Histological alternation and vitellogenin induction in adult rare minnow (Gobiocypris rarus) after exposure to ethynylestradiol and nonylphenol

Adult rare minnow (Gobiocypris rarus) were exposed to 0, 1, 5, and 25 ng/l (nominal concentrations) of 17alpha-ethynylestradiol (EE2) and 3, 10, and 30 microg/l (nominal concentrations) of 4-nonylphenol (NP) under flow-through conditions for a period of 28 d. Low mortality was observed at 5 and 25 ng/l EE2 and the growth of fish reduced significantly at 25 ng/l EE2 compared to controls. However, the gonadosomatic indices (GSI) of male fish were significantly higher in 1 ng/l EE(2) treatments and in 10 and 30 microg/l NP treatments (p<0.05). Renal somatic indices (RSI) of male fish in EE2 treatments were significantly higher than those in controls (p<0.05). In contrast, significantly decreased GSI and RSI of female fish could only be observed in 5 and 25 ng/l EE(2) treatments (p<0.05). Hepatosomatic indices (HSI) of male fish were significantly higher in 25 ng/l EE(2) treatments. However, significantly increased of HSI of female fish could only be observed in 1 ng/l EE(2) treatments. Plasma vitellogenin (VTG) induction could be observed in males after exposed to different concentrations of EE2 and NP, and plasma VTG concentrations in females exposed to 5 and 25 ng/l EE(2) were also significantly higher than in controls (p<0.05). At level higher than 5 ng/l EE2 or 30 microg/l NP, hepatic tissue and renal tissue impairment of males could be observed. The pathological male liver was associated with a hypertrophy of hepatocytes and damages to cellar structure and accumulated eosinophilic material. Renal tissue showed different pathological effects which was reflected by accumulated eosinophilic material, hemorrhages within the kidney tubules and hypertrophy of the tubular epithelia. Also at these levels of exposure, feminization of male fish could be noticed and parts of males manifested the testis-ova phenomenon. Ovaries of female rare minnow in 25 ng/l EE2 treatment group were degenerated. Therefore when exposed to EE2 and NP even at environmental observed concentrations, adverse effects could occur in the reproductive system of adult fishes. The observed hepatic tissue and renal tissue impairment should be due to the induction and accumulation of VTG in organs, especially in males.     

Effects of waterborne exposure to 4-nonylphenol on plasma sex steroid and vitellogenin concentrations in sexually mature male carp (Cyprinus carpio)

4-Nonylphenol (NP) has been shown to elicit estrogenic responses both in vivo and in vitro. The mechanism by which NP exerts estrogenic and other endocrine-modulating effects in vivo remains unclear, however. The goal of this study was to evaluate the ability of NP to elicit estrogenic responses through indirect mechanisms of action involving the modulation of endogenous steroid hormone concentrations. Sexually mature male common carp (Cyprinus carpio) were exposed to aqueous NP concentrations ranging from <0.05 to 5.4 microg NP/l for 28-31 d. Approximately 0.5-3.5 ppm of NP was detected in pooled plasma samples or tissue samples from the carp studied. NP exposure did not significantly increase plasma concentrations of 17beta-estradiol (E2), testosterone (T) or vitellogenin (VTG). Excluding outliers, plasma E2 concentrations ranged from <175 to 700 pg E2/ml. T concentrations ranged from 940 to 24,700 pg T/ml plasma. The greatest VTG concentration detected was 52 microg/ml. One-third of the plasma samples tested contained <1 microg VTG/ml. Overall, the results of this study did not support the hypothesis that exposure to waterborne NP can modulate concentrations of steroid hormones in the plasma of sexually mature male carp. The results did, however, raise a number of questions regarding the utility of estradiol equivalent (EEQ) estimates as a means of predicting in vivo effects of estrogenic substances. Furthermore, they provide information regarding the concentrations and variability of E2, T, and VTG in the plasma of sexually mature male carp, which may aid in design and interpretation of future studies.     

