EPA-650/1-74-011
DECEMBER 1974
Environmental Health Effects Research Series
ON THE MALE
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EPA-650/1-74-011
ACTIONS OF PESTICIDES
AND OTHER DRUGS
ON THE MALE
REPRODUCTIVE SYSTEM
by
Dr. John A. Thomas
West Virginia University Medical Center
Morgantown, West Virginia 26506
Grant No. R801650
ROAP No. 21AYL
Program Element No. 1EA078
EPA Project Officer: Dr. William F. Durham
Pesticides and Toxic Substances Effects Laboratory
National Environmental Research Center
Research Triangle Park, North Carolina 27711
Prepared for
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
WASHINGTON, D.C. 20460
December 1974
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This report has been reviewed/by -the Environmental Protection Agency
and approved for publication. Approval does not signify that the
contents necessarily reflect the views and policies of the Agency,
nor does mention of trade names or commercial products constitute
endorsement or recommendation for use.
11
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Page
List of Tables i v
Abstract vi i
Acknowledgements vi i i
SECTIONS
I. Conclusions 1
II. Recommendations 5
III. Introduction 6
IV. Experimental Design. 8
V. Evaluation of Results 12
VI. Discussion of Results 26
VII. References 29
VIII. List of Publications 31
IX. Glossary 33
X. Appendix 34
111
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Number
1. Accumulation of Radioactive Steroid in Mouse
Anterior Prostate Glands Following Oral
Administration of DDT (daily x 10) 14
2. Accumulation of Radioactivity as Various
Hydroxylated Metabolites of Testosterone
Following a 15 Minute Incubation of Liver
Microsomes with 1,2-H^ Testosterone 15
3. Effects of Orally Administered Dieldrin (daily
x 10) on the Metabolism and Accumulation of
1,2-H3 Testosterone by the Mouse Anterior
Prostate Gland 16
4. Effects of Dieldrin (2.5 mg/kg. daily x 10) on the
In. Vitro Metabolism of 1,2-H* Testosterone by
Liver Homogenate Obtained From Male Mice 17
5. In Vitro Metabolism of H^ Testosterone by Mouse
Anterior Prostates 18
6. Effects of Orally Administered 2,4,5 T (daily x 10)
on the Accumulation and Metabolism of H^-
Testosterone by the Mouse Anterior Prostate
Gland 19
7. Effects of 2,4,5 T Pre-treatment (12.5 mg/kg,
daily x 10) Upon In Vitro Hepatic Hydroxylation
of HJ-Testosterone 20
8. Effects of Orally Administered Organochlorines
(daily x 10) on Reproductive Organs and Sex
Accessory Tissue in the Mature Male Mouse 21
9. Effects of Varying Doses of Parathion on Repro-
ductive Organs of Male Mice and on the Five
Minute Accumulation and Metabolism of H^-
Testosterone by Anterior Prostate Gland 22
IV
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Number Page
10. Effects of Parathion (daily x 5 po) on the
In. Vitro Bio transformation of H^-
testosterone by Hepatic Microsomes of the
Mouse 23
11. Effects of Varying Doses of Carbaryl (daily
x 5 po) on Reproductive Organs of Mature Male
Mice 24
12. Effects of Thiophanate and Thiophanate-Methyl
on Organ Weights and on the Five Minute
Accumulation of H^-Testosterone by the Prostate
Gland of Mature Male Mice 25
v
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A-B-S-T-R-A-C-T
DDT administered to male mice caused significant reductions
in the assimilation of androgens by sex accessory organs. DDT
altered the metabolism of testosterone in the prostate gland and
in the liver. DDT-^H administration led to the detection of
significant amounts of tritium in male reproductive organs.
Dieldrin caused significant alterations in the assimilation
and metabolism of testosterone-l,2-3H in sex accessory organs
and in hepatic microsomes of rodents.
The herbicide 2,4,5 trichlorophenoxy acetic acid (2,4,5 T)
was effective in altering the metabolism of androgens in male
reproductive organs, but had little effect upon the hepatic
microsomal metabolism of androgens.
Unlike the organochlorine-type pesticides, carbaryl adminis-
tration failed to alter androgen metabolism. The administration
of carbaryl-l^C led to detectable amounts of radioactivity in
several organs of reproduction including the seminal vesicles,
prostate gland and testes. Low concentrations of labeled carbaryl
and/or its radiometabolites were detected in the seminal plasma
of the male mouse.
Parathion administration did not produce any significant
changes upon the uptake and/or the metabolism of androgens by sex
accessory organs of the mouse.
The fungicide, thiophanate and thiophanate-methyl had no
affect upon spermatogenesis or upon the metabolism of androgens.
This agent did significantly increase the weights of the adrenal
gland.
This report was submitted in fulfillment of Grant Number
R801650 by West Virginia University under the full sponsorship
of the Environmental Protection Agency. Work was completed as
of December 11, 1974.
VI1
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ACKNOWLEDGMENTS
The Principal Investigator wishes to acknowledge the
interest and scientific support of several graduate students
involved in aspects of this study, and in particular, Mr.
Lonnie G. Schein, Ms. Cindy Dieringer and Dr. Tim Smith.
Mr. Schein1s interest in the biological consequences of
pesticides is very noteworthy.
The Principal Investigator also wishes to acknowledge
the close monitoring and scientific expertise of Drs.
W. Durham and R. Barren toward this project. Such an
interaction aided in the accomplishment of the many objec-
tives set forth in the EPA supported project.
Vlll
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SECTION I
CONCLUSIONS
ORGANOCHLORINES
DDT
Orally administered technical grade DDT (12.5, 25 or 50 mg/kg) was
studied in intact male mice and in intact and ovariectomized fe-
male mice. Regardless of the dose of DDT, there was a significant
reduction (P^O.05%) in the prostate gland's ability to assimilate
radioactive testosterone. The mechanism(s) of inhibitory action of
DDT upon male sex accessory glands did not appear to be due to its
reported inherent estrogenicity. Single jx> doses of DDT-% revealed
considerable amounts of radioactivity localized in several male
reproductive organs. The prostate and the testes exhibited signifi-
cant amounts as early as one and two hr post-administration.
Epididymal fat pads retained amounts of radioactive pesticide as
long as twelve days after DDT-% ingestion.
The 10-day oral administration of DDT (25 or 50 mg/kg) led to
significant reductions in the accumulation of 1,2-%- testosterone
and its principal metabolite 1,2-%- 5
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labeled androgens in the mouse anterior prostate gland. The in vivo
metabolism of 1,2-%-testosterone to 3H-dihydrotestosterone (^H-DHT) ,
•^H-androstanediol or -^H-androstenedione by the mouse prostate gland
was lowered by pretreatment with dieldrin. Similarly, the rn vitro
metabolism of l,2-3n-testosterone to these aforementioned radio-
metabolites was reduced by dieldrin at a treatment level of 5 mg/kg
(daily x 5 ^>pj . This highest dose regime also reduced the formation
of the metabolites of testosterone in mouse hepatic microsomes.
