United States
Environmental Protection
Agency
Health Effects Research
Laboratory
Cincinnati OH 45268
EPA-600/1-80-010
January 1980
Research and Development
vvEPA
Teratologic
Effects of Long-Term
Exposure to Diesel
Exhaust Emissions
(Rats)
-------
RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4 Environmental Monitoring
5. Socioeconomic Environmental Studies
6. Scientific and Technical Assessment Reports (STAR)
7 Interagency Energy-Environment Research and Development
8. "Special" Reports
9. Miscellaneous Reports
This report has been assigned to the ENVIRONMENTAL HEALTH EFFECTS RE-
SEARCH series. This series describes projects and studies relating to the toler-
ances of man for unhealthful substances or conditions. This work is generally
assessed from a medical viewpoint, including physiological or psychological
studies. In addition to toxicology and other medical specialities, study areas in-
clude biomedical instrumentation and health research techniques utilizing ani-
mals — but always with intended application to human health measures.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
-------
EPA-600/1-80-010
January 1980
TERATOLOGIC EFFECTS OF LONG-TERM
EXPOSURE TO DIESEL EXHAUST EMISSIONS (RATS)
by
Kathleen M. Werchowski, Vincent W. Chaffee,
and G. Bruce Briggs
WIL Research Laboratories, Inc.
3154 Exon Avenue
Cincinnati, Ohio 45241
Contract No. 68-03-2652
Project Officer
WiUiam E. Pepelko
Laboratory Studies Division
Health Effects Research Laboratory
Cincinnati, Ohio 45268
HEALTH EFFECTS RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U. S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO 45268
-------
DISCLAIMER
This report has been reviewed by the Health Effects Research Laboratory,
U.S. Environmental Protection Agency, and approved for publication. Approval
does not signify that the contents necessarily reflect the views and policies
of the U.S. Environmental Protection Agency, nor does mention of trade names
or commercial products constitute endorsement or recommendation for use.
11
-------
FOREWORD
The use of diesel engines for both personal and commercial transportation
is increasing, and there is very little information presently available
concerning the toxicology of whole emissions from diesel exhaust.
This study was performed by MIL Research Laboratories, Inc. to contribute
information as to the potential teratogenic effects of diesel exhausts
when rats were exposed to diesel exhaust emissions. The procedures
utilized in exposure and fetal evaluations were developed by the Food
and Drug Administration for evaluating teratogenic effects of new drugs.
R". J. Garner
Director
Health Effects Research Laboratory
111
-------
ABSTRACT
This research project was initiated with the objective of evaluating the potential
for diesel exhaust emissions to produce malformations in rat fetuses. The dams were
exposed by the inhalation route to a 10% concentration of diesel exhaust emissions in
inhalation chambers on days 6 through 15 of gestation. Methods used conform to the
guideline developed by the Food and Drug Administration for evaluating teratogenic
effects in rats.
The results of the exposure of pregnant rats indicate that diesel exhaust
emissions have no effect upon the normal development of rat fetuses.
Diesel exhaust emissions did not effect any of the parameters evaluated to
assess maternal toxicity nor total number of fetuses born.
This report was submitted in fulfiUment of Contract No. 68-03-2652 by WIL
Research Laboratories, Inc. under the sponsorship of the U. S. Environmental Protec-
tion Agency. This contract includes the evaluation of the teratogenic effect of diesel
exhaust emissions in rabbits and this work is continuing and will be reported separately
from this report. This report covers'the period September 15, 1978 to February 7,
1979 when the work was completed.
IV
-------
CONTENTS
Foreword iii
Abstract iv
Tables vi
Abbreviations vii
1 Introduction 1
2 Experimental Design 2
Test Material 2
Treatment Preparation and Administration 2
Animals - Requisition and Care 2
Dosage Selection 3
Mating Procedure 3
Treatment and Dosing Schedule 3
Observations During Gestation Period 3
Observations at Time of Laparotomy 3
Fetal Examination 4
3 Results 5
General Observations 5
Body Weights of Adult Male and Female Rats 5
Fertility - Mating Data 5
Laparotomy Data 12
Fetal Visceral Data 12
Fetal Skeletal Data 12
Gross Necropsies 13
-------
TABLES
Number Page
1 Individual Body Weights for Darns Which Were Exposed to Clean Air
(Group 1) 6
2 Individual Body Weights for Dams Which Were Exposed to Diesel
Exhaust Inhalation (Group 2) 7
3 Individual Weekly Body Weights for Male Rats Which Mated with Group
1 Females 8
4 Individual Weekly Body Weights for Male Rats Which Mated with Group
2 Females 9
5 Individual Mating Data 10
6 Gross Fetal Examination at Time of Laparotomy 14
7 Individual Fetal Data for Dams Which Were Exposed to Clean Air; Sex
Distribution, Mean Body Weight, Individual Fetus Weight, Group Fetal
Weight, Fetal Position in Uterine Horn 18
8 Individual Fetal Data for Dams Which Were Exposed to Diesel Exhaust
Emissions; Sex Distribution, Mean Body Weight, Individual Fetus
Weight, Group Fetal Weight, Fetal Position in Uterine Horn 19
9 Individual Fetal Data for Dams Which were Exposed to Clean Air;
Number of Fetuses Viable and Dead, Early Resorptions, Implantation
Sites and Corpora Lutea 20
10 Individual Fetal Data for Dams Which Were Exposed to Diesel Exhaust
Emissions; Number of Fetuses Viable and Dead, Early Resorptions,
Implantation Sites and Corpora Lutea 21
11 Soft Tissue Examination of the Fetuses 22
12 Skeletal Examination of Term Fetuses - Mean Number of Ossified
Elements (Group 1) 23
13 Skeletal Examination of Term Fetuses - Mean Number of Ossified
Elements (Group 2) 25
14 Skeletal Examination of the Fetuses 28
15 Gross Necropsy Results for 20-Day Laparotomy Female Rats 30
vi
-------
ABBREVIATIONS
P = Pregnant
NP = Not Pregnant
NN = Not Necessary
BB = Did Not Breed
C - Cesarean Section
S = Sacrificed
B = Bred
M = Male
F := Female
L = Left
R = Right
vii
-------
SECTION 1
INTRODUCTION
Diesel fuel has increased in the use of diesel engines in vehicular traffic and there is
little information presently available concerning the toxicology of whole emissions from
diesel exhaust, thus the proposal made in RFP No. Cl 77-0221 was performed on
laboratory pregnant animals to establish potential teratogenic effects during the period
of fetal development.
The objective of this study was to evaluate the potential of diesel exhaust emissions to
produce malformations and/or other teratogenic effects in the unborn fetus when dams
were exposed by means of inhalation during the period of organogenesis. The methods
described herein conform to the guideline developed by FDA in 1966 .
Diesel exhaust emissions produced no malformations nor other teratogenic effects in the
unborn rat fetus when dams were exposed to a 10% concentration of diesel exhaust by
means of inhalation during the period of organogenesis.
'''Guidelines for Reproductive Studies for Safety Evaluation of Drugs for Human Use,
January 1966, FDA.
-------
SECTION 2
EXPERIMENTAL DESIGN
TEST MATERIAL
The test material used in this study was diesel exhaust emissions generated by a diesel
engine located at the EPA Center Hill Laboratory, Cincinnati, Ohio. As a control, clean
air was administered by the same means; at chambers also located at EPA Center Hill
Laboratory. The first day of inhalation exposure was November 1, 1978.
TREATMENT PREPARATION AND ADMINISTRATION
A ratio of ten percent diesel exhaust emission to ninety percent clean air was
administered to the test group (Group 2). Inhalation was the route of administration in
chambers 4 1/2 feet square. The control group (Group 1) was also exposed in 4 1/2 feet
square inhalation chambers with 100% clean air. The Center Hill EPA Laboratory was
responsible for maintaining the proper exposure rates, monitoring procedures, and care
of the laboratory animals during the exposure period.
