In Search of Mercury Lost from Sediments in a Previously Contaminated Coastal Area, Harboøre Tange, Denmark.

Concentrations of mercury in sediment and benthic invertebrate fauna of Nissum Broad, North-western Jutland, Denmark were investigated. The western coast of Nissum Broad is Harboøre Tange, along which heavy mercury contamination - caused by discharge from production of mercury containing seed dressers in the 1950 and 1960s - was documented in the 1980s. Recent investigations showed marked decreases in mercury contamination in the near shore sediments along Harboøre Tange since the 1980s and the present investigation was initiated to learn if the loss of mercury from Harboøre Tange had led to an increased mercury contamination in the neighbouring marine area, Nissum Broad. Mercury concentrations in the surface sediment correlated with the content of organic matter and the slope of the regression is a good indicator for the degree of mercury contamination. Average mercury concentrations in the upper 5 cm of the sediments ranged between 0.9 and 71 ng g- 1 dry weight (dw) with only 1 station exceeding the Background Assessment Concentration of 70 ng g- 1 dw. Average mercury concentrations in blue mussels Mytilus edulis (169-260 ng g- 1 dw) and periwinkles Littorina littorea (66-203 ng g- 1 dw) exceeded those in uncontaminated areas and the Environmental Quality Standard of approximately 100 ng g- 1 dw. Present sediment mercury concentrations in Nissum Broad are approximately half of what they were in the 1980s, rendering it unlikely that mercury lost from Harboøre Tange has been deposited there. Sediment and organism concentrations did not show any correlation.

Investigation of the in vivo estrogenicity of the UV-filters 4-methylbenzylidene camphor and octyl methoxy cinnamate in rainbow trout (Oncorhynchus mykiss).

The estrogenic activity of the chemical UV-filters, 4-methylbenzylidene camphor (4-MBC) and octyl methoxy cinnamate (OMC) was investigated in an in vivo rainbow trout (Oncorhynchus mykiss) assay. Plasma vitellogenin concentrations were quantified by means of an Enzyme Linked Immuno Sorbent Assay (ELISA) in juvenile rainbow trout before and after intraperitoneal injection of the test compounds. Injection of 4-MBC on day 0, 3, 6 and 10 in the exposure period caused dose and time dependent increases in the concentration of plasma vitellogenin. Significant elevation of vitellogenin concentrations in plasma was demonstrated from 151 mg 4-MBC kg-1 injection-1. Logistic regression analysis relating the percentage of responding fish to the injected dose of 4-MBC resulted in ED10, ED50 and ED90 values of 37, 115 and 194 mg kg-1 injection-1, respectively, after 14 days of exposure (4 injections). Injections with OMC (up to 202 mg kg-1 injection-1) did not result in a statistically significant response in groups of exposed fish, although some individual fish showed elevated concentrations of vitellogenin in plasma. The results confirm that 4-MBC is estrogenic in fish in vivo.

Mercury (Hg2+) interferes with physiological adaptations to freezing in the arctic earthworm Enchytraeus albidus.

Freezing temperatures is an important stressor in the arctic regions and has a significant influence on the population dynamics and geographic distribution of terrestrial invertebrates. Toxic metals in the environment can interfere with protective cold-acclimation responses of organisms. It is therefore important to evaluate the combined effects of cold stress and environmental contaminants. Here, we aimed to investigate the effects of Hg (HgCl2) on various physiological aspects of freeze-tolerance in the earthworm (Enchytraeus albidus). We measured the levels of the cryoprotectant glucose, the glycogen content (source of glucose molecules for cryoprotection and fuel for metabolism), and changes in the composition of membrane phospholipid fatty acids (PLFA) as an indicator of lipid peroxidation. Freezing at -6 °C had no effect on survival in uncontaminated soil, however, survival of freezing in Hg contaminated soil was clearly reduced, especially at extended exposure times. Thus, the LC50 value in frozen soil decreased from 8.3 mg Hg kg-1 (when exposed for 17 days) to only 4.2 mg Hg kg-1 after 36 days' exposure indicating that combined effects of Hg and freezing became larger at prolonged exposure times. Hg caused a depletion of glycogen reserves (almost 50% at 12 mg kg-1 dry soil), but despite this effect worms were able to maintain a constant cryoprotectant level (about 0.12 mg glucose mg-1 dry weight) at all Hg concentrations. Hg had clear negative effects on the proportion of unsaturated PLFAs, which could be an indication of lipid peroxidation. Since a high proportion of unsaturated fatty acids in the membrane is important for invertebrate freeze-tolerance, our results suggest that the negative effect of Hg on freeze-tolerance in E. albidus is related to degraded membrane functionality at low temperature.

