Occurrence of the Kumamoto oyster Crassostrea sikamea in the Ariake Sea, Japan

The Kumamoto oyster Crassostrea sikamea is distinguished from the closely related Pacific oyster C. gigas by concordant differences in 16S rDNA, allozymes, and a one-way gametic incompatibility. After repeated failures to find this oyster in its native habitat, we speculated in 1994 that “the Kumamoto oyster may be extinct in Japan”. In September 1996, we sampled small, deep-cupped oysters from the Ariake Sea and typed these for 16S rDNA and ITS-1 DNA markers previously shown to be diagnostic for the three most common oysters in the Ariake Sea, C. gigas, C. sikamea and  C. ariakensis. Our earlier suggestion of the demise of  C. sikamea proved incorrect. Of the 256 oysters sampled, 181 (71%) were  C. gigas, 53 (21%) were C. sikamea, and 22 (9%) were  C. ariakensis; no interspecific hybrids were observed. The distributions of C. sikamea and  C. ariakensis are clumped in the Ariake Sea:  C. sikamea occurs on the eastern and northern shores,  C. ariakensis occurs only in the northern part. These results emphasize the value of molecular markers for discriminating these morphologically plastic species both in the field and in aquaculture. Received 6 July 1998 / Accepted 8 October 1998     

Population genetics of the family ostreidae. I. Intraspecific studies of Crassostrea gigas and Saccostrea commercialis

The levels of genetic variation and estimates of genetic similarity and distance were determined for populations of Crassostrea gigas (Thunberg) and Saccostrea commercialis (Iredale and Roughley). The proportion of polymorphic loci in Japanese populations of C. gigas ranged from 0.58 to 0.63, while the individual heterozygosities ranged from 0.20 to 0.22. The proportion of polymorphic loci in Australian populations of S. commercialis ranged from 0.43 to 0.46, the individual heterozygosities from 0.17 to 0.19. The genetic similarity between different geographical populations of both species was approximately 99%. The genetic distance and similarity data derived from this investigation has resulted in some provisional reclassifications. The Portuguese oyster C. angulata (Lamarck) appears to be more appropriately regarded as a recent colonized isolate of C. gigas. The New Zealand oyster S. glomerata (Gould) is considered a subspecies of the eastern Australian rock oyster S. commercialis. The Japanese Kumamoto population of C. gigas warrants reclassification as the non-sibling species C. sikamea.     

Contrasting pattern of mitochondrial population diversity between an estuarine bivalve, the Kumamoto oyster Crassostrea sikamea, and the closely related Pacific oyster C. gigas

The Kumamoto oyster (Crassostrea sikamea) shows a spatially restricted distribution, favoring estuarine tideland environment. On the other hand, the Pacific oyster (C. gigas) has a broader range of habitat. The present study compared the mitochondrial population structure between the two closely related species. For accurate species identification of oysters sampled from Japanese and East Asian continental coasts, we performed sequencing analysis of the mitochondrial DNA (mtDNA) and PCR-RFLP assay of the first internal transcribed spacer of nuclear rRNA genes. Then, we estimated the extent of population differentiation within each of C. sikamea and C. gigas based on the mtDNA data. Few haplotypes were shared among the sites of sampling in C. sikamea, which contrasted with an extensive haplotype sharing among C. gigas samples. We discuss the mechanisms of elevated population differentiation observed in C. sikamea in light of the ecology and the ancient ocean geography around the present-day habitats.     

Molecular phylogenies help resolve taxonomic confusion with Asian <Emphasis Type="Italic">Crassostrea</Emphasis> oyster species

Identification of oyster species is still largely based on phenotypic characters that are highly plastic. Prompted by the proposed introduction of the Asian oyster species Crassostrea ariakensis into the Chesapeake Bay region of the U.S.A., this study uses molecular genetic information to understand the taxonomic framework surrounding C. ariakensis and to confidently distinguish among various sympatric oyster species. Putative samples of C. ariakensis and other species of cupped oysters from across Asia were collected and phylogenetic analyses were conducted on DNA sequences of both nuclear (ITS-1) and mitochondrial (COI) regions. Trees generated based on the two independent molecular datasets were highly congruent, and indicate that many oysters collected for this study as C. ariakensis were originally misidentified. Results also indicate that C. ariakensis, C. hongkongensis and C. nippona are distinct, but closely related species. There is strong support in both analyses for a close relationship between C. gigas and C. sikamea, as well as between C. belcheri and C. gryphoides, and between C. iredalei and C. madrasensis. The parsimony analyses based on these DNA markers, however, did not provide evidence to support C. angulata as a distinct species from C. gigas. Overall, the results emphasize the need for rigorous species identification, and additional extensive and intensive sampling to more accurately determine relationships among Crassostrea species, define their geographic distributions, and establish existing sympatry patterns.     