ANIMALS - REQUISITION AND CARE
Forty female albino rats and twenty male albino rats, Sprague Dawley strain, ten weeks
of age, were obtained from Charles River Breeding Labs, Portage, Michigan. The rats
were received on September 15, 1978.
The animal care procedures for the female rats were the responsibility of WIL Research
Laboratories, Inc., during the quarantine period and Days 0 through 5, and 16 through 20
of gestation. The EPA Center Hill Laboratory, Cincinnati, Ohio, was responsible for the
animal care of the female animals during the exposure period which was Day 6 through
15 of gestation.
The animal care for the male rats was the responsibility of WIL Research Laboratories,
Inc., for the entire duration of the study. All animal care procedures used in the
performance of this study adhered to the principles defined in the Guide for Laboratory
Animal Facilities and Care and The Animal Welfare Act and Amendments.
All animals at WIL were kept in a conventional room in AALAC approved cages during
the quarantine and the nonexposure periods. Purina Laboratory Chow and fresh tap
water were available _ad libitum during the quarantine period and the nonexposure
periods.
All rats were examined prior to selection for this study and only those animals
determined to be in good health were selected for testing. During the quarantine period
the animals were permanently identified by the ear-punch method.
-------
Following the exposure time in the chambers at the EPA facilities, the rats remained in
individual cages at which time clean air was administered. The animals were maintained
in the individual cages within the chamber rooms which used a 12-hour light cycle during
the animals' daily routines. The inhalation chamber was thoroughly sanitized between
each exposure period to minimize the possibility of disease transmission. At this time,
individual cages were placed outside the chambers and then replaced after cleaning.
DOSAGE SELECTION
The EPA conducted preliminary studies to determine the 10% concentration of diesel
exhaust emissions in the exposure chambers. The Environmental Protection Agency
personnel were responsible for assuring chamber concentrations and exposure time.
MATING PROCEDURE
The female animals were randomly selected and placed in either the test group or
control group. Each of the twenty male rats was mated to two females. The first of the
two females was housed with the male until a copulatory plug was observed. After
evidence of copulation, the second female was housed with the male and observed for a
copulatory plug.
If a plug, or other evidence of copulation, was not observed in a female after 14 days of
mating, the female was mated with another male. The day a copulatory plug was
observed was considered to be Day 0 of gestation.
TREATMENT AND DOSING SCHEDULE
The female rats were exposed on Days 6 through 15 of gestation for 8 hours per day.
Group 1 was exposed to 100% clean air while Group 2 was exposed to the 10% diesel
exhaust emissions and 90% clean air.
OBSERVATIONS DURING GESTATION PERIOD
All female rats were examined daily for signs of toxicity, general health and behavior.
Each dam was weighed on Days 0, 6 through 15, and 20 of gestation.
One female in the control group (1207-17) was inadvertently killed on Day 18 of her
pregnancy due to technicians error as she thought the rat had not been successfully bred.
The rat was not exposed to the clean air.
All males were observed daily from the initiation of mating until the cesarean sections
were completed, then they were kiUed.
OBSERVATIONS AT TIME OF LAPAROTOMY
The female rats were terminated on Day 20 of gestation with an intracardial dose of
sodium pentobarbital. A cesarean section and a gross necropsy of each female was
performed. The following observations were made:
1. Total number of fetuses.
2. Total number of live and dead fetuses.
3. Number of resorbed or resorbing fetuses.
-------
4. Number of implant sites.
5. Number of corpora lutea.
6. Distribution of fetuses within the uterine horns.
7. Any remarkable gross pathology in the dam.
8. Examination of the fetuses as removed from the uterus:
a. External conformation and anomalies.
b. Individual body weight of each fetus and total litter weight.
c. Sex of each fetus.
The last cesarean section was performed on December 6, 1978.
FETAL EXAMINATION
Each fetus was examined by a teratologist for external anomalies of the head, mouth,
dorsal, and ventral aspects of the body, limbs, and tail. One-third of the fetuses were
placed in Bouin's fixative and examined for gross internal anomalies by careful Wilson
Sectioning . The remaining two-thirds of the fetuses were placed in 95% ethyl alcohol
and examined for skeletal anomalies following preparation of the skeletons by the
Alizarin Red S Procedure . Fetal examinations were completed on February 7, 1979.
"Environment and Birth Defects," James G. Wilson, Children's Hospital Research
Foundation, University of Cincinnati College of Medicine, 1973.
3
"Pathology and Toxicology, Technique Manual," edited by L. Z. Saunders; Research and
Development Division, Smith, Kline and French Laboratories, Philadelphia, Pennsyl-
vania, 1969, pages 1-13.
-------
SECTION 3
RESULTS
GENERAL OBSERVATIONS
Female rats were not observed to have any change in behavior nor signs of toxicity
during the study. Dried red material was observed around the nose of one control rat on
Day 15 and at the perimeter of the eyes of one rat on Days 18 through 21 of the study.
Six rats exposed to diesel exhaust were observed to be hyperactive during the first week
of exposure and head swaying was observed in four rats on one day of the exposure
period. The reason for these observations is not clearly understood, but probably is due
to irritation of the mucous membranes caused by the exhaust emissions. There was black
particulate matter on the chamber, cages and rats in the diesel exhaust chambers on the
days when these signs were observed and possibly contributed to irritation. The male
rats were not exposed to the diesel exhaust emissions and remained within the limits of
normal during the study.
BODY WEIGHTS OF ADULT MALE AND FEMALE RATS
Diesel exhaust emissions produced no reduction in body weight gains of females exposed
during Days 6 through 15 following the observation of a copulatory plug. The three
females found to be not pregnant actually lost weight during the study and the pregnant
rats in both the control and exposure groups gained weight at a similar rate.
The male rats were not exposed to diesel exhaust but they were weighed during the time
that the females were exposed and these animals gained weight at a normal rate (Tables
1, 2, 3 and 4).
FERTILITY - MATING DATA
Individual mating data for females in Groups 1 and 2 are presented in Table 5. The
fertility index for the females for each group was calculated using the following formula:
_ ..... T , No. Pregnant Females „ 1nn
Fertility Index = No. Females Mated X 10°
Group 1 females exposed to clean air had a conception rate of 89% (17 of 19 animals
were pregnant), and the conception rate of females exposed to diesel exhaust was 95%
(19 of 20 animals were pregnant) (Table 5).
