United States
Environmental Protection
Agency
Office of Health and
Environmental Assessment
Washington DC 20460
Research and Development
EPA/600/S6-87/006  Aug. 1988
Project  Summary
Carcinogenicity Assessment of
Aldrin  and Dieldrin

Finis L. Cavender, Brion T. Cook, and Norbert P. Page
  Evidence pertaining to the carcino-
genicity of aldrin/dieldrin is reviewed
and evaluated. The full report covers
studies completed before 1985. Case
reports and epidemiologic  studies of
pesticide  applicators  and pesticide
manufacturing workers are reviewed,
but because of methodologic limita-
tions, these studies established neither
a positive nor a negative association
between cancer and  aldrin/dieldrin
exposure.  A number of independent
studies of laboratory animals, however,
demonstrated that aldrin/dieldrin
cause liver cancer in mice and rats.
Based on  the accumulated evidence,
aldrin/dieldrin are classified as proba-
ble human carcinogens. Group B2.
using EPA's Guidelines for Carcinogen
Risk Assessment. The carcinogenic
potency of aldrin/dieldrin is estimated
by fitting mathematical models to the
laboratory  animal data. These esti-
mates indicate that aldrin/dieldrin are
rather potent carcinogens,  ranking in
the first (most potent) quartile of
potential  carcinogens  evaluated by
EPA's Carcinogen Assessment Group.
Separate mutagenicity assessments of
aldrin/dieldrin are attached as appen-
dices to the full report. The full report
also includes an extensive list of
references pertinent to the carcinogen-
icity of aldrin/dieldrin.
  This Project Summary was devel-
oped by EPA's Office of Health and
Environmental Assessment, Washing-
ton, DC, to announce key findings of
the research project that is fully doc-
umented in a separate report of the
same title (see Project Report ordering
information at back).

Introduction
  Aldrin and dieldrin were first synthe-
sized  in the laboratory  in  1948  with
commercial  production in  the  United
States first reported in 1950. Aldrin and
dieldrin are organochlorine pesticides
extensively used in the 1960s and early
1970s for a large variety of pesticidal
uses.  Most  uses were banned  in the
United States in 1974 under the Federal
Insecticide, Fungicide,  and  Rodenticide
Act. Uses were also restricted in many
other countries at about the same time.
The approved uses in the United States
now are mainly for termite control by
direct soil injection and for nonfood seed
and plant treatment. Production of aldrin
and dieldrin in the United States was also
discontinued along with the restrictions
of 1974, so that virtually all aldrin and
dieldrin now used in the United States
is  imported. Exposure of humans has
been primarily from dermal  and inhala-
tion exposure related to the application
of pesticides; however, due to the
persistence and  bioaccumulation of
dieldrin, considerable exposure  has
occurred through  ingestion  of contam-
inated water and food products. In
contrast, little human exposure from
environmental sources of aldrin occurs,
as it is readily converted to dieldrin by
direct expoxidation in the environment.
During the early 1970s,  nearly all
humans sampled had measurable tissue
levels of dieldrin and, due to its stability
and persistence, dieldrin is still detected
in humans.  However, the levels are
considerably low. Aldrin and dieldrin are
usually considered together, as they are
structurally  related cyclodiene insecti-
cides, have  similar uses, and because
aldrin is readily epoxidized to dieldrin in
the body or the environment.

  The full report reviews the currently
available literature on the  toxicity and
carcinogenicity of both aldrin and dieldrin
and conducts an assessment as to the

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potential carcinogenicity risk to humans.
The  Carcinogen Assessment  Group
(CAG) conducts a "weight-of-evidence"
evaluation that weighs data concerning
innate  biological activity  including
metabolism, toxic effects as related to
duration and levels of exposures, results
of mutagenicity  and  other short-term
tests, long-term animal bioassays, and,
where  available, data derived from
epidemiologic studies. Since mutagenic-
ity is considered as one major mecha-
nism for cancer induction, those studies
are considered supportive of the in vivo
animal or human data.
  Following a review of the data, those
data most appropriate for mathematical
extrapolation are selected for risk assess-
ment. Several mathematical .models, as
described  in EPA's Guidelines for Car-
cinogen Risk Assessment are used. On
the basis  of the "weight-of-evidence"
and risk extrapolation, the substance is
classified as a potential human  carcin-
ogen according to the EPA Guidelines.


