Effects of copper and zinc on two planktonic ciliates

The interactive effects of copper and zinc on two estuarine planktonic ciliates, Favella sp. and Balanion sp., were determined in seawater media in which the free metal ion activities were controlled by nitrilotriacetic acid (NTA) trace metal ion buffer systems. Cupric ion activities of 10^-10 M caused abnormal motility in both ciliates in shortterm (5 h) tests, and cupric ion activities as low as 10^-12.8 M decreased the growth rates of both species in longer-term experiments. In the short-term tests, zinc ion activity by itself did not affect the motility of the ciliates, but there were significant interactions between copper and zinc. In the longer-term experiments, the growth of Favella sp. was optimal at the lowest cupric ion activity (10^-13 M) and the two lowest zinc ion activities (10^-12 and 10^-13 M) and the two lowest zinc ion activities (10^-12 and 10^-11 M), and copper and zinc inhibited growth at activities above these values. By contrast, optimal growth rate of Balanion sp. occurred at the highest zinc ion activity (10^-10 M) and the lowest cupric ion activities (10^-13 to 10^-12 M) and growth rate was reduced at zinc ion activities 10^-11 M. There was an antagonism between copper and zinc which was particularly pronounced in Balanion sp.Contribution No. 5871 from the Woods Hole Oceanographic Institution     

Effects of cupric and zinc ion activities on the survival and reproduction of marine copepods

The toxicity of copper and zinc to the estuarine copepod Acartia tonsa and to the two diatom food species Thalassiosira pseudonana and T. weissflogii was measured in nitrilotriacetate-trace metal ion buffer systems at 25 S. Overall, A. tonsa appeared to be more sensitive to cupric and zinc ion activity than either of the diatoms; however, its sensitivity varied among the different life stages examined. Adult survival was not affected within the zinc ion activity range 10^-11 to 10^-8 M and cupric ion activity range 10^-13 to 10^-11 M over a 96-h period, but a cupric ion activity of 10^-10 M caused total mortality of adults within 72 h. Egg-laying rate was most sensitive to zinc, and was reduced at zinc ion activities 10^-10 M. Naupliar survival after 96 h was reduced by zinc ion activities 10^-8 M and by cupric ion activities 10^-11 M, and was reduced to zero at a zinc ion activity of 10^-7 M and at a cupric ion activity of 10^-10.5 M. In an interspecies comparison of 96-h adult survival, Centropages typicus was more sensitive to copper and zinc than A. tonsa and the survival of Labidocera aestiva was dependent on the ratio of cupric to zinc ion activity. A comparison of our results with estimates of zinc and cupric ion activities in estuaries suggests that ionic activities of these metals are high enough in some polluted estuaries to affect the survival and reproduction of copepods.     

Combined effect of the chlorides of mercury and copper in sea water on the euryhaline amphipod Gammarus duebeni

The possible interactive effect of the chlorides of copper and mercury on the euryhaline amphipod Gammarus duebeni in 100% sea water was examined using the following indices: (i) 96 h LC50 values, (ii) urine production rates and (iii) degree of mercury accumulation. Both (a) the interaction of the chlorides of mercury and copper together in solution and (b) the influence of cupric chloride pre-treatment of individuals prior to exposure to mercuric chloride were investigated. Presence of a sublethal level of cupric chloride protected G. duebeni against the toxic action of mercuric chloride. Cupric chloride pretreatment was not so effective. The nature of the interaction between mercury and copper is discussed.     

Toxicity of cupric lon to eggs of the spot Leiostomus xanthurus and the Atlantic silverside Menidia menidia

Experiments were conducted to determine the toxicity of free cupric ion to the eggs of two marine fish: the spot Leiostomus xanthurus, and the Atlantic silverside Menidia menidia. A cupric ion buffer consisting of 5 mM of the chelator, trishydroxymethylamino methane (Tris), and varied concentrations of CuSO₄ was employed to achieve stable cupric ion activities in experimental seawater media. Egg hatch of the silverside appears to be more sensitive to cupric ion than that of the spot. The silverside had 60% inhibition of hatch at a pCu (negative logarithum of cupric ion activity) of 9.4 with complete supporession of hatching at pCu values below 8.2, while the spot had 50% inhibition at a pCu of 9.0 and complete suppression at a pCu of 8.0. Cupric ion was most toxic to the eggs of silverside at or around the time of hatching, whereas a pronounced sensitivity at hatching was not apparent with the eggs of spot. Calculated estimates of the range of cupric ion activity in seawater indicate that natural activity levels may in some instances inhibit the hatching of silverside and spot eggs.     

