A comparison of statistical methods for deriving freshwater quality criteria for the protection of aquatic organisms

Species sensitivity distributions (SSDs) are increasingly used in both ecological risk assessment and derivation of water quality criteria. However, there has been debate about the choice of an appropriate approach for derivation of water quality criteria based on SSDs because the various methods can generate different values. The objective of this study was to compare the differences among various methods. Data sets of acute toxicities of 12 substances to aquatic organisms, representing a range of classes with different modes of action, were studied. Nine typical statistical approaches, including parametric and nonparametric methods, were used to construct SSDs for 12 chemicals. Water quality criteria, expressed as hazardous concentration for 5 % of species (HC₅), were derived by use of several approaches. All approaches produced comparable results, and the data generated by the different approaches were significantly correlated. Variability among estimates of HC₅ of all inclusive species decreased with increasing sample size, and variability was similar among the statistical methods applied. Of the statistical methods selected, the bootstrap method represented the best-fitting model for all chemicals, while log-triangle and Weibull were the best models among the parametric methods evaluated. The bootstrap method was the primary choice to derive water quality criteria when data points are sufficient (more than 20). If the available data are few, all other methods should be constructed, and that which best describes the distribution of the data was selected.     

Toxicity of prednisolone, dexamethasone and their photochemical derivatives on aquatic organisms

Light exposure of aqueous suspensions of prednisolone and dexamethasone causes their partial phototransformation. The photoproducts, isolated by chromatographic techniques, have been identified by spectroscopic means. Prednisolone, dexamethasone and their photoproducts have been tested to evaluate their acute and chronic toxic effects on some freshwater chain organisms. The rotifer Brachionus calyciflorus and the crustaceans Thamnocephalus platyurus and Daphnia magna were chosen to perform acute toxicity tests, while the alga Pseudokircheneriella subcapitata (formerly known as Selenastrum capricornutum) and the crustacean Ceriodaphnia dubia to perform chronic tests. The photochemical derivatives are more toxic than the parent compounds. Generally low acute toxicity was found. Chronic exposure to this class of pharmaceuticals caused inhibition of growth population on the freshwater crustacean C. dubia while the alga P. subcapitata seems to be less affected by the presence of these drugs.     

Statistical evaluation of chronic toxicity data on aquatic organisms for the hazard identification: the chemicals toxicity distribution approach

Presently, in the Globally Harmonised System of Classification and Labelling of Chemicals the classification of substances for long-term effects to aquatic life is based on acute toxicity in combination with degradation and/or bioaccumulation potential. Recently an OECD Working Group was created to develop the classification scheme to accommodate chronic toxicity data related to aquatic organisms for assigning a chronic hazard category. This study focuses on a new approach for setting chronic toxicity cut-off values based on Chemicals Toxicity Distributions (CTDs). A CTD is obtained through statistical fitting of the data used by regulatory bodies for setting hazard-based classifications. The CTDs were made using the lowest aquatic NOEC value of each chemical. A review of different toxicological sources reporting acute aquatic toxicities was carried out. Initially, the data were arranged according to the specific source and distributions for key taxonomic groups (i.e. fishes, crustaceans and algae) were evaluated separately. In most cases, no significant departures from normality were observed. Thereafter, a compiled database containing >900 values was developed and the CTDs were constructed for each taxonomic group. Significant deviation from normality (P < 0.05) was observed in the fishes and crustaceans' CTDs. However, this deviation was apparently produced by the presence of only seven values with NOECs <1 x 10(-5) mg l(-1), while high correlation between the data and the normal scores (r-values>or= 0.989) indicated that the data were samples from normal distributions. From these observations, potential cut-off values would allow quantitative estimations of the percentage of chemicals falling into each specific category. This approach results in a simple classification hazard scheme where most chemicals are covered in one of the categories, allowing a clear distribution of the chemicals among three categories for chronic toxicity.     

