Ecophysiological importance of the isolation response of hibernating blue mussels (Mytilus edulis)

Blue mussels, Mytilus edulis L., were collected at a site in Trondheimsfjordon, Norway in 1980 and 1981. The mussels isolate themselves from the ambient seawater for long periods in winter. When isolated, the mussels maintain a high osmotic concentration in the mantle fluid as the external salinity drops. The mussels display anaerobic metabolism when they are isolated. The isolation response is likely to be favourable to the mussels, with regard both to the energy budget and the freezing tolerance.     

Uptake and depuration of organic contaminants by blue mussels (Mytilus edulis) exposed to environmentally contaminated sediment

Experiments were designed to expose blue mussels (Mytilus edulis) to contaminated sediment collected from Narragansett Bay, Rhode Island, USA in 1982. Measurements were taken to allow comparisons of the uptake and depuration of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs). In addition, concentration factors in the mussels were calculated separately against the dissolved and particulate phase concentrations and the results from the exposure and control systems were compared. Both PAHs and PCBs were rapidly accumulated by the mussels exposed to the contaminated sediment. After the mussels were transferred to control seawater, individual PAHs were depurated with half-lives ranging from 12 to 30 d. Individual PCBs showed depuration half-lives which ranged from 16 to 46 d. Concentration factors in the mussels calculated against the particulate phase concentrations were very different in the exposure and control systems. Concentration factors calculated using only the dissolved phase concentrations (bioconcentration factors) showed excellent agreement in the two systems, possibly an indication that the dissolved phase was the direct source of the contaminants accumulated by the mussels. The bioconcentration factors for PCBs were higher than those of PAHs when compounds with similar n-octanol/water partition coefficients were compared.     

Ice nucleation in the blue mussel (Mytilus edulis)

The ice nucleation temperatures of different homogenised organs of the mussel Mytilus edulis L. were examined. The stomach and the hemolymph had the highest nucleation temperatures. In the homogenised stomach the nucleation temperatures were fairly constant throughout the year, whereas the nucleation temperatures of the hemolymph increased in the cold season. Bacterial growth experiments, microfiltration, and experiments using antibiotics indicated that the nucleators were not of bacterial origin. The nucleation temperatures of natural seawater were approximately −9 °C, whereas seawater made from distilled water and sea salt had nucleation temperatures of about −17 °C. The nucleation temperatures of the seawater were reduced when the seawater was filtered by the mussels. However, no clear indication that the nucleators in the stomach were obtained from the seawater was found. Stomach homogenates from mussels kept in nucleator-free water had the same supercooling points as stomach homogenates from mussels kept in natural seawater. This indicates that the nucleators in the stomach are not obtained from the seawater. The temperature and light conditions examined in the present study did not significantly influence the hemolymph ice nucleation temperatures of mussels kept in the laboratory. Received: 24 October 1996 / Accepted: 16 November 1996     

Shell growth response of mussels (Mytilus edulis) exposed to toxic microalgae

Mussels (Mytilus edulis) were exposed to the algaeAlexandrium ostenfeldii, Chrysochromulina polylepis, Gyrodinium aureolum, Gymnodinium galatheanum andHeterosigma akashiwo for 24 h; significant reductions in growth rate, as compared to the control, were observed after exposure toA. ostenfeldii, C. polylepis, G. aureolum andG. galatheanum at initial concentrations of 4.5 × 10⁶, 110 × 10⁶, 9 × 10⁶ and 120 × 10⁶ cells l^–1, respectively. Exposure to high initial concentrations of the non-toxic algaeTetraselmis suecica (174 × 10⁶ cells l^–1) andIsochrysis galbana (610 × 10⁶ cells l^–1) showed no adverse effect on growth rate. When mussels with reduced growth were transferred to clean seawater, they recovered to > 90% of control growth within 2 to 4 d. Exposure to algal filtrates of the toxic algal cultures produced no reduction in growth rate.     