-------
TABLE 1
INDIVIDUAL BODY WEIGHTS FOR DAMS WHICH WERE
EXPOSED TO CLEAN AIR (GROUP I - CONTROL)
Rat
Number
1207-2
1207-3
1207-5
1207-6
1207-8
1207-9
1207-11
1207-12
1207-14
1207-15
1207-171
01 1207-18
1207-20
1207-21
1207-23
1207-24
1207-26
1207-27
1207-29
1207-30
Day
0
260.6
274.6
255.3
274.4
325.0
379.6
308.0
288.9
292.2
258.8
-
308.5
260.2
288.5
305.7
276.1
270.8
283.9
285.1
283.4
Day
6
270.5
286.2
271.6
277.6
334.0
361.2
297.8
294.2
281.3
267.2
-
309.1
240.1
310.9
296.4
285.2
277.1
267.3
275.4
284.7
Day
7
269.8
291.8
274.2
274.3
336.0
369.5
309.4
308.3
300.1
276.4
-
317.8
256.2
314.7
298.4
296.8
273.2
277.5
283.6
286.5
Day
8
279.7
290.7
269.8
284.3
345.1
370.4
302.7
303.9
300.5
285.1
-
316.9
267.4
319.8
311.0
300.5
278.9
280.2
293.9
286.3
Day
9
278.6
294.5
268.6
275.0
346.5
370.3
311.2
310.3
307.7
268.7
-
314.5
263.1
322.0
321.0
308.7
283.1
271.9
302.8
291.8
Day
10
290.6
291.4
280.2
291.6
351.6
375.0
315.8
316.3
316.3
271.2
-
319.4
269.3
336.3
326.2
310.4
276.1
266.9
308.7
295.8
Day
11
292.7
293.6
281.1
302.1
359.8
379.9
324.4
316.5
319.7
285.7
-
330.0
272.5
340.0
338.4
310.1
290.6
270.7
303.9
301.7
Day
12
295.3
304.5
290.1
306.8
369.9
382.8
329.2
318.2
326.6
283.5
-
332.9
282.5
350.1
342.1
316.7
293.1
291.6
323.8
306.6
Day
13
299.6
303.9
297.4
306.5
368.0
390.4
336.8
320.1
332.7
297.0
-
345.2
281.5
350.6
346.3
323.4
299.1
287.2
330.2
315.1
Day
14
300.6
302.0
299.4
309.8
368.0
390.9
343.3
322.1
337.5
297.3
-
345.1
284.0
353.6
352.4
324.9
308.1
283.7
332.4
308.8
Day
15
.310.0
300.2
306.5
315.5
375.4
399.6
355.2
326.4
348.2
305.3
-
353.1
295.4
358.9
355.4
333.1
318.8
280.3
345.6
319.9
Day
20
398.9
304.8
349.3
397.4
473.1
463.7
417.5
399.1
423.4
363.8
-
434.3
367.5
435.5
441.6
387.3
366.6
278.2
433.1
369.6
% Body
Weight
1 ncrease
53
11
37
45
46
22
36
38
45
41
_
41
41
51
44
40
35
-2
52
30
P*
NP**
P
NP
P
P
P
P
P
P
P
P
P
F
F
F
F
P
P
NP
F
F
Average 288.4 288.8 295.5 299.3 300.5
* Pregnant
** Not Pregnant
1 Female was sacrificed due to miscalculation of pregnancy and non-exposure
311.2
318.2
322.7
324.4
331.7
395.0
37.2
-------
TABLE 2
INDIVIDUAL BODY WEIGHTS FOR DAMS WHICH WERE
EXPOSED TO DIESEL EXHAUST INHALATION (GROUP II - TEST)
Rat DaX
Number 0
1207-32 302.4
1207-33 252.3
1207-35 238,5
1207-36 303.1
1207-38 319.0
1207-39 308.8
1207-41 270.6
1207-42 280.2
1207-44 273.1
1207-45 299.6
^ 1207-47 315.7
1207-48 284.3
1207-50 285.3
1207-51 330.9
1207-53 284.5
1207-54 303.7
1207-56 310.3
1207-57 281.1
1207-59 277.8
1207-60 314.6
Average 291.8
* Pregnant
** Not Pregnant
Day
6
298.9
266.1
242.8
334.1
311.9
317.4
270.3
292.1
287.0
308.1
320.4
293.2
259.3
341.2
296.3
317.5
319.7
298.0
286.1
330.3
299.5
Day
7
308.3
269.8
254.6
329.2
332.8
325.2
278.8
286.4
287.8
309.4
316.9
296.5
267.3
344.4
307.0
317.6
325.4
300.4
286.6
330.1
303.7
Day
8
305.3
279.0
256.8
337.3
335.7
324.3
277.7
293.0
289.5
306.6
327.7
300.8
272.5
349.8
311.5
318.4
325.4
303.6
285.1
332.1
306.6
Day
9
309.5
280.4
256.5
339.3
341.6
327.4
280.2
300.8
295.3
311.8
329.0
295.8
271.5
350.8
315.7
325.6
331.3
305.4
290.8
332.3
309.6
Day
10
317.5
270.0
263.3
347.1
339.7
326.2
289.8
306.8
298.7
324.8
336.0
316.2
271.3
358.7
317.3
324.5
332.2
308.9
295.6
341.9
314.0
Day
11
323.1
254.3
272.3
351.5
342.4
336.5
290.7
310.6
306.6
326.5
348.1
305.0
269.5
368.1
320.0
331.9
333.9
314.7
296.9
329.9
316.6
Day
12
330.1
280.7
273.5
351.7
351.8
336.7
297.0
316.5
308.5
336.1
348.6
319.6
270.0
365.2
324.4
329.4
347.9
315.9
300.3
348.4
322.6
Day
13
329.1
289.4
253.7
360.2
359.9
345.6
303.0
324.6
309.6
333.8
346.5
316.3
271.1
370.9
331.0
338.6
357.1
328.6
311.8
349.3
326.5
Day
14
337.1
291.3
281.3
364.0
356.0
345.0
311.4
327.6
311.9
345.2
349.0
320.5
272.1
370.2
341.3
345.1
368.4
324.7
313.8
350.6
331.3
Day
15
347.4
294.8
283.8
376.6
368.1
358.4
318.3
338.9
320.4
361.1
358.3
335.4
273.0
379.5
318.2
356.1
379.6
336.5
323.9
365.6
339.7
Day
20
412.4
371.0
353.4
455.5
468.0
440.8
373.6
417.0
390.4
425.4
449.1
397.6
278.0
442.7
412.1
465.5
474.4
420.6
398.9
442.1
414.4
% Body
Weight
Increase
36
47
48
50
47
43
38
49
43
42
42
40
-3
34
45
53
53
50
44
41
42
P*
NP**
P
P
P
P
F
F
F
F
F
F
P
P
NP
P
P
P
P
P
P
P
-------
TABLE 3
WEEKLY BODY WEIGHTS FOR MALE RATS WHO MATED WITH
FEMALE RATS DOSED WITH 100% CLEAN AIR
GROUP I - CONTROL
Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7
Rat No.
1207-1
1207-4
1207-7
1207-10
1207-13
1207-16
1207-19
1207-22
1207-25
1207-28
Wt.(g)
421.9
394.7
405.6
381.7
436.6
401.0
446.4
431.8
428.1
404.1
Wt.(g)
422.6
395.6
410.1
397.7
454.8
416.0
459.9
450.6
438.8
405.8
Wt.(g)
460.6
425.5
445.8
424.3
488.8
443.9
489.5
479.1
473.9
428.1
Wt.(g)
474.4
458.1
453.3
451.3
515.7
459.4
513.7
516.2
484.9
440.1
Wt.(g)
479.3
451.4
460.1
452.9
521.8
472.3
514.3
537.0
498.1
453.3
Wt.(g)
491.6
467.3
471.5
471.9
527.4
482.6
523.3
545.2
503.6
463.7
Wt.(g)
511.4
507.8
489.1
500.1
555.7
502.6
543.9
572.5
532.9
477.3
Average 415.2 425.2 456.0 476.7 484.1 494.8 519.3
-------
TABLE 4
WEEKLY BODY WEIGHTS FOR MALE RATS WHO MATED WITH FEMALE RATS
DOSED WITH A CONCENTRATION OF 10% DIESEL EXHAUST EMISSION
GROUP II - TEST
Rat No.
