Hybridisations between Mytilus edulis and Mytilus galloprovincialis and performance of pure species and hybrid veliger larvae at different temperatures

The mussels Mytilus edulis L. and Mytilus galloprovincialis Lamark hybridise naturally in the wild along the Atlantic coast of Europe producing a patchwork of mixed pure species and hybrid populations. Individuals of both species were spawned in the laboratory and were hybridised in a series of reciprocal crosses. After 72 h, the proportion of eggs which developed into larvae (%yield) and the proportion of those larvae which had a normal veliger morphology (%normality) were estimated and compared between pure species and hybrid families. There were no significant differences in %yield or %normality between pure species and hybrids, but significant differences were evident between the offspring from different parents irrespective of whether they were hybrids or pure species. Therefore confirmation of hybrid heterosis in laboratory studies should not be based on a single, or a few reciprocal crosses. Hybrid and pure species veliger larvae were grown for approximately 4 weeks at 10, 14 or 20 °C. In all trials, pure M. galloprovincialis larvae grew significantly faster at 20 °C than either reciprocal hybrid or pure M. edulis larvae. Irrespective of temperature, in general, hybrid larvae grew slower than larvae of either pure species. Increased exposure to planktonic predation due to slow growth can be interpreted as selection against hybrids and this may play a role in the structure and distribution of mixed pure species and hybrid populations.     

Soluble and surface-bound aminopeptidase in eosinophilic blood cells from Mytilus edulis

This study focused on soluble and surface-bound aminopeptidase (AP) in hemocytes from Mytilus edulis and on the identification of the enzyme-producing blood cells. The cell extract hydrolyzed alanine p-nitroanilide (Ala-pNA) with an optimum between pH 6.4 and 7.0. Following native gradient PAGE of extract, alanyl methoxy-naphthylamide (AMNA) was converted by one band with an estimated molecular weight of 375 kDa; it included at least ten putative AP-isozymes with isoelectric points between pH 4.5 and 5.8. In addition to this soluble form, electron microscopy revealed simultaneous conversion of AMNA on the surface of blood cells. Individual mussels expressed AP-molecules in 23-39% of their hemocytes. These cells were shown to represent eosinophilic granulocytes.     

Growth in length of Mytilus edulis L. fed on different algal diets

Mytilus edulis L. was fed on different algal diets and the increase in shell length was measured every 12–24 h. The mussels respond within 12 h to major changes in the diet. When pre-starved mussels were fed every 24 h with monocultures of Tetraselmis suecica (Kylin) Butch, there was a pronounced lag period followed by a linear increase in shell growth rate. When both pre-starved and pre-fed mussels were fed on equal rations of T. suecica with concentrations ranging from 2.5 to 10.0 × 10⁷ cells · 1^?1, the growth rate levelled off at about the same rate. Within the same range of concentrations there was a linear correlation between final growth rates and algal cell concentration. Feeding with monocultures of Isochrysis galbana (Parke) or Thalassiosira pseudonana Hasle et Heimdal (Hustedt) gave approximately the same shell growth as with Tetraselmis suecica alone, while combinations of these three algal species produced significant synergistic effects. When filtrate only from T. suecica cultures was supplied to the mussels there was a rapid initial stimulation of the shell growth. Centrifugated cells of T. suecica which were resuspended in filtered sea water, homogenized and sonicated to rupture the cells, gave 13% less growth than with untreated cells (P < 0.05). The use of the accurate laser diffraction method for length growth measurements may greatly reduce the time and effort involved in growth experiments.     

