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Xenoestrogen-induced ERK-1 and ERK-2 activation via multiple membrane-initiated signaling pathways.

Xenoestrogens can mimic or antagonize the activity of physiological estrogens, and the suggested mechanism of xenoestrogen action involves binding to estrogen receptors (ERs). However, the failure of various in vitro or in vivo assays to show strong genomic activity of xenoestrogens compared with estradiol (E2) makes it difficult to explain their ability to cause abnormalities in animal (and perhaps human) reproductive functions via this pathway of steroid action. E2 has also been shown to initiate rapid intracellular signaling, such as changes in levels of intracellular calcium, cAMP, and nitric oxide, and activations of a variety of kinases, via action at the membrane. In this study, we demonstrate that several xenoestrogens can rapidly activate extracellular-regulated kinases (ERKs) in the pituitary tumor cell line GH3/B6/F10, which expresses high levels of the membrane receptor for ER-alpha (mER). We tested a phytoestrogen (coumestrol), organochlorine pesticides or their metabolites (endosulfan, dieldrin, and DDE), and detergent by-products of plastics manufacturing (p-nonylphenol and bisphenol A). These xenoestrogens (except bisphenolA) produced rapid (3-30 min after application), concentration (10(-14)-10(-8) M)-dependent ERK-1/2 phosphorylation but with distinctly different activation patterns. To identify signaling pathways involved in ERK activation, we used specific inhibitors of ERs, epidermal growth factor receptors, Ca2+ signaling, Src and phosphoinositide-3 kinases, and a membrane structure disruption agent. Multiple inhibitors blocked ERK activation, suggesting simultaneous use of multiple pathways and complex signaling web interactions. However, inhibitors differentially affected each xenoestrogen response examined. These actions may help to explain the distinct abilities of xenoestrogens to disrupt reproductive functions at low concentrations.

Animals↗

Comparison of the promoting effects of various agents in induction of preneoplastic lesions in rat liver.

The promoting activities of 29 compounds on induction of hyperplastic (neoplastic) liver nodules (HN) and the dose-dependent effects of tumor promoting agents were compared by using a short-term test system developed in this laboratory. In tests on promoting activity, F344 rats were given a single dose (200 mg/kg) of N-nitrosodiethylamine (DEN), and from two weeks later, were treated with various test compounds for 6 or 10 weeks. They were subjected to partial hepatectomy 3 or 4 weeks after DEN treatment. The results showed that strong hepatocarcinogens, such as aflatoxin B1, DEN, N-nitrosodimethylamine (DMN), 2-acetylaminofluorene (2-AAF), 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) and ethionine, induced many hyperplastic liver nodules, whereas dieldrin, 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT), polychlorinated biphenyls (PCB) and alpha-hexachlorocyclohexane (alpha-HCH) induced few lesions. Nonhepatocarcinogens, such as N-nitrosoethylurea (ENU) and 3-methylcholanthrene (3-MC), only slightly induced hyperplastic nodules. Of the miscellaneous compounds tested, phenobarbital, deoxycholic acid and ethynyl estradiol also induced gamma-glutamyl transpeptidase (gamma-GT) positive foci. In tests on the dose-dependent effects of promoting agents, DMN was given at different concentrations for 6 weeks from 2 weeks after DEN treatment. Results were quantitated by histochemical measurement of the number or area of gamma-GT positive lesions induced. A long-term experiment on the effect of feeding DMN for 96 weeks was also done. Clear dose-dependent effects of DMN were seen in induction of gamma-GT positive foci in the short-term experiment and neoplastic lesions in the long-term one.

2-Acetylaminofluorene↗

Association of in utero organochlorine pesticide exposure and fetal growth and length of gestation in an agricultural population.