1207-31
1207-34
1207-37
1207-40
1207-43
1207-46
1207-49
1207-52
1207-55
1 207-58
Average
Week 1
Wt.(g)
401.1
392.3
464.0
448.7
405.5
457.4
398.7
413.5
421.5
511.2
431.4
Week 2
Wt.(g)
410.6
407.2
476.1
463.8
431.1
466.5
412.9
412.5
408.3
513.1
440.2
Week 3
Wt.(g)
428.5
441.6
507.1
491.8
455.0
494.6
435.5
438.2
445.1
558.7
469.6
Week 4
Wt.(g)
460.0
455.8
537.5
523.9
471.5
516.9
447.8
444.7
466.2
595.5
492.0
Week 5
Wt.(g)
465.4
459.4
564.2
537.6
484.7
539.9
454.9
456.6
470.6
619.3
505.3
Week 6
Wt.(g)
465.7
466.6
578.5
544.0
486.1
558.2
463.6
468.1
495.2
624.4
515.0
Week 7
Wt.(g)
496.0
486.5
606.8
574.1
511.9
607.9
495.7
496.5
527.3
669.2
547.2
-------
Key
NN = Not Necessary
BB = Did Not Breed
C = C-Sectioned
S = Sacrificed
NP = Not Pregnant
B = Bred
TABLE 5
INDIVIDUAL MATING DATA
Clean Air Inhalation (Group 1)
Dam
Number
1207-2
1207-3
1207-5
1207-6
1207-8
1207-9
1207-11
1207-12
1207-14
1207-15
1207-17*
1207-18
1207-20
1207-21
1207-23
1207-24
1207-26
1207-27
1207-29
1207-30
1st Male
Number
1207-1
1207-1
1207-4
1207-4
1207-7
1207-7
1207-10
1207-10
1207-13
1207-13
1207-16
1207-16
1207-19
1207-19
1207-22
1207-22
1207-25
1207-25
1207-28
1207-28
Fate
B
BB
B
B
B
B
B
BB
B
B
BB
B
B
B
B
B
B
B
B
B
2nd Male
Number
NN
1207-13
NN
NN
NN
NN
NN
1207-7
NN
NN
1207-4
NN
NN
NN
NN
NN
NN
NN
NN
NN
Fate
C
NP
C
C
C
C
C
C
C
C
S
C
C
C
C
C
C
NP
C
C
Days
Pregnant
20
0
20
20
20
20
20
20
20
20
Approx. 18
20
20
20
20
20
20
0
20
20
*Not exposed due to assumption that this animal was not successfully bred.
(Continued)
10
-------
TABLE 5 (Continued)
Key
NN = Not Necessary
BB = Did Not Breed
C = C-Sectioned
S = Sacrificed
NP = Not Pregnant
B = Bred
INDIVIDUAL MATING DATA
Diesel Exhaust Inhalation (Group 2)
Dam
Number
1207-32
1207-33
1207-35
1207-36
1207-38
1207-39
1207-41
1207-42
1207-44
1207-45
1207-47
1207-48
1207-50
1207-51
1207-53
1207-54
1207-56
1207-57
1207-59
1207-60
1st Male
Number
1207-31
1207-31
1207-34
1207-34
1207-37
1207-37
1207-40
1207-40
1207-43
1207-43
1207-46
1207-46
1207-49
1207-49
1207-52
1207-52
1207-55
1207-55
1207-58
1207-58
Fate
B
B
B
B
B
B
B
,B
B
B
B
B
BB
B
B
B
B
B
B
B
2nd Male
Number
NN
NN
NN
NN
NN
NN
NN
NN
NN
NN
NN
NN
1207-31
NN
NN
NN
NN
NN
NN
NN
Fate
C
C
C
C
C
C
C
C
C
C
C
C
NP
C
C
C
C
C
C
C
Days
Pregnant
20
20
20
20
20
20
20
20
20
20
20
20
0
20
20
20
20
20
20
20
11
-------
LAPAROTOMY DATA
Diesel exhaust emissions produced no malformation or other teratogenic effects in the
unborn rat fetuses.
All rat pups were examined at the time of cesarean section and no gross abnormalities
were noted. The bruised areas recorded were due to manipulation of the pups with
forceps (Table 6).
The total number of fetuses, number of live and dead fetuses, number of resorptions,
number of implantation sites, corpora lutea and distribution of fetuses are presented in
Tables 9 and 10. None of these parameters were altered foUowing exposure to diesel
exhaust emissions.
The test material produced no effect on individual pups or total litter weight nor sex
distribution (Tables 7 and 8).
Soft tissue examination of fetuses is presented in Table 11 and no malformations directly
related to diesel exhaust emission exposure were observed. All abnormalities observed
were considered to be slight modifications from normal and were within the limits of
normal variation.
No skeletal malformations related to the test material were noted (Tables 12 and 13).
Skeletal examination of the fetuses revealed some minor spontaneous malformations in
both the control and test groups but these changes were not related to the exposure to
the diesel exhaust emission.
No signs of maternal toxicity were observed during the gross necropsy of the dams but
the lungs of diesel exhaust exposed animals were observed to be a pinkish-brown color as
compared to the pink color in control rats. It was assumed that this change is due to the
presence of test material in the lungs or at least due to a response to the presence of the
diesel exhaust emissions (Table 15).
FETAL VISCERAL DATA
Table 11 presents soft tissue examinations of the fetuses. The only fetuses listed in this
table are the anomalies or abnormalities.
No significant or conclusive data can be made relative to the test material. All fetuses
not listed were normal.
FETAL SKELETAL DATA
Tables 12 and 13 present the mean values of the ossified elements for the term fetuses
at the time of the skeletal examinations.
Table 14 presents the skeletal examination of the fetuses which show anomalies or
abnormalities. All other fetuses were observed to be normal.
The results from these tables have not drawn any conclusive data that can be made
relative to the test material.
12
-------
GROSS NECROPSIES
Table 15 presents the gross necropsy findings from the dams in each group. The only
significant observation made for Group 2, the diesel fuel group, was the distinct pink-
brown appearance of the lungs.
13
-------
TABLE 6
GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY
Dam No.
1207-2
1207-3*
1207-5
1207-6
1207-8
1207-9
1207-11
1207-12
1207-14
1207-15
1207-171
1207-18
Fetus
No. Exposure Level Sex
4 Clean Air M/
Clean Air
12 Clean Air F/
14&15 Clean Air F/F
Clean Air
Clean Air
2 Clean Air M/
12 Clean Air F/
Clean Air
Clean Air
Clean Air
Gross Observation
Left rear leg had 2 mm red area,
also a dark subcutaneous spot was
present on right side midway be-
tween shoulder and ear.
All fetuses appeared normal.
Intradermal blotchy area .5 cm x .5
cm on right side of skull behind ear.
Fetuses were in the same placenta
and they appeared normal.
All fetuses appeared normal.
All fetuses appeared normal.
Subcutaneous bruised area under
chin.
Subcutaneous 2 mm bruised area
along thoracic vertebrae.
All fetuses appeared normal.
All fetuses appeared normal.
All fetuses appeared normal
*Animal was not pregnant
Rat was not exposed due to miscalculation of pregnancy.
(continued)
14
-------
TABLE 6 (Continued)
Dam No.
1207-20
1207-21
1207-23
1207-24
1207-26
1207-27*
1207-29
Fetus
No
3
4
6
6
8
10
10
2
8
10
12
13
15
Exposure Level
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Sex
M/
M/
F/
M/
M/
F/
M/
M/
M/
M/
M/
F/
F/
Gross Observations
Intradermal dark red area on upper
portion of back.
Medial side of left hind leg and
areas between toes were bruised.
Right hind foot bruised on heel area.
Dorsal region of neck has three 1
mm red areas.
A 2 mm red area located on right
side of head between eye and ear.
A 2 mm red area located on right
side of head between eye and ear.
Intradermal area 4 mm x 4 mm
located on right side of head.
All fetuses appeared normal.
Red area 2 mm above ear on right
side of head.
Red area 1 mm located directly
below nose.
Red area 2mm above right eye.
Red area 4 mm midline of snout
also bottom of left front foot.
Red area 2mm above right ear.
Medial side of rear legs are red-
dened.
*Animal was not pregnant.
(continued)
15
-------
TABLE 6 (Continued)
Dam No.
