Effects of Environmental Estrogens to Wildlife/Animals

1. Aquatic wildlife

There is significant accumulated evidence that fish exhibit perturbed development in waters which
receive the effluent from STPs (Hotchkiss et al., 2008). However, it is important to note threshold
concentrations of toxicity by estrogens on fish life. In the U.S. watersheds, human derived estrogens
have a short term predicted-no-effect concentration (PNEC) of 5 ng/L and a long term PNEC of 2 ng/L on
fish (Anderson et al., 2012). Seemingly, EE2 is the most potent hormone with a PNEC of 0.1 ng/L for
aquatic chronic toxicity (Laurenson et al., 2014).

More specifically, several studies demonstrated that elevated concentrations of natural and synthetic
estrogens feminize male fish e.g. reduce testes size (Arnold et al., 2014, Tetreault et al., 2011), affect
reproductive fitness (Rose et al., 2013), lower sperm count, induce the production of vitellogenin (VTG)
(Kidd et al., 2007) and alter other reproductive characteristics (Van Donk et al., 2016). Additionally, EE2
caused a considerable reduction in fish biomass and interrupted the aquatic food chain (Hallgren et al.,
2014). However, a seven-year microsatellite field study revealed that, remarkably, fish can overcome the
effects of EE2 as evidenced by latent increases in fish population levels (Blanchfield et al., 2015). On the
other hand, EE2 does have severe deleterious effects on other forms of aquatic life. For example, in a
recent study, EE2 at 10 ng/L directly affected the heart function of bullfrog tadpoles (Salla et al., 2016).

Despite widespread recognition that municipal wastewaters contain natural and synthetic estrogens,
which interfere with development and reproduction of fishes in freshwaters worldwide, there are
limited data on the extent to which natural populations of fish can recover from exposure to these
compounds. We conducted whole-lake additions of an active component of the birth control pill (17α-
ethynylestradiol; EE2) that resulted in the collapse of the fathead minnow (Pimephales promelas)
population. Here we quantify physiological, population, and genetic characteristics of this population
over the 7 years after EE2 additions stopped to determine if complete recovery was possible. By 3 years
post-treatment, whole-body vitellogenin concentrations in male fathead minnow had returned to
baseline, and testicular abnormalities were absent. In the spring of the fourth year, adult size-frequency
distribution and abundance had returned to pretreatment levels. Microsatellite analyses clearly showed
that postrecovery fish were descendants of the original EE2-treated population. Results from this whole-
lake experiment demonstrate that fish can recover from EE2 exposure at the biochemical through
population levels, although the timelines to do so are long for multigenerational exposures. These
results suggest that wastewater treatment facilities that reduce discharges of estrogens and their
mimics can improve the health of resident fish populations in their receiving environments.

K.A. Kidd, P.J. Blanchfield, K.H. Mills, V.P. Palace, R.E. Evans, J.M. Lazorchak, R.W. Flick
Collapse of a fish population after exposure to a synthetic estrogen
Proc. Nat. Acad. Sci., 104 (2007), pp. 8897-8901

2. Domestic animals
Phytoestrogens (isoflavones, which are structurally and functionally similar to 17β-E2) cause
developmental abnormalities in domestic animals. These effects are manifest morphologically with cows
exhibiting changes in teat length and color of the vulva (Burton and Wells, 2002). Some plants also
contain sufficient concentrations of estrogens to cause reproductive alterations in domestic animals. For
example, sheep grazed on the clover plant, which contains potent levels of phytoestrogens, develop
permanent infertility, so called “clover disease” (Hotchkiss et al., 2008). The effect of estrogens can also
impair vision. Intraocular eye pressure (IOP) varies from species to species in domestic animals but
progesterone can increase the IOP in lions, and estrogen similarly in cats (Shemesh and Shore, 2012).
https://www.sciencedirect.com/science/article/pii/S0160412016304494

3. Some wildlife biologists believe that endocrine-disrupting chemicals are placing many wildlife
populations at risk. Exposure to environmental contaminants has been linked to documented problems
in wildlife that include the following:

· thyroid dysfunction in birds and fish;

· decreased fertility in birds, fish, shellfish, and mammals;
· decreased hatching success in birds, turtles, and fish;
· gross birth deformities in birds, fish, and turtles;
· male fish, birds, and mammals that are feminized;
· female fish, birds, and mammals that are masculinized; and
· compromised immune systems in birds and mammals

Many of these effects are manifest in species living in or near the Great Lakes. These lakes contain many
synthetic chemicals, including byproducts of chemical production (such as PCBs) and pesticides
(including some now restricted or banned, such as DDT). In herring gull embryos and newly hatched
chicks from Lake Ontario, some males have oviducts and gonads resembling ovaries, and the oviductal
system in female birds are developing abnormally. Two- to four-year old salmon show a 100 percent
prevalence of thyroid enlargement. Bird species that feed on Great Lakes fish show behavior changes,
failed reproduction, and early mortality in offspring.
Such findings fuel researchers' growing concern about the potential effects of environmental estrogens.
First, the effects are most often manifest in offspring, not in the exposed parent. Second, the effects of
exposure depend on the timing relative to the organism's stage of development. Third, the effects of
exposure to endocrine-disrupting chemicals may be manifested in entirely different ways in the early
embryo, fetus, and newborn than in organisms exposed only in adulthood. Finally, because the effects of
exposure are often delayed, they may not be fully expressed until the offspring of the exposed adult
reaches maturity or even middle age. If so, the reproductive future for some species may be grim even if
adults continue to breed and juveniles seem healthy.