Characterization of metal-containing granules from the kidney of the common mussel, Mytilus edulis

The metal-containing membrane-limited granules which make up some 20% of the cellular volume of the kidney cells of the marine mussel, Mytilus, have been purified and partially characterized by chemical and histochemical analysis. They contain an inorganic component (10% of their dry weight), principally iron, zinc and calcium, associated with sulphur phosphorus together with a pigmented organic component which has the histochemical and spectroscopic of lipofuschin. Microscopic and enzymic evidence indicates that the lipofuschin is present as a result of lysosomal degradation and peroxidation of cellular membranes. The granules may thus be regarded as tertiary lysosomes or residual bodies. The ash content of these lysosomes is 10-fold higher than those from mammalian sources. For this reason, Mytilus may be a useful model system for the study of intralysosomal metal accumulation.     

The ecology of Mytilus edulis L. in exposed rocky intertidal communities

Information is given to show that Mytilus edulis L. is an important component of exposed rocky coast communities of the Western Hemisphere. It is abundant on the exposed coast of Washington State with a potential distribution extending from + 10.6 ft ( + 3.2 m) down to at least 10 m depth: it is not continuous along this entire gradient. It dominates a band from +10.6 ft down to +9.6 ft ( + 3.2 to + 2.9 m) tidal height where it usually intergrades with its congener M. californianus Conrad. Its upper limit appears to be determined by desiccation stress and the lower probably by competition and or predation.In the mid-intertidal zone, from + 5.6 to + 9.6 ft ( + 1.7 to 2.9 m), where M. californianus is dominant, M. edulis occurs in patches of cleared space which usually form in the M. californianus bed matrix in winter months. Here, it acts as an opportunistic species by colonizing quickly and growing rapidly to reproductive size. It is preyed upon by Thais spp. which prefer it to Mytilus californianus and eventually is eliminated from these patches. It also occurs, sometimes abundantly, in tufts of the red alga Endocladia muricata (Postels & Ruprect) J. Agardh at high and mid-intertidal heights.In low intertidal and subtidal regions, to ≈ 10 m depth, Mytulis edulis can be found in refuges of filamentous or intricately arranged substrata which generally offer protection from predation. These are represented by a wide variety of forms such as hydroids. bryozoans, filamentous algae, coralline algae, and kelp with convoluted holdfasts or complex stipes.     

Larval recruitment of the blue mussel Mytilus edulis: The effect of flow and algae

The mussel Mytilus edulis settlement and distribution was studied on plastic panels with manipulated flow regime (faired, bluff, split and angled) with or without water soluble metabolites of the green alga Cladophora rupestris. The panels were exposed vertically on a device (hydrovane) that ensures their constant orientation in the current during the peak of larval settlement at 1 m depth. In order to investigate larval distribution on the panels, half of them were coated with a silicone vacuum grease that prevents larvae from de-attachment. This grease was not toxic and did not attract or repel larvae. Low densities of larvae on the un-greased plates compared to the greased ones suggested that some of larvae left the substratum. The blue mussel larvae initially settled in regions of reduced shear velocity and then redistribute to the regions of high shear velocity. The presence of the alga increased the density of blue mussel larvae and changed their distribution on the panels. Overall, our results demonstrated that larval recruitment of M. edulis is an active process affected both by boundary-layer hydrodynamics and algal waterborne compounds.     

Exposure of the blue mussel, Mytilus edulis, to gold nanoparticles and the pro-oxidant menadione

Relatively little is known about how gold nanoparticles (GNP) might interact in vivo with marine organisms. Mytilus edulis was exposed (24 h) to ~ 15 nm GNP, menadione and both compounds simultaneously (GNP/menadione). GNP was detected by inductively coupled plasma-optical emission spectroscopy mainly in digestive gland of samples exposed to GNP though not GNP/menadione, perhaps due to impaired feeding. Thioredoxin reductase activity and malondialdehyde levels were determined in all tissues. Thioredoxin reductase inhibition was detected only in digestive gland exposed to menadione whilst malondialdehyde levels did not vary in response to treatment in all tissues. GNP caused a decrease in the reduced/oxidized glutathione ratio in digestive gland, but no difference was found in other tissues or for other treatments. One dimensional electrophoresis of proteins containing thiol groups was performed in all tissues and revealed a reduction in protein thiols for all treatments in digestive gland. Two dimensional electrophoresis of digestive gland extracts, from GNP and control groups, showed decreased levels of thiol proteins in response to GNP which we attribute to oxidation. Our results suggest that GNP causes a modest level of oxidative stress sufficient to oxidize thiols in glutathione and proteins but without causing lipid peroxidation or induction of thioredoxin reductase activity.     

