Effect of water pH and salinity on the survival of eggs and larvae of the euryhaline teleost, Gasterosteus aculeatus L

The effect of water acidification and salinity on the survival of the eggs and larvae of the euryhaline teleost, Gasterosteus aculeatus, was studied using tap water that was low in calcium and high in total aluminium as the basic medium. The survival of eggs from fertilisation to hatching increased with an increase in pH from 4.5 to 6.5. Survival was higher at a given pH at higher salinities (equivalent to 5% and 10% seawater) than in tap water. The salinity experienced by the female prior to spawning also had an effect on egg survival. Survival for 7 days after hatching was high except at the lowest pH levels. The salinity experienced by the female had no significant effect on larval survival.     

Influence of an insect growth regulator on the larval development of an estuarine shrimp

The influence of methoprene, an insect growth regulator used in mosquito control, on larval development of the estuarine grass shrimp (Palaemonetes pugio) was examined in the laboratory. No grass shrimp larvae successfully completed metamorphosis when continuously exposed to 1000 microg methoprene litre(-1). Completion of larval metamorphosis was significantly reduced by exposure to 100 microg litre(-1) of the isomeric mixture (R,S)-methoprene but not the single isomer formulation (S)-methoprene. No statistically significant difference was revealed, however, in ability to inhibit metamorphosis between these two isomeric types across the broad range of exposure concentrations from 0.1 to 1000.0 microg litre(-1). The first two larval stages and the final premetamorphic larval stage were more sensitive to methoprene toxicity than intermediate larval stages. Methoprene exposure did not alter either the duration of total larval development or the total number of larval stages prior to metamorphosis.     

The influence of plantation forestry on the pH and aluminium concentration of upland welsh streams: a re-examination

Previous studies relating forest presence to stream acidity and aluminium concentration were based on small numbers of catchments, often precluding the elimination of confounding influences on stream chemistry, such as geology or soil type. Spatial patterns in aluminium and pH data from 113 Welsh catchments of contrasting land use were therefore analysed in three different ranges of acid sensitivity (< 10, 10-15, 15-25 mg CaCO(3) litre(-1) total hardness). In each range, pH declined and aluminium increased significantly with increasing percentage forest cover. There was no evidence that the relationships reflected a spurious effect of forest location. Where aluminium concentrations were elevated under forest in a sub-set of 13 streams, aluminium was present predominantly in the labile form, most toxic to fish. Regressions of pH and aluminium on percentage forest cover provide a useful method of assessing the amount of forest in Welsh catchments which might give rise to given chemical conditions (e.g. pH <6, Al > 80 microg litre(-1)), though some difficulties are likely in accurately specifying the conditions desirable for fish or other biota.     

The influence of an insect growth regulator on the larval development of the mud crab Rhithropanopeus harrisii

Effects of the juvenile hormone analogue (S)-methoprene on the larval development and survival of the estuarine mud crab Rhitrhopanopeus harrisii were examined in the laboratory. Crab leave continuously exposed to 1000 microg (S)-methoprene litre(-1) did not survive beyond zoeal stage I. With continuous exposure to 100 microg (S)-methoprene litre(-1), mortality significantly increased through all larval stages except for zoeal stage II, and developmental duration significantly increased through all zoeal stages but not the premetamorphic megalopa. No supernumerary larval stages occurred with exposure (S)-methoprene. Rhithropanopeus harrisii larvae appear to be more sensitive to the single isomer formulation, (S)-methophrene, than to the double isomer formulation, (S)-methoprene, used in previously documented studies.     

Acute and chronic toxicity of triphenyltin hydroxide to fathead minnows (Pimephales promelas) following brief or continuous exposure

Fathead minnow larvae (Pimephales promelas) were exposed to triphenyltin hydroxide (TPTH) during brief (single pulse) or continuous exposure in 96-h and 30-day toxicity tests. The continuous exposure 96-h LC(50) value was 7.1 microg litre(-1). Brief exposures for 12 to 72-h gave 96-h LC(50) values that ranged from 61.8 to 6.0 microg litre(-1), respectively. The continuous exposure 30-day chronic effect concentration, based upon reduced growth, was 0.23 microg litre. Survival was significantly reduced at 2.0 microg litre(-1). Brief exposures for 24, 48, and 72-h in 30-day tests significantly reduced survival and growth at 13.0, 13.0 and 60.0 microg litre(-1) respectively. It is suggested that both toxicant concentration and exposure duration are important factors to consider in the risk assessment of potential pesticide hazards in the environment.     

