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.     

In-situ partitioning of butyltin compounds in estuarine sediments

The in-situ solid/pore-water partitioning of tributyltin (TBT), dibutyltin (DBT) and monobutyltin (MBT) was determined for an estuarine sediment profile collected from a commercial marina. Total butyltin levels exceeded sediment quality guideline values, and were 220-8750 microg/kg for TBT, 150-5450 microg/kg for DBT and 130-4250 microg/kg for MBT. Pore-water butyltin concentrations ranged from 0.05 to 2.35 microg/l for TBT, 0.07-3.25 microg/l for DBT, and 0.05-0.53 microg/l for MBT. The partitioning of butyltin compounds between the sediment solid-phase and pore-water was described by an organic carbon normalised distribution ratio (D(OC)). The observed D(OC) values were similar for TBT, DBT and MBT, and were 10(5)-10(6) l/kg. Values for the Butyltin Degradation Index (BDI) were larger than 1 at depths greater than 10 cm below the sediment/water-column interface. This indicates that substantial TBT degradation has occurred in the sediments, and suggests that natural attenuation may be a viable sediment remediation strategy.     

Reproductive effects and bioconcentration of 4-nonylphenol in medaka fish (Oryzias latipes)

The objective of this study was to investigate the reproductive success (fertility and fecundity) of medaka (Oryzias latipes) exposed to 4-nonylphenol (4-NP) in water and the viability of their F(1) offspring. In addition, we measured the bioconcentration of 4-NP in eggs. After a 21-d exposure to 100microg/l 4-NP, medaka showed reduced egg production and fertility. Hepatic vitellogenin levels were increased significantly in males treated with 10, 50 and 100microg/l of 4-NP. In the F(1) generations, the hatchability and time to hatching of embryos in the 100microg/l treatment group were adversely affected, and 2-7microg 4-NP/g egg was found in spawned eggs (the bioconcentration factor: 30-100). These results indicate that (1) 4-NP adversely affected the reproduction (fecundity and fertility) of adult medaka, (2) 4-NP accumulated in eggs through maternal transfer and (3) these levels of 4-NP were associated with adverse effects in the F(1) offspring.     

Effects of tributyltin maternal and/or waterborne exposure on the embryonic development of the Manila clam, Ruditapes philippinarum

We examined the effect of tributyltin (TBT) on embryonic development of the Manila clam, Ruditapes philippinarum. In a maternal exposure test, 100 clams were exposed to TBT at measured concentrations of <0.01 (control), 0.061, 0.310, or 0.350 microg/l at 20-22 degrees C for 3 weeks, and the embryo developmental success (the ratio of normal D-larvae to all larvae) was measured. There was a significant negative correlation between embryo developmental success and TBT concentration in the female Manila clams (p < 0.001). These results indicated that TBT accumulated in the female clam decreased embryo developmental success. In a waterborne exposure test, fertilized eggs (4 h after fertilization) were exposed to TBT at measured concentrations of <0.01 (control), 0.062, 0.140, 0.320, or 0.640 microg/l for 23 h. Embryo developmental success was also significantly decreased in all TBT treatment groups compared with that in the control group. TBT accumulated in female adults and waterborne TBT clearly inhibit reproductive success of the clam.     

Effects of pharmaceuticals on Daphnia survival, growth, and reproduction

Pharmaceuticals have been globally detected in surface waters, and the ecological impacts of these biologically-active, ubiquitous chemicals are largely unknown. To evaluate the aquatic toxicity of individual pharmaceuticals and mixtures, we performed single species laboratory toxicity tests with Daphnia magna, a common freshwater zooplankton. We conducted acute (6-day) and chronic (30-day) exposure pharmaceutical bioassays and evaluated survivorship and morphology of adults and neonates, adult length, resting egg production, brood size (fecundity), and the proportion of male broods produced (sex ratio). In general, exposure to a single pharmaceutical in the 1-100 microg/l range yielded no apparent effects on the normal life processes of Daphnia. However, chronic fluoxetine exposure (36 microg/l) significantly increased Daphnia fecundity, and acute clofibric acid exposure (10 microg/l) significantly increased sex ratio. A mixture of fluoxetine (36 microg/l) and clofibric acid (100 microg/l) caused significant mortality; the same fluoxetine concentration mixed with 10 microg/l clofibric acid resulted in significant deformities, including malformed carapaces and swimming setae. Mixtures of three to five antibiotics (total antibiotic concentration 30-500 microg/l) elicited changes in Daphnia sex ratio. We conclude: (1) individual and mixtures of pharmaceuticals affect normal development and reproduction of Daphnia magna, (2) aquatic toxicity of pharmaceutical mixtures can be unpredictable and complex compared to individual pharmaceutical effects, and (3) timing and duration of pharmaceutical exposure influence aquatic toxicity.     

