinical Observations:

Each rat was observed immediately following exposure for clinical signs. Observations included, but were not limited to, changes in the skin, fur, eyes and mucous membranes, respiratory, circulatory, autonomic and central nervous system functions, somatomotor activity and behavior patterns. Findings noted at the clinical examination were recorded for individual animals.

At the approximate midpoint of each exposure, visible animals were observed for response to a novel stimulus to determine their state of arousal. The stimulus was produced by allowing a striker (PVC coupling) to contact the glass of the exposure chamber door. The coupling was attached to a length of cotton rope that was held against the glass at approximately 10 cm above the level of the cage rack. The coupling was raised until the rope was approximately perpendicular to the glass and then released. The response of each visible animal was recorded as 1) no reaction, 2) slight reaction, 3) notably increased reaction.

2o WIL 417001 Hazardous Air Pollutant Task Force Body Weights:

Body weights were obtained twice during the acclimation period, prior to exposure and prior to necropsy on the day following exposure.

Necropsy:

Animals were euthanized approximately 24 hours following exposure and underwent a gross necropsy. In addition, complete necropsies were performed on the animals that were found dead. Animals were euthanized by isoflurane anesthesia followed by exsanguinated. At the time of necropsy, the following tissues and organs were collected and preserved in 10% neutral buffered formalin or processed as noted:

Adrenals (2) Aorta Bone with marrow (stemebrae) Brain Forebrain Midbrain Hindbrain Epididymides (2) ' Eyes with optic nerve (2) a Gastrointestinal tract Esophagus Stomach Duodenum Jejunum Ileum Cecum Colon Rectum Heart Kidneys (2) Larynx Liver (sections of two lobes) Lungs (including bronchi, fixed by inflation with fixative at a constant pressure)

a = Placed in Davidson's solution Lymph node Mediastinal Tracheobronchial Nasal Tissues Ovaries with oviducts (2) Pancreas Parathyroidsb Peripheral nerve (sciatic) Pituitary Prostate Salivary glands [mandibular (2)] Seminal vesicles (2) Skeletal muscle (rectus femoris) Skin Spinal cord Cervical Thoracic Lumbar Spleen Testes (2) ' Thymus Thyroids Trachea Urinary bladder Uterus with vagina Gross lesions (when possible)

b = Examined if in the plane of section or when a gross lesion was present c = Placed in Bouin's solution 21 WrL 417001 Hazardous Air Pollutant Task Force Organ Weights The following organs were weighed from the animals at the scheduled necropsy.

Adrenals Brain Kidneys Liver Lungs (prior to inflation with fixative) Ovaries Testes Paired organs were weighed together. Organ to final body weight ratios were calculated.

Microsconic Examination After fixation, protocol specified tissues were trimmed according to the standard operating procedures of WIL Research Laboratories, Inc. Trimmed specimens were processed into paraffin blocks, sectioned at five to eight microns, placed on glass microscope slides and stained with hematoxylin and eosin.

Following collection of the specified tissues, the entire head was removed and decalcified. Six cross sections of the nasal tissues were prepared for microscopic examination in accordance with the method described by Morgan'. This method provides optimal evaluation of the nasopharyngeal meatus and exceeds the requirements of the 799.9135 guideline. The larynx was sectioned to include epithelium covering the base of the epiglottis, the ventral pouch and the medial surfaces of the vocal processes of the arytenoid cartilages.

Microscopic examination of the liver, kidneys, stomach and tissues of the respiratory tract was conducted for all animals in the control and high exposure groups. The respiratory tract tissues examined were the nasal tissues, larynx, trachea, bronchi and lungs, including airways, terminal and respiratory bronchioles, alveolar ducts and sacs and interstitial tissues. Nasal sections II VI, adrenal glands, kidneys, larynx, liver and stomach (non glandular to proximal duodenum) were examined microscopically from all animals in the low and mid exposure groups based on findings in the high exposure groups, on the findings noted for gross lesions and/ or based on organ weights.

Microscopic examination was conducted by Gary Marit, D. V. M., D. A. C. V. P., WIL Research Laboratories, Inc.

22 WJL 417001 Hazardous Air Pollutant Task Force Bronchoalveolar Lavage Evaluation Approximately 24 hours following exposure, surviving animals scheduled for evaluation of bronchoalveolar lavage fluid (BALF) were euthanized by isoflurane and thoroughly exsanguinated. The lungs were examined macroscopically and any lesions were noted. To promote maximum drainage of blood from the lungs, the animals were anesthetized, but not allowed to die prior to initiation of exsanguination. After the diaphragm was punctured to allow the lungs to collapse, the thoracic cavity was carefully opened from the diaphragm to the larynx. A small hole was then cut into the trachea just below the larynx and an appropriately sized ball tipped dosing cannula was tied in place. Each animal was lavaged six times using Hank's Balanced Salt Solution without calcium, magnesium and phenol red. The lavage solution was warmed to 37 + 4° C prior to use. The individual lavage volume was 5 ml for males and 4 ml for females.

Recovered lavage fluids were placed in 15ml (1" lavage) and 50 ml (lavages 2 6) polypropylene centrifuge tubes. The tubes were centrifuged for 10 minutes at 2500 RPM in a refrigerated dentrifuge (2 8° C). Using aseptic met, hods, the supematant from the tubes was decanted into labeled storage tubes. The supematant from the first tube was stored on ice until analyzed for total protein. The cell pellets were resuspended in 4.5 ml of resuspension media (RPMI 1640 with 10% Fetal Calf Serum) using a vortex mixer and then combined. The total volume of the cell suspension was recorded. For total white blood cell count (cells/ ml BALF) and preparation of slides for differential white blood cell count, approximately 0.5 m] of the suspension was transferred to a separate storage tube. Total protein analysis and white blood cell counts were performed by WIL Research Laboratories, Inc. Clinical pathology methods are presented in Appendix D.

Slides for differential count were prepared and shipped to Gail L. Walter, D. V. M., D. A. C. V. P., D. A. B. T, Clinical Pathology Consultant, Kalamazoo, MI, for evaluation (Appendix E). For each differential count, 200 or 201 nucleated cells were counted and identified microscopically as large mononuclear cells, lymphocytes, neutrophils, eosinophils, epithelial cells or mitotic cells.

The remaining cell suspension was used to evaluate the effects of test article exposure on alveolar macrophage function. Phagocytosis and respiratory burst assays were performed using the methods of Burleson' and Rodgers3, respectively. The assay samples were shipped on blue ice via overnight courier to Gary R. Burleson, Burleson Research Technologies, Raleigh, NC, for analysis. The methods and results of the analyses are presented in Appendix F. All Quality Assurance functions for these analyses were the responsibility of Burleson Research Technologies.

23 WIL 417001 Hazardous Air Pollutant Task Force STATISTICAL METHODS All analyses were conducted using two tailed tests for significance levels of 1% and 5% comparing the test article treated groups to the air control group by sex. All means were presented with standard deviations (S. D.) and the numbers of sampling units (N) used to calculate the means. Statistical analyses were not conducted if the number of animals was two or less. All statistical tests were performed using appropriate computing devices or programs. Body weight, body weight change, BALF values and absolute and relative organ weight data were subjected to a one way analysis of variance (ANOVA) followed by Dunnett's test', if the ANOVA revealed statistical significance (~~ 0.05).

24 WIL 417001 Hazardous Air Pollutant Task Force RESULTS Characterization of Exnosure Atmosnheres:

Environmental Conditions:

Mean temperature and relative humidity of the exposure atmosphere are summarized in the following table:

Group !ilzr2@

WIL 417001 0 58 181 1527 Temperature C° Cj Relative Humidity (%) w& Q @ Mean S. D. @

21 0.4 8 47 1.2 8 21 0.5 8 49 2.1 8 24 0.3 9 41 1.6 9 22 0.5 8 46 1.7 8 WIL 4 1700 1 A Males 0 21 0.5 8 43 1.2 8 60 21 0.5 8 44 1.6 8 205 23 0.5 a 40 1.1 8 1474 21 0.0 8 42 1.1 8 WIL 417001A Females 0 21 0.5 8 43 2.3 8 45 21 0.5 8 46 1.8 8 177 23 0.5 8 41 1.8 8 840 21 0.0 9 43 1.9 9 Actual Concentrations (Table 1):

The following table summarizes the actual exposure concentration data for the exposure:

25 WIL 417001 Hazardous Air Pollutant Task Force Group 0 wIL 417001 0 58 181 1527 WL417OOlA Males 0 60 205 1474 WIL 417001A Females 0 45 177 840 Mean Standard 0 0.0 6 58 18.4 8 181 91.4 10 1527 33.0 6 0 0.0 6 60 6.6 6 205 5.3 6 1474 60.1 6 0 0.0 7 45 6.3 7 177 48.1 8 840 185.2 9 Number of SamDIes The calculated tss equilibration time was approximately 18 to 23 minutes for all exposures. To compensate for the equilibration time, the animals were left in the chambers for approximately 20 minutes following shutdown of the generation system at the end of exposure. It should be noted that the mid level exposure on February 25, 2001 (mean concentration of 18 1 ppm) was extended 54 minutes to compensate for a period of low concentrations caused by a syringe pump failure. It can be estimated that the mean exposure concentration was at least 200 ppm (the target exposure level) for approximately a four hour period.

Mortalitv (Tables 2 and 14):

Three females in the `1527 ppm group were found dead on the day following exposure. Three females in the 840 ppm bronchoalveolar lavage (BAL) group died following exposure. For these three females, one was found dead immediately following exposure and one was found dead and one was euthanized in extremis on the day following exposure. Rush (emergency) bronchoalveolar lavage procedures were performed for the two female rats that died or were euthanized on the day following exposure (840 ppm BAL group). There were no other deaths during the study.

26 WIL 417001 Hazardous Air Pollutant Task Force Clinical Observations (Tables 2,3, 14 and 151:

Immediately following exposure, all animals in the 1527 ppm group appeared to be sleeping, had decreased respiration rates and clear discharge of the eyes. Similarly, males in the 1474 ppm BAL group and females in the 840 ppm BAL group appeared to be sleeping, were lethargic and/ or ataxic after exposure. Decreased respiration rates were also observed for one male and four females in these high concentration BAL groups. Clear discharge of the eyes was noted for two males and four females in the 1474 and 840 ppm groups, respectively, immediately following exposure. There were no other toxicologically significant clinical observations immediately following exposure.

On the day following exposure, clinical observations were again limited to animals in the high concentration toxicology/ pathology and BAL groups. Lethargy was observed for 8/ 10 males in the 1527 and 1474 ppm groups combined. Hypothermia (body cool to the touch) was observed for four males in the 1474 ppm BAL group. Decreased respiration rate was noted for three males in the 1527 ppm group and for one male in the 1474 ppm group. In addition, reddish brown urine was noted for 3/ 10 males in the high concentration groups on the day following exposure. Clinical observations in the 1527 ppm group females were clear discharge of the eye( s), decreased respiration rate, hypothermia and lethargy. Ataxia, clear discharge of the eye( s), decreased respiration rate, hypothermia, lethargy and prostration were observed for females in the 840 ppm group. There were no other toxicologically significant clinical observations on the day following exposure.

Response to Novel Stimulus (Tables 4 and 16):

The response to novel stimulus data (from the response induced by a sudden noise) collected at the approximate midpoint of each exposure indicated a definite decrease in the state of arousal in the high concentration groups (1527, 1474 and 840 ppm) when compared to the control group. The responses in the low and mid exposure level groups included a mix of reactions (none, slight and increased) and were considered normal when compared to the control group.

Body Weights (Tables 5,6, 17 and 18):

Mean body weight gains in the 1527 ppm group males and females and in the 1474 ppm group males and 840 ppm group females were significantly (~~ 0.01) lower than the control group on the day following exposure (day 1) when compared to the control group. As a result, mean body weights were significantly (~~ 0.5) lower than the control group in the 1527 ppm group males and females and 1474 ppm group males on day 1. There were no other remarkable changes in body weight data.