The mixing zone between limed and acidic river waters: complex aluminium chemistry and extreme toxicity for salmonids

When liming running waters, dosers must compensate for different flow and water qualities and for the downstream inflow from acid tributaries which creates mixing zones. At a certain point in the mixing zone, a constant or fluctuating chemical disequilibrium will appear due to transformation processes. In laboratory assays, over-saturated solutions of aluminium with ongoing active precipitation of aluminium have been found to be especially toxic to fish. Recent experiments in a mixing zone in the limed River Audna, Norway, have confirmed this phenomenon. Atlantic salmon (Salmo salar L.) and sea trout (Salmo trutta L.) smolts were exposed to acid and limed waters and mixtures of the two waters downstream from the point of connection. In the acid tributary (mean values: pH=4.8, Ca=1.3 mg litre (-1)), Ali 236 microg litre(-1)=), LT5) was 22 and 40 h for Atlantic salmon and sea trout, respectively. In the mixing zone (pH=4.8-6.5, Ca=1.2-3.2 mg litre(-1), Ali=50-240 microg litre(-1)), LT50 was 7 h for both species, masking the normal species difference in tolerance. Osmoregulatory failure and rapid gill lesions occurred in the mixing zone as an effect of the transformation of Al into high molecular weight precipitating species. This is the first documentation of the existence of such highly toxic mixing zones in nature, and the results clearly show that the mixing zone is even more toxic to fish than acid aluminium-rich waters.     

Chronic effects of Cd on reproduction of Polypedilum nubifer (Chironomidae) through water and food

Chronic effects of Cd on reproduction of Polypedilum nubifer (Chironomidae) were investigated by Cd-exposure from the egg stage using a flow-through aquarium. No significant effect of Cd on reproduction was observed in the midge larvae which had been exposed to 10 or 20 microg Cd litre(-1). Emergence of the Cd-exposed larvae peaked several days before that of the control, although growth was impaired in 1st and/or 2nd instars. The percentage emergence success decreased to about 46% of the control at 40 microg Cd litre(-1). However, other reproductive processes (adult sex ratio, oviposition success and egg hatchability) were not impaired. The emergence success decreased to less than 3% of the control at 80 microg Cd litre(-1). The emergence success of midge larvae which had been fed Cd-contaminated food with 220 or 1800 microg Cd g(-1) decreased to nearly 60% of the control. However, other reproductive processes were not impaired. Emergence success of larvae exposed to food containing 22 microg Cd g(-1) was unaffected.     

Chemical and biological effects of acid, aluminium and lime additions to a Welsh Hill-stream

Chemical and biological responses to simultaneous additions of acid, aluminium and lime were investigated in contiguous 250m-reaches of a chronically acidic stream in Wales. Treatments were applied for 24 h, and from the upstream end were as follows: zone A-untreated, pH 5.0, 0.37 mg litre(-1) filterable Al; zone B-acidified to pH 4.5, 0.40 mg Al litre(-1) (47% of Al attributed to release from the stream bed due to acid additions); zone C-acidified to pH 4.5 and Al dosed to 0.67 mg litre(-1); zone D-dosed with limestone slurry, resulting in pH 7.2, 0.13 mg Al litre(-1). In all reaches, the chemistry of the interstitial water at depths of 0.15 and 0.3 m never fell below pH 5.5, with corresponding decreases in Al and increases in base cation concentrations. Brown trout, Salmo trutta, and crayfish, Austropotamobius pallipes, held in the stream showed decreases in plasma [Na(+)] and haemolymph [Na(+)], respectively, in all acidic zones (A, B, C): these responses were mitigated by liming (zone D). Thus both chronic and simulated episodic levels of pH and dissolved Al were sub-lethally toxic to test species of aquatic fauna. This experiment also demonstrates a stream bed source/sink of Al, and the availability of a possible refuge from acidic surface waters within the substratum.     