Labial palps of the blue mussel Mytilus edulis (Bivalvia: Mytilidae)

As a basis for understanding the functions of labial palps in the blue mussel Mytilus edulis, the structure and histology of palps were studied using light and scanning electron microscopy. Mussels used in the present study were collected in August 1993 and April 1994. The palp ridged surface is characterized by the presence of a smooth but densely ciliated dorsal fold, upon which rests the corresponding demibranch ventral region. The underside of the dorsal fold and the palp ridges fuse to form vestigial ciliated tracts. The dorsal fold is capable of contraction, allowing it to cover variable amounts of the ridged surface. Two different types of mucocyte are present on the palp ridged surface: subepithelial, glandular, acid-dominant secretion mucocytes and epithelial mucocytes characterized by neutral secretions. In histological section, these mucocytes appear to be concentrated on anatomical features known to intervene in particle handling. The anatomical and histological features of the smooth surface are typical of bivalve labial palps, except that the dense ciliation of the dorsal fold begins in the dorsal region of the smooth surface, indicating the possible origin of this feature. Previous studies on M. edulis point to the palps as the probable site for both ingestion volume control and particle selection; the anatomical basis of the present study should facilitate further research on these aspects.     

Blue mussels, Mytilus edulis, at the edge of the range: population structure, growth and biomass along a salinity gradient in the north-eastern Baltic Sea

The blue mussel, Mytilus edulis, is the most conspicuous animal species in the northern Baltic rocky sublittoral. In the studied area the species lives at the margin of its salinity tolerance. Although dwarfed by the low-salinity conditions, blue mussels in the northern Baltic are very abundant and have a decisive role in the benthic and pelagic ecosystems. We studied abundance, size distribution, biomass and growth rate of blue mussels along a 270 km salinity gradient in the northern Baltic Sea. Samples (n=317, 1998-1999) from moderately exposed and exposed rocky shores at seven study areas were collected in the southern Archipelago Sea in the west and into the central Gulf of Finland in the east, where the species is becoming increasingly rare. The results show a marked decline in mean mussel size and biomass from the saline west to the less saline east. The growth rate also decreased with lower ambient salinity. However, abundance of small mussels was considerably higher in the central and eastern parts of the study area. Vertically, the highest biomass was recorded at intermediate depths (5 and 8 m), being lower at both shallower (3 m) and deeper bottoms (12 m). It is concluded that salinity is the foremost factor determining size structure and growth rate among populations within the area. The results suggest that predation further influences the population structure of blue mussels living at the edge of their range in the central Gulf of Finland ultimately set by their salinity tolerance.     

Ecological gradients and relative abundance of native (Mytilus trossulus) and invasive (Mytilus galloprovincialis) blue mussels in the California hybrid zone

Marine communities are experiencing unprecedented rates of species homogenization due to the increasing success of invasive species, but little is known about the mechanisms that allow a species to invade and persist in a new habitat. In central California, native (Mytilus trossulus Gould 1850) and invasive (Mytilus galloprovincialis Lamarck 1819) blue mussels and their hybrids co-exist, providing an opportunity to analyze the mechanisms that determine the distributions of these taxa. Spatial and temporal variation in temperature and salinity and the relative frequencies of these mussel taxa were examined between 2000 and 2004 at four sites in San Francisco Bay and four in Monterey Bay, which were chosen for their different positions along inferred estuarine/oceanic gradients in the hybrid zone. Mussels were genetically identified as the parent species or hybrids by amplifying regions of two species-specific loci: the adhesive byssal thread protein (Glu-5′) and the internal transcribed spacer region of ribosomal DNA (ITS 1). The proportion of M. trossulus at the eight hybrid zone sites correlated negatively with average salinity (R ²=0.60) and positively with maximal temperature (R ²≥0.72), a somewhat unexpected result given what is known about the phylogeography of this species. The proportion of M. galloprovincialis showed the opposite pattern. The proportion of hybrids was correlated neither with habitat temperature nor salinity. Genotypes of mussel populations at an additional 13 sites from Coos Bay, Oregon (latitude 43.35°N) to Long Beach, California (latitude 33.72°N), sampled at various intervals between 2000 and 2004, were also determined. This survey confirmed previous reports that the hybrid zone lies between Monterey and the Cape Mendocino region (latitudes 36.63°N–40.5°N). Within Monterey and San Francisco Bays, however, the temporal comparisons (1990s vs. 2000s) revealed abrupt changes in the proportions of the two parent species and their hybrids on annual and decadal time scales. These changes indicate that the blue mussel populations are in a highly dynamic state. The survey also showed that, regardless of habitat, M. trossulus is consistently of smaller average size than either M. galloprovincialis or hybrids.