Varying concentrations of dieldrin (4 x 10"?, 4 x 10~6, or 2 x 10~5 M)
in vitro effectively decreased the formation of H^-DHT in the mouse
anterior prostate gland and of ^H-androstanediol in the rat ventral
prostate gland.
The oral administration of varying doses (2.5 or 5 mg/kg daily x 10)
of dieldrin led to significant reductions in the accumulation of
radioactive androgen by the mouse prostate gland. This was reflected
not only by a reduction in the total accumulation of tritiated steroid
( % _x) by prostate gland, but also by a decrease in the concentra-
tion of dihydrotestosterone (3H-DHT). The ratio of ^H-T to 3H-DHT
remains, however, similar in treated and control groups. The
doses of dieldrin used had little effect upon sex accessory organ
weights or prostatic fructose concentration. Dieldrin administration
failed to alter gonadal weights. There was no change in the hepatic
formation of radioactive dihydrotestosterone, androstanediol or
androstenedione from 1,2-^H-testosterone in the dieldrin-treated
animals, but dieldrin did produce increases in liver androgen hydroxy-
lase activity. While the amounts of dieldrin used in these studies
were quite high, the results, nevertheless, indicate that this
organochloride could represent a potential deleterious chemical from
the standpoint of the male reproductive system.
2,4,5-T
High doses of 2,4,5 trichlorophenoxy acetic acid (2,4,5 T) (6.25,
12.5, or 25 mg/kg daily x 10) were administered orally to mature
male mice. The herbicide 2,4,5 T significantly reduced the assimi-
lation of radioactive androgen by the prostate gland (P<£0.05). This
was reflected not only in a reduction in the total accumulation of
1,2-3H-testosterone by the prostate gland, but also in a decrease
in the levels of its labeled metabolites. There was no change in
the hepatic formation of either polar metabolites or of dihydro-
testosterone, androstanediol, or androstenedione from -%-T in
-2-
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the 2,4,5-T treated mice. This pesticide had little or no effect
on the weight responses of either the testes or the sex accessory
organs. Prostate gland fructose, a chemical indicator of androgenic
activity, was not altered by 2,4,5-T treatment.
ORGANOPHOSPHATES
Parathion
The daily oral administration of parathion (1.3, 2.6 or 5.3 mg/kg)
had no effect upon the uptake and subsequent metabolism of 1,2-3R-
testosterone by the mouse prostate gland. These dose schedules also
had no effect upon the ability of the prostate gland to biotransform
labeled testosterone to its principal radiometabolites. Hepatic
microsomes obtained from mice previously treated with parathion
failed to reveal any alterations in their capacity to hydroxylate
1,2-^H-testosterone. Since the amounts of 6fl, 7dt or 16A-3R hydroxy-
testosterone were unchanged by parathion treatment, androgen hydroxy-
lase activity was not affected by this organophosphate pesticide.
Although these polar radiometabolites remained unchanged by parathion
pretreatment, ^n-androstanediol content was enhanced. ^H-dihydro-
testosterone (^H-DHT) and ^H-androstenedione formation from l,2-3n-
testosterone by hepatic microsomes were not affected by parathion
pretreatment. Various in vitro concentrations of parathion (4 x 10"',
4 x 10~6 or 2 x 10-5 M) significantly reduced the formation of ^n-DHT
in the mouse prostate gland, but not in the rat prostate gland. In
the rat prostate, ^n-androstenedione concentrations were significantly
increased by parathion in vitro.
CARBAMATES
Carbaryl
Carbaryl (l-naphthyl-N-methylcarbamate), a carbamate pesticide, was
investigated with regard to its effects on certain metabolic aspects
of the male mouse reproductive system. A 5-day period of treatment
with carbaryl had little effect on the ability of the anterior pros-
tate gland to biotransform 1,2-^H-testosterone to its major nonpolar
-3-
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radiometabolites in vitro. This 5-day treatment period resulted in
alterations in hepatic 3H-hydroxytestosterone derivatives; carbaryl
significantly increased 16 d[-^H-hydroxytestosterone. The in vitro
incubation of carbaryl with prostate tissue and 1,2-%-testosterone
resulted in a stimulation of ^H-dihydrotestosterone formation,
suggesting a direct action of carbaryl on prostatic steroidogenesis.
A 5-day treatment period using varying doses of carbaryl (8.5, 17 or
34 mg/kg daily, P.O.) failed to alter significantly the ability of the
prostate gland to assimilate 1,2-3R-testosterone. Neither testicular
nor sex accessory gland weights were affected by these regimens of
carbaryl. A single oral dose of ^C-carbaryl (24 )iCi/kg equivalent
to 0.9 mg/kg) led to detectable amounts of radioactivity in several
organs or reproduction including the prostate gland, seminal vesicles
and testes. Very low concentrations of labeled carbaryl and/or its
radiometabolites were detected in the epididymal fat pads and in
the seminal plasma.
FUNGICIDES
Thiophanate
The daily oral administration of thiophanate (295 mg/kg) or thiophanate-
methyl (195 mg/kg) had no affect upon spermatogenesis or the ability
of the prostate gland to assimilate l,2-3n-testosterone. Little
change occurred in the weights of the sex accessory organs, though
there were significant increases in the weight of the prostate gland.
The weights of the adrenal glands also increased significantly in the
treated mice.
-4.
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SECTION II
RECOMMENDATIONS
The current series of studies encompassed the action of
single pesticides with male reproductive systems.
It was found that the organochlorine pesticides are the
principal class of pesticides that exert significant changes
upon male reproductive systems. Although each of the repre-
sentative compounds studied from this class of pesticides have
the same basic effects upon the male reproductive system, the
mechanism of their actions seems to be different. The present
studies were not specifically designed to elucidate the mechan-
ism of action of the various compounds. Future studies should
certainly be devoted to examining some of these mechanisms.
Another recommendation to be considered is pesticide
interactions. Pesticide synergism or interaction is an area
in which remarkably little work has been done. Pesticide
interactions are important since public exposure to pesticides
is rarely to a single compound. Preliminary experiments in-
volving pesticide interaction have been performed within this
laboratory and have confirmed the interaction of more than
one pesticide upon the endocrine system.
Limited parathion-dieldrin interaction studies have shown
that the in vitro metabolism of Testosterone-H^ to its princi-
pal metabolites by prostate glands from male mice were increased
significantly by simultaneous or successive pre-treatment with
dieldrin and parathion. Similar preliminary results were ob-
tained when hepatic microsomal protein and cytochrome P-450
concentrations were investigated. Such studies require more
extensive investigation.
A final recommendation is that toxicological and pharma-
cological examinations of pesticide products newly marketed
should be expanded to assess their affects upon mammalian
reproductive systems. It is necessary to obtain complete
toxicological profiles on commercially available pesticides,
singly or in combination, and to what extent the actions can
alter endocrine function.
