Comparison of three enzyme immunoassays for measuring 17beta-estradiol in flushed dairy manure wastewater

Natural steroidal estrogens are an environmental concern because low nanogram per liter concentrations in water can adversely affect aquatic vertebrate species by disrupting the normal function of their endocrine systems. There is a critical need to accurately measure estrogens in dairy wastes, a potential source of estrogens such as 17beta-estradiol, to assess the risk of estrogen contamination of agricultural drainage waters resulting from land application. Commercially available enzyme immunoassay (EIA) kits have been used for measuring 17beta-estradiol in livestock manure, but it is not known if different EIAs provide similar results. We compared three EIAs by measuring 17beta-estradiol in two samples of flushed dairy manure wastewater (FDMW). The measured concentrations of 17beta-estradiol in FDMW differed according to the immunoassay used. The differences were attributed to a matrix interference associated with coextracted humic substances. Future research should develop methods that enable routine measurement of 17beta-estradiol in livestock wastes by more conclusive analytical techniques such as gas chromatography-mass spectrometry.     

Sorption, mobility, and transformation of estrogenic hormones in natural soil

Potent estrogenic hormones are consistently detected in the environment at low concentration, yet these chemicals are strongly sorbed to soil and are labile. The objective of this research was to improve the understanding of the processes of sorption, mobility, and transformation for estrogens in natural soils, and their interaction. Equilibrium and kinetic batch sorption experiments, and a long-term column study were used to study the fate and transport of 17beta-estradiol and its primary metabolite, estrone, in natural soil. Kinetic and equilibrium batch experiments were done using radiolabeled 17beta-estradiol and estrone. At the concentrations used, it appeared that equilibrium sorption for both estrogens was achieved between 5 and 24 h, and that the equilibrium sorption isotherms were linear. The log K(oc) values for 17beta-estradiol (2.94) and estrone (2.99) were consistent with previously reported values. Additionally, it was found that there was rate-limited sorption for both 17beta-estradiol (0.178 h(-1)) and estrone (0.210 h(-1)). An approximately 42 h long, steady-flow, saturated column experiment was used to study the transport of radiolabeled 17beta-estradiol, which was applied in a 5.00 mg L(-1) solution pulse for 44 pore volumes. 17beta-estradiol and estrone were the predominant compounds detected in the effluent. The effluent breakthrough curves were asymmetric and the transport modeling indicated that sorption was rate-limited. Sorption rates and distributions of the estrogens were in agreement between column and batch experiments. This research can provide a better link between the laboratory results and observations in the natural environment.     

Free and conjugated estrogen exports in surface-runoff from poultry litter-amended soil

Land application of animal manures such as poultry litter is a common practice, especially in states with surplus manure. Past studies have shown that animal manure may contain estrogens, which are classified as endocrine-disrupting chemicals and may pose a threat to aquatic and wildlife species. We evaluated the concentrations of estrogens in surface runoff from experimental plots (5 x 12 m each) receiving raw and pelletized poultry litter. We evaluated the free (estrone, E1; 17beta-estradiol, E2beta; estriol, E3) and conjugate forms (glucuronides and sulfates) of estrogens, which differ in their toxicity. Sampling was performed for 10 natural storm events over a 4-mo period (April-July 2008). Estrogen concentrations were screened using enzyme-linked immunosorbent assay (ELISA), followed by quantification using liquid chromatography with tandem mass spectrometry (LC/MS/MS). Concentrations of estrogens from ELISA were much higher than the LC/MS/MS values, indicating crossreactivity with organic compounds. Exports of estrogens were much lower from soils amended with pelletized poultry litter than the raw form of the litter. No-tillage management practice also resulted in a lower export of estrogens with surface runoff compared with reduced tillage. The concentrations and exports of conjugate forms of estrogens were much higher than the free forms for some treatments, indicating that the conjugate forms should be considered for a comprehensive assessment of the threat posed by estrogens.     