1207-30
1207-30
1207-32
1207-33
1207-35
1207-36
1207-38
1207-39
1207-41
1207-42
1207-44
1207-45
1207-47
Fetus
No
3
7
10
13
12
14
15
17
12
5
6
11
Exposure Level
Clean Air
Clean Air
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Sex
F/
F/
M/
M/
M/
M/
F/
F/
M/
F/
F/
F/
Gross Observations
Intradermal scattered red areas on
left side of head.
Fetus born dead, very small in size,
fluid subcutaneous over entire body
and two 2 mm intradermal red areas
located on back of head.
All fetuses appeared normal.
Bruised area on right side of head.
1 mm red spot midway on tail.
All fetuses appeared normal.
Subcutaneous 1.5 mm red area at
dorsal midline.
All fetuses appeared normal.
Forcep bruises on mouth.
Forcep bruises on mouth.
All fetuses appeared normal.
Intradermal blotchy area 9 mm in
diameter on right side of head.
All fetuses appeared normal.
Bruised area on right side of head.
Bruised areas on right side of head,
thought to be caused from handling.
Bruised area below right ear.
Bruised area on top of nose.
1207-48
Diesel Exhaust
All fetuses appeared normal.
(continued)
16
-------
Dam No.
1207-50*
1207-51
1207-53
1207-54
1207-56
TABLE 6 (Continued)
Fetus
No Exposure Level Sex Gross Observations
3 Diesel Exhaust M/
16 Diesel Exhaust M/
Diesel Exhaust
Diesel Exhaust
7 Diesel Exhaust M/
Fetus appeared to be bloated sub-
cutaneously, also there were subcu-
taneous bruises at neck, head and
lower back region.
Intradermal bruise on top of head.
All fetuses appeared normal.
All fetuses appeared normal.
Intradermal 1 mm dark red area at
1207-57
1207-59
1207-60
8
9
5
16
10
15
1
4
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
M/
F/
M/
M/
F/
F/
M/
F/
neck and mid-back and fissures of
cranium.
Intradermal 1 mm dark red area at
lateral region of left leg.
Intradermal 3 mm x 1 mm dark red
area at cranium.
Bruised area on left side of snout.
Medial side of right hind femur had
a diffused red area, also two red
areas 1 mm each on upper region of
back.
Intradermal red area at end of tail.
Skin on back was cut due to hand-
ling.
4 mm cut on right hip was due to
handling.
Intradermal 2 mm red area on right
side of head.
*Animal was not pregnant.
17
-------
TABLE 7
FETAL DATA FOR DAMS WHICH WERE EXPOSED TO CLEAN AIR (GROUP I CONTROL)
SEX DISTRIBUTION, AVERAGE FETAL WEIGHT (g), GROUP FETAL WEIGHT,
INDIVIDUAL FETUS WEIGHT (g), FETUS POSITION IN UTERINE HORN1
Dam No.
1207-2
1207-32
1207-5
1207-6
1207-8
1 207-9
1207-11
1207-12
1207-14
1207-15
0° 1207-173
1207-18
1207-20
1207-21
1207-23
1207-24
1 207-26
1207-272
1207-29
1207-30
Sex Distribution
Males Females
9
2
9
4
5
7
7
9
6
7
9
7
9
7
6
10
4
6
4
6
11
5
6
5
5
5
7
5
7
4
6
7
6
6
Average
Total Number Fetal
of Fetuses Wt. (g)
15
6
15
15
10
13
12
14
11
14
H
14
13
13
13
16
10
3.7
3.8
3.7
3.8
3.8
4.1
3.7
3.6
3.9
4.0
3.7
3.5
4.1
3.9
3.4
3.9
3.4
Group
Fetal
Wt.(g) 1
56.2
22.5
54.8
56.7
38.2
53.7
44.0
50.4
42.8
55.3
51.1
49.2
52.8
50.5
44.3
61.8
34.1
3.4
3.8
3.6
3.7
*
4.1
3.6
3.6
3.8
3.9
3.5
3.6
*
3.9
3.3
3.7
3.4
2
4.0
3.8
3.6
3.6
*
4.3
3.6
3.4
4.1
3.7
3.4
3.7
4.3
4.3
3.4
4.1
3.7
3
4.1
3.6
3.7
*
*
4.5
3.8
3.7
4.1
3.8
3.4
4.0
4.1
*
3.6
3.5
3.6
4
3.9
*
3.9
4.2
2.7/
A
3.8
3.5
3.8
3.9
3.7
3.5
4.3
3.5
3.2
3.9
*
5
3.7
4.0
3.7/
3.7
3.9
3.8
3.7
3.57
3.6
4.1
3.8
2.9/
*
4.0
3.37
3.8
*/
6
4.0
3.5
3.3
4.1
3.7
*
3.4
3,8
3.8
4.1
3.8
3.4
3.8/
3.8
3.6
4.1
3.4
7
4.0
3.S7
3.1
3.9/
4.2
4.4
3.5
*
3.9/
4.2/
3.9
3.2
4.1
4.0
3.1
3.3
1.7
8
4.07
3.8
4.0
3.6
4.1
3.97
3.4
3.6
4.3
3.67
3.7
3.9
3.8/
3.3
4.4/
4.2
9
3.7
4.1
3.8
4.2
4.7/
3.5
3.6
3.8
*
3.6
3.6
3.9
3.9
3.3
3.9
3.8
10
3.6
3.8
3.7
3.8
4.2
3.6
3.8
4.0
3.8
3.5
3.4
4.1
3.7
3.3
4.0
3.2
11
3.4
*
3.8
4.3
4.4
*
3.7
4.3
4.4
3.6
3.8
3.9
3.8
3.5
3.4
4.1
12
3.5
3.3
3.7
3.9
3.7
3.7
3.8
4.0
3.8
3.5
4.2
3.9
3.9
4.1
*
13
3.5
4.2
3.9
3.9
3.8
4.0
*
3.8
3.7
3.5
3.9
3.9
3.5
3.7
3.0
14 15 16 17
3.6 3.8
3.6 3.5 3.6
3.4 3.5 3.7
4.1 3.6
3.6 * 3.8 3.2
3.7 3.6
3.8
3.4
4.2 4.1
4.0
4.1 3.8 4.0
*
1 Positions are counted from the left Fallopian Tube to the right Fallopian Tube.
2 Not pregnant
3 Female was sacrificed due to miscalculation of pregnancy and non-exposure, only number of fetuses and corpora lutea were counted. Weight, sex, and position
of fetus were not recorded.
* Early resorption
/ Denotes position of cervix
-------
TABLE 8
FETAL DATA FOR DAMS WHICH WERE EXPOSED TO DIESEL EXHAUST INHALATION (GROUP II - TEST)
SEX DISTRIBUTION, AVERAGE FETAL WEIGHT (g), GROUP FETAL WEIGHT,
INDIVIDUAL FETUS WEIGHT (g), FETUS POSITION IN UTERINE HORN1
Dam No.