-5-
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INTRODUCTION
Much literature exists pertaining to the effects of pesti-
cides on both the vertebrate and the invertebrate animals. But
little specific attention has been devoted to the actions of these
compounds upon the endocrine system, and in particular their
effects upon the male reproductive system. There is growing evi-
dence that some of these compounds can adversely affect hormonal
balance in mammals. Some of these pesticide-induced changes in
hormonal balance can be mediated by alterations in hepatic micro-
somal enzyme systems (Hart and Fouts, 1963; Conney et al, 1967;
Welch, £t al, 1967; Abernathy e_t al, 1971). There also have been
documented sex differences in the metabolism of pesticides (Wong
and Terriere, 1965). Other pesticide-induced changes in hormonal
balance can be ascribed to their direct actions upon such organs
as the testes and the ovaries. They also produce changes in
steroid-dependent organs such as the uterus and the prostate.
Reports have indicated that the organochlorines (e.g. DDT,
Dieldrin, 2,4,5 T) can adversely affect the male reproductive
function. Investigation with dieldrin suggests that it can
affect androgen target organs such as the prostate gland (Thomas
ejt al, 1973; Thomas and Schein, in press; Thomas and Lloyd, 1973).
Wakeling e_t al (1972) showed that dieldrin significantly reduced
both nuclear and cytoplasmic in vitro binding of dihydrotestosterone
(DHT) to a prostatic protein fraction.
Relatively few studies have been concerned with the organo-
phosphate pesticides. Swan e_t al (1958) reported that the acute
toxicity of parathion was related to the sex of the animal and
the sexual maturity gave added protection against the toxicity
of parathion.
The carbamates, more specifically carbaryl, reportedly has
an affect upon the adenohypophysial activity by increasing the
synthesis and release of gonadotropin (Schtenberg & Rybakova,
1968). Little attention has been devoted to the actions of
the carbamates on androgen metabolizing tissues such as the
prostate or the liver. Carbaryl, like parathion, has been shown
during this series of experiments to have no demonstrable effect
upon the male reproductive system (Dieringer and Thomas, 1974).
-6-
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Human male reproductive function also appears to be affected
by exposure to pesticides as evidenced by the fact that impotence
was reported by four out of five farm workers following exposure
to several pesticides and herbicides (Espir et al, 1970).
The findings discussed in this report reveal the effects of
the various pesticides upon the male sex accessory organs and
upon the hepatic microsomal enzyme system involved in androgen
metabolism.
More specifically these studies were designed to study the
effects of pesticide exposure upon the in vivo and the in vitro
assimilation of H-^-testosterone by the prostate gland. The pros-
tate gland is a very representative organ insofar as male sex
hormone responsiveness. Further, the in vivo and in vitro
metabolism of the radiosteroid by the prostate gland, and upon
the in vitro metabolism of radiosteroid by the hepatic microsomal
enzyme system was clearly examined. The gravimetric responses
of the testes and other sex accessory organs were investigated.
Some studies were concerned with prostate gland fructose concen-
tration since this parameter provides an additional index of
androgenic activity.
The compounds investigated in this study were:
1. Organochlorines - DDT, Dieldrin, 2,4,5-T
2. Organophosphate - Parathion
3. Carbamate - Carbaryl
4. Fungicide - Thiophanate and Thiophanate-Methyl
(per request of EPA)
-7-
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EXPERIMENTAL DESIGN
Animal Techniques
Mature male Swiss-Webster mice weighing between 30 and 40 g
were used in these experiments. All animals were obtained from
Hilltop Lab Animals located in Scottdale, Pennsylvania. The
animals were fed a standard diet of laboratory chow and water ad
libitum. Some limited experiments used dogs.
Pesticide Dose Regimen
The pesticides and herbicides used for the in vivo studies were
dissolved in corn oil and administered by gastric incubation
in a volume of approximately 0.1 ml. Control animals received a
similar volume of corn oil. The fungicides (viz. thiophanate and
thiophanate-methyl) were suspended in a 1% Methocel© solution.
The pesticides were ordinarily administered over a period of 5 or
10 days (e.g. 1.25, 2.50 or 5.00 mg/kg dieldrin per day), and the
animals were sacrificed 24 hours after the administration of the
final dose. A no-effect level insofar as the weights of endocrine
organs was frequently used to study subtle biochemical changes.
The in vitro studies examined the effects of varying concentrations
of the compound being studied (4 x 10~7, 4 x 10~°, and 2 x 10~5 M)
upon H^-testosterone metabolism utilizing the prostate and the
livers of the experimental animals. The pesticide solution was
added (20 jil) directly into the incubating medium. Control groups
received a 20 jil volume of corn oil.
Radiosteroid Accumulation By The Prostate In Pesticide-Treated Mice
Mouse anterior prostate glands (also called coagulating glands)
were rapidly excised and frozen in liquid nitrogen five
-8-
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minutes after a single intraperitoneal injection of 1,2-%
testosterone (100 pCi/kg or 10 jig/kg) (spec. act. 43.5 jiCi/mM)
(New England Nuclear Co.). This 5 minute in vivo uptake of radio-
active testosterone was studied 24 hours after administration of
the final pesticide dose. In normal mice this uptake interval
reveals significant androgen assimilation.
After sacrifice the prostate glands were frozen, weighed,
and homogenized in glass distilled water. The homogenates were
rinsed 3 times with chloroform-ether (2:1) (efficiency of ex-
traction was approximately 70%). The samples were subsequently
shaken vigorously for 30 minutes and then allowed to stand over-
night at 4°C in order to effect partition of the organic and
aqueous layers. Aliquots of the organic phase of the samples
were removed, evaporated to dryness under a stream of nitrogen,
and resuspended in chloroform. A suitable scintillation cock-
tail was added to aliquots of the resuspension for subsequent
measurement of radioactivity. Such measurements were provided
an index of total radiosteroid (e.g. H^-testosterone + H^-
DHT, etc.).
Twenty microliter aliquots of the resuspended prostate
homogenates were spotted on Silica-Gel-G (Eastman Co.) thin
layer chromatography (TLC) plates to examine the content of
specific radiometabolites of H-*-testosterone. This TLC technique
utilized a solvent system consisting of chloroform-ether (7:3).
Visualization of radiosteroid metabolites was accomplished by
iodine vapour. This particular TLC system effectively separates
androstanediol (Rf=.507), androstenedione (Rf=.801), dihydro-
testosterone (Rf=.680), and testosterone (Rf=.591). After
visualization, the spots were scraped from the plates into scin-
tillation vials for subsequent assessment of their radioactivity.
Some experiments isolated the polar metabolites of testos-
terone. Prostatic and hepatic androgen hydroxylase activity was
examined following pesticide administration. Several hydroxy-
testosterone derivatives were separated (6p, 7
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Preparation Of Hepatic Microsomes From Mouse & Rat Livers
Liver samples (2 g) were rapidly excised and homogenized
(Polytron 09) in 2 volumes of ice cold 1.5% KC1. The homogenate
was centrifuged (9000 x g) for 25 minutes at 4°C. The super-
natant was decanted and subsequently re-centrifuged at 100,000
x g for 60 minutes at 4°C to guild a microsomal pellet. The
microsomal pellet was resuspended in 3 volumes of ice cold phos-
phate buffer (pH 7.4) and adjusted so that 333 mg of fresh
liver was equivalent to 20 mg of microsomal protein/ml. Proteins
were measured using the Lowry technique. Aliquots (0.5 mis) of
this adjusted microsomal fraction were used for in vitro incubation
with 1,2-H3-testosterone (3.4 x 10'8M) .