Acute and chronic toxicities of tributyltin to various life stages of the marine polychaete Hydroides elegans

Experiments were performed to determine the acute and chronic toxicities of tributyltin (TBT) (nominal concentrations) to polychaete Hydroides elegans. Firstly, static tests were performed to examine the lethal effect of TBT on five life stages of H. elegans. The 48-h LC(50) (95% CI) for the eggs, 2-cells, trochophores, juveniles and adults was 0.18 (0.15-0.20), 0.97 (0.77-1.23), 2.36 (2.11-2.65), 2.86 (2.63-3.12) and 4.36 (4.04-4.71) microg TBT l(-1), respectively. Secondly, a 17-d static renewal chronic test was conducted to study sensitivity of the early development (egg to juvenile) to TBT. The survivorship, percent settlement and time to reach settlement were significantly reduced at 0.01 microg TBT l(-1). Effects of TBT on the juvenile growth and maturation were also studied through a 44-d chronic test. Lower survivorship and reduced tube growth were observed only at >or= 1 microg TBT l(-1), however, only H. elegans exposed to or= 0.1 microg TBT l(-1). Our results indicate that the early development of H. elegans is highly sensitive to TBT and this polychaete can be routinely employed as a test organism for both acute and chronic ecotoxicity bioassays in tropical and subtropical regions such as southern China.     

Developmental effects in Japanese medaka (Oryzias latipes) exposed to nonylphenol ethoxylates and their degradation products

The endocrine modulating potency of five alkylphenol compounds to fish, including nonylphenol (NP), three nonylphenol ethoxylate mixtures (NP1EO, NP4EO, NP9EO) and one nonylphenol ethoxycarboxylate (NP1EC) was assessed using in vivo tests conducted with Japanese medaka (Oryzias latipes). Medaka exposed to test materials from 1 day to 100 days post-hatch were monitored for alterations to sex ratios and secondary sex characteristics and development of gonadal intersex (i.e., testis-ova). The treatment with 100 microg l-1 NP (measured 29 microg l-1) induced gonadal intersex in over 80% of exposed males, mixed secondary sex characteristics in over 40% of exposed fish and suppression of the development of papillae on the anal fin of 100% of males. The 30 microg l-1 NP (measured 8.7 microg l-1) treatment induced gonadal intersex in only one of the 22 exposed males and mixed secondary sex characteristics in approximately 20% of the exposed fish. An elevated incidence of fish with mixed secondary sex characteristics and suppression of papillae development was also observed in the treatment with NP1EO at the highest test concentration of 300 microg l-1 (measured 105 microg l-1). There was no evidence of mixed secondary sex characteristics or gonadal intersex in treatments with the remaining test mixtures. This study confirms that NP is an estrogenic compound that could affect gonadal development in fish chronically exposed to concentrations in the range of 10 microg l-1. NP1EO is very weakly estrogenic at concentrations that are an order of magnitude higher than the lowest observed effect concentration for nonylphenol.     

Morphological effects of Bisphenol-A on the early life stages of medaka (Oryzias latipes).

Bisphenol-A (BPA), a widely used polycarbonate plasticizer, has been of concern because it has been shown to leach out of plastics and other epoxy products. Primary sources of environmental releases are expected to be from BPA and epoxy manufacturing facilities. Although environmental concentrations may be limited, little is known about the effects of this compound on fish, particularly during their most sensitive early life stages. A pilot study exposing newly hatched medaka to 200 microg BPA/l for 96 h showed to be non-lethal. In addition, medaka embryos were exposed beginning at 5 h post-fertilization, for 9 days at 25 degrees C, to concentrations of 20 or 200 microg BPA/l (24 h static renewal). Embryos were monitored daily for development and gross abnormalities. Embryos exposed to 200 microg BPA/l did not exhibit abnormalities until after day 4. However, between days 4 and 8 the severity index (SI) score of embryos was significantly greater than those exposed to lesser concentrations. By day 9, SI scores were not statistically different among treatments. BPA caused transient embryonic deformities in medaka at environmentally relevant concentrations.     