Comparing the effect of elevated pCO2 and temperature on the fertilization and early development of two species of oysters

This study compared the synergistic effects of elevated pCO₂ and temperature on the early life history stages of two ecologically and economically important oysters: the Sydney rock oyster, Saccostrea glomerata and the Pacific oyster, Crassostrea gigas. Gametes, embryos, larvae and spat were exposed to four pCO₂ (375, 600, 750, 1,000 μatm) and four temperature (18, 22, 26, 30°C) levels. At elevated pCO₂ and suboptimal temperatures, there was a reduction in the fertilization success of gametes, a reduction in the development of embryos and size of larvae and spat and an increase in abnormal morphology of larvae. These effects varied between species and fertilization treatments with S. glomerata having greater sensitivity than C. gigas. In the absence of adaptation, C. gigas may become the more dominant species along the south-eastern coast of Australia, recruiting into estuaries currently dominated by the native S. glomerata.     

The influence of irradiance on the biochemical composition of three phytoplankton species and their nutritional value for larvae of the Pacific Oyster (Crassostrea gigas)

Three species of phytoplankton grown at high (HL) or low light (LL) were fed as saturating rations to laboratory-reared larval Crassostrea gigas. Larval C. gigas fed diets of HL grown Chaetoceros gracilis and HL grown Isochrysis aff. galbana grew faster than those fed LL grown cells of the same phytoplankton species. Faster growth of C. gigas larvae was consistently associated with increases in the percent composition of short chain saturated fatty acids (FA) 14:0+16:0 in the HL grown cells. There were no consistent and significant differences between HL and LL grown phytoplankton cells in their content of carbon, nitrogen, protein, lipid or carbohydrate. Intraspecific increases in percent composition of essential fatty acids (EFAs), 20:53 and 22:63, in the phytoplankton were not associated with improvements in the growth or survival of the oyster larvae. Oyster larvae fed diets of Phaeodactylum tricornutum with a relatively high proportion of EFAs grew more slowly than those fed C. gracilis. In this experiment the proportion of dietary EFA 20:53 was negatively correlated with oyster growth rates. The faster growing oyster larvae contained relatively more of the FAs 14:0+16:0 which may be useful as measures of larval oyster condition. After a diet of one phytoplankton species for ca. 10 d, oyster larvae acquired distinctive FA profiles resembling that of their phytoplankton prey.     

Preparation and evaluation of protein microcapsules for a marine suspension-feeder,the Pacific oyster Crassostrea gigas

Little is known of the dietary requirements of marine suspension-feeders, mainly due to difficulties in preparing and delivering microparticulate diets to animals living in an aquatic environment. The food particles must be stable in seawater and digestible by the consumer. A laboratory method for preparing cross-linked, protein-walled capsules is described in this study as a means of delivering dietary protein to marine suspension-feeders such as the oyster Crassostrea gigas. Leakage of ¹⁴C-labelled protein from capsules suspended in seawater for 24 h was reduced to less than 5% by the use of the trifunctional cross-linking agent 1,3,5-benzenetricarbonyl trichloride in wall formation. Invitro experiments indicated that capsules were susceptible to digestion by proteases and by style extracts of oysters. Oysters fed encapsulated ¹⁴C-labelled protein absorbed ¹⁴C with an efficiency of 40.2±5.0% (±1 SE). Biochemical fractionation of capsule-fed oysters indicated breakdown of ingested ¹⁴C-protein into material that was soluble in 7% trichloroacetic acid. Cross-linked, protein-walled capsules can be used successfully to deliver dietary protein to the oyster C. gigas.     