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Genetic population structure of a species' complex of blue mussels (Mytilus spp.)

Blue mussels representing two nominal species (Mytilus trossulus Gould, 1850 and Mytilus galloprovincialis Lamarck, 1819 were collected from 28 intertidal locations along the Pacific coast of the USA in 1990–1991 (total N=1255) and examined for variation at 15 allozyme loci. Twelve samples, mostly from a region of suspected hybridization, were analyzed for variation in seven shell characters. Principal-components analysis of allozyme data revealed three groups based on first principal-component scores, which were identified as M. trossulus, M. galloprovincialis, and hybrids. Canonical discriminant analysis of shell characters was less successful in separating mussels into discrete groups. Each location was characterized for four environmental variables: (1) temperature, (2) salinity, (3) tidal height and (4) degree of exposure to wave action, which were then used as independent variables in a series of multiple-regression analyses, with the proportions of the two species as dependent variables. Temperature and salinity had significant (P<0.05) effects on the macrogeographic distribution of the two species, whereas the effects of height in the tidal zone and degree of wave exposure were not statistically significant. Salinity was found to have a greater influence than temperature on the microgeographic distribution of the two species. M. trossulus was more abundant at locations with lower temperatures and greater salinity variation than M. galloprovincialis. The two species appear to be ecologically distinct, and their genetic integrity is at least partly the result of environmental heterogeneity.     

Biochemical genetic variation at a leucine aminopeptidase (LAP) locus in blue (Mytilus galloprovincialis) and greenshell (Perna canaliculus) mussel populations along a salinity gradient

In several marine bivalve species, biochemical genetic variation at a leucine aminopeptidase (LAP) locus is associated with environmental variability, primarily salinity fluctuation. Population genetic variation at a LAP locus was investigated here in two sympatric mussel species (Mytilus galloprovincialis and Perna canaliculus) from three locations along a salinity gradient in Wellington Harbour, New Zealand. The data for M. galloprovincialis and P. canaliculus do not support the hypothesis that the LAP polymorphism in either species is associated with salinity variation among adult mussels. Due to the absence of small mussels among the samples it is not possible to discount the hypothesis that selection acts primarily against juveniles, as it does for M. edulis in Long Island Sound, USA. Wellington Harbour populations of M. galloprovincialis exhibited large and often highly significant heterozygote excesses at the LAP locus, whereas populations of P. canaliculus from the same locations exhibited large and highly significant heterozygote deficiencies. The reason for this inter-specific difference in population structure is unknown. If it is the result of selection, this suggests that selection acts differentially upon the two species, because demographic attributes and reproductive biology are very similar in the two species. For both M. galloprovincialis and P. canaliculus, significant levels of population genetic heterogeneity were recorded among three locations separated by only 8 to 12 km. Neither species exhibited shell length-dependent genetic variation at the LAP locus, suggesting that for these two species the LAP polymorphism is not associated with variation in shell length. Received: 30 December 1996 / Accepted: 6 January 1997     

Size-dependent, spatial and temporal genetic variation at a leucine aminopeptidase (LAP) locus among blue mussel (Mytilus galloprovincialis) populations along a salinity gradient

