Effect of nutrient supply on growth of blue mussels (Mytilus edulis) in a landlocked bay

The growth of blue mussels (Mytilus edulis) was studied in the landlocked bay Hopavågen in central Norway for 3 years, of which in 2 years (1998 and 1999) nutrients were added to increase the primary production. Nutrients (N:Si:P) were added daily from May to October in 1998 (molar ratio 15:5:4.1) and 1999 (molar ratio 16:8:1). The doses of nutrients correspond to 0.4 and 0.8 g P l^–1 day^–1 in 1998 and 1999 respectively. The growth of blue mussels (Mytilus edulis) in 1997 was followed at four depths (1, 2, 4 and 7 m). In 1998 and 1999 growth was followed at 2 and 10 m depths at four locations in Hopavågen and at a control station outside on the coast. The nutrient supply in 1998 only slightly increased the algal biomass (chlorophyll a), whereas mean daily primary production during the summer remained at the same level as the previous year. The increased nutrient supply in 1999 caused a nearly 50 and 100% increase in mean summer biomass and daily primary production, respectively. The growth of blue mussels in Hopavågen in 1997 and 1998 was within the same size range during the summer. In 1999 the shell length of blue mussels kept at 2 m depth was significantly higher than in the previous year at end of the growth season. The recorded growth was also significant higher than for mussels at 2 m depth at the control station. No difference in shell length was observed on mussels grown at 10 m depth in Hopavågen and in the control stations in 1998 and 1999. A higher tissue content was found in blue mussels grown at 2 m depth in Hopavågen, both in 1998 and 1999 when compared to the control groups. At 10 m depth no differences were recorded.     

A new approach for the assessment of stochastic variation: analysis of behavioural response in blue mussel (Mytilus edulis L.)

One of the most effective techniques for evaluating stress is the analysis of developmental stability, measured by stochastic variation based particularly on fluctuating asymmetry, i.e. a variance in random deviations from perfect bilateral symmetry. However, the application of morphological methods is only possible when an organism lives under testing conditions during a significant part of its ontogenesis. Contrary to morphological characters, behavior can change very fast. Consequently, methods based on behavioural characters may have advantages over more traditional approaches. In this study we describe the technique of assessing stochastic variation, using not morphological, but behavioural characters. To measure stochastic variation of behavioural response, we assessed the stability of the isolation reaction of blue musselMytilus edulis at regular changes of salinity. With increasing temperature from +12°C to +20°C stochastic variation of the isolation reaction increased, which is a common response to change of environmental conditions. In this way, we have developed a method of assessing stochastic variation of behavioural response in molluscs. This method may find a great range of applications, because its usage does not require keeping animals in tested conditions for a long time.     

Documentation of sites of intertidal blue mussel (Mytilus edulis L.) beds of the Lower Saxonian Wadden Sea, southern North Sea (as of 2003) and the role of their structure for spatfall settlement

Field surveys (dating back to 1950) and aerial photograph series (dating back to 1966) were evaluated to determine sites of intertidal blue mussel (Mytilus edulis) beds at the Wadden Sea coast of Lower Saxony. Maps were prepared indicating sites of blue mussel beds during the last decades. A table gives additional information on the presence (or absence) of blue mussel beds at each site at the time of large-scale surveys. Altogether 187 sites of M. edulis beds were recorded in the investigation area. In spring 1996, there were still only 19 sites where mussel beds still occurred, although at 51 sites residual mussel-bed structures were present, e.g. shell bases of former beds or protruding patches (which had been occupied by M. edulis before the beds vanished) and open spaces. At that time, the majority of the sites contained neither mussel beds nor mussel-bed structures. The analysis of recent data confirmed that mussel larvae have preferred to settle in sites of present mussel beds and sites with bases of former mussel beds. There was no preferential selection of one of these categories (settled beds vs. shell bases). On the other hand, the presence of mussel beds or mussel bed structures is not obligatory for settlement, because sites without those structures were also re-settled by the spatfall in 1996, even though on a smaller scale.     

