Sublethal effects of chronic oil exposure on the intertidal clam Macoma balthica

When exposed to Prudhoe Bay crude oil in flowing seawater for 180 days, the small intertidal clam Macoma balthica showed behavioral, physical, physiological and biochemical changes. At a high concentration of oil in seawater (3.0 mg l^-1) burrowing rate decreased, respiration rate increased, growth was inhibited, and very high mortalities resulted. The lowest concentration of oil in seawater (0.03 mg l^-1) inhibited growth and caused reabsorption of gametes. One group of adverse oil effects which was related to sluggishness and disorientation of the clams appeared after a week of exposure to oil; another group related to a negative energy balance was not observed until 60 days. We conclude that chronic exposure of M. balthica to oil-in-seawater concentrations even as low as 0.03 mg l^-1 will, in time, lead to population decreases.Please address requests for reprints to Dr. D. G. Shaw at the Institute of Marine Science     

Oxygen dynamics and transport in the Mediterranean sponge Aplysina aerophoba

The Mediterranean sponge Aplysina aerophoba kept in aquaria or cultivation tanks can stop pumping for several hours or even days. To investigate changes in the chemical microenvironments, we measured oxygen profiles over the surface and into the tissue of pumping and non-pumping A. aerophoba specimens with Clark-type oxygen microelectrodes (tip diameters 18–30 μm). Total oxygen consumption rates of whole sponges were measured in closed chambers. These rates were used to back-calculate the oxygen distribution in a finite-element model. Combining direct measurements with calculations of diffusive flux and modeling revealed that the tissue of non-pumping sponges turns anoxic within 15 min, with the exception of a 1 mm surface layer where oxygen intrudes due to molecular diffusion over the sponge surface. Molecular diffusion is the only transport mechanism for oxygen into non-pumping sponges, which allows total oxygen consumption rates of 6–12 μmol cm⁻³ sponge day⁻¹. Sponges of different sizes had similar diffusional uptake rates, which is explained by their similar surface/volume ratios. In pumping sponges, oxygen consumption rates were between 22 and 37 μmol cm⁻³ sponge day⁻¹, and the entire tissue was oxygenated. Combining different approaches of direct oxygen measurement in living sponges with a dynamic model, we can show that tissue anoxia is a direct function of the pumping behavior. The sponge-microbe system of A. aerophoba thus has the possibility to switch actively between aerobic and anaerobic metabolism by stopping the water flow for more than 15 min. These periods of anoxia will greatly influence physiological variety and activity of the sponge microbes. Detailed knowledge about the varying chemical microenvironments in sponges will help to develop protocols to cultivate sponge-associated microbial lineages and improve our understanding of the sponge-microbe-system.     

Growth of juvenile Mercenaria mercenaria and the effect of resuspended bottom sediments

The influence of silt on growth of juvenile hard clams Mercenaria mercenaria (L.) (9 mm in mean shell length) was investigated in the laboratory using mixed suspensions of algae (50x10⁶ Pseudoisochrysis paradoxa cells l^-1) and fine-grained bottom sediments (0 to 44 mg l^-1). Growth rates, expressed as percent increase in ash-free dry tissue weight, were not significantly affected by sediment concentrations up to 25 mg l^-1. Significant reduction in growth (by 16% relative to controls fed only algae), and condition of clams, occurred at 44 mg silt l^-1. The results of the 3-week growth experiment agree well with predictions made in an earlier study by integrating results of shortterm physiological measurements. Growth rates obtained with experimental algal-silt diets at 21°C (2.6 to 3.3% increase in dry tissue weight d^-1) were comparable to those determined at ambient concentrations of Great South Bay particulates at 20°C (0.9 to 4.0% d^-1). Levels of particulate inorganic matter in seawater from Great South Bay, New York, exhibited pronounced daily changes, and ranged from 6 to 126 mg dry weight l^-1. Growth enhancement by the addition of silt to an algal diet, reported in mussels, surf clams and oysters, was not found in M. mercenaria. It is suggested that these three species are better suited than hard clams for culturing efforts in inshore turbid waters above uncompacted, muddy bottoms.Contribution No. 452 from the Marine Sciences Research Center, State University of New York at Stony Brook, USA     

