Interpretation of fish biomarker data for identification,classification, risk assessment and testing of endocrine disrupting chemicals

Chemical induced changes in fish biomarkers vitellogenin (VTG), secondary sex characteristics (SSC), and sex ratio indicate modes/mechanisms of action (MOAs) of EAS (estrogen, androgen and steroidogenesis) pathways. These biomarkers could be used for defining MOAs and the causal link between MOAs and adverse effects in fish for the identification of endocrine disrupting chemicals (EDCs). This paper compiled data sets of 150 chemicals for VTG, 57 chemicals for SSC and 38 chemicals for sex ratio in fathead minnow, medaka and zebrafish. It showed 1) changes in fish biomarkers can indicate the MOAs as anticipated; 2) in addition to EAS pathways, chemicals with non-EAS pathways induced changes in fish biomarkers; 3) responses of fish biomarkers did not always follow the anticipated patterns of EAS pathways. These responses may result from the interaction of chemical-induced multiple MOAs and confounding factors like fish diet, infection, culture conditions, general toxicity and stress response. The complex response of fish biomarkers to a chemical of interest requires EDC testing at multiple biological levels. Interpretation of fish biomarker data should be combined with relevant information at different biological levels, which is critical for defining chemical specific MOAs. The utility of fish biomarker data for identification, classification, PBT assessment, risk assessment, and testing of EDCs in the regulatory context was discussed. This paper emphasizes the importance of fish biomarker data in the regulatory context, a weight of evidence approach for the interpretation of fish biomarker data and the need for defining levels of evidence for the identification of EDCs.     

The unpredictable effects of mixtures of androgenic and estrogenic chemicals on fish early life

Estuarine shallow areas and coastal lagoons are known to receive and concentrate multiple inputs, either from land, rivers or coastal areas, being intensively impacted by chemical contamination, namely endocrine disrupting chemicals (EDCs). Despite the ubiquitous coexistence of several classes of EDCs in most of these aquatic ecosystems, there is still limited information regarding their combined effects. Furthermore, given the immediate implications for population dynamics, the available laboratory studies almost invariably focus on very specific life history stages, such as embryonic development or reproduction, thus creating a gap on our knowledge of what happens in between. During this 'intermediate phase', the newborn larvae and juveniles face numerous challenges whose outcome may impair reproduction or even survival. The black-striped pipefish, Syngnathus abaster, member of the Syngnathidae family (comprising pipefish, seahorses and seadragons), usually breeds in coastal areas such as estuaries, where its newborns are immediately exposed to EDCs. Given the ongoing decline of pipefish populations, together with the observed shrinkage and fragmentation of seagrass meadows, known to be impacted by EDCs, a first reasonable question to address is if pipefish newborns respond to environmentally relevant concentrations of ubiquitous EDCs, either single or in combination. Hence, a seven days exposure experiment to the estrogenic chemical ethinylestradiol (EE(2)) and the androgenic chemical tributyltin (TBT), single and in binary mixtures, was conducted. Selected behavioural (e.g. predator avoidance) and developmental variables (e.g. growth) were monitored in pipefish juveniles after EDCs insult. The obtained results indicate that EE(2), TBT, or their combined exposure, do impact pipefish early life. However, the pattern of results emerging from the measured variables clearly indicates that mixtures significantly modulate newborn responses in distinct ways when compared to individual chemical's exposure. These findings further demonstrate the importance of addressing the issue of chemical mixtures of pollutants acting through dissimilar mode of action. Independently of all the observed response variations, an ultimate conclusion seems certain: EE(2) and TBT, single or in combination, induce disruption patterns able to imbalance pipefish survival. Since these (as well as other) contaminants are present in estuarine areas, profound implications in population structure could be expected, ranging from a decrease in recruitment to a disruption of sexual selection. Inexorably, these stressors simultaneously operate in already declining populations.     

