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.     

Bioenergetics and survival of the marine snail Thais lima during long-term oil exposure

The carnivorous snail Thais lima was fed Mytilus edulis during a 28-d exposure to the water soluble fraction (WSF) of Cook Inlet crude oil. The LC-50 of T. lima declined from >3000 ppb aromatic hydrocarbons on Day 7 to 818±118 ppb on Day 28. The LC-50 of M. edulis declined from >3 000 ppb aromatic hydrocarbons on Day 7 to 1 686±42 ppb on Day 28. Predation rate declined linearly with increasing aromatic hydrocarbon concentration up to 302 ppb; little predation occurred at 538 ppb and none at 1 160 or 1 761 ppb. Snail absorption efficiency averaged 93.5% and did not vary as a function of WSF dose. Total energy expenditure (R+U) increased at 44 ppb aromatics and declined at lethal WSF exposures. At sublethal WSF exposures, percentages of total energy expenditure were: respiration (87%), ammonia excretion (9%) and primary amine loss (4%). These percentages did not vary as a function of WSF dose or time. Oxygen:nitrogen ratios were not affected by WSF concentration or time and indicated that T. lima derived most of its energy from protein catabolism. The uptake of aromatic hydrocarbons into the soft tissues of snails and mussels was directly related to the WSF concentration. Naphthalenes accounted for 67 to 78% of the aromatic hydrocarbons in T. lima and 56 to 71% in M. edulis. The scope for growth was negative above 150 ppb WSF aromatic hydrocarbons and above 1 204 ppb soft-body aromatic hydrocarbons. These snails were physiologically stressed at an aromatic hydrocarbon concentration which was 19% of the 28-d WSF LC-50 (818±118 ppb) and/or 48% of the 28-d LC-50 of soft tissue aromatics (2 502 ppb).     

Formation of metallothioneins in relation to accumulation of cadmium in the common mussel Mytilus edulis

Accumulation rates of Cd have been studied in the common mussel Mytilus edulis L. under different laboratory conditions. Semi-static and through-flow experiments were carried out at Cd concentrations of 200 ppb and 50/100 ppb in the water, respectively. A linear uptake of Cd throughout the experimental periods of 36 or 124 d was found in all experiments. The accumulation rate was 3.10 ppm d^-1 at 50 ppb in the through-flow water experiment, 4.1 ppm d^-1 in starved and 6.6 ppm d^-1 in fed mussels in the semi-static experiments with 200 ppb Cd in the water. The Cd-accumulation rate in various fractions of soft parts decreased in the order: body>mantle>muscles. Elution patterns obtained from gel-filtrations showed an increasing amount of Cd-binding proteins (metallothioneins) when the body burden of Cd increased. The Cd content in the body fraction of mussels transferred to clean water for 42 d after first being exposed to Cd during 124 d decreased from 564 ppm on a dry weight basis to 417 ppm while the fraction of Cd bound to metallothioneins rose from 22 to 78% during the same period.     

Chemical composition and effects of water extracts of petroleum on eggs of the sand dollar Melitta quinquiesperforata

Sperm and eggs of sand dollars, Melitta quinquiesperforata (Leske), were subjected to two petroleum oils, and effects determined. The oils chosen were Kuwait crude and No. 2 fuel oil, supplied by the American Petroleum Institute. Water-soluble extracts (WSF) from oil-sea water mixes were prepared and the major aromatic components in the WSF of the fuel oil were identified. WSF of No. 2 fuel oil depressed respiration, mobility of sperm, interfered with fertilization and cleavage, and retarded larval development. The effects were detectable at dilutions of 4% and less (about 0.6 ppm of WSF). Kuwait crude was much less toxic. There was no effect on water permeability of the egg membrane. Results are compared with similar studies on other marine organisms.     

Mussels flexing their muscles: a new method for quantifying bivalve behaviour

We employed a novel technique to quantify how blue mussels Mytilus edulis react to predation risk in their environment by quantifying mussel gape using a Hall sensor attached to one shell valve reacting to a magnet attached to the other. Change in gape angle per second (CHIGA) versus gape angle plots resulted in a distribution with a boundary, which defined the maximum CHIGA of a mussel at all gape angles. CHIGA boundary plots for all individual mussels were similar in form. However, the CHIGA boundary increased in extent with mussel length (maximum CHIGA for mussel valve closures for mussels 2.98 and 79.6 mm long were −1.5 and −11°s⁻¹, respectively), showing that larger mussels opened and closed most rapidly. Mussel extract added to the seawater, a factor believed to signal predation, caused mussels to close significantly faster than otherwise (P < 0.001). This approach for assessing how mussels react to their environment indicates that mussel response to predation is graded and complex and may well indicate animal-based assessments of the trade-off between effective feeding and the likelihood of predation.     