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     

Short-term effect of microencapsulated hydrocarbons on shell growth of Mytilus edulis

Two North Sea crude oils, weathered crude oil, n-alkanes, medical liquid paraffin and a fish oil were microencapsulated and added to seawater. This gave a medium containing both water soluble fractions (WSF) and oil particles, and thus simulated the conditions in natural seawater contaminated with oil. The microencapsulated oils were fed to Mytilus edulis L. in different concentrations, and the growth in terms of shell length of the mussels was measured in intervals of 24 to 48 h for 4 to 12d. With the crude oilss A and B, weathered crude oil and n-alkanes at concentrations1 mg l^-1, shell growth rate decreased rapidly compared to controls. With liquid paraffin at levels of 1 to 12 mg l^-1, a small but significant negative effect on shell growth occurred after 5 d of exposure. Exposure to fish oil at 1 and 4 mg l^-1 gave no significant reduction in growth rate. The toxicity of the different hydrocarbons was not related to their content of aromatic fractions. Crude oil B was tested at concentrations ranging from 0.12 to 12 mg l^-1. At 0.12 mg l^-1 shell growth was not significantly different from the control, while at 0.25 and 0.50 mg l^-1 a temporary and significant stimulation of growth was observed. The product concentration (C)xresponse (R) shows a linear regression on exposure time (t). The regression model CxR=75-0.18t is used to estimate EC-values (effect of a given concentration) for given response levels. The model gives a very good fit to observed data.     

Respiratory and behavioral responses of the grass shrimp Palaemonetes pugio to cadmium and reduced dissolved oxygen

Adult grass shrimp (Palaemonetes pugio) were collected during July and August, 1973, and exposed for 48-h periods to a series of sublethal cadmium (Cd) concentrations: 0.1 and 0.56 mg l^-1 for metabolic studies; and 0.01, 0.05, 0.1, 0.3, 0.56 mg l^-1 for locomotor studies. Treatment effects were tested singly and in combination at selected dissolved oxygen (DO) concentrations ranging from near saturation to 0.8 mgl^-1. Shrimp not exposed to Cd exhibited metabolic regulation between 6.5 and, 4.6 mg DOl^-1, with progressively lower O₂ consumption rates at lower DO concentrations. Oxygen consumption of Cd-exposed shrimp was suppressed overall relative to the non-Cd treated shrimp, and with one exception, the magnitude of metabolic suppression directly corresponded to Cd concentration. Locomotor activity of non-Cd exposed shrimp was relatively independent of DO concentration down to 2.8 mg DO l^-1 and became significantly suppressed at 1.8 mg DO l^-1 and below. Significant suppression of locomotion was evident in shrimp exposed to 0.56 mg Cd l^-1, but was not seen in treatment groups of 0.3 mg Cd l^-1 or lower. No interaction on locomotor activity of shrimp was detected between Cd exposure and oxygen concentration. The reduction of locomotor activity and metabolism at 0.56 mg Cd l^-1 exposure is similar to that occurring under reduced oxygen condition. It is hypothesized that Cd acts in some manner to suppress O₂ availability to P. pugio, a suggestion consistent with literature reports on adverse effects of Cd on fish respiratory enzymes and gill tissue.Contribution No. 351 of the U.S. Environmental Protection Agency, Environmental Research Laboratory, Narragansett, Rhode Island. Although the research described in this article was funded by the U.S. Environmental Protection Agency, it has not been subjected to Agency review and therefore does not necessarily reflect the views of the Agency and no official endorsement should be inferred     

Influence of algal and suspended sediment concentrations on the feeding physiology of the hard clam Mercenaria mercenaria