Discussion

Human Studies
  Six case reports on aldrin  toxicity
describe neurotoxicity, including convul-
sions and  abnormal EEGs, as the main
toxic effects. Five case reports were
found for dieldrin. Like aldrin, effects of
the central nervous  system, including
convulsions, were the main signs follow-
ing acute or prolonged exposure. In one
of the reports, however, immunohemo-
lytic anemia was the primary toxic effect.
  Only  two epidemiologic studies of
humans exposed to  aldrin or  dieldrin
were reported.  One was  a  long-term
study  of  233  industrial workers at
pesticide manufacturing and formulating
plants in the Netherlands. However, the
study focused mainly on clinical param-
eters and  not cancer,  and is considered
inadequate to determine the carcinogen-
icity of aldrin and dieldrin.  The other
study included a retrospective mortality
analysis of 1,155 employess at  a plant
that had produced aldrin,  dieldrin,  and
endrin for 30 years prior to the analysis.
There  was a statistically significant (p
<0.01) increase in nonmalignant respi-
ratory  disease  (SMR=212, p<0.01).
While  an  SMR of 82 was found for all
malignant neoplasms, an SMR of  235
was found for cancer of the esophagus,
SMR=242 for  cancer of the  rectum,
SMR=225  for cancer of the liver, and
SMR=147 for cancer of the lymphatic and
hematopoietic system. However, none of
these was significnat. This study also has
serious limitations. No information was
presented on actual exposure levels, and
exposures  to other chemicals  were
known to occur. No attempts were made
to adjust for  effects of other  chemical
exposures,  smoking,  or  alcohol  con-
sumption. Since the number of workers
was small and vital status was  unknown
for 10%, the  power to detect  an  effect
was quite limited.  Due to  these  meth-
odological limitations  and the limited
data, it is  difficult to draw  either  a
negative or positive association between
aldrin/dieldrin  and carcinogenicity.
Hence,  these studies  are considered
inadequate epidemiologic evidence.

Animal Studies

Aldrin
  Three adequately conducted long-term
carcinogenicity bioassays of aldrin have
been conducted with mice. The strains
used were C3HeB/Fe, C3H, and B6C3Fi.
In the study with B6C3Fi mice, aldrin was
fed at four doses of 3 to 8 ppm for 80
weeks with 10 to 13 weeks of additional
observation. A statistically significant (p
<0.001) increase  in  hepatocellular
carcinomas was observed in both the  4
and 8 ppm  male groups but not  in the
females. In  the other two  studies,
reported  in 1962 and  1965,  C3H and
CaHeB/Fe mice were fed aldrin at 10 ppm
for 24  months. In both, statistically
significant (p <0.001) increased inciden-
ces of  hepatomas were diagnosed
although not broken down as to inciden-
ces in males or females. Reevaluation by
other pathologists diagnosed the lesions
as hepatocellular carcinomas  with sta-
tistically significant (p <0.05) increases
in both male and female groups.
  While it may be true that the CaH and
CaH/HeB/Fe  strains have  a large per-
centage of their gene pool in  common,
we expect that B6C3Fi mice have approx-
imately half of the gene pool of the C3H
strain. Furthermore, the gene pool in
these mice changes from lab to lab and,
after  many generations,  in the  same
laboratory. The fact  that  dieldrin,  a
metabolite of aldrin, is carcinogenic in
the C57BL/6J, CBA/J, Swiss-Webster,
and CF-1 strains (highly different genet-
ically) as well as  the C3  and B6C3F,
strains argues against the C3H strain's
unique  genetic  susceptibility to this
effect.
  Seven  carcinogenicity  bioassays of
aldrin have been conducted with rats,
only one of which is considered adequate
in design and  conduct. Five studies used
Osborne-Mendel rats,  with  one  each
using Carworth and Holtzman strains.
Doses ranged from 0.5 to 150 ppm with
dietary exposure generally for at least 24
months. At  50 ppm and  below,  good
survival resulted. There was no evidence
of carcinogenicity in any of the studies.
However, a  constant finding  was liver
lesions comonly referred to as "chlori-
nated insecticide type lesions."
  In the only adequate study,  the
National Cancer Institute (NCI) bioassay
in Osborne-Mendel rats, there were
increased incidences of thyroid follicular
cell adenomas  and carcinomas of the
thyroid in both males and females. These
incidences were significant in the low-
dose group but not the high-dose group
when compared to  pooled controls but
not with matched controls. It should be
noted that there were only 10 animals
in the matched controls, which is not
usually enough to  detect all but the
largest responses.  Although  NCI  con-
cluded that the tumors were  not asso-
ciated with  treatment, our examination
using pooled controls indicates that there
was some  positive response.  Later
evaluations by other investigators indi-
cated that the thyroid follicular (in both
sexes) and  adrenal cortex  tumor in
female rats was suggestive evidence of
carcinogenicity for aldrin.
  Another study with rats was consid-
ered inadequate because as few as 62%
of the treated animals in one dose group
(female high-dose group) and 75% of the
male controls were microscopically
examined. Reevaluation of the histopath-
ology indicated that the authors may
have under estimated and underreported
the incidence of malignant  tumors by
approximately threefold. In   all, there
were seven rat studies. Six of them were
considered inadequate.  A well-designed
study on rats would be useful to answer
the specific  question whether the car-
cinogenic potential of aldrin is limited to
mice or not.