Persistence of testosterone and 17beta-estradiol in soils receiving swine manure or municipal biosolids

Natural and synthetic steroidal hormones can be carried to agricultural soil through fertilization with municipal biosolids, livestock manure, or poultry manure. The persistence and pathways of dissipation of [4-(14)C]-testosterone and of [4-(14)C]-17beta-estradiol in organic-amended soils were investigated using laboratory microcosms. Testosterone dissipation was investigated over a range of amendment concentrations, temperatures, and soil types. Under all conditions the parent compound and transformation products were dissipated within a few days. Addition of swine manure slurry to soil hastened the transformation of testosterone and 17beta-estradiol to the corresponding less hormonally active ketones, 4-androstene-3,17-dione and estrone. Two other testosterone transformation products, 5alpha-androstan-3,17-dione and 1,4-androstadiene-3,17-dione, were also detected. Experiments with sterilized soil and sterilized swine manure slurry suggested that the transformation of (14)C-labeled hormonal parent compounds was mainly caused by microorganisms in manure slurry, while mineralization of the hormones to (14)CO(2) required viable soil microorganisms. Organic amendments transiently inhibited the mineralization of [4-(14)C]-testosterone, perhaps by inhibiting soil microorganisms, or by enhancing sorption and reducing the bioavailability of testosterone or transformation products. Overall, organic amendments influenced the pathways and kinetics of testosterone and estradiol dissipation, but did not increase their persistence.     

Mineralization of hormones in breeder and broiler litters at different water potentials and temperatures

When poultry litter is landspread, steroidal hormones present in the litter may reach surface waters, where they may have undesirable biological effects. In a laboratory study, we determined the mineralization of [4-14C]-labeled 17beta-estradiol, estrone, and testosterone in breeder litter at three different water potentials (-56, -24, and -12 MPa) and temperatures (25, 35, and 45 degrees C), and in broiler litter at two different water potentials (-24 and -12 MPa) and temperatures (25 and 35 degrees C). Mineralization was similar in both litters and generally increased with increasing water content and decreasing temperature. After 23 wk at -24 MPa, an average of 27, 11, and <2% of the radiolabeled testosterone applied to breeder litter was mineralized to 14CO2 at 25, 35, and 45 degrees C, respectively. In contrast, mineralization of the radiolabeled estradiol and estrone was <2% after 25 wk at all water potentials, except after 17 wk at 25 degrees C and -12 MPa, where up to 5.9% of the estradiol and 7.8% of the estrone was mineralized. The minimal mineralization suggests that the litters may still be potential sources of hormones to surface and subsurface waters.     

Estrogenic Compounds Downstream From Three Small Cities in Eastern Nebraska: Occurrence and Biological Effect1

Abstract: Recent studies have detected estrogenic compounds in surface waters in North America and Europe. Furthermore, the presence of estrogenic compounds in surface waters has been attributed, in some cases, to the discharge of wastewater treatment plant (WWTP) effluent. The primary objective of the current study was to determine if WWTP effluent contributes estrogens to the surface waters of Nebraska. A second objective of this study was to determine if estrogens were found in concentrations sufficient enough to manifest feminizing effects on fish. These objectives were satisfied by deploying polar organic chemical integrative samplers (POCIS) and caged fathead minnows at eight field sites. Deployment sites included: three reference sites (Pawnee Creek, the Little Blue River, and the Middle Loup River), two sites upstream of the WWTPs at Grand Island and Columbus, and three sites downstream of the WWTPs at Grand Island, Columbus, and Hastings. Following the seven day deployments, POCIS extracts were analyzed for estrone, 17β‐estradiol, estriol and 17α‐ethinylestradiol using liquid chromatography tandem mass spectrometry (LC/MS/MS). 17β‐estradiol was detected in POCIS from six of the eight field sites with the greatest quantities recovered in POCIS deployed downstream from the Grand Island and Hastings WWTPs. Estrone was detected only in the POCIS deployed downstream from the Grand Island and Hastings WWTPs. Estrogenic effects were detected in caged minnows analyzed for the hepatic mRNA expression of two estrogen‐responsive genes, vitellogenin (vg1) and estrogen receptor α (ERα). Fish deployed at the site where the greatest quantities of estrogens were recovered (Hastings) had significantly higher expression of both vg1 and ERα than fish deployed at any of the other sites. These results confirm that WWTP effluent contributes biologically significant levels of estrogens to Nebraska surface waters.     