27 WIL 41700 1 Hazardous Air Pollutant Task Force Bronchoalveolar Lavage Evaluations (Tables 7 and 19; Appendices E and F):

Mean total protein was significantly (~~ 0.05 or ~~ 0.01) higher than the control group in the bronchoalveolar lavage fluid of the 1474 ppm group males and 840 ppm group females. Relative to the control group means, the mean total protein level was increased 167% and 55% in the high concentration male and female groups, respectively. No alterations in BALF total or differential cell counts attributed to test article exposure were observed.

The test guideline for this acute study requires evaluation of alveolar macrophage function of cells collected by bronchoalveolar lavage. Two assays were used to evaluate macrophage function in this study. Measurement of functional phagocytic activity, by quantifying fluorescence intensity produced by intracellular (i. e., phagocytized) fluorescein labeled Escherichia coli, revealed a decrease in phagocytic activity for male rats exposed to 60, 205 or 1474 ppm 1,1,2 TCE (compared to air exposed controls). However, for female rats, phagocytic activity appeared to be decreased in the mid concentration (177 ppm) exposure group only. It is noteworthy that there was no exposure concentration relationship for the apparent effects on phagocytic activity in males. The magnitude of the effect was similar following exposure to either 60 or 1474 ppm and the most pronounced effect was found in the mid concentration exposure group. In addition, historical control data, which would provide normal or typical values for phagocytic activity of alveolar macrophages from Fischer 344 rats (determined using the same assay method), is not available. The toxicological significance of the changes observed in this study is uncertain based on the lack of exposure concentration dependence and the unavailability of historical control data. The respiratory burst is a metabolic response of phagocytic cells that is associated with microbicidal activity. Exposure to 1,1,2 trichloroethane by inhalation had no effect on respiratory burst activity (neither stimulated nor non stimulated) by alveolar macrophages.

Necropsy (Tables 8,9,20 and 21):

A dark red area in the glandular portion of the stomach was observed for one female in the 1527 ppm group that died early. There were no other internal macroscopic findings for the animals that died prior to the scheduled necropsy.

At the scheduled necropsy, dark red area( s) on the liver were observed for two males and a pale liver was observed for one female in the 1527 ppm group. Dark red areas in the glandular and/ or non glandular portions of the stomach were observed for one male in the 1527 ppm group at the scheduled necropsy. Dark red contents in the cecum, ileum, jejunum and/ or stomach were observed for four males and one female in the 1527 ppm group. Dark red contents in the urinary bladder were also observed for one male in the 28 WIL 417001 Hazardous Air Pollutant Task Force 1527 ppm group. There were no other internal macroscopic findings at the scheduled necropsy.

Organ Weights (Tables 10, 11,22 and 23):

Mean relative (to final body weight) kidney weight in the 1527 ppm group males was significantly (~~ 0.01) higher than the control group. The increase correlated with the microscopic findings noted for the kidney. There were no other test article related effects on organ weights. Mean absolute and relative (to final body weight) adrenal gland weights in the 1527 ppm group males were significantly (~~ 0.01) higher than the control group. However, microscopic examination of the adrenal glands failed to reveal a cause for the increased weights. The change in adrenal gland weight may have been caused by stress. Mean absolute and relative lung weights in the 181 ppm group females were significantly (p& O5 or ~~ 0.01) lower than the control group values. However, since a dose related manner change was not observed, the decreases in lung weights were not considered a result of 1,1,2 TCE exposure. Mean relative brain weight was increased (~~ 0.01) in the 1527 ppm group males. However, the increase was slight and was most likely due to the decrease in mean body weight observed for this group. There were no other remarkable changes in organ weight data.

Microsconic Examination (Tables 12, 13,20 and 21):

Test article related changes were seen in the liver, nasal cavity, larynx, kidney and stomach. In the liver, moderate to severe hepatocellular necrosis was seen in all animals in the 1527 ppm group. Minimal to mild centrilobular necrosis was observed for 4/ 5 females in the 181 ppm group. Centrilobular necrosis was not observed in the 181 ppm group males. The following table summarizes the results for hepatocellular centrilobular necrosis:

In the nasal cavity, test article related changes involved primarily the olfactory epithelium and to a lesser extent, the transitional epithelium. The affected olfactory epithelium was 29 WIL 417001 Hazardous Air Pollutant Task Force generally present in nasal regions III VI. Minimal to severe necrosis was observed in multiple sections of all 181 and 1527 ppm group males and females with severity tending to increase with exposure level and proximity to the nares. At 58 ppm, all females and 2/ S males had minimal to mild olfactory epithelial necrosis in at least one nasal section. Necrosis was generally limited to the dorsal meatus and adjacent septum and ethymoid turbinates. Olfactory epithelial necrosis was not observed in the control group. The following table summarizes the necrosis of olfactory epithelium:

Severity Grade 0 ppm 58 ppm 181 ppm 1527 ppm Necrosis, Olfactory Epitheiium Nasal Level III Males Total n 0 4 5 I Minimal NA NA 1 1 Females , I" z% , L I 1 I Mild NA 1 NA 1 2 1 Moderate NA I I\ rn I 1 I t Severe NA ) NA 1 0 I 2 Total 0 I 1 Minimal NA ; I 1 ; E ; Mild NA n 7 n _.__ 1 Modera! :e NA 0 6 2 , PP., P,, U~" c. IG NA 0 0 3 I I rr+ nl n 7 c z . . I Level IV h, rmlnr 1 Nasal 1 "Ia*

Minimal iA ; ;, J 1 Mild NA 0 5 3 Moderate NA 0 0 1 Total 0 5 5 5 Minimal NA 3 n n J , 1 1 1 Mild 1 NA 1 4 4 In the transitional epithelium of the nose, single cell necrosis was seen on the turbinates and lateral wall of nasal sections II and, variably, III from all males and females in the 1527 ppm group. Minimal single cell necrosis was seen in one control group male and one 181 ppm group female.

3o wIL 417001 Hazardous Air Pollutant Task Force In the larynx, acute inflammation (minimal to mild) was observed in all females and 3/ 5 males in the 1527 ppm group. Four of five males in the 58 ppm group and 2/ 5 males in the 181 ppm group also had minimal laryngeal inflammation. It was unclear why the highest incidence of this change was observed in the lowest concentration group. In females, acute inflammation was seen in l/ 5 animals in the control and 181 ppm group, suggesting a no observe effect level (NOEL) of 181 ppm in females. However, there were animal to animal variations in the laryngeal sections and not every desired landmark was well represented. The ventral pouch was the site most commonly affected with inflammation in treated rats.

In the kidney, a small number of cortical tubules contained a brightly eosinophilic to basophilic coarsely granular material. The location of this luminal debris was nonspecific and the composition and significance of this change was unclear. Luminal debris was seen only in rats exposed to concentrations of 1527 ppm (l/ 5 males, 315 females). In the glandular stomach, ulceration and/ or erosion was seen in 3/ 5 males and 2/ 5 females exposed at 1527 ppm. This change was not seen in any other groups and was the most likely source of the dark red areas and contents seen in the stomach at necropsy. This change may be secondary to stress and not a direct effect of l, l, ZTCE exposure. No other test article related microscopic changes were seen. All other observations were considered spontaneous and/ or incidental, and were unrelated to the administration of 1,1,2 TCE.

31 wIL 417001 Hazardous Air Pollutant Task Force CONCLUSION Three females in the 1527 ppm group and three females in the 840 ppm group were found dead or euthanized in extremis as a result of test article exposure. There were no other deaths during the study.

Animals in the high concentration groups (1527, 1474 and 840 ppm) showed no reaction to novel stimulus (a sudden noise) at the approximate midpoint of exposure. All other test article groups reacted similarly to the control group and were considered normal. Immediately following exposure, all animals in the high concentration groups (1527 ppm and 1474 or 840 ppm) appeared to be sleeping, were lethargic, ataxic, had decreased respiration rates and/ or clear discharge of the eyes. These clinical effects observed for high concentration animals during or immediately following exposure were primarily associated with anesthetic or narcotic effects of the test article, a chlorinated, short chain hydrocarbon. On the day following exposure, ataxia, decreased respiration rate, lethargy, hypothermia, reddish brown urine and/ or clear discharge of the eye( s) were observed in the high concentration groups. There were no other toxicologically significant clinical observations immediately following or on the day following exposure.

Test article related body weight effects were also limited to the high concentration groups. Decreased mean body weight gains were observed in the 1527 ppm group males and females and in the 1474 ppm group males and 840 ppm group females on the day following exposure (day 1) when compared to the control group. As a result, mean body weights were decreased in the 1527 ppm group males and females and 1474 ppm group males on day 1. There were no other remarkable changes in body weight data.

Mean total protein was increased in the bronchoalveolar lavage fluid of the 1474 ppm group males and 840 ppm group females. No microscopic findings in the lung correlated with this increase in lavage fluid total protein. Additionally, no alterations in BALF total or differential cell counts attributed to test article exposure were observed at any concentration. `The functional capacity of alveolar macrophages isolated from bronchoalveolar lavage fluid was evaluated. Phagocytic activity was reduced in all male 1,1,2 TCE exposure groups with the greatest effect in the mid concentration group. For females, phagocytic activity appeared to be decreased only in the mid concentration group. It is noteworthy that there was no exposure concentration dependence for the apparent effects on phagocytic activity in males. In addition, historical control data, which would provide normal or typical values for phagocytic activity of alveolar macrophages from Fischer 344 rats (determined using the same assay method), is not available. The toxicological significance of the changes observed in this study is uncertain based on the lack of exposure concentration dependence and the unavailability of historical control data. There were no test article related effects on respiratory burst activity at any concentration.

32 WIL 417001 Hazardous Air Pollutant Task Force For the female rats from the I527 ppm group that died prior to the scheduled necropsy, the only internal macroscopic finding at necropsy was a dark red area in the glandular portion of the stomach of one animal. At the scheduled necropsy, dark red area( s) on the liver were observed for two males and a pale liver was observed for one female in the 1527 ppm group. Dark red areas in the glandular and non glandular portions of the stomach were observed for one male in the 1527 ppm group and dark red contents in the cecum, ileum, jejunum and/ or stomach were observed for four males and one female in the 1527 ppm group. Dark red contents in the urinary bladder were also observed for one male in the 1527 ppm group. There were no other internal macroscopic findings at the scheduled necropsy. Mean relative (to final body weight) kidney weight in the 1527 ppm group males was significantly (p< O. Ol) higher than the control group. The increase correlated with the microscopic findings noted for the kidney. There were no other test article related effects on organ weights.

Test article related microscopic changes were seen in the liver, nasal cavity, larynx, kidney and stomach. In the liver, centrilobular hepatocellular necrosis (moderate or severe) was observed in all animals in the 1527 ppm group and in 4/ 5 females in the 18 1 ppm group. Chlorinated, short chain hydrocarbons, including 1 ,1,2 trichloroethane, have been widely reported to produce cell death in the centrilobular areas of the liver. The localization of liver injury appears to be related to the distribution of microsomal metabolizing enzymes. A relatively high concentration of microsomal metabolizing enzymes is also present in the rodent nasal olfactory epithelium. In the current study, minimal to severe necrosis was observed in multiple nasal sections of all 181 and 1527 ppm group males and females with severity tending to increase with exposure level and proximity to the nares. At 58 ppm, all females and 2/ 5 males had minimal to mild olfactory epithelial necrosis in at least one nasal section. The nasal olfactory epithelium of the rat is extremely sensitive to agents that may be activated by cytochrome P 450 metabolizing enzymes. It has been reported that the fraction of the nasal airways covered by olfactory epithelium is approximately 50% in the F344 rat compared to 3% in the human6. In addition, based on information cited by Haschek and Witschi, the relative surface area of the nasal airways (normalized to body weight) is approximately 16 times greater in the rat compared to the human7. As a result, the rat may be considered one of the most sensitive animal models for this type of airway epithelial lesion. The finding of minimal or mild olfactory epithelial cell necrosis for animals from the 58 ppm group is clearly test article related, but it is considered unlikely to represent a toxicologically adverse response. It is likely that such minor effects involving a small fraction of the olfactory epithelium would completely resolve following a single exposure.