Short-term laboratory feeding experiments were conducted to determine the response of the hard clam Mercenaria mercenaria (L.) (32 mm in mean shell length) to increasing sediment concentrations. Clams were fed mixed suspensions of Pseudoisochrysis paradoxa (50 and 150 cells l^-1) and bottom sediments (0 to 44 mg l^-1). Algal ingestion rate deelined with increasing sediment loads. This resulted primarily from a reduction in clearance rate, which declined by 0.08 l h^-1 g^-1 (1.3%) for every 1 mg l^-1 increase in sediment load. This reduction was of similar magnitude for juvenile (13 mm) clams. At the algal concentrations tested, pseudofaeces production was intermittent and inconspicuous below about 10 mg silt l^-1. Loss of algae in pseudofaeces increased with increasing sediment loads; however, even at the highest silt and algal concentrations, clams lost a maximum of only 18% of the algae cleared from suspension. Thus, pseudofaeces production is not expected to cause significant loss of algal food at the sediment concentrations normally encountered in the natural environment ( ca 40 mg silt l^-1). Absorption rate of total organic matter remained constant, at least up to silt concentrations of 20 mg l^-1. Experiments using dual ⁵¹Cr:¹⁴C-formaldehyde-labelled sediment indicated that clams were able to counteract the dilution of algae by absorbing a considerable fraction (21 to 22%) of detrital sedimentary organics. Absorption efficiency of pure P. paradoxa ranged from 82% at 50 cells l^-1 to 58% at 150 cells l^-1. Integration of physiological rate measurements suggests that at moderate to high algal concentrations (300 g Cl^-1), growth improvement by the addition of silt, documented in mussels, surf clams and oysters, is unlikely to occur in M. mercenaria. It is suggested that a suspension-feeding bivalve's success in maximizing its energy gain in a turbid environment depends on the combination of two features: a high selection efficiency and a high rate of pseudofaeces production. It is proposed that species which regulate ingestion primarily by producing pseudofaeces are better adapted to cope with high suspended sediment loads than species such as M. mercenaria, which control ingestion mainly by reducing clearance rate.Contribution No. 451 from the Marine Sciences Research Center, State University of New York at Stony Brook, USA     

Fluxes of dissolved and particulate calcium in selected tissues of Crassostrea virginica

Rates of calcium incorporation by selected tissues of Crassostrea virginica increased in a step-wise fashion from lowest values among organisms exposed to ambient total calcium concentrations of 45 mg l^-1, intermediate values among oysters exposed to 135, 225, and 315 mg l^-1, to highest rates for oysters exposed to 360 mg l^-1. Although excised visceral mass tissue had highest rates of calcium incorporation relative to mantle, muscle, and both organic and inorganic portions of the shell, mantle tissue appeared to have the most dynamic response to changes in ambient calcium concentrations. Rates of dissolved calcium incorporation from ambient water were approximately two to three orders of magnitude higher than comparable rates from ingested algal food. Behavioral response to concentrations of selected inoic, species in the ambient environment may have been responsible for observed differences in rates of calcium incorporation.     

Effect of copper on ion- and osmoregulation in the shore crab Carcinus maenas

The uptake and effect of dissolved copper on regulation of hemolymph osmolality and Na⁺, K⁺, Cl^-, Ca⁺⁺, and Mg⁺⁺ concentrations in the shore crab Carcinus maenas (L.) were determined at 400 mOsm (=14 S) ambient salinity. One mg Cu l^-1 resulted in 50% mortality in 11 to 22 d; the highest sensitivity was observed around the moulting period. 0.25 and 0.5 mg Cu l^-1 were not lethal after a onemonth exposure. Ten, 1, and 0.5 mg Cu l^-1 altered regulation of osmolality, Na⁺, K⁺, and Cl^- concentrations, while 0.25 mg Cu l^-1 did not. Exposure to 1 mg Cu l^-1 reduced hemolymph osmolality and Na⁺, K⁺, and Cl^- concentrations to 80 to 90% of controls within 4 to 6 d and no further reduction was observed during a 21-d exposure. The effect of various copper concentrations on these four parameters were almost identical and the highest sensitivity was observed around the moulting period. Hemolymph calcium levels increased 20 to 80% in crabs exposed to 1 mg Cu l^-1, while they decreased 20 to 30% in crabs exposed to 0.5 mg Cu l^-1. Prolonged exposure to copper caused 20 to 70% reductions in hemolymph magnesium levels. Crabs exposed to 0.5 mg Cu l^-1 for 29 d accumulated copper in hepatopancreas, gills, carapace, heart, testes, and hypodermis, but not in muscles and hemolymph. Increased copper levels in crabs exposed to 0.25 mg Cu l^-1 were observed in hepatopancreas, gills, and carapace only. The present results suggest that effects of copper on ion regulatory processes in part explain why the toxicity of copper towards euryhaline invertebrates increases at low salinities.     