Nonylphenolic compounds in drinking and surface waters downstream of treated textile and pulp and paper effluents: a survey and preliminary assessment of their potential effects on public health and aquatic life

Eleven drinking water treatment plants, located downstream of textile plants or pulp and paper mills, have been sampled monthly during a year for the analysis of 17 nonylphenol ethoxylates (NP1-17EO) and two nonylphenoxycarboxylic acids (NP1-2EC). At all but one plant, results in the drinking water, for the sum of these 19 substances, range between below detection levels and 6.7 microg/l. Annual means are between 0.02 and 2.8 microg/l. At the other plant, the yearly average concentration is 10.4 microg/l and the monthly maximum is 43.3 microg/l. In the surface (pre-treatment) water, the annual mean concentrations of the 11 plants range between 0.14 and 17.8 microg/l and the recorded instantaneous maximum is 55.3 microg/l. According to Canadian health authorities, drinking water is a negligible route of human exposure to nonylphenolic compounds, even at the highest concentrations found in this study. After transformation of the data into nonylphenol equivalents, about 20% of the surface water samples exceed the Canadian 1 microg/l nonylphenol water quality guideline for the protection of aquatic life. Some results also exceed Québec's 6 microg/l nonylphenol guideline. The efficiency of the plants in removing nonylphenolic compounds from drinking water is highly variable, ranging from 11% to 99%.     

Nitrophenols in precipitation

From 1995 to 1998 the concentrations of 4-nitrophenol, 2-methyl-4-nitrophenol, 3-methyl-4-nitrophenol, dinitro-ortho-cresol (DNOC) and 2,4-di-nitrophenol were measured monthly by HPLC in precipitation at eight different locations in Bavaria (Germany). Samples were collected by purpose-constructed computerised rainwater samplers which record electronically various sensor data each hour and adjust the sample temperature to 4 degrees C. The highest nitrophenol (NP) concentrations were measured for 4-NP. The median at all locations is higher than 1 microg/l. The median of the other NPs ranges between 0.2 and 0.8 microg/l. Considering the rain amounts the highest depositions were calculated for the regions Spessart, Bayerischer Wald and Chiemgauer Alpen. The median of 4-NP depositions extents to 200 microg/(month m2). The highest medians of the other NP depositions reach approximately 50 microg/(month m2).     

Effects of chlorpyrifos on reproductive performances of guppy (Poecilia reticulata)

Guppy (Poecilia reticulata) was selected to investigate the effects of chlorpyrifos on reproductive performances. Male and female guppy with proven fertility were selected from our own colony and the groups of fish (n=72/group) were exposed to pre-determined chlorpyrifos concentrations (0.002 microg/l, 2 microg/l) based on the 96-h LC50 for guppy. Mating behavior of males was recorded on the 2nd day of exposure. Offspring were counted and survival recorded on the 14th day. Gonopodial thrusts (8/15 min) in 0.002 microg/l and (4/15 min) in 2 microg/l were significantly different from the control group (11/15 min). Similarly, live birth reduced significantly to 8/female in 2 microg/l compared to 27/female in the control group. Survival of offspring after 14 days was reduced to 47% in the 2 microg/l group compared to 94% of survival in the control. Our findings demonstrate that low soluble concentrations of chlorpyrifos affect mating behavior, number of offspring and offspring survival of guppy.     

Bisphenol A induces yolk-sac oedema and other adverse effects in landlocked salmon (Salmo salar m. sebago) yolk-sac fry