The application of combined tissue residue chemistry and physiological measurements of mussels (Mytilus edulis) for the assessment of environmental pollution

The rationale for the use of combined tissue residue chemistry and physiological energetics measurements of Mytilus edulis in the assessment and monitoring of environmental pollution is outlined. Laboratory derived relationships between the concentration of toxicants in tissues and sublethal responses (eg. feeding, respiration and growth rate) provide a toxicological database for the interpretation of physiological responses measured in the field. The role of quantitative structure-activity relationships (QSAR's) in establishing tissue concentration-effect relationships for organic contaminants is discussed. The application of this approach is illustrated with reference to two field studies, a monitoring programme in the Shetlands and a practical biological effects workshop in Oslo.     

Protection against suspended sand: the function of the branchial membrane in the blue mussel Mytilus edulis

Blue mussels (Mytilus edulis) living in estuaries have to cope with varying concentrations of suspended sand. Sand flowing through the inhalant siphons comes into the infrabranchial chamber. The inhalant siphon can be partially closed by the branchial membrane. As a result the inward flow decreases, and suspended sand sinks and can be eliminated. Experiments with mussels from three ecologically different locations showed about the same response of the branchial membrane on contact with suspended sand. The presence and function of the branchial membrane appears to be an adaptation of mussels to their estuarine environment.     

Short-term impact of blue mussel dredging (Mytilus edulis L.) on a benthic community

The short-term effect of mussel dredging in a brackish Danish sound was studied. A commercial dredging track was identified and an analysis of the species composition inside the track and at an adjacent control area showed that dredging changed the community structure by reducing the density of polychaetes. In order to investigate the extent and the duration of the dredging impact experimental dredging was conducted. The experimental dredging removed 50% of the mussels in two dredged areas. Immediately after dredging, a significantly lower number of species was measured inside the mussel beds in dredged areas compared to control and boundary areas. This effect lasted for at least 40 days. The analysis of the species composition showed that the dredged area had a significantly lower density, particularly of polychaetes compared to the boundary area. An increased number of species was recorded outside the mussel beds just after dredging, but this effect lasted for less than 7 days. After dredging, brown shrimps, C. crangon invaded the dredged areas. This species is an important predator of smaller invertebrates, and it is suspected that it was feeding on small vulnerable polychaetes exposed at the sediment surface after dredging. The dredging process was observed to form 2–5-cm deep furrows in the seabed, but the sediment texture and the organic content of the sediment was not affected. The biomass accumulation of individual blue mussels was significantly lower in the dredged area compared to the boundary area. This indicates that the disturbance of the mussel bed structure reduced growth and that the lowering of intraspecific food competition caused by a reduced density of mussels did not increase the accumulation of biomass in the mussels which remained in the dredged area.     

The role of the blue mussel Mytilus edulis in the cycling of nutrients in the Oosterschelde estuary (The Netherlands)

The fluxes of particulate and dissolved material between bivalve beds and the water column in the Oosterschelde estuary have been measured in situ with a Benthic Ecosystem Tunnel. On mussel beds uptake of POC, PON and POP was observed. POC and PON fluxes showed a significant positive correlation, and the average C:N ratio of the fluxes was 9.4. There was a high release of phosphate, nitrate, ammonium and silicate from the mussel bed into the water column. The effluxes of dissolved inorganic nitrogen and phosphate showed a significant correlation, with an average N:P ratio of 16.5. A comparison of the in situ measurements with individual nutrient excretion rates showed that excretion by the mussels contributed 31–85% to the total phosphate flux from the mussel bed. Ammonium excretion by the mussels accounted for 17–94% of the ammonium flux from the mussel bed. The mussels did not excrete silicate or nitrate. Mineralization of biodeposition on the mussel bed was probably the main source of the regenerated nutrients.From the in situ observations net budgets of N, P and Si for the mussel bed were calculated. A comparison between the uptake of particulate organic N and the release of dissolved inorganic N (ammonium + nitrate) showed that little N is retained by the mussel bed, and suggested that denitrification is a minor process in the mussel bed sediment. On average, only 2/3 of the particulate organic P, taken up by the mussel bed, was recycled as phosphate. A net Si uptake was observed during phytoplankton blooms, and a net release dominated during autumn. It is concluded that mussel beds increase the mineralization rate of phytoplankton and affect nutrient ratios in the water column. A comparison of N regeneration by mussels in the central part of the Oosterschelde estuary with model estimates of total N remineralization showed that mussels play a major role in the recycling of nitrogen.     