In Vitro Incubations
Hepatic microsomes or lobes of the prostate gland were
incubated for 60 minutes at 37°C (or for 15 minutes in some of the
experiments) in a Krebs-Ringer bicarbonate buffer (pH 7.4). This
buffer system also contained 4mM NADP, 5mM glucose-6-phosphate,
5mM MgCl2, 4>iM nicotinamide, 5 I.U. glucose-6-phosphate
dehydrogenase, and 1,2-H^-testosterone. The total volume was
1 ml. Tissues were incubated aerobically in a Dubnoff Metabolic
shaker. Reactions were terminated by addition of 2 ml of ice
cold diethyl ether. The tissues (prostate) were subsequently
removed from the incubation vials and frozen in liquid nitrogen.
Tissue and/or medium radiosteroids were extracted, separated
on TLC plates, and counted in a liquid scintillation system.
Radioactive Pesticide (And/Or Its Radiometabolites) Distribution
Studies
The distribution of radioactivity following -^C-carbaryl
(24 jiCi/kg or 0.9 mg/kg) (Mallinckrodt Radiochemicals) (spec.
act. 5.2 mCi/m mol.) or 3H-DDT (10 jtCi/kg) (New England Nuclear)
(spec. act. 51 Ci/mM) was examined following the single injec-
tion of either radioactive pesticide. Mice were sacrificed at
varying intervals after the administration of the radioactive
pesticide and various tissues (e.g. testes, prostate glands,
epidydimal fat, brain, etc.) were removed, weighed, digested in
solubilizer (NCS) and counted in a Packard liquid scintillation
spectrometer. No attempt was made to chemically identify the
radioactivity.
-10-
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Prostate Gland Fructose Determination
The anterior lobes of the mouse prostate glands were removed
from mice killed by cervical dislocation 24 hours after the final
dose of the pesticide treatment period, blotted, and weighed.
Each gland was homogenized (47o W/V) , precipitated with heavy
metals (i.e. barium hydroxide and zinc sulfate), and the super-
natant was examined for free fructose (Thomas e_t al, 1968).
Fructose concentrations were expressed as mg/100 gm wet weight
of the tissue.
Histological Sections
Testes were removed and were fixed in 57o formalin. They
were sectioned (10 )i) and stained with hematoxylin and eosin.
Statistics
Experimental and control groups were chosen randomly.
Age and weights were variables that were kept constant among the
groups. Usually a minimum of six animals were used per group.
Tissue radioactivity DPM/mg or organ weights (viz, testes,
prostate, etc.) obtained from appropriate controls were compared
with the appropriate pesticide treated groups using either
Student's Jt-test or one-way analysis of variance (Snedecor, 1956)
depending on which was applicable.
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SECTION V
EVALUATION OF RESULTS
Substantial amounts of radiosteroid were found in the mouse
anterior prostate gland five minutes after i.p. injection of
1,2-H^-testosterone (Table 1). However, pretreatment with varying
doses of DDT (10 days) resulted in significant reductions in the
labeled steroid concentration in this gland at the two highest
doses (25 and 50 mg/kg). These reductions were reflected in de-
creases in both H^-testosterone and its principal metabolite H^-DHT.
The effects of a 10-day P.O. regimen of DDT on the hepatic bio-
transformation of H3-testosterone to various metabolites is shown
in Table 2. Substantial reductions were found in the in vitro
formation of 60, 7dL » and 16«Lby liver microsomal preparations
from animals pretreated with the two highest dose levels of DDT.
Pretreatment with dieldrin can significantly decrease the amount
of radiosteroid accumulated by the prostate gland (Table 3).
The highest dose level led to a 4070 reduction in the levels of prostate
gland radiosteroid. Dieldrin failed to produce any marked changes
in the relative proportion of the radiometabolite formed from 1,2-H^-
testosterone; marked changes were found in the absolute amounts of
the specific radiometabolites compared to control values. Dieldrin
enhanced hepatic androgen hydroxylase activity (Table 4). While
the amounts of 60 and 7oC were slightly increased by dieldrin pre-
treatment, the 16«C derivative was significantly increased. This
effect has been found in both the 5 day and the 10 day dieldrin
pre-treatment studies.
Because of these observed effects (i.e. Table 4), it was of interest:
to examine the in vitro accumulation of radiosteroids by the mouse
prostate gland following a 10-day dieldrin pretreatment period
(Table 5). There was a marked increase in the accumulation of
labeled testosterone that took place and the relative proportion
of these androgens was not altered.
Table 6 indicated that a 10-day period of administration of 2,4,5-T
also significantly reduced the assimilation of radioactive steroid
by the mouse prostate gland. The effect of this herbicide on the
relative degree of steroid biotransformation is also shown. No
changes were detected in the relative proportion of the radio-
metabolites of l,2-H3-testosterone, although marked alterations
were found in the absolute levels of radiometabolites.
-12-
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Table 7 shows the effect of a 10-day regimen of 12.5i_mg/kg of
2,4,5-T on the hepatic biotransformation of 1,2-H^-testosterone.
No significant changes were found.
While the assimilation of androgens was reduced by treatment with
the organochlorines, no significant alterations were observed in
the gravimetric responses of either the seminal vesicles or the
prostate glands (Table 8). Similarly, fructose levels in the
prostate glands of DDT and 2,4,5-T were not significantly re-
duced. Prostate gland fructose concentrations in the dieldrin-
treated animals, although not significantly reduced, showed a
trend toward reduction. Testicular weights also remained unchanged,
Table 9 shows that varying doses of parathion had little gravi-
metric effect upon the reproductive organs of the mouse. Neither
the testicular weights nor the prostate gland weights were
affected. The uptake of tritiated testosterone was not affected
by pretreatment with parathion. Similarly, radiometabolites were
not affected by parathion. Regardless of the dose of parathion,
no changes in the pattern of radiosteroids were detected. Para-
thion pretreatment likewise failed to effect any demonstrable
changes in the hydroxylation of 1,2-H^-testosterone by the hepatic
microsomal enzyme system (Table 10).
Carbaryl pretreatment led to slight enhancement in the ability of
the prostate to assimilate injected 1,2-H^-testosterone, but no
statistical elevations were noted (Table 11). Gravimetric res-
ponses of the sex accessory organs remained unaffected. Although
not shown, limited studies have demonstrated that pretreatment
with carbaryl significantly enhanced the 16 dc-hydroxylation of
1,2-H^-testosterone by the hepatic microsomal enzyme system.
The assimilation of labeled testosterone by mice pretreated with
thiophanate or thiophanate-methyl (daily x 5 po) was not affected
(Table 12). This fungicide had no effect upon the gravimetric
responses of sex accessory organs, but it did cause a significant
increase in the weight of the adrenal gland. The histological
sections of the testes revealed that neither thiophanate nor
thiophanate-methyl affected spermatogenesis. No sterile tubules
were observed and the interstitial cells appeared normal.