Decrease in water-soluble 17beta-Estradiol and testosterone in composted poultry manure with time

Little attention has been paid to the environmental fate of the hormones 17beta-estradiol and testosterone excreted in animal waste. Land application of manure has a considerable potential to affect the environment with these endocrine disrupting compounds (EDCs). Composting is known to decompose organic matter to a stable, humus-like material. The goal of the present study was to quantitatively assess levels of water-soluble 17beta-estradiol and testosterone in composting chicken manure with time. Chicken layer manure was mixed with hay, straw, decomposed leaves, and starter compost, adjusted to approximately 60% moisture, and placed into a windrow. A clay-amended windrow was also prepared. Windrows were turned weekly, and temperature, oxygen, and CO(2) in the composting mass were monitored for either 133 or 139 d. Commercial enzyme immunoassay kits were used to quantitate the levels of 17beta-estradiol and testosterone in aqueous sample extracts. Water-soluble quantities of both hormones diminished during composting. The decrease in 17beta-estradiol followed first-order kinetics, with a rate constant k = -0.010/d. Testosterone levels declined at a slightly higher rate than 17beta-estradiol (i.e., k = -0.015/d). Both hormones could still be measured in aqueous extracts of compost sampled at the conclusion of composting. The decline in water-soluble 17beta-estradiol and testosterone in extracts of clay-amended compost was not statistically different from normal compost. These data suggest that composting may be an environmentally friendly technology suitable for reducing, but not eliminating, the concentrations of these endocrine disrupting hormones at concentrated animal operation facilities.     

Optimizing phosphorus characterization in animal manures by solution phosphorus-31 nuclear magnetic resonance spectroscopy

A procedure involving alkaline extraction and solution 31P nuclear magnetic resonance (NMR) spectroscopy was developed and optimized for the characterization of P in animal manures (broiler, swine, beef cattle). Inclusion of ethylenediaminetetraacetic acid (EDTA) in the alkaline extraction solution recovered between 82 and 97% of the total P from the three manures, which represented a significant improvement on recovery in NaOH alone. Low concentrations of paramagnetic ions in all manure extracts meant that relatively long delay times (> 5 s) were required for quantitative analysis by solution 31P NMR spectroscopy. The manures contained inorganic orthophosphate, orthophosphate monoesters, orthophosphate diesters, and inorganic polyphosphates, but results were markedly influenced by the concentration of NaOH in the extractant, which affected both spectral resolution and the apparent P composition of the extracts. For example, extraction of swine manure and broiler litter with 0.5 M NaOH + 50 mM EDTA produced remarkable spectral resolution that allowed accurate quantification of the four signals from phytic acid, the major organic P compound in these manures. In contrast, more dilute NaOH concentrations produced considerable line broadening that obscured individual signals in the orthophosphate monoester region of the spectra. Spectral resolution of cattle manure extracts was relatively unaffected by NaOH concentration. Improvements in spectral resolution of more concentrated NaOH extracts were, however, compromised by the disappearance of phospholipids and inorganic polyphosphates, notably in swine and cattle manure extracts, which indicated either degradation or a change in solubility. The optimum extraction conditions will therefore vary depending on the manure type and the objectives of the study. Phytic acid can be accurately quantified in swine manure and broiler litter by extraction with 0.5 M NaOH + 50 mM EDTA, while a more dilute NaOH concentration should be used for complete P characterization or comparison among different manure types.     