Accumulation and toxicity of waterborne bisphenol A were studied in landlocked salmon (Salmo salar m. sebago) yolk-sac fry. In a short-term (96 h) exposure to five bisphenol A concentrations yolk-sac fry had higher accumulation rates and bioconcentration factors (BCF96) than earlier studies have shown for salmon eggs. Furthermore, the conditional uptake rate constant tended to decrease as exposure concentration increased. Fry were also exposed to bisphenol A for 42 days at three concentrations (10, 100 and 1000 microg/l), and changes in behaviour, morphology and histological structure were observed. After 6 days of exposure, the highest concentration (1000 microg/l) of bisphenol A caused fluid accumulation (oedema) in the yolk sac and haemorrhages in the front part of the yolk sac and in the head around the gill arches. Later on, the fry at 1000 microg/l showed phlegmatic behaviour and had darker skin coloration than the fry in the other treatments. At the two highest concentrations (100 and 1000 microg/l) histological changes were seen in liver cell nuclei, where strongly stained fragments were observed. In the control fry and the fry exposed to 10 microg/l the nucleolus was clearly visible and spherical in shape and no strongly stained fragments were present. This study shows that high concentrations of bisphenol A may have both morphological and histological effects on salmon yolk-sac fry.     

Determination of nonylphenol ethoxylates and octylphenol ethoxylates in environmental samples using 13C-labeled surrogate compounds

Alkylphenol polyethoxylates (APEOs) have been widely used as nonionic surfactants in a variety of industrial and commercial products. Typical compounds are nonylphenol polyethoxylates (NPEOs) and octylphenol polyethoxylates (OPEOs), which serve as precursors to nonylphenol (NP) and octylphenol (OP), respectively. NP and 4-t-OP are known to have endocrine disrupting effects on fish (medaka, Oryzias latipes), so it is important to know the concentrations of APEOs in the environment. Because the analytical characteristics of these compounds depend on the length of the ethoxy chain, it is necessary to use appropriate compounds as internal standards or surrogates. We synthesized two 13C-labeled surrogate compounds and used these compounds as internal standards to determine NPEOs and OPEOs by high-performance liquid chromatography (LC)-mass spectrometry. Method detection limits were 0.015 microg/L for NP (2)EO to 0.037 microg/L for NP(12)EO, and 0.011 microg/L for OP(3,6)EO to 0.024 microg/L for OP (4)EO. NPEO concentrations in water from a sewage treatment plant were less than 0.05-0.52 microg/L for final effluent and 1.2-15 microg/L for influent. OPEO concentrations were less than 0.05-0.15 microg/L for the final effluent and less than 0.05-1.1 microg/L for influent.     

Bioconcentration factor of relatively low concentrations of chlorophenols in Japanese medaka

Bioconcentration factors (BCF) for pentachlorophenol (PCP) and 2,4-dichlorophenol (2,4-DCP) in Japanese medaka (Oryzias latipes) were determined at five different concentrations of the chemicals, between 0.1 and 10 microg/l (PCP), 0.3 and 30 microg/l (2,4-DCP), in the ambient water. Medaka were exposed to each chemicals in a continuous-flow system during the embryonic development period and 60 days after hatching from eggs collected in the laboratory. Both the exposure time and the aqueous concentrations are much more realistic and closer to natural aquatic environments than those used in conventional BCF studies. The BCF values of PCP were from (4.9+/-2.8)x10(3) at the aqueous concentration of 0.074+/-0.028 microg/l to (2.1+/-1.4)x10(3) at 9.70+/-0.56 microg/l. The BCF value of 2,4-DCP were from (3.4+/-3.0)x10(2) at 0.235+/-0.060 microg/l to 92+/-27 at 27.3+/-1.6 microg/l. Generally, BCF values increased as the aqueous concentrations of PCP or 2,4-DCP decreased. This finding suggests that a relatively low and realistic aqueous concentration of these compounds is necessary to more accurately determine their BCF values in natural aquatic environments. Conventional BCF experiments at higher aqueous concentrations may underestimate the BCF values.     