Human exposure to endocrine disrupting chemicals and fertility: A case–control study in male subfertility patients

BackgroundDioxins, PCBs, chlorinated pesticides, brominated flame retardants, bisphenol A, triclosan, perfluorinated compounds and phthalates are known as endocrine disrupting chemicals (EDCs).ObjectivesThe aim of our study was to investigate whether higher exposure to EDCs is associated with increased subfertility in men.MethodsWe measured biomarkers of exposure in 163 men, recruited through four fertility clinics. According to WHO guidelines, we used a total motility count (TMC) of 20 million as cut-off value. We assigned patients to the case group when two semen samples – collected at least one week apart – had a TMC < 20 and to the control group when both samples had a TMC ≥ 20. To estimate the risk of subfertility and alteration in sex hormone concentrations we used multivariable-adjusted analysis, using logistic and linear regressions, respectively.ResultsFor an IQR increase in serum oxychlordane, the odds ratio for subfertility was 1.98 (95% CI: 1.07; 3.69). Furthermore, men with serum levels of BDE209 above the quantification limit had an odds of 7.22 (1.03; 50.6) for subfertility compared with those having values below the LOQ. Urinary levels of phthalates and triclosan were negatively associated with inhibin B and positively with LH. Urinary bisphenol A correlated negatively with testosterone levels.ConclusionsOur study in men showed that internal body concentrations of endocrine disrupting chemicals are associated with an increased risk of subfertility together with alterations in hormone levels. The results emphasize the importance to reduce chemicals in the environment in order to safeguard male fertility.     

Screening of hormone-like activities in bottled waters available in Southern Spain using receptor-specific bioassays

Bottled water consumption is a putative source of human exposure to endocrine-disrupting chemicals (EDCs). Research has been conducted on the presence of chemicals with estrogen-like activity in bottled waters and on their estrogenicity, but few data are available on the presence of hormonal activities associated with other nuclear receptors (NRs). The aim of this study was to determine the presence of endocrine activities dependent on the activation of human estrogen receptor alpha (hERa) and/or androgen receptor (hAR) in water in glass or plastic bottles sold to consumers in Southern Spain. Hormone-like activities were evaluated in 29 bottled waters using receptor-specific bioassays based on reporter gene expression in PALM cells [(anti-)androgenicity] and cell proliferation assessment in MCF-7 cells [(anti-)estrogenicity] after optimized solid phase extraction (SPE). All of the water samples analyzed showed hormonal activity. This was estrogenic in 79.3% and anti-estrogenic in 37.9% of samples and was androgenic in 27.5% and anti-androgenic in 41.3%, with mean concentrations per liter of 0.113 pM 17β-estradiol (E₂) equivalent units (E₂Eq), 11.01 pM anti-estrogen (ICI 182780) equivalent units (ICI 182780Eq), 0.33 pM methyltrienolone (R1881) equivalent units (R1881Eq), and 0.18 nM procymidone equivalent units (ProcEq). Bottled water consumption contributes to EDC exposure. Hormone-like activities observed in waters from both plastic and glass bottles suggest that plastic packaging is not the sole source of contamination and that the source of the water and bottling process may play a role, among other factors. Further research is warranted on the cumulative effects of long-term exposure to low doses of EDCs.     

Natural and synthetic endocrine disrupting compounds (EDCs) in water, sediment and biota of a coastal lagoon

We report a survey on the occurrence and distribution of natural (17beta-estradiol, E2; estrone, E1) and synthetic (nonylphenol, NP; nonylphenol monoethoxylate carboxylate, NP1EC; bisphenol-A, BPA; benzophenone, BP; mestranol, MES; 17alpha-ethinylestradiol, EE2; diethylstilbestrol, DES) endocrine disrupting compounds (EDCs) in water, sediment and biota (Mediterranean mussel, Mytilus galloprovincialis) in the Venice lagoon, a highly urbanized coastal water ecosystem that receives both industrial and municipal wastewater effluents. The survey was preceded by the development of tailor made extraction and clean-up procedures for the simultaneous HPLC-ESI-MS determination of all examined EDCs in sediment and biota samples. Satisfactory extraction performances and method detection limits (MDLs) were obtained for almost all EDCs. Most of the selected compounds were found in water and sediment (concentration range: 2.8-211 ng/L, and 3.1-289 microg/kg, d.w., respectively), while only 17alpha-ethinylestradiol and nonylphenol were recorded in biota samples (conc. range: 7.2-240 ng/g, d.w.). 17beta-estradiol and ethinylestradiol contributed mostly to the water estradiol equivalent concentration (EEQ) (1.1-191 ng/L, average: 25 ng/L), while synthetic EDCs (17alpha-ethinylestradiol, diethylstilbestrol) were mainly responsible of the sediment EEQ (1.1-191 microg/kg, average: 71 microg/kg, d.w.). Whenever diethylstilbestrol was not recorded in the sediment, water EEQs were similar to sediment EEQs. A remarkable increase of nonylphenol was observed in sediments over the last decade.     

