Ultrastructural characterisation of Marteilia species (Paramyxea) from Ostrea edulis, Mytilus edulis and Mytilus galloprovincialis in Europe

A focused ultrastructural study of Marteilia spp. found in cultured Ostrea edulis, Mytilus edulis and Mytilus galloprovincialis from France and Spain was conducted with emphasis placed on haplosporosomes, striated plate-like inclusions and spore wall morphology. Two types of haplosporosome were identified, sphaeroid and oblate, which were common to the parasite in all 3 host species. A total of 492 haplosporosomes were measured; those from the Marteilia sp. in Mytilus spp. were marginally smaller than those in Ostrea edulis. Spore wall morphology was found to vary depending on the state of maturity of the parasite--the more mature the parasite, the thicker the wall surrounding it. It is suggested that the current criteria used to distinguish M. maurini from M. refringens are invalid and that M. maurini was relegated to a junior synonym of M. refringens.     

Electrophoretic investigation of systematic relationships in the marine mussels Modiolus modiolus L., Mytilus edulis L., and Mytilus galloprovincialis Lmk. (Mytilidae; Mollusca)

Genetic variation was assayed electrophoretically at 13–16 loci in Modiolus modiolus, Mytilus edulis, and Mytilus galloprovincialis. High genetic distance ( D ) values were observed between Modiolus modiolus and Mytilus edulis (1.516 ± 0.523) and between Modiolus modiolus and Mytilus galloprovincialis (1.564 ± 0.539), whereas the distance between Mytilus edulis and M. galloprovincialis (0.167 ± 0.118) was rather low. The systematic status ot Mytilus edulis and M. galloprovincialis is discussed in relation to these lindings and the genetic distance values are used to estimate divergence times which in turn are compared with paleontological estimates. The observations of high average heterozygosity in Modiolus modiolus, and high correlations of locus heterozygosity between taxa are discussed briefly.     

Assessment of a glutathione S-transferase and related proteins in the gill and digestive gland of Mytilus edulis (L.), as potential organic pollution biomarkers

The response of the glutathione S-transferase (GST, EC 2.5.1.18) and related proteins of Mytilus edulis to environmental pollution load was assessed. Mussels were reciprocally transplanted between an industrial estuary (Douglas), a rural estuary (Youghal) and a m arine site (Bantry). In addition, m ussels were sam pled along a pollution gradient in an estuary receiving leather tannery effluent (Colligan). These latter m ussels were previously shown to be subject to oxidative stress resulting from the discharges. GST specific activity of cytosolic extracts from the gill and digestive gland tissues was determ ined for all anim als. Specific activity was shown to vary significantly in anim als from different sites, with highest specific activity always observed in sam ples (local and transplanted) taken from the industrial site. By com parison, the m ussels exposed to tannery discharges displayed no significant alteration in GST specific activity. Total intracellular glutathione (GSH) was also determ ined for sam ples taken from the Douglas and Youghal estuaries but no correlation with pollution load was observed. Using FPLC analysis, we observed no specific effect on the relative levels of the GST and the individual GST related proteins in gill or digestive gland sam ples from local or reciprocally transplanted anim als from Douglas or Youghal. The increase in GST specific activity observed in samples from the industrial estuary are indicative of a possible, specific inductive agent at this site. The results from the tannery site, by com parison, indicate that general oxidative stress does not result in elevated G ST specific activity in M. edulis.     

A comparison of the reproductive cycles of Modiolus modiolus (L.), Cerastoderma (=Cardium) edule (L.), and Mytilus edulis L. in Strangford Lough,Northern Ireland

Summary This paper examines the reproductive cycles of three ecologically important marine bivalves-Modiolus modiolus (L.), Cerastoderma edule (L.), and Mytilus edulis L. in Strangford Lough, Northern Ireland over a period of almost five years.Whilst Modiolus does not appear to become sexually mature until it is several years old, Mytilus and Cerastoderma can reproduce in their first and second years of life respectively. In Cerastoderma and Modiolus sexual maturity is preceded by a period of rapid somatic growth.The subtidal Modiolus population remained in a more or less fully ripe condition virtually throughout the period of this investigation suggesting that this particular population lacked any marked cyclical reproductive activity. We interpret this as evidence of slow but almost continuous release of gametes throughout much of the year, a suggestion which is supported by recruitment data. A small intertidal population of Modiolus in Belfast Lough monitored over a period of two years exhibited a much more seasonal cycle. Here spawning occurred mainly during the autumn and winter. These data suggest that localised environmental factors are exceedingly important in controlling the annual reproductive cycle of this species.Cerastoderma from the mid-tidal sand flats ripened rapidly during the spring and spawned over a relatively restricted period in the summer. In Strangford Lough Mytilus occurs predominantly in the low-shore and while it spawns mainly in the spring and summer the annual cycle is considerably more protracted and variable than in Cerastoderma.Variations in the duration of the spawning periods in these bivalves can perhaps be explained in terms of both environmental stability and the immediate physical conditions experienced by these particular populations. The reproductive strategies exhibited by Cerastoderma, Modiolus, and Mytilus in Strangford Lough are considered in relation to population stability and to the different patterns of mortality which characterise these species in their respective local habitats.     

