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     

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.     

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     

Mytilus trossulus in Northern Europe

From data on allozyme, nuclear DNA and mitochondrial DNA markers, we show that the originally North Pacific/Northwest Atlantic mussel Mytilus trossulus is widespread on North European coasts, earliM. trossuluser thought to be inhabited only by Mytilus edulis. Several local occurrences of , interspersed with a dominant M. edulis, were recorded on the North Sea, Norwegian Sea and Barents Sea coasts of Norway and the Barents and White Sea coasts of Kola Peninsula in Russia. The proportion of M. trossulus genetic background observed at any one site varied from 0 to 95%. These new occurrences are not related to the previously known, introgressed M. trossulus population that occupies the Baltic Sea. The new northern occurrences retain both the F and M M. trossulus mitochondria, which have been lost from the Baltic stock. While hybridization takes place wherever M. trossulus and M. edulis meet, the extent of hybrization varies between the different contact areas. Hybrids are rare, and the hybrid zones are bimodal in the northern areas; more interbreeding has taken place further south in Norway, but even there genotypic disequilibria are higher than those in the steep transition zone between the Baltic mussel and M. edulis: there is no evidence of a collapse toward a hybrid swarm unlike in the Baltic. The Barents and White Sea M. trossulus are genetically slightly closer to the NW Atlantic than NE Pacific populations, while the Baltic mussel has unique features distinguishing it from the others. We postulate that the presence of M. trossulus in Northern Europe is a result of repeated independent inter- or transoceanic cryptic invasions of various ages, up to recent times.     

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.     

Elevated pCO2 causes developmental delay in early larval Pacific oysters, Crassostrea gigas

Increasing atmospheric CO₂ equilibrates with surface seawater, elevating the concentration of aqueous hydrogen ions. This process, ocean acidification, is a future and contemporary concern for aquatic organisms, causing failures in Pacific oyster (Crassostrea gigas) aquaculture. This experiment determines the effect of elevated pCO₂ on the early development of C. gigas larvae from a wild Pacific Northwest population. Adults were collected from Friday Harbor, Washington, USA (48°31.7′N, 12°1.1′W) and spawned in July 2011. Larvae were exposed to Ambient (400 μatm CO₂), MidCO₂ (700 μatm), or HighCO₂ (1,000 μatm). After 24 h, a greater proportion of larvae in the HighCO₂ treatment were calcified as compared to Ambient. This unexpected observation is attributed to increased metabolic rate coupled with sufficient energy resources. Oyster larvae raised at HighCO₂ showed evidence of a developmental delay by 3 days post-fertilization, which resulted in smaller larvae that were less calcified.     

Depuration of twelve trace metals in tissues of the oysters Crassostrea gigas and C. virginica

The depuration of 12 trace metals in the mantle, gill, digestive gland, and kidney of Crassostrea gigas and C. virginica was investigated under natural field conditions; oysters from a relatively contaminated environment (Redwood Creek in south San Francisco Bay) were transplanted to a relatively clean environment (Tomales Bay). In the transplanted oysters, the digestive gland and kidney depurated Cd, Cu, Hg, Ag, and Zn more readily than the mantle and gill. Other trace metals As, Fe, Mn, Ni and Se showed varying depuration patterns. The results for Cr and Pb were inconclusive, since initial concentrations were too low to follow any losses. Interspecific differences in trace metal depuration were observed. Biological half-lives for most trace metals were on the order of 23 to 60 d for C. gigas and on the order of 70 to 180 d for C. virginica.     

Cadmium kinetics in oysters — a comparative study of Crassostrea gigas and Ostrea edulis

The accumulation of cadmium was investigated in two species of oysters [Crassostrea gigas (L.) and Ostrea edulis (L.)] from the same environment and in oysters of the same species (O. edulis) from two different environments (contaminated and uncontaminated), under controlled laboratory conditions (33‰ salinity, 10°C, 100 μg Cd l^-1) for up to 111 d in 1982. C. gigas accumulated cadmium twice as fast as O. edulis (1.07 vs 0.52 μg Cd g^-1 wet wt d^-1). Furthermore, O. edulis from an uncontaminated environment accumulated cadmium faster than O. edulis from a metal-contaminated environment (0.52 vs 0.34 μg Cd g^-1 wet wt d^-1). There was no effect of cadmium exposure on total soft-tissue copper and zinc concentrations. Investigation of cytosolic metal-binding using Sephadex G-75 gel-permeation chromatography indicated that binding to very low molecular weight ligands (MW<1000) accounted for>70% of the cytosolic zinc in all oysters and>40% of the cytosolic cadmium in all oysters except O. edulis from Conwy at 83 d. In copper-contaminated oysters, excess copper was also associated with very low molecular weight ligands. Intermediate molecular weight cadmium/copper-binding proteins (similar to metallothionein in molecular weight) were observed in the cytosol and were shown to differ between species in terms of their behavior on Sephadex G-75. Finally, the distribution of accumulated cytosolic cadmium in O. edulis from the contaminated environment was shown to have a unique distribution, i.e., there was no cadmium associated with high molecular weight cytosolic macromolecules. The data indicate that both genetic and environmental factors influence cadmium accumulation in oysters.     