Manufacturing doubt about endocrine disrupter science--A rebuttal of industry-sponsored critical comments on the UNEP/WHO report "State of the Science of Endocrine Disrupting Chemicals 2012".

We present a detailed response to the critique of "State of the Science of Endocrine Disrupting Chemicals 2012" (UNEP/WHO, 2013) by financial stakeholders, authored by Lamb et al. (2014). Lamb et al.'s claim that UNEP/WHO (2013) does not provide a balanced perspective on endocrine disruption is based on incomplete and misleading quoting of the report through omission of qualifying statements and inaccurate description of study objectives, results and conclusions. Lamb et al. define extremely narrow standards for synthesizing evidence which are then used to dismiss the UNEP/WHO 2013 report as flawed. We show that Lamb et al. misuse conceptual frameworks for assessing causality, especially the Bradford-Hill criteria, by ignoring the fundamental problems that exist with inferring causality from empirical observations. We conclude that Lamb et al.'s attempt of deconstructing the UNEP/WHO (2013) report is not particularly erudite and that their critique is not intended to be convincing to the scientific community, but to confuse the scientific data. Consequently, it promotes misinterpretation of the UNEP/WHO (2013) report by non-specialists, bureaucrats, politicians and other decision makers not intimately familiar with the topic of endocrine disruption and therefore susceptible to false generalizations of bias and subjectivity.

17ß-estradiol causes abnormal development in embryos of the viviparous eelpout.

Elevated frequencies of malformations among the offspring of Baltic eelpout (Zoarces viviparus) have been observed in aquatic environments receiving high anthropogenic input suggesting that manmade chemicals could be the causative agent. However, causal links between exposure to chemicals and abnormal development have never been confirmed in laboratory experiments. The purpose of this study was to investigate if exposure to 17ß-estradiol (E2) causes abnormal development in larvae of the viviparous eelpout. Wild female eelpout were collected immediately after fertilization and exposed to E2 concentrations ranging from 5.7 to 133 ng L(-1) for 6 weeks in a flow through test system. The experiment shows that E2 concentrations of 53.6 and 133 ng L(-1) cause severe abnormal development among eelpout embryos. Reduced amount of ovarian fluid and increased weight of the ovarian sac indicate disturbance of ovarian function. Female plasma concentrations of E2 and vitellogenin increase in a monotonic concentration-response relationship with significant induction in the low concentration range. Our findings support the plausibility that the abnormal development among eelpout embryos encountered in monitoring programs may actually be caused by exposure to chemicals in the environment.

A path forward in the debate over health impacts of endocrine disrupting chemicals.

Several recent publications reflect debate on the issue of "endocrine disrupting chemicals" (EDCs), indicating that two seemingly mutually exclusive perspectives are being articulated separately and independently. Considering this, a group of scientists with expertise in basic science, medicine and risk assessment reviewed the various aspects of the debate to identify the most significant areas of dispute and to propose a path forward. We identified four areas of debate. The first is about the definitions for terms such as "endocrine disrupting chemical", "adverse effects", and "endocrine system". The second is focused on elements of hormone action including "potency", "endpoints", "timing", "dose" and "thresholds". The third addresses the information needed to establish sufficient evidence of harm. Finally, the fourth focuses on the need to develop and the characteristics of transparent, systematic methods to review the EDC literature. Herein we identify areas of general consensus and propose resolutions for these four areas that would allow the field to move beyond the current and, in our opinion, ineffective debate.