Mitochondrial cytochrome oxidase I gene sequences support an Asian origin for the Portuguese oyster Crassostrea angulata

The Portuguese oyster Crassostrea angulata (Lamarck, 1819) was long assumed to be native to the northeastern Atlantic, however, a number of lines of evidence now indicate that it is a close relative, or identical, to the Asian Pacific oyster C. gigas (Thunberg, 1793). Three hypotheses have been proposed to explain how this strikingly disjunct geographic distribution may have come about: ancient vicariance events, recent anthropogenic introduction to Asia and recent anthropogenic introduction to Europe. We have performed a molecular phylogenetic analysis of C. angulata based on mitochondrial DNA sequence data for a 579-nucleotide fragment of cytochrome oxidase I. Our results show that Portuguese oyster haplotypes cluster robustly within a clade of Asian congeners and are closely related, but not identical, to C. gigas from Japan. The mitochondrial data are the first to show that Portuguese oysters are genetically distinct from geographically representative samples of Japanese Pacific oysters. Our phylogenetic analyses are consistent with a recent introduction of C. angulata to Europe either from a non-Japanese Asian source population or from a subsequently displaced Japanese source population. Genetic characterization of Pacific oysters throughout their Asian range is necessary to fully reveal the phylogenetic relationships among Portuguese and Pacific oysters. Received: 15 April 1997 / Accepted: 12 February 1998     

Acute toxicity of mercury and selenium to Crassostrea gigas embryos and Cancer magister larvae

The possible modification of mercury toxicity by selenium in embryos of the Pacific oyster Crassostrea gigas and the larvae of the crab Cancer magister was investigated. Mercury concentration eliciting abnormal development in 50% of the oyster embryos (EC₅₀) was 5.7 g l^-1 (48 h) and mortality in 50% of the crab larvae (LC₅₀) occurred with 6.6 g l^-1 (96 h). The 48 h EC₅₀ for selenium was greater than 10,000 g l^-1 for oyster embryos and the 96 h LC₅₀ for crab zoeae was 1040 g l^-1. The response from each species, when exposed to both toxicants, revealed, that a high level of selenium (5,000 g l^-1) increased mercury toxicity. Moderate selenium concentrations (10 to 1,000 g l^-1) tended to decrease mercury toxicity, although no statistical verification could be made. The order of administration of toxicants had no effect on the response of Crassostrea gigas embryos. Early developmental stages (8 h) of C. gigas embryos were most sensitive to dissolved Hg; toxicant administration 24 h after fertilization resulted in no apparent abnormalities in development.     

Population genetics of the family ostreidae. II. Interspecific studies of the genera Crassostrea and Saccostrea

The estimated levels of genetic variation are reported for 6 Crassostrea and 3 Saccostrea species. The estimates are based on examination of 25 to 34 genetic loci. The average heterozygosities ranged from 6 to 22% for the Crassostrea species and from 17 to 19% for the Saccostrea species. A new sibling species (S. manilai) from the Philippines has been identified by means of protein electrophoresis. The genetic similarity and distance between species has been computed for each genus. Also, a pairwise comparison of loci and a dendrogram of the phylogenetic relationship of species has been constructed from the two genera. Some of the taxonomic levels during oyster speciation have been computed from the data of the electrophoretic study of the two lineages. The possibilities of oyster superspecies (e. g. C. gigas, C. virginica and S. cucullata) are discussed based on our population genetic study.     

Metal Concentration in Oyster,Crassostrea Gigas,and Sediment in Ann-Ping Mariculture Ground,Taiwan

Abstract

This study was to assess the metal contamination in oyster tissue grown in the Ann-ping mariculture ground in Taiwan. the information generated from this work also revealed general metal pollution problem for Taiwan's oyster farmers. Oysters, Crassostrea gigas, and surficial sediments collected from ten locations in Ann-ping mariculture ground in Taiwan for metals concentration (Cu, Zn, Pd, Cd, Fe and Mn) were performed. Analytical results indicated that the yearly averaged oyster copper concentrations (μg g^?1, wet weight) in oyster soft parts from Ann-ping increased from 21.3±4.1 in 1993; 24.1±6.8 in 1994; 36.8±11.9 in 1995 to 43.9±23.1 μg g^?1, wet weight, in the 1996 raising season. the mean oyster copper concentration reached a level of 50 μg g^?1, wet weight, in December 1996. This increasing trend of metal concentration in oyster tissue indicates a potential pollution source which may pose a potential disaster as green oyster incidence, which occurred on the Charting coast in 1986, in Taiwan. Sediment samples in Ann-ping mariculture ground were also collected and examined. the seasonal variation of the copper concentration in surficial sediment from Ann-ping did not show an increasing trend as observed in oyster tissue.     

