AFLP-based genetic linkage maps of the blue mussel (Mytilus edulis)

We report the construction of the first genetic linkage map in the blue mussel, Mytilus edulis. AFLP markers were used in 86 full-sib progeny from a controlled pair mating, applying a double pseudo-test cross strategy. Thirty-six primer pairs generated 2354 peaks, of which 791 (33.6%) were polymorphic in the mapping family. Among those, 341 segregated through the female parent, 296 through the male parent (type 1:1) and 154 through both parents (type 3:1). Chi-square goodness-of-fit tests revealed that 71% and 73% of type 1:1 and 3:1 markers respectively segregated according to Mendelian inheritance. Sex-specific linkage maps were built with mapmaker 3.0 software. The female framework map consisted of 121 markers ordered into 14 linkage groups, spanning 862.8 cM, with an average marker spacing of 8.0 cM. The male framework map consisted of 116 markers ordered into 14 linkage groups, spanning 825.2 cM, with an average marker spacing of 8.09 cM. Genome coverage was estimated to be 76.7% and 75.9% for the female and male framework maps respectively, rising to 85.8% (female) and 86.2% (male) when associated markers were included. Twelve probable homologous linkage group pairs were identified and a consensus map was built for nine of these homologous pairs based on multiple and parallel linkages of 3:1 markers, spanning 816 cM, with joinmap 4.0 software.     

Genetic studies of laboratory reared mussels, Mytilus edulis: heterozygote deficiencies, heterozygosity and growth

Offspring derived from several single pair matings and a mass mating of Mytilus edulis were cultured up to the juvenile stage in the laboratory. Samples were collected from the cultures at an early post- settlement ('spat') stage and again at the juvenile stage and were scored at up to nine enzyme loci using cellulose acetate and starch gel electrophoresis. An overall significant deficiency of heterozygotes was evident at the juvenile stage but not at the spat stage. Selection against heterozygotes (underdominance) is proposed as the cause.
For each mating, shell lengths of juvenile mussels were measured and individuals were classed according to the number of loci at which they were heterozygous. No significant correlations were evident between heterozygosity class and either growth (shell length), or coefficient of variation of shell length. It is argued that these results provide support for the 'associative overdominance' rather than the 'overdominance' hypothesis as an explanation for heterozygosity/growth correlations in marine bivalves.
These data also show that although heterozygosity and growth co-occur in most data sets from natural populations, they may have distinctly differing causes.     

Introgression patterns in the mosaic hybrid zone between Mytilus edulis and M. galloprovincialis

Hybrid zones are fascinating systems to investigate the structure of genetic barriers. Marine hybrid zones deserve more investigation because of the generally high dispersion potential of planktonic larvae which allows migration on scales unrivalled by terrestrial species. Here we analyse the genetic structure of the mosaic hybrid zone between the marine mussels Mytilus edulis and M. galloprovincialis, using three length-polymorphic PCR loci as neutral and diagnostic markers on 32 samples along the Atlantic coast of Europe. Instead of a single genetic gradient from M. galloprovincialis on the Iberian Peninsula to M. edulis populations in the North Sea, three successive transitions were observed in France. From South to North, the frequency of alleles typical of M. galloprovincialis first decreases in the southern Bay of Biscay, remains low in Charente, then increases in South Brittany, remains high in most of Brittany, and finally decreases again in South Normandy. The two enclosed patches observed in the midst of the mosaic hybrid zone in Charente and Brittany, although predominantly M. edulis-like and M. galloprovincialis-like, respectively, are genetically original in two respects. First, considering only the various alleles typical of one species, the patches show differentiated frequencies compared to the reference external populations. Second, each patch is partly introgressed by alleles of the other species. When introgression is taken into account, linkage disequilibria appear close to their maximum possible values, indicating a strong genetic barrier within all transition zones. Some pre- or postzygotic isolation mechanisms (habitat specialization, spawning asynchrony, assortative fertilization and hybrid depression) have been documented in previous studies, although their relative importance remains to be evaluated. We also provided evidence for a recent migratory 'short-cut' connecting M. edulis-like populations of the Charente patch to an external M. edulis population in Normandy and thought to reflect artificial transfer of spat for aquaculture.     