Biochemical genetic variation at a leucine aminopeptidase (LAP) locus is related to salinity variation in several marine bivalve molluscs. This paper details an investigation of the Long Island Sound model of LAP selection (LAP genotype-dependent mortality occurring among newly settled Mytilus edulis mussels) carried out in 1997 among three M. galloprovincialis mussel populations along the salinity gradient of Wellington Harbour, New Zealand. Significant LAP genotypic heterogeneity was observed at the LAP locus between small (<25 mm shell length) and large (>25 mm shell length) M. galloprovincialis from Petone and Eastbourne (the two sites experiencing the greatest salinity variation), whereas genotypic heterogeneity was not significantly different between small and large mussels from Seatoun (the site experiencing the least salinity variation). The Lap ³ allele decreased in frequency and the Lap ⁴ allele increased in frequency at Petone and Eastbourne, whereas the Lap ³ and Lap ⁴ allele frequencies remained effectively constant at Seatoun. Both these findings are consistent with the Long Island Sound model of selection. At all three locations, the Lap ^3,3 genotype decreased in frequency from small to large mussels, whereas the Lap ^3,4 genotype increased in frequency from small to large mussels. All other LAP genotypes occurred at low frequencies (<0.10) at all three locations and showed no evidence of frequency change from small to large-size mussels nor evidence of clinal change among the three locations. These genotype frequency data possibly indicate that the Lap ^3,3 genotype is at a selective disadvantage compared to the Lap ^3,4 genotype at all three locations, and that this selective disadvantage is related to the extent of salinity variation which exists at each location. Further investigation is required before it can be determined if the Long Island Sound model of selection best describes the size-dependent and location-specific changes in LAP allele and genotype frequencies along this salinity gradient. Comparison of the population genetic structure at the LAP locus in 1995 and in 1997 revealed a profound change from heterozygote excesses to heterozygote deficiencies for all three M. galloprovincialis populations. The reason for the change is unknown, but the change indicates that population genetic structure at the LAP locus is highly variable in time, but consistent in space, among these M.␣galloprovincialis populations. Received: 5 February 1998 / Accepted: 27 May 1998     

Role of the kidney in determining the whole soft tissue zinc concentration of individual mussels (Mytilus edulis)

Mussels (Mytilus edulis) were collected from an unpolluted site in Newfoundland in 1984 and 1985 in order to determine the zinc levels of the whole soft tissues and individual organs. Individuals were collected in such a way as to eliminate all known physiological and environmental factors which might have led to differences in zinc concentration between individual mussels. The inherent variability of the whole soft tissue zinc concentration was found to be very high with a coefficient of variation (CV) around 40%. Most individual organs had relatively low mean zinc concentrations with small values for CV (10–18%). However, the kidney showed a relatively high mean zinc concentration (828 ppm) with a remarkable degree of variability (94–3 410 ppm) (CV=78%) and is responsible not only for the high degree of inherent variability in the population but also for the high levels of zinc found in the whole soft tissue of some individual mussels. Those individuals with high whole soft tissue zinc concentrations often had high kidney zinc concentrations and stored as much as 53% of their zinc load in the kidney (which accounted for only 1–3% of the total weight). However, other individual mussels with low whole soft tissue zinc concentrations had low kidney zinc concentrations and stored as little as 3% of their body zinc load in the kidney. Hence, the role that the kidney plays in zinc storage varies markedly from mussel to mussel.M.S.R.L. Contribution No. 624     

Genotoxic effects of Ag2S and CdS nanoparticles in blue mussel (Mytilus edulis) haemocytes

The use of functionalised metal sulphide nanoparticles (NPs) for nanoremediation and biomedical application is rapidly increasing, which could lead to significant inputs into the marine environment. The potential impact of some NPs on marine organisms is still poorly understood. In the present paper the genotoxic potential of Ag₂S and CdS NPs on Mytilus edulis haemocytes was assessed. MPEG-SH (thiol-terminated methyl polyethylene glycol), was used as capping agent to avoid NPs agglomeration. TEM analysis showed that the Ag₂S NPs size was 13±7 nm, whereas CdS quantum dots had an average diameter of 4±1 nm. DNA integrity was evaluated by Comet assay following exposure to increasing concentration series (0.01–10 mg/L). Both silver and cadmium NPs showed genotoxic effects at the highest dose. MPEG-SH was also found to exert a weak genotoxic activity, suggesting that at least part of the genotoxic potential of functionalised NPs on mussel cells might be attributable to the capping agent. These results confirm the genotoxic potential of Ag₂S NPs for mussel cells and demonstrated, for the first time, that CdS NPs is genotoxic in a marine organism.     

