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.     

Gametic incompatibility and genetic divergence of Pacific and Kumamoto oysters,Crassostrea gigas and C. sikamea

The biological and taxonomic separation of the Pacific oyster Crassostrea gigas (Thunberg, 1793) from the Kumamoto oyster C. sikamea (Amemiya, 1928) is affirmed by three concordant lines of evidence: (1) fixed differences in 2% of a mtDNA sequence coding for large subunit rRNA; (2) a genetic distance of 0.440 based on 19 allozyme loci, including 5 diagnostic loci (Aat-1, Idh-1, Idh-2, Mpi-1, Mdh-2); (3) one-way gametic incompatibility resulting in partial reproductive isolation in interspecific crosses. C. gigas sperm x C. sikamea egg fertilizations form viable hybrid offspring, but C. sikamea sperm do not fertilize C. gigas eggs. Divergence between these two species is mediated by differing peaks in the periods for gamete release and by one-way sperm/egg incompatibility. Two attempts to recover C. sikamea from its place of origin in southern Japan have yielded only individuals with the mitochondrial haplotype that characterizes C. gigas. We thus identify a crucial need for careful screening, management, and conservation of the cultivated populations of C. sikamea on the US west coast.     

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     

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.     

Comparative vulnerability to predators,and induced defense responses,of eastern oysters Crassostrea virginica and non-native Crassostrea ariakensis oysters in Chesapeake Bay

Management agencies are considering introducing the Suminoe oyster Crassostrea ariakensis into Chesapeake Bay, USA. It is unknown if the growth of feral populations of this non-native oyster would be regulated by the same predators that once controlled the abundance of the native eastern oyster C. virginica. In laboratory studies, we compared the relative susceptibility of juvenile diploids (shell height < 25 mm) of both oyster species to invertebrate predators of eastern oyster juveniles. Predators included four species of mud crabs [Rhithropanopeus harrisii (carapace width 7–11 mm), Eurypanopeus depressus (6–21 mm), Dyspanopeus sayi (8–20 mm), and Panopeus herbstii (9–29 mm)], the blue crab Callinectes sapidus (35–65 mm), and two sizes of polyclad flatworms (Stylochus ellipticus and possibly Euplana gracilis; planar area ≲5 mm² and ∼14 to 88 mm²). All four species of mud crab and the blue crab preyed significantly (ANOVA, P ≤ 0.05) more on C. ariakensis than on C. virginica, but predation by flatworms of both sizes did not differ significantly between oyster species. The greater susceptibility of C. ariakensis to crab predation was likely due to its shell compression strength being 64% lower than that of C. virginica (P = 0.005). To test for predator-induced enhancement of shell strength, we held oysters of both species for 54 days in the presence of, but protected from, C. sapidus and R. harrisii. Crabs were fed congeneric oysters twice weekly within each aquarium. Compared to controls, shell strength of C. virginica exposed to R. harrisii increased significantly (P < 0.043), as did shell strength of both oyster species exposed to C. sapidus (P < 0.01). Despite the changes in shell strength by both oyster species in the presence of C. sapidus, the shell of C. ariakensis remained 57% weaker than C. virginica. We conclude that, because C. ariakensis exposed to predators continued to have a weaker shell relative to C. virginica, the natural suite of crab and flatworm predators in Chesapeake Bay will likely serve to control the abundance of feral C. ariakensis. We caution that the situation in the natural environment may be sufficiently different in some locations that C. ariakensis may be able to compensate for its greater vulnerability to crab predation and hence become a nuisance species.     

Growth patterns in oysters, Crassostrea virginica, from different estuaries

We analyzed a data set collected over 15 yr, containing growth data from strains of eastern oysters, Crassostrea virginica (Gmelin, 1791), initiated from parent populations in Long Island Sound, Delaware Bay, and lower Chesapeake Bay. The long-term growth data proved to be a powerful tool for examining patterns of growth differentiation among separated populations of C. virginica. The oyster strains had been grown in a common environment in lower Delaware Bay for up to seven generations. We found that the oyster strains with origins in Long Island Sound were significantly larger over several generations than oyster strains from Delaware Bay and Chesapeake Bay. Chesapeake Bay oyster strains were larger than Delaware Bay oyster strains at 1.5 yr old, but Delaware Bay oysters were larger at 2.5 yr. Year-to-year variation in environmental conditions had a strong significant effect on absolute oyster size and the relative sizes of the oyster strains. Persistent differences between oyster strains from different origins over several generations support a hypothesis that these estuarine populations have experienced long-term genetically-based population differentiation. This result is consistent with hypotheses of population differentiation of oysters based on observations of local reproductive timing. Received: 12 August 1997 / Accepted: 26 May 1998     