双酶水解紫贻贝制备酵母调味料研究

利用复合蛋白酶、复合风味蛋白酶、中性蛋白酶对紫贻贝进行双酶水解,水解效果比较后选择使用复合蛋白酶和复合风味蛋白酶作为复合水解酶,同时采用产酯酵母发酵技术制备调味料,通过电位滴定法测定单菌株和多菌株发酵对双酶水解贻贝肉产总酯的影响。结果表明：产酯酵母1274在双酶水解后,接种量为5％,发酵温度28℃,发酵时间72h时总酯含量为0．65％,相同条件下,产酯酵母1274和1202多菌株发酵时总酯含量为0．78％,经产酯酵母发酵后的调味料,酯香味浓郁,给予产品以发酵特有的风味。     

Growth and longevity of Mytilus edulis (L.) from northeast Europe

The growth of mussels, Mytilus edulis (L.), was studied in most of the northeastern part of their distribution. The growth, longevity and maximal size of molluscs from 24 wild populations and one cultured population located in the White Sea and the southeast part of the Barents Sea were compared. The 25 studied populations were combined in six clusters. The maximal longevity and the size of the mussels varied between 7 and 18 years and 25.5 and 77.7 mm, respectively. The geographical location of the population within the studied region did not affect either maximal longevity or maximal size, or the growth rate of mussels. However, these parameters were influenced by local habitat conditions, primarily connected with the position within intertidal and subtidal zones. Animals inhabiting the intertidal zone were characterized by relatively low growth performance, a short life span and a small size. The longest life span was typical for deep subtidal mussels, whereas the highest growth rate was recorded in the cultured population and in the upper subtidal habitats. Growth patterns of Mytilus edulis in the subarctic White and the Arctic southeast Barents seas are similar to those reported from other parts of the area of distribution. Therefore, growth was mostly determined by local environmental factors, including those related to vertical zonation, rather than by latitude/longitude and related temperature effects.     

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.     

Polygodial,a Potent Attachment-inhibiting Substance for the Blue Mussel,Mytilus edulis galloprovincialis from Tasmannia lanceolata

A highly potent attachment-inhibitor, polygodial, was isolated from a hexane extract of the leaves of Tasmannia lanceolata. The attachment-inhibiting activity of polygodial against the blue mussel was increased 4-fold when used in combination with sorbic acid, anethole, and indole.     

Cadmium and copper accumulation in the common mussel Mytilus edulis in the Western Scheldt estuary: a model approach

The Western Scheldt of the Dutch Delta area is severely contaminated with trace metals. Accumulation models of trace metals in the mussel Mytilus edulis are required to predict the biological efficiency of reductions in the metal and organic matter load. Two models are constructed: a black-box model and a physiologically structured model. The black-box model predicts metal accumulation in mussels from uptake and elimination parameters. The physiological model attempts to improve predictions by taking into account the kinetics of individual uptake and elimination routes. These in turn, are taken as depending upon two more general physiological processes, the ventilation rate and the metabolic rate. Metal uptake via food and water are expressed as relative fractions. Metal input is differentiated into particulate adsorbed, and dissolved species.The reliability of the two models is evaluated by comparing predicted concentrations for mussels with measurements. Model predictions for copper deviate less than 100% from measured concentrations, but neither model appears to predict cadmium concentration with sufficient accuracy since deviations of more than 100% occured. The introduction of physiological refinements did not improve performance. Food mediated contributions for cadmium and copper to total body burden had been overestimated in the model by a factor of 100 when compared to literature values. The physiological model did predict that the ratio of food mediated contribution to total body burden is probably different for cadmium and copper and decreases with increasing salinity for both. As yet there are no measurements available to confirm such predictions.We conclude that additional laboratory experiments should be done for a better understanding of why there is poor agreement between the few field observations and the simulations. In these experiments mussels grown under different environmental condition can be tested for their accumulation capacity of trace metals. More field observations are needed.     