Science and policy on endocrine disrupters must not be mixed: a reply to a "common sense" intervention by toxicology journal editors.

The "common sense" intervention by toxicology journal editors regarding proposed European Union endocrine disrupter regulations ignores scientific evidence and well-established principles of chemical risk assessment. In this commentary, endocrine disrupter experts express their concerns about a recently published, and is in our considered opinion inaccurate and factually incorrect, editorial that has appeared in several journals in toxicology. Some of the shortcomings of the editorial are discussed in detail. We call for a better founded scientific debate which may help to overcome a polarisation of views detrimental to reaching a consensus about scientific foundations for endocrine disrupter regulation in the EU.

Selenium reduces the retention of methyl mercury in the brown shrimp Crangon crangon.

Methyl mercury accumulated at the top of aquatic food chains constitutes a toxicological risk to humans and other top predators. Because the methyl mercury enters the aquatic food chains at the lower trophic levels, uptake and elimination processes at these levels affect the methyl mercury content at the higher levels. Selenium modulates the biokinetics of mercury in aquatic organisms in fairly complex ways, increasing mercury retention in some aquatic mammals, but decreasing methyl mercury retention in fish. However, it is not known if selenium modulates methyl mercury accumulation at lower trophic levels in aquatic food chains. Here, we show that selenium administered via the food augments the elimination of methyl mercury from marine shrimp and that the effect is dose-dependent, demonstrable down to natural selenium concentrations in aquatic food items. Selenite, seleno-cystine, and seleno-methionine exert this effect but selenate does not. Our results suggest that the selenium naturally present at the lower trophic levels in marine food chains may play an essential role as a modifier of methyl mercury accumulation at these levels, thereby potentially also affecting biomagnification of methyl mercury toward the higher trophic levels in the aquatic food chains.

Dietary selenium reduces retention of methyl mercury in freshwater fish.

Adverse effects from organic mercury transported along aquatic food chains are health issues in humans and other top predators. Methyl mercury in organisms at the lower food chain levels is eliminated slowly, and laboratory studies have not clarified the role of selenium in the retention of methyl mercury in fish. Here, we investigated the effects of dietary selenium on the retention of organic and inorganic mercury in freshwater fish. Addition of selenite to the food augmented elimination of methyl mercury (but not inorganic mercury) from goldfish Carassius auratus in a dose dependent manner; selenite caused methyl mercury to be lost from the general body rather than from any specific organ. Seleno-cystine and seleno-methionine (but not selenate) likewise promoted elimination of methyl mercury from goldfish. The threshold for the augmenting effect of selenite on the elimination of methyl mercury in the zebra fish Danio rerio was 0.95 µg Se g(-1) food; higher concentrations reduced retention of methyl mercury in a dose dependent manner. Selenium concentrations in the food approaching natural background levels increase the elimination of methyl mercury from fish. Thus, selenium levels in a given aquatic food chain may affect mercury contamination along the food chain.

Exposure to methylmercury and inorganic mercury in the food does not lead to trophic magnification in the sea star Asterias rubens.

Methylmercury accumulated at the top of aquatic food chains constitutes a toxicological risk to humans and other top predators. Biomagnification of methylmercury takes place among vertebrates at the higher trophic levels, but this process is less elucidated in benthic invertebrates at the lower trophic levels. Therefore, we investigated the accumulation from food and elimination of methylmercury and inorganic mercury in the benthic sea star Asterias rubens (L.) - a representative of trophic level ~3 - in laboratory experiments. Sea stars fed over 49 days with contaminated mussels (Mytilus edulis) accumulate methylmercury and inorganic mercury to the highest concentrations in the digestive glands, the pyloric caeca, less in stomach, gonad, tube feet, aboral body wall and not to detectable levels in the coelomic fluid. Concerning whole body contents, steady states were reached for both methylmercury and inorganic mercury during the 7-week feeding period and the sea stars reached approximately ½ and » of the concentrations in the mussel food for the two mercury forms, respectively. Half-lives for the elimination of the two mercury forms varied between 45 and 173 days in a 140-d elimination period following the feeding period; inorganic mercury was eliminated faster than methylmercury. Examination of total mercury concentrations in field-collected sea stars confirmed this lack of trophic magnification in relation to the major food items, soft parts of molluscs. We suggest that mercury is not trophically magnified in sea stars 1) because they eliminate methylmercury faster than larger fish and decapod crustaceans and 2) maybe more importantly, because inorganic mercury with its faster elimination constitutes a larger fraction of the total mercury in the food at the lower trophic levels - as opposed to methylmercury which dominates at the higher trophic levels.