Relationship between lead concentrations in seawater and in the mussel Mytilus edulis: a water-quality criterion

Regression analysis on data collected from Port Phillip Bay and Western Port, Australia in 1979 shows that there is a significant equilibrium relationship between total recoverable lead in seawater and its concentrations in the mussel Mytilus edulis (P<0.001). The concentration of lead in seawater should not exceed 1.27 g l^-1 if the mussel is not to reach a lead concentration of 2.5 mg kg^-1 wet weight, a value frequently used as a food standard for human consumption. When a lead value of 2.5 mg kg^-1 wet weight is reached, the concentration factor by mussels for lead from seawater is 1969. The critical value of 1.27 g l^-1 could be used as a marine water-quality criterion for lead in waters where mussels are harvested.     

Respiration,pumping activity and heart rate in Ciona intestinalis exposed to fluctuating salinities

Specimens of Ciona intestinalis L. were exposed to both gradual (sinusoidal) and abrupt (square-wave) salinity fluctuations and the changes in pumping activity and oxygen sonsumption monitored. Heart rate was monitored under steady state conditions using a new in vivo method, and pumping activity was monitored as spontaneous squirting by use of a pressure transducer. Oxygen consumption was also monitored under steady state conditions and under conditions of declining oxygen tension. It was found that during periods of regular beating the mean heart rate for 5 ascidians was 13.2 beats min^-1 for the branchial pacemaker and 32.6 beats min^-1 for the visceral pacemaker. Regular periods of beating lasted for approximately 1 min, followed by periods of irregular activity prior to heart beat reversal. The heart beat became irregular and reversal occurred sporadically when the ascidians were exposed to dilute seawater. The ascidians showed a pumping rate of 14 squirts per hour in full-strength seawater. This rate declined during decreasing salinity and ceased entirely when the external seawater concentration reached approximately 60% seawater (100%=32% S). During periods of decreased salinity, the siphons were tightly closed and oxygen consumption was zero. The rate of oxygen consumption by C. intestinalis decreased during decreasing salinities and there was no evidence of an oxygen debt. Oxygen consumption under steady state conditions varied with body weight according to the following: oxygen consuption =0.515 W ^0.831 (where W is body weight). The rate of oxygen consumption was found to be dependent on the external oxygen tension.     

N-acyl homoserine lactone production by bacteria within the sponge Suberites domuncula (Olivi, 1792) (Porifera, Demospongiae)

Many bacteria live in close association with sponges. Within these consortia, molecules of communication such as quorum-sensing and hormone-like molecules may occur in order to regulate the partnership. Of particular interest, bacterial N-acyl-l-homoserine lactones (AHLs) were screened in supernatants from Suberites domuncula-associated bacteria using an E. coli bioluminescent reporter system. These sponge-associated bacteria were beforehand isolated on several media supplemented or not with a sponge extract to attempt to isolate sponge-specific bacteria. Out of 81 AHL-producing bacteria, three strains requiring sponge extract to grow were selected for AHL characterization. The in vitro produced AHLs, that is, in bacterial culture supernatants, were identified as N-(3-butanoyl)-l-homoserine lactone and N-(3-oxododecanoyl)-l-homoserine lactone and quantified using LC–ESI–MS/MS. The in vivo production of AHLs by sponge-associated bacteria has also been demonstrated in a healthy host for the first time: N-(3-oxododecanoyl)-l-homoserine lactone, N-(3-hexanoyl)-l-homoserine lactone, and N-(3-heptanoyl)-l-homoserine lactone. This AHL production in sponges may suggest a potential role of these molecules between sponge-associated bacteria and/or between sponge-associated bacteria and the sponge.     

