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.     

Bioavailability of dissolved copper to the American oyster Crassostrea virginica. I. Importance of chemical speciation

The effect of complexation on the accumulation of dissolved copper by the American oyster Crassostrea virginica was determined in chemically defined exposure media. The speciation of copper was varied by varying the concentrations of total copper and model chelator, nitrilotriacetic acid (NTA). Accumulation of copper in 14-d experiments was related to the cupric ion activity and not the concentration of chelated copper. Rapid accumulation of copper occurred at cupric ion activities above 10^-10 M and there was no measurable accumulation at 10^-11 M.Southeast Fisheries Center Contribution No. 81-45B. Reprint request should be addressed to this author     

Effects of Olisthodiscus luteus on the growth and abundance of Tintinnids

The effects of the red tide flagellate Olisthodiscus luteus Carter on the growth of two tintinnid species, Tintinnopsis lubulosoides Meunier and Favella sp. (Clap. & Lach.) Jorg., were measured in batch culture. T. tubulosoides and Favella sp. grew at rates equivalent to 1.2 (10°C) and 2.0 (20°C) population doublings per day, respectively, when offered nutritionally adequate phytoplankton species. The growth rates of both tintinnid species were reduced in the presence of 10²–10³ O. luteus cells · ml^-1 in multialgal treatments. Growth rate inhibition was proportionately greater at higher O. luteus densities. Lethal effects were observed for both tintinnid species at O. luteus concentrations of 5x10³ cells · ml^-1 in multi-algal treatments. T. tubulosoides mortality occurred at all O. luteus concentrations in unialgal culture. O. luteus-conditioned medium did not substantially inhibit tintinnid growth when combined with acceptable food species, suggesting that toxicity is induced by ingestion or direct contact with O. luteus cells, or by exposure to a short-lived exudate. In agreement with these results, an inverse relationship between O. luteus concentration and tintinnid abundance was observed in Narragansett Bay, Rhode Island, over a two year period. The small lorica diameter of the species apparently inhibited by these O. luteus blooms suggests a detrimental effect independent of cell ingestion. In addition to the absolute concentration of O. luteus cells, the availability of nutritionally adequate algal food may be an important factor determining the impact of O. luteus blooms on tintinnid populations.Contribution no. 5048 from the Woods Hole Oceanographic Institution     

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.     

Growth and grazing rates of the tintinnid ciliate<Emphasis Type="Italic"> Favella taraikaensis</Emphasis> on the toxic dinoflagellate<Emphasis Type="Italic"> Alexandrium tamarense</Emphasis>

Growth and feeding activities of the tintinnid ciliate Favella taraikaensis fed the toxic dinoflagellate Alexandrium tamarense were examined in laboratory experiments. Both growth and ingestion rates of F. taraikaensis as a function of the A. tamarense concentration were fitted to a rectangular hyperbolic equation. The maximum growth and ingestion rates of F. taraikaensis were 1.0 day^–1 and 2.8 cells ind. h^–1 (carbon specific ingestion rates: 3.5 day^–1), respectively, which are both included in the range of previous data reported for Favella spp. feeding on other algae. The gross growth efficiency (GGE) of F. taraikaensis ranged from 0.26 to 0.49 (mean value 0.40) at the concentration of 10–800 cells ml^–1, which is within the range of previous data on Favella spp. Also, the growth and ingestion rates and GGE of F. taraikaensis on A. tamarense were not significantly different from the values on another non-toxic dinoflagellate (Heterocapsa triquetra) at two different prey concentrations. This indicates that the toxicity of A. tamarense probably did not influence the feeding and growth activities of F. taraikaensis at concentrations of less than ca. 800 cells ml^–1. To evaluate the grazing by F. taraikaensis on A. tamarense blooms in the field, the population dynamics of A. tamarense were simulated based on the growth and ingestion parameters of F. taraikaensis. As a result, the grazing impact by F. taraikaensis was considered to potentially regulate the development of A. tamarense blooms. If the toxicity of A. tamarense does not influence the growth and feeding activities of F. taraikaensis, the occurrence of such grazer plankton are considered to be important for predicting the course of A. tamarense bloom dynamics under natural conditions.Communicated by T. Ikeda, Hakodate     

Glycolate metabolism by Pseudomonas sp., strain S227, isolated from a coastal marine sediment

Glycolate excreted by phytoplankton is a potentially important nutrient for bacteria in coastal and estuarine environments. The metabolism of glycolate by Pseudomonas sp., strain S227, originally isolated from the New York Bight Apex, has been studied. The specific growth rate for this strain on glycolate is 0.156 doublings h^-1. The apparent V_max and K_m for glycolate uptake are 83.6 nmol min^-1 mg cell protein^-1 and 7.4x10^-8 M, respectively. The preferential respiration of the carboxyl carbon (C-1) and the incroporation of the hydroxymethyl carbon (C-2) suggest that the glycerate pathway is used for growth on glycolate. Alternatively, another pathway can be utilized which results in the complete catabolism of glycolate. Glycolate and lactate metabolism are also closely linked either by a common metabolic pathway or a common transport system other than the monocarboxylate transport system. The magnesium ion concentration is also important in glycolate metabolism. The characteristics of glycolate metabolism observed in Pseudomonas sp., strain S227, are advantageous in coastal and estuarine environments where glycolate production is intermittent, and the concentrations are low.     