A long-term study of the effects of water-soluble fractions of No. 2 fuel oil on the survival,development rate,and growth of the mud crab Rhithropanopeus harrisii

Mud crabs, Rhithropanopeus harrisii (Gould), were exposed continuously for 6 months after hatching to water-soluble fractions (WSF) of No. 2 fuel oil. Survival, growth and development rate were monitored during this time. The zoeal stages were the most sensitive to fuel oil. A 20% WSF (0.36 ppm total naphthalenes, 1.26 ppm total hydrocarbons) was acutely toxic to these stages. Of the zoeal stages, the first stage appeared to be the most sensitive. The combined duration of the 4 zoeal stages was significantly increased by increasing WSF exposure concentrations. The megalopa and crab stages were not particularly sensitive to continued petroleum hydrocarbon exposure, particularly when compared to zoeal stages. However, mean duration of the megalopa and first crab stages was significantly affected by oil exposure. Individuals which survived the highest exposure concentrations as larvae appeared to grow larger during the crab stages, so that at the end of 6 months comparably staged crabs were equal to or larger than both control crabs and those exposed to low WSF concentrations. Stage distributions at the end of 6 months showed no differences due to WSF exposure. Sex ratios, which could be determined at the end of 6 months, were approximately 1, indicating no sex-related differential sensitivity to WSF exposure, at least as larvae or juveniles. The data indicate that these crabs possess considerable ability to recover from the effects of chronic sublethal exposure to petroleum hydrocarbons. The most deleterious effects of oil pollution on this species may be due to its impact on larval recruitment into the adult population.     

Light and electronmicroscopic studies of the acute and long-term toxic effects of N-nitrosodipropylamine and N-methylnitrosurea on the marine mussel Mytilus edulis

The acute (1) and long-term (2) toxic effects of N-nitrosodipropylamine (NDPA) and N-methylnitrosurea (NMU) were studied by light and electronmicroscopy following injection of the chemicals into the foot of the mussels Mytilus edulis (L.) (1) Mussels reveived one injection of NDPA at concentrations of 0.25, 0.5, 1.0, and 2.0 mg NDPA mussel^-1, and one injection of NMU at concentrations of 0.25, 0.5, 1.0, and 4.0 mg NMU mussel^-1. Within 12d, necrosis of the epithelial lining of the digestive diverticula took place, and the vesicular connective tissue in the digestive diverticula were heavily infiltrated by eosinophilic granular leucocytes. (2) Mussels received four weekly injections of 0.5 mg NDPA mussel^-1, and 0.5 mg NMU mussel^-1. NDPA-exposed mussels were examined for a period of 17 weeks, and NMU-exposed mussels were examined for a period of 24 weeks. Both chemicals produced progressive tissue damage, characterized by destruction of digestive tubules and ducts in the digestive diverticula. Destroyed tubules and ducts were replaced by collagenous scars, and granulocytomas were frequently seen.     

Effects of fertilization and development of the common mussel Mytilus edulis after long-term exposure to a nonionic surfactant

Common mussels (Mytilus edulis L.) were exposed over a 5-month period to low-level concentrations (0.5 to 1.5 ppm) of a nonionic surfactant. Upon maturation at the end of this period, spawning ability was examined. Fertilization occurred at low-level concentrations (0.1 to 2.0 ppm) of the surfactant, and was most successful for gametes from the long-term controls and the highest long-term concentration (1.5 ppm). Inhibited or delayed larval development was observed, related to the concentration gradient of the short-term exposures. Gametes from mussels long-term exposed to the surfactant were more sensitive than those from the long-term control.     

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.     