WRI, (1994) Effects of Environmental Estrogens on Wildlife. Toxics and Health: - The Potential Long-term
Effects of Industrial Activity, p-205-250

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10&cl=CL2.20&d=HASH227fe5d4d2df60d76cf423.7.3&gt=1
4. The paper “Estrogen as an Environmental Pollutant” appeared in the Bulletin of Environmental
Contamination and Toxicology in 1993 (Shore et al. 1993). At the time it was one of the first papers to
suggest that hormones excreted into the environment by humans and animals were present in sufficient
quantities to disrupt the environment.

Although originally an estrogen was defined as a compound that increase the uterine weight in the rats,
in recent times, the ability to bind to the estrogen receptor has become the standard (Dorfman 1962;
USEPA 1998). However, the receptor is also the site for chemicals to exert their anti-estrogenic actions.
In nature there are some 300 natural compounds, and even more anthropogenic compounds, which
have estrogenic or anti-estrogenic activity. These environmental estrogens can be categorized into four
groups: (1) naturally occurring non-steroidal plant estrogens or phytoestrogens; (2) the steroid
estrogens – 17β estradiol and estrone from animal and human sources; (3) the mycotoxins, zearalenone
and zearalenol; (4) synthetic compounds with phenolic groups (Shemesh and Shore 1987; Lintelmann et
al. 2003).

In cows and sheep, the most important function of estrogen is to induce estrus behavior. However, too
much estrogen, hyperestrogenism, can interfere with the function of the reproductive tract. Symptoms
of hyperestrogenism in cattle include nymphomania (constant behavioral estrus), ovarian cysts and
premature udder development (Shore and Bar-El Cohen 2010). Estrogens are also used in animal
husbandry to increase growth in steers. In pigs, uterine prolapse caused by zearalenone is a common
phenomenon.

Phytoestrogens, in particular, are well documented to cause problems of infertility in cattle (Shemesh
and Shore 1987). The function of the phytoestrogen in a legume is to signal the rhizobium to colonize
the nodules. There is some evidence that the compounds may have evolved to be a mechanism to
reduce fertility in foragers as well. A number of decades ago we noticed a rise in the level of coumestrol,
a phytoestrogen, in alfalfa plants to levels associated with reproductive disorders. This was unrelated to
any known trauma which we had previously described (Shemesh et al. 1969). It was found that the rise
in coumestrol was correlated with irrigation by treated sewage water (Shore et al. 1995). When the
treated sewage water was tested, estrone and 17β estradiol were readily detected. Furthermore, using
experimental hothouse and hydroponic experiments, it was shown estrone and 17β estradiol in
concentrations found in the irrigation water promoted vegetative growth (Shore et al. 1992).

In separate line of investigation, it was observed that cattle, which were fed chicken manure silaged with
wheat, developed premature udders. Examination of the chicken manure showed that the amount of
steroidal estrogen ingested (1 mg/9 kg of feed/day) was high enough to explain the phenomenon (Shore
et al. 1988; Shemesh and Shore 1994). Other reproductive disruptions could be attributed to the
testosterone present in the manure (Shemesh and Shore 1994). These steroids were persistent in the
environment as there was no decrease in either the content of testosterone or estrogen following 6
months of silaging (Shore and Shemesh 1993).

L. S. Shore (2016) Estrogen as an Environmental Pollutant, Bulletin of Environmental Contamination and

Toxicology volume 97, pages447–448 https://doi.org/10.1007/s00128-016-1873-9
https://link.springer.com/article/10.1007/s00128-016-1873-9#citeas
5. A study in 1938 found that certain synthetic chemicals could mimic estrogen, and more than 30 years
ago, Rachel Carson’s Silent Spring described how some synthetic chemicals were collecting in and
contaminating water, soil, wildlife and even humans. These chemicals, she warned, were causing severe
health problem (eggshell thinning, cancer, die-offs) in wildlife, especially in species at the top of the food
chain that eat other contaminated animals and accumulate the most chemicals. At high doses, some of
these chemicals can affect an animal’s endocrine system, especially during critical development stages.
For instance, one study showed that fish living near municipal sewage outlets in England had both male
and female sex characteristics and their liver produced vitellogenin, a female egg-yolk protein not
normally found in males. Several different chemicals, especially the alkylphenols, the breakdown
products of chemicals found in detergents and plastics, are suspected of causing the feminizing effects.
Alligators living in Florida’s Lake Apopka were exposed to the estrogenic pollutants dicofol, DDT, DDE
and chloro-DDT, when a chemical plant had an extensive spill in 1980. Ten years later, researchers trying
to find out why the alligator population was dropping in the lake found higher than normal mortality
among eggs and newborn alligators. Researchers from the University of Florida in Gainesville (US),
surveying the hatching rates of alligators from various lakes, found that between 80% and 95% of
alligator eggs from Lake Apopka failed to hatch, compared to 20% to 30% for other lakes. They also
found that adolescent females had severe ovarian abnormalities and had blood estrogen levels two
times higher than normal. The male juvenile alligators were feminized, that is, they had penises one-half
to one-third the normal size, had abnormal testes and higher estrogen levels and lower testosterone
level in their blood than normal males of the same age. The researchers concluded that chemicals from
the spill not only killed developing eggs outright, but also altered the embryo’s endocrine system, which
severely limited the alligators’ ability to reproduce. The lake contained up to 15% DDT or DDE, the even
more toxic product that forms as DDT breaks down.

