Effects of environmental estrogens on animals in Israel: implications for effects on humans.

In the last decade interest has increased in the possible effects of endocrine disruptor compounds. Numerous papers have appeared as to their possible effects on humans, but definitive effects are hard to demonstrate. In the field of animal husbandry, however, the effects of the endocrine disruptors are well documented. This paper discussed the effects of the endocrine disruptors seen in animals and the possible implications for humans. The areas considered are reproductive disorders, premature udder/breast development, prolapsed oviduct/uterus, scrotal atrophy, and skewed sex ratio.

The environmental compartments of environmental hormones.

Compounds with estrogenic activity are ubiquitous in nature. Phytoestrogens and steroidal estrogens are found naturally in the food, in particular legumes and milk products. Naturally occurring steroidal estrogens and synthetic estrogens such as ethinylestradiol are constantly excreted into the environment as the result of the release of animal and human waste. As the result of this constant exposure, testosterone, estrogens, and ethinylestradiol are readily detectable in every stream examined in Israel. The concentrations observed of the estrogenic compounds are physiological, i.e., at these concentrations fish reproductive function can be affected. Estrogenic compounds do not usually reach the groundwater but testosterone percolates through the soil and is frequently found in the groundwater. In the dry season, there are no natural or synthetic steroids in the streams of the Jordan River watershed. However, at the beginning of the rainy season, both estrogen and testosterone reach the surface waters in the runoff from fields with manure, whereas ethinylestradiol reaches the streams in overflows from oxygenation ponds used to treat sewage prior to irrigation.

Behavioral changes, stress, and survival following reintroduction of Persian fallow deer from two breeding facilities.

Reintroductions often rely on captive-raised, naïve animals that have not been exposed to the various threats present in natural environments. Wild animals entering new areas are timid and invest much time and effort in antipredator behavior. On the other hand, captive animals reared in predator-free conditions and in close proximity to humans may initially lack this tendency, but can reacquire some antipredator behavior over time. We monitored the changes in antipredator-related behaviors of 16 radio-collared Persian fallow deer (Dama mesopotamica) reintroduced to the Soreq Valley in Israel from 2 breeding facilities: one heavily visited by the public (The Biblical Zoo of Jerusalem, Israel) and the other with reduced human presence (Hai-Bar Carmel, Israel). We monitored each individual for up to 200 days after release, focusing on flush and flight distance, flight mode (running or walking), and use of cover. In addition, we compared fecal corticosterone (a stress-related hormone) from samples collected from known animals in the wild to samples collected in the breeding facilities. Reintroduced individuals from both origins exhibit increased flush distance over time; flush and flight distances were larger in individuals from Hai-Bar; use of cover increased with time, but was greater in Hai-Bar Carmel animals; corticosterone levels were significantly higher in fecal samples from reintroduced animals than in samples from captive animals; and Hai-Bar Carmel animals had an 80% survival rate over the 200 days, whereas no animals from the Biblical Zoo of Jerusalem survived. Reintroduced Persian fallow deer reacquired antipredator behaviors after the release, but the process was slow (months) and differences between conditions at the breeding facilities that were seemingly benign (e.g., number of visitors and other human related activities) influenced this process and consequently affected the success of the reintroduction. Captive breeding facilities for the purpose of reintroduction should minimize anthropogenic disturbances.

Washout of accumulated testosterone in a watershed.

Testosterone is constantly excreted into the environment by both human and animal sources but little is known about how it is transported in the environment. In this study, testosterone was measured in 15 sites in the Upper Jordan Valley after major rain events (238 samplings) for two consecutive rain seasons. The area consists of small farms, cattle pasture, fish ponds with some urban development. One liter samples were extracted on solid phase columns and the eluates measured using specific radio-immunoassay for testosterone and estrogen (estradiol + estrone). The first rain season was the first above average season after a 3-year period of well below average rainfall. It was found in the rain season of 2001/2002, that following a rain sequence of 131 mm/week there was an initial large increase in the concentration of testosterone (maximum 6 ng/l) accompanied by high estrogen (maximum 6 ng/l), which then gradually declined to non-detectable levels (< 0.3 ng/l) over a period of 3 months. These peaks originated from runoff from cattle pasture and fish pond effluent. Later peaks consisted only of testosterone that was moderately associated with sulfate (r2 = 0.53, P < 0.05) and somewhat associated with total phosphorus (r2 = 0.49, P < 0.1) indicating that the origin was leaching from the sulfurous peat soil. In the following rainy season, which had recorded rainfalls, no testosterone peaks above 1 ng/l were seen. We conclude that the testosterone accumulated in the Upper Jordan Valley was washed out in two stages, first as surface runoff from cattle pasture and then as discharge from the soil.

Effect of reproductive status on intraocular pressure in cats.

OBJECTIVE: To measure intraocular pressure (IOP) and progesterone concentrations in cats and to examine their reproductive organs to determine whether reproductive status affects IOP in cats. ANIMALS: 75 sexually intact domestic shorthair cats scheduled to be neutered, including 28 males, 21 females not in estrus, 13 females in estrus, and 13 pregnant females. PROCEDURES: Applanation tonometry was conducted to measure IOP and radioimmunoassay was used to determine progesterone concentrations. Reproductive organs were examined at time of surgery. RESULTS: The IOP in female cats that were in estrus was significantly higher than IOP in female cats that were not in estrus. Progesterone concentrations significantly affected IOP in pregnant cats. CONCLUSIONS AND CLINICAL RELEVANCE: In cats, IOP is affected by changes in reproductive status. Such changes should be considered when interpreting tonometry results in this species.

