Transport of naphthalene in the oyster Ostrea edulis

In small oysters (Ostrea edulis), transport of naphthalene between tissues is primarily by diffusion and not via the circulatory system. In intact oysters, accumulation in the adductor muscle and body followed accumulation in the gills after a large lag-time. In isolated tissues with no shell to impede water flux over the body and adductor muscle, there was no lag-time. The molecular diffusivity (D) of naphthalene in oyster tissue, estimated by Fick's second law of diffusion is D=8x10^-8 cm² s^-1, a value similar to D determined for lateral diffusion of lipophilic compounds in lipid membrane systems.     

Coral calcification: Sources of error in radioisotope techniques

Isotopic exchange occurs between coral skeleton and ⁴⁵Ca⁺⁺ and H¹⁴CO ₃ ^- in seawater. Exchange of ¹⁴C onto skeletons is more rapid than exchange of ⁴⁵Ca⁺⁺. Exchange of ¹⁴C from skeletons to seawater takes place more slowly than exchange of ⁴⁵Ca⁺⁺ to seawater. When living coral is incubated in the dark with radioisotopes for 1 h, the tissues contain considerably more radioactivity than is associated with the skeleton. The tissue radioactivity reflects permeation of tissues and coelenteron by radioactive compounds from the incubation seawater. Addition of alkalis to cardioactive seawater results in a radioactive precipitate, part of which becomes associated with any coral skeleton present, and part of which forms on the wall of the containing vessel. Strong alkali removes biologically-deposited radioisotope from coral skeletons. Deposition, of ¹⁴C from H¹⁴CO ₃ ^- in skeletons of living coral incubated in the dark is greater than in dead coral. The reverse situation occurs with ⁴⁵Ca⁺⁺.     

Possible causes of temporal fluctuations in primary production of the microphytobenthos in the Danish Wadden Sea

The effects of prior exposure to unlabelled naphthalene on the processes of uptake, tissue distribution and elimination of [1-¹⁴C] naphthalene by mussels (Mytilus edulis) were examined. Mussels collected from Whitsand Bay (S. W. England) in October 1980 were either held unexposed or exposed to unlabelled naphthalene (0.5 μg 1^?1) for 4 wk, prior to receiving a 4 h pulse of [1-¹⁴C] naphthalene in the medium (7 μg 1^?1). After 4 h exposure to [1-¹⁴C] naphthalene, the major tissues (digestive gland, gills, kidneys, mantle and remaining tissue) showed exponential depuration curves which could be resolved into two statistically significant components. The kinetics of elimination and the biological half-times of [1-¹⁴C] naphthalene in the tissues were determined. Mussels pre-exposed to unlabelled naphthalene had a significantly higher rate of elimination of [1-¹⁴C] naphthalene from the gills and kidneys. These results indicate that the toxicant, naphthalene, is actively excreted from the body by the gills and kidneys and the process is enhanced by prior exposure to the toxicant.     

Preparation and evaluation of protein microcapsules for a marine suspension-feeder,the Pacific oyster Crassostrea gigas

Little is known of the dietary requirements of marine suspension-feeders, mainly due to difficulties in preparing and delivering microparticulate diets to animals living in an aquatic environment. The food particles must be stable in seawater and digestible by the consumer. A laboratory method for preparing cross-linked, protein-walled capsules is described in this study as a means of delivering dietary protein to marine suspension-feeders such as the oyster Crassostrea gigas. Leakage of ¹⁴C-labelled protein from capsules suspended in seawater for 24 h was reduced to less than 5% by the use of the trifunctional cross-linking agent 1,3,5-benzenetricarbonyl trichloride in wall formation. Invitro experiments indicated that capsules were susceptible to digestion by proteases and by style extracts of oysters. Oysters fed encapsulated ¹⁴C-labelled protein absorbed ¹⁴C with an efficiency of 40.2±5.0% (±1 SE). Biochemical fractionation of capsule-fed oysters indicated breakdown of ingested ¹⁴C-protein into material that was soluble in 7% trichloroacetic acid. Cross-linked, protein-walled capsules can be used successfully to deliver dietary protein to the oyster C. gigas.     