From 1940 through the 1970s, organochlorine compounds were widely used as insecticides in the United States. Thereafter, their use was severely restricted after recognition of their persistence in the environment, their toxicity in animals, and their potential for endocrine disruption. Although substantial evidence exists for the fetal toxicity of organochlorines in animals, information on human reproductive effects is conflicting. We investigated whether infants' length of gestation, birth weight, and crown-heel length were associated with maternal serum levels of 11 different organochlorine pesticides: p,p -dichlorodiphenyltrichloroethane (p,p -DDT), p,p -dichlorodiphenyldichloroethylene (p,p -DDE), o,p -dichlorodiphenyltrichloroethane (o,p -DDT), hexachlorobenzene (HCB), gamma-hexachlorocyclohexane (gamma-HCCH), gamma-hexachlorocyclohexane (gamma-HCCH), dieldrin, heptachlor epoxide, oxychlordane, trans-nonachlor, and mirex. Our subjects were a birth cohort of 385 low-income Latinas living in the Salinas Valley, an agricultural community in California. We observed no adverse associations between maternal serum organochlorine levels and birth weight or crown-heel length. We found decreased length of gestation with increasing levels of lipid-adjusted HCB (adjusted gamma = -0.47 weeks; p = 0.05). We did not find reductions in gestational duration associated with any of the other organochlorine pesticides. Our finding of decreased length of gestation related to HCB does not seem to have had clinical implications for this population, given its relatively low rate of preterm delivery (6.5%).

Agriculture↗

Relationships between toxicopathic hepatic lesions and exposure to chemical contaminants in English sole (Pleuronectes vetulus), starry flounder (Platichthys stellatus), and white croaker (Genyonemus lineatus) from selected marine sites on the Pacific Coast, USA.

Relationships between hepatic lesions and chemical contaminant concentrations in sediments, stomach contents, liver tissue, and bile were statistically evaluated in three species of bottomfish, English sole (Pleuronectes vetulus), starry flounder (Platichthys stellatus), and white croaker (Genyonemus lineatus), captured from 27 urban and nonurban sites on the Pacific Coast from Alaska to southern California. Lesions detected were neoplasms, preneoplastic foci of cellular alteration, nonneoplastic proliferative lesions, unique or specific degenerative/necrotic lesions, nonspecific degenerative/necrotic lesions, and hydropic vacuolation of biliary epithelial cells and hepatocytes. In general, lesion prevalences were significantly higher in all three species captured at chemically contaminated urban sites, and certain lesions had significantly higher relative risks of occurrence at urban sites in Puget Sound, San Francisco Bay, the vicinity of Los Angeles, and San Diego Bay. Concentrations of polycyclic aromatic hydrocarbons, polychlorinated biphenyls, DDT and its derivatives, and chlordanes and dieldrin in sediment, stomach contents, liver, and fluorescent aromatic compounds in bile were significant risk factors for the occurrence of neoplastic, preneoplastic, nonneoplastic proliferative, and specific degenerative/necrotic lesions, as well as hydropic vacuolation. Fish age also had a significant influence on occurrence of several hepatic lesions, but gender was rarely a significant risk factor. These relationships provide strong evidence for the involvement of environmental contaminants in the etiology of hepatic lesions in several marine bottomfish species and clearly indicate the utility of these lesions as biomarkers of contaminant-induced effects in wild fish.

Animals↗

Chemical contaminants, pharmacokinetics, and the lactating mother.

We review the commonly occurring persistent pesticides and industrial chemicals in breast milk. These chemicals are dichlorodiphenyl trichloroethane as dichlorodiphenyl dichloroethene dieldrin, chlordane as oxychlordane, heptachlor, polychlorinated biphenyls, polychlorinated dibenzofurans, and polychlorinated dibenzodioxins. We present a worked example of the kinds of pharmacokinetic assumptions and calculations necessary for setting regulatory limits of contaminants in the food supply, calculating dose of chemical contaminants to the nursed infant, converting risks from lifetime exposure in laboratory animals to risks for short-term exposure in humans, and estimating the excess cancer risk to the nursed infant.

Animals↗

Chlorinated organic contaminants in breast milk of New Zealand women.