Physiological responses ofCalanus finmarchicus andMetridia longa (Copepoda: Calanoida) during the winter-spring transition

The effect of pH on the biological availability of copper to the brine shrimp Artemia franciscana was studied with acclimated and non-acclimated individuals for the pH range 5.5 to 8.5. A chemical speciation model was used to calculate the speciation of copper in a chemically-defined saline solution as a function of pH. The lipid solubility of inorganic copper species was determined in hexadecane/saline and octanol/saline extraction systems. Copper is absorbed across the gut epithelium and accumulation is proportional to time over a 120 min experimental period. The biological availability of copper decreases with a decrease in the pH of the salt solution and a concomitant increase of the cupric ion concentration. Acclimation to the experimental pH has a marked effect on the uptake process, which depends on the buffer used. There is no extraction of copper in hexadecane, but a small amount goes in octanol. There is, however, no direct relation between the accumulation of copper in the shrimps and the extraction of copper in octanol. Multiple regression of the accumulation rates for non-acclimated animals on the calculated copper species concentrations shows that much of the variation in accumulation rate with pH is explained when copper hydroxide and/or copper carbonate species are considered to be the biologically available forms. Alternatively, the observations can be interpreted as the result of competitive binding of protons and copper species for carrier systems.     

Relative contributions of food and water in the accumulation of zinc by two species of marine fish

A model food chain, utilizing ⁶⁵Zn-labeled and nonlabeled food organisms, was used to measure the relative contributions of food and water to Zn accumulation by Gambusia affinis and Leiostomus xanthurus. Chlamydomonas sp. was fed to Artemia sp. which in turn was fed to G. affinis and L. xanthurus. A trace metal-chelate buffer system was used to maintain a stable free Zn ion activity (10^-8.5 mol l^-1) in the experimental seawater. Food represented 78 to 82% of total accumulation of ⁶⁵Zn by the fish. Thus, food cannot be ignored in assessing the accumulation and toxicity of trace metals.     

Metal interaction during accumulation by the mussel Mytilus edulis planulatus

Mytilus edulis planulatus (Lamarck) were collected from Howden, South-east Tasmania in autumn 1981. Interaction effects of cadmium, copper and zinc during accumulation by mussels exposed for ten days to all three metals simultaneously were examined in a series of experiments in which each metal was tested at three concentrations. In general, interaction effects were most evident at the highest concentrations tested (20 g l^-1 Cd; 20 g l^-1 Cu; 200 g l^-1 Zn) and led to a reduction in the accumulation of cadmium and an increase in that of copper and zinc. More specifically, high levels of zinc caused a decrease in cadmium uptake and an increase in copper accumulation. The presence of copper resulted in depressed cadmium accumulation while zinc accumulation increased. Cadmium tended to enhance zinc accumulation, but copper accumulation was only affected to any great extent when zinc was also present.     

Effects of calcium,strontium, and magnesium on the coccolithophorid Cricosphaera (Hymenomonas) carterae. II. Cell division

The effects of calcium-deficiency, magnesium-deficiency and strontium enrichment on cell division in the alga Cricosphaera (Hymenomonas) carterae have been studied. Cell growth was reduced at 10^-3 M Ca and was absent at 5x10^-4 M Ca and lower concentrations. The addition of Sr to Ca-deficient media enabled cells to divide, the effect increasing with Sr concentration. When 4.6x10^-4 M Sr was added to media containing 10^-4 M Ca, the rate of division and the final cell concentration were comparable to the control (10^-2 M Ca). Strontium was 20 times more effective than Ca. The rate of growth was also examined at various Mg concentrations. Cell division was absent, or nearly so, in Mg concentrations below 4.2x10^-5 M. Cell size increased progressively as the Mg concentration decreased; however, the lowest protein concentration was found in the absence of Mg. In media lacking Mg, cells exhibited changes in ultrastructure including rounding-up and apparent fragmentation of chloroplasts and an increase in vacuole size. Also, the number of mitochondria per cell section increased 2.9 times in the absence of Mg while the total cross-sectional area remained the same, indicating fragmentation.     