Estuarine and coastal zone marine pollution by the nonionic alkylphenol ethoxylates endocrine disrupters: is there a potential ecotoxicological problem?

The nonionic biodegradation-resistant ("hard") alkylphenol ethoxylate (APEO) surfactants and their degradation products are known endocrine disrupting chemicals (EDCs). We report here the findings concerning the APEOs concentrations and homologic distribution profiles in Israel's estuarine and coastal zone seawater to serve as a case study. The concentrations in sewage-containing rivers, estuaries and 50-60-m offshore sea (Mediterranean) water were found to be 12.5-75.1, 4.2-25.0 and 0.9-2.6 microg/L, respectively. The corresponding homologic distribution profiles were found to be within the range of 1-10% each, somewhat skewing, as expected, towards the more toxic shorter-chain ethoxylates. Egg production by zebrafish, exposed to 75, 25 and 10 microg/L of a typical industrial APEOs was reduced up to 89.6%, 84.7% and 76.9%, respectively, between the 8th and 28th days of exposure. Apparently, there is a potential APEOs-related ecotoxicological health risk problem.     

Removal of trace organic contaminants from domestic wastewater: A meta-analysis comparison of sewage treatment technologies

Trace organic contaminants (TrOCs), such as endocrine disrupting compounds (EDCs) and pharmaceuticals and personal care products (PPCPs), represent global threats to aquatic animals and ecosystems. A major source of TrOCs in the aquatic environment is via the discharge of treated sewage, so there is an urgent need to evaluate the comparative efficiencies of the most widely used sewage treatment technologies as regards elimination of these compounds from wastewater. To address this need, 976 published articles were compiled focusing on estimates of removal (%) for 20 common environmental TrOCs, from five major sewage treatment technologies: conventional activated sludge (CAS), oxidation ditch (OD), membrane bioreactor (MBR), ponds and constructed wetlands (PCW), and trickling biological filters (TBF). A quantitative meta-analysis was performed to compare standardized relative removal efficiencies (SREs) of the compounds amongst these technologies, and where possible potential sources of heterogeneity were considered (e.g., flow rates and chemical sorption potential). The results indicate that the most widely used CAS treatment and the less common TBF provide comparatively poor overall removal of common organic micropollutants. Membrane bioreactors appear to be capable of achieving the greatest overall removal efficiencies, but the sustainability and economic viability of this option has been questioned. Treatment with OD systems may be more economical while still achieving comparatively high removal efficiencies, and the analysis revealed OD to be the best option for targeting highly potent estrogenic EDCs. This study offers a unique global assessment of TrOC removal via leading sewage treatment technologies, and is an important step in the identification of effective options for treating municipal sewage.     

Exposure to endocrine disrupting chemicals among residents of a rural vegetarian/vegan community