Effects of anaerobiosis on root metabolism of Zostera marina (eelgrass): implications for survival in reducing sediments

The temperate seagrass Zostera marina L. typically grows in highly reducing sediments. Photosynthesis-mediated O₂ supplied to below-ground tissues sustains aerobic respiration during photosynthetic periods. Roots, however, experience daily periods of anoxia and/or hypoxia at night and under conditions that reduce photosynthesis. Rhizosphere cores of Z. marina were collected in August 1984 from Great Harbor, Massachusetts, USA. We examined short-term anaerobic metabolism of [U-¹⁴C]sucrose in excised roots and roots of intact plants. Under anaerobic conditions roots showed appreciable labeling of CO₂, ethanol and lactate, and slight labeling of alanine and other metabolites. Over 95% of the ¹⁴C-ethanol was recovered in the root exudate. Release of other metabolites from the roots was minimal. Ethanol was also released from hypoxic/anoxic roots of intact plants and none of this ethanol was transported to the shoot under any experimental conditions. Loss of ethanol from roots prevented tissue levels of this phytotoxin from increasing during anaerobiosis despite increased synthesis of ethanol. Anaerobic metabolism of [U-¹⁴C]glutamate in excised roots led to appreciable labelling of -aminobutyrate, which was known to accumulate in eelgrass roots. Roots recovered to fully aerobic metabolism within 4 h after re-establishment of aerobic conditions. The contributions of these root metabolic responses to the ability of Z. marina to grow in reducing marine sediments are related to light-regulated interactions of shoots and roots that likely dictate depth penetration, distribution and ecological success of eelgrass.     

Uptake,metabolism and discharge of polycyclic aromatic hydrocarbons by marine fish

The uptake, metabolism and discharge of two polycyclic aromatic hydrocarbons, ¹⁴C-naphthalene and ³H-3,4-benzopyrene, were studied in 3 species of marine fish (mudsucker or sand goby, Gillichthys mirabilis; sculpin, Oligocottus maculosus; sand dab, Citharichthys stigmaeus). The path of hydrocarbons through the fish included entrance through the gills, metabolism by the liver, transfer of hydrocarbons and their metabolites to the bile, and, finally, excretion. The gall bladder was a major storage site of labeled hydrocarbons and their metabolites. The major product of ²H-3,4-benzypyrene metabolism was tentatively identified as 7,8-dihydro-7,8-dihydroxybenzopyrene. The ¹⁴C-naphthalene was metabolized to 1,2-dihydro-1,2-dihydroxynaphthalene after 24 h exposure. The urine appeared to the major avenne for discharge of labeled hydrocarbon from the body. Our laboratory results indicated that certain polycyelic aromatic hydrocarbons were rapidly taken up from seawater by the above fish, but detoxification mechanisms existed for efficient removal of these compounds from their body tissues.     

Crassostrea virginica as an indicator of lead pollution

After treatment with 1.0 and 3.3 g Pb kg^-1 for 20 weeks in flowing seawater at ambient salinity and temperature, Crassostrea virginica accumulated as much as 6.57 and 11.42 g g^-1 dry weight, respectively, and no mortalities were recorded. Lead uptake was curvilinear; however, the general trend was an increase in lead concentration over time. Dry weight of the oyster had no significant relation with tissue lead concentration (g g^-1); however, a significant positive relationship existed between weight and lead content (g). A decrease in lead concentration in the tissues occurred from mid-August to mid-September despite continuous addition of lead to the seawater. Addition of lead to the seawater was terminated after 20 weeks (October), and lead loss was studied for the following 12 weeks. Lead loss was studied well after spawning in an attempt to eliminate any influence this might have on lead concentration in the tissues. After 4 weeks, approximately 54% of the accumulated lead was lost under conditions of a natural temperature decline. The biological half-life of lead in C. virginica tissues was calculated to be 5.5 weeks. In the ensuing 8 weeks, little or no lead loss was observed. Larvae from each treatment developed to the straight-hinged stage within 48 h with no apparent abnormalities. Lead appeared to have no adverse effect on larvae when parents had been treated with 1.0 and 3.3 g kg^-1 seawater for 10 weeks.     