1207-32
1207-33
1207-35
1207-36
1207-38
1207-39
1207-41
1207-42
1207-44
1207-45
1207-47
1207-48
1207-502
1207-51
1 207-53
1207-54
1207-56
1207-57
1207-59
1 207-60
Sex Distr
Males
5
6
4
8
8
7
5
9
6
9
4
6
11
7
8
8
5
7
7
ibution
Females
5
7
8
8
9
8
8
6
7
5
8
6
6
6
9
8
11
8
8
Total Number
of Fetuses
10
13
12
16
17
15
13
15
13
14
12
12
17
13
17
16
16
15
15
Fetal
Wt. (g)
3.5
3.5
3.7
3.4
4.2
3.6
3.6
3.6
3.9
3.9
3.5
3.5
3.4
3.9
3.7
3.6
3.5
3.3
3.8
Fetal
Wt.(g)
34.5
45.5
43.9
55.1
71.3
54.5
47.1
53.5
50.1
54.1
41.6
41.8
57.9
50.4
63.7
59.3
56.6
49.0
57.2
1
3.0
3.5
3.9
3.4
4.4
3.3
3.9
3.2
4.2
4.0
3.6
3.4
4.0
3.7
3.6
3.2
3.6
3.1
3.9
2
*
3.4
3.6
3.3
4.2
*
3.8
*
4.1
4.1
3.4
3.6
3.3
3.8
3.9
3.7
3.6
3.3
3.9
3
3.8
3.4
4.2
3.1
3.9
3.7
3.7
3.5
3,9
4.2
3.3
3.6
4.0
4.0
3.8
3.7
3.4
3,3
4.0
4
*
3.5
3.5
3.8
4.3
3.7
3.3
3.6
4.0
3.S7
2.6
3.5
3.3
3.7
3.7
3.9
3.6
3.7
3.7
5
*
3.57
3.8/
3.4
3.8
3.7
3.5
*
3.5
3.9
3.2
3.4
3.2
4.2
3.5
3.9
3.7
3.2
3.8
6
3.7
3.6
A
3.3
4.3
3.77
3.67
4.07
4.07
3.5
3.5
3.4
3.2
4.07
3.4
3.5
3.S7
3.4
4.2/
7
3.2
3.9
3.3
3.57
3.57
3.4
3.3
3.4
3.7
3.3
3.7/
4.0
3.0
3.9
3.8
3.6
3.9
3.0
3.5
8
4.07
3.6
3.6
3.7
4.4
3.2
3.7
3.3
3.7
4.0
3.7
*/
4.0
3.7
3.77
3.77
3.5
3.1
3.5
9
3.6
3.2
3.7
3.6
4.0
3.6
3.8
3.4
3.8
4.1
3.6
A
3.3/
3.6
3.9
3.6
3.4
2.77
4.0
10
A
3.6
4.0
3.4
4.0
3.8
3.7
3.7
3.8
3.9
3.5
3.4
3.6
3.9
3.9
4.1
3.4
2.9
3.8
11
3.3
3.3
3.5
3.5
4.5
3.7
3.3
3.7
3.8
4.1
3.7
3.3
3.0
3.9
3.5
*
3.5
3.7
3.9
12
3.3
3.3
3.3
3.9
4.3
3.7
3.8
3.6
4.0
3.7
3.8
3,6
*
4.1
3.6
3.9
3.3
3.5
3.9
13
3.5
3.7
3.5
2.6
4.4
3.8
3.7
3.8
3.6
3.9
3.3
3.1
3.9
4.1
3.4
3.4
3.7
3.7
14
3.1
3.6
4.5
3.8
3.9
3.6
3.3
3.6
3.7
4.0
3.6
3.4
3.8
15
3.7
4.5
3.7
3.6
3.8
3.9
3.9
3.8
3.0
3.6
16 17 18
3.3
3.9 4.4 *
3.7
3.8 3.0
3.7 2.5 3.3
3.8 3.9
3.6 3.6
3.1
1 Positions are counted from the left Fallopian Tube to the right Fallopian Tube
2 Not pregnant
* Early resorption
/ Denotes position of cervix
-------
TABLE 9
FETAL DATA FOR DAMS WHICH WERE EXPOSED TO CLEAN AIR (GROUP I - CONTROL)
NUMBER OF FETUSES VIABLE AND DEAD, EARLY RESORPTIONS,
IMPLANTATION SITES, AND CORPORA LUTEA
Viable Fetuses
Dam No.
1207-2
1207-31
1207-5
1207-6
1207-8
1207-9
1207-11
1207-12
1207-14
1207-15
1207-172
1207-18
1207-20
1207-21
1207-23
1207-24
1207-26
1 207-27 '
1207-29
1207-30
Left
Horn
8
6
5
6
I
7
8
5
7
1
8
5
4
7
5
8
3
Right
Horn
7
0
10
9
9
6
4
9
4
7
6
9
9
6
8
8
6
Total
15
6
15
15
10
13
12
14
11
14
14
14
13
13
13
16
9
Dead Fetuses
Left
Horn
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Right
Horn
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1
Total
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1
Early Resorption
Left
Horn
0
1
0
1
3
2
0
0
0
0
0
0
2
1
0
0
2
Right
Horn
0
0
1
0
0
0
1
3
0
1
0
0
0
0
0
0
2
Total
0
1
1
1
3
2
1
3
0
1
0
0
2
1
0
0
4
Implantation Sites
Left
Horn
8
7
5
7
4
9
8
5
7
7
8
5
6
8
5
8
5
Right
Horn
7
0
11
9
9
6
5
12
4
8
6
9
9
6
8
8
9
Total
15
7
16
16
13
15
13
17
11
15
14
14
15
14
13
16
14
Corpora Lutea
Left
Horn
8
7
5
8
7
9
8
9
7
7
10
7
7
8
5
8
5
Right
Horn
7
6
11
9
9
6
9
14
4
8
7
12
9
6
8
9
9
Total
15
13
16
17
16
15
17
23
11
15
17
19
16
14
13
17
14
1 Not pregnant
2 Female was sacrificed due to miscalculation of pregnancy and non-exposure.
-------
TABLE 10
FETAL DATA FOR DAMS WHICH WERE EXPOSED TO DIESEL EXHAUST INHALATION (GROUP II - TEST)
NUMBER OF FETUSES VIABLE AND DEAD, EARLY RESORPTIONS,
IMPLANTATION SITES, AND CORPORA LUTEA
Viable Fetuses Dead Fetuses Early Resorption Implantation Sites Corpora Lutea
Dam No.
1207-32
1207-33
1207-35
1 207-36
1207-38
1207-39
1207-41
1 207-42
1 207-44
1 207-45
1207-47
1 207-48
1 207-50 ]
1207-51
1207-53
1207-54
1207-56
1207-57
1207-59
1 207-60
Left
Horn
5
5
5
7
7
5
6
4
6
4
7
7
9
6
8
8
6
9
6
Right
Horn
5
8
7
9
10
10
7
11
7
10
5
5
8
7
9
8
10
6
9
Total
10
13
12
16
17
15
13
15
13
14
12
12
17
13
17
16
16
15
15
Left
Horn
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Right
Horn
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Total
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Left
Horn
3
0
0
0
0
1
0
2
0
0
0
1
0
0
0
0
0
0
0
Right
Horn
1
0
1
0
1
0
0
0
0
0
0
1
1
0
0
1
0
0
0
Total
4
0
1
0
1
1
0
2
0
0
0
2
1
0
0
1
0
0
0
Left
Horn
8
5
5
7
7
6
6
6
6
4
7
8
9
6
8
8
6
9
6
Right
Horn
6
8
8
9
11
10
7
11
7
10
5
6
9
7
9
9
10
6
9
Total
14
13
13
16
18
16
13
17
13
14
12
14
18
13
17
17
16
15
15
Left
Horn
8
7
5
8
7
9
6
7
6
4
12
9
11
6
8
10
7
9
7
Right
Horn
7
10
8
10
11
10
7
11
8
12
7
8
11
7
9
8
10
6
9
Total
15
17
13
18
18
19
13
18
14
16
19
17
22
13
17
18
17
15
16
1 Not pregnant
-------
TABLE 11
SOFT TISSUE EXAMINATION OF THE FETUSES
Dam No.
1207-9
1207-15
1207-20
1207-26
1207-35
1207-44
1207-45
1207-48
1207-60
Fetus
No.
9
3
6
9
9
12
3
7
3
6
9
3
3
11
6
9
Exposure Level
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Clean Air
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Sex
M/
M/
M/
M/
M/
M/
M/
F/
F/
F/
M/
M/
M/
F/
M/
M/
Abnormality
Kidneys show slight nephredema.