Background & AimsEndocrine-disrupting chemicals (EDCs) are increasingly thought to be involved in the rising prevalence of disorders such as obesity, diabetes, and some hormone-dependent cancers. Several lines of evidence have indicated that vegetarian and vegan diets may offer some protection from such diseases. We hypothesized that exposure to selected EDCs among residents of the unique vegetarian/vegan community of Amirim would be lower than what has recently been reported for the omnivorous population in the first Israel Biomonitoring Study (IBMS).MethodsWe studied 42 Amirim residents (29 vegetarians/13 vegans; 24 women/18men, aged 50.7 ± 13.7 y). Subjects answered detailed lifestyle, and multipass, memory-based 24-hr dietary recall questionnaires. Concentrations of bisphenol A (BPA), 11 phthalate metabolites, and the isoflavone phytoestrogens (genistein and daidzein) were determined by GC or LC tandem mass-spectrometry on a spot urine sample. The results were compared to those obtained following the same methodology in the Jewish subgroup of the IBMS (n = 184).ResultsWhile a vegetarian/vegan nutritional pattern had no effect on exposure to BPA, it seemed to confer a modest protection (~ 21%) from exposure to high molecular weight phthalates. Furthermore, the summed metabolites of the high molecular weight phthalate DiNP were 36% lower in vegans compared to vegetarians (P < 0.05). In contrast, Amirim residents exhibited a level of exposure to isoflavone phytoestrogens about an order of magnitude higher than in the IBMS (P < 0.001).ConclusionsIn Israel, a country whose inhabitants demonstrate exposure to EDCs comparable to that of the US and Canada, a voluntary lifestyle of vegetarianism and preference for organic food has a modest, but possibly valuable, impact on exposure to phthalates, while it is associated with a very steep increase in the exposure to phytoestrogens. Major reduction in exposure to EDCs will require regulatory actions.     

A global assessment of phthalates burden and related links to health effects

Phthalates are ubiquitous environmental contaminants which are used in industry as plasticizers and additives in cosmetics. They are classified as Endocrine Disrupting Chemicals (EDCs) which impair the human endocrine system inducing fertility problems, respiratory diseases, childhood obesity and neuropsychological disorders. The aim of this review is to summarize the current state of knowledge on the toxicity that phthalates pose in humans based on human biomonitoring studies conducted over the last decade. Except for conventional biological matrices (such as urine and serum), amniotic fluid, human milk, semen, saliva, sweat, meconium and human hair are also employed for the estimation of exposure and distribution of pollutants in the human body, although data are not enough yet. Children are highly exposed to phthalates relative to adults and in most studies children's daily intake surpasses the maximum reference dose (RfD) set from US Environmental Protection Agency (US EPA). However, the global trend is that human exposure to phthalates is decreasing annually as a result of the strict regulations applied to phthalates.     

Chemical contaminants in swimming pools: Occurrence,implications and control

A range of trace chemical contaminants have been reported to occur in swimming pools. Current disinfection practices and monitoring of swimming pool water quality are aimed at preventing the spread of microbial infections and diseases. However, disinfection by-products (DBPs) are formed when the disinfectants used react with organic and inorganic matter in the pool. Additional chemicals may be present in swimming pools originating from anthropogenic sources (bodily excretions, lotions, cosmetics, etc.) or from the source water used where trace chemicals may already be present. DBPs have been the most widely investigated trace chemical contaminants, including trihalomethanes (THMs), haloacetic acids (HAAs), halobenzoquinones (HBQs), haloacetonitriles (HANs), halonitromethanes (HNMs), N-nitrosamines, nitrite, nitrates and chloramines. The presence and concentrations of these chemical contaminants are dependent upon several factors including the types of pools, types of disinfectants used, disinfectant dosages, bather loads, temperature and pH of swimming pool waters. Chemical constituents of personal care products (PCPs) such as parabens and ultraviolet (UV) filters from sunscreens have also been reported. By-products from reactions of these chemicals with disinfectants and UV irradiation have been reported and some may be more toxic than their parent compounds. There is evidence to suggest that exposure to some of these chemicals may lead to health risks. This paper provides a detailed review of various chemical contaminants reported in swimming pools. The concentrations of chemicals present in swimming pools may also provide an alternative indicator to swimming pool water quality, providing insights to contamination sources. Alternative treatment methods such as activated carbon filtration and advanced oxidation processes may be beneficial in improving swimming pool water quality.     