Calcification in the staghorn coral Acropora acuminata: Variations in apparent skeletal incorporation of radioisotopes due to different methods of processing

Pieces of branch from the staghorn coral Acropora acuminata were incubated with ⁴⁵CaCl₂ and NaH¹⁴CO₃ under identical conditions in the light or in the dark. Specimens were then processed in different ways. All specimens were placed in N KOH to digest tissues. Some were placed in KOH immediately after incubation; others were placed in KOH after 2 h washing, or after 2 h extraction with methanol-chloroformwater. Specimens were washed in running fresh water or running seawater; some were killed in liquid N₂ before washing. Radioactivity associated with skeleton and tissues was determined. The method of processing profoundly affected the results. In dark incubations, there was up to a four-fold difference in apparent skeletal incorporation of ⁴⁵Ca⁺⁺ between average values obtained for the different treatments. For ¹⁴C incorporation, there was a difference of up to 2.5 times. In light incubations, skeletal incorporation of both radioisotopes showed a two-fold difference between high and low average values obtained for the different treatments.     

Hydrocarbon metabolism in the intertidal fiddler crab Uca pugnax

The fiddler crab Uca pugnax was examined for its ability to metabolize foreign hydrocarbons. The microsomal mixed function oxidase system was identified in U. pugnax tissues using Aldrin epoxidation rates as the assay. Rates were slow: 96 pM Dieldrin per mg microsomal protein per hour in the hepatopancreas, 438 pM mg^-1 h^-1 in the gill, and 228 pM mg^-1 h^-1 in claw-muscle microsomes. Using standard methods, no difference in rates could be detected between crabs living in clean areas and those living in environments highly contaminated with foreign hydrocarbons. In vivo rates of naphthalene oxidation were measured and used to calculate a clearance time for U. pugnax body tissues based on the aromatic hydrocarbon content of crabs collected from an oil-polluted salt marsh. Calculated clearance time was beyond the life span of the crab. It is concluded that this minimal ability of U. pugnax to metabolize foreign hydrocarbons partially accounts for its sensitivity to oil pollution in the environment.     

Latitudinal patterns of shell thickness and metabolism in the eastern oyster Crassostrea virginica along the east coast of North America

The goal of this study was to quantify growth and metabolic responses of oysters to increased temperatures like those that will occur due to global warming. Impact of temperature on eastern oyster (Crassostrea virginica) shell growth and metabolism was investigated by sampling 24 sites along the eastern North American seaboard ranging from New Brunswick, Canada, to Florida, USA, in March and August 2013. There was a positive correlation between oyster shell thickness and site temperature. At southern sites, shells were up to 65 % thicker than at the northernmost site, likely due to higher precipitation of CaCO₃ in warmer water. This was supported by laboratory experiments showing that thicker shells were produced in response to temperatures 2, 4, and 6 °C above ambient seawater temperatures (8–14 °C) in Connecticut, USA. Field experiments with oyster respiration were conducted during winter and summer at 13 sites to compare responses to thermal stress with latitude. Respiration rates were much higher during summer than winter, but the combination of summer and winter data fell along the same exponential curve with respect to temperature. At all sites, temperature-specific metabolic rates at elevated temperatures were lower than predicted, indicating significant seasonal acclimatization by C. virginica.     