Turnover rate of metallothionein and cadmium in Mytilus edulis

The results demonstrate the first attempt to determine metallothionein turnover in the whole soft tissues of mussels Mytilus edulis exposed to cadmium. Half-lives for metallothionein and cadmium are 25 and 300 days, respectively. As metallothionein degrades the released cadmium induces further synthesis of the protein, to which the metal becomes resequestered. The slow metallothionein turnover rates (compared with mammals) and the lack of significant cadmium excretion testify to the relatively stable nature of the cadmium-metallothionein complex in these invertebrates and supports the view of a detoxifying role for metallothionein in the mussels.     

A methodology for screening haemolymph of intertidal mussels, Mytilus edulis, using FT-IR spectroscopy as a tool for environmental assesment

Developing effective, rapid and inexpensive methods for monitoring and conserving aquatic resources is an important issue for environmental managers. This study focuses on Mytilus edulis, a keystone species of many coastal marine communities, which is frequently used as a biomonitor for a range of pollutants. Recent advances in post-genomic technologies have provided new methods of biochemical screening, and Fourier transform-infrared spectroscopy (FT-IR) is one such method that could enable bioindicator species to be used for environmental assessment. This paper develops a methodology to apply the FT-IR approach to marine intertidal M. edulis and addresses three methodological issues: First, the optimum physical location for biofluid sampling is examined (i.e. laboratory versus field). Secondly, the effects of transportation of frozen biofluid sampling from either the field-site or laboratory to the analytical facility are considered. Finally, the effect of repeated FT-IR measurements on collected M. edulis haemolymph samples is examined. From these results we suggest sampling haemolymph from M. edulis at the top of the shore prior to immediate snap-freezing in liquid nitrogen. Sample transportation can occur on ice for up to eight hours before storage at −80 °C. FT-IR measurements should occur within three months of collection and samples should not be used or thawed more than twice. We show how this method can be used to differentiate successfully between four different estuarine environments. Ultimately, through addressing these methodological questions, we provide a protocol to allow efficient sampling and FT-IR measurement of M. edulis as collected from the intertidal areas of rocky and muddy shores. We conclude that due to current monitoring needs presented by the European Water Framework Directive such an approach could prove to be an invaluable future tool for assessing coastal water quality.     

On the ultrastructure of polypeptide hormone-producing cells in the gut of the ascidian,Ciona intestinalis L. and the Bivalve,Mytilus edulis L.

Summary Ultrastructural evidence has been found for the presence of polypeptide hormone-producing cells in the gut of Ciona intestinalis L. and Mytilus edulis L. which do not appear to have been described before. Due to their localization and ultrastructural characteristics, it is suggested that the cells in Mytilus edulis probably produce an insulin-like substance and that some of these cells in Ciona intestinalis may produce 5-HT (5-Hydroxytryptamine). In each species only one granulated cell type can be observed. The granules, which are electron dense and membrane bound, also show a halo. The average diameter of the granules is 100–200 nm for Ciona and 200–400 nm for Mytilus.I thank Mr. G. Bargsten, M.A., Dept. of Marine Zoology, University of Kiel, for the supply of the animals     