Combined effects of copper and cadmium on Chlorella pyrenoidosa H.Chick: subcellular accumulation,distribution, and growth inhibition

Disposal of waste into aquatic ecosystems may cause microalgae to be exposed to various metals, e.g. copper and cadmium. The effects caused by combinations of metals may be more serious. Evaluations of subcellular fate, bioaccumulation, and biological effects of metals on aquatic organisms are generally derived from experiments with individual metals. The present study aims to evaluate the effects of exposure of Chlorella pyrenoidosa to copper and cadmium in combination on subcellular accumulation, distribution, and growth. The algae were exposed for 72 h to copper at concentrations of 13 – 25 µmol L^?1, cadmium at about 6 µmol L^?1, and combinations thereof. The levels of copper and cadmium in subcellular organelles, heat-denaturated protein, metal-rich granules, and heat-stable protein were determined by atomic absorption spectrometry. Exposure of C. pyrenoidosa to copper and cadmium in combination inhibited growth more strongly than copper and cadmium individually. Highest accumulation was observed in metal-rich granules and heat-stable proteins. Administration of both metals in combination affected their subcellular distribution: copper was mainly distributed into the metal-rich granules (70%–80%) and heat-stable proteins (9%–24%), cadmium in the metal-rich granules (88%–98%).     

In vivo accumulation of radioactive polonium by the giant kelp Macrocystis pyrifera

The in vivo accumulation rate of ²¹⁰Po by Macrocystis pyrifera (L) Ag. has been determined by the tagging of specific parts of young, growing lamina in La Jolla kelp beds and their subsequent collection for ²¹⁰Po analysis. The rates of accumulation for observed and growth-corrected data were, respectively, 0.78x10^-9 pCi/cm² sec (1.73x10^-9 dpm/cm² sec) and 1.17x10^-9 pCi/cm² sec (2.60x10^-9 dpm/cm² sec). Invert, but biologically foulable, glass-slide surfaces exposed to the kelp bed environment yielded ²¹⁰Po accumulation rates of 0.64x10^-9 pCi/cm² sec (1.42x10^-9 dpm/cm² sec), slightly less than those of M. pyrifera. Distinction is made between observed net accumulation rate and gross rates or total flux. Several factors contributing to the final net accumulation rate are discussed.     

The Influence of Varying Algal Biomass On Contaminant Exposure in Benthic-Planktonic Mesocosms: Copper (Ii)

Abstract

A series of mesocosms was exposed to a suite of light treatments and nutrient enrichment in order to generate algal communities of varying biomass. the influence of this biomass on the speciation of copper (II) was studied. Distribution coefficients ( _Kd,Lkg?1) were relatively high (log _Kd = 5 to 7), indicative of robust trace metal sequestration, and were likely controlled by the particulate organic carbon content (foc). Differences in _Kd over time and among treatments were significant, as was the relationship between _Kd and foc. Fluorescence quenching was used to determine binding capacities ( _Lt , M) and their associated binding constants ( _Kcond ,M^?1) in order to model the solid phase copper speciation. the _Kcond ranged between 2.1 and 5.2 × 10¹²M^?1, indicating a very strong copper-ligand complex, and was higher in mesocosms that received more light. the light _Lt increased over time, dramatically after the nutrient enrichment, but did not vary systematically among light treatments. _Lt ranged from 7.2 × 10? 7 to 4.9 × 10? 5 M. the large magnitudes of _Kd , _Kcond and _Lt ensured that greater than 97% of total copper in the mesocosms was complexed by organic matter. the total copper concentration ([Cu]_T, M) needed to reach a target dissolved copper concentration of 10^?12.5 M (pCu = 12.5) was determined for each mesocosm over time. [Cu]_T was between 8.02 × 10^?5 and 3.41 × 10^?2 M, and increased over time. the [Cu]_T normalized to the target pCu (Effective Dose Ratio, EDR) increased directly with increases in algal biomass, indicating a direct link between system productivity and copper exposure. Approximately 45% of the variance in EDR was explained by variance in total biomass, while the residual variance in EDR was due likely to differences in the strengths of particle associations and magnitude of binding capacities.     