Quantitative determination of volatile organic compounds in indoor dust using gas chromatography-UV spectrometry

A novel technique, gas chromatography-UV spectrometry (GC-UV), was used to quantify volatile organic compounds (VOCs) in settled dust from 389 residences in Sweden. The dust samples were thermally desorbed in an inert atmosphere and evaporated compounds were concentrated by solid phase micro extraction and separated by capillary GC. Eluting compounds were then detected, identified, and quantified using a diode array UV spectrophotometer. Altogether, 28 compounds were quantified in each sample; 24 of these were found in more than 50% of the samples. The compounds found in highest concentrations were saturated aldehydes (C5-C10), furfuryl alcohol, 2,6-di-tert-butyl-4-methylphenol (BHT), 2-furaldehyde, and benzaldehyde. Alkenals were also found, notably 2-butenal (crotonaldehyde), 2-methyl-propenal (methacrolein), hexenal, heptenal, octenal, and nonenal. The concentrations of each of the 28 compounds ranged between two to three orders of magnitude, or even more. These results demonstrate the presence of a number of VOCs in indoor dust, and provide, for the first time, a quantitative determination of these compounds in a larger number of dust samples from residents. The findings also illustrate the potential use of GC-UV for analysing volatile compounds in indoor dust, some of which are potential irritants (to the skin, eyes or respiratory system) if present at higher concentrations. The potential use of GC-UV for improving survey and control of the human exposure to particle-bound irritants and other chemicals is inferred.     

Determination of parabens and benzophenone-type UV filters in human placenta. First description of the existence of benzyl paraben and benzophenone-4

UV filters and parabens (PBs) are chemicals used in daily personal care and hygiene products to protect materials and humans from the adverse effects of UV radiation and to preserve the integrity of the formulation, respectively. Several studies highlight their widespread environmental occurrence and endocrine disrupting effects. However, little is known about human exposure to these compounds. The objective of this study was to investigate the exposure of human embryos and foetuses to endocrine disrupting UV filters and PBs.Placentas from volunteer mothers in Barcelona were collected at delivery after informed, written consent by the pregnant women. UV filters and parabens were analysed by liquid chromatography-tandem mass spectrometry. The excellent performance of the method allowed measuring the target compounds in human placental tissue at low ng/g fresh weight level. The detection frequency of the selected compounds was in the range 17–100%. Benzophenone-1, methyl paraben, butyl paraben and benzyl paraben were detected in all samples. The highest measured concentration corresponded to methyl paraben, 11.77 ng/g fresh weight. Reported concentrations of benzophenone-4 and benzyl paraben constitute the first evidence about their accumulation in placenta. The results obtained corroborate that foetuses are exposed to a wide diversity of UV filters and PBs via the placenta.     

Comprehensive study of endocrine disrupting compounds using grab and passive sampling at selected wastewater treatment plants in South East Queensland, Australia

Chemical (gas chromatography-mass spectrometry, GC-MS) and biological (E-Screen assay) analyses were used to determine the concentrations of 15 endocrine disrupting compounds (EDCs) and estrogen equivalent (EEq) in grab and passive samples from five municipal wastewater treatment plants (WWTPs) in South East Queensland, Australia. EEq concentrations derived by E-Screen assays for the grab samples were between 108-356 ng/L for the influents and < 1-14.8 ng/L for the effluents with the exception of one effluent sample which was at 67.8 ng/L EEq. The EDC concentrations and EEq values for the passive samples were several times lower than those of the grab samples: a decrease probably caused by, but not limited to biofouling, low flow rate, biodegradation and temperature which can progressively reduce the uptake of compounds into the sampler. At this stage, grab sampling is the most reliable method for field monitoring; nevertheless, passive sampler is a useful sampling tool but the method requires more research to ensure that the information obtained can be interpreted appropriately. Although alkylphenols and phthalates were detected at higher concentrations in the wastewater samples as compared to natural hormones, the environmental risk may be negligible as their estrogenic potencies are several orders of magnitude lower than that of the natural estrogens. In most wastewater samples, the natural estrogens contributed to 60% or more of the EEq value. Removal efficacy of most estrogenic and xenoestrogenic compounds from the conventional activated sludge or biological nutrient removal (BNR) WWTPs monitored in this study was in the range of 80-> 99%. The efficiency of the WWTPs in removing estrogenic activity was > 95%. The EEqs of the E-Screen and those calculated from the results of extensive chemical analyses using the estradiol equivalency factors were comparable for most of the WWTPs samples.     