Differential binding of lectins to haemocytes of the mussel Mytilus edulis

Summary Pre- and post-embedding techniques were used to investigate the ultrastructural binding of a range of lectins to the haemocytes of the mussel Mytilus edulis. Direct and indirect labelling procedures were employed using colloidal gold and ferritin-labelled lectins, or biotinylated lectins followed by gold-labelled streptavidin. Cell surface receptors were present for lectins from Helix pomatia (HPA), Helix aspersa (HAA), Triticum vulgaris (WGA) and Tetragonolobus purpureas (TPA). Double labelling of haemocytes with HPA and WGA demonstrated binding sites for both lectins on the plasma membrane of the majority of haemocytes. Endocytosis of colloidal gold-labelled HPA was observed for unfixed haemocytes. Three classes of haemocyte were identified by use of morphological criteria: hyalinocytes; granulocytes containing small granules; and granulocytes containing large granules. Lectin binding showed the small granules of the granulocytes to be HPA-positive and the large granules of the granulocytes to be WGA-positive. The WGA-positive granules demonstrated a differential pattern of binding according to granule size. Binding sites for the lectin from Arachis hypogaea (PNA) were not demonstrated on the cell surface, but did show an affinity for the heterochromatin region of the nucleus in post-embedding protocols.     

Incidence of hemocytes and parasites in coastal populations of blue mussels (Mytilus edulis)--testing correlations with area,season, and distance to industrial plants

Blue mussel hemocytes (cells with immunoresponse activities) are suggested as indicators of anthropogenic contamination. We compared hemocyte numbers, granulocytoma (aggregated hemocytes), and parasites among populations of mussels from different areas of Skagerrak (a north and a south), seasons (summer and autumn), and impact levels (close or far from industrial activities). Seasonal hemocyte numbers were larger in the north compared to the south. Northern unimpacted populations had higher hemocyte numbers than populations close to industries, while no differences were found in the south. More uneven tissue distributions were found in populations far from industries in the north area and in populations close to industries in the south area. Parasites were more common in northern mussels than in southern, but no relationship to impact level was found. Mussels with granulocytoma, however, were found in all populations from the impacted sites while in none of the other populations suggesting granulocytoma as a possible indicator of industrial impact.     

Innervation of the anterior byssal retractor muscle in Mytilus edulis L.

Summary Studies on the intrinsic innervation of the anterior byssal retractor muscle (ABRM) in Mytilus edulis L. were continued at the ultrastructural level. Electron micrographs show nerve processes ensheathed by glio-interstitial cells running between muscle fibers. The glio-interstitial cells may represent all the types of osmiophilic cells previously described by the light microscopic ZIO technique in the anterior byssal retractor muscle.     

Age-dependence of metabolism in mussels Mytilus edulis (L.) from the White Sea

Age structure, natural mortality and growth, as well as age- and size-dependent changes in parameters of energy metabolism were studied in blue mussels Mytilus edulis (L.) from the White Sea. Mussels were sampled in August (Summer sample, SS) and October (Autumn sample, AS) and contained animals of three size groups, 2-9 years old. Field data showed an increase of mortality of mussels and strong decrease in growth rates after 6 years of age. Absolute tissue growth increment (AI) reconstructed from winter growth marks on the shells decreased with age and was strongly size-dependent, while relative tissue growth increment (RI) did not depend on size of the animals. Respiration rates and citrate synthase activity demonstrated power regression versus tissue weight with regression coefficients -0.231 and -0.170, respectively. After weight correction both parameters showed a decrease with increasing age. ATP and phosphagen levels also showed a pronounced decrease in animals older than 5-6 years despite considerable differences in the absolute values of both parameters in SS and AS. pH(i) in mussels was also age-dependent and decreased with increasing age after 5 years. In air exposed mussels, pH(i) was reduced only at young age such that pH(i) was low and constant within the whole age range. Our data give evidence that aerobic metabolic rate in M. edulis from the studied population declines when animals reach an age of about 6 years. The decrease in oxygen consumption reflects the drop in mitochondrial respiration, which is mirrored by the decrease in CS activity. A concomitant fall in ATP turnover may include a downregulation of the mechanisms of acid-base regulation. pH(i) will then approach equilibrium indicated by lower pH(i) values in older animals. Our data suggest that intrapopulational comparisons of physiological parameters in mussels should take into account age composition of compared samples.     