Behavioural responses of the marine bivalve Macomona liliana exposed to copper- and chlordane-dosed sediments

Sublethal behavioural responses including avoidance and burial rate were compared with 10 d acute morbidity and mortality in bioassays using juvenile Macomona liliana 1 to 3 mm long. The bivalves showed significant movement away from all copper-dosed sediments, with maximal movement after 96 h at 25 mg Cu kg (dry wt)^-1. Increasing copper concentrations slowed the rate of burial, and above 15 mg Cu kg (dry wt)^-1, most shellfish failed to bury after 90 min. After 10 d exposure, morbidity (defined as inability to rebury) occurred at 15 mg Cu kg (dry wt)^-1 and mortality at 30 mg Cu kg (dry wt)^-1. In avoidance trials, chlordane had a significant effect on overall movement from 20 g kg (dry wt)^-1, but increasing concentrations reduced movement away from dosed sediment. Chlordance appeared to have no effect on burial rate. Survival was affected by 10 d exposure to 400 g chlordane kg (dry wt)^-1, with increased morbidity and death. The relative sensitivities of the responses shown by M. liliana to copper were avoidance > burial/morbidity > mortality, with a 6-fold difference in the response threshold between avoidance and mortality. With chlordance, an avoidance response was detected at a concentration 20-fold lower than that causing morbidity. Behavioural responses of M. liliana have the potential to provide a sensitive bioassay.     

Effects of calcium,strontium, and magnesium on the coccolithophorid Cricosphaera (Hymenomonas) carterae. I. Calcification

The present investigation concerns the effects of calcium, strontium, and magnesium on calcification and mineralogy of the calcified bodies (coccoliths) of the coccolithophorid Cricosphaera (Hymenomonas) carterae. The capacity of cells to calcify in various concentrations of these ions was examined following preliminary decalcification in CO₂. At a concentration of 10^-2 M Ca, 75% of the cells formed coccoliths within 24 h and almost all cells were recalcified after 2 days. At 10^-3 or 10^-4 M Ca no recalcification occurred. However, with the addition of Sr to the Ca-deficient media, calcification took place as shown by observations of coccoliths and by analysis of Ca. The percentage of calcified cells increased with increasing concentrations of Sr. Strontium added to a Ca-deficient media was much more effective than an equivalent concentration of Ca. No Sr was deposited in the coccoliths. X-ray analysis demonstrated that calcite was deposited by cells in all concentrations of Ca and Sr at which calcification took place. At concentrations of Mg in the media from O. O to 4.2×10^-2 M, the cells retained their ability to calcify, although calcification was markedly reduced in the absence of Mg. In low Mg concentrations (1.3×10^-4 and 4.2×10^-6 M), the coccoliths were 60% calcite and 40% aragonite, and in teh absence of Mg, only calcite was formed.     

Effect of calcium and magnesium on copper,cadmium, and some essential elements of Indian major carps Labeo rohita (Hamilton) and Catla catla (Hamilton) treated with copper and cadmium

The effect of calcium and magnesium either singly or in combination on accumulation of cadmium and copper in Labeo rohita (rohu) and Catla catla (catla) was investigated in this study under laboratory conditions. The investigation showed that copper accumulation in rohu exposed to 0.25 mg L^?1 of copper for 14 days reduced from 31.0 ± 0.4 mg kg^?1 at no calcium/magnesium treatment to 3.5 ± 0.2 and 2.2 ± 0.1 mg kg^?1, respectively, at 75 mg L^?1 calcium or magnesium treatment. The copper level in catla exposed to 0.20 mg L^?1 of copper for 14 days reduced from 5.7 ± 0.1 mg kg^?1 at no calcium/magnesium treatment to 3.4 ± 0.2 and 3.3 ± 0.1 mg kg^?1, respectively, at 300 mg L^?1 calcium or magnesium treatment. The cadmium accumulation in rohu exposed to 0.20 mg L^?1 of cadmium for 14 days reduced from 1.7 ± 0.1 mg kg^?1 at no magnesium treatment to 1.4 ± 0.1 mg kg^?1 at 120 mg L^?1 magnesium treatment. The cadmium accumulation in catla exposed to 0.20 mg L^?1 of cadmium for 14 days reduced from 0.8 ± 0.2 mg kg^?1 at no magnesium treatment to 0.6 ± 0.2 mg kg^?1 at 80 mg L^?1 magnesium treatment. Copper and cadmium treatments also reduced some essential microelements of rohu and catla. Both the fishes restored these elements at different levels of calcium and magnesium.     