Single cell necrosis of the transitional epithelium was seen in the turbinates and lateral wall of nasal sections II and, variably, III from all males and females in the 1527 ppm group. Acute inflammation of the larynx was observed in all females and 3/ 5 males in the 33 WIL 417001 Hazardous Air Pollutant Task Force 1527 ppm group. Four of five males in the 58 ppm group and 2/ 5 males in the 181 ppm group also had minimal laryngeal inflammation. It was unclear why the highest incidence of this change was observed in the lowest concentration group. The ventral pouch of the larynx was the most commonly affected area. The transitional epithelium of the larynx in the rat has also been found to be extremely sensitive to injury by a broad range of chemical agents6. The finding of minimal laryngeal inflammation with no concentration dependence suggests that this finding has little or no toxicological significance.

A small number of renal cortical tubules in the 1527 ppm group contained a brightly eosinophilic to basophilic coarsely granular material. The location of this luminal debris was nonspecific and the composition and significance of this change was unclear. In the glandular stomach, ulceration and/ or erosion was observed at the 1527 ppm exposure level. This change was not seen in any other groups and was the most likely source of the dark red areas and contents seen in the stomach at necropsy. This change may be secondary to stress and not a direct effect of 1,1,2 TCE exposure. No other test article related microscopic changes were observed.

Based on the microscopic findings (olfactory epithelial necrosis and acute laryngeal inflammation) and effects on phagocytic activity of alveolar macrophages from bronchoalveolar lavage fluid, the no observed effect level (NOEL) for males and females was less than 58 ppm when exposed to l, l, Ztrichloroethane as a single, four hour vapor exposure in albino rats. However, the findings noted for the 58 ppm group animals were of questionable toxicological significance, in particular in relation to human risk. There was no exposure concentration dependence for the apparent decrease in alveolar macrophage phagocytic activity. The respiratory tract tissues affected in this group are known to be extremely sensitive to chemical injury in the rat. Since olfactory epithelium covers approximately 50% of the rat's nasal airways, minimal to mild lesions are considered unlikely to have an adverse effect on function. Laryngeal inflammation of minimal severity is also unlikely to be toxicologically important. Such minor effects in tissues that are extremely sensitive in the rat compared to other species may be misleading in estimating human risk.

34 mom SUBMISSION Kcv. Car01 A Kopp, B. S. Group Supervisor, Gross Pathology and DcvclopmenmlTticoiogy xaboratory Karen S. Rqan, D. V. M.. D& C. V. P., D. A. B. T. Director, Parhology and Veterbny Medicine hhager II, Isolation Toxi~ obgy Gad Mark D. V. M.. D. AC. V. P.

f StafFPathol& ist Gmp Ibbqt% r, Study &. hlysls anti Reports StadyDimor w Dare WIL 417001 Hazardous Air Pollutant Task Force OUALITY ASSURANCE UNIT STATEMENT Date( s) Findings Date( s) Findings Date( s) of Phase Reported to Reported to Insuection( s) ected Studv Director Jlanazemeq 2l27fOl Test Article DispenselExposure 2/ 27/ 01 3/ 30/ 01 10/ 25,26/ 01 Study Records (Ex I) 10/ 29/ 01 10/ 29/ 01 10/ 26/ 01 Study Records (N l) 10126/ 01 10/ 29/ 01 10/ 26/ 01 Study Records (C 12) 10/ 26/ 01 10/ 29/ 01 10/ 26/ 01 Study Records (H l) 10/ 26/ 01 10/ 29/ 01 10/ 26/ 01 Study Records (P l) 10/ 26/ 01 10/ 29/ 01 10/ 29/ 01 Study Records (A 1) 10/ 29/ 01 10/ 29/ 01 10126lOl Draft Report (BALF Cytology Report) 10/ 26/ 01 10/ 29/ O 1 10/ 26,27/ 01 Draft Report (w/ o BALF Cytolow Report) 10/ 29/ 01 10/ 29/ 01 This study was inspected in accordance with the Good Laboratory Practice Regulations, the standard operating procedures of WIL Research Laboratories, Inc., and the sponsor's protocol and protocol amendment( s) with the following exceptions. Data presented in Appendix B were the responsibility of BioReliance Corporation and the data presented in Appendix F were the responsibility of Burleson Research Technologies. Quality Assurance findings, derived from the inspection( s) during the conduct of the study and from the inspections of the raw data and draft report, are documented and have been reported to the study director. A status report is submitted to management monthly.

This report accurately reflects the data generated during the study. The methods and procedures used in the study were those specified in the protocol, its amendments and the standard operating procedures of WIL Research Laboratories, Inc.

The raw data, the retention sample( s), if applicable, and the final report will be stored in the Archives at WIL Research Laboratories, Inc., or another location specified by the sponsor.

Report Audited By:

A Margaret A. Franklin Date Unit Supervisor, Quality Assurance Report Reviewed By:

f j& d 7 q(~ yfpd d A 1. Heather L Skilton, B. S. Group Supervisor, Quality Assurance Date 36 WIL 417001 Hazardous Air Pollutant Task Force REFERENCES 1.

2.

3.

4.

5.

6.

7. Morgan, Kevin, T. (1991) Approaches to the Identification and Recording of Nasal Lesions in Toxicology Studies. Toxicologic Pathology, Vol. 19, No. 4 (Part 1): 337 35 1.

Burleson, G. R., Fuller, L. B., Menache, M. G. and Graham, J. A. (1987) Poly (I): Poly (C) enhanced Alveolar and Peritoneal Macrophage Phagocytosis: Quantification by a New Method Utilizing Fluorescent Beads. Proceedings of the Society of Experimental Biology and Medicine, Vol. 184: 468476.

Rodgers, K., eds. (1995) Measurement of the Respiratory Burst of Leukocytes for Immunotoxicologic Analysis. In: Methods in Immunotoxicology, eds., Vol. 2: 67 77.

Snedecor, G. W. and Co& ran, W. G. (1980) One way classification; analysis of variance. In: Statistical Methods, Seventh Edition. The Iowa State University Press, Ames, IA, pp. 215 237.

Dunnett, C. W. (1964) New tables for multiple comparisons with a control. Biometrics, 20: 482 491.

Dungworth, D. L., Hahn, F. F. and Nikula, K. J. (1995) Noncarcinogenic Responses of the Respiratory Tract to Inhaled Toxicants. In: Concepts in Inhalation Toxicology, 2" d Edition, McClellan, R. O. and Henderson, R. F., eds., Taylor & Francis, Washington, DC., pp. 533 576.

Haschek, W. M. and Witschi, H. R. (1991) Respiratory System. In: Handbook of Toxicologic Pathology, Haschek, W. M. and Rousseaux, C. G., Academic Press, Inc., pp. 761 827.

37 w& 417001 Hazardous Air Pollutant Task Force DEVIATIONS FROM THE PROTOCOL This study was conducted in accordance with the protocol and protocol amendment( s), if applicable, except for the following:

1. ExDerimental Design. For the animal exposure at 181 ppm on February 25,2001, the exposure period was extended 54 minutes to compensate for a syringe pump failure that resulted in a period of low atmosphere concentration.

2. Test Atmosphere Stability and Homoe; eneitv. Spatial homogeneity was performed for Chambers 2 and 4 after animal exposures on March 2 and 5, 2001, instead of during pre study method development. Homogeneity was not conducted on Chamber 3.

3. Chemical Puritv of the Test Atmosphere. Chemical purity of the high concentration atmosphere was conducted during animal exposures on February 26 and 27, 2001, instead of during pre study method development. In addition, chemical purity was not performed in triplicate as required.

4. Absence of Aerosol in the Test Atmosphere. Verification of the absence of aerosol was performed during animal exposure on February 26, 2001, instead of during pre study method development.

5. Bronchoalveolar Lavage Evaluation. Lavages were performed with Hank's Balanced Salt Solution instead of physiological saline.

6. Bronchoalveolar Lavane Evaluation. Individual lavage volumes were 4 5 ml instead of the specified 5 6 ml.

7. Bronchoalveolar Lavage Evaluation. The study protocol stated that information regarding methods, quality assurance and archiving activities and reporting for the subcontracted alveolar macrophage functional assays would be described in a protocol amendment. Instead, this information was included in the report and study records produced by Burleson Research Technologies, Inc.

These deviations did not negatively impact the quality or integrity of the data nor the outcome of the study.

Study Director 38 wIL 417001 Hazardous Air Pollutant Task Force Figure 1: Atmosphere Generation and Exposure System (Low and Mid Concentration Exposure Groups)

Heated ' Vaporization 2 Column c 500 Liter Chamber 39 WlL 417001 Hazardous Air Pollutant Task Force Figure 2: Atmosphere Generation and Exposure System (High Concentration Exposure Group)

500 Liter Chamber Test Article I L T Reservoir 4o WIL 417001 Hazardous Air Pollutant Task Force Acute Inhalation Toxicity (With Histopathology) Study of 1,1,2 Trichloroethane (1~ 1,2 TCE) in Rats Tables l 23 41 PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 1 ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF l, l, Z TCE IN RATS INDIVIDUAL TEST ATMOSPHERE CONCENTRATIONS PAGE 1 ________________________________________

CONCBNTRATIONS FOR STUDY DAY: 0 EXPOSURE DATE: 25 FEB 01 ________________________________________

GROUP: 0 PPM 58 PPM 181 PPM 1527 PPM ________________________________________

TIME INTO CONC . TIME INTO CONC . TIME INTO CONC . TIME INTO CONC . EXPOSURE fHH: MM) (PPM) EXPOSURE (HH: MM) IPPMM) EXPOSURE (HH: MM) fPPM) EXPOSURE (HH: MM) (PPM) ___________________________________ I____ 00: 38 0. 01: oz 0. 01: 47 0. 02: 35 0. 03: 16 0. 03: 48 0.

___________________ MEAN 0. SD 0.0 WI%) 0.00 N 6 00: 29 54. 00: 57 19. 01: 12 56. 01: 42 78. 02: os 74. 02: 31 66. 03: ll 64. 03: 34 49. 00: 34 212. 00: 23 1506. 00: 53 209. 00: 47 1485. 01: 37 0. 01: 32 1506. 01: 50 27. 02: 22 1538. 02: lO 286. 03: Ol 1563. 02: 26 221. 03: 40 1564. 03: 06 208. 03: 44 219. 04: 13 221. 04: 38 206. .

. ___________________________________________________________________________________ 58. 181. 1527. 18.4 91.4 33.0 32.04 50.55 2.16 8 10 6 .

.

_ 43 PROJECT NO.: WIL 417001A SPONSOR: HAP TASK FORCE TABLE 1 ACUTE INH. TOX (WITH HISTOPATHOLGGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL TEST ATMOSPHERE CONCENTRATIONS PAGE 3 _____________"_"__"""""""~""~"""""~~~"~~"~~~~"~~~~~~~""~"~~~~~~"~~"~~~~"~~"~~"~~~~"""~~"~~~~""~~~"""~~""~~~""~~~"~"~

CONCENTRATIONS FOR STUDY DAY: 0 EXPOSURE DATE: 27 FEE 01 ___________________~"~"""""~"""""~~~~~"~~""~~~""~~""""""~~"~~~""~"~"~~~""""~~~"~~~""~~""~~~~"""~~"""~""~"~""~~~~" "____""""_

GROUP: 0 PPM 60/ 45 PPM 205/ 117 PPM 1474/ 840 PPM _____________"______________________ I__""""""""""""""""""""""""""""""" TIME INTO EXPOSURE (HH: MM) _______ 00: 35 01: 19 01: 39 02: Ol 02: 45 03: 14 03: 45 CONC . (PPM)

0. 0. 0. 0. 0. 0. 0. TIME INTO EXPOSURE (HH: MM) _____________

00: 29 01: 15 01: 34 01: sa 02: 42 03: lO 03: 41 (PPM) "   _ "    "

48. 36. 40. 47. 55. 43. 49. TIME INTO CONC . TIME INTO EXPOSURE (HH: MM) (PPM) EXPOSURE (HH: MM)

00: 46 94. 01: lO 124. 01: 30 164. 01: 55 187. 02: lO 228. 02: 37 226. 03: 07 180. 03: 3a 212. 00: 22 995. 01: 05 664. 01: 27 585. 01: 51 613. 02: 05 761. 02: 33 1071. 03: 04 914. 03: 34 965. 04: 16 989.