Exudation of biologically-active metabolites in the sponge Aplysina fistularis

Specimens of the demosponge Aplysina fistularis forma fulva (Pallas) from the intertidal zone at La Jolla, California, USA, contain two antimicrobial metabolites, aerothionin (1) and homoaerothionin (2). Both metabolites are exuded into seawater. The rates of exudation into artificial seawater were measured under near in situ conditions (7.7x10^-4 g aerothionin min^-1 g^-1 dry weight of sponge) and in an aquarium (8.9x10^-3 g min^-1 g^-1) for undamaged sponges. Sponges responded to a simulated injury by exuding 10 to 100 times as much of these metabolites. The exudation of aerothionin and homoaerothionin may inhibit surface overgrowth by fouling organisms and tissue damage by mobile animals.Please address requests for reprints to Dr. J. E. Thompson     

Zooxanthellae providing assimilatory power for the incorporation of exogenous acetate in Heteroxenia fuscescens (Cnidaria: Alcyonaria)

The soft coral Heteroxenia fuscescens (Ehrb.) and its isolated zooxanthellae (endosymbiotic dinoflagellates) were investigated with particular regard to uptake and utilization of exogenously supplied ¹⁴C-acetate in the light and in the dark. The incorporation of ¹⁴C from ¹⁴C-acetate into the host tissue and into the zooxanthellae was consistently much higher in the light than in the dark. The incorporated ¹⁴C-acetate was rapidly metabolized by the host and algae and was recovered from different assimilate fractions. The major proportion of radiocarbon from metabolized ¹⁴C-acetate was located in host tissue. The CHCl₃-soluble fraction composed of diverse lipids showed the strongest ¹⁴C-labelling. Zooxanthellae isolated prior to incubation accounted for about 80% of the acetate incorporation recorded for zooxanthellae in situ (in vivo). It is concluded from a comparison of acetate incorporation and conversion under light and dark conditions that most of the lipid reserve of the host tissue originates from fatty acids, which are synthesized within the algal symbionts and are then translocated to the heterotrophic partner via extrusion. The acetate units needed for lipid synthesis are obtained by absorption of free acetate from dissolved organic matter (DOM) in the seawater as well as by photosynthetic assimilation of inorganic carbon. Thus, in H. fuscescens, lipogenesis is operated as a light-driven process to which the zooxanthellae considerably contribute assimilatory power by performing fatty acid synthesis and translocation of lipid compounds to their intracellular environment (host cell). A metabolic scheme is proposed to account for the different pathways of carbon conversion observed in H. fuscescens. The incubations took place in August 1980 and the analytical part from October 1980 to January 1984.     

Carbon- and oxygen-isotope composition of the cuttlebone of Sepia officinalis: a tool for predicting ecological information?

Analysis of the isotope composition of calcareous structures of marine organisms has proved useful in providing biological data. The present study constitutes the first detailed work undertaken on the isotope composition of coleoid cephalopods. We analysed the carbon- and oxygen-isotope composition [δ¹³C (CO²⁻ ₃) and δ¹⁸O (CO²⁻ ₃), respectively] of the cuttlebone aragonite of wild and cultivated specimens of Sepia officinalis Linnaeus, 1758. δ¹³C (CO²⁻ ₃) ranged from −2.94 to 1.00‰, δ¹⁸O (CO²⁻ ₃) from −0.18 to 2.08‰. The carbon-isotope composition is not in equilibrium with the carbon species of the ambient seawater, and does not reflect the deposition of CaCO₃ in seawater. The potential influence of environmental factors and biological processes on the carbon-isotope composition of the cuttlebone is discussed. In contrast to δ¹³C, the oxygen-isotope composition of cuttlebone aragonite appears to be in isotopic equilibrium with the ambient seawater. Seasonal changes in isotopic temperature revealed by our analyses agreed with changes in the temperature of the ambient seawater. CaCO₃ was deposited all year round. A maximum life span of 2 yr, a year-round spawning season, and variable growth rates among and within individuals have been inferred from the isotopic temperatures. Received: 14 April 1998 / Accepted: 26 November 1998     

Biodegradation of aromatic hydrocarbon in marine sediments of three North Sea oil fields