Dieldrin
  Twelve carcinogenicity  bioassays of
dieldrin have been conducted with mice;
all  but one  were  judged adequate in
design. Seven strains of mice were used:
CaHeB/Fe, C3H, CBA/J, Swiss-Webster,
CF,, B6C3F1( and C57BL/6J.  In all
studies, either benign or malignant liver
tumors were observed.  The  authors of
six of these  studies indicated that the
tumors were malignant (hepatocellular
carcinomas), whereas in five others the
tumors were diagnosed as hepatomas.

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In three of these five, a  pathology
reevaluation was  performed by other
pathologists who classified the lesions
as being malignant. In three studies, all
with CFi mice,  many of the  hepatocel-
lular carcinomas had metastasized to the
lung. In additon, in one study, a signif-
icant (p <0.05)  reduction  in  latency
period was observed. In one of the
studies with CF1 mice, a  slight but
significant  (p <0.05)  increase  in the
incidence of pulmonary adenomas and
carcinomas, lymphoid tumors, and other
tumors  was seen  in female mice at 1
ppm. The doses used in  the  bioassays
ranged from 0.1  to 20 ppm with dietary
administration for 18 to 24 months.
  Seven long-term carcinogenicity stud-
ies were conducted with rats, three of
which were considered adequate carcin-
ogenicity assays.  The  others suffered
mainly from too few animals, too high
mortality, too short a duration, and/or
inadequate pathology  examination or
reporting. Four  strains  were used: Car-
worth, Osborne-Mendel, Holtzman, and
Fischer 344. Doses ranged from 0.1 to
285 ppm in the diet generally for 2 years
or more. Although  liver pathology was
generally associated with exposure to the
chlorinated insecticides, there was no
firm evidence of a carcinogenic response
in any of the studies.


Conclusion

Aldrin
  For aldrin, the finding of hepatocellular
carcinomas in male B6C3Fi mice and the
findings of the  hepatomas in C3H and
C3HeB/Fe mice, which were later diag-
nosed as hepatocellular carcinomas, in
both males and females, constitute as a
first approximation/'sufficient" evidence
of carcinogenicity in animals, according
to criteria in the EPA's Guidelines for
Carcinogen Risk Assessment. However,
the Guidelines call for a careful analysis
of the nature of the mouse liver-tumor-
only response to ascertain whether there
is a  sufficiently strong  reason to down-
grade the  evidence to "limited." This
downgrading was found not to be appro-
priate since the tumor increases were not
marginal in male  B6C3Fi mice  (which
have a  high spontaneous incidence of
liver tumors), there was a dose-related
increase in the proportion of tumors that
were malignant, and the  response
occurred in both males and females. In
addition, the response occurs in CaH and
CaHeB/Fe  mouse strains with  a  low
spontaneous incidence of liver tumors.
which is a response not subject to the
downgrading factors.
  One factor which would argue for a
"limited"  classification is that the two
C3H strains are genetically closely related
and the B6C3Fi strain derives half of its
gene  pool from these  strains. However,
the fact that  the epoxide metabolite,
dieldrin, produces liver tumors in seven
different  mouse  strains indicates
strongly that the carcinogenic  effect of
aldrin is not limited to a restricted genetic
range  of  mice. Although in  rats, one
adequate  study with  aldrin and  three
adequate  studies  with dieldrin  were
negative or equivocal,  the multiplicity of
mouse strains responding  is considered
to be  enough for classification of "suf-
ficient" evidence in animals.
  In the absence  of adequate evidence
in humans,  the sufficient animal evi-
dence amounts to an  overall classifica-
tion of B2, or "probable" human carcin-
ogen for aldrin.
  For aldrin, the carcinogenic  potency,
obtained by averaging estimates from
most  sensitive species tested (mice), is
17  per mg/kg/day. The potency  using
the most sensitive sex and strain is 23
per mg/kg/day.  These  are  plausible
upper  bounds  for the  increased cancer
risk from  aldrin, meaning  that the true
risk is not likely to exceed these estimates
and may be lower. These estimates have
been  calculated  using the Agency's
preferred methodology in the absence of
specific physiologic, metabolic, or kinetic
information: the  linearized multistage
model with  doses scaled according to
relative  body surface  area.  Other
assumptions about the dose-response
model or  the interspecies dose adjust-
ment may result in lower estimates. The
molecular potency index for aldrin is 6.2
x 103  per mmol/kg/day.  This places
aldrin in the first (most potent) quartile
of suspect carcinogens ranked by the
CAG.