Looking for the clock mechanism responsible for circatidal behavior in the oyster Crassostrea gigas

Valve activity rhythm of the oyster Crassostrea gigas is mainly driven by tides in the field, but in the laboratory, only a circadian clock mechanism has been demonstrated. In an attempt to reconcile these results, the mechanisms underlying the circatidal rhythm were studied in the laboratory under different entrainment or free-running regimes and in the field at Arcachon (44°39′N/1°09′W) in February–April 2011). Results confirm the existence of a circadian clock in C. gigas. Under entrainment regimes (12-h dark/12-h light photoperiod and tidal cycles simulated by a reversing current flow), oysters exhibited both circadian and circatidal cycles. Under free-running conditions (e.g., continuous darkness), the endogenous rhythm appeared to be circadian. There was no experimental evidence for an endogenous circatidal rhythm, even in oysters just transferred from the field, where a clear tidal cycle was expressed. There are two possible mechanisms to explain tidal behavior in C. gigas: an exogenous tidal cue that drives tidal activity and masks the circadian rhythm and an endogenous circatidal clock that is sensitive to tidal zeitgebers and runs at tidal frequency.     

Molecular population structure of the kuruma shrimp <Emphasis Type="Italic">Penaeus</Emphasis><Emphasis Type="Italic"> japonicus</Emphasis> species complex in western Pacific

In a previous study on the kuruma shrimp Penaeus japonicus from the South China Sea, we detected high genetic divergence between two morphologically similar varieties (I and II) with distinct color banding patterns on the carapace, indicating the occurrence of cryptic species. In the present study, we clarify the geographical distribution of the two varieties in the western Pacific by investigating the genetic differentiation of the shrimp from ten localities. Two Mediterranean populations are also included for comparison. Based on the mitochondrial DNA sequence data, the shrimps are separated into two distinct clades representing the two varieties. Variety I comprises populations from Japan and China (including Taiwan), while variety II consists of populations from Southeast Asia (Vietnam, Singapore and the Philippines), Australia and the Mediterranean. Population differentiation is evident in variety II, as supported by restriction profiles of two mitochondrial markers and analysis of two microsatellite loci. The Australian population is genetically diverged from the others, whereas the Southeast Asian and Mediterranean populations show a close genetic relationship. Variety I does not occur in these three localities, while a small proportion of variety II is found along the northern coast of the South China Sea and Taiwan, which constitute the sympatric zone of the two varieties. The present study reveals high genetic diversity of P. japonicus. Further studies on the genetic structure of this species complex, particularly the populations in the Indian Ocean and Mediterranean, are needed not only to understand the evolutionary history of the shrimp, but also to improve the knowledge-based fishery management and aquaculture development programs of this important biological resource.     

Divergence of mitochondrial DNA lineage of the rocky intertidal goby Chaenogobius gulosus around the Japanese Archipelago: reference to multiple Pleistocene isolation events in the Sea of Japan

Geographic variations in the mitochondrial cytochrome b gene sequence (1,062 nucleotides) of the rocky intertidal goby Chaenogobius gulosus were examined and compared against the phylogeographic pattern of its sympatric sister species Chaenogobius annularis, which we studied previously. We collected C. gulosus specimens from 17 localities around the Japanese Archipelago and Korean Peninsula. Thirty-one haplotypes were obtained. The phylogenetic tree showed three geographical lineages, of which one was distributed along the coast of the Sea of Japan, suggesting that a Pleistocene isolation event in the Sea of Japan affected the genetic divergence of this species, similar to that which was observed in C. annularis. The net nucleotide divergence between the Sea of Japan and Pacific Ocean lineages in C. gulosus was much lower than that of C. annularis. Both traditional molecular clock and hierarchical approximate Bayesian computational models suggest that different divergence events affected the genetic divergences of C. gulosus and C. annularis. Our study on the sister species pair strongly supports multiple isolation events in the Sea of Japan, leading to varying levels of genetic divergence between the Pacific Ocean and Sea of Japan lineages in the Japanese coastal marine species.     

