Toxicity of copper to three estuarine bivalves

This paper reports on a study of the toxicity of copper to three species of bivalve of commercial food value, Anadara granosa (Linnaeus), Meretrix casta, Deshayes and Crassostrea madrasensis (Preston) inhibiting the Vellar estuary at Porto Novo (southern India). The LC₅₀ values were estimated as 60 μg Cu/litre for A. granosa, 72 μg Cu/litre for M. casta and 88 μg Cu/litre for C. madrasensis subjected to 96-h static bioassay tests after acclimation for a period of four days in the laboratory. The animals selected for study ranged in length from 29 to 44 mm in A. granosa, from 25 to 42 mm in M. casta and from 31 to 115 mm in C. madrasensis. The experiments were conducted at 25% salinity with pH 8·0 ± 0·1 at a temperature of 27°C ± 0·5°C. The revival rates of bivalves which survived exposure to the LC₅₀ concentrations for the 96-h period (when marked and released in the natural environment) were 67% for A. granosa, and C. madrasensis and 83% for M. casta. The LT₅₀ values for the three bivalves at different concentrations of the metal were also calculated.     

Effects of long-term exposure to silver or copper on growth, bioaccumulation and histopathology in the blue mussel Mytilus edulis

The purpose of this study was to determine the long-term accumulation of either silver or copper from low concentrations in seawater by blue mussels, Mytilus edulis. Mussels raised from eggs in the laboratory to the age of 2·5 months (approximately 4·5 mm in length) were continuously exposed to 0, 1, 5 and 10μg/liter of either silver (nitrate) or copper (chloride) and sampled at 12, 18 and 21 months for growth studies, measurements of metal accumulation and histopathological examination.Whole-body soft tissues were analyzed for the presence of both silver and copper, as background levels of copper in the incoming seawater averaged 2–4 μg/liter. Mussels exposed to silver had accumulated significant amounts of silver only at the highest test concentration (10 μg/liter Ag) after 12 months, but at 18 and 21 months significant levels were accumulated at all three test concentrations. Mussels exposed to copper accumulated significant amounts of copper at 5 and 10 μg/liter Cu after all three sampling periods, but not at 1μg/liter. Silver-exposed animals also accumulated significantly greater amounts of copper than control animals.In a comparative study, field-collected juvenile mussels (approximately 16·1 mm in shell length) and adult mussels (approximately 53·4 mm in shell length) were exposed for 12 months to 0, 5, 25 and 50 μg/liter silver only and subsequently sampled for metal-accumulation analyses and growth measurements. Juvenile mussels accumulated significant amounts of silver at all test concentrations, with the exception of mussels exposed to 5 μg/liter Ag for 6 months. Copper accumulation in the silver-exposed juveniles was significant only at 50 μg/liter Ag after 6 months, but at all test concentrations after 12 months. Adult mussels exposed to silver accumulated significant levels of both silver and copper, but at somewhat lower levels than juveniles.In the growth study, silver had no effect on laboratory reared mussels at the highest concentration of 10 μg/liter tested, whereas copper at 10 μg/liter did appear to affect growth as early as 4 months after the start of experimental exposure. Field-collected juvenile mussels did show inhibition in growth after 6 months' exposure to 25 and 50 μg/liter Ag, with some growth occurring after 12 months. Adults also showed inhibition in growth after 6 months but not at 12 months.Histopathological examination of mussels exposed to either 5 or 10 μg/liter of copper for 18 months showed changes in the digestive diverticula, gastrointestinal tract, reproductive tract and muscle tissues. These changes were more noticeable in mussels exposed to 5 μg/liter Cu than in those exposed to 10 μg/liter. Mussels exposed to silver for 21 months showed yellowish to black particulate deposition in the basement membrane and connective tissue of the various organs and tissues. Silver deposition increased with increasing test concentration.     