Bioaccumulation and toxic effects of copper in common onion Allium cepa L.

The aim of this study was to investigate the potential utility of Allium cepa L. as a bioindicator organism for measuring copper bioaccumulation and toxicity in laboratory conditions. Onions were exposed to increasing concentrations of the metal (0, 0.1, 0.5, 1, 5 and 10 μg mL^?1) for 7 days. Root and leaf development were chosen as biological endpoints, while bioaccumulation was evaluated in roots, bulbs and leaves. Copper caused inhibition of root elongation with increasing effects at the higher doses, growth being reduced by almost 60% at 0.1 μg mL^?1 and up to 95% at 10 μg mL^?1. The elongation of leaves was significantly lower only in specimens exposed at 0.5 μg mL^?1, but a total absence of newly formed tissues was observed at 10 μg mL^?1. A marked bioaccumulation of copper was measured in roots, with values increasing up to almost four orders of magnitude compared to controls; only slight or even no significant differences were observed for copper levels in leaves and bulbs of treated A. cepa. Multiple linear correlations revealed a significant inverse relationship between copper concentrations and tissue length in both the roots and leaves, evidencing a sensitive responsiveness of this biological model. The overall results suggest the suitability of A. cepa as a robust species for easy and simple ecotoxicological bioassays to test the toxic effects and bioavailability of environmental pollutants, especially trace metals.     

Availability of copper from phytoplankton and water for the bivalveMacoma balthica. II. Uptake and elimination from64Cu-labelled diatoms and water

The amount of copper taken up via algae and water byMacoma balthica from the Oosterschelde sea arm, S.W. Netherlands, was established using the radioisotope⁶⁴Cu. As far as we know, this isotope has never been used before in marine flod chain studies. As a model food source the marine diatomPhaeodactylum tricornutum was allowed to accumulate⁶⁴Cu for 1 d. These labelled algae were fed to the clams in the presence of the complexing agent EDTA (0.27mM). EDTA was added to prevent uptake of dissolved⁶⁴Cu that could be leaking from the labelled diatoms. In control experiments, unlabelled diatoms were fed toM. balthica in the presence of dissolved⁶⁴Cu (with and without EDTA) in order to assure a similar filtration activity. In repeated experiments with varying particulate/dissolved copper ratios, uptake through food always turned out to be at least as efficient as uptake from the water. It was concluded that Cu, associated with food, is well available for uptake byM. balthica.     

Bioaccumulation and elimination of copper in the rock oyster Crassostrea cucullata

The kinetics of copper bioaccumulation in the rock oyster Crassostrea cucullata Born showed that the initial rate of uptake was directly related to metal concentration in the medium. As the accumulation in the tissues increased, the oysters remained closed and the uptake rate fell. At the end of 7 weeks, the average copper concentrations in the tissue were 60.42 g g^-1 and 63.97 g g^-1 wet weight in the 0.01 and 0.05 ppm experimental groups, respectively. The net rate of uptake ranged from 1.76 to 1.97 g g^-1 week^-1 and the rate of copper loss, measured after transferring the oysters into natural sea water, was dependent on the original cooper concentration in the soft parts. The concentration of copper in the tissues declined by 37.38 and 36.56% in the 0.01 and 0.05 ppm experimental groups, respectively. Even after a 7 week period of depuration (self-purification) there was some residual copper left in the tissue. This indicates that accumulation occurs in the tissue more rapidly than cleansing can eliminate it.     