A proteomic approach to the study of the marine mussels Mytilus edulis and M. galloprovincialis

In this study, a proteomic approach was applied for the generation of reference maps and subsequently to detect, quantify and compare the global protein expression between two related species of marine mussels, Mytilus edulis and Mytilus galloprovincialis, growing in their own geographical habitats. A comparative study of the protein profiles generated from analytical two-dimensional electrophoresis gels was performed, and changes in protein expression were analysed quantitatively by computer analysis. An average of 1,278 spots per gel was detected in 16 individuals (8 M. edulis and 8 M. galloprovincialis); however, not all spots were included in the study. Expression of 420 spots was compared, and significant differences in the intensity levels were detected in 37 protein spots (8.8%). Fifteen proteins showed higher expression in M. edulis, and 22 proteins, in M. galloprovincialis. The technique of peptide mass fingerprinting using MALDI-TOF (matrix-assisted laser desorption ionisation/time-of-flight) and/or nanoelectrospray double subfragmentation mass spectrometry enabled the unambiguous identification of 15 of these 37 differentially expressed proteins. Most of the identified proteins can be grouped basically into four broad functional classes: cytoskeletal and myofibrillar proteins, proteins associated with stress response, proteins associated with the storage or production of energy, and proteins related to rearrangement in the synthesis of native structures. These results expand our understanding of the molecular differentiation of the two mussel taxa and serve as a useful base for future ecological, physiological and genetic studies.     

Influence of body size on the uptake,depuration, and bioaccumulation of polychlorinated biphenyl congeners by Baltic Sea blue mussels,Mytilus edulis

The present study was designed to examine the influence of body size on the bioaccumulation of polychlorinated biphenyl (PCB) congeners by Baltic Sea blue mussels, Mytilus edulis L. This was done, firstly, by establishing the relationship (as a power function: PCB tissue conc = a tissue dry wt^b) between tissue concentration and body weight for seven PCB congeners (International Union of Pure and Applied Chemistry No. 52, 101, 105, 118, 138, 153, and 180) in field sampled mussels; and, secondly, by assessing the influence of body weight on the uptake clearance coefficients, the depuration rate coefficients and the calculated bioaccumulation factors (BAFs) of three ¹⁴C-labelled PCB congeners (IUPAC No. 31, 49, 153) in mechanistic kinetic experiments. Both the background tissue concentrations of PCB 138, PCB 153 and PCB 180 and the predicted BAF values in the kinetic experiments correlated negatively with body weight (b=-0.17 and-0.31, respectively). Of the two kinetic rate coefficients examined, only the uptake clearance rate showed weight dependency (b=-0.32, i.e. negative correlation with body weight), whereas depuration rates were unaffectd by body weight. Uptake clearance rates and BAFs adjusted for body weight increased with the octanol/water partition coefficient (K_ow) of the congener, whereas depuration rates dectreased with K_ow. These observations suggest that size-dependent bioaccumulation of hydrophobic organic contaminants (HOCs) in suspension-feeding bivalves is driven by size-related differences in uptake rate, although several other mechanisms may also affect bioaccumulation in natural mussel beds (e.g. sizerelated differences in lipid content, production, and contaminant exposure). From an ecotoxicological perspective size-dependent bioaccumulation implies not only that variability due to body size differences has to be dealt with in experimental designs, but also that several ecological factors such as size-specific predation and shifts in population structure may affect HOC cycling by dense populations of bivalve suspensionfeeders.     