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TABLE 1
Accumulation of Radioactive Steroid in Mouse Anterior Prostate
Glands Following Oral Administration of DDT (daily x 10).
Daily Dose H^-Testosterone H^ f*-dihydro- Total
(mg/kg) (T^-H)a testosterone Steroid
Control 3594 + 268 2453 + 294 8180 + 816
12.5 3252 + 224 1904 + 179 7298 + 990
25 2305 + 179b 1816 + 161b 5787 + 619b
50 2028 + 116b 1837 ± 152b 5508 + 717b
a Mean + S.E.M. of 6 animals expressed as dpm/
b Significantly Different from control P<0.05
-14-
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TABLE 2
Accumulation of Radioactivity as Various Hydroxylated Metabolites of
Testosterone Following a 15 Minute Incubation of Liver Microsomes With
1,2-H^-Testosterone.
16
DDT (rag/kg x 10 day, po)
Control3 12.5
6B-Hydroxytestos-
terone 850 + 69
835 + 191
776 + 222
25
50
841 + 110 609 + 138b 516 + 146
685 + 86 331 + 53b 253 + 34
845 + 39 300 + 68b 427 + 88
a Mean + SEM of 6 animals expressed as dpm/167 mg equivalents of
liver tissue
Significantly different from control P<0.05
-15-
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TABLE 3
Effects of Orally Administered Dieldrin (daily x 10) on the Accumulation and Metabolism of
1,2-H^-Testosterone by the Mouse Anterior Prostate Gland.a
Total
Dose
(mg/kg)
Control
1.25
2.50
5.00
Androstanediol Testosterone DHT Androst^
7, Total % Change . %Total. %Change
15
14
17
18
--
-30
-24
-35
44
41
41
41
--
-30
-38
-46
7oTotal
30
31
28
27
7o(hange
--
-23
-38
-48
7oTotal
11
14
14
14
pnedione
7o Change
--
1
-15
-25
7o Reduction
of Total
—
-24
-33
-43
-------
TABLE 4
Effects of Dieldrin (2.5 mg/kg, daily x 10) on the In Vitro
Metabolism of 1,2-H^-Testosterone by Liver Homogenates Obtained
From Male Mice.
(dpm/mg) (dpm/mg)
Radiosteroid Control Dieldrin
Testosterone 1000 + 169 866 + 140
6p-hydroxytestosterone 1154 + 61 1346 + 91
7<-hydroxytestosterone 915 + 61 1058 + 80
16Jl-hydroxytestosterone , 3291 + 86 6515 + 95b
a Mean + S.E.M. of 6 animals
Significantly different from control P^O.05
-17-
-------
TABLE 5
In Vitro Metabolism of H^-Testosterone by Mouse Anterior Prostatea
Testosterone DHT Androstenedione
Daily Dose %Total % .Change . % Total .% Change %Total %Change % Total % Change
Control
2.50
23b
19
._
46
34
36
_ _
166
36
38
_~
89
7
8
_ _
123
7o Increg
Total Rf
Steroid
--
108
i Mice were treated po for 10 days with 2.50 mg/kg of dieldrin
00
Values are means for 7 observations
-------
TABLE 6
Effects of Orally Administered 2,4,5 T (daily x 10) on the Accumulation and Metabolism of
H^-Testosterone by the Mouse Anterior Prostate Gland.a
Daily Dose Androstanediol Testosterone Dihydrotestosterone
(mg/kg)
Controlb
6.25
12.50
25.0
%Total
15
15
15
15
7,Change
-4
-19
-37
7oTotal
44
44
42
40
70Change
-13
-22
-40
7oTotal
30
28
28
31
7o Change
-23
-24
-33
Androstenedione Tot*
7o Total
11
13
13
14
7o Change
1
-4
-14
Total
70
Re due .
--
14
19
35
a
Mice were killed 5 minutes after ip injection of HJ-testosterone
Mean of 6 or more animals
c Significantly different from control P*0.05
-------
TABLE 7
Effects of 2,4,5-T Pretreatment (12.5 mg/kg, daily x 10) Upon In Vitro
Hepatic Hydroxylation of Hr-Testosterone.
Radiometabolite Control 2,4,5-T
6f Hydroxytestosterone 1084 + 94a 956 + 199
7 dl 825 + 155 1065 + 286
16J, 3364 + 117 3911 + 811
a Mean + SEM of six samples expressed as DPM/167 mg liver tissue
-20-
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TABLE 8
Effects of Orally Administered Organochlorides (daily x 10) on
Reproductive Organ and Sex Accessory Tissue in the Mature Male Mouse,
(mg/kg)
Treatment
Cpntrol
DDT (50)
2,4,5-T(25)
Dieldrin (5)
Organ Weights (mg/kg body wt.)
Testes Sem. Ves. Prostate
8.1+0.3
7.2 + 0.8
7.3 + 0.3
7.2 + 0.4
2.8 + 0.5
2.6 + 0.1
3.2 + 0.1
2.6 + 0.1
.61 +
.72 +
.56 +
.71 +
.03
.05
.02
.05
Prostatic Fructos
mg/100 gm wet wt.
.19 +
.20 +
.19 +
.14 +
.02
.01
.02
.01
a Mean + SEM of at least 6 organs
Mean + SEM of at least 12 organs
-21-
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TABLE 9
Effects of Varying Doses of Parathion on Reproductive Organs of Male Mice and on the Five
Minute Accumulation and Metabolism of H3-Testosterone by the Anterior Prostate Gland of the
Mous e.
Total Prostate
Daily Dose Testes wt. Prostate wt. Radioactive
(mg/kg) (mg) (mg) (dpm/mg) H3-T H3-DHT H3-di
1
S3
•0
1
0
1.3
2.6
5.3
268
249
269
268
± 8a
± 8
± 12
+ 16
31
32
34
32
+ 1
± 2
+ 1
+ 1
495
532
755
692
+ 96
± 78
+ 33
+ 84
239
260
471
421
+ 37b
+ 38
± 51
+ 87
85 + 21
83 + 14
87+6
86+6
14 +
19 + 0.