The mechanics of predation by the shore crab,Carcinus maenas (L.), on the edible mussel,Mytilus edulis L.

Summary Mechanical aspects of predation by the shore crab, Carcinus maenas, on the edible mussel, Mytilus edulis, were examined. The shore crabs from the population studied utilized five distinct, largely size-related, mussel-opening techniques. Crushing the mussel umbone appeared the most successful opening method for medium-sized prey. Small mussels were crushed outright and large mussels could be opened by a slow, uneconomical, boring technique. The strengths of mussels, from an exposed shore, were tested under compression in four separate planes to determine the loads a crab would need to apply to crush the shells outright and the mechanical properties of mussels. Little inter-plane variability in compressive strength was observed, although intra-plane variability appeared high. The compressive strengths of mussels from a sheltered shore were found to be significantly higher than those from the exposed shore in the plane tested. A strain gauge was embedded in a mussel shell enabling the pattern and magnitude of forces produced by crab chelae in opening a mussel to be studied. The crab's chelae did not appear overwhelmingly strong when compared directly to the compressive strength of the crab's preferred mussel sizes. It is, therefore, postulated that crabs usually seek out and exploit weak spots in the umbone of mussels by trial and error, eventually breaking through the shell by a cumulative process of extending minute fractures in the shell substructure.     

Effects of cooling and freezing on the motility of Ostrea edulis (L., 1758) spermatozoa after thawing

The aim of this work was to evaluate the effects of temperature, cryoprotectant agents and freezing curves on sperm motility of Ostrea edulis. All phases of cryopreservation were studied (evaluation of semen motility pattern, choice of cryoprotectants and freezing rates) to restore after thawing the motility characteristics distinctive of fresh semen.To assess the temperature effects on sperm motility, semen was activated using four different temperatures (25, 18, 10 and 3 °C). Sperm aliquots were maintained inactive at these temperatures for 1 and 3 h, then activated with FSW at same temperature of conservation. Sperm was activated and incubated to 3 °C with dimethylsulfoxide (Me₂SO), ethylene glycol (EG), 1–2 propylene glycol (PG) (5%, 7%, 10% and 15% final concentrations), glycerol (GlOH; 5%, 10% and 15% final concentrations) and methanol (MetOH; 4% and 10% final concentrations) for 10, 20 and 30 min. A first evaluation of freezing rates was made by testing four freezing curves: −1, −3, −6 and −10 °C/min. Then, an optimization was made by testing four freezing curves: −2.5, −3.0, −3.5 and −4 °C/min.The selected temperature for short term conservation has been 3 °C, because only this temperature has allowed good sperm motility conservation after 3 h of dry-storage; this is a time sufficient to conduct cryopreservation procedures. The sperm showed a particular sensitivity to GlOH and PG to all tested concentrations and to 15% Me₂SO. EG and MetOH to all concentrations and Me₂SO to concentrations lower than 15% have not shown significant toxic effects. The freezing rate −3 °C/min using 15% EG has shown an highest percentage of RVF (rapid, vigorous and forward) spermatozoa (class 3, about 75% of fresh semen) and an highest sperm motility duration.     

Enzyme polymorphisms in the European oyster, Ostrea edulis L.

Extracts of the digestive diverticula of more than 300 individuals from four geographically separated populations of the European oyster, Ostrea edulis L. have been examined by electrophoresis for esterase and phosphoglucose isomerase polymorphisms. Three regions of esterase variability have been detected in the electrophoretograms. It is inferred that the variants in two of these regions are governed by two co-dominant alleles at each of two loci. Two phenotypes of phosphoglucose isomerase, which is inferred to be a dimeric enzyme encoded by two alleles at a single locus, have been observed. Allele frequencies and phenotype distributions are compared in the four populations. It is concluded that the populations differ genetically at the loci investigated, the magnitude of the differences being generally greatest between different 'physiological races'.     

Egg size and larval survival of Mercenaria mercenaria (L.) and Argopecten irradians (Lamarck)

Eggs of the hard clam, Mercenaria mercenaria (L.), and the bay scallop, Argopecten irradians (Lamarck), were graded to three size categories, split into replicates and these were replicate-sampled. A chi-square analysis compared the proportion of larvae (48-h survival) to the proportions of eggs in each initial replicate experiment. Statistically significant differences attributed to egg size were found for both species. Large eggs survived better than small eggs, while those of intermediate size showed no difference between expected and observed survival.     

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.     