Contribution to the ecotoxicological study of cadmium,copper and zinc in the mussel Mytilus edulis

A regulation of internal levels of some essential metals has been observed in various animals, whereas the bioaccumulation of several non-essential metals parallels their overloads in water. In the mussel Mytilus edulis L., we have attempted to determine if such a phenomenon exists by comparing the patterns of accumulation of copper and zinc vs cadmium. With this aim, mussels collected in the Bay of Bourgneuf (France) in November 1983 were exposed to these metals for 16 d. At external levels of zinc as high as 100 gl^-1, mussels were able to maintain a normal concentration in all groups of organs for 4 d. The ability of mussels to limit the bioaccumulation of copper and zinc varied from organ to organ, and decreased with higher levels of contamination and longer periods of exposure. In contrast, at the lowest experimental concentration and the lowest period of exposure, a significant increase of cadmium in mussel tissues was generally observed. Even at the highest sub-lethal doses, the levels of copper and zinc in mussel tissues were not much higher than the natural levels (contaminated:background ratios= 2.3 to 6.1), whereas the bioaccumulation of cadmium was less well restricted (contaminated:background ratios=136 to 192). The use of mussels as a bioindicator of pollution seems doubtful for essential metals, particularly as regards short-term pollution, since the levels of these trace elements in the organisms are largely independent of their concentration in the ambient seawater.     

Toxicity of zinc,cadmium and copper to the shrimp Callianassa australiensis. I. Effects of individual metals

The acute toxicity of zinc, cadmium and copper to Callianassa australiensis (Dana) was evaluated in static tests. Each test lasted up to 14 d and LC₅₀ values were calculated for 4, 7, 10 and 14 d intervals. The toxicity of each metal increased with exposure time; thus the 4 d LC₅₀ values of 10.20, 6.33 and 1.03 mg l^-1 were considerably higher than the 14 d LC₅₀ values of 1.15, 0.49 and 0.19 mg l^-1 for zinc, cadmium and copper respectively. Toxicity curves reveal that none of the values were asymptotic, indicating that median lethal threshold concentrations were not reached for any of the metals. This suggests that 14 d is an insufficient time in which to complete meaningful, acute lethality tests for marine shrimps. Longer tests are necessary if lethal threshold concentrations are to be used with application factors to derive safe concentrations for the protection of C. australiensis.     

Toxic effects of heavy metals upon cultures of Uronema marinum (Ciliophora: Uronematidae)

The lethal levels of mercuric, lead and zinc chlorides were determined for the marine ciliated protozoan Uronema marinum Dujardin; the 24 h LC₅₀ values were 0.006, 60 and 400 mg l^-1, respectively. A factorial experiment was used to study the inhibition of cell division in U. marinum cultures by mixtures of mercury, lead and zinc. The three metals were found to interact in their effects. A regression model was fitted to the experimental results and the response surface was plotted. The addition of mercury, lead or zinc at concentrations which inhibited cell division did not appear to affect the duration of the lag phase or the yield of cells. A culture of U. marinum was maintained for 18 weeks in a medium containing mercury, lead and zinc in an attempt to acclimatise the ciliates to heavy metals. A subsequent factorial experiment which examined the cell division rate indicated that a prolonged exposure did not induce in the ciliates a measurable tolerance to the metals.     

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. 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.     

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.     

The nature of zinc and copper complexes in the oyster Ostrea edulis

The zinc and copper associated with the soft tissues of the oyster Ostrea edulis Linnaeus have been separated into a soluble component and a tissue-residue, cell-debris bound component. In the case of zinc, the tissue-bound component was found to contain at least two species of complex; a firmly-bound species, exchangeable with ⁶⁵Zn²⁺ and a less-firmly, reversibly-bound species, exchangeable with ⁶⁵Zn²⁺. The soluble component, which constitutes some 40% of the total zinc and copper, was fractionated on Sephadex G-25 and the zinc and copper shown to be weakly-complexed to the small molecular weight compounds, taurine, lysine, ATP and possibly homarine (N-methyl--picolinic acid) and to be fully exchangeable with ⁶⁵Zn²⁺. These soluble complexes can act as a freely available mobile reserve of metal to ensure a constant saturation of metal-dependent enzyme systems operating under adverse environments. Sephadex G-25 acts as a weak ion-exchange resin, which can cause a translocation of zinc and copper from its soluble weak complexes and result in the spurious association of the metals with other compounds.     