Metallothionein induction inMytilus edulis exposed to cadmium

The exposure of mussels,Mytilus edulis, collected from Whitsand Bay, southwest England, in August 1988, to sublethal concentrations of cadmium (400µg l^–1) for 65 d resulted in the induction of metallothionein (MT) synthesis in the soft tissues. In cadmium-exposed mussels, metallothionein concentrations, measured by differential pulse polarography, increased by a factor of three, from 2 to 3 mg g^–1 to a maximum of 9 mg g^–1 after 30 d. No significant changes could be detected in controls. Cadmium accumulated in the soft tissues of mussels correlated significantly with metallothionein concentrations and can be described by the relationship: MT (mg g^–1)=0.045 Cd (µg g^–1)+3.03 (r=0.803,P<0.001). Gel chromatography of heat-treated cytosolic extracts showed that the accumulated cadmium is bound principally to the newly formed metallothioneins. Copper and zinc were also analysed in the whole soft-tissues and in subcellular fractions of cadmium-exposed mussels. Although copper concentrations were not affected by cadmium-exposure, zinc levels were significantly reduced. The results demonstrate that the induction of metallothioneins inM. edulis is a quantifiable biological response to sublethal levels of cadmium exposure.     

The musselsMytilus galloprovincialis andM. trossulus on the Pacific coast of North America

Most recent authors have called the bay mussels of the Pacific coast of North AmericaMytilus edulis Linnaeus, 1758. Thirteen samples ofedulis-like mussels were collected from California, Oregon, and Alaska, USA, in 1985, 1986 and 1987. Electrophoretic evidence from wight loci indicates that these samples consist of two genetically distinct groups, neither of which is similar toM. edulis from the Atlantic Ocean. Mussels in southern California are very similar toM. galloprovincialis Lamarck, 1819 from the Mediterranean Sea; it is probable thatM. galloprovincialis was introduced accidentally to southern California. Mussels in Oregon and Alaska are similar to those from the Baltic Sea and parts of eastern Canada; the nameM. trossulus Gould, 1850 has priority for this taxon. In central and nothern California,M. galloprovincialis, M. trossulus and their hybrids co-occur. Despite the presence of hybrids betweenM. galloprovincialis andM. trossulus, the genetic integrity which they maintain across large areas of the world warrants their recognition as two distinct species.     

Very long-chain aliphatic hydrocarbons in lipids of mussels (Mytilus edulis) suspended in the water column near petroleum operations off Sable Island,Nova Scotia,Canada

Mussels (Mytilus edulis) suspended in the water column in 1994 and 1995 for the monitoring of oil drilling operations off Sable Island, Nova Scotia were examined for hydrocarbon profiles, particularly aliphatic hydrocarbons. A spring bloom of phytoplankton occurred during the 90-d suspension period in 1995. Hydrocarbons isolated from the 1995 suspended mussels showed very high concentrations of both biogenic hydrocarbons and very long-chain n-alkanes from C₂₀ to C₃₂, initially thought to be petrogenic. Both types of hydrocarbons were either not detected or were only present in trace amounts in the mussels suspended in 1994 at similar sites. The biogenic hydrocarbons in the 1995 mussels were apparently of planktonic origin, from the spring bloom, and were dominated by heneicosahexaene (21:6), followed by pristance, heptadecane, and varions monounsaturated and polyunsaturated phytenes, heptadecenes, nonadecenes and heneicosenes. They could be readily hydrogenated to yield the basic alkanes. The 1995 mussels suspended within 1 km from the oil well platform were probably slightly tainted by petrogenic hydrocarbons, as evidenced by the detection of phytane and high concentrations of total aliphatic hydrocarbons, whereas the mussels suspended 10 km from the platform showed only high concentrations of biogenic hydrocarbons and the novel long-chain n-alkanes. The occurrence of an unusual phytoplankton bloom during the suspension period severely interfered with the petroleum monitoring role of mussels by altering the mussel hydrocrbon profiles through the accumulation into and probably selective depuration of xenobiotic hydrocarbons from the mussel, tissues.J. Parsons (deceased)     

Effects of oil pollution on the freezing tolerance and solute concentration of the blue mussel Mytilus edulis

Exposure of blue mussels (Mytilus edulis L.) to oil-polluted sea water for 7 d in the laboratory did not affect their body fluid concentrations of inorganic ions and free amino acids. Mussels exposed to-4°C for 12 h did not freeze, whereas freezing occurred invariably in mussels exposed to-7°C or lower temperatures for the same period. Following freezing at-10°C, oil polluted mussels resumed normal activity considerably more slowly than unpolluted ones, but oil-polluted as well as unpolluted mussels showed normal activity 3 h after thawing. Freezing at-15°C was invariably lethal to individuals of both groups. One possible explanation of the delayed recovery of oil-polluted mussels frozen at-10°C may be that oil components had become concentrated to toxic levels as the amount of solvent water diminished during freezing.     

