It was 18 years ago, in 1978, when a small group of biologists studying the Elevenmile Creek in Escandia County, Florida, were "startled" (their word) to find a population of tiny mosquitofish that all appeared to be males, even though some were pregnant and were bearing normal offspring. [1] Somehow the female mosquitofish had been changed to look like males; they had been "masculinized."

Mosquitofish are a small, minnow-like species, rarely exceeding 2 inches (46 mm) in length. In normal mosquitofish, the difference between males and females is obvious. For one thing, males are about half the size of females. For another, males have a long specialized rear fin (called a gonopodium) that females lack, which the males use to insert semen into females. A few days after a female becomes fertilized, a dark spot appears on each side of her abdomen and the spot grows as pregnancy proceeds. Thus under normal circumstances, it is easy to tell male and female mosquitofish apart because they look nothing like each other. But in the Elevenmile River in Florida in 1978, the mosquitofish all had a male's gonopodium --even the pregnant females with the telltale spot. Furthermore, the female mosquitofish with a gonopodium behaved like males. They tried to copulate with other females. Further study revealed that a few members of the masculinized population were fully hermaphroditic --having sex organs of both male and female in a single individual. In 1981, a population of mosquitofish on the Fenholloway River in Florida was discovered with the same odd sexual characteristics. [2]

Odd indeed. During the past 100 years, ichthyologists (scientists who study fish) have collected and examined thousands of specimens of mosquitofish and no report has appeared of masculinization, hermaphroditism, or sex reversal (males turning into females or the other way around). [3] Yet here in two rivers in the Florida panhandle were entire populations of mosquitofish with this extraordinary physical change.

The cause was easy to find. Both rivers received enormous, dark-colored discharges from paper mills, discharges about equal to the entire flow of the rivers (25 million gallons per day (MGD) in the Elevenmile, and 50 MGD in the Fenholloway). Examining fish above and below these discharge points revealed immediately that fish above were all normal and fish below were all masculinized. An open and shut case of cause and effect.

But what in the paper mill waste was causing the problems? Paper mill waste contains between 250 and 300 chemicals, including dissolved organics, methanol, turpenes, acetone, fatty acids, cellulose decomposition products, lignins and tannins, sulphides, mercaptans, resin-acids, soaps, chlorine, and caustic soda, among others. [4]

Looking into the problem, researchers found studies dating from 1940 and 1941 in which mosquitofish had been masculinized by exposure to testosterone, the most common male sex hormone. So they reasoned that something in the paper mill waste was probably acting like a male sex hormone. But what? It seems like a simple question, but it is not.

This problem has been a scientific political football for 15 years. Two parallel tracks of research have been proceeding: dioxin and chlorinated chemicals have been investigated, and naturally-occurring hormone-mimicking chemicals from plants (phytoestrogens) have been investigated.

Dioxin Research

By 1970, there were many reports circulating which linked dioxin to various human reproductive problems, chiefly birth defects. [5] In 1980, U.S. Environmental Protection Agency [EPA] discovered dioxins and furans in paper mill waste. In 1983, dioxin was found in fish living downstream from paper mills. [6]

This was not a welcome finding. The volume of paper mill wastes is huge: in the 1980s, each ton of pulp produced about 25,000 gallons of contaminated wastewater from the chlorine-bleaching process. [7] With 73 million tons of paper products produced in 1986, [8] bleaching wastes totaled approximately 1.83 trillion gallons of chlorinated gunk per year --a total waste flow of 219 cubic meters per second [cms]. This flow is larger than some major rivers --larger than the Merrimack in Massachusetts (flow: 205 cms), larger than the James in Virginia (flow: 184 cms), larger than Brazos in Texas (flow: 207 cms), and larger than the Rogue in Oregon (flow: 182 cms). [9]

In addition to being immensely wasteful, the paper industry is politically very powerful, with 674,000 people employed in the industry (a 1986 figure), with U.S. mills producing products valued at $3.9 billion each year. [10] This powerful force has spent untold millions diverting attention away from its dioxin-laden discharges.

During the early Reagan years, top EPA officials did their part to prevent recognition of the nation's dioxin problem; EPA chief Anne Burford and her assistant, Rita Lavelle, were eventually fired for dioxin-related shenanigans, and Burford's successor, John Hernandez, resigned in disgrace after a Congress investigated his role in the altering of a report on dioxin in the Great Lakes. [11] Congress then appropriated $4 million for a nationwide study of dioxin in the environment. Preliminary data from that study, available in 1985, clearly implicated pulp and paper mills in dioxin contamination of waterways in Minnesota and Maine, and of fish living downstream from paper mills.