Interannual, seasonal, local and body size variations in reproduction of the prawn Penaeus (Marsupenaeus) japonicus (Crustacea: Decapoda: Penaeidae) in the Ariake Sea and Tachibana Bay, Japan

 The reproductive biology of female Penaeus japonicus Bate was investigated in the Ariake Sea and Tachibana Bay (located outside the Ariake Sea), Japan from 1994 to 1996. Interannual, seasonal, individual female body size and spatial influences on the incidence of spawning were examined. The proportion of inseminated females, increased with increasing body size up to 170 mm body length (BL) and decreased thereafter. The minimum size at maturity was similar between years; 130 to 140 mm BL. The minimum size of individuals with developing ovaries or spermatophores differed from that of ripe females by 20 to 25 mm. Spawning occurred in the central part of the Ariake Sea and Tachibana Bay but rarely in the inner part. P. japonicus had a clear reproductive cycle. Spawning started earlier and ended later, occurring from mid-May to mid-October, in the eastern central part of and outside of the Ariake Sea compared with the western central part, where spawning occurred from mid-June to mid-September. Spawning dynamics differed across seasons, body sizes and areas but not across years. The seasonal peak in the proportion of ripe females varied with body size. In small individuals (130 to 169 mm BL), no peak was observed, whereas in large individuals (>170 mm BL) the proportion of ripe individuals peaked in June. The proportion of ripe individuals increased with increasing body size and was high outside the Ariake Sea. Factors causing the variation in spawning dynamics are discussed. Received: 30 November 1998 / Accepted: 12 October 1999     

Mortality of newly metamorphosed eastern oysters (Crassostrea virginica) in mesohaline Chesapeake Bay

 Stocks of eastern oysters, Crassostrea virginica (Gmelin), in mesohaline Chesapeake Bay, USA, exhibit a high degree of inter-annual and spatial variability in recruitment. We found that cumulative oyster spatfall on off-bottom collector plates, measured throughout the summer in 14 years over a span of three decades, was highly positively correlated (r ² = 0.8) with juvenile oyster recruitment on adjacent oyster bars. Total abundances of juvenile oysters on these bars were, however, generally 99.7% lower than predicted from cumulative seasonal larval settlement on collector plates. We propose that although the number of larvae metamorphosing was the key factor in determining the gross annual pattern of recruitment to these mesohaline oyster bars, the actual magnitude of recruitment was governed by post-settlement processes, such as competition for limited resources and predation. We tested the hypothesis that predation may be partly responsible for high post-settlement juvenile oyster mortality. We performed a series of 3-d field investigations over two summers (1989, 1990) at a mesohaline site, employing cages of various mesh sizes (400, 800, 1500 μm) to protect hatchery-reared spat of 0.5 to 4.0 mm shell height. Mortality rates for spat held for 3 d in the estuary (17.8%) were significantly higher (P = 0.0001) for the smallest spat (0.5 to 2.0 mm) compared with those of 2.01 to 4.0 mm (4.2%). In 1990, but not in 1989, enclosure within 400 and 800 μm mesh cages significantly (P = 0.004) increased survival during 3-d deployments (9.4 and 10.1%, respectively) compared with spat unprotected by mesh cages (21.9%). In a series of laboratory predation studies that used the entire community of invertebrates that could penetrate the cages, microscopic juvenile polyclad flatworms, Stylochus ellipticus, were the only organisms that we observed crawling into living oysters and feeding on oyster tissue. Large flatworms (50 to 200 mm²) are known to be important predators on oysters, but this ability of flatworms that were so small (<ca. 5 mm²) and translucent as to be almost invisible without magnification to feed on immediate post-metamorphic oysters has not been documented previously. Our results suggest that the rate of mortality due to predation in mesohaline Chesapeake Bay is much reduced once spat survive for 2 to 3 weeks post-metamorphosis. Thus, it is likely that predation in the 1 to 2 week period immediately after settlement may be a crucial factor in the structuring of eastern oyster populations. Received: 21 December 1998 / Accepted: 2 December 1999     