Toxic dinoflagellates (Alexandrium fundyense and A. catenella) have minimal apparent effects on oyster hemocytes

The possible effect of Alexandrium spp. containing paralytic shellfish poisoning (PSP) toxins on the hemocytes of oysters was tested experimentally. In one trial, eastern oysters, Crassostrea virginica Gmelin, were exposed to bloom concentrations of the sympatric dinoflagellate, Alexandrium fundyense Balech, alone and in a mixture with a non-toxic diatom, Thalassiosira weissflogii (Grun) Fryxell et Hasle. Subsequently, another experiment exposed Pacific oysters, Crassostrea gigas Thunberg, to a mixed suspension of the sympatric, toxic species Alexandrium catenella (Whedon et Kofoid) Balech, with T. weissflogii. Measurements of numbers of oyster hemocytes, percentages of different cell types, and functions (phagocytosis, reactive oxygen species (ROS) production, and mortality) were made using flow-cytometry. During and after exposure, almost no significant effects of Alexandrium spp. upon hemocyte numbers, morphology, or functions were detected, despite observations of adductor-muscle paralysis in C. virginica and measured toxin accumulation in C. gigas. The only significant correlation found was between toxin accumulation at one temperature and higher numbers of circulating live and dead hemocytes in C. gigas. The PSP toxins are known to interfere specifically with sodium-channel function; therefore, the finding that the toxins had no effect on measured hemocyte functions suggests that sodium-channel physiology is not important in these hemocyte functions. Finally, because oysters were exposed to the living algae, not purified toxins, there was no evidence of bioactive compounds other than PSP toxins affecting hemocytes in the two species of Alexandrium studied.     

Effects of a phycotoxin,okadaic acid,on oyster heart cell survival

Okadaic acid (OA) is a dinoflagellate toxin which accumulates in shellfish producing diarrhetic shellfish poisoning (DSP) in humans. It was found that OA is a highly selective inhibitor of protein phosphatase types 1 (PP1) and 2A (PP2A) which produces a marked increase in phosphorylation of several proteins, including p38 mitogen-activated protein (MAP) kinase. The cytotoxicity attributed to OA and the effects on p38 MAP kinase and calcium current were examined in the oyster Crassostrea gigas in this study. Data showed that p38 MAP kinase is strongly expressed in oyster heart and that OA bioaccumulated in cultured heart cells. Hence the effects of OA was tested in vitro and in vivo on oysters. OA was found to (i) exert a positive chronotropic effect on cultured atrial cardiomyocytes which is related to an increase in calcium current via PKC as shown by patch clamp measurements, (ii) produce an activation/phosphorylation of MAP kinase as shown by Westernblot while the non-phosphorylated p38 remained constant during treatment, (iii) did not induce a pro-apoptotic effect. Data suggest that OA may also stimulate the anti-apoptotic pathway by phosphatase inhibition.     

Feeding current characteristics of three morphologically different bivalve suspension feeders, Crassostrea gigas, Mytilus edulis and Cerastoderma edule, in relation to food competition

Introduced Pacific oysters (Crassostrea gigas) have shown rapid expansion in the Oosterschelde estuary, while stocks of native bivalves declined slightly or remained stable. This indicates that they might have an advantage over native bivalve filter feeders. Hence, at the scale of individual bivalves, we studied whether this advantage occurs in optimizing food intake over native bivalves. We investigated feeding current characteristics, in which potential differences may ultimately lead to a differential food intake. We compared feeding currents of the invasive epibenthic non-siphonate Pacific oyster to those of two native bivalve suspension feeders: the epibenthic siphonate blue mussel Mytilus edulis and the endobenthic siphonate common cockle Cerastoderma edule. Inhalant flow fields were studied empirically using digital particle image velocimetry and particle tracking velocimetry. Exhalant jet speeds were modelled for a range of exhalant-aperture cross-sectional areas as determined in the laboratory and a range of filtration rates derived from literature. Significant differences were found in inhalant and exhalant current velocities and properties of the inhalant flow field (acceleration and distance of influence). At comparable body weight, inhalant current velocities were lower in C. gigas than in the other species. Modelled exhalant jets were higher in C. gigas, but oriented horizontally instead of vertically as in the other species. Despite these significant differences and apparent morphological differences between the three species, absolute differences in feeding current characteristics were small and are not expected to lead to significant differences in feeding efficiency.     