12 +
14 +
ct
Mean + SEM of 6 or more animals
dpm/mg
c Testesterone-H3
^ Dihydrotestosterone-H3
e Androstenediol-H3
-------
TABLE 10
Effects of Parathion (daily x 5 po) on the In Vitro Bio transformation
of H -Testosterone by Hepatic Microsome of the Mouse. a
Polar Metabolites
(mg/kg)
Daily Dose
,0
1.3
•
2.6
5.3
Microsome Incubated for 60 minutes at 37°C
Mean + SEM of six animals
16* Hg
163 + 12b
144 + 8
137 + 13
152 + 9
74 Kg
57 + 3
63+6
67 + 6
57 + 5
6P Hg
34 + 3
36+2
30+2
40 + 3
-23-
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TABLE 11
Effects of Varying Doses of Carbaryl (daily x 5, po) on Reproductive
Organs of Mature Male Mice
Daily Dose
(mg/kg)
Testes Weight
(mg/g)
Prostate Weight
(mg/g)
Prostate
Radioactivity
(dpm/tng) a
0
8.5
17
34
7.0 + 0.5C
7.1 + 0.2
7.2 + 0.4
7.8 + 0.2
0.70 + 0.09
0.67 + 0.08
0.80 + 0.11
0.72 + 0.11
235 + 65
442 + 90
406 + 91
367 + 88
-24-
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TABLE 12
Effects of Thiophanate and Thiophanate-Methyl on Organ Weights and on the five Minute
Assimilation of H^-Testosterone by the Prostate Gland of Mature Male Mice.
Group
Control
Thiophanate (275 mg/kg)
i Thiophanate-Methyl
Body Weight
(g)
37.0 + 1.2a
35.2 + 1.2
Testes
(mg)
236 + 10
268 + 10
Prostate
(mg)
26 + 2
31 + 3
Sem. Ves.
(mg)
157 + 16
133 + 10
Adrenal
(mg)
9.3 + 0.8
13.6 + l.lc
Total
Radioac
304 +
311 +
(192 mg/kg) 35.4 + 0.8 271 + 11 33 + lc 146 + 5 18.0 + 0.6C 294 +
a Mean + SEM of eight animals
b DPM/mg
c Significantly different from control (P<0.05)
-------
DISCUSSION OF RESULTS
These studies demonstrate that DDT (12.5, 25 and 50 mg/kg)
can result in a decrease in the five minutes in vivo accumulation
of labeled ^H-testosterone and its principal metabolite ^H-DHT
by male organs of reproduction. The ratio of ^n-testosterone to
3R-DHT, however, did not change significantly, suggesting that
the uptake, and not the metabolism of %-testosterone was altered
by pre-treatment with DDT. Several organochlorine pesticides
including DDT have a well documented affect on hepatic steroid
hydroxylase enzyme systems (Conney, et al, 1967; Hart and Fouts,
1965). Because of reported species differences with regard to
the effect of DDT on liver hydroxylating enzymes, it seemed
important to examine the action of this pesticide upon both
hepatic and prostatic steroid metabolizing enzymes. A ten-day
oral pre-treatment of mice with DDT which resulted in a decrease
in accumulation of radioactive steroid by the prostate gland
caused substantial reductions in the ability of the liver to
form radioactive polar metabolites. Since polar metabolites of
testosterone catabolism are more readily excreted, it would not
be surprising that such changes could contribute to altered endo-
crine states in DDT-treated animals. It is possible that DDT
transiently increased circulating levels of hormone available
to the prostate gland thus rendering the prostatic receptor
sites less available for the injected H^-testosterone. A
similar mechanism has been suggested by Blend and Visek (1972)
to explain their observations that dieldrin decreased the
effectiveness of chlormadinone to reduce canine prostatic se-
cretions. It should be noted however that if endogenous con-
centrations of androgen were elevated by prior DDT treatment,
the elevations were not great enough and/or not high enough to
alter sex accessory weights in prostate gland fructose concen-
trations. Future studies should investigate endogenous levels
of circulating male sex hormones. Wakeling ejt al (1973) has
reported that DDT, as well as dieldrin, inhibit the in vitro
binding of ^H-DHT to a specific receptor. Distribution studies
from this laboratory using HP- revealed elevated levels of
radioactivity localized in male gonads, anterior lobes of the
prostate gland and seminal plasma of mice 2 hours after i.p.
administration. This indicates that DDT has the potential to
be transmitted to the female with the concomitant possibility of
genetic alterations occurring.
The dieldrin studies demonstrated that a 5 or 10 day pre-
treatment causes a marked reduction in the accumulation of
-26-
-------
1,2-H3-testosterone by the mouse prostate gland without affecting
the relative proportions of the radiometabolites as compared to
the control animals. This would suggest that the decrease in the
accumulation of radiosteroids by the prostate gland was the result
of altered uptake rather than a change in the bioconversion of
testosterone. Such findings warranted studying the effects of
dieldrin pretreatment upon the hepatic microsomal hydroxylation
system. Dieldrin pretreatment resulted in a significant increase
in the 16 -hydroxylation of testosterone. Since these polar
metabolites of testosterone are more readily excreted, it would
be expected that there would be an alteration in the endocrine
status of the dieldrin pretreated animal. It is possible that
dieldrin decreased the amount of circulating hormones available
to the prostate gland. A reduction of circulating androgens is
suggested by the trend toward a decrease in prostate gland fruc-
tose concentration. The increased incorporation of labeled
steroid observed in the prostate glands of dieldrin pretreated
animals incubated in vitro with 1,2-H^-testosterone further
supports the idea of reduced circulating androgens.
Studies with 2,4,5-T. like that of DDT and dieldrin, also revealed
a reduction of 1,2-H^-testosterone assimilation by the mouse pros-
tate gland. 2,4,5-T did not affect the relative proportions of
the radiometabolites. Although pretreatment with 2,4,5-T caused
a significant reduction of the accumulation of l,2-H3-testosterone,
no affect could be detected on the hepatic formation of labeled
hydroxylated androgens. Endogenous levels of blood androgen
levels appear not to be affected by the administration of 2,4,5-T
since there were no alterations in either sex accessory weights
or prostatic fructose concentrations. These results indicate,
therefore, that 2,4,5-T may exert a direct affect upon the prostate
gland such as that reported to exist for dieldrin and DDT. Only
additional studies will reveal if 2,4,5-T inhibits androgen binding
in target tissues.
The organophosphate studies indicated that unlike the organochlorine,
parathion did not produce significant alterations in either uptake
or biotransformation of 1,2-R3-testosterone. No significant
changes were noted in the formation of polar metabolites by hepatic
microsomal fraction from parathion-pretreated mice, although
Kuntzman et al (1966) showed that several of the organophosphate
pesticides produce significant reductions in the formation of polar
metabolites from testosterone by rat liver fractions.
-27-
-------
The carbamates, like the organophosphates, did not appear to
exert any demonstrable effect upon the male reproductive system.
Early studies by Ware and Good (1967) observed no detectable
differences in reproductive success of mice when pretreated
with different carbamate pesticides. Guthrie e_t a_l (1971) re-
ported similar findings. Carbaryl was found to have no effect
on the assimilation of radioactive testosterone by the mouse
prostate gland or upon the in vivo metabolism of testosterone by
the prostate gland (Thomas e_t al, 1973). However, carbaryl
does seem to have the ability to enhance the production of 16
hydroxylated testosterone metabolites in liver microsome frac-
tion. Although carbaryl can increase 16 -hydroxylase activity
of the liver, circulating androgen levels did not seem to be
affected as evidenced by the lack of significant changes in the
prostate gland weights and the gonadal weights.