Dieldrin
  For  dieldrin, the  mouse liver  tumor
response in  seven   mouse  strains
(CaHeB/Fe, C3H, CBA/J, Swiss-Webster,
B6C3Fi, and C57BL/6J) along with the
appearance of pulmonary adenomas and
carcinomas and lymphoid tumors in one
strain (CFi) would justify the preliminary
classification  of dieldrin as  having
"sufficient"  evidence in animals. Based
on the nature  of the response there is
no reason to downgrade the classifica-
tion to  "limited," since the carcinoma
response  is unmistakably strong,  it
occurs in both males and females, and
at both high and low doses. The occur-
rence in  several strains implies that it
is not a genetically isolated finding. The
carcinogenicity studies in rats for dieldrin
were considered negative. However, the
ability of most studies to detect carcin-
ogenicity was compromised due to too
few animals, too high mortality, and too
short duration. Although liver pathology
was  associated with  exposure to the
chlorinated insecticides, there was no
firm evidence of a carcinogenic response.
In the absence of  adequate evidence in
humans, the sufficient animal evidence
amounts to an overall classification of B2
or probable  human  carcinogen for
dieldrin.
  For dieldrin, the carcinogenic potency,
obtained by averaging estimates from the
most sensitive species tested, is 16 per
mg/kg/day. The potency using the most
sensitive sex  and  strain, is 55  per mg/
kg/day. This estimate may, however, be
misleading because 100% of the exposed
animals developed cancer. The second
most sensitive estimate is 28 per mg/
kg/day.  These are plausible upper
bounds for the increased cancer risk from
dieldrin, meaning that the true risk is not
likely to exceed these estimates and may
be lower. These  estimates have  been
calculated using the Agency's preferred
methodology  in the absence of specific
physiologic, metabolic,  or kinetic infor-
mation: the linearized multistage model
with doses scaled according to relative
body surface area.  Other assumptions
about the dose-response model or the
interspecies dose adjustment may result
in lower estimates. The molecular poten-
cy index for  dieldrin is 6.1  x  103 per
mmol/kg/day. This places dieldrin in the
first  (most  potent) quartile of  suspect
carcinogens ranked by the CAG.
  Although there  is always uncertainty
in extrapolating potency from animals to
humans, our confidence in these animal
potency estimates is relatively high. This
confidence is  based not only on the tight
clustering of aldrin and dieldrin  potency
estimates, but on  other factors as well.
The potency  estimates for  aldrin are
consistent with the potency estimates for
dieldrin, one of its  metabolites. In
addition, aldrin's potency (and dieldrin's
as well) is based  in part on studies of
mice with low-background tumor rates.
A study of female CaHeB/Fe mice, for
example, saw the tumor  incidence
increase from 4% in controls to 85% in
the treated group. Potencies derived from
high-background strains are consistent

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 with  potencies from low-background
 strains. Furthermore, potencies derived
 from mouse liver tumors are consistent
 with potencies derived from  rat tumors
 for two other related substances, chlor-
 dane and  heptachlor epoxide. Thus,
 mouse liver tumors can  provide a basis
 for estimating cancer potency in humans,
 recognizing the  current issues  asso-
 ciated with use of this type of data and
 attendant uncertainties in a risk estimate
 based upon it.
        This Project Summary was prepared by staff of Office of Health and
          Environmental Assessment, Washington, DC 20460.
        Dharm Singh is the EPA Project Officer fsee below).
        The complete report, entitled "Carcinogenicity Assessment of Aldrin and
          Dieldrin." (Order No. PB 88-139 951/AS; Cost: $19.95, subject to change)
          will be available only from:
               National Technical Information Service
               5285 Port Royal Road
               Springfield. VA 22161
               Telephone: 703-487-4650
        The EPA Project Officer can be contacted at:
               Office of Health and Environmental Assessment
               U.S. Environmental Protection Agency
               Washington. DC 20460
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
Official Business
Penalty for Private Use $300
EPA/600/S6-87/006
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