Geographical variation in leucine aminopeptidase in Mytilus edulis L. From the Irish coasts

Phenotypic variation in leucine aminopeptidase has been demonstrated in Mytilus edulis L. from twelve localities in Ireland. It is proposed that one locus with three common and two rare alleles is responsible for one group of variants. The frequencies of the common alleles are significantly correlated with the relative amount of wave action (exposure) at the sites investigated. M. edulis seems to exhibit an above average amount of genetic variation thereby paralleling the variations in growth rate, shell morphology, filtration rate, and certain kinetic parameters previously recorded amongst individuals of this species. In view of the apparently close correlation between genotype and environmental conditions and since genotype in other species is known to control survival and growth rate, genetic studies of the kind described here could clearly have important implications for the commercial mussel fishery.     

The passage of two species of micro-algae through the gut of Mytilus edulis L.

The contents of fecal ribbons from Mytilus edulis L. fed either the unicellular flagellate Tetraselmis suecica (Kylin) Butch., or the diatom Thalassiosira pseudonana (Hasle and Heimdal) were examined for their particle size spectra. Contents of fecal ribbons were dispersed by shaking in filtered sea water and analyzed by a Coulter Counter. The flagellate can pass through the mussel's gut without any significant volume changes, implying that its digestion is largely intracellular (after phagocytosis). In contrast, the diatom does undergo volume changes, probably a result of disintegration of cells and aggregation of cells and cell fragments. Undigested flagellate cells can become resuspended in significant quantities after their defecation by the mussel.     

Prey selection within a size-class of mussels, Mytilus edulis, by oystercatchers, Haematopus ostralegus

By comparing the shells of those mussels Mytilus edulis that had been opened by oystercatchers Haematopus ostralegus with mussels of similar size that had not been opened, it was shown that oystercatchers that break into their prey by hammering a hole in the shell selected between prey within a size-class. Ventral hammerers selected mussels that were relatively thin on the ventral surface, were brown in colour and carried few barnacles. Dorsal hammerers selected eroded mussels with thin dorsal shells. Stabbing oystercatchers did not select for thin-shelled prey. In conjunction with the great individual variation in flesh content between mussels of the same length, prey size in this case can be only a poor predictor of prey profitability.     

Light microscopical studies of the acute toxic effects of N-nitrosodimethylamine on the marine mussel, Mytilus edulis

Mussels, Mytilus edulis, exposed to N-nitrosodimethylamine (DMN), 100 mg DMN/liter sea water, exhibited the following tissue responses: Congestion of the larger vessels in the Leydig cell tissue and infiltration of the tissue with numerous hemocytes, well-demarcated lesions composed of hemocytes in the Leydig cell tissue, degeneration of the epithelial lining of the ducts of the digestive diverticula, and congestion of the branchial vessels with numerous hemocytes accompanied by distortion of the cilia and the ciliated columnar epithelium of the gills. Furthermore, deposition of collagen-like material in the walls of the vessels was observed. DMN also induced spawning prior to the normal spawning time.     

The effect of salinity change on the heart rate of Mytilus edulis specimens from different ecological zones

The heart rate of specimens of Mytilus edulis (L.) both from the sublittoral and littoral zones exposed to normal and altered salinity was investigated in a long-term experiment. The heart rate was monitored by a non-invasive method for nine days. The heart rate of sublittoral mussels was higher than that of littoral ones. This suggests a higher level of metabolic activity in sublittoral mussels. When exposed to moderate hyposalinity (15 g l⁻¹) M. edulis from both zones showed a significant decrease in the heart rate with respect to the control salinity (25 g l⁻¹), but sublittoral mussels had a more prominent bradycardia. The heart beat quickly accelerated in all organisms when they were returned to the control salinity medium. Throughout the experiment, heart contractions halted with distinct periodicity in all mussels.     