The thiophanate and thiophanate-methyl assays were ancillary to
the objectives and primary goals of the present series of experi-
ments. Extensive studies could not be performed with these com-
pounds due to the limited available quantities of these fungi-
cides. Neither fungicide affected spermatogenesis. The ability
of the prostate to assimilate HP-testosterone was not affected
by either thiophanate or thiophanate-methyl. There were no
important changes in absolute sex accessory organ weights,
although thiophanate-methyl caused significant increases in
prostate gland weight. This may be rated to the significant
increases in adrenal weights. It is possible that these were
stress-producing agents.
In summary, therefore, this series of studies has demonstrated
that the carbamates alone or the organophosphates alone exerted
little effect upon the endocrine balance in the male reproductive
system of the mouse. However, all three organochlorine compounds
(viz. DDT, dieldrin and 2,4,5-T) employed in this series of
investigation substantially reduced the uptake of radioactive
testosterone by androgen dependent organs. No common site of
action is as yet clearly evident. Changes in hepatic steroid
hydroxylase enzyme activity may, in part, explain the actions of
DDT and dieldrin. Further, although all three compounds may be
exerting an inhibitory action at the level of endocrine target
organs (e.g. testosterone or DHT-binding and/or transport), it
seems unlikely that reported inherent estrogenicity (in the case
of technical grade DDT) accounts for their actions. Further
experiments may fully resolve their molecular mechanism(s) upon
the reproductive system.
-28-
-------
SECTION VII
REFERENCES
AbernatHy, C.O., Hodgson, E. and Guthrie, F. Structure-Activity
Relationship on the Induction of Hepatic Enzyme in the
Mouse by l,l,l-Trichloro-l,2-Bis-(p-Chlorophenyl)-Ethane
(DDT) Analogs. Biochem. Pharmacol. 20:2385, 1971.
Blend, M.J. and Visek, W.J. Effects of Low Concentrations of
Dieldrin and Chlormadinone on Canine Prostatic Fluid.
Toxicol. Appl. Pharmacol. 23:344-348, 1972.
Conney, A.H., Welch, R.M., Knutzman, R. and Burns, J.J. Effect
of Pesticides on Drug and Steroid Metabolism. Clin.
Pharmacol. and Ther. 8:2-10, 1967.
Dieringer, C.S. and Thomas, J.A. Effects of Carbaryl on the
Metabolism of Androgens in the Prostate and Liver of the
Mouse. Environ. Res. 7:381-386, 1974.
Espir, M.L.E., Hall, J.W., Shirreffs, J.G. and Stevens, D.L.
Impotence in Farm Workers Using Toxic Chemicals. Brit.
Med. J. 1:423-425, 1970.
Hart, L.C. and Fouts, J.R. Effects of Acute and Chronic DDT
Administration on Hepatic Microsomal Drug Metabolism in
the Rat. Proc. Soc. Exp. Biol. Med. 114:388-392, 1963.
Guthrie, F.E., Monroe, R.J. and Abernathy, C.O. Response of
Laboratory Mouse to Selection for Resistance to
Insecticides. Tox. Appl. Pharmacol. 18:92-101, 1971.
Kuntzman, R., Welch, R. and Conney, A.H. Factors Influencing
Steroid Hydroxylases in Liver Microsomes. Advan. Enzyme
Reg. 4:149-160, 1966.
Schein, L.G. and Thomas, J.A. Effects of Dieldrin on the Uptake
and Metabolism of Testosterone-l,2-3H by Rodent Sex
Accessory Organs. Environ. Res. (in press).
Schtenberg, A.L. and Rybakova, M. Effects of Carbaryl on the
Neuroendocrine System of Rats. Food Cosmet. Tox. 6:
461-467, 1968.
-29-
-------
Snedecor, G.W. Statistical Methods. Iowa State College Press,
Ames, Iowa, 5th edition, 1965. 532 p.
Swann, H.E., Woodson, G.S. and Ballard, T.H. The Acute Toxicity
of Intramuscular Parathion in Rats and the Relation of
Weight, Sex and Sex Hormones to this Toxicity. Amer.
Ind. Hyg. Ass. J. 19:190-195, 1958.
Thomas, J.A., Dieringer, C.S. and Schein, L.G. Effects of Carbaryl
on Mouse Organs of Reproduction. Tox. Appl. Pharmacol.
28:142-145, 1974.
Thomas, J.A. and Lloyd, J.W. Organochloride Pesticides and Sex
Accessory Organs of Reproduction. In "Pesticides and
the Environment: A Continuing Controversy," pp. 43-51.
Intercontinental Medical Book Corporation, New York.
Thomas, J.A., Lloyd, J.W., Smith, M.T., Mawhinney, M.G. and Smith,
C.T. Effect of Dieldrin on the Accumulation and Biotrans-
formation of Radioactive Testosterone by the Mouse Prostate
Gland. Tox. Appl. Pharmacol. 26:523-531, 1973.
Thomas, J.A., Mawhinney, M.G. and Mason, W. Sex Accessory Fructose:
An Evaluation of Biochemical Techniques. Proc. Soc. Exp.
Biol. Med. 127:930-934, 1968.
Thomas, J.A. and Schein, L.G. Effect of Parathion on the Uptake
and Metabolism of Androgens in Rodent Sex Accessory
Organs. Tox. Appl. Pharmacol. (in press).
Wakeling, A.E., Schmidt, T.J. and Visek, W.J. Evidence For
Influence of Dieldrin on Binding of 5-dihydrotestosterone
in Rat Ventral Prostate. Fed. Proc., 31:725, 1972.
Ware, G.W. and Good, E.E. Effects of Insecticides on Reproduction
in the Laboratory Mouse. Ill Tranid R and G.C-9160. J.
Econ. Entomol. 50:530-532, 1967.
Welch, R.M., Levin, W. and Conney, A.H. Insecticide Inhibition
and Stimulation of Steroid Hydroxylases in Rat Liver.
J. Pharmacol. Exp. Ther. 155:167-173, 1967.
Wong, D.T. and Terriere, L.C. Epoxidation of Aldrin, Isodrin,
and Heptachlor by Rat Liver Microsomes. Biochem. Pharmacol.
14:375-377, 1965.
-30-
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SECTION VIII
LIST OF PUBLICATIONS
Smith, M.T., J.A. Thomas, C.G. Smith, M.G. Mawhinney and J.W. Lloyd.
Effects of DDT on Radioactive Uptake from Testosterone-l,2-3R
by Mouse Prostate Glands. Toxicology and Applied Pharmacology.
23 (1):159-164, September 1972.
Smith, C.G., J.A. Thomas, M.G. Mawhinney and J.W. Lloyd. Effect
of Testosterone (T) or Dihydrotestosterone (DHT) on the In
Vitro Synthesis of Labeled Cyclic Adenosine Nucleotide (cAMP-H3)
by Sex Accessory Organs of Reproduction. Federation Proceedings.
31 (2):462, 1972 (abstract).
Smith, M.T., J.A. Thomas, C.G. Smith, M.G. Mawhinney and J.J. McPhillips,
Localization of DDT-^H in Male Reproductive Organs and its
Actions Upon Androgenic Function. Proceedings Soc. Tox.,
March 1972 (abstract).