Effect of size on concentrations and cadmium inducibility of metallothionein in the shore crab Carcinus maenas.

Metallothionein (MT) plays an important role in protecting organisms from the adverse effects of Cd, Hg, Zn and Cu. Investigations on mammals show variations in metallothionein concentrations and inducibility with age. This has never been investigated in invertebrates, and we determined the concentrations and inducibility of metallothionein in gills and midgut gland of different size classes of shore crabs from uncontaminated areas. Metallothionein concentrations in gills and midgut gland ranged between 30 and 40 µg g-1 dry weight with no differences among the different size classes. Concentrations of cadmium, copper and zinc in the midgut gland increased with increasing size of the crabs when the concentrations were expressed on a dry weight basis; water content in the midgut gland increased with the size and only the cadmium concentration increased with size when concentrations were expressed on wet weight basis. There was an inverse relationship between metallothionein and both copper and cadmium concentrations. Smaller crabs exposed to 1 mg Cd L-1 accumulated higher concentrations of cadmium in midgut gland and gills than larger ones and metallothionein concentrations in the midgut gland were higher in the smaller crabs. However, the increase in metallothionein concentration per accumulated unit of cadmium showed a linear increase with the size of the crabs. The ratio [Cd]midgut/[Cd]gills decreased with the size of the crabs. The overall conclusion is that baseline metallothionein concentrations do not change with age in shore crabs, but that the inducibility of metallothionein upon cadmium challenge does.

Mercury levels in humpback whales, and other Southern Ocean marine megafauna.

Mercury is a known potent neurotoxin. The biogeochemical cycle of mercury in the remote Antarctic region is still poorly understood, with Polar climate change contributing added complexity. Longitudinal biomonitoring of mercury accumulation in Antarctic marine megafauna can contribute top-down insight into the bio-physical drivers of wildlife exposure. The bioaccumulative nature of organic mercury renders high trophic predators at the greatest risk of elevated exposure. Humpback whales represent secondary consumers of the Antarctic sea-ice ecosystem and an ideal biomonitoring species for persistent and bioaccumulative compounds due to their extended life-spans. This study provides the first results of mercury accumulation in humpback whales, and places findings within the context of mercury accumulation in both prey, as well as six other species of Antarctic marine megafauna. Combined, these findings contribute new baseline information regarding mercury exposure to Antarctic wildlife, and highlights methodological prerequisites for routine mercury biomonitoring in wildlife via non-lethally biopsied superficial tissues.

Effects of 17beta-trenbolone in male eelpout Zoarces viviparus exposed to ethinylestradiol.

To evaluate the interaction between 17beta-trenbolone (TB) and 17alpha-ethinylestradiol (EE2), male eelpout, Zoarces viviparus, was exposed for 21 days (April to May 2008) to 5 ng l(-1) EE2 and 5 or 20 ng l(-1) TB, separately or in combination in a flow-through SW system. The effects on hepatosomatic (HSI) and gonadosomatic index (GSI), plasma vitellogenin (Vtg) concentration, gonadal histology, hepatic and testicular Vtg mRNA and estrogen receptor (ERalpha) mRNA expression were investigated. No effects on HSI were observed. A significant decrease was observed in the GSI of all males exposed to EE2 (<0.7%) when compared to controls (1.4%). Histological alterations and immature stages were observed in the testis of all exposed males; however, males exposed to EE2 were the most affected. Increased tubule number and proportionally decreased tubule diameter were observed in the testis of all EE2 groups. No effects in Vtg mRNA expression were observed in the testis; however, a significant decrease in testis ERalpha mRNA was observed in males exposed to 20 ng l(-1) TB. The groups exposed to EE2 showed a significant increase in plasma Vtg (>300-fold), hepatic Vtg mRNA (>450-fold), and ERalpha mRNA (>100-fold) when compared to controls. This study shows that lower concentrations of 17beta-trenbolone are unable to counteract the EE2 estrogenic effects when the exposure is simultaneous.