Distribution of alkylphenols in the Pearl River Delta and adjacent northern South China Sea, China

The occurrence of alkylphenols (APs) was investigated in surface water and sediments from the Pearl River Delta and adjacent northern South China Sea. Most of the water samples contained detectable amounts of APs, ranging up to 0.628 microg l(-1) for nonylphenol (NP) and 0.068 microg l(-1) for octylphenol (OP). APs were found in all of the sediment samples with concentrations ranging from 59 to 7808 microg kg(-1) for NP and from 1 to 93 microg kg(-1) for OP. The Zhujiang River showed the highest concentrations of APs in both water and sediments. Significant decrease of APs concentrations going from the Zhujiang River to the Shiziyang River was observed. The Xijiang River contained concentrations of APs slightly higher in water but relatively lower in sediments than the Lingding Bay, which might be attributed to their different hydrodynamic and sedimentary characteristics. There was a decreasing trend of APs in water from the rivers to the estuary and further to the sea on the whole. In the Lingding Bay and its outer waters, concentrations of APs in sediments increased to a maximum and then decrease seaward, which was consistent with the distribution trend of the sediment organic carbon contents. Linear regression analyses showed the concentrations of APs were markedly correlated with the sediment organic carbon contents, indicating that the sediment organic carbon is an important factor controlling the levels of APs in sediments.     

LC-MS-MS analysis and occurrence of octyl- and nonylphenol, their ethoxylates and their carboxylates in Belgian and Italian textile industry, waste water treatment plant effluents and surface waters

Alkylphenols (APs), alkylphenol ethoxylates (APEOs), ethoxycarboxylate metabolites (APECs) and bisphenol A were determined in surface water using solid-phase extraction (SPE) followed by triple-quadrupole LC-MS-MS. APs were separated by LC from APECs using an acetonitrile-water-gradient without the addition of any buffer. Nonylphenol ethoxycarboxylates (NPECs) interfere in the detection of nonylphenols (NPs) when using an acidic mobile phase, because they produce the same MS-MS fragment ions (219>133 and 147). 4n-NP shows the characteristic transition 219>106; it is well suited as internal standard. Nonylphenol ethoxylates NPE(n)Os (n=1-17) were analysed separately in a second run by positive ionization using an ammonium acetate mobile phase. Textile industry discharges, the corresponding wastewater treatment plant (WWTP) effluents and the receiving rivers in Belgium and Italy were analysed. Among the substances investigated, NPE1C and NPE2O exhibited the highest concentrations in the water samples, up to 4.5 microg l(-1) NPE1C in a WWTP effluent and 3.6 microg l(-1) NPE2O in a river. The highest NP levels were found in the receiving rivers (max. 2.5 microg l(-1)). The predicted no-effect concentration (PNEC) for NP of 0.33 microg l(-1) for water species was frequently exceeded in the surface waters investigated, suggesting potential adverse effects to the aquatic environment.     

Effect of tributyltin on veliger larvae of the Manila clam, Ruditapes philippinarum

We investigated the effects of waterborne and maternal exposure to tributyltin (TBT) on veliger larvae of the Manila clam, Ruditapes philippinarum. In a waterborne exposure test, veliger larvae (D-larvae stage: 24h after fertilization) were exposed to TBT at measured concentrations of <0.01 (control), 0.055, 0.130, 0.340, and 0.600microg/l for 13d. The percentage of normal veliger larvae (the ratio of normal veliger larvae to all larvae) decreased significantly in all TBT treatment groups compared with that in the control group. In a maternal exposure test, 100 clams were exposed to TBT at measured concentrations of <0.01 (control), 0.061, and 0.310microg/l at 20-22 degrees C for 3 weeks, and the percentage of normal veliger larvae assessed for 13d. No maternal effects on veliger larvae from TBT were observed in TBT treatment groups as compared with the control group. These results demonstrate that waterborne TBT affects Manila clam veliger larvae, and indicates that TBT may have reduced Manila clam populations by preventing the development and survival of veliger larvae.     

The interference of 2-chloro-5-oxo-3-hexene diacyl chloride (COHC) in the detection of strong mutagen MX

Laboratory experiments were conducted to examine the impact of cadmium (Cd) on survival of Diplostomum sp. cercariae. Freshly emerged cercariae were exposed to Cd solutions at concentrations ranging between 0.2 and 200 microg/l. The mortality patterns and mean survival times (MSTs) of the exposed parasites were determined by the Kaplan-Meier estimator and were compared (Log-Rank test) to corresponding patterns and estimators of the unexposed controls. Cd concentrations >20 microg/l caused statistically significant changes in the mortality pattern of Diplostomum sp. cercariae and reduced MST by at least five hours compared to the control group, where MST was 52 h. The results show that longevity of the free-living Diplostomum sp. cercariae can be reduced by direct exposure to heavy metals. Cd contamination of aquatic habitats thus potentially can have an impact on parasite populations and communities.     