Abundance and population structure of perch (Perca fluviatilis L.) in some acidic Norwegian lakes

Abundance (catch per unit effort, n=3752) and population structure of perch Perca fluviatilis were studied in 30 acidic Norwegian lakes with relation to pH (4.3-5.9), calcium (0.41-2.44 mg litre(-1)), labile aluminium (24-255 microg litre(-1)) and total organic carbon (TOC, 1.7-13.8 mg C litre(-1)). Standard series of bottom gill-nets were used to sample fish populations. A multiple regression analysis showed that abundance was significantly correlated to TOC (P<0.05) and, to a smaller extent, to the Ca in lakes with concentrations between 0.41 and 1.70 mg litre(-1) (P=0.07). The lakes which were inhabited by pike were excluded from the analysis. Recruitment failure seems to be the main cause of reductions in perch numbers in the lakes studied. However, high mortality among adult individuals was also evident, and an episode of fish kill was observed in one of the lakes.     

An assessment of the chronic toxicity of the synthetic pyrethroid, fenvalerate, to Daphnia galeata mendotae, using life tables

The effects of chronic exposure to fenvalerate, a synthetic pyrethroid insecticide, on the cladoceran Daphnia galeata mendotae were evaluated under laboratory conditions. The daphnids were exposed for their entire life cycle to concentrations ranging from 0.005 to 0.1 microg fenvalerate litre(-1). The parameters used to determined toxicity were survival, time to first reproduction, reproductive frequency, number of young per brood, cumulative brood size, intrinsic rate of natural increase (r(m)), generation time (T) and net reproductive rate (R(0)). A concentration of 0.005 microg fenvalerate litre(-1) resulted in an increase in longevity but a decrease in production of young. Higher concentrations caused a decrease in survival. Studies of shorter duration than the complete life cycle of the organisms would not have detected toxicity at such low levels. The intrinsic rate of natural increase, r(m), was not affected by fenvalerate until concentration reached 0.05 microg litre(-1) and r(m) decreased to 0.113 from 0.238. A concentration of 0.01 microg litre(-1) caused the net reproductive rate, R(0), and the generation time, T, to decrease to 73.2 offspring per female and 17.3 days from 125.9 offspring per female and 20.3 days, respectively.     

Assessing chronic toxicity of bisphenol A to larvae of the African clawed frog (Xenopus laevis) in a flow-through exposure system

A number of currently used industrial chemicals are estrogenic, and therefore have potential to disrupt sexual differentiation in vertebrate wildlife during critical developmental windows. We assessed the effect of larval exposure to bisphenol A (BPA) on growth, development and sexual differentiation of the gonad in the African Clawed frog, Xenopus laevis. Larvae were maintained in flow-through conditions at 22 +/- 1 degrees C and exposed to BPA at mean measured concentrations of 0.83, 2.1, 9.5, 23.8, 100, and 497 microg/l, from developmental stages 43/45-66 (completion of metamorphosis). Each test concentration, plus dilution water control (DWC) and positive control (17beta-estradiol (E2), 2.7 microg/l) employed four replicate test vessels with 40 larvae per tank. Individual froglets were removed from test vessels upon reaching stage 66, and the study was terminated at 90 days. Froglets were dissected and sex was determined by inspection of gross gonadal morphology. Test concentrations of BPA had no effect on survival, growth, developmental stage distributions at exposure days 32 and 62, or mean time to completion of metamorphosis, compared to DWC. Analysis of post-metamorphic sex ratio, determined by gross gonadal morphology, indicated no significant deviations from expected (50:50) sex ratio, in DWC or any BPA test concentration. In contrast, exposure of larvae to (E2) resulted in feminisation, with sex ratio deviating significantly (31% male, replicates pooled). Exposure to BPA in the concentration range 0.83-497 microg/l in flow-through conditions had no observable effect on larval growth, development or sexual differentiation (as determined by gross gonadal morphology) in this study.     