Chronic toxicity of tributyltin to development and reproduction of the European freshwater snail Lymnaea stagnalis (L.)

Chronic toxicity, growth and reproduction were measured in the freshwater gastropod Lymnaea stagnalis exposed to waterborne bis(tri-n-butyltin) oxide (TBTO) over a range of four nominal concentrations (0-10microg TBTl(-1)). Egg development was completely inhibited at 10microg TBTl(-1), whilst abnormal embryonic development was observed at 1microg TBTl(-1). For the solvent control and the 0.01microg TBTl(-1)treatment group, normal development of L. stagnalis was observed. Survivorship of hatchlings was significantly reduced by TBT at 1microgl(-1) while inhibition of shell growth of L. stagnalis was also observed at this concentration. The data were used to determine intrinsic growth rates (r) using two theoretical approaches (the Euler-Lotka equation and a Leslie Matrix). Both approaches showed that survival, fecundity and population growth rate were reduced at 1microg TBTl(-1). Interestingly, at 0.01microg TBTl(-1) snails showed a higher fecundity and growth rate than in the solvent control. The TBT concentration at which the r would equal zero (ECr(0)) and the population NOEC (No Observed Effect Concentration) were estimated. The population NOEC was defined as either the lower 95% confidence or lower 95% pointwise percentile limit of the ECr(0). Values obtained using the two different approaches were similar and thus a geometric mean was calculated to obtain a final representative population NOEC value for L. stagnalis of 2745ng TBTl(-1). The present data together with chronic toxicity TBT data for freshwater organisms, obtained from peer-reviewed literature, were used to construct a species sensitivity distribution (SSD). A predicted no effect concentration was then derived from the SSD (hazardous concentration at 5%, i.e., HC5 or 95% protection level). This SSD was compared with the SSD derived from saltwater species datasets. The HC5 value for saltwater species (3.55ng TBTl(-1); lower confidence limit: 1.93ng TBTl(-1)) was significantly lower than that for freshwater species (30.13ng TBTl(-1); lower confidence limit: 9.23ng TBTl(-1)), indicating that saltwater species are probably more susceptible to TBT than their freshwater counterparts.     

Acceleration of tributyltin chloride (TBT) degradation in liquid cultures of the filamentous fungus Cunninghamella elegans

In this study, we have examined the effects of synthetic medium ingredients and culture incubation conditions on growth and tributyltin chloride (TBT) degradation activity of the fungus Cunninghamella elegans. The best efficiency of TBT conversion to less toxic derivatives: dibutyltin and monobutyltin was noticed on media which contained glucose, NH(4)Cl, K(2)HPO(4) and MgSO(4). Next, the constructed M3 medium (with the above components) ensured vigorous growth of C. elegans and allowed the reduction of 80% of the initial TBT content (10 mg l(-1)), after 3d of biodegradation. The further acceleration of the biocide utilization by C. elegans was achieved by additional oxygen supply (pO(2) >or = 20%) to the growing fungus (89% after 2d of incubation in the BioFlo II bioreactor). The efficient xenobiotic biodegradation was related to the intensity of fungal growth. The obtained results suggest a cometabolic nature of TBT utilization by C. elegans.     

Effect of TBT on Ruditapes decussatus juveniles

The effects of sublethal concentrations of tributyltin (TBT) on growth of juvenile clams Ruditapes decussatus were determined during exposure to TBT concentrations of 50, 100 and 250 ng l(-1) (as Sn) for a period up to two years. Length and weight of clams increased continuously in all treatments throughout the experimental period, and, overall, rates were not significantly influenced by TBT exposure, although final length and weight were inversely related to increasing TBT concentration. Juvenile R. decussatus therefore appear to be less sensitive to TBT than larval stages. Some juveniles exposed to TBT developed abnormal shell growth, laterally, changing the typical flattened shape of clams into a more "rounded" form. This characteristic was more visible in the anterior margins of valves than posteriorly, and mainly observed in clams exposed to TBT at 50 ng l(-1) (as Sn).     