Organic complexation and its control of the dissolved concentrations of copper and zinc in the Scheldt estuary

Cathodic stripping voltammetry (CSV) is used to determine total (after UV-irradiation) and labile dissolved metal concentrations as well as complexing ligand concentrations in samples from the river Scheldt estuary. It was found that even at high added concentrations of catechol (1 m for copper and 0·4 m for iron) and of APDC (1 m for zinc) only part of the dissolved metal was labile (5–58% for copper, 34–69% for zinc, 10–38% for iron); this discrepancy could be explained by the low solubility of iron which is largely present as colloidal material, and by competition for dissolved copper and zinc by organic complexing ligands. Ligand concentrations varied between 28 and 206 n for copper and between 22 and 220 n for zinc; part of the copper complexing ligands could be sub-divided into strong complexing sites with concentrations between 23 and 121 n and weaker sites with concentrations between 44 and 131 n . Values for conditional stability constants varied between (logK′ values) 13·0 and 14·8 for strong and between 11·5 and 12·1 for weaker copper complexing ligands, whereas for zinc the values were between 8·6 and 10·6. The average products of ligand concentrations and conditional stability constants (a-coefficients) were 6 × 10² for zinc and 6 × 10⁶ for copper.The dissolved zinc concentration was found to co-vary with the zinc complexing ligand concentration throughout the estuary. It is argued that the zinc concentration is regulated, in this estuary at least, by interactions with dissolved organic complexing ligands. A similar relationship was apparent between the dissolved copper and the strong copper complexing ligand concentration. The total copper complexing ligand concentrations were much greater than the dissolved copper concentrations, suggesting that only strongly complexed copper is kept in solution.These results provide evidence for the first time that interactions of copper and zinc with dissolved organic complexing ligands determine the geochemical pathway of these metals.     

Evidence on the interaction of mercury and selenium in the shrimp Palaemon elegans

The interaction of selenium with mercury was studied in the shrimp Palaemon elegans. The release of ²⁰³HgCl₂ (5·0 μg Hg per gramme body weight) from shrimp pretreated with selenium (SeO₂ doses of 1·97, 3·95 and 7·90 μg Se per gramme fresh weight) was significantly decreased compared with the control group to which only 5·0 μg Hg/g had been administered.In the presence of HgCl₂ (5·0 μg Hg per gramme fresh weight) the release of ⁷⁵Se also diminished significantly at the higher stable Se pretreated dose (7·90 μg Se/g) while, at a lower selenium concentration, the release was not statistically different. Analyses for stable Hg and Se confirmed the decrease in rate of selenium loss in the presence of mercury which had been demonstrated with radiotracers.A dose of 7·9 μg Se per gramme fresh weight injected 12 h before exposure of the shrimp to the various mercuric chloride solutions did not produce a significant difference in the 24 h LC₅₀ compared with the group not pretreated with selenium. However, during exposure to mercury at 3·8 mg/litre, the median lethal time (LT₅₀) for the shrimp pretreated for 4 days with sublethal selenium (6·9 and 10·5 mg Se/litre) was delayed (19·2 and 33·2 h) compared with the group which was not pretreated. The results are discussed in relation to the role of selenium in the acutely toxic effects of inorganic mercury.     

Fate of tributyltin in sewage sludge treatment

We have shown that municipal wastewater and sewage sludge are contaminated by organotins. Here, we investigate the behaviour and fate of tributyltin (TBT) in laboratory-scale sewage sludge treatment at various conditions including both aerobic and anaerobic, and mesophilic and thermophilic. Organotins were determined by capillary GC-FPD. Fresh raw sewage sludge was stored in a tank that was connected to five chemostats maintained at different conditions. Raw sludge contained TBT, dibutyltin (DBT) and monobutyltin (MBT) residues in the range of 0·28 to 0·83 mg/kg (dry weight). The volumetric TBT concentration was 10·5 μg/litre in the influent, and ranged from 7·4 to 8·3 μg/litre in the effluent of the different chemostats. In the anaerobic mesophilic treatment, concentrations of TBT did not decrease with increasing sludge residence time, DBT increased and MBT showed no clear trend. The DBT increase is suggested to be due to leaching from the PVC walls of the chemostats, and not to degradation of TBT alone. The TBT reduction did not show significant differences between the mesophilic and thermophilic anaerobic fermentors. These findings indicate that the degradation of TBT during sludge treatment is only low under all conditions tested.     