Genetic divergence and a hybrid zone between Baltic and North Sea Mytilus populations (Mytilidae: Mollusca)

Populations of the common mussel ( Mytilus edulis ) from the North Sea area (Skakerrak-Kattegat) and those from the Baltic Sea are almost diagnostically differentiated at five out of 22 studied allozyme loci; at a further seven loci, alleles predominant or common in one area are nearly absent in the other. Genetic distance was estimated at 0.28; this is similar to the distances of these populations to the Mediterranean mussel M. galloprovincialis. The three mussel types obviously represent equal evolutionary divergence from one another, and should also be taxonomically equally separated; a semispecies rank within a more comprehensive M. edulis complex or superspecies is suggested. The age of the Baltic mussel type ( 'M. trossulus' ), as an independent evolutionary lineage, is probably far greater than that of the post-glacial Baltic Sea.
Allele frequencies change gradually and in parallel when entering from the Kattegat through the Sound into the Baltic. Only a slight Wahlund effect at the strongly diverged Gpi and Pgm loci was found in intermediate populations, indicating that extensive hybridization of the two taxa takes place in the area. However, strong interlocus genotypic associations suggest that selection against hybrids is intense in later generations; the c. 100 km wide hybrid zone is narrow relative to the dispersal distance. The genotypic structure of the Lap locus does not conform with those of the other loci studied in the hybrid zone; it cannot be viewed merely as a neutral marker of the process of hybridization.     

The ecophysiological response of mussels (Mytilus edulis) in mesocosms to a range of inorganic nutrient loads: simulations with the model EMMY

EMMY is an ecophysiological model of the growth and reproduction of a single mussel (Mytilus edulis L.). It contains feedback loops in the uptake and metabolism of food and in the partitioning of carbon and nitrogen to the internal state variables somatic tissue, storage, organic shell matrix and gametes. In this paper EMMY is used to simulate individual mussel growth in a series of mesocosm experiments with different inorganic nutrient loads (N and P). The experiments explore the impact of eutrophication reduction scenarios on mussel growth under defined and controlled conditions.In earlier studies EMMY was calibrated using expert knowledge on growth and reproduction during a period of 5 years. The resulting calibrated model was validated for system inputs and observations of three ecosystems with significantly different food and silt concentrations. EMMY reproduced the mussel growth sufficiently accurate in ecosystems with moderate or high food concentrations. In this study EMMY was adapted in order to cope with low food concentrations, then recalibrated (using the original calibration data and procedure) and applied without further calibration to 3 replicated mesocosm experiments. The EMMY simulations in this study show the ecophysiological response of mussels to different food (phytoplankton and detritus) concentrations. It is concluded that the mussels can adapt to significantly reduced food concentrations, due to inorganic nutrient load reduction, and still maintain growth.     

Differential effects of the severe winter of 1995/96 on the intertidal bivalves Mytilus edulis, Cerastoderma edule and Mya arenaria in the Northern Wadden Sea

Intertidal mussel beds were severely damaged by scouring ice floes during the winter of 1995/96. Aerial surveys before and after the winter showed that more clusters of mussel beds vanished in a region with a higher areal share of tidal flats and a lower salinity, suggesting that the amount of ice present determined the magnitude of the disturbance on beds of Mytilus edulis. Nehls and Thiel [(1993) Neth J Sea Res 31:181–187] observed a strikingly similar spatial pattern of disturbances caused by severe storms in the Wadden Sea. Areas on mussel beds mechanically undisturbed by ice showed no reduced abundance and biomass of mussels, indicating that temperature alone was of little importance as a lethal factor. Conversely, Cerastoderma edule was strongly affected by low temperature. On average 80% died during the winter with extinctions up to 100% in the high tidal zone. At the lowest tidal level, surviving cockles were larger than those killed by the frost. A reinvestigation of sampled sites in autumn revealed that substantial further mortality had occurred during spring and summer which may constitute a time-lag effect of the preceding winter. There was no increased mortality in juvenile and adult Mya arenaria during the winter of 1995/96, confirming that this clam is a hard-winter species like Macoma balthica. Received in revised form: 7 May 2001 Electronic Publication     

The physiological ecology of two populations of Mytilus edulis L.

Summary Seasonal cycles in the rates of oxygen consumption, feeding, absorption efficiency and ammonia-nitrogen excretion in two populations of Mytilus edulis were measured in the field under ambient conditions and related to body size, the gametogenic cycle, the concentration of suspended particulate matter in the water and temperature. Relationships between the various physiological variables are also considered and protein and energy budgets estimated. Both the scope for growth and the relative maintenance cost were seasonally variable, demonstrating a minimum capacity for growth in the winter and a maximum capacity in the summer. In one population subjected to abnormally high temperatures in the winter the scope for growth was negative for four or five months between January and May. These population differences are discussed and the potential for using physiological integrations in intra-specific comparisons of fitness is identified.     