Reproductive characteristics and steroid levels in the scleractinian coral Oculina patagonica inhabiting contaminated sites along the Israeli Mediterranean coast.

In this study we compared reproductive characteristics and steroid hormone levels in the non-indigenous scleractinian coral, Oculina patagonica, inhabiting contaminated vs. uncontaminated reference sites along the Israeli Mediterranean coast. Our results indicate significantly higher steroid levels in both seawater and coral tissue samples from contaminated sites as compared to reference sites, suggesting that corals tend to accumulate steroids from the surrounding waters. Despite their higher steroid levels, corals from the contaminated sites showed reproductive potential comparable to those of the reference sites. Interestingly, a clear distinction could be seen between corals exposed to pollution for long vs. short durations, with the latter showing a failure to complete gametogenesis. This suggests that reproduction in O. patagonica is susceptible to acute rather than chronic stress. The involvement of adjustment/adaptation processes may explain this species tolerance, and may reflect the ability of O. patagonica to successfully invade new areas in the Mediterranean Sea.

Use of sperm plasmid DNA lipofection combined with REMI (restriction enzyme-mediated insertion) for production of transgenic chickens expressing eGFP (enhanced green fluorescent protein) or human follicle-stimulating hormone.

Linearized p-eGFP (plasmid-enhanced green fluorescent protein) or p-hFSH (plasmid human FSH) sequences with the corresponding restriction enzyme were lipofected into sperm genomic DNA. Sperm transfected with p-eGFP were used for artificial insemination in hens, and in 17 out of 19 of the resultant chicks, the exogenous DNA was detected in their lymphocytes as determined by PCR and expressed in tissues as determined by (a) PCR, (b) specific emission of green fluorescence by the eGFP, and (c) Southern blot analysis. A complete homology was found between the Aequorea Victoria eGFP DNA and a 313-bp PCR product of extracted DNA from chick blood cells. Following insemination with sperm lipofected with p-hFSH, transgenic offspring were obtained for two generations as determined by detection of the transgene for human FSH (PCR) and expression of the gene (RT-PCR and quantitative real-time PCR) and the presence of the protein in blood (radioimmunoassay). Data demonstrate that lipofection of plasmid DNA with restriction enzyme is a highly efficient method for the production of transfected sperm to produce transgenic offspring by direct artificial insemination.

Transport of testosterone and estrogen from dairy-farm waste lagoons to groundwater.

Although concentrated animal feeding operations constantly generate physiologically active steroidal hormones, little is known of their environmental fate. Estrogen and testosterone concentrations in groundwater and their distribution in sediments below a dairy-farm wastewater lagoon were therefore determined and compared to a reference site located upgradient of the farm. Forward simulations of flow as well as estrogen and testosterone transport were conducted based on data from the sediment profile obtained during drilling of a monitoring well belowthe dairy-farm waste lagoon. Testosterone and estrogen were detected in sediments to depths of 45 and 32 m, respectively. Groundwater samples were directly impacted by the dairy farm, as evidenced by elevated concentrations of nitrate, chloride, testosterone, and estrogen as compared to the reference site. Modeling potential transport of hormones in the vadose zone via advection, dispersion, and sorption could not explain the depths at which estrogen and testosterone were found, suggesting that other transport mechanisms influence hormone transport under field conditions. These mechanisms may involve interactions between hormones and manure as well as preferential flow paths, leading to enhanced transport rates. These types of interactions should be further investigated to understand the processes regulating hormone transport in the subsurface environment and parametrized to forecast long-term fate and transport of steroidal hormones.

Monitoring of natural and synthetic hormones in a polluted river.

Natural (estradiol, estrone, testosterone, estriol) and synthetic hormones (ethinylestradiol) are constantly excreted into the environment from human and animal sources but little is known of their transport. The purpose of this study was to determine how far along a 100 km river course that hormones could be detected after contamination with sewage effluent or fishpond effluent. Fourteen sites in the Lower Jordan River drainage were sampled (two sites above the sewage effluent contamination, eight sites below the contamination and four tributaries) before and after the dry season of 2002 (Spring and Fall). Samples were tested for testosterone, estrogen (estrone and estradiol combined), estriol, ethinylestradiol, ammonia and fecal coli. It was found that the fecal coli count dropped exponentially (from 250,000 to 60/100 ml3) and the ammonia dropped from 15 to less than 1mg/l over the initial 25 km stretch. Over the same stretch, the hormone values declined by half from their maximum values for testosterone (3.3 ng/l), estriol (8.8 ng/l), ethinylestradiol (6.1 ng/l), and estrogen (4.9 ng/l). From 67 to 100 km mark, testosterone (4.8 ng/l) and estrogen (2.4 ng/l) were still elevated while ethinylestradiol and estriol were >or=1.5 ng/l. The high level of testosterone and estrogen between 67 and 100 km marks was probably due to major discharge from fishponds between 23 and 27 km marks. Levels of ethinylestradiol above 1 ng/l, a level which can affect fish, was seen in 70% (12/16) of the samples tested. The data suggest that hormones in readily measured quantities can be transported considerable distances from the source of pollution.