Breast milk samples from 38 women in New Zealand were analyzed for organochlorine pesticides, polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) as part of a World Health Organization collaborative study of breast-milk contaminants. The women were recruited from two urban areas (Auckland and Christchurch) and two rural areas (Northland and North Canterbury) in the North and South Islands of New Zealand. The best predictor of contaminant concentrations in breast milk was found to be the age of the mother. Regional differences were found for hexachlorobenzene, dieldrin, and pp-DDE, reflecting historical use patterns. Urban-rural differences were found for several PCBs, PCDDs, and PCDFs when contaminant concentrations were calculated on a whole-milk basis. However, these differences could be attributed to variation in breast-milk fat concentrations between urban and rural mothers. Urban mothers had about 50% more breast-milk fat than rural mothers. Evidence suggests that breast-milk consumption by babies is regulated by caloric intake. Almost all of the caloric content of milk is in the fat fraction. This suggests that breast-milk contaminant levels calculated on a whole-milk basis do not necessarily reflect the relative levels of exposure of infants to these contaminants. However, the factors that influence breast-milk fat concentration deserve further study.

Adult↗

The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants.

Estrogens are defined by their ability to induce the proliferation of cells of the female genital tract. The wide chemical diversity of estrogenic compounds precludes an accurate prediction of estrogenic activity on the basis of chemical structure. Rodent bioassays are not suited for the large-scale screening of chemicals before their release into the environment because of their cost, complexity, and ethical concerns. The E-SCREEN assay was developed to assess the estrogenicity of environmental chemicals using the proliferative effect of estrogens on their target cells as an end point. This quantitative assay compares the cell number achieved by similar inocula of MCF-7 cells in the absence of estrogens (negative control) and in the presence of 17 beta-estradiol (positive control) and a range of concentrations of chemicals suspected to be estrogenic. Among the compounds tested, several "new" estrogens were found; alkylphenols, phthalates, some PCB congeners and hydroxylated PCBs, and the insecticides dieldrin, endosulfan, and toxaphene were estrogenic by the E-SCREEN assay. In addition, these compounds competed with estradiol for binding to the estrogen receptor and increased the levels of progesterone receptor and pS2 in MCF-7 cells, as expected from estrogen mimics. Recombinant human growth factors (bFGF, EGF, IGF-1) and insulin did not increase in cell yields. The aims of the work summarized in this paper were a) to validate the E-SCREEN assay; b) to screen a variety of chemicals present in the environment to identify those that may be causing reproductive effects in wildlife and humans; c) to assess whether environmental estrogens may act cumulatively; and finally d) to discuss the reliability of this and other assays to screen chemicals for their estrogenicity before they are released into the environment.

Animals↗

Behavioral toxicology, risk assessment, and chlorinated hydrocarbons.

Behavioral end points are being used with greater frequency in neurotoxicology to detect and characterize the adverse effects of chemicals on the nervous system. Behavioral measures are particularly important for neurotoxicity risk assessment since many known neurotoxicants do not result in neuropathology. The chlorinated hydrocarbon class consists of a wide variety of chemicals including polychlorinated biphenyls, clioquinol, trichloroethylene, hexachlorophene, organochlorine insecticides (DDT, dicofol, chlordecone,dieldrin, and lindane), and phenoxyherbicides. Each of these chemicals has effects on motor, sensory, or cognitive function that are detectable using functional measures such as behavior. Furthermore, there is evidence that if exposure occurs during critical periods of development, many of the chlorinated hydrocarbons are developmental neurotoxicants. Developmental neurotoxicity is frequently expressed as alterations in motor function or cognitive abilities or changes in the ontogeny of sensorimotor reflexes. Neurotoxicity risk assessment should include assessments of the full range of possible neurotoxicological effects, including both structural and functional indicators of neurotoxicity.

Behavior↗

Organochlorines in breast milk from two cities in Ukraine.