Combined effects of temperature and salinity on fed and starved larvae of the mediterranean mussel Mytilus galloprovincialis and the Japanese oyster Crassostrea gigas

The combined effects of temperature, salinity and nutrition on survival and growth of larvae of the Mediterranean mussel Mytilus galloprovincialis and the Japanese oyster Crassostrea gigas were studied over a period of 7 d in the laboratory. Ripe adults, collected in spring and summer 1987 from natural populations in the Bay of Arcachon, France, were induced to spawn. Larvae of both species were cultured at four temperatures (15°, 20°, 25° and 30°C), four salinities (20, 25, 30 and 35S) per temperature, and two levels of nutrition (fed and unfed) per temperature/salinity combination. The fed larvae received a mixed algal diet of 50 cells each of Isochrysis galbana and Chaetoceros calcitrans forma pumilum per microlitre. In both bivalve species, larvae survived over a wide range of temperature and salinity, with the exception of mussel larvae, which died at 30°C. Statistical analysis indicated that nutrition had the greatest effect on larval development, explaining 64 to 75% of the variance in growth of M. galloprovincialis and 54 to 70% in growth of Crassostrea gigas. Unfed mussel larvae displayed little growth. Compared with temperature, the effect of salinity was very slight. M. galloprovincialis larvae exhibited best growth at 20°C and 35S and C. gigas at 30°C and 30S.     

Fitting fertilisation kinetics models for free-spawning marine invertebrates

 To determine how fertilisation varied with sperm concentration for two species of scallop, Chlamys (Equichlamys) bifrons (Lamarck) and C. asperrima (Lamarck), we performed a simple series of sperm dilution experiments, and measured egg size and sperm swimming speeds. C. bifrons eggs were much larger (average diam=116.5 μm), and sperm swimming speeds faster (209.8 μm s⁻¹), than C. asperrima (71.2 μm, 166.0 μm s⁻¹). In both species, maximum fertilisation occurred at an ambient sperm concentration of around 100 sperm μl⁻¹; the maximum proportion of eggs fertilised was less than 0.70 in the C. bifrons experiments, but nearer 1.0 with C. asperrima. At high sperm concentrations (>100 sperm μl⁻¹), fertilisation decreased (presumably due to polyspermy) with increasing sperm concentration, but decreased more rapidly in C. bifrons than C. asperrima. A polyspermy-adjusted fertilisation kinetics model could be fitted to the experimental data, but unique parameter estimates could not be determined. Received: 7 October 1999 / Accepted: 8 July 2000     

Ultrastructural investigations on sperm penetration and gamete fusion in the ascidians Boltenia villosa and Phallusia mammillata

Sperm-egg interactions during fertilization were studied by scanning and transmission electron microscopy in the two ascidian species, Boltenia villosa (collected in 1990 near Friday Harbor, Washington, USA) and Phallusia mammillata (collected in 1990 near Brest, France). Emphasis was put on sperm-egg vitelline coat (VC) and sperm-egg plasma membrane interaction. In both species the sperm activation as revealed by translocation of the mitochondrion was exhibited by sperm binding firmly to or penetrating the VC. Vesicles are present in the sperm apex of both species. Their persistence in late stages of VC penetration suggests for them a function rather in Sperm-egg fusion than in VC penetration. In P. mammillata sperm which are about to make contact and to start fusion with the egg plasma membrane, two types of vesicles, one located at the tip and the other slightly behind the apical cap, were detected. Function and fate of these prominent vesicles remain to be determined. Membrane fusion starts between the sperm membrane of the cap-like structure and short microvilli of the egg surface and then proceeds simultaneously at several sites of the apical region. In B. villosa conspicuous microvilli which are formed at the contact site and interdigitate with the corkscrew-like sperm represent fusion sites on the egg. In both ascidian species sperm are incorporated into the egg at an acute angle. Once fully embodied the sperm lie close and almost parallel to the egg membrane. Our findings are compared with observations in Ciona intestinalis, the only other ascidian species in which Sperm-egg interactions have been thoroughly investigated so far. We suggest that observed differences in the fusion processes are based on morphological differences in sperm structure which reflect phylogenetic modifications.     