Energy budgest,growth and filtration rates in Mytilus edulis at different algal concentrations

Growth of Mytilus edulis L. was measured in aquaria with through-flowing sea water at different levels of constant algal concentrations. The amount of food and oxygen consumed by the mussels were measured over given periods as well as the changes in dry organic weight during the same periods. From these parameters it was possible to make simple energy budgets and to compare the estimated growth with actual growth, and, further, to determine growth efficiences at different food levels. Energy budgets were made for mussels grown at algal concentrations of 0, 1.6×10³, 3.0×10³ and 26.0×10³ Phaeodactylum tricornutum cells x ml^-1. The estimated growth was found to be close to actual growth at algal concentrations above maintenance level and the net growth efficiency was found to be between 18% (3.0×10³ cells x ml^-1) and 61% (26×10³ cells x ml^-1). It has been shown that the filtration rate is independent of algal concentrations between about 1.5×10³ to 30×10³ P. tricornutum cells x ml^-1. Outside this range a decrease in filtration rate was noticed.     

Assimilation of trace elements by the mussel Mytilus edulis: effects of diatom chemical composition

Mussels have been widely used as bioindicators of coastal contamination, and recent reports have demonstrated that metals are accumulated from both the dissolved phase and from ingested food. In the winter and spring of 1995, we examined the influence of the chemical composition of food (protein content, trace element concentrations and ratios in the diatom Thalassiosira pseudomana) on the assimilation of six trace elements (Ag, Am, Cd, Co, Se and Zn) in the mussel Mytilus edulis (L.). Differences of up to 38% in diatom protein content had no major influence on the assimilation of any trace element or carbon. Protein assimilation in M. edulis examined with a ³⁵S radiotracer was also independent of protein content in the diatoms. Similarly, Se assimilation in mussels was not affected by the different Se concentrations in the diatoms. Cd assimilation increased with increasing Cd concentration, presumably due to higher desorption of Cd under acidic conditions typical of the mussel gut. Zn assimilation was inversely related to Zn concentration in the food particles, implying a partial regulation of this metal in the mussels. There was no evidence of any interaction of Cd and Zn in their assimilation by the mussels. These results suggest that mussels are highly responsive, in an element-specific way, to some components of ingested food (e.g., metal concentration), but other food components (such as the biochemical composition of the algae) have little effect on assimilation.     

Effect of Cd on accumulation and loss of 210Po in the mussel Mytilus edulis

As a result of ²¹⁰Po's previously identified association with sulphur-rich proteins, metallothioneins could have a significant effect on the behaviour and fate of ²¹⁰Po in molluscs. Starved control and cadmium-exposed mussels, Mytilus edulis, were fed ²¹⁰Po-labelled algae (Isochrysis galbana) for 5 d and then allowed to depurate in clean sea water. Cadmium-exposed M. edulis accumulated less ²¹⁰Po in the digestive gland and the remainder of the tissue than control mussels, although this was due to a decrease in tissue weight. More than 40% of ²¹⁰Po was identified as being associated with high molecular weight and heat-treated cytosol proteins in M. edulis. Mussels in a starved state are known to recycle as much as 90% of their amino acids. It is proposed that ²¹⁰Po associated with these and other proteins is recycled, explaining why no significant loss of ²¹⁰Po was observed from the remainder of the tissue in either control or Cd-exposed mussels. Cadmium-induced metallothioneins had no effect on the distribution of ²¹⁰Po in M. edulis; <5% associated with the cytosolic fraction was considered to principally contain metallothioneins. It is suggested that ²¹⁰Po's apparent relationship with metallothioneins is coincidental rather than connected with its role in the regulation of metals. Received: 30 March 1998 / Accepted: 3 August 1998     

Decrease of net carbon flux in two species of mussels caused by extracts of crude oil

Effects of crude oil and salinity stress on the metabolism of two common filter-feeding animals (Mytilus edulis and Modiolus demissus) have been investigated. Carbon budgets have been calculated for each species under a variety of combinations of oil content and salinity. Both reduced salinity and the presence of crude oil tend to decrease the net carbon flux for each species; stresses from each source interacted in their effects on experimental animals. Although similar responses to oil were shown by each species, Mytilus edulis appeared to be slightly more resistant to oil than Modiolus demissus.     