Reports of environmental problems in the former Soviet Union, including excess use of pesticides, have led to concerns about high levels of contamination in humans, but little information is available to assess whether these concerns are warranted. Samples of breast milk from 197 women from two cities in Ukraine were analyzed for p,p'-DDT, p,p'-DDE, endrin, dieldrin, heptachlor epoxide, trans-nonachlor, oxychlordane, hexachlorobenzene, ss-hexachlorocyclohexane (HCH), and 18 polychlorinated biphenyl congeners, and results were compared to previous reports from Europe. The median ss-HCH concentration was 731 ng/g milk fat, which is higher than other reports from Europe but lower than reports from other parts of the world. The median DDE concentration was 2,457 ng/g milk fat, which is higher than most but not all other reports from Europe. Concentrations of other chemicals were comparable to or lower than other reports from Europe. Concentrations from the city of Kyiv were generally lower than those from Dniprodzerzhinsk, but the magnitudes of these differences were modest.

Adult↗

Persistent organochlorine pollutants in children working at a waste-disposal site and in young females with high fish consumption in Managua, Nicaragua.

The aim of this study was to assess persistent organochlorine pollutant (POP) levels in serum collected from children (11-15 years old) working and sometimes also living at the municipal waste-disposal site in Managua, located at the shore of Lake Managua, and in nonworking children living both nearby and also far away from the waste-disposal site. The influence of fish consumption was further evaluated by assessing POPs levels in serum from young women (15-24 years old) with markedly different patterns of fish consumption from Lake Managua. 2,2-bis(4-chlorophenyl)-1,1,1-trichloro-ethane (4,4'-DDT) and 2,2-bis(4-chlorophenyl)-1,1-dichloro-ethene (4,4'-DDE), gamma-hexachlorocyclohexane (gamma-HCH), polychlorinated biphenyls, pentachlorophenol, and polychlorobiphenylols were quantified in all samples. In general, the levels observed were higher than those reported in children from developed countries, such as Germany and United States. Toxaphene, aldrin, dieldrin, and beta-HCH could not be identified in any sample. The children working at the waste-disposal site had higher levels of POPs compared with the nonworking reference groups. In children not working, there were also gradients for several POPs, according to vicinity to the waste-disposal site. Moreover, in children, as well as in young women, there were gradients according to fish consumption. The most abundant component was 4,4'-DDE, but at levels still lower than those reported in children from malarious areas with a history of recent or current application of 4,4'-DDT for vector control.

Adolescent↗

[Levels of exposure to organochlorine pesticides in open-air dump dwellers].

OBJECTIVE: To estimate the degree of internal exposure to organochlorine pesticides (persistent organic pollutants, POP) among inhabitants of an open-air dump. METHODS: This was an observational study with simultaneous measurement and control, in which the criterion for subjects was that they dwelled in an area contaminated by POP and the effect of such contamination was the concentration of such substances into the blood. The study population consisted of 238 people living on an open-air dump in Pil es, in the municipality of Cubatão, SP, Brazil, and the control population was formed by 258 people, also resident in that municipality, in the Cota 200 district. The following pesticides were analysed: hexachlorobenzene (HCB), p-p'DDT, p-p'DDE, p-p'DDD, o-p'DDT, a HCH (hexachlorocyclohexane); b HCH; g HCH; aldrin; dieldrin; endrin; heptachlor; heptachlor-epoxy and mirex. RESULTS: The average blood level of HCB in Pil es was 4.66 g/l, which was 155 times higher than the average for Cota 200 (0.03 g/l). In Pil es the average blood concentration of total DDT was 3.71 g/l, which was twice the level found in Cota 200 (1.85 g/l). Total HCH presented blood concentrations that were six times greater in Pil es (0.84 g/l versus 0.13 g/l). CONCLUSIONS: A positive association was shown between dwelling in Pil es and the presence of POP in the blood, with much higher risks than in localities without the presence of these contaminants.

Adolescent↗

Serum levels of organochlorine pesticides and polychlorinated biphenyls among inhabitants of Greater Metropolitan Rio de Janeiro, Brazil.