Review of historical aquatic toxicity and bioconcentration data for the brominated flame retardant tetrabromobisphenol A (TBBPA): effects to fish,invertebrates, algae,and microbial communities

This paper summarizes the historical and recent research on the aquatic toxicology and bioconcentration potential of tetrabromobisphenol A (TBBPA), a major flame retardant in electronics. Historical studies on TBBPA are presented in detail, and are compared with more recent research. The historical studies have not been published to date, though they were pivotal in regulatory assessments by the European Union, Canada, and the USA. These assessments have enabled the use of TBBPA as a flame retardant in electronic applications, to the present. The studies were conducted under a Test Rule by the US Environmental Protection Agency in 1987, and were sponsored by member companies of the North American Flame Retardants Alliance (NAFRA) through the American Chemistry Council. The studies were conducted under Good Laboratory Practice procedures, and include 6 acute toxicity tests of TBBPA with fish, invertebrates, algae, and microbes, eight chronic tests, and three bioconcentration studies with fish and invertebrates. Methods and empirical data for each study are detailed in an electronic supplement. Results of the NAFRA studies are compared with recent findings on TBBPA toxicity. Molluscan shell growth may be uniquely sensitive to TBBPA, more sensitive than chronic fish or crustacean toxicity endpoints. Several of the NAFRA studies and several independent studies have reported toxicities exceeding the empirical water solubility limits of TBBPA (in the range of 2.0 mg/L depending on pH). The validity of these results is discussed.     

A biology-based dynamic approach for the reconciliation of acute and chronic toxicity tests: application to Daphnia magna

There is the need to integrate existing toxicity data in a coherent framework for extending their domain of applicability as well as their extrapolation potential. This integration would also reduce time and cost-consuming aspects of these tests and reduce animal usage. In this work, based on data extracted from literature, we have assessed the advantages that a dynamic biology-toxicant fate coupled model for Daphnia magna could provide when assessing toxicity data, in particular, the possibility to obtain from short-term (acute) toxicity test long-term (chronic) toxicity values taking into account the inherent variability of D. magna populations and the multiple sources of data. The results show that this approach overcomes some of the limitations of existing toxicity tests and that the prediction errors are considerably reduced when compared with the factor from 2 to 5 obtained using acute-to-chronic ratios.     

Toxicity of hexahydro-1,3,5-trinitro-1,3,5-triazine to larval zebrafish (Danio rerio)

Hexahydro-1,3,5-trinitro-1,3,5-triazine, a cyclonitramine commonly known as RDX, is used in the production of military munitions. Contamination of soil, sediment, and ground and surface waters with RDX has been reported in different places around the world. Acute and subacute toxicities of RDX have been relatively well documented in terrestrial vertebrates, but among aquatic vertebrates the information available is limited. The objective of this study was to characterize the acute toxicity of RDX to larval zebrafish. Mortality (LC50) and incidence of vertebral column deformities (EC50) were two of the end points measured in this study. The 96-h LC50 was estimated at 22.98 and 25.64 mgl(-1) in two different tests. The estimated no-observed-effective-concentration (NOEC) values of RDX on lethality were 13.27+/-0.05 and 15.32+/-0.30 mgl(-1); and the lowest-observed-effective-concentration (LOEC) values were 16.52+/-0.05 and 19.09+/-0.23 mgl(-1) in these two tests, respectively. The 96-h EC50 for vertebral deformities on survivors from one of the acute lethality tests was estimated at 20.84 mgl(-1), with NOEC and LOEC of 9.75+/-0.34 and 12.84+/-0.34 mgl(-1), respectively. Behavioral aberrations were also noted in this acute toxicity study, including the occurrence of whirling movement and lethargic behavior. The acute effects of RDX on survival, incidence of deformities, and behavior of larval zebrafish occurred at the high end of the most frequently reported concentrations of RDX in aquatic environments. The chronic effects of RDX in aquatic vertebrates need to be determined for an adequate assessment of the ecological risk of environmental RDX.     