Expression of prostaglandin synthases (pgds and pges) during zebrafish gonadal differentiation.

The present study aimed at elucidating whether the expression pattern of the membrane bound form of prostaglandin E2 synthase (pges) and especially the lipocalin-type prostaglandin D2 synthase (pgds) indicates involvement in gonadal sex differentiation in zebrafish as has previously been found in other species. In mice and chicken, the lipocalin-type Pgds is specifically expressed in pre-Sertoli cells just after Sry and Sox9 and is involved in masculinisation of the developing testis. Furthermore, Pges are implicated in female reproduction including follicular development and ovulation. In this study, a sexually dimorphic expression of pgds was found in gonads of adult zebrafish with expression in testis but not in ovaries. To determine whether the sex-specific expression pattern of pgds was present in gonads of juvenile zebrafish and therefore could be an early marker of sex in zebrafish, we microdissected gonads from four randomly selected individual zebrafish for every second day in the period 2-20 days post hatch (dph) and 0-1 dph. The temporal expression of pgds and pges was investigated in the microdissected gonads, however, no differential expression that could indicate sex-specific difference between individual juvenile zebrafish was observed.

Elevated mercury concentrations in biota despite reduced sediment concentrations in a contaminated coastal area, Harboøre Tange, Denmark.

Metals sequestered in coastal sediments are normally considered to be stable, but this investigation shows - somewhat surprisingly - that mercury concentrations in a previously contaminated area, Harboøre Tange, Denmark, have decreased since the 1980s. Mercury concentrations were determined in sediment and benthic biota and present values were compared to values in the 1980s and values from areas without known; history of mercury contamination. Concentrations in both the upper 20 cm of the sediments and; biota are considerably lower now compared to latest monitoring (1980s). Sediment. concentrations at most locations have decreased from the 100-300 ng Hg g-1 dry weight (dw) level to levels below the Background Concentration (BC) of 50 ng Hg g-1 dw defined by Oslo-Paris Convention for the Protection of the Marine Environment of the North-East Atlantic; some stations are at the 2-10 ng Hg g-1 dw level characteristic of Danish coastal sediments with no known history of mercury contamination. Concentrations of mercury in the benthic biota along Harboøre Tange have also decreased since the 1980s but despite the lowered mercury concentrations in the sediments, concentrations in most samples of benthic invertebrate fauna still exceed those in uncontaminated coastal areas and also the Environmental Quality Standard (EQS) of 20 ng Hg g-1 wet weight (≈100 ng Hg g-1 dry weight) defined by the European Union's Water Framework Directive. Concentration ranges in selected organisms are: (Harboøre Tange l980s/Harboøre Tange now/uncontaminated areas - given in ng Hg g-1 dw): Periwinkles Littorina littorea 9000/150-450/55-77, blue mussels Mytilus edulis up to 9000/300-500/40-170, cockles Cerastoderma edule up to 8000/400-1200/200, brown shrimp Crangon crangon 700-2200/150-450/47, eelgrass Zostera marina up to 330/25-70/12. The present results - together with a literature review - show that a simple and straight forward relationship between the concentrations of mercury in sediment and benthic organisms does not necessarily exist.

Laser capture microdissection of gonads from juvenile zebrafish.