________________________________________"~~~"~"~~~~"~~~~~"~~"~"~~~~"~~"~~~"~"~~"~""~~~"~~~~~~~~"""~"

MEAN 0. 45. 177. SD 0.0 6.3 48.1 CV(%) 0.00 13.86 27.20 N 7 7 a (PPM)

840. 185.2 22.06 9 PCHCv4.0 06/ 12/ 2001 R: 10/ 29/ 2001 .

.. TABLE 2 (DETAILED PHYSICAL EXAMINATIONS/ DISPOSITIONS) PROJECT NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS PAGE 1 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS ____ MALE  ____________________ TABLE RANGE: 02 25 01 TO 02 26 01 GROUP: 1 2 3 4 ________________________________________"

NORMAL NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION SCHEDULED EUTHANASIA BEHAVIOR/ CNS LETHARGIC CARDIO PULMONARY RESPIRATION RATE DECREASED EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND RIGHT EYE DRIED RED MATERIAL AROUND NOSE EXCRETA URINE APPEARS REDDISH BROWN IN COLOR ORAL/ DENTAL DRIED RED MATERI?& AROUND MOUTH _____________ s______" ______

l 0 PPM 2 58 PPM 3 181 PPM 101 5 5/ 5 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 ,_____ 4 10/ 5 51 5 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 ______________ 1527 PPM 101 5 5/ 5 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 ____________________ 51 5 41 4 1/ 1 21 2 11 1 l/ 1

.

.

.

. PAGE 2 PROJECT NO.: WIL 417001 SPONSOR: HAp TASK FORCE TABLE 2 (DETAILED PHYSICAL BXAMINATI~ NS/ DI. SP~ SITI~ NS) ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS __ F E ,i, A L E _____ __________________________ I_____________ TABLE RANGE: 02 25 01 TO 02 26 01 GROUP: 1 2 3 4 ________________________________________

NORMAL NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION FOUND DEAD SCHEDULED EUTHANASIA BEHAVIOR/ CNS LETHARGIC BODY/ INTEGUMENT BODY COOL TO TOUCH DRIED RED MATERIAL ON DORSAL HEAD DRIED RED MATERIAL ON FORELIMB( S)

CARDIO PULMONARY RESPIRATION RATE DECREASED EYES/ EARS/ NOSE CLEAR DISCHARGE RIGHT EYE CLEAR DISCHARGE LEFT EYE DRIED RED MATERIAL AROUND RIGHT EYE ___________________ l o PPM 2 58 PPM lO/ 5 01 0 5/ 5 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 _____________________ 3 181 PPM 4 10/ 5 01 0 5/ 5 01 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 _________________~ 1527 PPM o/ 0 s/ 5 o/ 0 01 0 o/ 0 o/ 0 Of 0 01 0 o/ 0 1/ 1 _________ 41 4 31 3 2/ 2 1/ 1 1/ 1 I/ 1 1/ 1 2/ 2 2/ 2 21 2 2/ 2 TABLE 2 (DETAILED PHYSICAL EXAMINATIONS/ DISPOSITIONS) PROJECT NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF l, l, Z TCE IN RATS SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS ____ F E M A L E _____ PAGE 3 ____________________~~ TABLE WINGE: 02 25 01 TO 02 26 01 GROUP: 1 2 3 4 ____________________

EYES/ EARS/ NOSE DRIED RED MATERIAL AROUND LEFT EYE o/ 0 o/ 0 o/ 0 11 1 DRIED RED MATERIAL AROUND NOSE o/ 0 o/ 0 01 0 3/ 3 ORAL/ DENTAL DRIED RED MATERIAL AROUND MOUTH 01 0 o/ 0 o/ 0 2/ 2 ____________________" I. 0 PPM 2 5a PFM 3 181 PPM 4 1527 PPM PCSUV4.0 06/ 18/ 2001 ( I ! ! I ! i ! ( ! I ~ I ! ! ! ! ! / ! ! I ! ! ' !

48 / ! I / ! ! i I I / I t I ! / ! ~ ! ! I I ! I 49 I I ! , / 1 , , ! I ! ! / / I I / / ! ~ I I ! II / ~ / I , I I m 51 TABLE 3 (IMMEDIATELY FOLLOWING EXPOSURE) PROJECT NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS PAGE 1 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS __ MALE __ ________________________________________~

TABLE RANGE: GROUP: ________________________________________

NORMAL NO SIGNIFICANT CLINICAL OBSERVATIONS BEHAVIOR/ CNS ANIMAL SLEEPING CARDIO PULMONARY RESPIRATION RATE DECREASED EYES/ EARS/ NOSE CLEAR DISCHARGE RIGHT EYE CLEAR DISCHARGE LEFT EYE _______________

l o PPM 2 58 PPM 3 181 PPM 02 25 01 1 2 3 4 _______________ II__________________

51 5 o/ 0 01 0 o/ 0 o/ 0 _ 4 5/ 5 5/ 5 o/ 0 Of 0 o/ 0 5/ 5 o/ 0 o/ 0 s/ 5 o/ 0 o/ 0 5/ 5 o/ 0 o/ 0 5/ 5 _________~ 1527 PPM I / I ! ! / ! ' / / I ! / /

53 54 55 I II' I I / I ! ! ! ! ! ! ! ! ! !

56 57 i i ! ! III / ! ! ! ! / ! ! ! / ! ! ! !

I ,

58 TABLE 4 (RESPONSE TO NOVEL STIMULUS) PROJECT NO.: WIL 417001A ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS PAGE 2 SPONSOR: HAP TASK FORCE SUMMARY OF CLINICAL FINDINGS: TOTAL OCCURRENCE/ NO. OF ANIMALS ____ F E M A L 8 ____ ________________________________________"

TABLE RANGE: 02 27 01 GROUP: 1 2 3 4 BEHAVIOR/ CNS NO REACTION TO STIMULUS SLIGHT REACTION NOTABLY INCREASED REACTION NOT VISIBLE __________________ l 0 PPM 2 60/ 45 PPM 3 205/ 177 PPM  21 2 4/ 4 4/ 4 5/ 5 I/ 1 I/ 1 l/ 1 o/ 0 2/ 2 01 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 o/ 0 _______________________________________ 4 1474/ 840 PPM MANUALVl . 0 06/ 20/ 2001 R: 06/ 20/ 2001 .

. 60 I I ! I ! I I ! / II : / ! ! ! 1 I , / ! I / / , I I / / ' ! /

61 I , ! I I ! I I ! I / / I ! I ! / ! / / I I ! ! ! ! / ! !

62 / 1 ! / / ! / ! / I / ! / ! 1 1 ! ! I I ! / ! ! ! ! I / I !

63 64 ,

.IDm m . l 4 .mul r N .u) m t 4 . ri .Nul I+ . t 4 65 I I I I I / / ! I I ! I / / / ! ! I ! I ~ ! ! ! ! ! ! ! ! ! 1 ! ! ~ ! / I / ! I / ! ! ~ ! 1 ! ! ! ! ! ! ! ! / ! I ! ,

67 I / I / ! ! ! ! I / I ! ! / / ! i / / / ! I ! ~ I I ! / I ! / ! ! ! ! ! ! ! / / / !

69 I I ! I / ! ! ! ! I / I t I , I I ! I ! ! ! ) / TABLE 9 (SCHEDULED NECROPSY) PROJECT NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS SPONSOR: HAP TASK FORCE GROSS NECROPSY OBSERVATIONS INCIDENCE SUMMARY PAGE 1 SCHEDULED NECROPSY ________________________________________ _____ M A L E   F E M A L E  GROUP: 12 3 4 12 3 4 ____________________ NUMBER OF ANIMALS IN DOSE GROUP NUMBER OF ANIMALS EXAMINED DAY 1 5 5 5 5 5 5 CECUM DARK RED CONTENTS 0 5 5 0 0 0 0 0 0 0 0 0 0 0 5 5 5 0 0 0 0 0 0 0 0 0 0 0 5 1 0 ILEUM DARK RED CONTENTS 0 1 0 JEJUNUM DARK RED CONTENTS 0 3 0 LIVER DARK RED AREA( S) PALE 0 0 2 0 0 0 SKIN RED MATTING YELLOW MATTING 0 0 2 0 0 0 SPLEEN ACCESSORY 0 0 1 STOMACH DARK RED AREA( S) DARK RED CONTENTS 0 0 1 3 0 0 URINARY BLADDER DARK RED CONTENTS 0 NO SIGNIFICANT CHANGES OBSERVED  ALL EXAMINED TISSUES 5 1 1 0 4 5 5 0 0 0 0 0 0 0 0 0 0 0 5 5 5 0 0 0 0 0 1 0 0 0 0 0 4 5 2 0 0 0 0 1 2 1 0 0 1 0 0 ________________________________________

l 0 PPM 2 58 PPM 3 181 PPM 4 1527 PPM PGRSIv4.02 06/ 12/ 2001 I / ! ! I ! ! ! ! ! / I I / I I I / I / ! I I ! / /

73 , , ,

, , I ,

I I 4 I I I 1 I 4 I I 1 1 1 , , I , I I I , I I 75 ! / ! ! ! ! ! I ! ! / ) / ! ! ! / I I 1 1 ! ! / 1 1 1 I 76 ! I ! ! ! ! ! ! ! I ! ! I / I / I I ! / I I ! I ! / / ! / ! ! ! ! ! ! ! ! ! ! / / / ,, I / / ! I i ! ~ / / !