Molecular genetic variation within and among isolates of QPX (Thraustochytridae), a parasite of the hard clam Mercenaria mercenaria

The thraustochytrid known as QPX (Quahog Parasite Unknown) has sporadically caused disease in the hard clam Mercenaria mercenaria along the east coast of North America since the 1960s. We hypothesized that genetically distinct QPX strains might be responsible for outbreaks of QPX disease in different areas and tested this hypothesis by comparing several QPX isolates recovered from the recent outbreak in Raritan Bay, New York with QPX strains isolated from 2 outbreaks in Massachusetts, USA. There was no variation in small subunit rDNA (SSU rDNA), 5.8S rDNA, or 4 mitochondrial gene sequences. In contrast, both of the ribosomal ribonucleic acid (rRNA) operon intergenic spacers, internal transcribed spacers 1 and 2 (ITS1 and ITS2), revealed substantial sequence variation. However, strain-specific sequences were not detected because the ITS sequence variation within QPX isolates was comparable to the variation between isolates. ITS1 sequences recovered from an infected clam by amplification with a QPX ITS2-specific primer were identical to those recovered from the QPX isolates.     

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.     

Carotenoids in Ostrea edulis L. (Bivalvia: Ostreacea) from the lagoon of Venice (Italy)

By means of columnar and thin-layer chromatography the presence of carotenoids in Ostrea edulis L. (Bivalvia: Ostreacea) from the Lagoon Venice (Italy), was studied.The following carotenoids were founds: -, -, -carotene, -carotene epoxide, lutein (free and epoxide from), zeaxanthin, diatoxanthin, mutatoxanthin and astaxanthin. The dominating carotenoids were lutein.The total contents varied within the range of 5.4–8.1 µg/g wet weight.     

Phagocytosis by hemocytes of the hard clam, Mercenaria mercenaria.

Large granular hemocytes of Mercenaria mercenaria avidly phagocytose a variety of biological particles (red blood cells of six species, yeast, and gram-positive and gram-negative bacteria) as well as polystyrene spheres. Clam hemolymph is not necessary for phagocytosis but may have some opsonic effect in certain circumstances (e.g., low temperature and low particle density). Formaldehyde treatment of red blood cells enhances susceptibility to phagocytosis. Phagocytosis by Mercenaria hemocytes in vitro appears to be a nonspecific process.     

Biodeposit production by the mussel Mytilus edulis L. on rocky shores

Seasonal changes in the amount of biodeposit (faeces and pseudofaeces) produced by the mussel Mytilus edulis L., which is one of the representative suspension-feeders in the rocky intertidal and shallow subtidal regions of Mutsu Bay, were studied in the laboratory. The effects of water temperature, light, food concentration, flow rate, body size, age, and spawning on biodeposit production were investigated. More biodeposit was produced in summer than in other seasons. Throughout the year, the amount of biodeposit was positively correlated with body size. Relatively more biodeposit was produced by smaller than by larger individuals. A M. edulis population living in one square meter was estimated to produce 9.20 kg of faeces and 2.71 kg of pseudofaeces per year (dry wt). More biodeposit was produced at water temperatures of 17.6–20.2° C than at 4.5–7.6° C and 25.2–26.0° C. The optimum temperature for biodeposit production was found to be ≈ 20.0 °C. When kept in the dark, M. edulis produced more biodeposit than in the light. When food concentration is increased, more psuedofaeces are produced; the amount of faeces, however, remains constant. With increasing flow rate, the amount o f biodeposit per h increased but the biodeposition rate decreased. Larger amounts of faeces and smaller amounts of pseudofaeces were produced by younger mussels than by older ones of a similar size. Spawning also affected biodeposit production.     

The ecology of Mytilus edulis L. (Lamellibranchiata) on exposed rocky shores

Summary Growth studies in M. edulis L. have shown that rates vary considerably according to age, size and environmental conditions. This may in part be attributed to its sessile habit, being unable to move away from the variable external conditions.The use of modal length frequency distributions is somewhat limited, since with three or more year groups represented, the growth of the majority of the population is so slow that individual year classes lose their identity. Growth boxes, containing marked animals of different ages, and set out in a variety of habitats gave information regarding local and seasonal growth rates. Disturbance rings were shown to be annual, and from them growth curves have been constructed.Growth is particularly seasonal, little or none occurring during the winter. Growth rates varied considerably with a variety of environmental factors (both biotic and physical) and some of these are discussed. Variable individual growth rates, together with slow growth of the majority of animals in mixed populations, are perhaps the major factors in producing population structures typical of this species on open shores.Survivorship curves for mussels in a variety of habitats have been constructed by following the survival of groups of marked animals. Whilst high mortalities occurred in the mid and low shore, survival in the upper shore in the absence of major predators, was greatly enhanced, resulting in established populations of considerable age. Periods of maximum mortality during spring and summer could be correlated with the abundance of major predators.The almost cosmopolitan distribution of M. edulis in the N. hemisphere has been made possible by virtue of its high reproductive capacity, successful larval dispersion and wide tolerance of environmental conditions, Its patchy and apparently erratic distribution both from one shore to annother and even on the same shore, is greatly influenced by the local and seasonal abundance of major predators. Whilst the upward extension of mussels is prevented, ultimately, by physical factors (e.g. temperature, dessication), its lower limits (and absence from the shallow sub littoral on many shores) are governed essentially by predators. The upward extension of many predators such as crabs or starfish, resulting in locally intense predation, may in turn be partly influenced by the actual topography of the shore itself.     

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.