Estrogenic activity in the environment: municipal wastewater effluent, river, ponds, and wetlands

Estrogenic activity of regional water samples was evaluated. Samples obtained from wetlands and ponds involved in various agricultural land uses, from three river sites over four seasons, and from municipal wastewater effluent held in storage lagoons were evaluated. The estrogen-responsive cell line MCF-7 BOS was used in the E-screen assay to determine 17beta-estradiol equivalents (E2 Eq) of water samples extracted by solid-phase extraction. Estrogenic activity in surrounding wetlands and ponds from different land uses was not different, with 10(-12) M E2 Eq (0.3 ppt). Estrogenic activity of Red River samples was within the same range as wetland-pond samples. The highest activity was found downstream from municipal wastewater treatment effluent discharge sites, in winter when river flow was lowest (approximately 6 x 10(-13) M E2 Eq). Results showed that 7 of 20 wetland-pond samples and 5 of 12 river samples were below the limits of quantitation (approximately 3 x 10(-14) M E2 Eq). Toxicity was found in fall and summer river samples upstream from municipal wastewater release sites. The timing of toxicity did not coincide to the presence of elevated fecal coliforms. Estrogenic activity in wastewater effluent from lagoons decreased over time (approximately 25 to 5 x 10(-13) M E2 Eq) with an apparent half-life of 8 d for one lagoon. The median concentration of detectable estrogenic activity in regional water samples was approximately 50-fold less than the median 17beta-estradiol concentration of estradiol detected in some U.S. streams in previous studies.     

Dissolved Organic Carbon and Estrogen Transport in Surface Runoff From Agricultural Land Receiving Poultry Litter1

Dutta, Sudarshan, Shreeram Inamdar, Jerry Tso, Diana S. Aga, and J. Tom Sims, 2012. Dissolved Organic Carbon and Estrogen Transport in Surface Runoff from Agricultural Land Receiving Poultry Litter. Journal of the American Water Resources Association (JAWRA) 48(3): 558-569. DOI: 10.1111/j.1752-1688.2011.00634.x Abstract: Dissolved organic carbon (DOC) provides a reactive substrate for the transport of organic contaminants with runoff. Very few studies have investigated the export of DOC from agricultural land, especially those receiving manure applications. We investigated exports of DOC in surface runoff from agricultural fields receiving various treatments of poultry litter (raw vs. pelletized). In addition, we also investigated how estrogens in runoff were associated with DOC. Different forms of estrogens studied were: estrone, 17β-estradiol, estriol, and their conjugates. Experimental agricultural plots were 12 m × 5 m long and had reduced tillage and no-till management practices. The aromatic content of DOC was characterized using specific ultraviolet absorbance (SUVA). Flow-weighted concentrations of DOC and SUVA in surface runoff from plots with poultry litter were significantly (p ≤ 0.10) greater than the control (no litter) plots. Compared to pelletized poultry litter, reduced-tillage plots with raw litter yielded higher DOC concentrations and SUVA values. No significant differences (p ≥ 0.10) in DOC and SUVA were observed between litter treatments for plots with no-till. Total estrogen concentrations (including all forms) were positively and significantly (p ≤ 0.10) correlated with DOC. These results can help select and guide agricultural management practices that can reduce the exports of DOC and associated contaminant from agricultural land receiving manure applications.     

Statewide Survey of Hormones and Antibiotics in Surface Waters of Delaware

Water‐quality surveys have confirmed the presence of hormones and antibiotics in surface waters of the United States, which may be of concern to aquatic life. We investigated the concentrations of hormones and antibiotics in surface waters of the state of Delaware to determine – how they compared against environmental thresholds, how they varied across the state, and if they were correlated with land use type. Fifty surface water locations were sampled during early spring and late summer. Water samples were initially screened with ELISA followed by analysis with LC/MS/MS. The measured ranges of hormone concentrations were: 0‐3.71 ng/l for estrone, 0‐4.65 ng/l for estrone‐3‐sulfate, and 0‐6.27 ng/l for 17β‐estradiol. The measured ranges of antibiotics were: 0‐3.30 ng/l for sulfamerazine, 0‐10.74 ng/l for sulfamethoxazole, and 0‐2.29 ng/l for tetracycline. The predicted no‐effect concentration (PNEC) for estrone was exceeded for three samples and the PNEC for 17β‐estradiol was exceeded for 11 samples. In general, concentrations and detection frequencies were lower in the summer than the spring. The highest concentrations of hormones and antibiotics were spatially distributed in agricultural and urban areas; however, the correlations between land use type and the concentrations were weak. This study was the first statewide survey of hormones and antibiotics for Delaware and provided important baseline data on these emerging contaminants.     