Geographic variation,niche breadth and genetic differentiation at different geographic scales in the mussels Mytilus californianus and M. edulis

Genetic differentiation was investigated in the marine mussels Mytilus californianus Conrad and M. edulis Linn. from the west coast of North America. In allopatry with M. californianus, M. edulis occurs throughout the intertidal zone; however, in microgeographic sympatry its ecological range is restricted to above the M. californianus mussel bed and to patches of substratum opened by natural disturbances within the bed. Over the same geographic scale, the broader-niched M. edulis shows greater among-locality genetic difference and greater levels of polymorphism than M. californianus at two enzyme loci. Genetic differentiation on a geographic basis was investigated in M. californianus at a single rock (on a scale of meters), on an island (on a scale of kilometers), throughout a strait (on a scale of 10² kilometers), and along the west coast of North America (on a scale of 10³ km). Differentiation was minimal over the west coast, and could be explained by microhabitat differences in a local area. The minimal differentiation in west coast mussels relative to strong geographic differentiation of M. edulis on the east coast of North America may be related to the steeper latitudinal thermal gradient on the east coast. Local ecologically related microgeographic variation can result in biased and misleading estimates of genetic heterogeneity but microgeographic variation at enzyme loci may not be due to selection at the loci investigated or may even be due to the breeding structure of the mussel populations, as suggested by Tracey et al. (1975).     

Occurrence of tri-n-butyltin-caused imposex in the North Pacific marine snail Nucella lima in Auke Bay,Alaska

Like female dog whelks (Nucella lapillus) in the Atlantic Ocean, females of the Pacific gastropod N. lima respond to low concentrations of tri-n-butyltin (TBT) by growing a penis and vas deferens. This condition, termed imposex, was found to occur in N. lima collected from August 1987 to May 1988 along a TBT pollution gradient associated with a marina in Auke Bay, Alaska. The suite of symptoms characteristic of imposex in N. lima was slightly different than in N. lapillus. Imposex, as measured by relative penis size (RPS) of females to males, increased from 0.0 to 34.27 along this gradient and as measured by vas deferens sequenceindex (VDS) increased similarly from 0.0 to 4.29. Concentrations of TBT in N. lima increased from below detection limits (about 0.010 g Sn g^-1 dry tissue wt) to 0.065 g Sn g^-1 dry tissue wt along the gradient. The gradient was determined by measuring TBT in whole-body tissues of bay mussels (Mytilus edulis). Concentrations of TBT in mussels increased from below detection limits to 0.833 g Sn g^-1 dry tissue wt in mussels from within the marina that was the major local source of TBT. Imposex, tissue TBT burden, and position along a TBT pollution gradient are significantly correlated (P<0.01) in N. lima. TBT was tested as a causative agent for imposex by exposing snails at a distant control site to TBT paint. After 1 mo exposure, 33% of the females grew a penis ranging in size from 0.2 to 0.8 mm. Our results generally corroborate those found for N. lapillus and indicate that imposex in N. lima is caused by environmental TBT exposure. We suggest that the RPS in the genus Nucella may be useful in monitoring TBT in coastal waters worldwide.     

Effects of copper on the latency of lysosomal hexosaminidase in the digestive cells of Mytilus edulis

Mytilus edulis collected from Tomales Bay, California, USA, during mid-winter 1979 were exposed to increased concentrations of dissolved copper under controlled laboratory conditions. A dose-dependent reduction in the latency of lysosomal hexosaminidase activity in digestive cells was induced after a 30 d exposure to copper. The half-time of the hexosaminidase staining reaction in sections of digestive gland from control mussels was 15.5 min; for mussels exposed to 25, 50, and 75 g Cu l^-1 it was 11.8, 8.5 and 5.5 min, respectively. In addition, the dyecoupled reaction product was seen earlier in sections from individuals exposed to 50 and 75 g Cu l^-1 (after 30 s) and 25 g Cu l^-1 (1 min) than in sections from control individuals (2.5 min). Copper accumulations were demonstrated histochemically to have the same distribution as the hexosaminidase reaction product, indicating that copper is sequestered in lysosomes. Copper concentrations in digestive gland tissue, were related to the concentrations of copper in the water to which the mussels were exposed.