Crassostrea Ariakensis Research Articles

Ultrastructural comparison of Bonamia spp. (Haplosporidia) infecting ostreid oysters.

The ultrastructure of Bonamia from Ostrea angasi from Australia, Crassostrea ariakensis from the USA, O. puelchana from Argentina and O. edulis from Spain was compared with described Bonamia spp. All appear conspecific with B. exitiosa. The Bonamia sp. from Chile had similarities to the type B. exitiosa from New Zealand (NZ), but less so than the other forms recognized as B. exitiosa. Two groups of ultrastructural features were identified; those associated with metabolism (mitochondrial profiles, lipid droplets and endoplasmic reticulum), and those associated with haplosporogenesis (Golgi, indentations in the nuclear surface, the putative trans-Golgi network, perinuclear granular material and haplosporosome-like bodies). Metabolic features were regarded as having little taxonomic value, and as the process of haplosporogenesis is not understood, only haplosporosome shape and size may be of taxonomic value. However, the uni-nucleate stages of spore-forming haplosporidians are poorly known and may be confused with Bonamia spp. uni-nucleate stages. The many forms of NZ B. exitiosa have not been observed in other hosts, which may indicate that it has a plastic life cycle. Although there are similarities between NZ B. exitiosa and Chilean Bonamia in the development of a larger uni-nucleate stage and the occurrence of cylindrical confronting cisternae, the clarification of the identity of Chilean Bonamia must await molecular studies.

Bonamia exitiosa transmission among, and incidence in, Asian oyster Crassostrea ariakensis under warm euhaline conditions.

Previously reported in Australia, New Zealand, and more recently in Europe, the protistan parasite Bonamia exitiosa was also reported in the mid-Atlantic region of the USA after causing serious mortalities there in the Asian oyster Crassostrea ariakensis. At the time, this oyster was being considered for introduction, and the potential consequences of introducing this species were being assessed using field and laboratory studies. B. exitiosa emerged as the most serious disease threat for this oyster species, especially under warm euhaline conditions and for oysters <50 mm in size. To better evaluate how quickly this parasite may be able to spread among C. ariakensis, we investigated B. exitiosa transmission and incidence in C. ariakensis. During a first trial, potential direct transmission of B. exitiosa was evaluated by cohabitating infected C. ariakensis with uninfected C. ariakensis under in vivo quarantine conditions. In a second experiment, B. exitiosa incidence was estimated in situ by determining its prevalence in C. ariakensis deployed in an enzootic area after 4, 7, 14, 21 and 28 d of exposure. Results suggest that under warm euhaline conditions B. exitiosa can be transmitted among C. ariakensis without requiring any other parasite source and that parasite incidence may be at least as high as 40% after only 4 d exposure to an enzootic area. These results underscored the severity of the bonamiasis disease threat to C. ariakensis and provided further evidence that efforts to build an aquaculture industry based on C. ariakensis in the eastern USA might have been thwarted by parasitic disease.

Mitochondrial DNA sequence analysis from multiple gene fragments reveals genetic heterogeneity of Crassostrea ariakensis in East Asia

The native Asian oyster, Crassostrea ariakensis is one of the most common and important Crassostrea species that occur naturally along the coast of East Asia. Molecular species diagnosis is a prerequisite for population genetic analysis of wild oyster populations because oyster species cannot be discriminated reliably using external morphological characters alone due to character ambiguity. To date there have been few phylogeographic studies of natural edible oyster populations in East Asia, in particular this is true of the common species in Korea C. ariakensis. We therefore assessed the levels and patterns of molecular genetic variation in East Asian wild populations of C. ariakensis from Korea, Japan, and China using DNA sequence analysis of five concatenated mtDNA regions namely; 16S rRNA, cytochrome oxidase I, cytochrome oxidase II, cytochrome oxidase III, and cytochrome b. Two divergent C. ariakensis clades were identified between southern China and remaining sites from the northern region. In addition, hierarchical AMOVA and pairwise Φ ST analyses showed that genetic diversity was discontinuous among wild populations of C. ariakensis in East Asia. Biogeographical and historical sea level changes are discussed as potential factors that may have influenced the genetic heterogeneity of wild C. ariakensis stocks across this region.

Allograft inflammatory factor 1 functions as a pro-inflammatory cytokine in the oyster, Crassostrea ariakensis.