PCB contaminated dust on indoor surfaces--health risks and acceptable surface concentrations in residential and occupational settings

Polychlorinated biphenyls (PCBs) have been used in diverse purposes such as indoor paints. Removal of these paints with dust creating techniques, like sandblasting, will result in contamination of building surfaces with PCB-containing dust. Objectives of this study was to analyze the PCB concentrations on surfaces after sandblasting with silica using wipe samples and estimate the resulting health risks and further calculate the risk based acceptable PCB surface concentrations that do not cause incremental lifetime cancer risk higher that 10(-5) or does not cause immunosupression effects in residential use or in occupational settings. Both deterministic and probabilistic approaches were used. The total PCB concentrations on surfaces ranged from 10 to 1100 microg/m(2). Estimated cancer risk was 1.2 x 10(-4) for childhood exposure, 1.3 x 10(-5) for adult residents and 1.5 x 10(-5) for occupational exposure. Probabilistic risk assessment revealed that point estimates were quite reasonable and located between 45th and 79th percentiles on probabilistic distribution of risk. The noncancer risks were calculated as hazard quotients (HQ) which ranged from 3.3 to 35 depending on the exposure scenario. Acceptable surface concentrations based on noncancer effects that are protective for 95% of exposed population were 7 microg/m(2) for residential use, 65 microg/m(2) for residential use if only adults will be exposed and 140 microg/m(2) for occupational use. Preliminary cleanup experiment revealed that when contaminated dust was carefully removed with industrial vacuum cleaner and further washed with terpene containing liquid the surface concentration dropped below the acceptable levels calculated in this study.     

4-n-Nonylphenol and 17-β estradiol may induce common DNA effects in developing barnacle larvae

There is a growing concern over the potential effects of environmental endocrine disrupters on both human and wildlife populations. However, to date, minimal research has been conducted to determine the effect of estrogens and xenoestrogens at the DNA level. In this study, we used the random amplified polymorphic DNA (RAPD) assay to evaluate the effects on the genomic DNA of barnacle larvae that had been exposed to 17β-estradiol (E₂) and low concentrations of 4-n-nonylphenol (NP). DNA effects include DNA damage as well as mutations and possibly other effects at the DNA level that can be induced by chemical or physical agents that directly and/or indirectly interact with genomic DNA. Not only did exposure to NP and E₂ induce changes in RAPD profiles in the exposed barnacle larvae when compared to control patterns, but also, and more importantly, there were similarities in the RAPD modifications in the exposed populations that had been treated to either chemical. We propose that NP and E₂ induced some common DNA effects in barnacle larvae and that these specific modifications in RAPD patterns may arise as a consequence of hot spot DNA damage (e.g. DNA adducts) and/or mutations (point mutations or genomic rearrangements). This could help to explain how xenoestrogens mimic the effects produced by natural estrogens. In conclusion, in the field of endocrine disruption, the study of DNA effects induced by estrogens and/or xenoestrogens warrants further investigation. Indeed, changes at the DNA level may be the precursors of some of the numerous effects reported at higher levels of biological organisation such as the feminization of males, developmental abnormalities, and infertility.     