The Effect of Estrogens,River Water,and Treated Wastewater on Vitellogenin Induction of Japanese Medaka1

Abstract: The objective of this study is to evaluate the effect of estrogens on fish endocrine disruption in river water and treated wastewater. Endogenous estrogen estrone (E1), 17β‐estradiol (E2), river water, and treated wastewater were used for exposure tests on male Japanese medaka. Vitellogenin induction in male medaka was regarded as the endpoint of endocrine disruption. The effective concentrations of E1 and E2 on vitellogenin induction in medaka were evaluated by breeding medaka for 14 days in tanks with various concentrations of E1 or E2. Vitellogenin induction increased with elapse of time during exposure, with higher estrogen concentrations causing greater vitellogenin induction. According to the test results, the lowest observed effect concentrations (LOECs) of E1 and E2 were estimated to be 31.6 and 5.0 ng/l, respectively; and the E2 equivalent (EEQ) LOEC value of E1 was 5.2 ng‐E2/l, derived by multiplying the relative potency of E1 by that of E2 as estimated in this study. In continuous exposure tests using river water or treated wastewater, the estrogenic activity, an index of total estrogenic potential measured by yeast screen assay and expressed as EEQ, varied widely during the exposure tests, and significant vitellogenin induction was observed after several days of high levels of estrogenic activity in water. Vitellogenin concentration tended to significantly increase if EEQ exceeded the level of 5 ng‐E2/l. The threshold value was substantially the same as the results for E1 and E2 exposure tests of medaka. Consequently, EEQ over 5 ng‐E2/l was revealed to have the potential to cause endocrine disruption of male medaka. As estrogenic activity exceeding 5 ng‐E2/l was observed in some rivers in Japan, total estrogenic potential needs to be further decreased at wastewater treatment plants to prevent fish endocrine disruption.     

南方某水厂处理工艺过程中甾体雌激素的变化规律

对南方某水厂处理工艺过程中甾体雌激素进行了研究。采用固相萃取-高效液相色谱法对水源水和各处理工艺的出水进行了检测,探讨了该水厂不同处理工艺对甾体雌激素的去除效果。结果表明：各处理工艺段水样甾体雌激素均有检出,且浓度较高。工艺中甾体雌激素均有不同程度的去除,甾体雌激素去除率为19.51%～50%,研究表明,现有工艺（预氯化＋絮凝沉淀工艺）不能有效去除甾体雌激素。     

固相萃取-超高效液相色谱-串联质谱法测定地表水中5种磺胺类抗生素

建立了固相萃取-超高效液相色谱-串联质谱对水中5种磺胺类抗生素的测定分析方法。将水样经固相萃取后进样,经ACQUITY UPLC BEH C18(2.1 mm×100 mm,1.7μm)分离后,采用串联质谱电喷雾电离(ESI+),多反应监测(MRM)模式检测,内标法定量。方法的线性范围为5.0~100.0μg/L,线性相关系数均大于0.995,样品100倍浓缩后,检出限低至0.009μg/L;在5.0~100.0μg/L范围内,高中低3个水平的加标回收率为80.2%~90.7%,相对标准偏差(RSD)为4.3%~12.0%。该方法能够满足《国务院关于印发水污染防治行动计划的通知》限制使用抗生素等化学药品开展专项整治的需要,是一种高效、准确、灵敏度高的方法。     

Sorption of 17β-estradiol to pig slurry separates and soil in the soil-slurry environment