The oyster Crassostrea ariakensis is an economically important bivalve species in China, unfortunately it has suffered severe mortalities in recent years caused by rickettsia-like organism (RLO) infection. Prevention and control of this disease is a priority for the development of oyster aquaculture. Allograft inflammatory factor-1 (AIF-1) was identified as a modulator of the immune response during macrophage activation and a key gene in host immune defense reaction and inflammatory response. Therefore we investigated the functions of C. ariakensis AIF-1 (Ca-AIF1) and its antibody (anti-CaAIF1) in oyster RLO/LPS-induced disease and inflammation. Ca-AIF1 encodes a 149 amino acid protein containing two typical Ca2+ binding EF-hand motifs and shares a 48–95% amino acid sequence identity with other animal AIF-1s. Tissue-specific expression analysis indicates that Ca-AIF1 is highly expressed in hemocytes. Significant and continuous up-regulation of Ca-AIF1 is detected when hemocytes are stimulated with RLO/LPS (RLO or LPS). Treatment with recombinant Ca-AIF1 protein significantly up-regulates the expression levels of LITAF, MyD88 and TGFβ. When anti-CaAIF1 antibody is added to RLO/LPS-challenged hemocyte monolayers, a significant reduction of RLO/LPS-induced LITAF is observed at 1.5–12 h after treatment, suggesting that interference with Ca-AIF1 can suppress the inflammatory response. Furthermore, flow cytometric analysis indicated that anti-CaAIF1 administration reduces RLO/LPS-induced apoptosis and necrosis rates of hemocytes. Collectively these findings suggest that Ca-AIF1 functions as a pro-inflammatory cytokine in the oyster immune response and is a potential target for controlling RLO infection and LPS-induced inflammation.

Hybridization between Crassostrea hongkongensis and Crassostrea ariakensis at Different Salinities

AbstractTo evaluate the survival and growth performance of hybrid larvae and juveniles, we conducted hybridization experiments between Crassostrea hongkongensis and Crassostrea ariakensis at three salinities (15, 23, and 30 ppt). Hybrid fertilization occurred in one direction (C. hongkongensis ♀ and C. ariakensis ♂), and the fertilization rates did not differ significantly among the three trial salinities. Molecular genetics analysis revealed that hybrid progenies were true hybrids. Hybrid juveniles survived better than C. hongkongensis juveniles at a salinity of 30 ppt. Hybrid larvae and juveniles showed intermediate growth between C. hongkongensis and C. ariakensis progenies. The growth of hybrid juveniles was significantly greater than that of C. hongkongensis juveniles at a salinity of 30 ppt. Overall, C. hongkongensis has a high economic value but favorable hatchery and farm of this species is restricted to lower salinity sites. Because the hybrids were shown to have greater growth and survival characteristics at high salinity, hybrids may be a more desirable candidate to be cultured in those areas of China with higher salinities, such as the Bohai Sea of China where the salinity is usually 30 ppt.

Effect of simultaneous variation in temperature and ammonia concentration on percent fertilization and hatching in Crassostrea ariakensis

The combined effects of temperature and ammonia concentration on the percent fertilization and percent hatching in Crassostrea ariakensis were examined under laboratory conditions using the central composite design and response surface methodology. The results indicated: (1) The linear effects of temperature and ammonia concentration on the percent fertilization were significant (P<0.05), and the quadratic effects were highly significant (P<0.01). The interactive effect between temperature and ammonia concentration on the percent fertilization was not significant (P>0.05). (2) The linear effect of temperature on the percent hatching was highly significant (P<0.01), and that of ammonia concentration was nonsignificant (P>0.05). The quadratic effects of temperature and ammonia concentration on the percent hatching were highly significant (P<0.01). The interaction on the percent hatching was not significant (P>0.05). Temperature was more important than ammonia in influencing the fertilization and hatching in C. ariakensis. (3) The model equations of the percent fertilization and hatching towards temperature and ammonia concentration were established, with the coefficients of determination R2=99.4% and 99.76%, respectively. Through the lack-of-fit test, these models were of great adequacy. The predictive coefficients of determination for the two model equations were as high as 94.6% and 98.03%, respectively, showing that they could be used for practical projection. (4) Via the statistical simultaneous optimization technique, the optimal factor level combination, i.e., 25°C/0.038mgmL−1, was derived, at which the greatest percent fertilization 95.25% and hatching 83.26% was achieved, with the desirability being 97.81%. Our results may provide advantageous guidelines for the successful reproduction of C. ariakensis.