Environmental chemicals with known endocrine potential affect yolk protein content in the aquatic insect Chironomus riparius

The use of vitellogenesis as a marker for possible effects of endocrine disrupting agents on insects was tested in the aquatic midge Chironomus riparius. As test substances the synthetic ecdysoid tebufenozide, and the endocrine disruptors bisphenol a and 4-n-nonylphenol were applied in a semi-static manner. The yolk protein contents of freshly emerged (24 h) male and female midges were determined by an ELISA procedure. In males, where always low amounts of immunoreactivity were apparent, yolk concentrations were lowered by 10% after a 80 microg/l tebufenozide treatment, and by 20-25% after exposition to bisphenol a at concentrations of 1, 100, and 3,000 microg/l. 4-n-nonylphenol contamination caused an inverted dose-response curve. At low test concentrations (1.9-30 microg/l) reduced yolk immunoreactivity occurred, while at medium concentrations (120 and 500 microg/l) no significant effects were observable. In the most highly contaminated group (2,000 microg/l) yolk protein immunoreactivity was elevated to 107% of the control. Female yolk protein contents were affected only in the 3,000 microg bisphenol a/l contaminated group, where yolk immunoreactivity was reduced by ca. 10% compared to the control.     

A new protocol to measure the effects of toxicants on daphnid-algae interactions

Inhibition of zooplankton grazing by toxicants present at sublethal concentrations in freshwater ecosystems can lead to uncontrolled algal growth and consequently exacerbate eutrophication problems. Short-term measurements of cladoceran grazing inhibition have been reported for some toxicants, but these studies do not mimic the actual interactions between microalgae, daphnids and toxicants, since algal growth is not allowed. On the opposite, algal blooms in complex microcosm or mesocosm assays have been interpreted as consequences on zooplankton, but effects on grazing, survival, growth or reproduction could not be easily discriminated. In this study, a simple assay with daphnids and microalgae is proposed to measure effects on grazing in dynamic conditions (algal growth over 6 days), and applied to copper and lindane. In the same time, direct effects on algal growth can be shown and taken into account. Results are compared with daphnid response measured with different endpoints (immobilization test and static grazing assay). For both toxicants, effects at sublethal concentrations were demonstrated. Copper impaired daphnid grazing at 10 microg/l (60% inhibition) in the 48 h-static test and 15 microg/l (40% inhibition) in the 6 day-dynamic test, whereas 48 h-EC50 for daphnid mobility was 47 microg/l. The EC50s for lindane were 50 microg/l for daphnid grazing (48 h-static and 6 day-dynamic tests) and 383 microg/l for the immobilization test (48 h).     

Water quality and fish size affect toxicity of endosulfan, an organochlorine pesticide, to rainbow trout

The acute toxicity of endosulfan in juvenile rainbow trout (Oncorhynchus mykiss, 10.61+/-1.69 g) was evaluated in glass aquaria under static conditions. Nominal concentrations of endosulfan in the toxicity test ranged from 1.3 microg l(-1) to 29 microg l(-1). The concentrations of endosulfan that killed 50% of the rainbow trout within 24-h (24-h LC50), 48-h LC50, 72-h LC50, and 96-h LC50 were 19.78, 8.89, 5.28, and 1.75 microg l(-1), respectively. None of the unexposed control fish died, and the first fish died 4 h after exposure to 26.3 microg l(-1) of endosulfan. Survival of fish was significantly increased with increasing fish size and decreased with decreased fish size at the same temperature (p<0.001). Temperature also had a significant effect on survival of fish. Alkalinity at levels above 20 mg l(-1) as CaCO3 significantly increased survival of fish at 19.78 microg l(-1) of endosulfan. Increasing alkalinity from 20 mg l(-1) as CaCO3 to 42 or higher concentrations tested in this study (121 mg l(-1) as CaCO3) significantly increased survival of fish (p<0.01). Total hardness ranging from 55 mg l(-1) as CaCO3 to 126 mg l(-1) as CaCO3 did not affect survival of fish exposed to endosulfan. Endosulfan toxicity was found to be irreversible when fish were exposed to minimum concentrations of endosulfan tested. Histologically, fish gills had lamellar edema, separation of epithelium from lamellae, lamellar fusion, and swelling of the epithelial cells. Melanomacrophage centers were scattered throughout the trunk kidney, head kidney, and spleen. The liver of endosulfan-exposed fish had severe focal necrosis. None of these lesions were seen in unexposed control fish. Results indicate that alkalinity, temperature, and fish size affect endosulfan toxicity of rainbow trout.