It was paper industry officials who muscled EPA chief William Reilly in early 1991, persuading him to initiate a multi-year "scientific reassessment" of dioxin, which EPA promptly got underway in April, 1991, and which the agency has still not concluded. During the course of that "reassessment," information has come to light showing beyond any doubt that dioxin mimics hormones and disrupts the endocrine (reproductive) system of fish, birds, mammals, and most likely humans as well. (See REHW #290, #390, #391, #414, AND #457.)

Research on Natural Phytoestrogens

The other line of research proceeded in a less politically-charged atmosphere. Scientists had first reported naturally-occurring hormone-mimicking chemicals in plants (phytoestrogens) in 1948 when female sheep became sterile after grazing on clover pastures for prolonged periods in Australia. [12]

In 1976, researchers in California had shown that certain desert plants in dry years produce chemicals that inhibit reproduction in the California quail. These researchers developed the hypothesis that plants produce such chemicals as a defense against predators; if predators eat too much of such substances, they become sterile, thus reducing the predator population. [13]

In 1979, it was shown that a common estrogen-mimicking chemical in woody plants --beta-sitosterol --could affect the reproductive system of rabbits. [14] By the mid-1980s, researchers demonstrated similar effects of beta-sitosterol on mice and rats.

In 1991, the Wingspread Conference occurred (see REHW #263 and #264), and studies of paper mill effluent began to multiply, many of them by Canadian researcher Glen Van Der Kraak, who attended Wingspread. [15,16,17,18]

Van Der Kraak and his colleagues have shown that some wild fish living downstream from pulp and paper mills reach sexual maturity much later than normal. Other species such as the lake whitefish can be sterilized by living below a pulp mill. Others have exceptionally small reproductive organs and abnormally low levels of sex hormones in the blood.

In 1995, Van Der Kraak pointed out that pulp mill waste contains 280 to 1200 parts per billion (ppb) of beta-sitosterol, from the bark of the trees being turned into pulp. It is the same beta-sitosterol that has been shown to affect the reproductive systems of rabbits, mice, rats, and sheep. Furthermore, in 1995 Van Der Kraak injected goldfish with beta-sitosterol and showed that the levels of sex hormones in their blood were significantly altered. [19] Industry is already arguing [20] that synthetic (human-created) chemicals couldn't possibly be affecting sexual reproduction and development in wildlife and humans, as many scientists now believe they are. (For example, see REHW #446, #447, #448.) No doubt, Van Der Kraak's 1995 goldfish study, emphasizing the role of naturally-occurring compounds, will become ammunition for those who need a rationale for continuing to poison wildlife and people with synthetic chlorine compounds.

However, even Van Der Kraak doesn't believe naturally-occurring phytoestrogens explain all the problems seen in wildlife exposed to paper mill wastes. "I don't think beta-sitosterol is going to explain the entire picture," Van Der Kraak told Janet Raloff of SCIENCE NEWS. [21] Fish downstream from pulp mills frequently exhibit detoxifying liver enzymes, "and we're not seeing that with beta-sitosterol," he says. "So there must be other chemicals the fish are exposed to--such as some chlorinated organics--[activating] those enzymes," he says. In other words, the trail once again leads back to dioxin and other chlorine-containing wastes.

                                                                         --Peter Montague
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[1] William P. Davis and Stephen A. Bortone, "Effects of Kraft Mill Effluent on the Sexuality of Fishes: An Environmental Early Warning?" in Theo Colborn and Coralie Clement, editors, CHEMICALLY-INDUCED ALTERATIONS IN SEXUAL AND FUNCTIONAL DEVELOPMENT: THE WILDLIFE/HUMAN CONNECTION [Advances in Modern Environmental Toxicology Vol. XXI] (Princeton, N.J.: Princeton Scientific Publishing Co., 1992), pgs. 113-127.

[2] W. Mike Howell and others, "Abnormal Expression of Secondary Sex Characters in a Population of Mosquitofish, GAMBUSIA AFFINIS HOLBROOKI: Evidence for Environmentally-induced Masculinization," COPEIA Vol. 4 (1980), pgs. 676-681.

[3] W. Mike Howell and Thomas E. Denton, "Gonopodial morphogenesis in female mosquitofish, GAMBUSIA AFFINIS AFFINIS, masculinized by exposure to degradation products from plant sterols," ENVIRONMENTAL BIOLOGY OF FISHES Vol. 24, No. 1 (1989), pgs. 43-51.

[4] Leena R. Suntio and others, "A Review of the Nature and Properties of Chemicals Present in Pulp Mill Effluents," CHEMOSPHERE Vol. 17, No. 7 (1988), pgs. 1249-1290.