78 79 I / II / 1 / ! I I ! ! ! ! ! I I / / ~ ! ! ! / I / I / /

m u lo Loo mo 00 522 ti2B 00 00 P$

PC2 00 00 BB PP 00 00 BB 852 OO 00 Pb Li2B 00 00 ciE2 Bb 00 80 81 , I , I t ,

82 TABLE 12 (UNSCHEDULED DEATHS) PROJECT NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS PAGE 4 SPONSOR: HAP TASK FORCE HISTOMORPHOLOGICAL DIAGNOSIS  SUMMARY INCIDENCE ____ FE E: __ ________________________________________ 1 2 3 4 . ___________________ GROUP: _______________________________________ NUMBER OF ANIMALS IN DOSE GROUP NUMBER OF ANIMALS EXAMINED 5 0 5 0 5 5 0 3 ADRENAL CORTEX TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE ADRENAL MEDULLA TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE KIDNEYS TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NECROSIS, TUBULAR MINIMAL LUMINAL DEBRIS MINIMAL LIVER TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NECROSIS, HEPATOCELLULAR, CENTRILOBULAR SEVERE 0 NA NA 3 0 NA NA 3 0 NA NA 3 0 NA NA 3 0 NA NA 3 0 NA NA 1 0 NA NA 1 NONE NA NA 1 0 NA NA 2 NONE NA NA 2 0 0 0 NONE NA NA 3 NA NA 0 NA NA 3 NA NA 3 ___  ________________________________________ l 0 PPM 2 5a PPM 3 la1 PPM 4 1527 PPM NA = NOT APPLICABLE 84 I ! I I ! ! I I / I ! ! ! I I ! ! ! ! ! ! ~ /

; , I m 85 TABLE 12 (UNSCHEDULED DEATHS) PROJECT NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLGGY) STUDY OF 1,1,2 TCE IN RATS PAGE 7 SPONSOR: HAP TASK FORCE HISTOMORPHOLOGICAL DIAGNOSIS  SUMMARY INCIDENCE ___ FFJq& LE  ____________________ ____________________________ i_______.. GROUP: 1 2 3 4 ________________________________________  I NUMBER OF ANIMALS IN DOSE GROUP 5 5 5 5 NUMBER OF ANIMALS EXAMINED 0 0 0 3 NASAL LEVEL IV TOTAL NUMBER EXAMINED 0 NA NA 3 EXAMINED, UNREMARKABLE 0 NA NA 0 NECROSIS, OLFACTORY EPITHELIUM 0 NA NA 3 MILD NONE NA NA 1 MODERATE NONE NA NA 1 SEVERE NONE NA NA 1 NASAL LEVEL V TOTAL NUMBER EXAMINED 0 NA NA 3 EXAMINED, UNREMARKABLE 0 NA NA 0 NECROSIS, OLFACTORY EPITHELIUM 0 NA NA 3 MINIMAL NONE NA NA 1 MILD NONE NA NA 2 NASAL LEVEL VI TOTAL NUMBER EXAMINED 0 NA NA 3 EXAMINED, UNREMARKABLE 0 NA NA 0 NECROSIS, OLFACTORY EPITHELIUM 0 NA NA 3 MINIMAL NONE NA NA 1 MILD NONE NA NA 2 ________________________________________________________________________________________~~~~~~________~__________ l 0 PPM 2 58 PPM 3 181 PPM 4 1527 PPM NA = NOT APPLICABLE 87 88 I ,

1 I , , 1 1 1 1 I I / ,

1 I / I , ! I ! ! ! ~ I I I / ! / / I ! ! ! / ! ! / / : :

w 4 g; , , , , I , I ( , ' I PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 13 (SCHEDULED NECROPSY) ACUTE INH. TOK. (WITH HISTOPATHOLOGY) STUDY 0~ 1.1,2 TCE IN RATS PAGE 5 HISTOMORPHOLOGICAL DIAGNOSIS  SUMMARY INCIDENCE  E ___

________________________________________"

GROUP: 1 ,_________________

5 5 2 __________________ NUMBER OF ANIMALS IN DOSE GROUP NUMBER OF ANIMALS EXAMINED DAY 1 URINARY BLADDER 5 5 3 . 5 5 TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NA NA NA NA NA NA NASAL LEVEL I TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE 5 NA NA 5 NA NA NASAL LEVEL II TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NECROSIS, SINGLE CELL MINIMAL MILD INFLAMMATION, SUBACUTE MINIMAL NECROSIS, RESPIRATORY EPITHELIUM MINIMAL METAPLASIA, SQUAMOUS MINIMAL 5 2 0 NONE NONE 2 3 2 3 0 0 NONE NONE 0 1 NONE 1 5 5 0 NONE NONE 0 NONE 0 NONE 0 NONE 4 . 5 5 1 1 5 5 5 0 5 2 3 0 NASAL LEVEL III TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NECROSIS, SINGLE CELL MINIMAL MILD 5 5 5 5 5 5 1 0 0 0 0 3 NONE NONE NONE 1 NONE NONE NONE 2 NONE 1 1 0 NONE ________________________________________ l 0 PPM 2 58 PPM 3 181 PPM 4 1527 PPM NA = NOT APPLICABLE I ! / 1 ! ! ! ! I ! ! ! ! ! / ~ ! ! I ! / I 1 ! ! ! I 1 ! ! I f / 1 / / ! / ! / ! / ! I ! ! ~~ / ! I / / / / / / / ' / 1 / ! PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 13 (SCHEDULED NECROPSY) ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS HISTOMORPHOLCGICAL DIAGNOSIS  SUMMARY INCIDENCE PAGE 8  FEMALE __

__________________ C__________ L_________"~

_________________

NUMBER OF ANIMALS IN DOSE GROUP NUMBER OF ANIMALS EXAMINED DAY ADRENAL CORTEX TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NECROSIS MINIMAL ADRENAL MEDULLA TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE DUODENUM TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE HYPERPLASIA, EPITHELIAL MILD INFLAMMATION, ACUTE MINIMAL KIDNEYS TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NEPHROPATHY MINIMAL INFLAMMATION, CHRONIC MINIMAL GROUP: 1 ______________~

5 5 5 NA NA 2 5 NA NA 1 0 NA NA 1 NONE NA NA 1 5 NA NA 2 5 NA NA 2 NA NA NA 1 NA NA NA 0 NA NA NA 1 NA NA NA 1 NA NA NA 1 NA NA NA 1 2 3 4 .__ L__________ 5 5 5 5 5 2 5 5 2 5 5 1 0 0 0 NONE NONE NONE 0 0 0 NONE NONE NONE ________________________________________ l o PPM 2 58 PPM 3 181 PPM 4 1527 PPM NA = NOT APPLICABLE PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 13 (SCHEDULED NECROPSYI ACUTE INH. TOX. (WITH HISTOPATHOLGGY) STUDY OF 1,1,2 TCE IN RATS HISTOMORPHOLOGICAL DIAGNOSIS  SUMMARY INCIDENCE __ FEMALE      PAGE 9 ____________________~ GROUP: 1 2 3 4 _______________________________________ NUMBER OF ANIMALS IN DOSE GROUP NUMBER OF ANIMALS EXAMINED DAY 1 KIDNEYS  CONTINUED LUMINAL DEBRIS MINIMAL LIVER TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NECROSIS, HEPATOCELLULAR, CENTRILGBULkR MINIMAL MILD MODERATE INFILTRATE, MONONUCLEAR MINIMAL INFLAMMATION, SUBACUTE MINIMAL MILD LARYNX TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE EDEMA MINIMAL MILD INFLAMMATION, ACUTE MINIMAL MILD ___________________~ l 0 PPM 2 58 PPM 3 181 PPM 5 5 5 1 0 NONE NONE NONE 4 4 0 NONE NONE 5 3 1 1 NONE 2 2 NONE 4 1527 PPM 5 5 0 0 NONE NONE il NONE NONE NONE 4 4 0 NONE NONE 5 4 0 NONE NONE 1 I NONE __ c___________ 5 5 5 2 0 0 4 2 3 NONE 1 NONE NONE 2 1 0 1 NONE 3 0 1 NONE 2 NONE 5 2 4 0 0 1 NONE NONE NONE 1 1 2 1 NONE NONE 2 5 2 1 1 I I / ! / ! ! ! I ! ! I ! ! ! ! ! ' / ! ! ! ! ! ! I / / I I I PAGE 11 PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 13 (SCHEDULED NECROPSY) ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY 0~ l, l, z~ C~ IN RATS HISTOMORPHOLOGICIU DIAGNOSIS  SUMMARY INCIDENCE _____ F&,& Z& E ____ ________________________________________

GROUP: 1 2 3 4 ___________________

NUMBER OF ANIMALS IN DOSE GROUP NUMBER OF ANIMALS EXAMINED DAY 1 5 5 5 5 5 5 5 2 TRACHEA TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NASAL LEVEL I TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NASAL LEVEL II TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NECROSIS, SINGLE CELL MINIMAL MILD INFLAMMATION, SUBACUTE MINIMAL METAPLASIA, SQUAMOUS MINIMAL EXUDATE MINIMAL NASAL LEVEL III TOTAL NUMBER EXAMINED EXAMINED, UNREMARKABLE NECROSIS, SINGLE CELL MINIMAL MODERATE ____________________

l 0 PPM 2 58 PPM 3 NA = NOT APPLICABLE 5 NA NA 2 5 NA NA 2 5 NA NA 2 5 NA NA 2 5 5 5 2 3 2 3 0 0 0 1 2 NONE NONE 1 1 NONE NONE NONE 1 2 1 0 0 2 1 NONE NONE 1 2 1 0 1 2 1 NONE 0 0 1 0 NONE NONE 1 NONE 5 5 5 2 4 3 1 0 0 0 0 2 NONE NONE NONE 1 NONE NONE NONE 1 _________________~ 181 PPM 4 1527 PPM / i / / : I I i I ! I I / 1 ! ! I ! ! ! ! ; m TABLE 14 (DETAILED PHYSICAL EXAMINATIONS/ DISPOSITIONS) PROJECT'NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS PAGE 1 SPONSOR: HAP TASK FORCE INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 TO 02 26 01 ______________________________________^_

ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS ________________________________________*

61409 M 0 PPM NORMAL 02 25 01 7: 49 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 24 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61409 M 0 PPM DISPOSITION 02 26 01 13: 05 P SCHEDULED EUTHANASIA 61410 M 0 PPM NORMAL 02 25 01 7: 49 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 24 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61410 M 0 PPM DISPOSITION 02 26 01 13: 05 P SCHEDULED EUTHANASIA 61419 M 0 PPM NORMAL 02 25 01 7: 50 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7~ 24 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61419 M 0 PPM DISPOSITION 02 26 01 13: 06 P SCHEDULED EUTHANASIA 61432 M 0 PPM NORMAL 02 25 01 7: SO P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 25 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61432 M 0 PPM DISPOSITION 02 26 01 13: 06 P SCHEDULED EUTHANASIA 61434 M 0 PPM NORMAL 02 25 01 7: 50 P NO SIGNIFICANT CLINICAL QBSERVATIONS 02 26 01 7: 25 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61434 M 0 PPM DISPOSITION 02 26 01 13: 06 P SCHEDULED EUTHANASIA 61396 M 58 PPM NORMAL 02 25 01 7: 51 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7~ 25 F NO SIGNIFICANT CLINICAL OBSERVATIONS 61396 M 58 PPM DISPOSITION 02 26 01 13: 06 P SCHEDULED EUTHANASIA 61405 M 58 PPM NORMAL 02 25 01 7: 51 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 26 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61405 M 58 PPM DISPOSITION 02 26 01 13: 06 P SCHEDULED EUTHANASIA 61413 M 58 PPM NORMAL 02 25 01 7: 51 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 28 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61413 M 58 PPM DISPOSITION 02 26 01 13: 0 l P SCHEDULED EUTHANASIA 61416 M 58 PPM NORMAL 02 25 01 7~ 51 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7~ 28 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61416 M 58 PPM DISPOSITION 02 26 01 13: 07 P SCHEDULED EUTHANASIA 61431 M 58 PPM NORMAL 02 25 01 7: 52 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 29 P NO SIGNIFICANT CLINICAL OBSERVATIONS __________________ I_______________________________________________.___ GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PROJECT NO.: WIL 417001 SPONSOR: RAP TASK FORCE TABLE 14 (DETAILED PHYSICAL EXAMINATI~ NS/ DISP~ SITIONS) ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 TO 02 26 01 PAGE 2 ______________________________________~ ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS ____________________