Fertilization success and sexual behavior in male medaka, Oryzias latipes, exposed to tributyltin

We evaluated the effects of tributyltin (TBT), polychlorinated biphenyls (PCBs), and a mixture of TBT and PCBs on reproduction, gonadal histology, and sexual behavior in Japanese medaka (Oryzias latipes). We administered TBT (1 microg g(-1) body weight [b.w.] daily), PCBs (1 microg g(-1) b.w. daily), or both to medaka for 3 weeks. We assessed reproductive success during week 3 and the sexual behavior of male medaka after the exposure period. Fertilization success was significantly decreased in groups that received TBT. The profiles of fertility developed a bimodal distribution in the TBT and TBT+PCBs groups, and the numbers of males that performed "following" and "dancing" were significantly decreased in the TBT+PCBs group. The frequency of "dancing" also decreased after treatment with TBT, whereas fish that received PCBs only showed no difference in sexual behavior. Testes that had oocytes showed normal spermatogenesis, even though two of the five males treated with TBT+PCBs had some indication of testis-ova, which did not occur in any other group. Our results indicate that TBT, but not PCBs, affects sexual behavior and reproduction in medaka.     

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.     

Contamination of butyltin and phenyltin compounds in the marine environment of Otsuchi Bay, Japan

Butyltin (BT) and phenyltin (PT) compounds were measured in seawater, sediment, and biological samples collected from coastal areas of Otsuchi Bay, Japan. Tributyltin (TBT) compounds in seawater, sediment, plankton, mussels, scallops and fish were in the range of 0.008-0.074 microg liter(-1), 0.01-0.64 mg (kg dry wt)(-1), 0.24-9.8 mg (kg dry wt)(-1), 0.04-0.18 mg (kg wet wt)(-1), 0.10-0.13 mg (kg wet wt)(-1) and 0.01-0.02 mg (kg wet wt)(-1), respectively. Trace amount of PTs were found in seawater. The highest concentrations of TBT and triphenyltin (TPT) were found near a shipyard. Triorganotin compounds were more dominant than their metabolites. A positive correlation was observed between the concentrations of TBT and TPT in the mussels. Concentrations of TBT and TPT in mussels were high in the upper intertidal zone, and decreased toward the water.     

Toxicity of fenvalerate to caddisfly larvae: chronic effects of 1- vs 10-h pulse-exposure with constant doses

Episodic pollution events such as runoff or spraydrift can lead to a short-term (few hours) contamination of aquatic ecosystems with pesticides. So far, different short-term exposures with respect to long-term effects have not been studied. In the present study, caddisfly larvae, typical for agricultural streams (Limnephilus lunatus Curtis, 2nd and 3rd instar) were exposed for 1- vs 10-h to three different equivalent doses (microg h) of fenvalerate. After transfer into an artificial stream microcosm with pesticide-free water, chronic effects were observed over 240 days. Comparison of 1- and 10-h exposure revealed that 1-h contamination leads to stronger effects. The differences were significant for the sublethal endpoints emergence pattern and dry weight of adults (ANOVA, Fisher's PLSD; P < 0.05). In terms of exposure dose, the difference between 1- and 10-h exposure equals a factor of 6 as a mean of all endpoints studied. The following significant effect levels for the 1-h exposure were obtained for the different endpoints investigated: reduced emergence success and production at 0.1 microg l(-1), temporal pattern of emergence at 0.001 microg l(-1), dry weight of adults at 0.01 microg l(-1).     