Contamination of freshwater by estrogens from manure applied to agricultural land is of grave concern because of the potentially harmful effects on aquatic life and human health. Recent developments in liquid manure (slurry) management include partial removal of particulate slurry dry matter (PSDM) by separation technologies, which may also remove parts of the estrogens and enhance infiltration of the slurry on field application and hence the interaction between estrogens and the soil matrix. This study investigated how 17β-estradiol (E2), a natural estrogen commonly found in pig manure, sorbs to agricultural soils, to different size fractions of pig slurry separates, and to soils amended with each size fraction to simulate conditions in the soil-slurry environment. A crude fiber fraction (SS1) was prepared by sieving (<500 μm) the solids removed by an on-farm separation process. Three other size fractions (SS2 > SS3 > SS4) were prepared from the liquid fraction of the separated slurry by sedimentation and centrifugation. Sorption experiments were conducted in 0.01 mol L(-1) CaCl(2) and in natural pig urine matrix. Sorption in 0.01 mol L(-1) CaCl(2) was higher than that in pig urine for all solids used. Sorption of E2 to soil increased with its organic carbon content for both liquid phases. The solid-liquid partition coefficients of slurry separates were 10 to 30 times higher than those of soils, but the organoic carbon normalized partition coefficient values, reflecting sorption per unit organic carbon, were lower for slurry separates. Mixing slurry separates with soil increased the sorption of E2 to the solid phase significantly in the order: SS1 < SS3 < SS2 for both liquid phases. In contrast, SS4 reduced the sorption of E2 to the solid phase by increasing the sorption to suspended or dissolved organic matter. The study suggested that potentially 50 to 75% of E2 in slurry can be removed from the liquid fraction of slurry by physical separation.     

Comparison of triticonazole dissipation after seed or soil treatment

This study examined the long-term fate of the fungicide triticonazole (TTZ; 5-[(4-chlorophenyl)methylene]-2,2-dimethyl-1-(1H-1,2,4-triazol-1-ylmethyl)cydopentanol) applied at a normal field dose (8.9 g ha(-1)) via seed treatment, which is the normal alternative in practice. The TTZ was applied to wheat (Triticum aestivum L.) grains as a disinfectant before sowing or spraying on bare soil for comparison and reference to the seed treatment. The seeds were germinated and grown in pots in a greenhouse at 22 +/- 3 degrees C. The dissipation of TTZ was studied by gas chromatography-mass spectrometry (GC-MS) analysis of the residues every fourth week until no TTZ could be detected. The recovery for analysis of TTZ in soil was between 98 and 131%, and the quantification level was 0.002 mg kg(-1). After 56 d of incubation, 20 and 28% of the TTZ applied remained in the soil and seed treatments, respectively, with corresponding half-lives of 27 and 29 d. The microbial biomass initially decreased in the soil treatment but had recovered after 56 d. The active part of the biomass was not changed during the experimental time. Thus, with respect to dissipation of TTZ and its effect on the soil microbial biomass and activity, no long-lasting difference between soil and seed treatments could be found.     

Degradation of biomacromolecules during high-rate composting of wheat straw-amended feces

Pig (Sus scrofa) feces, separately collected and amended with wheat straw, was composted in a tunnel reactor connected with a cooler. The composting process was monitored for 4 wk and the degradation of organic matter was studied by two chemical extraction methods, 13C cross polarization magic angle spinning (CPMAS) nuclear magnetic resonance (NMR) and pyrolysis gas chromatography-mass spectrometry (GC-MS). Wet-chemical extraction methods were not adequate to study the degradation of specific organic compounds as the extraction reagents did not give selective separation of hemicellulose, cellulose, proteins, and lignins. A new method was proposed to calculate the contribution of four biomacromolecules (aliphatics, proteins, polysaccharides, and lignin) from the 13C CPMAS NMR spectrum. Pyrolysis GC-MS allowed identification of the composition of the biomacromolecules. The biomacromolecules showed different rates of degradation during composting. High initial degradation rates of aliphatics, hemicellulose, and proteins were observed, where aliphatics were completely degraded and hemicellulose and proteins were partly recalcitrant during the four weeks of composting. The degradation rate of cellulose was much lower and degradation was not completed within the four weeks of composting. Lignin was not degraded during the thermophilic stage of composting but started to degrade slowly during the mesophilic stage. A combination of 13C CPMAS NMR and pyrolysis GC-MS gave good qualitative and semiquantitative assessments of the degradation of biomacromolecules during composting.     