Influence of abiotic factors on bacterial proliferation and anoxic survival of the sea mussel Mytilus edulis L.

The effect of several abiotic factors (salinity, temperature and pH) on bacterial proliferation and survival time of the sea mussel Mytilus edulis L. were studied under anoxic incubations. In addition, the presence in the incubation media of ammonium and the volatile fatty acids propionate and acetate, both excreted fermentation products of the bivalve, was tested.Anoxic incubations with seawater diluted with demineralised water showed at the lowest salinity (50% seawater, SW) a significant increase in the capacity of M. edulis to survive anoxia as compared to both 75% SW and control [100% SW, corresponding to 32 practical salinity units (psu)]. Formation of biotic sulphide and ammonium occurred in all incubations. However, bacterial proliferation was postponed by 2-3 days at lowest salinity and accordingly, concentrations of both compounds were lower. Anoxic survival profiles of mussels collected from different habitats in the Dutch Scheldt area, characterised by differences in salinity (range from 17 to 31 psu), corresponded with the above salinity effect. Walsoorden mussels (17 psu) showed the longest (P<0.001) survival time under anoxia (LT₅₀=17.2 days) as compared with Paulina (27 psu) and Wemeldinge (31 psu) mussels (LT₅₀=12.8 and 9.8 days, respectively). Condition index (ratio of soft body weight to shell volume) was not correlated with anoxic survival time in untreated mussels, although this was clearly the case when the antibiotic chloramphenicol was added to the anoxic seawater.Acidification of the anoxic incubation medium had a positive effect on survival time. LT₅₀ values significantly (P<0.001) increased from 10.2 days at pH 8.1 to 11.6 and 11.5 days at pH 7.3 and 6.5, respectively. Biotic sulphide and ammonium accumulation as well as bacterial numbers were significantly lower at pH 7.3 and 6.5 as compared with pH 8.1. Anoxic incubations at 10 °C (LT₅₀=12.0 days) strongly increased survival time as compared to 18 °C (LT₅₀=5.9 days). The benefit of antibiotic addition was also stronger at lower temperature (10 °C).Addition of both propionate and acetate (0.5 mM) displayed no effect on mortality of mussels under anoxia, but ammonium (0.5 mM) caused a negative effect (P<0.001). Biotic sulphide and ammonium concentrations measured in both volatile fatty acid incubations were lower than the control situation, as well as total bacterial numbers.This study shows that environmental factors play a significant role in determining the course of bacterial infection and death of bivalves exposed to anoxia.     

Effects of salinity on the respiration and heart rate of the common mussel, Mytilus edulis L., and the black chiton, Katherina tunicata (Wood)

The rate of oxygen consumption of stepwise acclimated Mytilus edulis L. increased linearly from 30 to 10‰ salinity (S) while that of Katherina tunicata (Wood) was not significantly different between 10 and 30‰ S. Heart rate was 21–22 and 17–18 beats m^?1 in Mytilus edulis and Katherina tunicata, respectively, and no difference was found in the heart rate of either species acclimated stepwise to 10, 20 or 30‰ S. The average oxygen consumption rate of Mytilus edulis exposed to 12 h, 30-10-30 and 10-30-10‰ S cycles of fluctuating salinity was significantly lower than the respective control rate: there was a similar response during the 30-10-30‰ S cycle in Katherina tunicata. The respiration rate of Mytilus edulis and Katherina tunicata declined as salinity deviated from the control salinity and increased as salinity returned to the control salinity. The rate of oxygen consumption by K. tunicata varied directly with the ambient salinity during the 10-30-10‰ S cycle. The average heart rate of Mytilus edulis was significantly lower during cyclic changes in salinity than at the respective control salinities; a similar relationship existed for Katherina tunicata during the 10-30-10‰ S cycle. Heart rate of Mytilus edulis varied in a parallel manner with oxygen consumption during both cycles. Katherina tunicata heart rate was relatively constant and could not be fitted to a regression line during the 10-30-10‰ S cycle. The normalized heart rate increased to 113% of control at 10‰ S of the 30-10-30‰ S cycle and returned to the control rate by 12 h. The oxygen consumption and heart rate of these two species are not directly coupled to regulation of water volume because different responses are observed with respect to salinity although there is poor water volume regulation in both species.     