Degradation of bacteria by Mytilus edulis

The uptake of several species of bacteria by the common mussel Mytilus edulis (L.) and the subsequent fate of some polymers of the bacteria have been investigated in a study carried out during 1981. Bacteria (Escherichia coli, Micrococcus luteus, M. roseus, Bacillus cereus, Staphylococcus aureus and a marine pseudomonad, 1-1-1) were radiolabelled by growth in medium containing ³H-thymidine and the uptake of bacteria by Mytilus edulis was monitored. Labelled and unlabelled bacteria, at initial concentrations of 0.5 to 1x10⁷ bacteria ml^-1, were cleared at similar, exponential rates with no significant difference in the rates for different bacteria: 90% of bacteria were cleared in a mean time of 1.93±0.12 h (SEM, n=63). Those bacteria with cell walls which were sensitive to M. edulis lysozyme were rapidly degraded by the mussel and ³H-labelled DNA was released in a form not precipitable by 10% trichloroacetic acid. Lysozyme-resistant bacteria (Micrococcus roseus and S. aureus) were cleared from suspension by Mytilus edulis but most were rejected intact. By measuring the rate of release of ³H-thymidine-labelled material from the mussel the rate of degradation of lysozyme-sensitive bacteria by M. edulis was found. For different bacteria the degradation rate varied from approx 2x10⁸ to 27x10⁸ bacteria h^-1 with an overall mean of 10x10⁸ bacteria h^-1. A thymidine- and diaminopimelicacid-requiring auxotroph of E. coli was radiolabelled with ³H-thymidine, ³H-diaminopimelic acid or ¹⁴C-glucose and fed to M. edulis. Bacteria were cleared and degraded by the mussel; ³H-diaminopimelic acid-labelled or ¹⁴C-glucose-labelled polymers were retained, whereas ³H-thymidine-labelled polymers were released into the surrounding water. Extracts of the digestive gland of M. edulis degraded lysozyme-sensitive bacteria to release ³H-thymidine-labelled material, but did not release ³H-thymidine-labelled material from lysozyme-resistant bacteria. It is concluded that M. edulis can select lysozymesensitive bacteria for subsequent processing and discriminate between bacterial polymers to reject DNA. Also, bacteria could provide a substantial fraction of the carbon requirement of the mussel.     

The biological degradation of arginine and glutamic acid in seawater in relation to the growth of phytoplankton

The responses of natural summer coastal plankton communities to low-level additions (10^-5 to 10^-7 M) of arginine and glutamic acid has been followed by in vivo measurement of chlorophyll fluorescence. This technique is capable of detecting a response to a 10^-6 M enrichment under most conditions. The time sequence of the response varied with the amino acid used and with the enantiomeric form of the amino acid. Ammonia and carbon dioxide were liberated before the increase in chlorophyll fluorescence occurred. Liberation of ammonia in a dark bottle from L-arginine was from 75 to 85% of the theoretical yield. Microautoradiography using ¹⁴C L-arginine or L-glutamic acid at 10^-7 M showed heavy labeling associated with fecal pellets and detrital aggregates. Phytoplankton cells were not appreciably labeled. The evidence suggests that bacteria are important in the cycling of these compunds.     

Synergistic physiological effects of low copper and various oxygen concentrations on Macoma balthica

The negative effects of copper on Macoma balthica are significantly increased when this bivalve is simultaneously exposed to low oxygen concentrations. This might be explained by the fact that low oxygen concentrations (oxygen deficiency) are combined with a slightly lower pH and thus with a different copper speciation, resulting in a higher bio-availability of copper and, consequently, in a higher copper uptake by M. balthica. Symptomatic for the increased negative effects of copper on M. balthica at low oxygen concentrations is an increased oxygen demand, which in turn reflects an increased consumption of energy. Consequently, at low oxygen levels, low copper concentrations (8–15 μg dm^-3) greatly affect the glycogen content of the tissues, and finally (at 15 μg Cu dm^-3) result in a heavy loss of dry weight and a drastic reduction of the adenylate energy charge (AEC). Generally, it might be concluded that dry weight, glycogen content and AEC are more affected by a combination of oxygen deficiency and a low copper concentration (2.5 cm³ O₂ dm^-3, 15 μg Cu dm^-3) than by higher copper concentrations (up to 86 μg dm^-3) at high oxygen levels (5.0 cm³ dm^-3).