Biodegradation kinetics of cymoxanil in aquatic system

A potential method to detoxify pesticides in aquatic system is using bioremediation. In this study, four microorganisms (Pseudomonas sp (EB11), Streptomyces sp. (EB12), Aspergillus niger (EB13) and Trichoderma viride (EB14) were isolated from cucumber leaves previously treated with cymoxanil using enrichment technique. These strains were evaluated for their potential to detoxify cymoxanil in aquatic system at the concentration level of 5×10^?4M. The effect of pH and temperature on the growth ability of the tested strains was also investigated by measuring the intracellular protein and mycelia dry weight for bacterial and fungal strains, respectively. Moreover, the remaining toxicity of cymoxanil after 28 days of incubation with tested strains was evaluated to confirm the complete removal of any toxic materials (cymoxanil and its metabolites). The results showed that the optimum pH for the growth of cymoxanil degrading strains (bacteria and fungi) was 7. A temperature of 30°C appears to be the optimum for the growth of either fungal or bacterial strains. Pseudomonas sp. (EB11) was the most effective strain in cymoxanil degradation followed Streptomyces sp (EB12), Trichoderma viride (EB14) and Aspergillus niger (EB13), with half-lives of 4.33, 9.5, 17.3 and 24.7 days, respectively. The degradation of cymoxanil by bacterial strains was much faster than fungal one. There is no remaining toxicity of cymoxanil detected in aqueous media previously treated with Pseudomonas sp. (EB11) for 28 days. The results suggest that bioremediation by Pseudomonas sp. (EB11) are promising for the detoxification of cymoxanil in aqueous media.     

Effects of algal and larval densities on development and survival of asteroid larvae

Effects of larval and algal culture density and diet composition on development and survival of temperate asteroid larvae were studied in the laboratory at Santa Cruz, California, USA, during summer and fall of 1990. Larvae of Asterina miniata were reared at two densities, 0.5 or 1.0 ml^-1, and fed one or two species of cultured phytoflagellates — Dunaliella tertiolecta alone or mixed with Rhodomonas sp. — at three concentrations of 5x10², 5x10³, and 5x10⁴ total cells ml^-1. Algal concentration strongly influenced larval development; however, larval density also had a marked effect. Development progressed further with increasing algal concentration. Larval growth and differentiation were sometimes uncoupled; i.e., growth measures were directly related to food level, while differentiation indicators were less so. At the lowest food level, growth was negative and differentiation was arrested at early precompetent stages; these larvae never formed juvenile rudiments or brachiolar attachment structures. Development times of larvae given more food ranged from 26 to 50 d and depended directly on food availability. Development time to metamorphosis at the highest food concentration was similar for siblings fed D. tertiolecta alone or mixed with Rhodomonas sp. In contrast, when food level was an order of magnitude lower, larvae fed the algal mixture metamorphosed significantly earlier than larvae fed the unialgal diet. This suggests interactive effects of food quantity and food quality. Survival was little affected by larval or food density, except at the lowest ration. Feeding experiments in well-controlled laboratory conditions are useful to predict and compare the physiological or developmental scope of response of larvae to defined environmental factors; however, results from such studies should not be extrapolated to predict rates and processes of larval development in nature.     

Ecological growth strategies in the seaweeds Gracilaria foliifera (Rhodophyceae) and Ulva sp. (Chlorophyceae): Soluble nitrogen and reserve carbohydrates

The seaweeds Gracilaria foliifera (Rhodophyceae) and Ulva sp. (Chlorophyceae) were grown in an outdoor continuous-flow system at both ambient incident light (I₀) and 0.13 I₀. During the winter, both species accumulated substantial soluble nitrogen reserves (up to 1020 g-at N·g dry wt^-1 in G. foliifera and 630 g-at N·g dry wt^-1 in Ulva sp.). The rate at which these N reserves were depleted was proportional to the growth rate. Seaweeds grown at 0.13 I₀ had lower growth rates and higher levels of soluble tissue N than plants grown at I₀. During the spring-summer growing season, peaks in tissue N followed nutrient peaks in the ambient seawater. Ulva sp. had higher nutrient uptake and growth rates than G. foliifera and showed greater fluctuations in soluble tissue N. This may characterize opportunistic seaweed species with high biomass turnover rates. At I₀, the levels of starch (up to 340 mg·g dry wt^-1 in G. foliifera and 170 mg·g dry wt^-1 in Ulva sp.) were highest during the spring and summer. During this period, fluctuations in starch content were inversely related to growth rate and soluble tissue N. Seaweeds grown at 0.13 I₀ did not accumulate starch. Neither species was found to overwinter with starch reserves.     