Sodium hexametaphosphate as an aid in benthic sample sorting

The efficacy of sodium hexametaphosphate, when added to a formalin-Rose Bengal fixative solution, in improving the efficiency of benthic sample sorting by encouraging the disaggregation of silt-clay sediment has been investigated. Samples of estuarine mud (Mdφ = 6·1) from Damhead Creek, River Medway, were used for the analysis. A significant increase (23%; p < 0·01) in sieving efficiency was found in samples treated with concentrations of 1·24g/litre or more of sodium hexametaphosphate; treatments with higher concentrations than 1·24g/litre did not improve on this performance significantly. The use of 1·24g/litre sodium hexametaphosphate in an initial fixative/preservative solution is recommended as an economic and practical means of improving the efficiency of benthic sample sorting, without deleterious effects on the animal specimens in the sample.     

Reproductive success, xenobiotic contaminants and hepatic mixed-function oxidase (MFO) activity in Platichthys stellatus populations from San Francisco Bay

To identify some specific effects of organic contaminants on fisheries in an urbanized estuary we compared the reproductive success of starry flounder from San Francisco Bay with concentrations of tissue contaminants and hepatic mixed-function oxidase (MFO) activity. We found significantly lower (P < 0·05) sediment concentrations of total identified polynuclear aromatic hydrocarbons (PAHs) in the less urbanized San Pablo Bay (SP) area (Fig. 1) than in the more urbanized central bay (CB) stations (Table 1). For flounder in early gametogenesis (August and September) the SP fish (n = 20) had significantly lower (P < 0·01) liver concentrations of Aroclor 1260 (0·34 ± 0·14 μg/g) than those at the CB stations: Berkeley (BK, n = 20, 1·6 ± 1·6 μg/g); Oakland (OK, n = 16, 2·3 ± 2·8 μg/g); and Alameda (AL, n = 4, 2·2 ± 1 μg/g). A similar pattern existed for DDT concentrations: SP = 0·2 ± 0·16 μg/g; BK = 0·1 ± 0·34 μg/g; OK = 0·4 ± 0·53 μg/g; and AL = 0·4 ± 0·33 μg/g. Total PAHs in livers were as follows: SP = 0·14 μg/g; BK = 2·6 μg/g; OK = 1·4 μg/g; and AL = 14 μg/g. Although gonad index, liver index, and presence of fin rot are inversely related to aryl hydrocarbon hydroxylase (AHH) activity, healthy fish in a similar reproductive state have lower AHH activities in the SP area. For example, in August and September, 1984, mean AHH activities were as follows: SP = 203 ± 89, and CB = 355 ± 200 pmol 3-OH-B[a]P mg microsomal protein min. We found a log-linear relationship for AHH activity and its percent inhibition by 7,8-benzoflavone (10⁻⁴m) and only a few fish from SP showed enhanced AHH activity after addition of 7,8-benzoflavone. This suggests that most of the starry flounder in San Francisco Bay are induced.     

Trace metal fronts in Scottish coastal waters

Dissolved cadmium and copper concentrations have been determined in 76 surface water samples in coastal and ocean waters around Scotland by anodic stripping voltammetry (ASV). A trace metal/salinity ‘front’ is observed to the west, north and north-east of Scotland separating high salinity ocean water (>35 × 10⁻³) with low concentrations of dissolved Cd and Cu from lower salinity (<35 × 10⁻³) coastal water containing higher concentrations of Cd and Cu. Mean Cd concentrations in ocean and coastal waters are 7 ng dm⁻³ (0·06 n ) and 11 ng dm⁻³ (0·10 n ) respectively; for Cu the respective levels are 60 ng dm⁻³ (0·95 n ) and 170 ng dm⁻³ (2·68 n ). The observed distribution is attributed principally to freshwater runoff and the advection of contaminated Irish Sea water into the study area.     

Use of an ion exchange technique to measure copper complexing capacity on the continental shelf of the southeastern United States and in the Sargasso Sea