61431 M 58 PPM 61399 M 181 PPM 61399 M 61411 M 181 PPM 181 PPM 61411 M 61441 M 181 PPM 181 PPM 61441 M 61442 M 181 PPM 181 PPM 61442 M 61443 M 191 PPM 181 PPM 61443 M 61397 M 61397 M 61397 M 61397 M 61397 M 61406 M 61406 M 61406 M 61408 M 61408 M 61408 M 61408 M 61421 M 191 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM 1527 PPM DISPOSITION 02 26 01 13: 07 P SCHEDULED EUTHANASIA NORMAL 02 25 01 7: 52 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 30 P NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 26 01 13: 07 P SCHEDULED EUTHANASIA NORMAL 02 25 01 7: 52 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 31 P No SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 26 01 13: 07 P SCHEDULED EUTHANASIA NORMAL 02 25 01 7: 53 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 32 P NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 26 01 13: 08 P SCHEDULED EUTHANASIA NORMAL 02 25 01 7: 53 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7~ 32 P NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 26 01 13: Os P SCHEDULED EUTHANASIA NORMAL 02 25 01 7: 53 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 33 P NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 26 01 13: 08 P SCHEDULED EUTHANASIA NORMAL 02 25 01 7: 54 P NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 26 01 13: 08 P SCHEDULED EUTHANASIA BEHAVIOR/ CNS 02 26 01 7: 34 P LETHARGIC CARDIO PULMONARY 02 26 01 7: 35 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 26 01 7: 34 P DRIED RED MATERIAL AROUND NOSE NORMAL 02 25 01 7: 54 P NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 26 01 13: OB P SCHEDULED EUTHANASIA BEHAVIOR/ CNS 02 26 01 7~ 36 P LETHARGIC NORMAL 02 25 01 7: 54 P NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 26 01 13: 08 P SCHEDULED EUTHANASIA BEHAVIOR/ CNS 02 26 01 7: 38 P LETHARGIC CARDIO PULMONARY 02 26 01 7~ 38 P RESPIRATION RATE DECREASED NORMAL 02 25 01 7: 54 P NO SIGNIFICANT CLINICAL OBSERVATIONS ________________________________________ GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT ___________________""""""""

GROUP CATEGORY DATE TIME GRADE OBSERVATIONS .__ ________________________________________"" ANIMAL SEX __

61421 M 61421 M 1527 PPM 1527 PPM 61421 M 1527 PPM 61439 M 1527 PPM 61439 M 1527 PPM 61439 M 1527 PPM 61439 M 1527 PPM 61439 M 1527 PPM 61446 F 0 PPM 61446 F 61452 F 0 PPM 0 PPM 61452 F 61453 F 0 PPM 0 PPM 61453 F 61456 F 0 PPM 0 PPM 61456 F 61491 F 0 PPM d PPM 61491 F 61467 F 0 PPM 58 PPM 61467 F 61470 F 58 PPM 58 PPM DISPOSITION 02 26 03 13: 08 EYES/ BARS/ NOSE 02 26 01 7: 38 02 26 01 7: 39 ORAL/ DENTAL 02 26 01 7: 38 NORMAL 02 25 01 7: 55 DISPOSITION 02 26 01 13: 08 BEHAVIOR/ CNS 02 26 01 7: 40 CARDIO PULMONARY 02 26 01 7: 40 EXCRETA 02 26 01 7: 44 NORMAL 02 25 01 7: 56 02 26 01 7: 44 DISPOSITION 02 26 01 13: 09 NORMAL 02 25 01 7: 57 02 26 01 7: 45 DISPOSITION 02 26 01 13: 09 NORMAL 02 25 01 7: 57 02 26 01 7: 45 DISPOSITION 02 26 01 13: 09 NORMAL 02 25 01 7: 57 02 26 01 7: 45 DISPOSITION 02 26 01 13: 09 NORMAL 02 25 01 7: sa 02 26 61 7: 46 DISPOSITION 02 26 01 13: 09 NORMAL 02 25 01 7: sa 02 26 01 7: 46 DISPOSITION 02 26 01 13: 09 NORMAL 02 25 01 7: 58 02 26 01 7: 46 ___________________ P P P P P P P P P P P P P P P P P P P P P P P P P P P P P _ SCHEDULED EUTHANASIA DRIED RED MATERIAL AROUND NOSE DRIED RED MATERIAL AROUND RIGHT EYE DRIED RED MATERIAL AROUND MOUTH NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED EUTHANASIA LETHARGIC RESPIRATION RATE DECREASED URINE APPEARS REDDISH BROWN IN COLOR NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED EUTHANASIA NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED EUTHANASIA NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED EUTHANASIA NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED EUTHANASIA NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED EUTHANASIA NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED EUTHANASIA NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS ___""__""___""""""" PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 14 (DETAILED PHYSICAL RxAMINATIONS/ DISP~ SITI~ NS) ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1.2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 TO 02 26 01 PAGE 3 GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 14 (DETAILED PHYSICAL ExAMINATION~/ DI~ P~ SITI~ NS) ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 TO 02 26 01 PAGE 4 ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS ____________ __________ __________________________________ I___

61470 F 58 PPM DISPOSITION 02 26 01 13: 09 P SCHEDULED EUTHANASIA 61472 F 58 PPM NORMAL 02 25 01 7: 59 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 61 7: 47 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61472 F 58 PPM DISPOSITION 02 26 01 13: 09 P SCHEDULED EUTHANASIA 61475 F 58 PPM NORMAL 02 25 01 7: 59 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 47 P NO SIGNIFICANT CLIN1CAL OBSERVATIONS 61475 F 58 PPM DISPOSITION 02 26 01 13: lO P SCHEDULED EUTHANASIA 61477 F 50 PPM NORMAL 02 25 01 7: 59 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 48 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61477 F 56 PPM DISPOSITION 02 26 01 13: lO P SCHEDULED EUTHANASIA 61449 F 181 PPM NORMAL 02 25 01 7: 59 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 48 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61449 F 181 PPM DISPOSITION 02 26 01 13: lO P SCHEDULED EUTHANASIA 61474 F 181 PPM NORMAL 02 25 01 8: OO P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 03 7: 49 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61474 F 181 PPM DISPOSITION 02 26 01 13: lO P SCHEDULED EUTHANASIA 61482 F 181 PPM NORMAL 02 25 01 8: 00 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 49 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61482 F 181 PPM DISPOSITION 02 26 01 13: lO P SCHEDULED EUTHANASIA 61486 F I81 PPM NORMAL 02 25 01 8: OO P NO SIGNIFICANT CLINICAL OBSERVATIONS 61486 F 181 PPM DISPOSITION 02 26 01 13: lO P SCHEDULED EUTHANASIA 61486 F 181 PPM EYES/ EARS/ NOSE 02 26 01 7: 49 P DRIED RED MATERIAL AROUND RIGHT EYE 61493 F 181 PPM NORMAL 02 25 01 8: OO P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 7: 50 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61493 F l81 PPM DISPOSITION 02 26 01 13: lO P SCHEDULED EUTHANASIA 61451 F 1527 PPM DISPOSITION 02 26 01 8: 03 P FOUND DEAD 61462 F 1527 PPM NORMAL 02 25 01 8: Ol P NO SIGNIFICANT CLINICAL OBSERVATIONS 61462 F 1527 PPM DISPOSITION 02 26 01 12: 44 P FOUND DEAD 61462 F 1527 PPM BEHAVIOR/ CNS 02 26 01 7: 53 P LETHARGIC _______________________________________"

GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT TABLE 14 (DETAILED PHYSICAL EXAMINATIONS/ DISPOSITIONS) PROJECT NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLGGYI STUDY OF 1,1,2 TCE IN RATS PAGE 5 SPGNSOR: HAP TASK FORCE INDIVIDUAL CLINLCAL OBSERVATIONS TABLE RANGE: 02 25 01 TO 02 26 01 _______________________________________~

ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS ________________________________________

61462 F 1527 PPM BODY/ INTEGUMENT 02 26 01 7: 53 P BODY COOL TO TOUCH 61462 F 1527 PPM CARDIO PULMONARY 02 26 01 7: 53 P RESPIRATION RATE DECREASED 61462 F 1527 PPM EYES/ EARS/ NOSE 02 26 01 7: 52 P DRIED RED MATERIAL AROUND NOSE 02 26 01 7: 52 P CLEAR DISCHARGE RIGHT EYE 02 26 01 7: 52 P CLEAR DISCHARGE LEFT EYE 02 26 01 7: 52 P DRIED RED MATERIAL AROUND RIGHT EYE 61476 F 1527 PPM NORMAL 02 25 01 8: Ol P NO SIGNIFICANT CLINICAL OBSERVATIONS 61476 F 1527 PPM DISPOSITION 02 26 01 13: ll P SCHEDULED EUTHANASIA 61476 F 1527 PPM BODY/ INTEGUMENT 02 26 01 7: 55 P DRIED RED MATERIAL ON DORSAL HEAD 02 26 01 7: 56 P DRIED RED MATERIAL ON FORELIMB( S) 61476 F 1527 PPM CARDIO PULMONARY 02 26 01 7~ 56 P RESPIRATION RATE DECREASED 61476 F 1527 PPM EYES/ EARS/ NOSE 02 26 01 7: 54 P DRIED RED MATERIAL AROUND NOSE 02 26 01 7: 54 P DRIED RED MATERIAL AROUND RIGHT EYE 02 26 01 7: 54 P DRIED RED MATERIAL AROUND LEFT EYE 61476 F 1527 PPM ORAL/ DENTAL 02 26 01 7~ 55 P DRIED RED MATERIAL AROUND MOUTH 61481 F 1527 PPM NORMAL 02 25 01 8: 02 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61481 F 1527 PPM DISPOSITION 02 26 01 7: 16 P FOUND DEAD 61484 F 1527 PPM NORMAL 02 25 01 8: 02 P NO SIGNIFICANT CLINICAL OBSERVATIONS 61484 F 1527 PPM DISPOSITION 02 26 01 13: ll P SCHEDULED EUTHANASIA 61484 F 1527 PPM EYES/ EARS/ NOSE 02 26 01 7: 57 P DRIED RED MATERIAL AROUND NOSE 02 26 01 7~ 57 P CLEAR DISCHARGE LEFT EYE 02 26 01 7: 57 P CLEAR DISCHARGE RIGHT EYE 61484 F 1527 PPM ORAL/ DENTAL 02 26 01 7: 5 l P DRIED RED MATERIAL AROUND MOUTH ______________________________________ GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PCRW4.01 06/ 18/ 2001 PROJECT NO.: WIL 417001A SPONSOR: HAp TASK FORCE TABLE 14 (DETAILED PHYSICAL EXAMINATIONS/ DISPOSITIONS) ACUTE INH. TOX (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 27 01 TO 02 27 01 PAGE 1 ANIMAL SEX GROUP CATEGORY __________________ 61400 M 0 PPM NORMAL 61400 M 0 PPM DISPOSITION 61403 M 0 PPM NORMAL 61403 M 0 PPM DISPOSITION 61404 M 0 PPM NORMAL 61404 M 0 PPM DISPOSITION 61414 M 0 PPM NORMAL 61414 M 0 PPM DISPOSITION 61420 M 0 PPM NORM?& 61420 M 0 PPM DISPOSITION 61395 M 60/ 45 PPM NORMAL 61395 M 60/ 45 PPM DISPOSITION 61398 M 60/ 45 PPM NORMAL 61398 M 60/ 45 PPM DISPOSITION 61425 M 60/ 45 PPM NORMAL 61425 M 60145 PPM DISPOSITION 61430 M 60/ 45 PPM NORMAL 61430 M 60145 PPM DISPOSITION 61435 M 60/ 45 PPM NORMAL 61435 M 60/ 45 PPM DISPOSITION 61407 M 205/ 177 PPM NORMAL 61407 M 205/ 177 PPM DISPOSITION 61415 M 205/ 177 PPM NORMAL 61415 M 205/ 177 PPM DISPOSITION 61422 M 205/ 17? PPM NORMAL 61422 M 205/ 177 PPM DISPOSITION 61424 M 205/ 177 PPM NORMAL 61424 M 205/ 177 PPM DISPOSITION 61433 M 2051177 PPM NORMAL DATE TIME GRADE OBSERVATIONS     ___ ._______________ 02 27 01 8: ll P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8~ 12 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9~ 20 P SCHEDULED EUTHANASIA 02 27 01 8~ 12 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8: 13 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8: 13 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8: 13 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8: 14 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 a: 14 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 a: 15 P No SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8: 15 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8: 16 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8: 16 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 28 P SCHEDULED EUTHANASIA 02 27 01 8: 17 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 29 P SCHEDULED EUTHANASIA 02 27 01 a: 17 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 27 01 9: 29 P SCHEDULED EUTHANASIA 02 27 01 8: 18 P NO SIGNIFICANT CLINICAL OBSERVATIONS ___________D E ______

GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PROJECT NO.: WIL 417001A SPONSOR: HAP TASK FORCE TABLE 14 (DETAILED PHYSICAL EXAMINATIONS/ DISPO. YITIONS) ACUTE INH. TOX (WITH HISTOPATHOLGGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 27 01 TO 02 27 01 PAGE 2 ________________________________________

ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS _______________________________________

61433 M 61418 M 61418 M 61418 M 61418 M 205/ 177 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 14741840 PPM 61418 M 1474/ 840 PPM 61423 M 1474/ 840 PPM 61423 M 1474/ 840 PPM 61423 M 1474/ 840 PPM 61423 M 1474/ 840 PPM 61423 M 3.4741840 PPM 61428 M 1474/ 840 PPM 61428 M 14741840 PPM 61428 M 1474/ 840 PPM 61428 M 1474f840 PPM 61428 M 14741840 PPM 61428 M 1474/ 840 PPM 61429 M 1474/ 840 PPM 61429 M 1474/ 840 PPM 61429 M 1474/ 840 PPM 61429 M 1474/ 840 PPM 61436 M 61436 M 1474/ 840 PPM 1474/ 840 PPM DISPOSITION DISPOSITION BEHAVIOR/ CNS BODY/ INTEGUMENT EYES/ EARS/ NOSE ORAL/ DENTAL DISPOSITION BEHAVIOR/ CNS EYES/ EARS/ NOSE EXCRETA ORAL/ DENTAL DISPOSITION BEHAVIOR/ CNS BODY/ INTEGUMENT CARDIO PULMONARY EYES/ EARS/ NOSE EXCRETA DISPOSITION BEHAVIOR/ CNS BODY/ INTEGUMENT EYES/ EARS/ NOSE DISPGSITION BODY/ INTEGUMENT 02 27 01 9: 29 P 02 27 01 9: 29 P 02 27 01 8: 20 P 02 27 01 8: lY P 02 27 01 8: 18 P 02 27 01 8: lY P 02 27 01 8: 20 P 02 27 01 8: 19 P 02 27 01 9: 29 P 02 27 01 8: 22 P 02 27 01 8: 21 P 02 27 01 8: 21 P 02 27 01 8~ 22 P 02 27 01 9: 29 P 02 27 01 8: 23 P 02 27 01 8~ 23 P 02 27 01 8: 23 P 02 27 01 8: 23 P 02 27 01 8: 24 P 02 27 01 9: 29 P 02 27 01 8: 26 P 02 27 01 8: 25 P 02 27 01 a: 25 P 02 27 01 a: 25 P 02 27 01 9: 29 P 02 7.7 01 0: 27 P SCHEDULED EUTHANASIA SCHEDULED EUTHANASIA LETHARGIC BODY COOL TO TOUCH DRIED RED MATERIAL AROUND NOSE CLEAR DISCHARGE RIGHT EYE CLEAR DISCHARGE LEFT EYE DRIED RED MATERIAL AROUND MOUTH SCHEDULED EUTHANASIA LETHARGIC DRIED RED MATERIAL AROUND NOSE URINE BROWNISH RED IN COLOR DRIED RED MATERIAL AROUND MOUTH SCHEDULED EUTHANASIA LETHARGIC BODY COOL TO TOUCH RESPIRATION RATE DECREASED DRIED RED MATERIAL AROUND NOSE URINE BROWNISH RED IN COLOR SCHEDULED EUTHANASIA LETHARGIC BODY COOL TO TOUCH CLEAR DISCHARGE LEFT EYE DRIED RED MATERIAL AROUND NOSE SCHEDULED EUTHANASIA BODY COOL TO TOUCH ___________________~ GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PROJECT NO.: WIL 417001A SPONSOR: HAP TASK FORCE _____ ANIMAL SEX GROUP 61450 F 0 PPM 61450 F 0 PPM 61464 F 0 PPM 61464 F 0 PPM 61469 F 0 PPM 61469 F 0 PPM 61490 F 0 PPM 61490 F 0 PPM 61494 F 0 PPM 61494 F 0 PPM 61454 F 60145 PPM 61454 F 60/ 45 PPM 61457 F 60145 PPM 61457 F 60/ 45 PPM 61466 F 60/ 45 PPM 61466 F 60/ 45 PPM 61487 F 60/ 45 PPM 61487 F 60/ 45 PPM 61492 F 60/ 45 PPM 61492 F 60/ 45 PPM 61448 F 2051177 PPM 61448 F 205/ 177 PPM 61463 F 205/ 177 PPM 61463 F 205/ 177 PPM 61468 F 205/ 177 PPM 61468 F 205/ 177 PPM 61483 F 205/ 177 PPM 61483 F 205/ 177 PPM 61489 F 205/ 177 PPM , TABLE 14 (DETAILED PHYSICAL EXAMINATI~ NS/ DISPO~ ITIONS) ACUTE INH. TOX (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 28 01 TO 02 28 01 PAGE 3 __ CATEGORY __ NORMAL DISPOSITION NORMAL DISWSITION NORM& L DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DISPOSITION NORMAL DATE TIME GRADE OBSERVATIONS ,_______________ 02 28 01 7: 29 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 30 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHT+ NIZATION 02 28 01 7: 30 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 30 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 31 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 31 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 31 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 32 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 32 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 33 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 33 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 34 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 08 P SCHEDULED EUTHANIZATION 02 28 01 7: 34 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 09 P SCHEDULED EUTHANIZATION 02 28 01 7: 35 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 28 01 lo: 09 P SCHEDULED EUTHANIZATION 02 28 01 7: 35 P NO SIGNIFICANT CLINICAL OBSERVATIONS ________________________________________"

GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PROJECT NO.: WIL 417001A SPONSOR: HAP TASK FORCE __________________________ ANIMAL SEX GROUP ___________________ 61489 61445 61445 61445 61460 61460 61460 61460 61460 61478 61478 63.478 61478 61478 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 61485 F 1474/ 840 PPM 61485 F 1474/ 840 PPM 205/ 177 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM __

 TABLE 14 (DETAILED PHYSICAL EXAMINATIONS/ DISPOS3TIONS) ACUTE INH. TOX (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN FATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 28 01 TO 02 28 01 PAGE 4 .__________________"

CATEGORY DATE TIME GRADE OBSERVATIONS .___________________~ DISPOSITION 02 28 01 lo: 09 P SCHEDULED EUTHANIZATION DISPOSITION 02 28 03. lo: 09 P SCHEDULED EUTHANIZATION BODY/ INTEGUMENT 02 28 01 7~ 38 P BODY COOL TO TOUCH EYES/ EARS/ NOSE 02 28 01 7: 39 P DRIED RED MATERIAL AROUND LEFT EYE 02 28 01 7: 40 P DRIED RED MATERIAL AROUND RIGHT EYE DISPOSITION 02 28 01 9: 32 P EUTHANIZED IN EXTREMIS BEHAVIOR/ CNS 02 28 01 7: 41 P LETHARGIC 02 28 01 7: 42 P ATAXIA BODY/ INTEGUMENT 02 28 01 7: 41 P BODY COOL TO TOUCH CARDIO PULMONARY 02 28 01 7: 41 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 28 01 7: 42 P CLEAR DISCHARGE RIGHT EYE DISPOSITION 02 28 01 8: 17 P FOUND DEAD BEHAVIOR/ CNS 02 28 01 7: 43 P PROSTRATE BODY/ INTEGUMENT 02 28 01 7: 44 P BODY COOL TO TOUCH CARDIO PULMONARY 02 28 01 7~ 44 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 28 01 7: 44 P CLEAR DISCHARGE RIGHT EYE 02 28 01 7: 44 P CLEAR DISCHARGE LEFT EYE NORMAL 02 28 01 7: 46 P NO SIGNIFICANT CLINICAL OBSERVATIONS DISPOSITION 02 28 01 lo: 09 P SCHEDULED EUTHANIZATION co ________________________________________ GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PCRDv4.01 06/ 12/ 2001 R: O6/ 12/ 2001 PAGE 1 PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 15 (IMMEDIATELY FOLLOWING EXPOSURE) ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 TO 02 25 01 ____________________

ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS __________________ 61409 M 0 PPM 61410 M 0 PPM 61419 M 0 PPM 61432 M 0 PPM 61434 M 0 PPM 61396 M 5% PPM 61405 M 58 PPM 61413 M 58 PPM 61416 M 58 PPM 61431 M 58 PPM 61399 M 181 PPM 61411 M 181 PPM 61441 M 181 PPM 61442 M 181 PPM 61443 M 181 PPM 61397 M 1527 PPM 61397 M 1527 PPM 61397 M 1527 PPM 61406 M 61406 M 61406 M 1527 PPM 1527 PPM 1527 PPM 61408 M 61408 M 61408 M 1527 PPM 1527 PPM 1527 PPM 61421 M 61421 M 1527 PPM 1527 PPM NORMAL 02 25 01 13: 29 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 29 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 29 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 30 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 30 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 25 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 25 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 26 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 26 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 27 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 14: 15 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 14: 15 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 14: 15 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 14: 16 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 14: 16 P NO SIGNIFICANT CLINICAL OBSERVATIONS BEHAVIOR/ CNS 02 25 01 13: 42 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 42 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 25 01 13: 43 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 43 P CLEAR DISCHARGE LEFT EYE BEHAVIOR/ CNS 02 25 01 13: 43 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 44 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 25 01 13: 44 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 44 P CLEAR DISCHARGE LEFT EYE BEHAVIOR/ CNS 02 25 01 13: 44 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 44 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 25 01 13: 44 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 45 P CLEAR DISCHARGE LEFT EYE BEHAVIOR/ CNS 02 25 01 13: 45 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 45 P RESPIRATION RATE DECREASED ____________________________________ GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PAGE 2 PROJECT NO.: WIL 417001 SPONSOR: HAP TASK FORCE TABLE 15 (IMMEDIATELY FOLLOWING EXPOSURE1 ACUTE INH. TOX. (WITH HISTOPATHOLOOY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 TO 02 25 01 ___________________~

ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS __________________ 61421 M 1527 PPM 61439 M 61439 M 61439 M 1527 PPM 1527 PPM 1527 PPM 61446 F 61452 F 61453 F 61456 F 61491 F 61467 F 61470 F 61472 F 61475 F 61417 F 61449 F 61474 F 61482 F 61486 F 61493 F 61451 F 61451 F 61451 F 0 PPM 0 PPM 0 PPM 0 PPM 0 PPM 58 PPM 50 PPM 58 PPM 58 PPM 58 PPM 181 PPM 181 PPM 181 PPM 181 PPM 181 PPM 1527 PPM 3.527 PPM 1527 PPM 61462 F 61462 F 61462 F 1527 PPM 1527 PPM 1527 PPM EYES/ EARS/ NOSE 02 25 01 13: 45 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 45 P CLEAR DISCHARGE LEFT EYE BEHAVIOR/ CNS 02 25 01 13: 45 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 46 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 25 01 13: 46 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13~ 46 P CLEAR DISCHARGE LEFT EYE NORMAL 02 25 01 13: 30 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 31 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 31 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 31 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 31 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAG 02 25 01 13: 27 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 28 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 28 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 28 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 13: 29 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 14: 16 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORMAL 02 25 01 14: 17 P NO SIGNIFICANT CLINICAL OBSERVATIONS NORM& L 02 25 01 14: 17 P NO SIGNIFICANT CLINICAL OBSERVATIONS EYES/ EARS/ NOSE 02 25 01 14: 18 P DRIED RED MATERIAL AROUND RIGHT EYE NORMAL 02 25 01 14: 18 P NO SIGNIFICANT CLINICAL OBSERVATIONS BEHAVIOR/ CNS 02 25 01 13: 47 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 47 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 25 01 13: 47 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 47 P CLEAR DISCHARGE LEFT EYE BEHAVIOR/ CNS 02 25 01 13: 47 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 48 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 25 01 13: 48 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 48 P CLEAR DISCHARGE LEFT EYE _______________________________________ GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PROJECT NO.: NIL 417001 SPONS0R: HA. P TASK FORCE TABLE 15 (IMMEDIATELY FOLLOWING EXPOSURE) ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 TO 02 25 01 PAGE 3 ____________"_______~

ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS ____________________ 61476 F 1527 PPM 61476 F 1527 PPM 61476 F 1527 PPM 61481 F 1527 PPM 61481 F 1527 PPM 61481 F 1527 PPM 61484 F 1527 PPM 61484 F 1527 PPM 61484 F 1527 PPM ____ GRADE CODE: 1  SLIGHT 2 r _ BEHAVIOR/ CNS 02 25 01 13: 49 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 49 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 25 01 13: 49 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 49 P CLEAR DISCHARGE LEFT EYE BEHAVIOR/ CNS 02 25 01 13: 49 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 49 P RESPIRATION RATE DECREASED EYES/ EARS/ NOSE 02 25 01 13: 49 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 49 P CLEAR DISCHARGE LEFT EYE BEHAVIOR/ CNS 02 25 01 13: 49 P ANIMAL SLEEPING CARDIO PULMONARY 02 25 01 13: 50 P RESPIRATION RATE DECREASED EYES/ BARS/ NOSE 02 25 01 13: 50 P CLEAR DISCHARGE RIGHT EYE 02 25 01 13: 50 P CLEAR DISCHARGE LEFT EYE MODERATE 3  SEVERE P  PRESENT PCRDv4.04 10/ 29/ 2001 PROJECT NO.: WIL 417001A' SPONSOR: HAP TASK FORCE TABLE 15 (IMMEDIATELY FOLLOWING EXPOSURE) ACUTE INH. TOX (WITH HISTOPATHOLGGY~ STUDY OF 1.1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 26 01 TO 02 26 01 PAGE I ______________________________ I_______~~

ANIMAL SEX GROUP CATEGORY DATE TIME GRADE OBSERVATIONS ________________________________________~~ 61400 M 0 PPM 61403 M 0 PPM 61404 M 0 PPM 61414 M 0 PPM 61420 M 0 PPM 61395 M 60/ 45 PPM 61398 M 60/ 45 PPM 61425 M 60/ 45 PPM 61430 M 60/ 45 PPM 61435 M 60/ 45 PPM 61407 M 205/ 177 PPM 61415 M 205/ 177 PPM 61422 M 2051177 PPM 61424 M 205/ 171 PPM 61433 M 2051177 PPM 61418 M 14741840 PPM 61423 M 1474/ 840 PPM 61423 M 61428 M 61428 M 61429 M 61429 M 61436 M 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 1474/ 840 PPM 14741840 PPM 14741840 PPM 61436 M 1474/ 840 PPM NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL NORMAL BEHAVIOR/ CNS BEHAVIOR/ CNS EYES/ EARS/ NOSE BEHAVIOR/ CNS CARDIO PULMONARY REHAVIQR/ CNS EYES/ EARS/ NOSE BEUAVIOR/ CNS EYES/ EARS/ NOSE 02 26 01 11: 05 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 06 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 07 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 08 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 08 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 26 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 26 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 27 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 27 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 28 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 41 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 41 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 42 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 42 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 43 P NO SIGNIFICANT CLINICAL OBSERVATIONS 02 26 01 11: 50 P LETHARGIC 02 26 01 11: 52 P LETHARGIC 02 26 01 11~ 53 P ATAXIA 02 26 01 11: 54 P CLEAR DISCHARGE RIGHT EYE 02 26 01 11: 55 P ANIMAL SLEEPING 02 26 01 11: 56 P RESPIRATION RATE DECREASED 02 26 01 11: 56 p ANIMAL SLEEPING 02 26 01 11: 57 P CLEAR DISCHARGE LEFT EYE 02 26 01 11: 58 P LETHARGIC 02 26 01 11: 58 P ATAXIA 02 26 01 11: 58 P CLEAR DISCHARGE RIGHT EYE 02 26 01 11: 58 P CLEAR DISCHARGE LEFT EYE ________________________________________ GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT PROJECT NO.: WIL 417001A SPONSOR: HAP TASK FORCE TABLE 15 (IMMEDIATELY FOLLOWING EXPOSURE) ACUTE INH. TOX (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 27 01 TO 02 27 01 PAGE 2 ________"______________________________

ANIMAL SEX GROUP CATEG0RY DATE TIME GRADE OBSERVATIONS __________________""

61450 F 0 PPM 61464 F 0 PPM 61469 F 0 PPM 61490 F 0 PPM 61494 F 0 PPM 61454 F 60/ 45 PPM 61457 F 60/ 45 PPM 61466 F 60145 PPM 61487 F 60/ 45 PPM 61492 F 60/ 45 PPM 61448 F 2051177 PPM 61463 F 205/ 177 PPM 61468 F 205/ 177 PPM 61483 F 2051177 PPM 61489 F 2051177 PPM 61445 F 1474/ 840 PPM 61445 F 1474/ 840 PPM 61445 F 14741840 PPM 61460 F 61460 F 61460 F 1474/ 840 PPM 14741840 PPM 1474/ 840 PPM 61478 F 61478 F 61478 F 14741840 PPM 1474/ 840 PPM 14741040 PPM 61485 F 61485 F 1414/ 040 PPM 14741840 PPM NORMAL 02 27 01 12: 25 P NORMAL 02 27 01 12: 25 P NORMAL 02 27 01 12: 26 P NORMAL 02 27 01 12: 26 P NORMAL 02 27 01 12: 26 P NORMAL 02 27 01 12: 32 P NORMAL 02 27 01 12: 32 P NORMAL . 02 27 01 12: 32 P NORMAL 02 27 01 12: 33 P NORMAL 02 27 01 12: 33 P NORMAL 02 27 01 12: 35 P NORMAL 02 27 01 12: 36 P NORMAL 02 27 01 12: 36 P NORMAL 02 27 01 12: 37 P NORMAL 02 27 01 12: 37 P BEHAVIOR/ CNS 02 27 01 13: 02 P CARDIO PULMONARY 02 27 01 13: 02 P EYES/ EARS/ NOSE 02 27 01 13: 03 P 02 27 01 13: 03 P BEHAVIORfCNS 02 27 01 13: 09 P CARDIO PULMONARY 02 27 01 13: OP P EYES/ EARS/ NOSE 02 27 01 13: l. O P 02 27 01 13: lO P BEHAVIOR/ CNS 02 27 01 13: lO P CARDIO PULMONARY 02 27 01 13: lO P EYES/ EARS/ NOSE 02 27 01 13: lO P 02 27 01 13: lO P BEHAVIOR/ CNS 02 27 01 13: ll P CARDIO PULMONARY 02 27 01 13: ll P NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS No SIwIFImiT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS NO SIGNIFICANT CLINICAL OBSERVATIONS ANIMAL SLEEPING RESPIRATION RATE DECREASED CLEAR DISCHARGE RIGHT EYE CLEAR DISCHARGE LEFT EYE ANIMAL SLEEPING RESPIRATION RATE DECREASED CLEAR DISCHARGE RIGHT EYE CLEAR DISCHARGE LEFT EYE ANIMAL SLEEPING RESPIRATION RATE DECREASED CLEAR DISCHARGE RIGHT EYE CLEAR DISCHARGE LEFT EYE ANIMAL SLEEPING RESPIRATION RATE DECREASED ________________________________________"

GRADE CODE: 1  SLIGHT 2  MODERATE 3  SEVERE P  PRESENT 115 PROJECT NO.: WIL 417001 SPONS0R: HA. P TASK FORCE TABLE 16 (RESPONSE TO NOVEL STIMULUS) ACUTE INH. TOX. (WITH HIST~ PATHOLOGY) STUDY OF l, l, z~ C~ IN RATS INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 PAGE 1 ANIMAL SEX GROUP DATE TIME ____________________

61409 M 61410 M 61419 M 61432 M 61434 M 61396 M 61405 M 61413 M 61416 M 61431 M 61399 M 61411 M 61441 M 61442 M 61443 M 61397 M 61406 M 61408 M 61421 M 61439 M 0 PPM 02 25 01 14: 15 0 PPM 02 25 01 14: 15 0 PPM 02 25 01 14: 15 0 PPM 02 25 01 14: 15 0 PPM 02 25 01 14: 15 58 PPM 02 25 01 14: 18 58 PPM 02 25 01 14: 18 58 PPM 02 25 01 14: lE 58 PPM 02 25 01 14: 18 50 PPM 02 25 01 14: 18 181 PPM 02 25 01 14: 22 181 PPM 02 25 01 14: 22 181 PPM 02 25 01 14: 22 181 PPM 02 25 01 14: 22 181 PPM 02 25 01 14: 22 1527 PPM 02 25 01 14: 25 1527 PPM 02 25 01 14: 25 1527 PPM 02 25 01 14: 25 1527 PPM 02 25 01 14: 25 1527 PPM 02 25 01 14: 25 OBSERVATIONS ________________ NOTABLY INCREASED REACTION NO REACTION TO STIMULUS NOTABLY INCREASED REACTION NOT VISIBLE NO REACTION TO STIMULUS NOTABLY INCREASED REACTION SLIGHT REACTION NOTABLY INCREASED REACTION NOT VISIBLE NO REACTION TO STIMULUS NO REACTION TO STIMULUS SLIGHT REACTION SLIGHT REACTION NOT VISIBLE NO REACTION TO STIMULUS NO REACTION TO STIMULUS No REACTION TO STIMULUS NO REACTION TO STIMULUS NOT VISIBLE NO REACTION TO STIMULUS TABLE 16 (RESPONSE ~0 NOVEL STIMULUS) PROJECT NO.: WIL 417001 ACUTE INH. TOX. (WITH HISTOPATHOLOGY) STUDY OF 1,1,2 TCE IN RATS PAGE 2 SPONSOR: HAP TASK FORCE INDIVIDUAL CLINICAL OBSERVATIONS TABLE RANGE: 02 25 01 _____________"__________________________"

ANIMAL SEX GROUP DATE TIME OBSERVATIONS ________________________________________~"~~

61446 F 0 PPM 02 25 01 14: 15 NO REACTION TO STXMULUS 61452 F 0 PPM 02 25 01 14: lS NOTABLY INCREASED REACTION 61453 F 0 PPM 02 25 01 14: 15 NO REACTION TO STIMULUS 61456 F 0 PPM 02 25 01 14: 15 NO REACTION TO STIMULUS 61491 F 0 PPM 02 25 01 14: 15 NOT VISIBLE 61467 F 58 PPM 02 25 01 14: 18 NO REACTION TO STIMULUS 61470 F 58 PPM 02 25 01 14: 18 NOTABLY INCREASED REACTION 61472 F 58 PPM 02 25 01 14: 18 NO REACTION TO STIMULUS 61475 F 58 PPM 02 25 01 14: 18 NOTABLY INCREASED REACTION 61477 F 58 PPM 02 25 01 14: 18 NOT VISIBLE 61449 F 181 PPM 02 25 01 14: 22 NO REACTION TO STIMULUS 61474 F 181 PPM 02 25 01 14: 22 SLIGHT REACTION 61482 F 181 PPM 02 25 01 14: 22 SLIGHT REACTION 61486 F 181 PPM 02 25 01 14: 22 NO REACTION TO STIMULUS 61493 F 181 PPM 02 25 01 14: 22 NOT VISIBLE 61451 F 1527 PPM 02 25 01 14: 25 NO REACTION TO STIMULUS 61462 F 1527 PPM 02 25 01 14: 25 NO REACTION TO STIMULUS 61476 F 1527 PPM 02 25 01 14: 25 NO REACTION TO STIMULUS 61481 F 1527 PPM 02 25 01 14~ 25 NO REACTION TO STIMULUS 61484 F 1527 PPM 02 25 01 14: 25 NOT VISIBLE ________________________________________