Levels of persistent organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) were determined in the blood serum of people living and working in the urban area of greater Rio de Janeiro city. Blood samples from 33 volunteers (16 males, 17 females, 19-63 years old) were taken in January 1999. OCP residues (op'DDT, pp'DDT, pp'DDD, pp'DDE, Aldrin, Dieldrin, Endrin, Heptachlor, Heptachlor-epoxide, alpha-, beta- and gamma-Hexachlorocyclo-hexane, Hexachlorobenzene) and PCBs (congeners: 28, 52, 101, 138, 153, 180) were extracted with n-hexane and analyzed by gas chromatography with electron capture detection. Except for pp'DDE (detection limit = 1.4 microg/L) no other OCP residue was found in the samples. No PCB congener (detection limit = 2.0 microg/L) was detected either. pp'DDE was found in 17 out of 33 samples in concentrations that ranged from 1.4 to 8.4 microg/L of serum or, on a fat basis, from 0.200 to 3.452 microg/g of serum lipids. Percentage of positive samples (%) and levels of pp'DDE (range of positive samples) increased from the youngest to the oldest group (< or = 29 yrs: 10%, 0.278 microg/g; 30-39 yrs: 60%, 0.200-0.765 microg/g; > or = 40 yrs: 77%, 0.257-3.452 microg/g).

Adult↗

Two complementary bioassays for screening the estrogenic potency of xenobiotics: recombinant yeast for trout estrogen receptor and trout hepatocyte cultures.

A relation between the chemical structure of a xenobiotic and its steroidal action has not yet been clearly established. Thus, it is not possible to define the estrogenic potency of different xenobiotics. An assessment may be accomplished by the use of different bioassays. We have previously developed a yeast system highly and stably expressing rainbow trout estrogen receptor (rtER) in order to analyze the biological activity of the receptor. The recombinant yeast system appears to be a reliable, rapid and sensitive bioassay for the screening and determination of the direct interaction between ER and estrogenic compounds. This system was used in parallel with a more elaborate biological system, trout hepatocyte aggregate cultures, to examine the estrogenic potency of a wide spectrum of chemicals commonly found in the environment. In hepatocyte cultures, the vitellogenin gene whose expression is principally dependent upon estradiol was used as a biomarker. Moreover, competitive binding assays were performed to determine direct interaction between rtER and xenobiotics. In our study, 50% of the 49 chemical compounds tested exhibited estrogenic activity in the two bioassays: the herbicide diclofop-methyl; the fungicides biphenyl, dodemorph, and triadimefon; the insecticides lindane, methyl parathion, chlordecone, dieldrin, and endosulfan; polychlorinated biphenyl mixtures; the plasticizers or detergents alkylphenols and phthalates; and phytoestrogens. To investigate further biphenyl estrogenic activity, its principal metabolites were also tested in both bioassays. Among these estrogenic compounds, 70% were able to activate rtER in yeast and hepatocytes with variable induction levels according to the system. Nevertheless, 30% of these estrogenic compounds exhibited estrogenic activity in only one of the bioassays, suggesting the implication of metabolites or different pathways in the activation of gene transcription. This paper shows that it is important to combine in vivo bioassays with in vitro approaches to elucidate the mechanism of xenoestrogen actions.

Animals↗

Micronucleus induction in gill cells of green-lipped mussels (Perna viridis) exposed to mixtures of polycyclic aromatic hydrocarbons and chlorinated pesticides.

Different groups of green-lipped mussels (Perna viridis) were exposed to the same net amount of a genotoxicant mixture of four polycyclic aromatic hydrocarbons ([PAHs]; anthracene, fluoranthene, pyrene, and benzo[a]pyrene) and four organochlorine pesticides ([OCs]; alpha-hexachlorocyclohexane (HCH), aldrin, dieldrin, and p,p'-DDT) for four weeks under different regimes that simulated various scenarios of fluctuating toxicant levels in the marine environment. Micronucleus (MN) formation in gill cells was studied at the end of each week. Micronucleus frequencies increased with continual addition of genotoxicants, and did not diminish significantly under conditions of either gradually decreasing concentrations or cessation of exposure for one to two weeks, suggesting that the MN response may persist over relatively long exposure periods. An almost two-fold higher mean MN frequency was recorded in a chronic exposure group than in an acute group that had received the same net nominal dose of genotoxicants, indicating that chronic exposure may lead to a greater genotoxic impact than acute exposure. The results suggested that in field studies, MN response should be monitored at multiple time points in order to elucidate the effects of potentially fluctuating toxicant levels. Finally, MN formation was positively correlated with both nominal contaminant levels and tissue levels of the genotoxicants. These findings suggest that MN responses can be a sensitive indicator of exposure to relatively low levels of genotoxicants and that MN response in mussel gill cells can be a stable biomarker of genotoxicity.