An ion exchange technique has been used to determine the copper complexing capacity (CuCC) of strong organic complexing agents at 21 stations across the continental shelf of the southeastern United States and in the western Sargasso Sea. The concentration of dissolved organic carbon (DOC) and total particulate materal (TPM), two pools of potential complexing agents, was also measured at each station. The CuCC ranged from 0.014 to 1.681 μM Cu dm⁻³ on the inner shelf, from 0.043 to 0.095 μM Cu dm⁻³ in mid and outer shelf waters, and from < 0.010 to 0.036 μM Cu dm⁻³ at the Sargasso Sea stations. The correlation between CuCC and both DOC and TPM is highly significant (α < 0.01). Two synoptic surveys of the distribution of DOC and TPM across the shelf showed that DOC ranges from > 3 mg C dm⁻³ nearshore to <1 mg C dm⁻³ offshore and that TPM ranges from > 50 mg dm⁻³ nearshore to <1 mg dm⁻³ offshore. Both TPM and DOC are most variable on the inner shelf. These data are consistent with CuCC data which indicate that the CuCC of inner shelf waters was relatively high and very heterogeneous. In contrast, DOC, TPM and copper complexing capacity are low and nearly invariant at the Sargasso Sea stations. We present a model of the distribution of complexing agents in different marine environments and hypothesize that the mechanisms underlying differences between environments relate to differences in the source(s) and nature of complexing agents in each system.     

The combined effect of copper and hydrocarbons on the length growth of Mytilus edulis

The combined effect of copper and North Sea crude oil is tested on juvenile Mytilus edulis. Dissolved CuSO₄ x 5H₂O is added to the seawater in concentrations of 3 and 6 μg Cu litre^-1. Crude oil, microencapsulated in a gelatine/acacia structure, is added in concentrations of 1·5 and 3·0 mg oil litre^-1. The shell length growth is measured every 24 h with laser diffraction. Linear effects were observed for single components within 24 h of exposure and significant antagonistic interaction appeared within 48 h. Reduced growth rate coincided with reduced filtering and defaecation, and reduced byssus production.     

Long-term effects of tributyltin compounds on the baltic amphipod, Gammarus oceanicus

Amphipod larvae, Gammarus oceanicus, were exposed for 8 weeks to bis(tributyltin) oxide (TBTO) or tributyltin fluoride (TBTF), or for 5 weeks to leachates from tributyltin-containing antibiofouling paints. No larvae survived 8 weeks' exposure to 3 μg/litre TBTO or TBTF. Differential larval survival occurred during exposure to 0·3 gmg/litre of these same compounds. Paint leachates were shown to cause similar toxicity responses at comparable aqueous tributyltin concentrations. An examination of sublethal responses showed slight decreases in growth for larvae exposed to TBTO and to paint leachates, although responses at tributyltin levels below 1 μg/litre were not marked. Whole-animal oxygen consumption rates were not affected by low tributyltin exposure levels. While these studies demonstrate low level toxicity of tributyltins, effects on oxygen consumption and growth do not indicate a mode of action for these compounds.     

Chlorination by-products in chlorinated cooling water of some European coastal power stations

Chlorination by-products (CBPs) are formed as a result of the cnlorination of power station cooling water for anti-fouling purposes. Their production was studied at 10 coastal power stations in the UK, France and The Netherlands. Three categories of CBPs were determined: trihalomethanes; haloacetonitriles; and halophenols. Bromoform was the CBP most abundantly present in the effluents of all 10 power stations. At a mean chlorine dosage of 0.5–1.5 mg/litre (as Cl₂) the mean bromoform concentration was 16.32 ± 2.10 μg/litre. The CBP found in second highest concentrations was dibromoacetonitrile (DBAN) with mean concentrations of 1.48 ± 0.56 μg/litre. Other CBPs detected were dibromochloromethane, bromodichloromethane and 2,4,6-trichlorophenol; concentrations of these three compounds were very low (< 1 μg/litre). At those sites at which bromoform was measured in the dispersing effluent plume it was found to behave as a conservative parameter (Significant direct correlation with plume ΔT).     

Acute and sublethal toxicity of naphthalene and three methylated derivatives to the estuarine copepod, Eurytemora affinis

The bi-cyclic aromatic hydrocarbon naphthalene (N) and its alkylated derivatives 2-methylnaphthalene (MN), 2,6-dimethylnaphthalene (DMN) and 2,3,5-trimethylnaphthalene (TMN) were used to investigate possible relationships between increased methylation and toxicity during acute high level and chronic low level exposure experiments using the calanoid copepod Eurytemora affinis Poppe. The 24-h LC₅₀ levels determined indicated an approximate twofold increase in toxicity per methyl group addition: N-3798 μg/litre; MN-1499 μg/litre; DMN-852 μg/litre and TMN-316 μg/litre. Low level chronic exposure of E. affinis to the various naphthalenes in sea water concentrations of about 10 μg/litre for the duration of their adult life (maximum twenty-nine days) resulted in significant reductions in length of life, total numbers of nauplii produced and mean brood size. Exposure to all the naphthalenes at this level (10 μg/litre) resulted in reduced rates of egg production which were, on average, about 50% of those of control animals. Results are discussed in relation to previous work on the fate and effect of petroleum hydrocarbons in marine copepods.     