Survival of spotted salamander eggs in temporary woodland ponds of coastal Maryland

Temporary ponds on the Atlantic Coastal Plain in maryland were characterized according to water chemistry, rain input, phytoplankton, zooplankton and use by the spotted salamander Ambystoma maculatum during March-October 1983-1984. Neither the number of egg masses per unit of pond surface (abundance) nor the survival of spotted salamander embryos was significantly correlated (P>0.05) with pond pH. Rainfall during May-July significantly increased the hydrogen ion concentration of 5 of 11 ponds evaluated for the impact of rainfall during the previous 48h and the previous week. Survival of egg masses transferred among eight ponds with pH3.66-4.45 and one pond with pH5.18 was significantly reduced (P相似文献   

Toxic Effects of Crude Oil Combined with Oil Cleaner Simple Green on Yolk-sac Larvae and Adult Rainbow Trout Oncorhynchus mykiss (4 pp)

Background, Goal and Scope Cleaner CRYSTAL Simple Green (SG) was used for the cleanup of the oil spill in the Baltic Sea near Lithuania in 2001. No scientific data are available on the effects and consequences of its application for local aquatic life. The aim of this study was to determine and compare sublethal effects of a) solution SG; b) crude oil alone; c) SG in combination with oil on rainbow trout Oncorhynchus mykiss at different stages of its development in laboratory conditions.Methods Laboratory studies were performed on adult rainbow trout (4-day duration) and on yolk-sac larvae (25-day duration) evaluating their biological parameters. Concentrations of water-soluble and thin-dispersed fractions of petroleum hydrocarbons were measured using gas chromatography.Results and Discussion SG solution (0.5 mg/l) did not affect the survival of larvae and adult fish, and no significant changes were determined in respiratory parameters of the exposed larvae and adult fish. The most expressed alterations were found in morphological parameters (a decrease in the average body mass) of larvae and in haematological indices (a decrease in the leukocyte count) of adult fish at the end of the tests. Crude oil (1610 mg/l) did not affect the survival of adult fish during the 4-day exposure. An increase in larvae mortality rate (~ 36%) was recorded at the end of the tests. A significant decrease in the average body mass and heart rate of larvae as well as in gill ventilation frequency of larvae and adult fish were determined. SG combined with oil induced an increase in larval mortality &#61566; 46% of individuals died at the end of the tests. No mortality was recorded in adult fish. The average body mass and heart rate of larvae were significantly decreased. Marked changes were also found in respiratory parameters (gill ventilation frequency of larvae and adult fish significantly decreased, while “coughing” rate increased). A 1-day, 2-day exposure of fish to SG combined with oil induced a significant decrease in the leukocyte count of adult fish, which was also determined at the end of the tests. The augmentation of adverse impact could be explained by the data obtained from our studies. When SG was added into dilution water with crude oil the concentration of petroleum hydrocarbons in the mixture increased 3 ~ 4.5 times after 24 h and 96 h, respectively. Conclusions The comparative study of the effects of crude oil alone, SG and SG combined with oil showed that their toxic effects on fish differed. Oil combined with SG was found to be more toxic to fish (larvae and adults) than SG alone and oil alone. Fish at early stages of development (yolk-sac larvae) were more sensitive to the effects of the compounds studied than adults.Recommendations and Outlook To diminish the negative impact of oil spill cleanup using chemicals on aquatic ecosystems, it is recommended to carry out more comprehensive studies of their effects and after-effects in laboratory conditions using a wide scale of local aquatic organisms. The selected species of the most sensitive aquatic organisms should include those which are unable to escape the impact of combined action of oil and cleaners. Special attention should be directed to the research of effects of these pollutants on studied organisms at their most sensitive stages of life (reproduction, hatching, early stages of development), as after-effects of exposure to pollutants may be observed in future generations.     