Kidneys show slight nephredema.
Left kidney showed slight nephr-
edema and left ureter was dilated.
Right testicle higher than left.
Right testicle retained.
Right testicle sitting on rectum and
leaning to the left.
Left testicle higher than right.
Left uterine horn shorter in length.
Right kidney had nephredema.
Right kidney had nephredema.
Ureters were dilated.
Kidneys showed nephredema.
Kidneys showed nephredema and
ureters were anteriorly dilated.
Right kidney showed nephredema
and right ureter anteriorly dilated.
Right kidney showed nephredema.
Left kidney showed nephredema and
left ureter anteriorly dilated. Left
testicle was cryptorchid.
22
-------
ro
CO
TABLE 12
SKELETAL EXAMINATION OF TERM FETUSES, MEAN NUMBER OF
OSSIFIED ELEMENTS - CLEAN AIR INHALATION (GROUP I)
Dam No: 2 5 6 8 9 11 12 14
r>
0
D
3°
c
-------
TABLE 12 (Continued)
Dam No: 15 18 20 21 23 24 26 29* 30
Cervicle
Thoracic
Lumbar
Sacral
Tail
Sternum
Ribs
Foredigits
Metacarpals
Proximal
1.)
Middle 2.) -
3.)
Distal
Hindigits
Metacarpals
Proximal
1.)
Middle 2.) -
3.)
Distal
6
13
6
6
2.1
5.9
13
L
3.3
1
3
1
L
4
1
3
1
R
3.3
1
3
1
R
4
1
3
1
1
5.
1
L
3
1
3
1
L
4
1
3
1
6
3
6
6
2
6
3
R
3
1
3
1
R
4
1
3
1
1
2.
1
L
4
1
2
1
L
4
1
3
1
6
3
6
6
5
6
3
R
4
1
2
1
R
4
1
3
1
2
5
L
3.8
1
3
1
L
4
1
3
1
6
13
6
6
.4
.5
13
R
3.8
1
3
1
R
4
1
3
1
5
L
3.8
1
2.8
1
L
4
1
3
1
6
13
6
6
2
.9
13
R
3.8
1
2.8
1
R
4
1
3
1
L
4
1
3
1
L
4
1
3
1
6
13
6
6
3
6
13
R
4
1
3
1
R
4
1
3
1
6
13
6
6
2.8
5.9
13
L R
4 4
1 1
2 2
1 1
L R
4 4
1 1
3 3
1 1
6
13
6
6
2.8
5.9
13
L
3.5 3
1
3
1
L
41
I1
31
I1
R
.5
1
3
1
R
44
I4
34
I4
6
12.7
6
5.6
1.7
4.6
12.7
L R
2.7 2.9
.9 .9
2.6 2.6
.9 .9
L R
3.4 3.4
.9 .9
2.6 2.6
.9 .9
1 One Fetus1 left hind foot missing due to handling, therefore, these mean values are exclusive of this foot.
4 One Fetus1 right hind foot was missing due to over masceration, therefore, mean value is exclusive of this foot.
* One whole fetus was not included in mean values due to over masceration.
-------
IV>
cn
TABLE 13
SKELETAL EXAMINATION OF TERM FETUSES, MEAN NUMBER OF
OSSIFIED ELEMENTS - DIESEL EXHAUST INHALATION (GROUP II)
Dam No.: 32 33 35 36 38 39
n
o
3
"•?•
3
C
CD
Q_
Cervicle
Thoracic
Lumbar
Sacral
Tail
Sternum
Ribs
Foredigits
Metacarpals
Proximal
i \
I. j
Middle 2. ) -
3. )
Distal
Hindigits
Metacarpals
Proximal
1. )
Middle 2. ) -
3. )
Dista!
6
13
6
6
1.9
5.6
13
L
3.3
1
2
1
L
4
1
3
1
R
3.3
1
2
1
R
4
1
3
1
6
13
6
6
2
5.7
13
L
3.3
1
2
1
L
4
1
3
1
R
3.3
1
2
1
R
4
1
3
1
6 6
13 13
6 6
6 6
3.4 2.5
6 5.1
13 13
L R L
4 4 3.5
1 1 1
22 3
1 1 1
L R L
444
1 1 1
333
1 1 1
R
3.5
1
3
1
R
4
1
3
1
6
13
6
6
3.2
5.5
13
L
3.8
1
2
1
L
4
1
3
1
R
3.8
1
2
1
R
4
1
3
1
6
13
6
6
2.5
5.7
13
L
3.7
1
2.8
1
L
4
1
2.9
1
R
3.8
1
2.8
1
R
4
1
2.9
1
-------
TABLE 13 (Continued)
SKELETAL EXAMINATION OF TERM FETUSES, MEAN NUMBER OF
OSSIFIED ELEMENTS - DIESEL EXHAUST INHALATION (GROUP II)
Dam No.:
41
42
44
45
47
48
Cervicle
Thoracic
Lumbar
Sacral
Tail
Sternum
Ribs
"n Foredigits
3 Metacarpals
rv> D* Proximal
en c i \
a> 1 . )
~ Middle 2. ) -
3. )
Distal
Hindigits
Metacarpals
Proximal
1. )
Middle 2. ) -
3. )
Distal
6
13
6
6
2.4
5.1
13
L
3.6
1
2
1
L
4
^9
2.7
1
R
3.7
1
2
1
R
4
.9
2.7
1
6
13
6
6
2
4.7
13
L
3
1
2
1
L
4
1
3
3
R
31
1 1
21
I1
R
4
1
3
3
6
13
6
6
2.6
5.3
13
L
3.8
1
3
1
L
4
1
3
1
R
3.8
1
3
1
R
4
1
3
1
6
13
6
6
2.2
5.4
12.9
L
3.6
1
3
1
L
4
1
3
1
R
3.6
1
3
1
R
42
I2
32
I2
6
13
6
6
1.4
5.3
13
L
3
1
3
1
L
3.9
1
3
1
R
3
1
3
1
R
3.9
1
3
1
6
13
6
6
2.1
5.5
13
L
3.5
1
3
1
L
4
1
3
1
R
3.6
1
3
1
R
4
1
3
1
1 One Fetus1 right hand was missing due to over masceration, therefore, these mean vglues are exclusive of this hand.
2 One Fetus1 hind foot was missing due to handling, therefore, these mean values are exclusive of this foot.
-------
TABLE 13 (Continued)
SKELETAL EXAMINATION OF TERM FETUSES, MEAN NUMBER OF
OSSIFIED ELEMENTS - DIESEL EXHAUST INHALATION (GROUP II)
Dam No.:
Cervicle
Thoracic
Lumbar
Sacral
Tail
Sternum
Ribs
Foredigits
Metacarpals
Proximal
1. )
Middle 2. ) -
3. )
Distal
Hindigits
Metacarpals
Proximal
1. )
Middle 2. ) -
3.)
Distal
2
5
L
3.2
1
3
1
L
4
1
3
1
51
6
13
6
6
.1
.3
13
R
3.
1
1
3
1
R
4
1
3
1
53
6
13
6
6
3
5.9
13
L
3.8
1
2.2
1
L
4
1
3
1
R
3.8
1
2.3
1
R
4
1
3
1
54
63
13
6
6
2.3
5.3
13
L
3.2
1
3
1
L
3.8
1
3
1
i
R
3.3
1
3
1
R
3.9
1
3
1
56
6
13
6
5.9
2.6
6
13
L
3.7
1
2.6
1
L
4
1
3
1
R
3.7
1
2.6
1
R
4
1
3
1
57
6
13
6
6
1.6
5.1
13
L R
3 3
1 1
3 3
1 1
L R
4 4
1 1
3 3
1 1
59
6
13
6
6
1.6
5
13
L
3
1
2.3 2.