Studies of mammals, reptiles, birds and fish, as well as laboratory studies using rodents, primates and
cultured cells have linked exposure of a developing embryo to environmental contaminants with many
permanent health effects in the adult. The effects include abnormal blood hormone levels, reduced
fertility, altered sexual behavior, modified immune system, masculinization of female, feminization of
males (reduced testes and penis size), undescended testicles, cancers of the female and male
reproductive tract, malformed fallopian tubes, uterus and cervix, and altered bone density and
structure.

Some wildlife biologists believe that endocrine-disruption chemicals are placing many wildlife
populations at risk. Exposure to environmental contaminants has been linked to documented problems
in wildlife that include thyroid dysfunction in birds and fish; decreased fertility in birds, fish, shellfish,
and mammals; decreased hatching and gross birth deformities in birds, fish and turtles; male fish, birds
and mammals that are feminized; female fish, birds and mammals that are masculinized; and
compromised immune systems in birds and mammals. Many of these effects are manifested in species
living in or near the Great Lakes (USA), which contain many synthetic chemicals, including byproducts of
chemical production such as PCBs and pesticides. PCBs were used for a variety of purposes, including
electrical transformation and capacitors. Although their use is now severely restricted, PCBs are still
found in other electrical stations and other machinery.
The 209 different PCBs differ greatly in their estrogenic potency. Most PCBs do not readily degrade, and
they accumulate in marine animals, birds and mammals. Pesticides like DDT, DDE, dieldrin, dicofol,
kepone, mirex, etc., have been shown in tissue and biological assay to be weakly-to-moderately
estrogenic. Over the past few years since the problem became evident, a number of different assay
techniques have been developed. For egg-laying vertebrates, a technique was developed to test for the
production of vitelligenin by liver cells in response to estrogen or estrogenic substance. For mammals, a
different technique had to be developed, since they do not produce vitelligenin. A classic technique to
test for estrogenicity is the proliferation and cornification of mouse vaginal epithelium. Another
technique for testing a substance for estrogen is the induction of lactoferrin in the mouse uterus.
Lactoferrin was found to be a sensitive marker for estrogen in the mouse uterus. Soto et al. believe they
have come up with an easier and more inexpensive way of testing estrogenicity. They introduced a
proliferative assay technique (E-SCREEN) that uses human breast cancer estrogen-receptive MCF-7 cells.
These cells need estrogen to proliferate into tumors. Their assay technique produced 12 different
insecticides that tested positive for estrogen activity, the most notable being DDT, dieldrin,
methoxychlor and kepone. If the chemicals tested test positive in this assay, then they are likely to cause
proliferation in estrogen-sensitive cells in an animal’s body (i.e., endometrium, breast tissue), and this
proliferation could lead to cancer formation. Fry and Toone found that when they injected fertilized sea
bird eggs with DDT and its metabolites, the male chicks produced had varying degrees of intersexuality
depending on dosage. The female chicks had partial-to-fully developed right oviducts, instead of just left
function structure.

The discovery of a true hermaphroditic beluga whale is believed to be a result of the high
organochlorine pollution in the St. Lawrence River. Because the effects of exposure are often delayed,
they may not be fully expressed until the offspring of the exposed adult reaches maturity or even middle
age. Thus, the reproductive future for some species may be grim even if adults continue to breed and
juveniles seem healthy.

Cryptorchidism of male Florida panthers is now suspected to be caused by the eating of high pesticide-
laden prey. Cryptorchidism reduces the fertility of the animal. Additional evidence by Geliert showed
that when neonate rats were injected daily until 6 months of age with the pesticide kepone, this caused
reproductive effects on the neonate animal. The effects of the pesticide were persistent vaginal estrus,
anovulation and precocious vaginal opening. A study by Nelson showed that dieldrin and mirex
stimulates swimming in sea urchin, and lindane inhibits sperm motility. A different study showed that
mirex caused testicular cancer in humans.

Fatemeh Maleky and Somayeh Sarafpur (2020) Environmental Estrogens and their Wildlife and Human
Health Effects: A Review. Annals of Saudi Medicine. https://doi.org/10.5144/0256-4947.2001.53
https://www.annsaudimed.net/doi/full/10.5144/0256-4947.2001.53