Regulation of particle transport within the ventral groove of the mussel (Mytilus edulis) gill in response to environmental conditions

To thoroughly understand the feeding processes of the blue mussel, Mytilus edulis, under the variable environmental conditions it experiences in nature, it is important to examine individually the different components of its feeding system. The ciliated ventral food groove represents one of these components, within which the majority of food particles trapped by the gill are transported to the labial palps and mouth. Any ability of the mussel to adjust food transport rates within this groove could serve as an important feeding regulatory mechanism in response to variations in the environment.

Mucous strand velocities in the ventral groove of the mussel, M. edulis, were determined by video endoscopy over different time periods and during short- and long-term manipulations of ambient particle concentration and temperature. Mucous strand velocity decreased with increasing ambient particle concentration at 14°C, but a similar relationship was not observed at 5°C. The data support the hypothesis that M. edulis possesses compensatory mechanisms to control particle transport at the level of the ventral groove cilia in response to changes in the environment. Furthermore, mucous strand velocity in the ventral groove increased when the ambient temperature of mussels acclimated to 5°C was increased to 15°C during acute and long-term acclimation temperature experiments. This response is consistent with standard physiological responses of ciliary systems to changes in temperature.     

Uptake and persistence of human associated Enterococcus in the mussel Mytilus edulis: relevance for faecal pollution source tracking

Aims:  Micro-organisms and molecular markers for microbial source tracking (MST) in coastal waters are often present at low numbers, and often exhibit significant variability in time and space. In this study, we investigated the uptake, accumulation, and persistence of human associated Enterococcus in the mussel Mytilus edulis .
Methods and Results:  The human associated molecular markers esp in Enterococcus faecium , and M66 in Enterococcus faecalis were targetted by PCR in seawater and mussel samples from coastal sites affected by sewage contamination. Both native mussels and mussels transplanted from pristine to polluted sites were included. The results showed that the esp and M66 markers were often not detectable in seawater whereas mussels were enriched in the markers. Human associated E. faecalis accumulated rapidly in M. edulis , and reached maximum levels after 4–6 h with concentration 30–300 times greater than in the surrounding seawater. Enterococcus faecalis retained in M. edulis showed a survival comparable to planktonic E. faecalis in seawater with half lives of 30 and 22 h, respectively. Human associated markers remained detectable for 120 h in M. edulis after faecal contamination.
Conclusions:  The study demonstrated that native and transplanted M. edulis can accumulate and retain human associated molecular markers relevant for MST.
Significance and Impact of the Study:  Mussels should be considered as additional targets in MST studies in coastal waters.     

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.     

Carbonic anhydrase activity and biomineralization process in embryos, larvae and adult blue mussels Mytilus edulis L.

To decide whether a physiological role can be attributed to enzymatic activity with respect to crystal formation and biomineralization of the first larval shell, carbonic anhydrase (CA) activity was measured in embryos and larvae of the blue mussels Mytilus edulis L. Also, CA activity was determined in the mantle edge and gonads of adult mussels with different shell length and condition index. The intention was to find a possible correlation between CA activity and adult shell calcification, i.e. gonadal maturation. The comparison of CA activity in different developmental stages of mussels and the results of an X-ray diffraction study of biomineralization processes in embryonic and larval shells indicate that CA activity is maximal at the end of several developmental stages. Consequently, the increase in CA activity precedes some physiological changes, i.e. the somatoblast 2d formation and the occurrence of the first calcite and quartz crystals in embryos, shell field formation in the gastrula stage, shell gland and periostracum production in trochophores, and rapid aragonite deposition in larval prodissoconch I and prodissoconch II shells. Furthermore, it was found that in adult mussels CA activity was quite variable and that in the mantle edge it was frequently inversely related to the activity in the gonad. Received: 28 November 1998 / Received in revised form: 30 August 1999 / Accepted: 31 August 1999     

Oxygen consumption,ammonia excretion,and filtration rate of the marine bivalve Mytilus edulis exposed to methamidophos and omethoate