Particle sorting in bivalves: in vivo determination of the pallial organs of selection

Benthic particle feeders are exposed to a food supply varying in both quantity and quality. Previous studies have shown that bivalve molluscs deal with such fluctuating particle regimes in a variety of ways, including adjustments in pumping and ingestion rates, and selective rejection of non-nutritive particles as pseudofeces. The actual site of particle selection within the pallial cavity, however, has remained a topic of speculation. During August 1995 and January and August 1996, we exposed the oysters Crassostrea virginica (Gmelin) and C. gigas (Thunberg), and the mussel Mytilus trossulus Gould to a mixture of ground, aged Spartina alterniflora Loisel and similar-sized phytoplankton at three concentrations (10³, 10⁴, 10⁵ particles ml⁻¹). We then examined the ctenidia and labial palps by means of endoscopy and sampled, in vivo, the particulate material from various ciliated tracts, and analyzed the samples with a flow cytometer. We found that in oysters, the ctenidia are responsible for particle sorting, whereas the labial palps play an accessory role in particle selection, or function to control the volume of material to be ingested. In mussels, however, the ctenidia play little role in particle selection and simply transport particulate matter to the palps for further processing. We suggest that selection by the ctenidia of oysters is a function of their architecture (plicate, heterorhabdic). Received: 4 June 1997 / Accepted: 23 January 1998     

Mercury in a northeastern Brazilian mangrove area, a case study: potential of the mangrove oyster Crassostrea rhizophorae as bioindicator for mercury

The Channel of Santa Cruz is a mangrove area in northeastern Brasil 40 km north of Recife. Until 1991 a chlor-alkali factory discharged large amounts of mercury into the main tributary of the channel, the Rio Botafogo. The objective of this study was to assess the potential of the mangrove oyster Crassostrea rhizophorae as a bioindicator for mercury in this area and to examine the influence of condition on mercury concentrations in the oysters. The investigation was carried out in the late rainy season (July to September) 1993, in the rainy season (April to June) 1994 and in the dry season (November/December) 1994. At 11 stations in the channel, mangrove oysters, surface sediments and suspended matter were sampled, representing different compartments of the system. The distribution of mercury in the Channel of Santa Cruz exhibited for all compartments the same distinct spatial pattern with maxima in the Rio Botafogo. Seasonal variations were small and revealed no clear tendencies. In addition, mangrove oysters were transplanted from more contaminated stations to less contaminated stations and vice versa. These experiments were designed to study the oysters' capability to adjust their mercury concentrations to a changing bioavailability of mercury. The transplantation experiments suggest that the oysters are capable to adjust to changing ambient mercury availability within a few months. The transplantation experiments also revealed the importance of condition changes on the mercury concentration of the oysters. This factor should not be ignored during pollution studies. However, condition had apparently no strong influence on the spatial or seasonal mercury variations of the oysters. The mangrove oyster is a suitable bioindicator for long-term changes in mercury availability. Received: 20 August 1997 / Accepted: 15 January 1998     

Importance of cysts in the population dynamics of the red tide flagellate Heterosigma akashiwo (Raphidophyceae)

To elucidate roles of cysts in occurrences of Heterosigma akashiwo blooms, cyst dynamics were studied in northern Hiroshima Bay, the Seto Inland Sea of Japan, where H. akashiwo regularly forms red tide in June. Monthly measurements of seasonal changes in the densities of vegetative cells of H. akashiwo and their germinable cysts in surface sediments (top 1-cm layer) were made for 2 years at three stations. Vegetative cells of H. akashiwo could be detected from April through December throughout the water column, and the existence of vegetative cells was confirmed in surface waters even in winter after incubation of sampled seawater in culture medium. Germinable cysts, enumerated by the extinction dilution method, existed in sediments in all seasons, even before and after the seasonal bloom. The effects of incubation temperature on the germination of natural cysts of H. akashiwo in sediments were examined. Germination was not observed at 5 °C, was low at 10 °C, while it increased at 15 °C, and maintained a high level to 25 °C. The bottom water temperature reached 15 °C (suitable for the germination of cysts) and the surface about 18 °C or more (suitable for the growth of vegetative cells) 2 to 3 weeks before the blooms. The dark survival of H. akashiwo cysts was tested, and it was found that the cysts were viable for at least 650 d at 11 °C, and for 165 d at 25 °C, indicating a significant role of cysts in the survival during winter and summer seasons. The cysts presumably also play an important role in seeding primary populations into water columns when the bottom water reaches a suitable temperature (around 15 °C); thereafter the populations develop with great annual regularity to bloom in June. These results suggest that initiation of H. akashiwo red tides in the Seto Inland Sea could be triggered by bottom water temperature. Received: 3 July 1998 / Accepted: 12 January 1999     