Evolution of egg size and fertilisation efficiency in sea stars: large eggs are not fertilised more readily than small eggs in the genus<Emphasis Type="Italic"> Patiriella</Emphasis> (Echinodermata: Asteroidea)

Fertilisation kinetics theory suggests that, when sperm are limiting, the larger eggs of broadcast-spawning marine organisms ought to be fertilised more frequently than smaller eggs, because they provide a bigger target for searching sperm. Whilst this effect has been demonstrated within species, it is not known if this pattern holds among species. We tested whether a large difference in egg size between congeneric seastars with contrasting planktotrophic and lecithotrophic modes of development results in differences in the likelihood of eggs being fertilised in sperm-limiting situations. Measurement of egg sizes and sperm swimming speeds led to the prediction that the sperm–egg collision rate constant for Patiriella calcar (420-µm-diameter egg) should be nine times greater than for P. regularis (140-µm-diameter egg). Although the eggs of P. calcar should be fertilised at greater rates in low sperm concentrations, they were not. When gametes were allowed to mix for 10 s, the hypothesis that P. calcar eggs required less sperm than P. regularis to ensure 50% of eggs were fertilised was rejected. When gametes were mixed for 5 min, P. regularis eggs were more frequently fertilised, but the difference was not statistically significant. We conclude there must be a difference between these species in the likelihood that when a sperm finds a conspecific egg it can successfully fertilise. This apparent uncoupling of egg size and likelihood of fertilisation suggests that fertilisation is not a major constraint on the evolution of egg size in these seastars.Communicated by G.F. Humphrey, Sydney     

Morphological and habitat divergence in the intertidal limpet Patelloida pygmaea

In the intertidal limpet Patelloida pygmaea, two distinctive morphs, the forms pygmaea and conulus, have been recorded. The former possesses a flat elongated shell, and the latter has an extremely high round shell. It has been observed in the field that pygmaea is found on oyster shells Crassostrea gigas. The form conulus uses an unusual substrate for attachment. It is found on the living shells of the intertidal gastropod Batillaria cumingi. Although conulus is normally found only on shells of Batillaria, it can also be found on oyster shells when pygmaea and Batillaria shells are not present in nature. An electrophoretic analysis of allozymes showed that these two forms are reproductively isolated from each other and coexist without gene exchange on the same mudflat. Laboratory experiments showed that pygmaea prefers oyster shells and conulus prefers Batillaria shells as substrates for attachment when both oyster and Batillaria shells are present. The form pygmaea did not attach to Batillaria shells, even when only Batillaria shells were available. However, conulus also attached to oyster shells when Batillaria shells were not available. The proportion of individuals of conulus that attached to oyster shells decreased significantly when pygmaea was attaching to the oyster shells. These results suggest that pygmaea is ecologically more specialized to living on oyster shells than conulus.     

Geographical patterns of variability at allozyme loci in the European oyster Ostrea edulis

The variability of 14 enzyme-coding genes has been analysed in samples from 19 populations of the oyster Ostrea edulis L., collected along the Atlantic and Mediterranean coasts of Europe. We found an abundance of clines, which appeared at 8 loci, including the most polymorphic (AP-2 ^*, ARK ^*, EST-4 ^*, MDH-2 ^*, ME-1 ^*, 6PGH ^*, PGI ^* and PGM ^*). Another 6 loci (ALDH ^*, EST-3 ^*, EST-5 ^*, IDH-2 ^*, MDH-1 ^*, ME-2 ^*) exhibited V-shaped patterns of gene-frequency variation, with clines at one or both sides of the Straits of Gibraltar. The observation of coincident clines at many loci can be explained by a model of secondary intergradation. The geographical location of the midpoints of the clines and V-shaped patterns suggests the existence of two ancient Atlantic and Mediterranean oyster stocks which became differentiated in allopatry and subsequently merged. Clines observed along Atlantic and/or Mediterranean coasts at the loci with V-shaped patterns must have arisen independently. The large heterogeneity observed in the levels of gene differentiation (G _ST ) across loci (G _ST ranged from 0.008 to 0.290) and important differences in estimates of gene flow obtained by different methods suggest that the populations of O. edulis are not in genetic equilibrium. Lack of population equilibrium can be due to natural selection and/or restrictions to gene flow. The average among-population variability was higher than in other oyster species that do not show incubatory habits, and represented 8.8% of the total heterozygosity. Levels of intrapopulation variability were lowest in populations from the North Atlantic, suggesting low population sizes in that area.