In this study, the oxygen consumption, ammonia excretion, and filtration rate were monitored in Mytilus edulis in response to administration of the two pesticides, methamidophos and omethoate. Five sublethal concentrations (1, 10, 50, 100, and 200 µgL^?1) were administered over 96 h. Oxygen consumption rates increased following administration of all concentrations of methamidophos for 96 h and to the lower concentrations of omethoate (1, 10, and 50 µg L^?1) from 6 to 24 h. Over 24 h, oxygen consumption decreased significantly. Ammonia excretion rates were higher than the control after 36 h of exposure to methamidophos, while reduction was observed when M. edulis was exposed to omethoate. The O:N ratios increased when the animals were first exposed to these two pesticides and then decreased at all the concentrations. At the concentrations of 100 and 200 µgL^?1, the O:N ratio was below 30 after 72 h. The filtration rate of M. edulis decreased with the increasing concentration of pesticide exposure.     

Microbial biomass and activity in the vicinity of a mussel bed built up by the blue mussel Mytilus edulis

Our study focuses on the impact of the biosedimentary system mussel bed (Mytilus edulis) on the spatial variability of benthic microbial biomass and activity in relation to organic deposits. We sampled a transect of six stations from the muddy mussel bed towards a reference station in the sandflat in monthly intervals for 1 year. The sediment grain size fraction <63 μm and the total organic carbon (TOC) concentration decreased significantly towards the sandflat. Bacterial numbers and total microbial biomass (total adenylates) showed a high spatial variability and were not correlated to increasing distance from the mussel bed. However, a significant relationship with the TOC concentration was found. In contrast, the energetic status (adenylate energy charge) of the microbial community in the mussel bed was significantly lower than in the sandflat. A principal component analysis of the substrate-utilization pattern revealed clear differences between the microbial communities in the mussel bed and in the sandflat. Our results indicate that the sandflat may be dominated by a relatively specialized benthic microbial community with an increased efficiency in utilizing organic carbon sources. As a disadvantage, however, such r-strategists are only able to meet environmental changes within a comparatively narrow range. Benthic microbial communities in the vicinity of an M. edulis mussel bed, in contrast, are dominated by relative generalists with a greater physiological capacity to buffer discrete environmental changes. Such K-strategists show a lack of specialization which generally means a reduced efficiency in utilizing a particular resource. Received: 2 May 1999 / Received in revised form: 22 November 1999 / Accepted: 6 December 1999     

Aspects of the population genetics of Mytilus (Mytilidae; Mollusca) in the British Isles

Starch gel electrophoresis was used to study variation at 11 loci in mussels sampled mainly from British coastal sites. Two types of mussel were identified, Mytilus edulis, the common mussel and its southern relative Mytilus galloprovincialis. Several partially diagnostic loci were used to map the distribution of the two forms. Mytilus edulis was present at all sites sampled in Britain and Ireland but was at low frequency in SW England; M. galloprovincialis was detected in SW England, the south and west of Ireland. Scotland and NE England, but was absent from south Wales, the Irish sea coasts of Wales and Ireland, and SE England. Apart from the occurrence of M. galloprovincialis in NE England, this distribution conforms with the results of studies using morphological characters and parallels the distribution of many other southern species in Britain. At the microgeographical level, M. edulis was found to prefer more sheltered and estuarine conditions than M. galloprovincialis. Analysis using the best diagnostic loci showed that hybridization is occurring between M. edulis and M. galloprovincialis at all localities where they occur sympatrically but that the extent of hybridization varies considerably between localities. The distribution of localities having high proportions of hybrid individuals is best interpreted by assuming that hybrids have higher fitness than parental types at these localities. A study was made of variation within and between those localities where only M. edulis individuals were observed. Little significant geographic variation in allele frequency was detected, but significant deficits of heterozygotes compared with Hardy-Weinberg expectations were seen for most loci. Analysis suggests that the Wahlund effect is not involved and that the most likely cause of the deficit is low frequencies of null alleles. In M. edulis no differences in phenotypic variance in shell height and width were observed between samples of multiply heterozygous and multiply homozygous individuals and no genetic differences were found between juveniles and adults. Overall little evidence was found that balancing selection is responsible for maintenance of the polymorphisms studied in M. edulis. The pattern of geographic variation in gene frequencies in Mytilus in the British Isles is discussed in relation to variation in the south and north of Europe and North America. It is concluded that steep clines in gene frequencies in M. edulis observed by other workers in the Baltic and in Long Island Sound cannot be attributed to the presence of M. galloprovincialis.     

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.