Animals↗

Factors influencing persistent organic pollutant concentrations in the Atlantic white-sided dolphin (Lagenorhynchus acutus).

Assessing the trends of persistent organic pollutants (POPs) in cetaceans is difficult because of age and gender influences on accumulation. Persistent organic pollutants bioaccumulate and are poorly metabolized; hence, concentrations may increase with age in males while females reduce their POP burden through parturition and lactation. Age and gender effects on contaminant concentrations are species specific because of life history and reproductive strategies. These influences must be understood in order to elucidate and assess lifetime POP exposure. The objectives of this study were to determine baseline POP concentrations in blubber samples from the Atlantic white-sided dolphin (Lagenorhynchus acutus) and to investigate life history and other influences, such as metabolism, on these concentrations. Forty-seven L. acutus blubber samples collected from mass stranding events in Massachusetts, USA (1993-2000), and archived in the National Marine Mammal Tissue Bank at the National Institute of Standards and Technology (Gaithersburg, MD, USA) were analyzed for 55 polychlorinated biphenyl congeners (PCB; 55 congeners), five polybrominated diphenyl ether (PBDE) congeners, and organochlorine pesticides (toxaphene, DDT and metabolites, mirex, dieldrin, chlordanes, hexachlorocyclohexanes, hexachlorobenzene, and endosulfans) by gas chromatography/mass spectrometry (GC-MS). Ages for 19 animals were determined from growth layer groups on decalcified, stained thin tooth sections. Total PCBs (sigmaPCB; sum of 55 congeners) were the contaminants present in the highest concentrations in all age classes (0.5-63 microg/g wet mass) followed by sum of DDTs (0.50-43 microg/g wet mass), toxaphene (0.055-31 microg/g wet mass), chlordanes (0.30-24 microg/g wet mass), and PBDEs (0.12-4.0 microg/g wet mass). Body length had a greater statistical influence than age on contaminant burdens in L. acutus. Contaminant burdens decreased with length in both male and female L. acutus, suggesting metabolic elimination and/or growth dilution in males and off-loading via lactation in females.

Adipose Tissue↗

Are organohalogen contaminants a cofactor in the development of renal lesions in east Greenland polar bears (Ursus maritimus)?

Tissues of polar bears (Ursus maritimus) from East Greenland contain the highest concentrations of organohalogen contaminants (OHCs) among subpopulations of any mammalian species in the Arctic. Negative associations also have been found between OHC concentrations and bone mineral density and liver histology parameters for this subpopulation of polar bears. The present study examined the OHC concentrations and adverse effects on renal tissue for 75 polar bears collected during 1999 to 2002. Specific lesions were diffuse glomerular capillary wall thickening, mesangial glomerular deposits, tubular epithelial cell hyperplasia, hyalinization of the tubular basement membrane, tubular dilatation, atrophy and necrosis, tubular medullary hyalin casts, interstitial fibrosis, and mononuclear cell infiltration. With the exception of mononuclear cell infiltrations, all these parameters were correlated with age, whereas none was associated with the sex of the animals. In an age-controlled statistical analysis of covariance, increases in glomerular mesangial deposits and interstitial fibrosis were significantly (p < 0.05) correlated with polybrominated diphenyl ether (sigmaPBDE) concentrations in subadults. In adult males, statistically significant (p < 0.05) positive correlations were found for tubular epithelial cell hyperplasia and dieldrin concentration; diffuse glomerular capillary wall thickening and chlordane (sigmaCHL) concentrations, and tubular medullary hyalin casts and sigmaCHL, sigmaPBDE, polychlorinated biphenyl, and hexachlorocyclohexane concentrations. The lesions were consistent with those reported previously in highly OHC-contaminated Baltic seal populations and exposed laboratory animals. The renal lesions were a result of aging. However, based on the above statistical findings as well as the nature of the findings, we suggest that long-term exposure to OHCs may be a cofactor in renal lesion occurrence, although other cofactors, such as exposure to heavy metals and recurrent infections from microorganisms, cannot be ruled out. This is new and important knowledge in the assessment of health status among wildlife populations and humans relying on food resources that are contaminated with OHCs.