Comparison of heavy metal toxicity in life stages (spermiotoxicity, egg toxicity, embryotoxicity and larval toxicity) of Hydroides elegans

A toxicity test was developed to examine the effects of heavy metal contaminants on the early life stages of the marine polychaete. We have studied the effects of metals on fertilization and early development of marine polychaete Hydroides elegans. These heavy metals have often been found in polluted ground and water near industrial discharges, and have therefore been detected from time to time in the food chain. They have been reported to alter various reproduction functions in various animals including marine populations. The toxic effect of mercury, cadmium, lead, nickel and zinc on sperm viability, fertilization, embryogenesis and larvae of H. elegans was examined. We observed that the rate of fertilization decreased when the sperm was incubated with heavy metals. Treatment of eggs with each metal did not prevent fertilization, but delayed or blocked the first mitotic divisions, and altered early embryonic development. All these effects were observed at relatively high concentrations. However, bio-accumulation in sediments and aquatic organisms have been reported. Polychaete eggs may then be in contact with very high concentrations of these heavy metals in areas where these metals are not handled or stocked properly, and then develop into abnormal embryos. In addition to bivalves and sea-urchins, polychaete embryos can provide biological criteria for seawater quality standards taking into account the sensitivity of the invertebrates and their contribution in detection of harmful chemicals with no marked effect on the species. Our results indicate that the early development of H. elegans is highly sensitive to heavy metals and this polychaete can be routinely employed as a test organism for ecotoxicity bioassays in tropical and subtropical regions.     

Enhanced growth and reproduction of Caenorhabditis elegans (Nematoda) in the presence of 4-nonylphenol

The nematode Caenorhabditis elegans was exposed over a whole fife-cycle (72 h) to several concentrations of 4-nonylphenol (NP; nominal concentrations: 0-350 microg/l). Growth and reproduction of C. elegans were enhanced at NP concentrations of 66 and 40 microg/l, respectively, with effects showing dose-response relationships. These stimulatory effects might be of ecological relevance in benthic habitats, where organisms can be exposed to high concentrations of NP.     

Evaluation of municipal sewage treatment systems for pollutant removal efficiency by measuring levels of micropollutants

In order to evaluate the municipal sewage treatment systems used at Harbin municipal sewage treatment plant for their pollutant removal efficiency, raw sewage and effluent samples at different treatment stages from the sewage treatment systems were taken, priority pollutants (PPs) were identified and quantified using gas chromatography-mass spectrometry (GC-MS) and inductively coupled plasma-atomic emission spectrograph (ICP-AES). The test results indicated that there were one hundred and fifty species of organic pollutants identified in the raw sewage sample, and only ten species of PPs in all the sewage samples. The levels of dimethyl phthalate (DMP), diethyl phthalate (DEP), dibutyl phthalate (DBP), di-n-octyl phthalate (DnOP) in the sewage samples were 0.779-0.111 microg l(-1), 1.977-0.022 microg l(-1), 6.411-2.194 microg l(-1) and 7.152-2.953microg l(-1), respectively, and most of these phthalate esters (PAEs) were removed through anaerobic/aerobic (A/O) process; The levels of alachlor, acetochlor, atrazine were 0.074-0.021 microg l(-1), 0.160-0.096 microg l(-1) and 0.238-0.184 microg l(-1), respectively, and the total removal efficiency of atrazine was poorest through the sewage treatment systems. The levels of Cu, Cr, Se, Hg, Ni and Zn were 0.0030-0.2327 mg l(-1). It is therefore concluded from these results that the sewage treatment systems were efficient in removing most of the organic and inorganic compounds in this study, and so, the discharged effluent could cause little of the secondary pollution of the aquatic environment.     

Three decades of tributyltin in the coastal environment with emphasis on Arcachon Bay, France

This account briefly reviews the fate, bioconcentration and effects of tributyltin (TBT) three decades after its introduction into the coastal environment. Data on TBT-related events that occurred in Arcachon Bay (gastropod imposex, oyster recruitment failure, oyster shell malformations, concentrations in water and sediments) are revised and reinterpreted. Historically, TBT concentrations in the waters of the open bay seldom exceeded 1 ng Sn l(-1) in the 1960s, likely surpassed the range of 100 ng Sn l(-1) from 1977 to 1981 (years when no oyster spat were collected and the local shellfish industry collapsed), and decreased to about 1 ng Sn l(-1) in the late 1980s and early 1990s following legislation in January 1982 banning TBT use. TBT pollution is discussed in relation to the green tides that have developed in Arcachon Bay since 1982.     