Acute and Chronic Toxicity of Chlorine Dioxide (ClO2) and Chlorite (ClO2ˉ) to Rainbow Trout (Oncorhynchus mykiss) (4 pp)

Goal, Scope and Background Chlorite (ClO2ˉ) is a primary decomposition product when chlorine dioxide (ClO2) is added during water treatment; therefore the toxic effects of both compounds on aquatic organisms are possible. Limited data are available concerning their toxicity to fish. The aim of this study was to investigate sensitivity of rainbow trout to acute and chronic toxicity of chlorine dioxide and chlorite, and to estimate the Maximum-Acceptable-Toxicant-Concentration (MATC) of those compounds in fish. Methods The acute and chronic toxicity of chlorine dioxide and chlorite to larval and adult rainbow trout was investigated in 96-hour to 20-day laboratory exposures evaluating the wide range spectrum of biological indices under semi-static conditions. Results and Discussion Median lethal concentration (96-hour LC50) values derived from the tests were: 2.2 mg/l for larvae; 8.3 mg/l for adult fish and 20-day LC50 for larvae was 1.6 mg/l of chlorine dioxide, respectively. Chlorite was found to be from 48 to 18 times less acutely toxic to larvae and adult fish, correspondingly. Both chemical compounds induced similar toxic effects in rainbow trout larvae during chronic tests (they affected cardio-respiratory and growth parameters), but chlorine dioxide had a higher toxic potency than chlorite. A significant decrease in the heart rate and respiration frequency of larvae was established. However, within an increase in exposure duration recovery of cardio-respiratory responses was seen to have occurred in larvae exposed to chlorite. Meanwhile, in larvae exposed to chlorine dioxide, a significant decrease in cardio-respiratory responses remained during all 20-day chronic bioassays. Chlorine dioxide also more strongly affected growth parameters of rainbow trout larvae at much lower test concentrations. Decreased rate of yolk-sack resorption occurred only in the tests with chlorine dioxide. Conclusions Maximum-Acceptable-Toxicant-Concentration (MATC) of 0.21 mg/l for chlorine dioxide and of 3.3 mg/l for chlorite to fish was derived from chronic tests based on the most sensitive parameter of rainbow trout larvae (growth rate). According to substance toxicity classification accepted for Lithuanian inland waters, chlorine dioxide and chlorite can be referred to substances of \moderate\ toxicity to fish. Recommendations and Outlook Due to its very reactive nature, chlorine dioxide is rapidly (in a few hours) reduced to chlorite, which is persistent also as a biocide but 16 times less toxic to fish, according to MATC. Therefore, it is much more likely that fish will be exposed to chlorite than to chlorine dioxide in natural waters. Presently accepted, the Maximum-Permitted-Concentration of total residual chlorine (TRC) in waste-water discharging into receiving waters is 0.6 mg/l. If this requirement will not be exceeded, it is unlikely that fish would be exposed to lethal or even to sublethal concentrations of chlorine dioxide or chlorite. Furthermore, chlorine dioxide does not generate toxic nitrogenous (chloramines) or carcinogenic organic residuals (trihalomethanes). All these properties make chlorine dioxide a more promising biocide than chlorine.     

Deriving freshwater quality criteria for 2,4-dichlorophenol for protection of aquatic life in China

Freshwater quality criteria for 2,4-dichlorophenol (2,4-DCP) were developed with particular reference to the aquatic biota in China, and based on USEPA's guidelines. Acute toxicity tests were performed on nine different domestic species indigenous to China to determine 48-h LC50 and 96-h LC50 values for 2,4-DCP. In addition, 21 day survival-reproduction tests with Daphnia magna, 30-day embryo-larval tests with Carassius auratus, 60 day fry-juvenile test with Ctenopharyngodon idellus, 30 d early life stage tests with Bufo bufo gargarizans and 96 h growth inhibition tests with Scenedesms obliqaus were conducted, to estimate lower chronic limit (LCL) and upper chronic limit (UCL) values. The final acute value (FAV) was 2.49 mg/l 2,4-DCP. Acute-to-chronic ratios (ACR) ranged from 3.74 to 22.5. The final chronic value (FCV) and the final plant value (FPV) of 2.4-DCP were 0.212 mg/l and 7.07 mg/l respectively. Based on FAV, FCV, and FPV, a criteria maximum concentration (CMC) of 1.25 mg/l and a criterion continuous concentration (CCC) of 0.212 mg/l were derived. The results of this study provide useful data for deriving national or local water quality criteria for 2,4-DCP based on aquatic biota in China.     