Occurrence,fate and ecological risk of five typical azole fungicides as therapeutic and personal care products in the environment: A review

Azole fungicides are widely used to treat fungal infection in human. After application, these chemicals may reach to the receiving environment via direct or indirect discharge of wastewaters, thus posing potential risks to non-target organisms. We aimed to review the occurrence, fate and toxicological effects of some representative household azole fungicides in the environment. Azole fungicides were widely detected in surface water and sediment of the aquatic environment due to their incomplete removal in wastewater treatment plants. These chemicals are found resistant to microbial degradation, but can undergo photolysis under UV irradiation. Due to different physiochemical properties, azole fungicides showed different environmental behaviors. The residues of azole fungicides could cause toxic effects on aquatic organisms such as algae and fish. The reported effects include regulation changes in expression of cytochrome P450-related genes and alteration in CYP450-regulated steroidogenesis causing endocrine disruption in fish. Further studies are essential to investigate the removal of azole fungicides by advanced treatment technologies, environmental fate such as natural photolysis, and toxic pathways in aquatic organisms.     

Genotoxic effects of binary mixtures of xenoandrogens (tributyltin, triphenyltin) and a xenoestrogen (ethinylestradiol) in a partial life-cycle test with Zebrafish (Danio rerio)

A partial life-cycle test with the model fish Danio rerio was performed in order to evaluate the genotoxic potential of binary mixtures of xenoandrogenic (tributyltin--TBT; triphenyltin--TPT) and an estrogenic compound (ethinylestradiol--EE2). Five days post-fertilisation larvae were diet-exposed to environmental relevant concentrations of TBT and TPT (25 ng/g-100 ng/g), and water-exposed to ethinylestradiol (3.5 ng/L) for a four-month period; binary mixtures of TBT plus EE2 and TPT plus EE2 were run in parallel. The erythrocytic nuclear abnormalities (ENA) assay in circulating erythrocytes was used to evaluate genotoxicity in the end of the four-month exposure period. A significant increase (p<0.05, Kruskall-Wallis non-parametric ANOVA) in ENA frequency, in comparison with control animals, was observed in those animals exposed to TBT and TPT (the highest doses only), and to EE2 and the binary mixtures, although neither synergistic nor additive effects of the tested compounds were evident. Overall, the results clearly indicate that chronic exposure to low levels of TBT, TPT, EE2 and binary mixtures of TBT plus EE2 and TPT plus EE2 are genotoxic to zebrafish, which may suggest that wild fish populations may be under increased DNA damage in areas contaminated by these endocrine disrupting chemicals.     

Which compounds contribute most to elevated airborne exposure and corresponding health risks in the Western Balkans?

A majority of ongoing monitoring of persistent organic pollutants (POPs) is currently focused on chemicals emphasized in the Stockholm Convention. Quantitative detection of other substances (especially those with numerous anthropogenic sources such as polyaromatic hydrocarbons (PAHs)) is, however, also needed since their concentrations are usually several orders of magnitude higher. A goal of this study was to determine how various groups of compounds contribute to total human health risks at the variety of sampling sites in the region of Western Balkan. Distribution of the risks between the gas and particulate phases was also addressed. Results showed that inhalation exposure to organochlorine pesticides (OCPs) does not represent a significant risk to humans, while polychlorinated biphenyls (PCBs) re-volatilized to the atmosphere from contaminated soils and buildings can pose a problem. PCB evaporation from primary sources (currently used PCB-filled transformers or non-adequate storage facilities) generally resulted in much higher atmospheric concentrations than evaporation from the secondary sources (soils at the sites of war destructions). A majority of the human health risks at the urban sites were associated with PAHs. Between 83 and 94% of the cumulative risk at such sites was assigned to chemicals sorbed to particles, and out of it, PAHs were responsible for 99%.     