The effect of water temperature on drilling and ingestion rates of the dogwhelk Nucella lapillus feeding on Mytilus edulis mussels in the laboratory

In highly seasonal intertidal habitats, changes in temperature through the year may drive substantial shifts in feeding and growth rates of organisms. For the dogwhelk Nucella lapillus, attacking and consuming Mytilus edulis mussels can take hours or days, depending on temperature. Handling time of dogwhelks feeding on mussels is therefore greatly affected by ocean temperature. I recorded attack time in the laboratory, partitioned into drilling and consumption time, for juvenile dogwhelks across a range of seawater temperatures representative of field seawater temperatures during the main growing seasons of summer and autumn. The combined length of a drilling attack and subsequent ingestion time tripled across the 10 °C decline in water temperatures from July through November, driven primarily by an increase in ingestion time. The observed reduction in handling time, coupled with projected sea surface warming in New England by the end of the twenty-first century, could extend the length of the growing season for Nucella and subsequently have cascading effects on the prey community.     

Introgression and mitochondrial DNA heteroplasmy in the Baltic populations of mussels Mytilus trossulus and M. edulis

A strong clinal change in salinity occurs between the Baltic Sea and the North Sea, Atlantic Ocean, in the Danish Straits, where hybridization zone between mussels Mytilus edulis and M. trossulus has been reported. Eleven samples of mussels were studied from the Danish Straits and the inner Baltic Sea. Extensive introgression of M. edulis alleles from the North Sea into populations throughout the Baltic was ascertained for mitochondrial DNA (mtDNA) and two nuclear markers (ME15–16 and ITS). In the opposite direction, introgression of M. trossulus alleles into the M. edulis background was observed at the EFbis nuclear marker in populations from Kattegat (Danish Straits). While only M. edulis F (female) mtDNA was present in the Baltic, there were still strong differences in frequencies in the control region length variants between the Danish Straits and the inner Baltic samples, and weaker variation in coding region ND2–COIII haplotype frequencies. In the assays of the two mtDNA regions, various patterns of heteroplasmy were detected in 32% of all the studied individual mussels; this includes the presence of distinct, independently inherited M and F mitochondria in males, as well as the presence of two different distinguishable F genomes. The male-inherited M mtDNA genomes are quite common in the mussels from the Danish Straits, but very rare in males from the inner Baltic. Instead, a recombined control region variant (1r), which seems to have taken over the role of the M genome, was present in a number of specimens in the Baltic. Observations of heteroplasmy for two F genomes in some females and males confirm disruptions of the doubly uniparental inheritance mechanism in the hybrid Baltic Mytilus.     

Hybridization between the blue mussels Mytilus galloprovincialis and M. trossulus along the Pacific coast of North America: evidence for limited introgression

Two species of blue mussel, Mytilus galloprovincialis and M. trossulus, co-occur and hybridize along the Pacific coast of North America. Using a set of polymerase chain-reaction (PCR)-based genetic markers which diagnostically identify these species, we show that they are sympatric from the Cape Mendocino region to the Monterey Peninsula in northern and central California, USA. Mussels with hybrid genotypes were detected in all populations sampled in the region of sympatry, and the frequency of hybrid genotypes in individual hybrid populations ranged from 13 to 44%. Significant frequencies of first-generation backcross genotypes were detected in two individual hybrid zone populations (Berkeley and Monterey Marina) and in the hybrid zone as a whole, indicating that the potential exists for introgression between M. galloprovincialis and M. trossulus. Despite this potential, we found no evidence of advanced introgression beyond first-generation backcrosses, suggesting that gene flow between M. galloprovincialis and M. trossulus has been quite limited. The frequency of mussels with M. trossulus and hybrid genotypes declined abruptly south of Monterey Peninsula, while the frequency of mussels with M. galloprovincialis and hybrid genotypes declined precipitously north of Cape Mendocino. These abrupt genetic discontinuities indicate that this blue mussel hybrid zone is presently positioned between two prominent coastal features and there is little, if any, export of alleles from the hybrid zone into bordering parental populations. Received: 20 August 1997 / Accepted: 26 October 1998