Sensitivity of hypogean and epigean freshwater copepods to agricultural pollutants

Widespread pollution from agriculture is one of the major causes of the poor freshwater quality currently observed across Europe. Several studies have addressed the direct impact of agricultural pollutants on freshwater biota by means of laboratory bioassays; however, as far as copepod crustaceans are concerned, the ecotoxicological research is scarce for freshwater species and almost nonexistent for the hypogean ones. In this study, we conducted a comparative analysis of the available literature data on the sensitivity of freshwater copepods to agricultural pollutants. We also assessed the acute and chronic sensitivity of a hypogean and an epigean species, both belonging to the Crustacea Copepoda Cyclopoida Cyclopidae, to two N-fertilizers (urea and ammonium nitrate) and two herbicides (ARIANE^TM II from Dow AgroSciences LLC, and Imazamox), widely used for cereal agriculture in Europe. According to the literature review, freshwater copepods are sensitive to a range of pesticides and N-fertilizers. Ecotoxicological studies on hypogean species of copepods account only one study. There are no standardized protocols available for acute and chronic toxicity tests for freshwater copepods, making comparisons about sensitivity difficult. From our experiments, ionized ammonia proved to be more toxic than the herbicide Imazamox, in both short and chronic bioassays. Urea was the less toxic chemical for both species. The hypogean species was more sensitive than the epigean one to all chemicals. For both species and for all tested chemicals, acute lethality and chronic lethality were induced at concentrations higher than the law limits of good water body quality in Europe, except for ionized ammonia, which provoked the chronic lethality of the hypogean species at a lower concentration. The hazardous concentration (HC) of un-ionized ammonia for 5 % of freshwater copepods, obtained by a species sensitivity distribution, was 92 μg l^?1, significantly lower than the HC computed for traditional test species from freshwater environments.     

Multi-generation cadmium acclimation and tolerance in Daphnia magna Straus

The cladoceran Daphnia magna was acclimated for seven generations to cadmium concentrations ranging from 0 (control) to 250 microg/l Cd (corresponding to a free ion activity of 4.60 nM Cd2+). Acute and chronic cadmium tolerance as well as cadmium accumulation were monitored as a function of acclimation time. After two to three generations of acclimation to concentrations ranging from 0.23 to 1.11 nM Cd2+ increases in acute tolerance were maximal (factor 7.2) and significant. Acclimation for seven generations to the same acclimation concentrations did result in an increased chronic cadmium tolerance (21 days EC50 values increased). Organisms acclimated to 1.93 nM Cd2+ were equally or more sensitive than non-acclimated daphnids in acute and chronic toxicity tests. Cadmium contents in D. magna increased significantly as a function of the acclimation concentration. Maximum body burdens of 236+/-30 microg Cd/g dry weight were measured in organisms exposed to 4.60 nM Cd2+, but detoxification mechanisms were only successful up to 82+/-20 microg Cd/g dry weight as this concentration did not cause major decreases in survival and reproduction in chronic toxicity tests. As the potential positive effect of acclimation on cadmium tolerance disappeared with successive acclimation generations and increasing acclimation concentrations, it is concluded that multi-generation acclimation studies are important for the evaluation of the long-term effects of environmental toxicants.     