Detrital flows through the feeding pathway of the oyster (Crassostrea gigas) in a tropical shallow lagoon: δ13C signals

The spatial relationships and linkage of the detrital flows among the water column, the sediment and the oyster Crassostrea gigas cultured in the water column were examined by using stable carbon isotopes (δ¹³C) in a tropical shallow lagoon from October 1996 to June 1997. The lagoon is located in southwestern Taiwan and is isolated from the sea by sand barriers except at two tidal inlets. It receives freshwater mainly from two rivers. A total of 12 stations were set up along three transect lines, each running across the lagoon from riverine to tidal inlet localities. The δ¹³C values of the water-column POM exhibited a marked sea–river gradient, with values depleted from a high of −21.7‰ at seaward stations to a low of −28.2‰ at riverine stations; those in the sedimentary POM (<62 μm grain size) also revealed this trend, but to a lesser extent. Oysters of two known ages, 6 months old (“old oysters”) and newly settled individuals (“young oysters”), were transplanted from one station to each of the remaining stations, while some were left at the original station. Values of δ¹³C in the muscle of transplanted oysters changed in parallel with the sea–river gradient of δ¹³C in POM (decreasing from −16.0 to −18.5‰ in old oysters and from −16.8 to −21.9‰ in young ones). The spatial sea–river gradient of the oyster's δ¹³C is related not only to the distance between the site that the oyster inhabits and sea or riverine environment, but also to the tidal flow pattern that surrounds its feeding place. Although the δ¹³C value of the sedimentary POM was correlated with that of the water-column POM, the δ¹³C value of the oyster tissue was significantly correlated with that of the water-column POM, but not with that of the sedimentary POM. This suggests that the oyster feeds primarily on water-column rather than sedimentary POM. Received: 30 April 1999 / Accepted: 15 December 1999     

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.     

Population genetics of Crassostrea ariakensis in Asia inferred from microsatellite markers

Crassostrea ariakensis is an important aquacultured oyster species in Asia, its native region. During the past decade, consideration was given to introducing C. ariakensis into Chesapeake Bay, in the United States, to help revive the declining native oyster industry and bolster the local ecosystem. Little is known about the ecology and biology of this species in Asia due to confusion with nomenclature and difficulty in accurately identifying the species of wild populations in their natural environment. Even less research has been done on the population genetics of native populations of C. ariakensis in Asia. We examined the magnitude and pattern of genetic differentiation among 10 wild populations of C. ariakensis from its confirmed distribution range using eight polymorphic microsatellite markers. Results showed a small but significant global θ _ST (0.018), indicating genetic heterogeneity among populations. Eight genetically distinct populations were further distinguished based on population pairwise θ _ST comparisons, including one in Japan, four in China, and three populations along the coast of South Korea. A significant positive association was detected between genetic and geographic distances among populations, suggesting a genetic pattern of isolation by distance. This research represents a novel observation on wild genetic population structuring in a coastal bivalve species along the coast of the northwest Pacific.     

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.     

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.     

Genetic structure of wild European populations of the invasive Pacific oyster Crassostrea gigas due to aquaculture practices

As a result of aquaculture activities, Pacific oysters Crassostrea gigas (Thunberg, 1793) have invaded European coasts. Using seven microsatellites, we found virtually no genetic differentiation between natural populations throughout the European range (from the south of the Wadden Sea (the Netherlands) to the south of France) and French cultivated oysters. The genetic homogeneity of Pacific oyster samples appears to be the result of repeated transfers from same seed stocks made for aquaculture and, to a lesser extent, widespread dispersal due to specific biological traits of this species. The only genetic differentiation of Sylt population in the north of the Wadden Sea (Germany) suggests a stronger, persistent impact of ongoing supply of new genetic material from hatchery production, corresponding to seeds selection made by breeders. All of our genetic data highlighted the importance of aquaculture practices on the genetic structure of the keystone invader C. gigas in Europe.     