Experimental test of biodeposition and ammonium excretion from blue mussels (Mytilus edulis) on eelgrass (Zostera marina) performance

Blue mussels and eelgrass have been found to coexist in many locations. However, knowledge of the interactions between these species is limited. Two experiments were conducted in the laboratory, a “Deposit” and an “Epiphyte” experiment. The Deposit experiment examined possible effects of increasing load of blue mussel (Mytilus edulis) biodeposits on sediment biogeochemistry and eelgrass (Zostera marina) performance. Z. marina mesocosms received normal or high loads of mussel biodeposits (Normal and High), while no biodeposits were added to the Control. High dosage had overall negative effects on Z. marina, which was reflected as lower leaf numbers and biomass and accumulation of elemental sulphur in rhizomes. The sediment biogeochemical conditions were altered, as the mussel biodeposits enhanced sulphate reduction rates and increased sulphide concentrations in the porewater, which may result in sulphide invasion and reduced growth of Z. marina.In the Epiphyte experiment effects of mussel excretion, with particular emphasis on ammonium, on the growth of Z. marina and their epiphytes were examined. A thick cover of epiphytes developed on Z. marina growing together with M. edulis, and the relative growth rate was reduced with 20% compared to plants from control without mussels. Overall the experiments showed negative effects on Z. marina growing together with M. edulis, thereby supporting a preceding field study by Vinther et al. [Vinther, H.F., Laursen, J.S., Holmer, M. 2008. Negative effects of blue mussel (Mytilus edulis) presence in eelgrass (Zostera marina) beds in Flensborg fjord, Denmark. Est. Coast Shelf. Sci. 77, 91-103.].     

Bacicyclin,a new antibacterial cyclic hexapeptide from Bacillus sp. strain BC028 isolated from Mytilus edulis

A new cyclic hexapeptide, cyclo-(Gly-Leu-Val-IIe-Ala-Phe), named bacicyclin (1), was isolated from a marine Bacillus sp. strain associated with Mytilus edulis. The sequences of the amino acid building blocks of the cyclic peptide and its structure were determined by 1D- and 2D-NMR techniques. Marfey’s analysis showed that the amino acid building blocks had L-configuration in all cases except for alanine and phenylalanine, which had D-configuration. Bacicyclin (1) exhibited antibacterial activity against the clinically relevant strains Enterococcus faecalis and Staphylococcus aureus with minimal inhibitory concentration values of 8 and 12?µM, respectively. These results demonstrate the potential of marine bacteria as a promising source for the discovery of new antibiotics.     

The contribution of marine snow to the particle food supply of the benthic suspension feeder, Mytilus edulis