Effects of fenoxycarb exposure on complete larval development of the xanthid crab,Rhithropanopeus harrisii

Pest control agents, such as juvenile hormone analogues (JHA), have been developed to limit effects on non-target organisms that co-inhabit insect pest habitats. Rhithropanopeus harrisii, an estuarine xanthid crab, was used to observe the impacts of the JHA, fenoxycarb, on the pattern of complete larval development as well as survival of larvae and successful metamorphosis to first crab stage. Significant mortality occurred in the first of four zoeal stages (after 2-3 days of exposure) at the highest treatment of 240 microg fenoxycarb/l and in megalopae exposed to 48 microg fenoxycarb/l. The time required to metamorphose to the first crab stage was significantly increased for megalopae in all treatments 48 microg/l. This delay in development was sufficient to significantly prolong the entire developmental period from zoea to crabs. Unexposed larvae developed to crabs in an average of 16 days; larvae exposed to >/=48 microg/l required 19-20 days. Reduced survival and extended duration of developing larval stages in the life history of a benthic invertebrate may alter the population dynamics of these organisms in the estuary.     

Bioaccumulation of cadmium by the freshwater isopod Asellus aquaticus (L.) from aqueous and dietary sources

Experiments were conducted to determine the kinetics and relative importance of aqueous and dietary uptake of cadmium by the freshwater isopod Asellus aquaticus (L.). Test animals were exposed during 30 days to aqueous Cd in a continuous flow system (exposure levels: 0.2 - 10 microg litre(-1)) and kept on a diet of previously contaminated Elodea sp. (range of Cd concentrations: 2-350 microg g(-1), dry weight). Preceding semi-static experiments on dosage-control of the dietary factor revealed a rapid uptake of Cd by Elodea, with relatively high concentration factors (CF), which ranged from 4.8 to 5.5 (dry weight log (CF) after 16 days). For Asellus uptake from water appeared to be the predominant route. Highly significant bioconcentration of cadmium from water was observed in the animals, even at exposure levels below 1.0 microg litre(-1). In the various treatments, direct uptake from water accounted for 50-98% of the body burdens after 30 days exposure. The experimental results were described with a first order one-compartment bioaccumulation model. Model parameter estimates (mean +/- standard error) were obtained for rate constant of uptake (560 +/- 110 day(-1)), rate constant of elimination (0.032 +/- 0.017 day(-1)) and assimilation efficiency of Cd uptake from food (1.1 +/- 0.7%). The (dry weight) bioconcentration factor (BCF) and bioaccumulation factor (BAF) for extrapolated steady state conditions were estimated at 18 000 (BCF) and 0.08 (BAF). Experiments conducted at two different pH levels (5.9 versus 7.6) revealed no significant effects of pH on the uptake of aqueous Cd by the isopods. The results are discussed in relation to their potential significance to the field situation.     

Organochlorine pesticides and heavy metals in muscle and ovaries of Gulf coast striped bass (Morone saxatilis) from the Apalachicola River, Florida, USA

Eight female Gulf coast striped bass (Morone saxatilis) broodfish collected for induced spawning from the Apalachicola River below the Jim Woodruff Lock and Dam were analyzed for organochlorine pesticides (OCs) and metals in muscle and ovarian tissues. Chemical analyses revealed that muscle and ovaries contained detectable amounts of OCs and metals. Concentrations of p,p'-DDE, a derivative of the pesticide DDT, in muscle and ovary (0.54 and 0.65 microg/g, respectively) were significantly higher than alpha-chlordane, dieldrin, and p,p'-DDD. The presence of p,p'-DDE, an antiandrogenic compound, in females suggests that the compound also may be present in male striped bass. Concentrations of Cr, Hg, Mg, and Mn were higher in muscle than in ovarian tissues. Concentrations of Hg have almost doubled in muscle tissues (0.85 microg/g) and tripled in ovaries (0. 15 microg/g) in our samples, compared with the data from 1986 to 1989. Organochlorine pesticides and metal contaminants were present in muscle and ovarian tissues of adult females and may have been retarding development of eggs leading to low hatching rates.     