Allozyme variation in European populations of the oyster Ostrea edulis

Variations at 22 enzyme coding loci were surveyed in 11 populations of the oyster Ostrea edulis L., which were sampled between 1988 and 1990 along the Atlantic and Mediterranean coasts of Europe. Atlantic oyster beds suffered a steady decline during the last century, and restocking of beds with oysters of foreign origin has probably resulted in a high degree of interbreeding of natural oyster stocks from all Atlantic Europe. Our study confirms the low levels of genetic variability previously reported for the oyster populations from the Atlantic coasts, and extends it to the Mediterranean coasts. The locus arginine-kinase (ARK ^*) exhibited a high degree of interpopulation differentiation (F _ST=0.289), resulting from extensive variation in gene frequencies along a geographical cline. However, the overall genetic differentiation between populations was slight, and similar to that reported for other local populations of bivalves (mean genetic distance between populations is 0.010, mean F _ST=0.062). A general pattern of increasing differentiation along the coastline in an Atlantic-mediterranean direction emerged; but genetic differentiation among the Atlantic populations was not significantly lower than that observed among the Mediterranean populations. This and other results suggest that the effects of extensive transplantation of oysters among various areas in Europe are detectable only in some particular localities. The geographical distribution of low-frequency alleles suggests a restriction to gene flow outwards from the Mediterranean Sea, across the Straits of Gibraltar.     

Time-course uptake and elimination of benzo(a)pyrene and its damage to reproduction and ensuing reproductive outputs of Pacific oyster, <Emphasis Type="Italic">Crassostrea gigas</Emphasis>

The time-course of uptake and elimination of benzo(a)pyrene (BaP) for the Pacific oyster, Crassostrea gigas and reproduction damage and reproductive outputs were studied. Sexually immature C. gigas broodstock were fed for 28 days with live algae grown in four BaP solutions of 0, 50, 500, and 5,000 μg L⁻¹ (hereafter, control, 50, 500, and 5,000 oysters) and were subsequently conditioned to maturation by a feeding with BaP-free live algae under temperature manipulation for another 28 days. The 5,000 μg L⁻¹ oysters gained a steady state concentration, around 30,000 ng g⁻¹ d.w. for digestive gland, a week earlier compared to the 500 μg L⁻¹ oysters. The earlier gain or longer persistence of the steady state concentration influenced elimination of BaP, with an eliminating trend for 500 μg L⁻¹ oysters, while no elimination for 5,000 μg L⁻¹ oysters. The maternal persistence of the steady state concentration resulted in significant damages in the reproductive success and their reproductive outputs in terms of the hatching rate and larval growth, survival, and settlement. The 50 μg L⁻¹ oysters remained far below the steady state concentration, and showed a manifest eliminating behavior during the subsequent BaP-free 28 day maturation period. The reproductive success and initial larval events of 50 μg L⁻¹ oysters were comparable to those of control. However, the damage potential of the 50 μg L⁻¹ oysters might be more significant if their maternal exposure continued beyond 28 days, since the accumulation profile at this dose was linear.     

Physiological basis of extreme growth rate differences in the spat of oyster (Crassostrea gigas)

Juvenile oysters (Crassostrea gigas) (produced in November 2009) reared under uniform hatchery conditions for 4 months were selected for extreme growth rate differences by repeatedly taking larger and smaller individuals to achieve weight differences >30× between fast (F) and slow (S) growers. The physiological basis of differential growth was analyzed in experiments in June 2010, where components of energy gain (clearance and ingestion rates and absorption efficiency), energy loss (metabolic rates) and resulting scope for growth (J h^?1) were compared for groups of F and S oysters fed three different ration levels (≈0.5, 1.5 and 3.0 mg of total particulate matter L^?1). In both F and S oysters, a higher food ration promoted asymptotic increases in energy gain rates through regulatory adjustments to clearance rates, which maintained similar absorption efficiencies across the food concentrations. No significant differences were found between growth groups in mass-specific physiological rates (i.e., per unit of body mass). However, the scaling of these rates to a common size in both groups using allometric coefficients derived for C. gigas revealed higher energy gain rates coupled with lower metabolic costs of growth in fast growers. Thus, appropriate size-standardization is essential in accounting for observed differences in growth rate. Present results are in accordance with previous reports on other bivalve species on the physiological processes underlying endogenous growth differences, suggesting that the same interpretation can be applied to the extremes of these differences.     