Contrasting complexing capacity of dissolved organic matter produced during the onset,development and decay of a simulated bloom of the marine diatom Skeletonema costatum

The capacity of natural dissolved organic matter (DOM) produced during the onset, development and decay of a simulated bloom of the marine diatom Skeletonema costatum to complex free copper has been followed for a 2 week period. Copper binding capacity of the culture was measured by anodic stripping voltammetry (ASV) with a hanging mercury drop electrode (HMDE). The concentration of dissolved organic carbon (DOC) and two fluorophores, M (humic-like, Ex/Em: 320 nm/410 nm) and T (protein-like, Ex/Em: 280 nm/350 nm), were followed during the course of the incubation. Models using DOC concentrations alone could not accurately predict the complexing capacity of the culture, especially at the end of the bloom, and better predictions were obtained when fluorescence measurements were considered. They were helpful in characterising two types of copper ligands produced in the culture. The first type, traced by the fluorescence of peak T, was related to labile DOC directly exuded by phytoplankton. The second type, traced by the fluorescence of peak M, was the refractory humic-like material presumably produced in situ as a by-product of the bacterial degradation of phytogenic materials. During the onset and development of the bloom (days 0 to 7), the fluorescence of peak T explains 60–80% of the total complexing capacity of the culture, suggesting that exuded “protein-like” compounds among other exuded complexing agents efficiently complexed free copper. On the contrary, during the decay (days 8 to 13), these ligands were replaced by humic substances as the complexing agent for copper.     

Adaptation of the polychaete Neanthes Arenaceodentata to copper

Neanthes arenaceodentata were exposed to 292, 146, 92 and 56 μg litre⁻¹ Cu (measured) and control seawater after a 27-day pre-exposure to a sublethal concentration of Cu (10, 16 and 28 μg litre⁻¹ and control) to determine if the worms increased their tolerance to Cu after the pre-treatment. The worms pre-exposed to 28 μg litre⁻¹ Cu were significantly more resistant to Cu toxicity than control and 10 and 16 μg litre⁻¹ Cu pre-exposed worms. For example, the time to 50 % mortality at 92 μg litre⁻¹ Cu was 18 and 11 days for worms pre-exposed to 28 μg litre⁻¹ Cu and control conditions, respectively. The net rate of Cu uptake during the toxicity test was lower for worms pre-exposed to 28 μg litre⁻¹ Cu than for the control and 10 and 16 μg litre⁻¹ Cu pre-exposed worms. For example, the net rate of Cu uptake at 292 μg litre⁻¹ Cu by worms pre-exposed to 28 μg litre⁻¹ Cu and control conditions was 42 and 102 μg g⁻¹ day ⁻¹, respectively.     

Copper complexation by fulvic acid affects copper toxicity to the larvae of the polychaete Hydroides elegans

Copper toxicity is influenced by a variety of environmental factors including dissolved organic matter (DOM). We examined the complexation of copper by fulvic acid (FA), one of the major components of DOM, by measuring the decline in labile copper by anodic stripping voltammetrically (ASV). The data were described using a one-site ligand binding model, with a ligand concentration of 0.19 μmol site mg⁻¹ C, and a log K′ of 6.2. The model was used to predict labile copper concentration in a bioassay designed to quantify the extent to which Cu–FA complexation affected copper toxicity to the larvae of marine polychaete Hydroides elegans. The toxicity data, when expressed as labile copper concentration causing abnormal development, were independent of FA concentration and could be modeled as a logistic function, with a 48-h EC₅₀ of 58.9 μg l⁻¹. However, when the data were expressed as a function of total copper concentration, the toxicity was dependent on FA concentration, with a 48-h EC₅₀ ranging from 55.6 μg l⁻¹ in the no-FA control to 137.4 μg l⁻¹ in the 20 mg l⁻¹ FA treatment. Thus, FA was protective against copper toxicity to the larvae, and such an effect was caused by the reduction in labile copper due to Cu–FA complexation. Our results demonstrate the potential of ASV as a useful tool for predicting metal toxicity to the larvae in coastal environment where DOM plays an important role in complexing metal ions.     