[5] See, for example, Nicholas Wade, "Viets and vets fear herbicide health effects," SCIENCE Vol. 204 (May 25, 1970), pg. 817. Carol Van Strum, A BITTER FOG (San Francisco: Sierra Club Books, 1983), in footnotes 6, 9, 10, and 11 (found on pgs. 258 and 259) of chapter 2 lists other reports of dioxin problems known in 1970.

[6] Carol van Strum and Paul Merrell, NO MARGIN OF SAFETY: A PRELIMINARY REPORT ON DIOXIN POLLUTION AND THE NEED FOR EMERGENCY ACTION IN THE PULP AND PAPER INDUSTRY (Washington, D.C.: Greenpeace USA, 1987). See footnotes 14 and 15 on pg. V-7.

[7] John Andelin and others, TECHNOLOGIES FOR REDUCING DIOXIN IN THE MANUFACTURE OF BLEACHED WOOD PULP; BACKGROUND PAPER [OTA-BP-O-54] (Washington, D.C.: U.S. Congress, Office of Technology Assessment, May 1989). See pg. 55.

[8]] See John Andelin and others, cited above, pg. 5.

[9] River flows are from Frits van der Leeden and others, editors, THE WATER ENCYCLOPEDIA [SECOND EDITION] (Chelsea, Michigan: Lewis Publishers, 1990), pgs. 174-175.

[10] See John Andelin and others, cited above, pg. 5.

[11] See Carol van Strum and Paul Merrell, cited above, Chapter V.

[12] Rami S. Kaldas and Claude L. Hughes, Jr., "Reproductive and General Metabolic Effects of Phytoestrogens in Mammals," REPRODUCTIVE TOXICOLOGY Vol. 3 (1989), pgs. 81-89.

[13] A. Starker Leopold and others, "Phytoestrogens: Adverse Effects on Reproduction in California Quail," SCIENCE Vol. 191 (January 9, 1976), pgs. 98-100.

[14] S.A. Ghannudi and others, "Adverse Effects of Phytoestrogens III. The Effect of Beta-sitosterol on the Ovarian Structures of Immature Rabbits," THE LIBYAN JOURNAL OF SCIENCE Vol. 9a (1979), pgs. 1-12.

[15] K.R. Munkittrick and others, "Reproductive Dysfunction and MFO Activity in Three Species of Fish Exposed to Bleached Kraft Mill Effluent at Jackfish Bay, Lake Superior," WATER POLLUTION RESEARCH JOURNAL OF CANADA Vol. 27, No. 3 (1992), pgs. 439-446.

[16] M.E. McMaster and others, "Changes in hepatic mixed-function oxygenase (MFO) activity, plasma steroid levels and age at maturity of a white sucker (CATOSTOMUS COMMERSONI) population exposed to bleached kraft pulp mill effluent," AQUATIC TOXICOLOGY Vol. 21 (1991), pgs. 199-218.

[17] G.J. Van Der Kraak and others, "Exposure to Bleached Kraft Pulp Mill Effluent Disrupts the Pituitary-Gonadal Axis of White Sucker at Multiple Sites," TOXICOLOGY AND APPLIED PHARMACOLOGY Vol. 115 (1992), pgs. 224-233.

[18] K.R. Munkittrick and others, "Changes in Maturity, Plasma Sex Steroid Levels, Hepatic Mixed-Function Oxygenase Activity, and the Presence of External Lesions in Lake Whitefish (COREGONUS CLUPEAFORMIS) Exposed to Bleached Kraft Mill Effluent," CANADIAN JOURNAL OF FISHERIES AND AQUATIC SCIENCES Vol. 49 (1992), pgs. 1560-1569.

[19] Deborah L. MacLatchy and Glen J. Van Der Kraak, "The Phytoestrogen Beta-Sitosterol Alters the Reproductive Endocrine Status of Goldfish," TOXICOLOGY AND APPLIED PHARMACOLOGY Vol. 134 (1995), pgs. 305-312.

[20] Stephen H. Safe, "Environmental and Dietary Estrogens and Human Health: Is There a Problem?" ENVIRONMENTAL HEALTH PERSPECTIVES Vol. 103, No. 4 (April, 1995), pgs. 346-351.

[21] Janet Raloff, "How paper mill wastes may imperil fish," SCIENCE NEWS Vol. 148 (November 4, 1995), pg. 295.

Descriptor terms: paper; paper industry; chlorine; dioxin; phytoestrogens; estrogen; reproductive disorders; birth defects; water pollution; fl; fish; wildlife; endocrine disrupters; hormones; anne burford; rita lavelle; john hernandez; ronald reagan; epa; dioxin reassessment; beta-sitosterol; glen van der kraak;