On a survey around three North Sea oil fields in 1980 the biodegradation of the hydrocarbons ¹⁴C-naphthalene and ¹⁴C-benzo[a]pyrene was measured. An amino acid mixture was taken as reference substrate for total heterotrophic activity. The rate of uptake of naphthalene was found to increase with decreasing distance from the oil platform at all fields with maximum values at stations 0.5 miles (ca 0.8 km) from the centre of oil activity. Data on naphthalene uptake parallel GC-MS data for aromatic hydrocarbons and allow an estimation of the actual uptake velocity of naphthalene in the sediment to be made. A significant mineralisation of benzo[a]pyrene could only occasionally be detected. The respiration of amino acids did not show any pattern related to the oil field activities.     

Rate and extent of decomposition of dissolved organic matter in surface seawater

An experiment on the decomposition of the dissolved organic matter (DOM) in surface seawater was carried out under controlled laboratory conditions during a cruise of the R.V. Hakuho-Maru in the North-Equatorial Pacific (August/November, 1969). Surface seawater was placed in 300 ml oxygen bottles and incubated in the dark at 25°C. After selected time intervals, the dissolved organic-carbon (DOC) and the dissolved oxygen (DO) were determined for each bottle. The DOM in the surface seawater was described in terms of two labile fractions and a refractory one. The DOC decreased from 0.96 to 0.74 mgC/l during the first 50 days of incubation. Approximately 20% of the initial DOC (Fraction F_I) was oxidized. The other labile fraction, F_II was assumed to be 30 to 40% of the total DOM. The remaining half of the total DOM (Fraction F_III) is probably refractory and resistant to biochemical oxidation. The rate constant for decomposition was 0.0052 day^–1 for the total DOM and 0.033 day^–1 for F _I.     

Importance of refractory plant material to the carbon budget of the oyster Crassostrea virginica

The ability of the oyster Crassostrea virginica (Gmelin) to filter, ingest and assimilate ¹⁴C-labeled Spartina alterniflora as a carbon source was investigated under laboratory conditions. The oyster assimilated crude-fiber carbon extracted from S. alterniflora with an efficiency of approximately 3%. Enteric bacteria did not enhance this process. The annual average (April 1984 to November 1985) of crude fiber in the Choptank River sub-estuary of the Chesapeake Bay, Maryland, USA, from which the oysters were collected, was 15.7 g l^-1 (range 4.3 to 34.3 g l^-1). The potential food value of crude fiber to oysters in this system was estimated to be less than 1% of their carbon demand. However, the potential contribution of crude fiber to the carbon requirements of other oyster populations, such as those in southeastern USA, may be as great as 20%, due to higher crudefiber concentrations in the seston.     

Photosynthesis and amelioration of desiccation in the intertidal saccate alga Colpomenia peregrina

Rates of gross photosynthesis for the intertidal saccate alga Colpomenia peregrina (Sauv.) Hamel were determined under submersed and emersed conditions. Maximal photosynthetic rates were lower than for most seaweeds but comparable with other saccate members of the genus. By fitting the data to a hyperbolic tangent function, maximal photosynthetic rates were estimated to be 5.29 mmol CO₂ m^-2 h^-1 under submersed conditions and 2.06 mmol CO₂ m^-2 h^-1 under emersed conditions. I_k for submersed thalli was 69.1 E m^-2 s^-1, wherea for emersed thalli it was 149.0 E m^-2 s^-1, or 2.2 times higher. At low tide in the field and under saturating irradiance, carbon from seawater retained within the thallus cavity was assimilated at 0.9 mmol CO₂ m^-2 h^-1. In the laboratory under emersed conditions, carbon from this source was taken up at 0.6 mmol CO₂ m^-2 h^-1 at 20°C and at 0.34 mmol CO₂ m^-2 h^-1. Retained seawater also greatly reduced drying under desiccating conditions. Experimental thalli from which seawater had been removed lost thallus water continuously throughout the drying period (120 min). On the other hand, control, thalli lost water for the first 15 min, after which no further water loss occurred. At the termination of the experiment, control thalli had lost 7.2% of their water, whereas experimental thalli had lost 39.2%. Desiccation affected the emersed photosynthetic rate of experimental and control thalli. Emersed photosynthetic rates for thalli dried for 15 min were higher than for fullyhydrated thalli. However, emersed photosynthesis of thalli dried for longer than 15 min was lower than fully-hydrated rates and was directly related to percent water loss. Utilizing data from this study, a model was constructed to determine total photosynthetic production of C. peregrina over a single daylight period. From these calculations it was determined that emersed photosynthesis can increase daily photosynthetic production of C. peregrina by 50%.     