Cadmium, copper, lead and zinc in three species of planktonic crustaceans from the east coast of Corsica

Results of trace metal analyses performed on two species of Euphausiacea, Meganyctiphanes norvegica and Stylocheiron longicorne, and one species of Decapoda, Sergestes arcticus, collected off the east coast of Corsica, are reported. Analyses were carried out by atomic absorption spectrophotometry and by differential pulse anodic stripping voltammetry.S. arcticus contained lower concentrations of phosphorus (which was also analysed as a biological indicator), cadmium (0.33 μg g⁻¹), copper (17.7 μg g⁻¹), lead (2.13 μg g⁻¹) and zinc (51 μg g⁻¹) than the two Euphausiacea (0.50 μg Cd g⁻¹, 25.4 μg Cu g⁻¹, 4.03 μg Pb g⁻¹ and 59 μg Zn g⁻¹). Moreover, manganese concentrations were low in all the samples.When the results presented here are compared with previous results on phytoplankton and mesozooplankton, there appears to be no trend of trace metal enrichment from phytoplankton to the Decapoda.     

The behaviour of a Cu(II) ion selective electrode in seawater; copper consumption capacity and copper determinations

Determinations of copper consumption capacity (CuCs.C) and labile copper concentrations in surface coastal seawater, using a copper ion selective electrode (Cu-ISE) potentiometric method under predominantly diffusive conditions, are reported. For evaluation of the copper concentrations, the points of the endpoint contiguity zone of the CuCs.C titration curve were treated by an ISE multiple standard addition technique. The results were compared with those obtained by means of a Chelex-100 (calcic form) ‘batch’ procedure-potentiometric stripping analysis.The labile copper of the sample was determined at concentrations down to 10.70 nM with an average RSD of 12%, independent of the Cu-ISE employed. For adjacent subsamples, the mean CuCs.C values obtained for El Way seawater were equivalent to 81.05 and 48.00 nM copper, with an RSD of 4 and 7%, and for Isla Santa Maria seawater the value was equivalent to 70.27 nM copper, with an RSD of 7%. The theoretical approach of the electrode diffusive mechanism proposed, which would depend, fundamentally, on the adsorptive, complexing and reducing properties of the dissolved organic matter in the seawater sample, allows simultaneous analytical determination of CuCs.C and labile copper concentration in seawater.     

The distribution of total and electrochemically available copper in the northwestern atlantic ocean

Copper in a series of samples from the northwestern Atlantic has been determined by anodic stripping voltammetry (ASV) and atomic absorption spectrometry after preconcentration by cobalt—APDC coprecipitation. Samples from a transect across the continental shelf directly off the New York Bight showed a linear relationship between total copper and salinity (31–34‰) and less than 5% of the total copper was detectable by ASV. A slope water station showed less than 9% of the total copper to be ASV detectable in the upper 780m, below which the ASV detectable copper increased to 70% at 3000 m. The Sargasso Sea profiles showed less than 7% ASV detectable copper in the upper 150 m; the fraction of the total copper detected by ASV increased to about 80% of the total copper at greater depths. The reduced availability of copper to the ASV measurement in coastal waters and open ocean surface waters appears to be related to recent contact with waters where biological productivity occurs.     

Some effects of certain metals on development and mortality within the moult cycle of crangon crangon (L.)

Crangon crangon (L.) (Crustacea: Decapoda) were subjected to three metals (Cd, Cu and Zn) at respective concentrations (0·35, 1·6 and 14·4mg/litre) that would induce 50% mortality in 5–6 days. Significantly different (P < 0·05) rates of mortality occurred in the three groups, although their ET₅₀'s were almost identical. No moultstage-dependent mortality was observed for Cd, but in Cu and Zn the post-moult stages (A and B) were more susceptible than either the intermoult (C_a and C_b) or premoult (D₀–D₄) stages.No significant (P> 0·05) sex- or size-dependent mortality was evident at the tested concentrations of these metals.Feeding and ecdysis were inhibited in all metals after day 3 and development from one moult-stage to the next was greatly reduced in Cd.Possible modes of toxicity have been discussed in terms of the physiological condition inferred from the moult-stage at the time of death.