Thomas, J.A., M.T. Smith, M.G. Mawhinney, C.G. Smith and J.J.
McPhillips. Actions of Dieldrin on the Uptake of Testosterone-
1,2-H3 by Prostate Glands of the Male Mouse. Proceedings Soc.
Tox. March 1972. (abstract).
Lloyd, J.W., J.A. Thomas, M.G. Mawhinney and C.S. Dieringer.
Effects of 2,4,5-T and other Organochlorides on the Metabolism
of (1,2-3R) Testosterone by Male Organs of Reproduction.
Proc. Soc. Tox. March 1973 (abstract).
Lloyd, J.W., J.A. Thomas and M.G. Mawhinney. 2,4,5-T and the
Metabolism of Testosterone-l,2-3H2 by Mouse Prostate Glands.
Arch. Environ. Health. 26:217-220, April 1973.
Thomas, J.A., L.G. Schein, and C. Dieringer. Distribution of Radio-
activity in Male Reproductive Organs After the Administration of
Either Carbaryl-l^C or DDT-3H. The Pharmacologist. 15 (2):227,
August 1973. (abstract).
Thomas, J.A., J.W. Lloyd, M.T. Smith, M.G. Mawhinney and C.G. Smith.
Effect of Dieldrin on the Accumulation and Biotransformation of
Radioactive Testosterone by the Mouse Prostate Gland. Tox. and
Appl. Pharmacol. 26 (4):523-531, December, 1973.
-31-
-------
Thomas, J.A., C.S. Dieringer and L. Schein. Effects of Carbaryl
on Mouse Organs of Reproduction. Tox. and Appl. Pharmacol.
28 (1):142-145, April 1974.
Thomas, J.A. and M.G. Mawhinney. Failure of Parathion to Alter Male
Mouse Reproductive Organ Activity. Proc. Soc. Tox. March 1974.
Thomas, J.A. and L. Schein. Effects of Thiophanate and Thiophanate-
methyl on the Male Reproductive System of the Mouse. Tox. Appl.
Pharmacol. 1974 (in press).
Thomas, J.A. and L. Schein. Effect of Parathion on the Uptake and
Metabolism of Androgens in Rodent Sex Accessory Organs. Tox.
Appl. Pharmacol. 29:53-59, 1974.
Dieringer, C.S. and J.A. Thomas. Effects of Carbaryl on the
Metabolism of Androgens in the Prostate and Liver of the Mouse.
Environ. Res. 7:381-386, 1974.
Lloyd, J.W., J.A. Thomas and M.G. Mawhinney. Prostatic and Hepatic
Testosterone-l,2-3R Metabolism as Affected by DDT Pre-Treatment
in the Mouse. Tox. Appl. Pharmacol. 28 (2):248-252, May 1974.
Schein, L.G. and J.A. Thomas. Effects of Dieldrin on the Uptake and
Metabolism of Testosterone-1,2-% by Rodent Sex Accessory
Organs. Environ. Res. 1974 (in press).
Thomas, J.A., L.G. Schein, H.D. Colby, and W.J. Canady. Interactions
of Dieldrin and Parathion on Androgen Metabolism in the Prostate
and on Hepatic P-450 Activity in Mice. The Pharmacologist. 16
(2):219, 1974 (abstract).
BOOKS:
Thomas, J.A. and J.W. Lloyd. Organochloride Pesticides and Sex
Accessory Organs of Reproduction. In: Pesticides and the Environment:
A Continuing Controversy, Deichmann, W.B. (ed.). New York,
Intercontinental Medical Book Corporation, 1973. p. 43-51.
Thomas, J.A. Effects of Pesticides on Reproduction. In: Molecular
Mechanisms of Gonadal Hormone Action, Thomas, J.A. and Singhal,
R.L. (eds.). Baltimore, University Park Press, 1974.
-32-
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R801650
SECTION IX
GLOSSARY
DDT - (1,1-bis Cp-chlorophenyll -2,2,2-trichloroethane)
Dieldrin - (l,2,3,4,10,10-hexachloro-6,7-epoxy-l,4,4a,5,6,7,8,8a-
octahydro-endo-exo-1,4:5,8-dimethanonaphthalene)
2,4,5-T - (2,4,5-trichlorophenoxy acetic acid)
Parathion - (0,0-diethyl-O- Cp-nitrophenyll ester phosphorothioic acid
Carbaryl - (1-naphthyl-N-methyl carbamate)
Thiophanate - C1,2-bis-(3-ethoxycarbonyl-thiourendo)-benzeneJ
Thiophanate-methyl - Cdimethyl-4,41-o-phenylene-bis-(3
thioallophanate)J
-------
BIBLIOGRAPHIC DATA
SHEET
1. Report No.
EPA-650/1-74-011
3. Recipient's Accession No.
4. Tuif and Subtitle
Actions of Pesticides and Other Drugs on the Male
Reproductive System
5. Report Date
December 1974
6.
7. Author(s)
John A. Thomas
8. Performing Organization Kept.
No.
9. Performing Organization Name and Address
West Virginia Univ. Med. Ctr.
Department Pharmacology
Morgantown, W. Va. 26506
10. Project/Task/Work Unit No.
11. Contract/Grant No.
R801650
12. Sponsoring Organization Name and Address
West Virginia University
Morgantown, WV 26506
13. Type of Report & Period
Covered
9/1/71 - 12/11/74
14.
15. Supplementary Notes
16. Abstracts
DDT or dieldrin or 2,4,5 T administered to male mice caused significant reductions
in the assimilation of androgens by sex accessory organs. DDT or dieldrin altered
the metabolism of testosterone in the prostate gland and in the liver. DDT-3H
administration led to the detection of significant amounts of tritium in male repro-
ductive organs. Unlike the organochlorine-type pesticides, neither carbaryl nor
parathion administration altered androgen metabolism. The administration of carbaxyl-
*4C led to detectable amounts of radioactivity in several organs of reproduction
including the seminal vesicles, prostate gland and testes. The fungicide thiopharate
and thiophanate-methyl had no affect upon spermatogenesis or upon the metabolism
of androgens.
17. Key Words and Document Analysis. 17o. Descriptors
pesticides
male reproductive organs
DDT
Dieldrin
2,4,5 T
testosterone metabolism
prostate gland
hepatic microsomes
parathion
carbaryl
thiophanate
s teroidogene sis
spermatogenesis
17b. Identifiers/Open-Ended Terms
organochlorine pesticides & testosterone metabolism
localization of DDT in reproductive organs
- Dieldrin and DDT effects on testosterone uptake
parathion and androgen metabolism
parathion and androgen metabolism
i7c. COSATI Field/Group Environmental Toxicology; Toxicology (pesticides)
18. Availability Statement
Release unlimited
19. Security Class (This
Report)
UNCLASSIFIED
20. Security Class (This
Page
UNCLASSIFIED
21. No. of Pag'ts
41
22. Price
FORM NTIS-35 (REV. 3-72)
34
USCOMM-DC I4882-P72
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