Sensitivity of the zebrafish (Danio rerio) early life stage test for compounds with different modes of action

The sensitivity of the early life stage (ELS) toxicity test for two compounds with different modes of action was determined, and related to other toxicity tests with the same compounds. The zebrafish. Danio rerio, was used as a test organism, and the two model compounds were 1,2,3-trichlorobenzene (123TCB), a non-polar narcotic, and parathion, an acetylcholinesterase (AChE) inhibitor. Hatching and survival after 28 days were significantly reduced in the highest 123TCB treatment (263 microg/l), but not in any of the parathion treatments. Growth of the larvae was negatively affected at parathion concentrations above 20 microg/l, while AChE was only significantly inhibited at the highest concentration, 93 microg/l. No effects on growth were found in the 123TCB treatments. In comparison with acute and chronic studies with both compounds, the ELS test turned out to be less sensitive than chronic studies and more sensitive than acute studies. The difference in sensitivity between the tests systems seems however, to depend on the mode of action of the compound.     

Determination of butyltins in harbour sediment and water by aqueous phase ethylation GC-ICP-MS and hydride generation GC-AAS

Sediment and water samples have been collected between 1992 and 1995 for evaluating butyltin contamination in two marinas from the coastal area in The Netherlands, two years after the ban of TBT. Sediments extracts were analysed by capillary gas chromatography-inductively coupled plasma-mass spectrometry. Sediment concentrations for TBT showed no trend of decrease between 1992 and 1995 and were extremely high in the marina secluded from tidal action; 17.5 +/- 8.0 microg g(-1) and much lower for the marina with tidal action; 0.117 +/- 0.073 microg g(-1). High ratios between TBT and DBT in the sediments indicate that degradation mechanisms in the sediments are of minor importance. Dissolved butyltin compounds were analysed in water by means of gas chromatography-atomic absorption spectrometry. Water concentrations of TBT showed no clear trend of decrease between 1992 and 1995 and were high in the marina secluded from tidal action; 139 +/- 166 ng litre(-1) but much lower for the marina with tidal action; 48 +/- 98 ng litre(-1). An active degradation mechanism during summer periods was indicated by low (<1) TBT/DBT ratios in the water phase of the marina secluded from tidal action.     

Effects of Cadmium and Zinc on the feeding behaviour of two freshwater crustaceans: Atyaephyra desmarestii (Decapoda) and Echinogammarus meridionalis (Amphipoda)

Lethal and sublethal effects of Cadmium and Zinc on two freshwater crustaceans were investigated. The 96-h LC50 for Cadmium and Zinc were 42.20 microg l(-1) and 5.43 mg l(-1) for Atyaephyra desmarestii, and 36.17 microg l(-1) and 4.61 mg l(-1) for Echinogammarus meridionalis, respectively. An increase in metal concentrations at sublethal levels resulted in significant reductions of the feeding rate of both species. The LOECs in the chronic feeding assays for Zinc were 1.29 mg l(-1) for A. desmarestii and 0.4 mg l(-1) for E. meridionalis. For Cadmium the LOEC was 6.53 microgl(-1) for both species. The results suggest the potential use of these two crustacean species as test organisms in ecotoxicological tests. Feeding assays appear to be a rapid, cheap and effective tool to be used in biomonitoring studies of Portuguese freshwaters.     

Survival, growth and reproduction of Daphnia carinata (Crustacea: Cladocera) exposed to chronic cadmium stress at different food (Chlorella) levels

In nature, organisms have to respond to a diversity of factors acting simultaneously. The present investigation was conducted to study whether changes in food (Chlorella) levels could modify the chronic toxicity of cadmium on the various life-history parameters, such as survivorship, longevity, life expectancy, fecundity, age at first reproduction, R(0), T, r and growth rates of the cladoceran Daphnia carinata. The study indicated that at low food levels (0.5 x 10(6) cells ml(-1) Chlorella), cadmium concentrations in the range of 27-162 microg litre(-1) reduced these life-history parameters by 50% (EC(50)). At medium food levels (1.5 x 10(6) ml(-1) Chlorella) the EC(50) of cadmium was in the range of 51-127 microg litre(-1). At high food levels (4.5 x 10(6) cells ml(-1) Chlorella), the toxic effect of cadmium was greatly reduced. The decreases in survival, growth and reproduction of D. carinata at high cadmium-low food levels affected the fitness parameter 'r'. The study emphasises the need to include reproductive parameters other than mere survival in toxicity bioassays. The study also stresses the need to incorporate in laboratory tests other relevant factors that might modify pollutant toxicity.