Mitochondrial DNA phylogeography of the harbour porpoise Phocoena phocoena in the North Pacific

Genetic structure and phylogeography of the harbour porpoise Phocoena phocoena in the North Pacific were examined using 358 bps sequences from the 5′ end of the mitochondrial DNA control region including those reported previously and newly obtained from the west Pacific. AMOVA and pairwise population φ _st estimates clearly revealed genetic differentiation between an east/south and a north/northwest group with the break along the Pacific Rim at British Columbia. In addition, nested clade phylogeographical analysis, neutrality tests, mismatch distribution analysis, genetic diversities and Mantel test, suggested that the observed genetic structure might have been influenced by contiguous range expansion with restricted gene flow in the direction from south to north along the North American coasts and east to west along the Pacific Rim in the middle to late Pleistocene.     

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.     

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.     

Are Wadden Sea tidal systems with a higher tidal range more resilient against sea level rise?

Accelerated sea level rise may have serious implications for the Wadden Sea ecosystem in its present state. If sediment accumulation rates on the extensive intertidal flats stay behind sea level rise, the flats will eventually submerge. Drowning of the flats has negative consequences for nature conservation and for coastal risk management. Based upon an evaluation of steady state relations for Wadden Sea tidal basins, Hofstede (Zeitschrift für Geomorphologie 59(3): 377-391, 2015) postulated that the capacity of these basins to balance sea level rise by accumulation on intertidal flats seems positively related to mean tidal range. In the present study, morphodynamical simulations with a numerical model were performed for two tidal basins in the German Wadden Sea to verify the empirically established hypothesis. The following conclusions are established. Larger mean tidal range improves the capacity of Wadden Sea tidal basins to balance sea level rise. Wadden Sea intertidal flats are effective sediment sinks and seem quite resilient against (higher rates of) sea level rise. Finally, subtidal gullies may constitute a significant sediment source for accumulation on intertidal flats in response to sea level rise. With respect to the limited comparability of the two investigated tidal systems, morphodynamical modelling of all Wadden Sea tidal systems should be conducted.     

Genetic structure of rhinoceros auklets,Cerorhinca monocerata,breeding in British Columbia,Alaska, and Japan

Data from eight microsatellite markers screened in 246 rhinoceros auklets (Cerorhinca monocerata) from across the North Pacific revealed multiple genetic groups. The east (North America) to west (Japan) split was clearly evident in all analyses. Within the eastern Pacific, a minimum of three genetic groups are present. Surprisingly, rhinoceros auklets from Triangle Island, British Columbia, were genetically isolated from other nearby populations, including the breeding colony on Pine Island (~100 km to the east). A fourth genetic cluster (Chowiet Is) was detected using principal coordinate’s analysis; however, sample sizes were limited. Patterns of differentiation correspond to nonbreeding distributions with the eastern and western Pacific birds spending time off the west coast of North America and Japan, respectively, and may represent historical isolation in separate refugia during the Pleistocene glaciations. The patterns of genetic structure result from a combination of historical and contemporary factors influencing dispersal of rhinoceros auklets.     

Phylogeography of the seaweed Ishige okamurae (Phaeophyceae): evidence for glacial refugia in the northwest Pacific region

Although benthic marine algae are essential components of marine coastal systems that have been influenced profoundly by past and present climate change, our knowledge of seaweed phylogeography is limited. The brown alga Ishige okamurae Yendo occurs in the northwest Pacific, where it occupies a characteristic belt in the exposed intertidal zone. To understand the patterns of genetic diversity and the evolutionary history of this species, we analyzed mitochondrial cox3 from 14 populations (221 individuals) throughout its range. The 17 haplotypes found in this study formed five distinct clades, indicating significant genetic structure. The high differentiation and number of unique (private) haplotypes may result from the recolonization of the species from glacial refugia. Three putative refugia, each with high genetic diversity, were identified: southern Korea (including Jeju Island), northern Taiwan, and central Pacific Japan. Recolonization of I. okamurae was probably determined by ocean currents and changes in sea level during the last glacial period.