Lipopolysaccharide and opioids activate distinct populations of Mytilus edulis immunocytes

Summary Studies in Mytilus edulis have indicated that immunoregulatory activities comoparable to those in vertebrates also exist in invertebrates. Mytilus immunocytes resemble cells of the vertebrate monocyte/macrophage lineage and are activated by similar substances. We searched for differential effects of opioids on these cells in comparison with those of lipopolysaccharide (LPS), in order to determine if different subpopulations of immunoactive hemocytes are involved. We showed that Mytilus immunocytes respond to LPS in a fashion similar to that in vertebrate granulocytes by flattening, and increasing in cellular perimeter and mobility, that LPS administered in vivo results in a lowering of the number of free hemocytes that can be obtained from the animal, and that distinct immunoactive cell populations seem to exist since apparently different subsets of cells react when exposed to LPS or opioids and the opioid antagonist naloxone.     

Impact of suspended mussels (Mytilus edulis L.) on plankton communities in a Magdalen Islands lagoon (Québec, Canada): A mesocosm approach

The Grande-Entrée Lagoon (Magdalen Islands, Canada) has supported mussel (Mytilus edulis) cultivation for the last 25 years. Algal biomass in this lagoon is relatively low while heterotrophic plankton biomass is high. Although often considered herbivorous, it is known that filter-feeding bivalves can consume various types of food, from bacteria to zooplankton. We hypothesize that along with phytoplankton, heterotrophs constitute an important food resource for the Grande-Entrée mussels. In situ mesocosm experiments were undertaken at different seasons using short socks filled with mussels from the same cohort taken from an aquaculture farm, in order to determine the impact of cultured mussels on local plankton communities and assess the role of heterotrophs. Filtration activity by the mussels and associated epibionts present in the socks was expressed as clearance rates (CR). The average CR over all taxa was lowest in June and highest in October. Diatoms, dinoflagellates and heterotrophic protists constituted the bulk of planktonic carbon removed by mussels. While smaller-sized taxa contributed little (< 5%) to mussel carbon intake, large-sized heterotrophs (namely ciliates) contributed 69 to 88%. Taxon-marker pigment analyses generally confirmed these observations for groups containing phototrophic pigments. The high heterotrophic biomass retained by mussels indicates they are a major food source for mussels in this environment and should be considered both in the evaluation of mussel feeding and in assessing the influence of cultured mussels on local plankton ecosystems.     

Three-dimensional crystal structure and enzymic characterization of beta-mannanase Man5A from blue mussel Mytilus edulis

Endo-beta-1,4-d-mannanase is the key depolymerizing enzyme for beta-1,4-mannan polymers present in the cell walls of plants and some algae, as well as in some types of plant seeds. Endo-1,4-beta-mannanase from blue mussel Mytilus edulis (MeMan5A) belongs to the glycoside hydrolase (GH) family 5 enzymes. The MeMan5A structure has been determined to 1.6A resolution using the multiple-wavelength anomalous dispersion method at the selenium K edge with selenomethionyl MeMan5A expressed in the yeast Pichia pastoris. As expected for GH 5 enzymes, the structure showed a (betaalpha)(8)-barrel fold. An unusually large number of histidine side-chains are exposed on the surface, which may relate to its location within the crystalline style of the digestive tract of the mussel. Kinetic analysis of MeMan5A revealed that the enzyme requires at least six subsites for efficient hydrolysis. Mannotetraose (M4) and mannopentaose (M5) were shown to interact with subsites -3 to +1, and -3 to +2, respectively. A clear kinetic threshold was observed when going from M4 to M5, indicating that the +2 subsite provides important interaction in the hydrolysis of short oligomeric mannose substrates. The catalytic centre motif at subsite -1 found in superfamily GH clan A is, as expected, conserved in MeMan5A, but the architecture of the catalytic cleft differs significantly from other GH 5 enzyme structures. We therefore suggest that MeMan5A represents a new subfamily in GH 5.