Comparing aquatic risk assessment methods for the photosynthesis-inhibiting herbicides metribuzin and metamitron

Three different risk assessment procedures are described that aim to protect freshwater habitats from risks of the photosynthesis-inhibiting herbicides metribuzin and metamitron. These procedures are (1) the first-tier approach, based on standard toxicity tests and the application of an assessment factor, (2) the Species Sensitivity Distribution (SSD) approach, based on laboratory tests with a wider array of species and the application of a statistical model to calculate the HCx (the Hazardous Concentration for x% of the species), and (3) the model ecosystem approach, based on the evaluation of treatment-related effects in field enclosures. A comparison of the risk assessment procedures reveals that the first-tier approach is the most conservative for metamitron and metribuzin, and that HC5 values (and even HC10 values) based on acute EC50 values of algae and aquatic vascular plants may be used to derive maximum permissible concentrations for single applications. For both compounds these HC5 values were very similar to the ecological threshold concentrations in the enclosure studies. In contrast to model ecosystem experiments, however, HCx values based on lab toxicity tests do not provide information on the recovery potential of sensitive endpoints and on indirect effects, which may be important for regulatory decision-making. In the enclosure study, indirect effects of metribuzin on invertebrate populations were observed at an exposure concentration that was approximately 20 times lower than the corresponding HC5 value based on lab toxicity data for aquatic invertebrates.     

Small-scale variability of metals in soil and composite sampling

Soil pollution data is also strongly scattering at small scale. Sampling of composite samples, therefore, is recommended for pollution assessment. Different statistical methods are available to provide information about the accuracy of the sampling process. Autocorrelation and variogram analysis can be applied to investigate spatial relationships. Analysis of variance is a useful method for homogeneity testing. The main source of the total measurement uncertainty is the uncertainty arising from sampling. The sample mass required for analysis can also be estimated using an analysis of variance. The number of increments to be taken for a composite sample can be estimated by means of simple statistical formulae. Analytical results of composite samples obtained from different fusion procedures of increments can be compared by means of multiple mean comparison. The applicability of statistical methods and their advantages are demonstrated for a case study investigating metals in soil at a very small spatial scale. The paper describes important statistical tools for the quantitative assessment of the sampling process. Detailed results clearly depend on the purpose of sampling, the spatial scale of the object under investigation and the specific case study, and have to be determined for each particular case.     

Evaluation of air quality zone classification methods based on ambient air concentration exposure

Air quality zones are used by regulatory authorities to implement ambient air standards in order to protect human health. Air quality measurements at discrete air monitoring stations are critical tools to determine whether an air quality zone complies with local air quality standards or is noncompliant. This study presents a novel approach for evaluation of air quality zone classification methods by breaking the concentration distribution of a pollutant measured at an air monitoring station into compliance and exceedance probability density functions (PDFs) and then using Monte Carlo analysis with the Central Limit Theorem to estimate long-term exposure. The purpose of this paper is to compare the risk associated with selecting one ambient air classification approach over another by testing the possible exposure an individual living within a zone may face. The chronic daily intake (CDI) is utilized to compare different pollutant exposures over the classification duration of 3 years between two classification methods. Historical data collected from air monitoring stations in Kuwait are used to build representative models of 1-hr NO₂ and 8-hr O₃ within a zone that meets the compliance requirements of each method. The first method, the “3 Strike” method, is a conservative approach based on a winner-take-all approach common with most compliance classification methods, while the second, the 99% Rule method, allows for more robust analyses and incorporates long-term trends. A Monte Carlo analysis is used to model the CDI for each pollutant and each method with the zone at a single station and with multiple stations. The model assumes that the zone is already in compliance with air quality standards over the 3 years under the different classification methodologies. The model shows that while the CDI of the two methods differs by 2.7% over the exposure period for the single station case, the large number of samples taken over the duration period impacts the sensitivity of the statistical tests, causing the null hypothesis to fail. Local air quality managers can use either methodology to classify the compliance of an air zone, but must accept that the 99% Rule method may cause exposures that are statistically more significant than the 3 Strike method.

Implications: A novel method using the Central Limit Theorem and Monte Carlo analysis is used to directly compare different air standard compliance classification methods by estimating the chronic daily intake of pollutants. This method allows air quality managers to rapidly see how individual classification methods may impact individual population groups, as well as to evaluate different pollutants based on dosage and exposure when complete health impacts are not known.     