Redwood of the reef: growth and age of the giant barrel sponge Xestospongia muta in the Florida Keys

The growth of animals in most taxa has long been well described, but the phylum Porifera has remained a notable exception. The giant barrel sponge Xestospongia muta dominates Caribbean coral reef communities, where it is an important spatial competitor, increases habitat complexity, and filters seawater. It has been called the ‘redwood of the reef’ because of its size (often >1 m height and diameter) and presumed long life, but very little is known about its demography. Since 1997, we have established and monitored 12 permanent 16 m diameter circular transects on the reef slope off Key Largo, Florida, to study this important species. Over a 4.5-year interval, we measured the volume of 104 tagged sponges using digital images to determine growth rates of X. muta. Five models were fit to the cubed root of initial and final volume estimates to determine which best described growth. Additional measurements of 33 sponges were taken over 6-month intervals to examine the relationship between the spongocoel, or inner-osculum space, and sponge size, and to examine short-term growth dynamics. Sponge volumes ranged from 24.05 to 80,281.67 cm³. Growth was variable, and specific growth rates decreased with increasing sponge size. The mean specific growth rate was 0.52 ± 0.65 year⁻¹, but sponges grew as fast or slow as 404 or 2% year⁻¹. Negative growth rates occurred over short temporal scales and growth varied seasonally, significantly faster during the summer. No differences in specific growth rate were found between transects at three different depths (15, 20, 30 m) or at two different reef sites. Spongocoel volume was positively allometric with increasing sponge size and scaling between the vertical and horizontal dimensions of the sponge indicated that morphology changes from a frustum of a cone to cylindrical as volume increases. Growth of X. muta was best described by the general von Bertalanffy and Tanaka growth curves. The largest sponge within our transects (1.23 × 0.98 m height × diameter) was estimated to be 127 years old. Although age extrapolations for very large sponges are subject to more error, the largest sponges on Caribbean reefs may be in excess of 2,300 years, placing X. muta among the longest-lived animals on earth.     

Accumulation,fractionation and release of oil by the intertidal clam Macoma balthica

The bivalve mollusc Macoma balthica accumulated hydrocarbons during 180 days of continuous exposure to Prudhoe Bay crude oil in seawater dispersions with nominal concentrations of 0.03, 0.3 and 3.0 mg l^-1. The mollusc's ability to concentrate oil from seawater increased with decreasing oil-in-water concentration. Decreases in oil burden began after 30 to 120 days (depending on the oil concentration) and continued for at least 60 days after exposure to oil ceased. Aliphatic and aromatic hydrocarbons were fractionated in markedly different ways by the bivalve. Branched and cyclic aliphatics in the molecular weight-range dodecane through hexadecane were preferentially retained over straightchain and their higher homologs. Larger and more substituted aromatic compounds were selectively concentrated. There appeared to be no selective concentration of aromatic sulphur compounds.Please address requests for reprints to Dr. D. G. Shaw at the Institute of Marine Science     

An improved automatic recording apparatus for determining the filtration rate of Mytilus edulis as a function of size and algal concentration

An automatic recording apparatus for measuring the filtration rate in suspension-feeding bivalves is described. The concentration of algae in the experimental medium is kept constant throughout each experiment by addition of Phaeodactylum tricornutum from a chemostat. Within the range of body size 5.7 to 283 mg (W=dry weight of tissues), the filtration rate (F=ml min^-1) at 15°C in Mytilus edulis L. follows the allometric equation F=0.85 W ^0.72. Within the concentrations 0.18 to 0.70 mg algal dry weight l^-1, the filtration rate in mussels of 132 mg dry flesh weight ranges from 33.1 to 41.0 ml min^-1. At 0.18 mg algal dry weight l^-1 the mussels filter continuously for 20 h, with a high constant rate that presumably represents the water transport capacity under optimal laboratory conditions.     