Animals↗

Sediment chemical contamination and toxicity associated with a coastal golf course complex.

The increasing density of golf courses represents a potential source of sediment contamination to nearby coastal areas, the chemical and biological magnitude of which is almost unknown. The objective of this study was to determine the concentrations of contaminants and toxicities of sediments impacted by a coastal golf course complex. Sediment contaminant concentrations were determined at least twice during the two-year study period at 14 sampling stations. In addition, a combination of acute and chronic bioassays were conducted exposing four invertebrate test species to whole sediments and associated pore waters. Overall, the Florida, USA, golf course complex had a measurable impact on sediment chemical quality, particularly in near-field areas. Higher concentrations of several trace metals and organochlorine pesticides were detected in many golf course-associated sediments compared with reference areas; however, concentrations decreased seaward and only a few, primarily chlorinated pesticides, exceeded proposed sediment quality guidelines. Chromium, zinc, and mercury were detected more frequently than other trace metals. The DDT and associated metabolites, dieldrin and chlordane, were the more commonly detected organic contaminants. Acute toxicity was uncommon and occurred consistently for sediment collected from one coastal location. In contrast, chronic toxicity occurred at several study sites based on the response of Mysidopsis bahia. It was concluded that the impact of golf course runoff on sediment quality may be subtle and sensitive biological assessment methods, such as chronic toxicity tests, will be needed to detect adverse effects.

Animals↗

Screening of environmental contaminants for ecdysteroid agonist and antagonist activity using the Drosophila melanogaster B(II) cell in vitro assay.

The B(II) bioassay was developed as a rapid and reliable tool for detecting potential insect growth regulators acting as ecdysteroid receptor (ant)agonists. Based on an ecdysteroid-responsive cell line from Drosophila melanogaster, this microplate assay is ideally suited to the evaluation of environmental contaminants as potential endocrine disrupters. Data are presented for about 80 potential environmental contaminants, including industrial chemicals, pesticides, pharmaceuticals, phytoestrogens, and vertebrate steroids, and are compared with data for known (ant)agonists. Apart from androst-4-ene-3,17-dione (a weak antagonist), vertebrate steroids were inactive at concentrations up to 10(-3) M. The vast majority of xenobiotics also showed no (ant)agonist activity. Among the industrial chemicals, antagonistic activity was observed for bisphenol A median effective concentration (EC50) of 1.0 x 10(-4) M and diethylphthalate (EC50 of 2.0 x 10(-3) M). Some organochlorine compounds also showed weak antagonistic activity, including o,p'-dichlorodiphenyldichloroethylene (DDE), p,p'-DDE, dieldrin, and lindane (EC50 of 3.0 x 10(-5) M). For lindane, bisphenol A, and diethylphthalate, activity is not associated with impurities in the samples and, for lindane and bisphenol A at least, the compounds are able to compete with ecdysteroids for the ligand binding site on the receptor complex, albeit at concentrations very much higher than those found in the environment. The only pharmaceutical showing any detectable antagonist activity was 17alpha-ethynylestradiol. In the context of recent publications on potential endocrine disruption in marine and freshwater arthropods, these findings suggest that, for some compounds (e.g., diethylstilbestrol), ecdysteroid receptor-mediated responses are unlikely to be involved in producing chronic effects. The B(II) assay has a potentially valuable role to play in distinguishing between endocrine-mediated, which normally occur at submicromolar concentrations, and pharmacological effects in insects and crustaceans.

Animals↗