In order to examine the importance of the settling of large particles to the food supply and feeding behavior of a benthic culture of the blue mussel, Mytilus edulis, we investigated the tidal dynamics of large (>0.5 mm diameter) marine aggregates, commonly known as marine snow, during three tidal cycles in July 1998 at a shallow, subtidal, low current flow regime site along the coast of Maine (Shorey Cove, Roque Island, Englishman's Bay, Maine).In situ, optically measured marine snow showed a distinct tidal signal displaying an increase in size and abundance through high tide with a peak on the early ebb tide as it settled to the bottom. Marine snow volume ranged an order of magnitude through the tidal cycle, from under 8 to over 80 mm³ l⁻¹. An increase in the in situ marine snow volume corresponded with an increase in benthic mussel feeding activity (from 20% to 60% of maximum exhalant siphon area, which is an estimate of pumping rate) and maximum rates of pseudofeces production by the mussels during periods of low tidal current speeds. In contrast, mussels from the same population feeding on surface waters in shipboard chambers produced no pseudofeces and had high pumping rates (80-100% maximum exhalant siphon area) over the whole tidal period. A second peak in benthic mussel pumping rates also occurred during flood tide.Food quality was lower in the bottom waters due to significantly higher particulate inorganic matter (PIM, >2 mg l⁻¹) when compared with the surface waters. PIM accounted for 95% of the total settled mass flux of 3.4 g m⁻² day⁻¹ measured in sediment traps deployed 1 m off the bottom, with organic carbon representing only 2.5% of the mass flux during the mid-summer conditions. At low-current sites such as Shorey Cove, Roque Island, Maine, the settling of marine snow provides an important additional source of food, albeit of low quality, to benthic populations of blue mussels.     

The mechanism of calcium transport in mitochondria isolated from the marine mussel, Mytilus edulis (L.)

Isolated mussel mitochondria produced a less pronounced transient stimulation of respiration upon the addition of Ca²⁺ in a reaction medium containing Pi and a slower rate of Ca²⁺ transport compared to rat liver mitochondria. The initial rates of Ca²⁺ transport in the absence of Pi were more similar and both types of mitochondria possessed a sigmoidal relationship between the initial rate of Ca²⁺ transport and the free Ca²⁺ concentration $\text{(‘Km’ ? 5μM)}$. Ruthenium red produced an equal maximal inhibition of the initial rate of Ca²⁺ transport in both types of mitochondria but mussel mitochondria were rather more resistant to the inhibitor. The major difference found was that approximately 15 nmoles La³⁺ mg protein^?1 was required to produce maximal inhibition of the initial rate of Ca²⁺ transport in mussel mitochondria compared to approximately 1.0 nmole La³⁺ mg protein^?1 in rat liver mitochondria. It is concluded that mussel mitochondria possess a comparable Ca²⁺ transporter to vertebrate mitochondria and possible reasons for resistance to La³⁺ are discussed.     

Isolated and combined impacts of blue mussels (Mytilus edulis) and barnacles (Balanus improvisus) on structure and diversity of a fouling community

Effects of two presumably dominant competitors, the blue mussel Mytilus edulis and the barnacle Balanus improvisus on recruitment, population dynamics and community structure on hard substrata were experimentally investigated in the subtidal Kiel Fjord, Western Baltic. The hypothesis that blue mussels and/or barnacles are local dominants and strongly influence succession and community structure was tested by monitoring succession in the presence and absence of simulated predation on either or both species. Manipulations included blue mussel removal, barnacle removal, combined blue mussel and barnacle removal, as well as a control treatment for natural (non-manipulated) succession. In the second part of the experiment, recovery from the treatments was monitored over 1 year.During the manipulative phase of the experiment, blue mussels had a negative effect on recruitment of species, whereas barnacles had no significant effect. Even so, a negative synergistic effect of blue mussels and barnacles was detected. Calculation of species richness and diversity H′ (Shannon Index) showed a negative synergistic effect of blue mussels and barnacles on community structure. Additionally, diversity H′ was negatively affected by the dominant competitor M. edulis. These effects were also detectable in the ANOSIM-Analysis. The non-manipulative phase of the experiment brought about a drastic loss of diversity and species richness. Blue mussels dominated all four communities. Barnacles were the only other species still being able to coexist with mussels. Effects of simulated predation disappeared fast.Thus, in the absence of predation on blue mussels, M. edulis within a few months dominates available space, and diversity of the benthic community is low. In contrast, when mussel dominance is controlled by specific predators, more species may persist and diversity remains high.