BACKGROUND: Investigating gonadal gene expression is important in attempting to elucidate the molecular mechanism of sex determination and differentiation in the model species zebrafish. However, the small size of juvenile zebrafish and correspondingly their gonads complicates this type of investigation. Furthermore, the lack of a genetic sex marker in juvenile zebrafish prevents pooling gonads from several individuals. The aim of this study was to establish a method to isolate the gonads from individual juvenile zebrafish allowing future investigations of gonadal gene expression during sex determination and differentiation. METHODS: The laser capture microdissection technique enables isolation of specific cells and tissues and thereby removes the noise of gene expression from other cells or tissues in the gene expression profile. A protocol developed for laser microdissection of human gonocytes was adjusted and optimised to isolate juvenile zebrafish gonads. RESULTS: The juvenile zebrafish gonad is not morphologically distinguishable when using dehydrated cryosections on membrane slides and a specific staining method is necessary to identify the gonads. The protocol setup in this study allows staining, identification, isolation and subsequent RNA purification and amplification of gonads from individual juvenile zebrafish thereby enabling gonadal gene expression profiling. CONCLUSION: The study presents a protocol for isolation of individual juvenile zebrafish gonads, which will enable future investigations of gonadal gene expression during the critical period of sex differentiation. Furthermore, the presented staining method is applicable to other species as it is directed towards alkaline phosphatase that is expressed in gonocytes and embryonic stem cells, which is conserved among vertebrate species.

Expression profiles for six zebrafish genes during gonadal sex differentiation.

BACKGROUND: The mechanism of sex determination in zebrafish is largely unknown and neither sex chromosomes nor a sex-determining gene have been identified. This indicates that sex determination in zebrafish is mediated by genetic signals from autosomal genes. The aim of this study was to determine the precise timing of expression of six genes previously suggested to be associated with sex differentiation in zebrafish. The current study investigates the expression of all six genes in the same individual fish with extensive sampling dates during sex determination and -differentiation. RESULTS: In the present study, we have used quantitative real-time PCR to investigate the expression of ar, sox9a, dmrt1, fig alpha, cyp19a1a and cyp19a1b during the expected sex determination and gonadal sex differentiation period. The expression of the genes expected to be high in males (ar, sox9a and dmrt1a) and high in females (fig alpha and cyp19a1a) was segregated in two groups with more than 10 times difference in expression levels. All of the investigated genes showed peaks in expression levels during the time of sex determination and gonadal sex differentiation. Expression of all genes was investigated on cDNA from the same fish allowing comparison of the high and low expressers of genes that are expected to be highest expressed in either males or females. There were 78% high or low expressers of all three "male" genes (ar, sox9a and dmrt1) in the investigated period and 81% were high or low expressers of both "female" genes (fig alpha and cyp19a1a). When comparing all five genes with expected sex related expression 56% show expression expected for either male or female. Furthermore, the expression of all genes was investigated in different tissue of adult male and female zebrafish. CONCLUSION: In zebrafish, the first significant peak in gene expression during the investigated period (2-40 dph) was dmrt1 at 10 dph which indicates involvement of this gene in the early gonadal sex differentiation of males.

Vitellogenin as a biomarker for estrogenic effects in brown trout, Salmo trutta: laboratory and field investigations.