Observations of the effect of zinc on the gametes and various development phases of Arbacia lixula

The action of various concentrations of zinc on different phases of development of Arbacia lixula L. has been observed. The following concentrations were used in the experiment: 0.01, 0.1, 1 and 10 mg l^-1; these concentrations were tested on unfertilized eggs, sperms, fertilized eggs and adult individuals. It was found that after 96 h a 0.01 mg l^-1 concentration reduces the mobile sperm percentage compared with the control group. At concentrations of 0.1, 1 and 10 mg l^-1 no mobile sperm were found after 96 h. Compared with the control group, the action of zinc on unfertilized eggs was to increase the percentage of eggs showing signs of cytolisis after 12, 36 and 60 h. In the case of fertilized eggs it was found that with a concentration of 0.01 mg l^-1 the P₅ pluteus stage was attainable; the percentage of forms reaching that stage was less than in the control group. At 0.1 mg l^-1 only a very low percentage of eggs could reach the P₅ pluteus stage; concentrations of 1 and 10 mg l^-1 are incompatible with the normal development of an egg. Adult individuals kept in aquaria with 1 mg l^-1 zinc showed no signs of suffering after 20 d. Gametes are proposed as the appropriate material for carrying out bioassays.     

Relationship between cadmium concentrations in seawater and those in the mussel Mytilus edulis

Various forms of regression analysis are presented which show that there is a significant equilibrium relationship between total recoverable cadmium in seawater and its concentration in the mussel Mytilus edulis (P<0.001). The calculations show that the concentration of cadmium in seawater should not exceed 0.20 g l^-1 if the mussel is not to reach a cadmium concentration of 2 mg kg^-1 wet weight: a value frequently used as a food standard for human consumption. Further, when a cadmium value of 2 mg kg^-1 wet weight is reached, the concentration factor by mussels for cadmium from seawater is 9 950.     

Effect of uranium on growth and reproduction of the marine amphipod Allorchestes compressa

Experiments on growth of the marine amphipod Allorchestes compressa Dana were carried out over four weeks, and both growth and reproduction were studied over three generations, each of which was exposed to uranium for approximately 10 wk. At 0.1 mg l^-1 the uranium increased growth by 23%, as measured by the mean weight after 4 wk, and at 2 mg l^-1 growth was reduced by 28% compared with the control. A. compressa accumulated uranium from sea water with a concentration factor of 10. There was no effect of uranium on the survival of amphipods or their progeny in the multiple-generation experiment, but the numbers of males, the sex ratio, and the respiration rate (measured on males only) at 1mg l^-1 were significantly lower than the control. A. compressa is shown to be a convenient species for the study of toxic effects on growth and reproduction using multiple-generation experiments.     

Silica uptake kinetics of Halichondria panicea in Kiel Bight

Besides diatoms Demospongiae are the most important consumers of dissolved silica in the sea. They can play an important role for the silica budget especially in the shallow water areas of the Baltic Sea. The dependence of the silica uptake rate on the silica concentration of the seawater was measured for the sponge Halichondria panicea (Pallas, 1766). The sponges were collected in Kiel Bight. The uptake conformed to Michaelis–Menten kinetics with a half-saturation constant of 46.41 μM and a saturated uptake rate of 19.33 μmol h⁻¹ g⁻¹ ( p < 0.01). In the red algae zone of Kiel Bight the sponges depend on silica supply from the surrounding waters and may be silica-limited rather than food-limited in growth. Because of the much faster uptake of silica by diatoms and their lower saturation point, as well as the difference in spatial distribution of the two main silica consumers, a competition for silica between sponges and diatoms seems unlikely. Received: 21 June 1997 / Accepted: 15 July 1997     

The effect of low concentrations of phenol and ammonia on egg production rates,fecal pellet production and egg viability of the calanoid copepod Acartia clausi

Estimates of daily fecundity, hatching success and fecal pellet production are reported for Acartia clausi females exposed for 10 d to low levels of phenol and ammonia. Copepods were collected in 1991 and 1992 from a southern coastal area of the Mar Grande of Taranto (southern Italy). A reduction in egg numbers and fecal pellet production was observed for females after 8 d of exposure to 500 g l^-1 phenol concentration. Ammonia (120 g l^-1) produced a significant increment in egg production, but hatching success was reduced by about 50% after nine exposure days. A. clausi was more sensitive to ammonia than phenol at high concentrations (24-h LC₅₀ phenol-32.26 mg l^-1; 24-h LC₅₀ ammonia=0.91 mg l^-1). At low concentrations, only long-term exposure to phenol determined a reduction in fecundity.