Karyotype, nucleolus organiser regions and constitutive heterochromatin in Ostrea angasi (Molluscae: Bivalvia): evidence of taxonomic relationships within the Ostreidae

Chromosome preparations from gill tissue of the Australian flat oyster Ostrea angasi Sowerby were studied with conventional Giemsa staining, C-banding and silver staining techniques. A diploid complement of 2n = 20 was observed, consisting of five metacentric, three submetacentric and two subtelocentric pairs. Constitutive heterochromatin was distributed as large centromeric blocks in Chromosome Pairs 3, 6, 8, 9 and 10. The nucleolus organizer regions (NORs) were located terminally on long arms of Chromosome Pairs 9 and 10. This allowed the identification of homologous chromosomes in submetacentric and subtelocentric pairs. Intraspecific variability in NOR pattern as revealed by differences in the number of silver-stained NORs (Ag-NORs) per cell was found to be very common. Comparison of the patterns of karyotype, C-band and Ag-NORs between species of the larviparous oysters for which data have been published demonstrate that the chromosomal structure of the endemic Australian and New Zealand species O. angasi shows little similarity to the Southern Hemisphere oysters O. (Eostrea)puelchana Orbigny and Tiostrea chilensis (Philippi) and the Indo-West Pacific oyster O. denselamellosa, but very high resemblance to the European species O. edulis. Received: 12 August 1996 / Accepted: 16 September 1996     

Mitochondrial DNA analysis of the genetic relationships among populations of scad mackerel (Decapterus macarellus, D. macrosoma, and D. russelli) in South-East Asia

The genetic relationships among South-East Asian populations of the scad mackerels Decapterus macarellus, D. macrosoma and D. russelli (Pisces: Carangidae) were investigated. In 1995 and 1996, 216 fish were sampled in seven localities spanning the seas of Indonesia and were examined for restriction-site polymorphisms using ten restriction enzymes for the mitochondrial (mt) DNA control region, amplified by the polymerase chain-reaction. The inferred phylogeny of haplotypes led to the recognition of three distinct mitochondrial lineages or phylads consistent with the distinctions of current taxonomy. All 15 mtDNA haplotypes found in D. macarellus and all 9 haplotypes found in D. macrosoma were arranged as star-like clusters, suggesting recent evolutionary history. In contrast, the phylad formed by 6 haplotypes in D. russelli from the Sulawesi Sea exhibited diffuse topology, suggesting that ancestral lineages of this species have been retained to the present. Average nucleotide-divergence estimates between haplotypes of different phylads were between 0.042 and 0.135, suggesting ancient separation, in consistency with published allozyme data. High levels of haplotype diversity, but no geographical heterogeneity, was detected within D. macarellus from the Molucca Sea and the Banda Sea. Populations of D. macrosoma exhibited both significant differences between adjacent regions (Sunda Strait and Java Sea), and broadscale genetic homogeneity from the South China Sea to the Sulawesi Sea via the Java Sea and Makassar Strait. The geographic isolation of the D. macrosoma population sampled in the Sunda Strait suggests that this region constitute a sharp transition zone between the Indian Ocean and the Sunda Shelf. Near-monomorphism of haplotypes and low nucleotide diversity (d _X) were observed in the samples of D. macrosoma from the continental shelf (haplotype-diversity estimates, h, = 0.00 to 0.25 ± 0.08 and d _X = 0.000 to 0.002). This was in contrast to the comparatively high haplotype and nucleotide diversities observed in other pelagic fish species including D. macarellus (h = 0.82 ± 0.05, d _X = 0.012 to 0.015) and D. russelli (h = 0.63 ± 0.12, d _X = 0.016), and in the oceanic D. macrosoma population sampled in the Sunda Strait (h = 0.67 ± 0.31, d _X = 0.005). We hypothesise that this may be the consequence of recent and perhaps repeated bottleneck events that have affected the D. macrosoma population sampled on the continental shelf. Received: 29 September 1997 / Accepted: 3 September 1999