The sensitivity of juvenile brown trout towards estrogenic chemicals (17beta-estradiol [E2], estrone [E1], 17alpha-ethinylestradiol [EE2], 4-tert-octylphenol [OP], and n-butylparaben [BP]) was tested in laboratory experiments with plasma and liver vitellogenin concentrations as endpoints. Vitellogenin concentrations were also assessed in juvenile brown trout collected in streams affected by agricultural runoff and discharges from scattered houses in the open land. In the laboratory, juvenile brown trout were exposed to the chemicals in flow-through tanks for 7 to 12 d and concentration-response relationships for the induction of vitellogenin synthesis were obtained. The actual exposure concentrations were determined by liquid chromatography-mass spectrometry. The median plasma vitellogenin concentration in first year control brown trout reared in recirculated groundwater was 165 ng/ml with 783 ng/ml as the highest value. The median effective concentration (EC50) values for vitellogenin induction (based on plasma concentrations) were 3.7 ng EE2/L, 15 ng E2/L, 88 ng E1/L, 68 microg BP/L, and 7 microg OP/L. Median effective concentrations derived from liver vitellogenin concentrations were similar. The 166 brown trout caught in the field were mainly first and second year fish and a few third year fish. Plasma vitellogenin concentrations were below 1000 ng/L in 146 of the fish, between 1000 ng/L and 4234 ng/L in 19 fish and 5.3 x 10(6) ng/L in one male fish. Vitellogenin concentrations did not differ between first and second year fish, but were elevated in third year fish. The data may indicate that juvenile (<2 years) trout with plasma vitellogenin concentrations above 1000 ng/ml have had their vitellogenin synthesis induced by exposure to estrogens in the environment. Plasma and liver vitellogenin concentrations were closely correlated in brown trout with elevated vitellogenin concentrations. It is noteworthy, however, that exposure to synthetic estrogens (EE2, BP, and OP) resulted in higher liver concentrations (for the same plasma concentration) than exposure to the natural estrogens E1 and E2.

Vitellogenin concentrations in feral Danish brown trout have decreased: An effect of improved sewage treatment in rural areas?

Feminization of male and juvenile fish because of exposure to estrogens or estrogenic chemicals in effluents from central wastewater treatment plants (WWTPs) is a worldwide issue of concern. Intersex and induction of the female yolk protein, vitellogenin, in male and juvenile fish are robust biomarkers for estrogenic exposure, and feminized fish have been observed downstream of WWTP outlets in many countries. Danish central WWTPs reduce effluent estrogenicity effectively by advanced sewage treatment, and feminizations have not been observed downstream of central WWTP outlets. However, between 2000 and 2004, investigations of Danish streams not receiving sewage from central WWTPs revealed a high variation in vitellogenin concentrations of male juvenile brown trout (Salmo trutta); some individuals had high concentrations, probably as a result of estrogenic point sources, and the plasma concentration was >50 ng mL-1 in 79% of the juvenile males. The streams were reinvestigated in 2010 to 2016, and the average male level had decreased to a hitherto unseen baseline level; in 2010 only 0.7% (one individual) of the males had a vitellogenin concentration >50 ng mL-1 , which could indicate that the estrogenicity of the streams decreased after 2004. We examined possible estrogenic sources in streams unaffected by central WWTP effluents, and found that the reduced vitellogenin levels are most likely explained by a national effort to improve on-site wastewater treatment in scattered houses not connected to central WWTPs. Environ Toxicol Chem 2018;37:839-845. © 2017 SETAC.

Two common mild analgesics have no effect on general endocrine mediated endpoints in zebrafish (Danio rerio).

Mild analgesics such as acetylsalicylic acid (ASA) and acetaminophen (APAP) exert their pain-relieving effect in humans by inhibition of prostaglandin synthesis. Prostaglandins play key roles in developmental and reproductive processes in vertebrates, and in recent years, it has been suggested that weak analgesics might also act as endocrine disrupters. In a set of experiments we investigated if ASA and APAP affect well-established endocrine endpoints in zebrafish (Danio rerio), which is a commonly used model organism in the investigation of endocrine disrupting chemicals. Zebrafish were exposed to APAP (0.22, 2.3, and 30mgL-1) or ASA (0.2, 0.5, 1.7, and 8.2mgL-1) from hatch to sexual maturity in a test design resembling the OECD Fish Sexual Development Test. No effects on sex ratio and vitellogenin levels were observed. Adult zebrafish were exposed to high concentrations (mgL-1) of ASA or APAP for eight or 14days. ASA reduced the levels of prostaglandin E2, but had no effect on the concentration of 11-ketotestosterone and vitellogenin. Overall, ASA decrease prostaglandin E2 concentrations, but well-established endpoints for endocrine disruption in zebrafish are generally not affected by aquatic exposure neither during development nor adulthood. According to the WHO/IPCS definition of an endocrine disrupter, the present results do not define APAP and ASA as endocrine disrupters.