Effects of salinity,pH and temperature on the re-establishment of bioluminescence and copper or SDS toxicity in the marine dinoflagellate Pyrocystis lunula using bioluminescence as an endpoint

Pyrocystis lunula is a unicellular, marine, photoautotrophic, bioluminescent dinoflagellate. This organism is used in the Lumitox bioassay with inhibition of bioluminescence re-establishment as the endpoint. Experiments determined if acute changes in pH, salinity, or temperature had an effect on the organisms' ability to re-establish bioluminescence, or on the bioassay's potential to detect sodium dodecyl sulfate (SDS) and copper toxicity. The re-establishment of bioluminescence itself was not very sensitive to changes in pH within the pH 6-10 range, though reducing pH from 8 to levels below 6 decreased this capacity. Increasing the pH had little effect on Cu or SDS toxicity, but decreasing the pH below 7 virtually eliminated the toxicity of either compound in the bioassay. Lowering the salinity from 33 to 27 per thousandth or less resulted in a substantial decrease in re-establishment of bioluminescence, while increasing the salinity to 43 or 48 per thousandth resulted in a small decline. Salinity had little influence on the bioassay's quantification of Cu toxicity, while the data showed a weak negative relationship between SDS toxicity and salinity. Re-establishment of bioluminescence showed a direct dependence on temperature, but only at 10 degrees C did temperature have an obvious effect on the toxicity of Cu in this bioassay.     

Reproductive status, blood chemistry, gill histology and growth of perch (Perca fluviatilis) in three acidic lakes

Perch (Perca fluviatilis L.) were sampled soon after spawning in three small acidic lakes (pH 4.3-6.1, Al(lab) 5-106 microg litre(-1), Ca2+ 0.01-0.08 mmol litre(-1)) and in one circumneutral lake (pH 5.9-6.4, Al(lab) 4-12 microg litre(-1), Ca2+ 0.06-0.07 mmol litre(-1)) in southern Finland. Due to the delayed spawning of perch in the acidic lakes, sampling in those lakes was performed later than in the reference lake. In spite of that, the gonadosomatic index (GSI) of males in all the acidic lakes was significantly greater than in the reference lake. Of the two lakes with similar low water pH, the effects on reproduction were more prominent in the lake with higher water Al content. The plasma Ca2+ concentrations of females in the acidic lakes were significantly smaller than in the females of the reference lake. The low female:male plasma Ca2+ ratio (1.0-1.32) depicted delay of spawning. Stress in perch in acidic water was also seen in elevated blood haematocrit values, especially in females. On the other hand, a low plasma Cl- level, a common response to acidic water in salmonids, was not detected in perch in the most acidic lakes. The amount of Al accumulated in the gill epithelium was highest in the most acidified lake with high Al concentration, but was also pronounced in a lake with low pH and low Al concentration.     

Effects of long- and short-term exposure to carbaryl on survival, growth and reproduction of Daphnia ambigua

In laboratory experiments a cladoceran, Daphnia ambigua, was exposed for six instars to the insecticide carbaryl at five different concentrations (1, 2, 3, 4, 5 microg litre(-1)) or received short-term (10-h) exposure to 5 microg carbaryl litre(-1) at different life stages of the animal. In the long-term exposure experiments carbaryl at 1 microg litre(-1) did not have any detectable effects on the animals, whereas 2 microg litre(-1) reduced the survival, growth and reproduction of the animals, and higher concentrations killed all the animals examined before they grew to the fifth instar. These results indicate that the range of sublethal concentrations of carbaryl for D. ambigua is narrow. In the short-term exposure experiments the animals did not suffer any damage when they were exposed to the chemical during the egg stage. However, their growth and egg production were reduced when the short-term exposure was conducted during the life stages from the final embryo stage to the third instar. The effect was most marked when the animals were treated during the first instar, indicating that this is the stage when the animals are most sensitive to the chemical.     