1
L42
I2
2
2.4 2.
I2
R
3
1
,3
1
R
4
1
7
1
60
6
13
6
6
1.7
5.2
13
L
3.2
1
3
1
L
4
1
3
1
R
3.2
1
3
1
R
4
1
3
1
2 One Fetus1 hind foot was missing due to handling, therefore, these mean values are exclusive of this foot.
3 One Fetus1 cervicle area was beyond recognition, therefore, this mean value was exclusive of this particular
Fetus1 cervical bones.
-------
TABLE 14
SKELETAL EXAMINATION OF THE FETUSES
Dam No.
1207-6
1207-8
1207-14
Fetus
No.
1207-18
1207-29
1207-30
1207-36
15
11
12
15
13
10
Exposure Level Sex
Clean Air F/
Clean Air F/
Clean Air M/
Clean Air M/
Clean Air F/
Abnormality
Clean Air
Clean Air
Clean Air
Clean Air
M/
M/
F/
M/
Diesel Exhaust F/
(continued)
Sternum was not aligned. Stern-
ebrae 5 and 6 appeared to be shifted
to the left.
All five upper sternebrae appear to
be split and not aligned. 6th stern-
ebrae appears to be normal.
Left lumbar 1 appeared to have an
extension.
Second sternebrae was not com-
pletely calcified.
Parietal bone in skull was difficient
and parietal frontal bones were
deficient.
Sternebrae 3, 4, 5, and 6 were not
aligned.
Sternebrae 2 and 5 were not com-
pletely calcified.
Fetus was abnormally small and
immature, bone structure was not
complete.
Intraparietal bone of skull was not
completely calcified and the supra-
occipal was not symmetrical.
Sternebrae 2 was not completely
calcified.
Supraoccipital of skull was not
completely calcified and was non-
symmetrical.
28
-------
TABLE 14 (Continued)
Dam No.
1207-36
1207-38
1207-42
1207-51
1207-54
1207-54
Fetus
No
13
10
9
Exposure Level Sex
Abnormality
10
17
11
13
16
10
16
1207-56
Diesel Exhaust M/
Diesel Exhaust F/
Diesel Exhaust M/
Diesel Exhaust M/
Diesel Exhaust M/
Diesel Exhaust F/
Diesel Exhaust M/
Diesel Exhaust F/
Diesel Exhaust M/
Diesel Exhaust F/
Diesel Exhaust M/
Sternebrae 1, 3, 5 and 6 were not
present.
Parietal bones appeared to be
slightly separated.
Parietal bones appeared to be
slightly separated and all cranial
bones appeared irregular and lack
symmetry.
Parietal bones appeared to be
slightly separated and all cranial
bones appeared irregular and lacked
symmetry.
Second sternebrae was not com-
pletely calcified.
Right and left femur at distal end
was darker in color.
Left humerus at distal end was
darker in color.
Left femur at distal end was darker
in color.
Sternebrae 3 was split.
Sternebrae was difficult to deter-
mine, some appeared to be split.
Sternum appeared to have 7 stern-
ebrae, however, the first one may
have been split.
29
-------
TABLE 15
GROSS NECROPSY RESULTS FOR 20-DAY LAPAROTOMY FOR FEMALE RATS
Dam No.
1207-15
1207-23
Group
No.
1207-32
1207-33
1207-35
1207-36
1207-38
1207-39
1207-41
1207-42
1207-44
2
2
2
2
2
2
2
2
Exposure Level
Clean Air
Clean Air
Clean Air
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Gross Necropsy Findings
Kidneys appeared mottled. No other remark-
able gross pathology was noted.
Lungs were reddened and diffused. Slightly
brown. No other remarkable gross pathology
was noted.
No remarkable gross pathology was noted in any
female rat from this group.
Lungs were found to be pinkish-brown in color.
No other remarkable gross pathology was noted.
No remarkable gross pathology was noted.
Lungs were found to be pinkish-brown in color.
No other remarkable gross pathology was noted.
Lungs were found to be pinkish-brown in color.
No other remarkable gross pathology was noted.
Lungs were found to be pinkish-brown in color.
No other remarkable gross pathology was noted.
Lungs were pinkish-brown in color. No other
remarkable gross pathology was noted.
Lungs were pinkish-brown in color. No other
remarkable gross pathology was noted.
Lungs were mottled dark red on all lobes. No
other remarkable gross pathology was noted.
No remarkable gross pathology was noted.
(continued)
30
-------
Group
Dam No. No. Exposure Level
TABLE 15 (Continued)
Gross Necropsy Findings
1207-45
1207-47
1207-48
1207-50
1207-51
1207-53
1207-54
1207-56
1207-57
1207-59
1207-60
2
2
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Diesel Exhaust
Lungs appeared to be brown and red in color.
No other remarkable gross pathology was noted.
No remarkable gross pathology was noted.
Lungs were mottled dark red except for edges
of all lobes which were pink. No other remark-
able gross pathology was noted.
Lungs were pinkish-brown in color. No other
remarkable gross pathology was noted.
Lungs appeared brownish-red in color. No other
remarkable gross pathology was noted.
Lungs were mottled red in color. No other
remarkable gross pathology was noted.
Lungs appeared to be pinkish-brown in color.
No other remarkable gross pathology was noted.
Lungs appeared brown; right and left diaphrag-
matic lobes of the lungs were red at edges. No
other remarkable gross pathology was noted.
Lungs appeared pink with large areas of red on
every lobe. No other remarkable gross path-
ology was noted.
Lungs appeared pinkish-brown in color. No
other remarkable gross pathology was noted.
Lungs appeared pinkish-brown in color. No
other remarkable gross pathology was noted.
31
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing}
1. REPORT NO.
EPA-600/1-80-010
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
TERATOLOGIC EFFECTS OF LONG-TERM EXPOSURE
TO DIESEL EXHAUST EMISSIONS (RATS)
5. REPORT DATE
January 1980 issuing date
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Kathleen M. Werchowski, A.S., Vincent W. Chaffee, D.V.M.
G. Bruce Briggs, D.V.M.
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
WIL Research Laboratories, Inc.
3154 Exon Avenue
Cincinnati, Ohio 45241
10. PROGRAM ELEMENT NO.
11. CONTRACT/GRANT NO.
68-03-2652
12. SPONSORING AGENCY NAME AND ADDRESS
Health Effects Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, Ohio 45268
13. TYPE OF REPORT AND PERIOD COVERED
Teratologic 09-15-78 to 02-07-79
14. SPONSORING AGENCY CODE
EPA/600/10
15. SUPPLEMENTARY NOTES
16. ABSTRACT
This research project was initiated with the objective of evaluating the
potential for diesel exhaust emissions to produce malformations in rat
fetuses. The dams were exposed by the inhalation route to a 10% concen-
tration of diesel exhaust emissions in inhalation chambers on days 6 through
15 of gestation. Methods used conform to the guideline developed by the
Food and Drug Administration for evaluating teratogenic effects in rats.
The results of the exposure of pregnant rats indicate that diesel exhaust
emissions have no effect upon the normal development of rat fetuses.
Diesel exhaust emissions did not effect any of the parameters evaluated
to assess maternal toxicity nor total number of fetuses born.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS C. COSATI Field/Group
18. DISTRIBUTION STATEMEN1
19. SECURITY CLASS (This Report)
Release to Public
21. NO. OF PAGES
40
20 SECURITY CLASS (This page)
Unclassified
22. PRICE
EPA Form 2220-1 (Rev. 4-77) PREVIOUS EDITION is oesoi_ETE32
t U S GOVERNMENT PRINTING OFFICE 1980-657-146/5571
------- |