Toxic effects of orimulsion on rainbow trout <Emphasis Type="Italic">oncorhynchus mykiss</Emphasis>

GOAL, SCOPE AND BACKGROUND: Orimulsion (stable emulsion of natural bitumen and water) is a new imported industrial fuel in Lithuania. No data on its toxicity to fish is freely available. The aim of this study was to investigate sensitivity of rainbow trout (Oncorhynchus mykiss) to acute and chronic toxicity of orimulsion and to estimate the Maximum Acceptable Toxicant Concentration (MATC) of orimulsion to fish. METHODS: Laboratory tests were conducted on rainbow trout in all stages of development (embryos, larvae, adults). Acute toxicity (96-hour duration) and long-term (28 or 60-day duration) tests evaluating the wide range spectrum of biological indices were performed under semi-static conditions. RESULTS AND DISCUSSION: Median lethal concentration (96-hour LC50) values and their 95% confidence intervals derived from the tests were: 0.1 (0.09-0.12) to embryos, 0.06 (0.05-0.07) to larvae and 2.22 (2.02-2.43) to adult fish, and 28-day LC50 to adult fish was found to be 0.26 (0.21-0.32) g/l of total orimulsion respectively. The acute toxicity of orimulsion to rainbow trout can be characterised by a narrow zone of toxic effect and a sharp boundary between lethal and sublethal concentrations. The lowest 'safe' or 'no-effect' concentration values of total orimulsion obtained in long-term tests were equal to 0.09 g/l to adult fish, 0.019 g/l to embryos, and 0.0017 g/l to larvae. Proposed value of 'application factor' for orimulsion was found to be equal to 0.03. Since orimulsion has the property to disperse in all water volume, its toxic effect on fish can be characterised by the combined effects of dispersion and water-soluble-fraction. CONCLUSIONS: Maximum Acceptable Toxicant Concentration (MATC) of 0.0017 g/l of total orimulsion to fish was derived from long-term tests based on the most sensitive parameter of rainbow trout larvae (relative mass increase at the end of the test). According to substance toxicity classification accepted for Lithuanian inland waters, orimulsion can be referred to substances of 'moderate' toxicity to fish. RECOMMENDATIONS AND OUTLOOK: For prediction and evaluation of toxic impact of orimulsion accident spills on fish, some recommendations should be given. Since orimulsion has the property to disperse in all water volume during short time periods, the amounts of both spilled orimulsion and polluted water should be ascertained. Once both parameters are known, the real concentration of orimulsion in the water body must be determined. Then this concentration must be compared with 'safe' concentration to fish. By use of 'application factor' 0.03, approximate MATC for other fish species can be estimated when only acute toxicity data (96-hour LC50 value) is available.     

Statistical Methods and Pitfalls in Environmental Data Analysis

This paper reviews four commonly used statistical methods for environmental data analysis and discusses potential pitfalls associated with application of these methods through real case study data. The four statistical methods are percentile and confidence interval, correlation coefficient, regression analysis, and analysis of variance (ANOVA). The potential pitfall for estimation of percentile and confidence interval includes the automatic assumption of a normal distribution to environmental data, which so often show a log-normal distribution. The potential pitfall for correlation coefficient includes the use of a wide range of data points in which the maximum in value may trivialize other smaller data points and consequently skew the correlation coefficient. The potential pitfall for regression analysis includes the propagation of uncertainties of input variables to the regression model prediction, which may be even more uncertain. The potential pitfall for ANOVA includes the acceptance of a hypothesis as a weak argument to imply a strong conclusion. As demonstrated in this paper, we may draw very different conclusions based on statistical analysis if the pitfalls are not identified. Reminder and enlightenment obtained from the pitfalls are given at the end of this article.     