Integral assessment of estrogenic potentials of sediment-associated samples

GOAL, SCOPE AND BACKGROUND: Exogenic endocrine-active substances are also called 'Endocrine Disrupting Chemicals' (EDC). They imitate or hinder the function of natural endogenic hormones or disturb the synthesis or the metabolism of hormones or of hormone receptors. The Enzyme-Linked Receptor Assay (ELRA) can detect estrogenic and anti-estrogenic effects at the level of receptor binding and is a useful tool for the integrative detection of contaminant effects. Although the test system has been used repeatedly in sediment assessments, the questions have remained concerning how it responds to variations in the physico-chemical matrix. For some bioassays, the salinity of the sample is a critical factor. This is especially relevant when testing wastewater samples or when sediment-associated samples in the tidal reaches of rivers are tested. Sediments in the tidal reaches of rivers change their salinity several times a day. Against this background, it would be beneficial to have a test procedure of known salinity tolerance. On account of this, the salinity tolerance of the ELRA was tested, assessed with reference substances at several salinity levels, and compared with the E-Screen method and a Yeast Estrogen Screen (YES), which are also frequently applied in environmental testing. The aim of this paper was to explore when the salinity limits within these test procedures are applicable. The trials should reveal the working range to be expected, characterize the salinity-dependent variations in sensitivity of the test, and provide options for methodological adjustments to improve the stability against increased salinity. METHODS: The ELRA was carried out with the human Estrogen Receptor alpha. (ER) using the same principle like a competitive immunoassay based on ligand-protein interaction. However, an essential difference is the use of a physiologically relevant receptor instead of an antibody as a linking protein. The ELRA measures the competition of sample estrogens and anti-estrogens against estradiol supplied as a BSA-coating conjugate for the binding site of dissolved ER. Estradiol or xeno-estrogen binding is quantified by a biotynilated anti-ER antibody and the subsequent measurement of peroxidase activity by a streptavidin-POD-biotin complex. The E-Screen was performed with the human breast cancer cell line MCF-7, which expresses the estrogen receptor constitutively. Cell proliferation depends on binding of estrogens or xeno-estrogens with the receptor. After incubation, estrogen-dependent cell growth was measured by sulforhodamin B staining. The YES was performed with a recombinant yeast strain, transfected with a receptor and a reporter plasmid bearing the estrogen receptor and a vitellogenin gene fused with the reporter gene lacZ. Estrogen or xeno-estrogen-dependent gene induction was measured indirectly by LacZ activity. The salinity levels were simulated in varying concentrations with NaCl from 0 to 40 per thousand or Artificial Sea Water (ASW) from 0 to 32 per thousand. RESULTS: The study characterized the factor 'salinity' for the prospective application fields of the ELRA. With reference substances such as 17-beta-estradiol, the ELRA showed classical sigmoidal concentration-effect relations in a range from 0.05 to 100 microg/l under physiological conditions. After a methodological adjustment to compensate decreasing receptor-binding affinity of estrogens and xeno-estrogens at higher salinity levels, the ELRA became applicable under salinity conditions up to concentrations of 20.5 per thousand. In tests, the ELRA reached under the influence of salinity a mean limit of detection of 0.062 microg/l 17-beta-estradiol. The mean relative inter-test error was around 11%. Above concentrations of 20.5 per thousand there is a risk of false negative assessment. Compared with the E-Screen method using the MCF7 cell line and the yeast estrogen test system (YES), the ELRA shows a lower sensitivity to 17-beta-estradiol. In the E-Screen, the cell proliferation was strongly reduced by sodium chloride induced cytotoxicity. In comparison with the E-Screen, the salinity tolerance of the YES and YAS methods is significantly higher. DISCUSSION: Despite adaption, total salinity tolerance could not be achieved with the ELRA. Freshwater samples were generally appraisable. Higher salinity levels above 20.5 per thousand would tend towards false negative results. The low inter-test error of 11% makes the ELRA suitable for the detection of estrogenic and anti-estrogenic potentials of single substances, substance mixtures, and of environmental samples. CONCLUSIONS: The ELRA is very fast and reproducible, it can be used for high-throughput screening in a microplate format at low cost, it is robust to microbial contamination, and is less susceptible to cytotoxic interferences than cell culture methods. RECOMMENDATIONS AND PERSPECTIVES: In their established form, the YES and the E-Screen methods are not applicable for liquid phase testing at higher salinity conditions. The salinity-adapted test version of the ELRA described here shows a broader working range for samples. Native water samples of more or less brackish origin or high-salinity effluent samples are testable. Results of tests with sediment associated samples of different salinity will be subject of a forthcoming publication.