Non-invasive matrices in human biomonitoring: a review

Humans and other living organisms are exposed to a variety of chemical pollutants that are released into the environment as a consequence of anthropogenic activities. Environmental pollutants are incorporated into the organism by different routes and can then be stored and distributed in different tissues, which leads to an internal concentration that can induce different alterations, adverse effects and/or diseases. Control measures should be taken to avoid these effects and human biomonitoring is a very useful tool that can contribute to this aim. Human biomonitoring uses different matrices to measure the target chemicals depending on the chemical, the amount of matrix necessary for the analysis and the detection limit (LOD) of the analytical technique. Blood is the ideal matrix for most chemicals due to its contact with the whole organism and its equilibrium with organs and tissues where chemicals are stored. However, it has an important disadvantage of being an invasive matrix. The development of new methodology and modern analytical techniques has allowed the use of other matrices that are less or non-invasive, such as saliva, urine, meconium, nails, hair, and semen or breast milk. The presence of a chemical in these matrices reflects an exposure, but correlations between levels in non-invasive matrices and blood must be established to ensure that these levels are related to the total body burden. The development of new biomarkers that are measurable in these matrices will improve non-invasive biomonitoring. This paper reviews studies that measure Cd, Pb, Hg, polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polycyclic aromatic hydrocarbons (PAHs), polybrominated diphenyl ethers (PBDEs), organochlorine pesticides and phthalates in non-invasive matrices, the most used techniques for measurements and what alternative techniques are available.     

Alteration of steroidogenesis in H295R cells by organic sediment contaminants and relationships to other endocrine disrupting effects

A novel bioassay with the human adrenocortical carcinoma cell line H295R can be used to screen for endocrine disrupting chemicals that affect the expression of genes important in steroidogenesis. This assay was employed to study the effects of organic contaminants associated with the freshwater pond sediments collected in the Ostrava-Karvina region, Czech Republic. The modulation of ten major genes involved in the synthesis of steroid hormones (CYP11A, CYP11B2, CYP17, CYP19, 17betaHSD1, 17betaHSD4, CYP21, 3betaHSD2, HMGR, StAR) after exposure of H295R cells to sediment extracts was investigated using quantitative real-time polymerase chain reaction (PCR). Crude sediment extracts, containing high concentrations of polycyclic aromatic hydrocarbons (PAHs) and moderate amounts of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) significantly stimulated expression of the CYP11B2 gene (up to 10-fold induction), and suppressed expression of 3betaHSD2 and CYP21 genes. A similar pattern was observed with the extracts after treatment with concentrated sulfuric acid to remove labile chemicals (including PAHs) leaving only persistent PCBs, OCPs and potentially PCDD/Fs. Comparison of the results with other mechanistically based bioassays (arylhydrocarbon receptor, AhR, mediated responses in H4IIE-luc cells, and estrogen receptor mediated effects in MVLN cells) revealed significant endocrine disrupting potencies of organic contaminants present in the sediments (most likely antiestrogenicity). Pronounced effects were observed particularly in sediment extracts from the Pilnok Pond which harbors an unusual intersexual population of the narrow-cawed crayfish Pontastacus leptodactylus (Decapoda, Crustacea). This pilot study provided the first experimental evidence of the wider application of the H295R bioassay for screening complex environmental samples, and the results support the hypothesis of chemical-induced endocrine disruption in intersexual crayfish.     

Occurrence and fate of hormone steroids in the environment

Hormone steroids are a group of endocrine disruptors, which are excreted by humans and animals. In this paper, we briefly review the current knowledge on the fate of these steroids in the environment. Natural estrogenic steroids estrone (E1), 17beta-estradiol (E2) and estriol (E3) all have a solubility of approximately 13 mg/l, whereas synthetic steroids 17alpha-ethynylestradiol (EE2) and mestranol (MeEE2) have a solubility of 4.8 and 0.3 mg/l, respectively. These steroids have a moderate binding on sediments and are reported to degrade rapidly in soil and water. Estrogenic steroids have been detected in effluents of sewage treatment plants (STPs) in different countries at concentrations ranging up to 70 ng/l for E1, 64 ng/l for E2, 18 ng/l for E3 and 42 ng/l for EE2. E2 concentrations in river waters from Japan, Germany, Italy and the Netherlands ranged up to 27 ng/l. In addition, E2 concentrations ranging from 6 to 66 ng/l have also been measured in mantled karst aquifers in northwest Arkansas. This contamination of ground water has been associated with poultry litter and cattle manure waste applied on the land. Although hormone steroids have been detected at a number of sources worldwide, currently, there is limited data on the environmental behaviour and fate of these hormone steroids in different environmental media. Consequently, the exposure and risk associated with these chemicals are not adequately understood.