SOIL SOLUTION CONCENTRATIONS: EFFECT OF EXTRACTION TIME USING POROUS CERAMIC CUPS UNDER CONSTANT TENSION1

ABSTRACT: Porous ceramic cups under constant tension (0.45 bar) were used to extract solutions from undisturbed soil columns. Solution concentration changed with length of extraction time. Significant relationships were found between extraction time and concentrations of P, Ca, and K in soil solution for two sample depths in an Omega loamy sand soil column. At two extraction time classes and at two sample depths, combined data from 12 soil columns representing two soil series reinforce the relationship. As time to extract a sample increases, the sample probably represents solution held by soil at tensions approaching those applied to the ceramic cup. We recommend that the choice of an extraction tension be given consideration in studies using porous ceramic cups under constant tension for monitoring constituents of soil solution. In addition, care must be taken to attain good physical contact between the cups and the soil material.     

Assessment of Endocrine‐Disrupting Chemicals Attenuation in a Coastal Plain Stream Prior to Wastewater Treatment Plant Closure

The U.S. Geological Survey is conducting a combined pre/post‐closure assessment at a long‐term wastewater treatment plant (WWTP) site at Fort Gordon near Augusta, Georgia. Here, we assess select endocrine‐active chemicals and benthic macroinvertebrate community structure prior to closure of the WWTP. Substantial downstream transport and limited instream attenuation of endocrine‐disrupting chemicals (EDCs) was observed in Spirit Creek over a 2.2‐km stream segment downstream of the WWTP outfall. A modest decline (less than 20% in all cases) in surface water detections was observed with increasing distance downstream of the WWTP and attributed to partitioning to the sediment. Estrogens detected in surface water in this study included estrone (E1), 17β‐estradiol (E2), and estriol (E3). The 5 ng/l and higher mean estrogen concentrations observed in downstream locations indicated that the potential for endocrine disruption was substantial. Concentrations of alkylphenol ethoxylate (APE) metabolite EDCs also remained statistically elevated above levels observed at the upstream control site. Wastewater‐derived pharmaceutical and APE metabolites were detected in the outflow of Spirit Lake, indicating the potential for EDC transport to aquatic ecosystems downstream of Fort Gordon. The results indicate substantial EDC occurrence, downstream transport, and persistence under continuous supply conditions and provide a baseline for a rare evaluation of ecosystem response to WWTP closure.     

Occurrence and fate of pesticides in four contrasting agricultural settings in the United States

Occurrence and fate of 45 pesticides and 40 pesticide degradates were investigated in four contrasting agricultural settings--in Maryland, Nebraska, California, and Washington. Primary crops included corn at all sites, soybeans in Maryland, orchards in California and Washington, and vineyards in Washington. Pesticides and pesticide degradates detected in water samples from all four areas were predominantly from two classes of herbicides--triazines and chloroacetanilides; insecticides and fungicides were not present in the shallow ground water. In most samples, pesticide degradates greatly exceeded the concentrations of parent pesticide. In samples from Nebraska, the parent pesticide atrazine [6-chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4-diamine] was about the same concentration as the degradate, but in samples from Maryland and California atrazine concentrations were substantially smaller than its degradate. Simazine [6-chloro-N,N'-diethyl-1,3,5-triazine-2,4-diamine], the second most detected triazine, was detected in ground water from Maryland, California, and Washington. Metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl)acetamide] rarely was detected without its degradates, and when they were detected in the same sample metolachlor always had smaller concentrations. The Root-Zone Water-Quality Model was used to examine the occurrence and fate of metolachlor at the Maryland site. Simulations accurately predicted which metolachlor degradate would be predominant in the unsaturated zone. In analyses of relations among redox indicators and pesticide variance, apparent age, concentrations of dissolved oxygen, and excess nitrogen gas (from denitrification) were important indicators of the presence and concentration of pesticides in these ground water systems.