Acute toxicity of mercury and selenium to Crassostrea gigas embryos and Cancer magister larvae

The possible modification of mercury toxicity by selenium in embryos of the Pacific oyster Crassostrea gigas and the larvae of the crab Cancer magister was investigated. Mercury concentration eliciting abnormal development in 50% of the oyster embryos (EC₅₀) was 5.7 g l^-1 (48 h) and mortality in 50% of the crab larvae (LC₅₀) occurred with 6.6 g l^-1 (96 h). The 48 h EC₅₀ for selenium was greater than 10,000 g l^-1 for oyster embryos and the 96 h LC₅₀ for crab zoeae was 1040 g l^-1. The response from each species, when exposed to both toxicants, revealed, that a high level of selenium (5,000 g l^-1) increased mercury toxicity. Moderate selenium concentrations (10 to 1,000 g l^-1) tended to decrease mercury toxicity, although no statistical verification could be made. The order of administration of toxicants had no effect on the response of Crassostrea gigas embryos. Early developmental stages (8 h) of C. gigas embryos were most sensitive to dissolved Hg; toxicant administration 24 h after fertilization resulted in no apparent abnormalities in development.     

Temperature and salinity adaptation in the purple shore crab Hemigrapsus nudus: An in vitro metabolic flux study with excised gills

The effects of temperature and salinity acclimation on intermediary metabolism in excised gill homogenates from Hemigrapsus nudus were examined. In general, a decrease in salinity was followed by an increase in the oxidation of the substrates glucose-1-¹⁴C, acetate-1-¹⁴C and glycine-¹⁴C to ¹⁴CO₂. Also, there was an increase in amino acid incorporation into the protein fraction. Both of these metabolic parameters were differentially temperature-sensitive. An enzymic model which may explain the increase in respiration rate observed in several intertidal invertebrates meeting an hypo-osmotic stress is proposed, using data from these experiments and also from the literature.     

Allozyme variation in European populations of the oyster Ostrea edulis

Variations at 22 enzyme coding loci were surveyed in 11 populations of the oyster Ostrea edulis L., which were sampled between 1988 and 1990 along the Atlantic and Mediterranean coasts of Europe. Atlantic oyster beds suffered a steady decline during the last century, and restocking of beds with oysters of foreign origin has probably resulted in a high degree of interbreeding of natural oyster stocks from all Atlantic Europe. Our study confirms the low levels of genetic variability previously reported for the oyster populations from the Atlantic coasts, and extends it to the Mediterranean coasts. The locus arginine-kinase (ARK ^*) exhibited a high degree of interpopulation differentiation (F _ST=0.289), resulting from extensive variation in gene frequencies along a geographical cline. However, the overall genetic differentiation between populations was slight, and similar to that reported for other local populations of bivalves (mean genetic distance between populations is 0.010, mean F _ST=0.062). A general pattern of increasing differentiation along the coastline in an Atlantic-mediterranean direction emerged; but genetic differentiation among the Atlantic populations was not significantly lower than that observed among the Mediterranean populations. This and other results suggest that the effects of extensive transplantation of oysters among various areas in Europe are detectable only in some particular localities. The geographical distribution of low-frequency alleles suggests a restriction to gene flow outwards from the Mediterranean Sea, across the Straits of Gibraltar.