Influence of relative water motion on the growth,ammonium uptake and carbon and nitrogen composition of Ulva lactuca (Chlorophyta)

In a series of multifactorial laboratory experiments, Ulva lactuca discs were grown in an apparatus in which they were exposed simultaneously to 3 simulated current speeds (7.5, 15, 22.5 cm s^-1) and a still control, and either 3 ammonium concentrations (0–10, 35–45 and 115–145 M) under ample uniform light (ca 200 E m^-2 s^-1) or 3 light intensities (approximately 35, 90 and 270 E m^-2 s^-1) with uniform surplus, ammonium. Disc growth rates were determined in each experiment as well as tissue nitrogen and carbon composition and fluxes of NH₄, NO₃/NO₂ and PO₄ in media. In a supplementary series of field experiments, U. lactuca discs were simultaneously exposed to 2 different water motion regimes in adjacent chambers at several sites characterized by widely different ammonium concentrations. In field experiments, growth rates were calculated and analyzed as a function of water motion at the various sites. The application of simulated current consistently enhanced disc growth rates in the laboratory, except at the lowest light intensity. In most cases this enhancement was fully realized at the lowest applied simulated current (7.5 cm s^-1). Simulated current slightly enhanced ammonium uptake rates by U. lactuca discs, relative to rates in still water, except at the highest ammonium concentration. C:N ratios of discs generally declined with increases in simulated current, except at the highest ammonium concentration. This decline was primarily attributable to increases in per cent N and was, again, mainly realized at 7.5 cm s^-1. The results suggested that simulated current compensated for N limitation, except when light was sufficiently low to become the overriding limiting factor, but that the enhancement of growth by simulated current could not be explained in terms of N metabolism alone. Field experiments showed that the higher level of water motion consistently enhanced growth at sites with comparatively low ammonium concentrations, but not at sites with moderate or high ammonium concentrations.     

Microphagous ciliates in mesohaline Chesapeake Bay waters: Estimates of growth rates and consumption by copepods

Growth rates of microphagous ciliates (forms which feed primarily on picoplankton-sized prey) were estimated, along with rates of their consumption by copepods, in shipboard experiments conducted in the mesohaline portion of Chesapeake Bay, USA, under contrasting water column conditions in April, June, and August 1987. Estimates were based on temporal changes in cell densities in size-fractionated water samples incubated under in situ conditions. In April, at low temperatures (7 to 10°C) and with oxygen present throughout the water column, similar generation times of ca. 1 to 1.5 d were estimated for surface and deep water (24 m) ciliate populations. In June, water was anoxic below 12 m and a distinct anoxic microphage community grew at about twice the rate of the surface community with generation times of ca. 7 and 14 h, respectively. In August, bottom water was again anoxic, but the sameStrobilidium sp. dominated both surface and deep waters with low or no growth apparent in anoxic waters and a generation time of ca. 8 h in surface waters. Copepod (primarilyAcartia tonsa Dana nauplii) clearance rates for microphagous ciliates in surface waters were 0.11, 0.56, and 0.53 ml h^–1 copepod^–1 for April, June and August, respectively. Calculation of removal rates, based on average densities, indicated that from 34 to 200% of surface waters were cleared d^–1 of microphagous ciliates by copepods.     

Light absorption,photosynthesis and growth of Nannochloris atomus in nutrient-saturated cultures

Nannochloris atomus was maintained in exponential growth at photon flux densities (PFD) from 400 to 700 nm, ranging from 10 to 200 mol m^-2 s^-1. Growth was lightsaturated at PFDs in excess of 100 mol m^-2 s^-1, with a mean light-saturated growth rate at 23 °C of 1.5×10^-5s^-1 (1.2 d^-1). The light-limited growth rates extrapolated to a compensation PFD for growth that was not significantly different from zero, although no changes in cell numbers were observed in a single culture incubated at a PFD of 1.0 mol m^-2s^-1. Dark-respiration rates were independent of PFD, averaging 1.7×10^-6 mol O₂ mol^-1 C s^-1 (0.14 mol O₂ mol^-1 C d^-1). The maximum photon (quantum) efficiency of photosynthesis was also independent of PFD, with a mean value of 0.12 mol O₂ mol^-1 photon. The chlorophyll a-specific light absorption cross-section ranged from 3 to 6×10^-3 m² mg^-1 chl a and was lowest at low PFDs due to intracellular self-shading of pigments associated with high cell-chlorophyll a contents. The C:chl a ratio increased from 10 to 40 mg C mg^-1 chl a between PFDs of 14 and 200 mol m^-2 s^-1. These new observations for N. atomus are compared with our previous observations for the diatom Phaeodactylum tricornutum in terms of an energy budget for microalgal growth.