Statistical Methods and Pitfalls in Environmental Data Analysis

This paper reviews four commonly used statistical methods for environmental data analysis and discusses potential pitfalls associated with application of these methods through real case study data. The four statistical methods are percentile and confidence interval, correlation coefficient, regression analysis, and analysis of variance (ANOVA). The potential pitfall for estimation of percentile and confidence interval includes the automatic assumption of a normal distribution to environmental data, which so often show a log-normal distribution. The potential pitfall for correlation coefficient includes the use of a wide range of data points in which the maximum in value may trivialize other smaller data points and consequently skew the correlation coefficient. The potential pitfall for regression analysis includes the propagation of uncertainties of input variables to the regression model prediction, which may be even more uncertain. The potential pitfall for ANOVA includes the acceptance of a hypothesis as a weak argument to imply a strong conclusion. As demonstrated in this paper, we may draw very different conclusions based on statistical analysis if the pitfalls are not identified. Reminder and enlightenment obtained from the pitfalls are given at the end of this article.     

Aquatic predicted no-effect concentration for three polycyclic aromatic hydrocarbons and probabilistic ecological risk assessment in Liaodong Bay of the Bohai Sea, China

Predicted no-effect concentration (PNEC) is often used in ecological risk assessment to determine low-risk concentrations for chemicals. In the present study, native marine species were selected for toxicity testing. The PNECs for three polycyclic aromatic hydrocarbons (PAHs), specifically phenanthrene (Phe), pyrene (Pyr), and benzo[a]pyrene (BaP), were derived from chronic and acute toxicity data with log-normal statistical methods. The achieved PNECs for Phe, Pyr, and BaP were 2.33, 1.09, and 0.011 μg/L, respectively. In Jinzhou Bay and the Shuangtaizi River Estuary of Liaodong Bay in the Bohai Sea, China, the surface water concentrations of the three PAHs were analyzed by gas chromatography–mass spectrometry. Based on two probabilistic ecological risk assessment (PERA) methods, namely probabilistic risk quotient and joint probability curve, the potential risk of Phe, Pyr, and BaP in Jinzhou Bay and Shuangtaizi River Estuary was assessed. The same order of ecological risk (BaP > Phe > Pyr) was found by both models. Our study considered regional characteristics of marine biota during the calculation of PNECs, and the PERA methods provided probabilities of potential ecological risks of chemicals. Within the study area, further research on BaP is required due to its high potential ecological risk.     

Avoidance behaviour of Enchytraeus albidus: effects of benomyl, carbendazim, phenmedipham and different soil types

Enchytraeids are typical inhabitants of many soils, contributing to vital processes of this environmental compartment. Indirectly they are involved in regulating the degradation of organic matter, as well as improving the pore structure of the soil. Due to their behaviour, they are able to avoid unfavourable environmental conditions. Avoidance tests with enchytraeids, initially developed with earthworms by several authors, are quick and easy to perform. With these tests a first assessment of the toxicity of a (contaminated or spiked) soil is possible in just 48 h by using the reaction of the enchytraeids as measurement endpoint. In this period of time the organisms can choose between the control soil and the other soil (a contaminated or spiked or another soil with different physico-chemical properties). In the tests reported here, the enchytraeids were exposed to control soils spiked with the fungicides Benomyl and Carbendazim and the herbicide Phenmedipham. Several chemical concentrations were tested in order to evaluate the avoidance behaviour to toxic substances. In fact, often these short-term screening tests gave results showing avoidance at concentrations in a range similar to the acute test results but, higher than in chronic tests. Further tests are needed to decide whether the results gained in this study can be extrapolated to other chemicals. It is proposed to standardize the Enchytraeid Avoidance Test as it is currently done for the Earthworm Avoidance Test by the International Standard Organization (ISO).     

Investigation of acute toxicity of beta-cypermethrin on guppies Poecilia reticulata

Beta-cypermethrin, a synthetic pyrethroid pesticide and potential toxic pollutant, contaminating aquatic ecosystems was investigated in the present study for acute toxicity. Guppy fish (Poecilia reticulata) was selected for the bioassay experiments. The experiments were repeated three times and the 48-h LC50 was determined for the guppies. The static test method of acute toxicity test was used. Water temperature was regulated at 22 +/- 1 degrees C. In addition, behavioral changes at each beta-cypermethrin concentration were observed for the individual fish. Data obtained from the beta-cypermethrin acute toxicity tests were evaluated using the probit analysis statistical method. The 48-h LC50 value for guppy was estimated as 21.4 microg/l.