Shell Hardness and Compressive Strength of the Eastern Oyster, Crassostrea virginica, and the Asian Oyster, Crassostrea ariakensis

The valves of oysters act as a physical barrier between tissues and the external environment, thereby protecting the oyster from environmental stress and predation. To better understand differences in shell properties and predation susceptibilities of two physiologically and morphologically similar oysters, Crassostrea virginica and Crassostrea ariakensis, we quantified and compared two mechanical properties of shells: hardness (resistance to irreversible deformation; GPa) and compressive strength (force necessary to produce a crack; N). We found no differences in the hardness values between foliated layers (innermost and outermost foliated layers), age class (C. virginica: 1, 4, 6, 9 years; C. ariakensis: 4, 6 years), or species. This suggests that the foliated layers have similar properties and are likely composed of the same material. The compressive force required to break wet and dry shells was also not different. However, the shells of both six- and nine-year-old C. virginica withstood higher compressive force than C. virginica shells aged either one or four, and the shells of C. ariakensis at both ages studied (4- and 6-years-old). Differences in ability to withstand compressive force are likely explained by differences in thickness and density between age classes and species. Further, we compared the compressive strength of differing ages of these two species to the crushing force of common oyster predators in the Chesapeake Bay. By studying the physical properties of shells, this work may contribute to a better understanding of the mechanical defenses of oysters as well as of their predation vulnerabilities.

Correction: HMGB in Mollusk Crassostrea ariakensis Gould: Structure, Pro-Inflammatory Cytokine Function Characterization and Anti-Infection Role of Its Antibody

The times listed in the second paragraph are incorrect in the sentence: Administration of anti-CaHMGB significantly reduced the expression levels of LITAF to 206.06, 28.73 and 29.14 times higher compared to the control group at 4 h, 8 h and 12 h post-challenge, respectively, with reduction rates around 37.71%, 89.99% and 86.62%, respectively. The correct times should be: 3.88 and 3.90 at 8 h and 12 h post-challenge. And the reduction rate at 8 h post-challenge should be 86.49% rather than 89.99%.

Cloning and expression analysis of P-glycoprotein gene in Crassostrea ariakensis

P-glycoprotein (P-gp), as an ATP-binding cassette transporter, can transport a variety of substrates across the cell membrane in human and animals, even in aquatic animals. Multixenobiotic resistance phenomenon (MXR) related to the P-gp has been identified and characterized in several aquatic organisms, however, the nature of this transporter and the structural basis for the resistance are still poorly understood. In this study, to provide an insight into the roles of P-gp in resistance of bivalve against environmental pollution and toxins, complete cDNA of P-gp gene in Crassostrea ariakensis was cloned and analyzed. The accumulation of okadaic acid (OA), a main component of diarrhetic shellfish poisoning (DSP) toxins, and expression of P-gp in gills, digestive glands and mantle of C. ariakensis, were detected upon exposure to Prorocentrum lima, a known producer of DSP toxins. The complete sequence of oyster P-gp showed a high identity to MDR/P-gp/ABCB proteins from other species and contained some classical features of ABCB transport proteins. Upon exposure to P. lima for 72h, the OA level increased 2.5-fold in gills and 6.4-fold in digestive glands of C. ariakensis. Meanwhile, the expression of P-gp mRNA increased 2.4-fold in gills and decreased slightly in digestive glands. No significant difference in P-gp protein expression was found by western blot in all of the tissues analyzed in P. lima-exposed oysters. Taking all of our results into account, we speculate that P-gp likely plays a role in the response to DSP toxins in C. ariakensis.

Detection and characterization of Bonamia ostreae in Ostrea edulis imported to China

The protozoan parasite Bonamia ostreae is a destructive pathogen of flat oysters and has been reported to be widespread in Europe and North America. The biological characteristics of this unicellular parasite are still not fully understood. In this study, 104 Ostrea edulis imported from the USA to the Guangdong province of China for consumption were examined for Bonamia infection. PCR assay, combined with restriction fragment length polymorphism, sequencing and BLAST analysis, showed that B. ostreae DNA could be detected in 1 of the 104 oyster samples. Light microscopy revealed Bonamia-like organisms in the oyster. PCR assay and fluorescent in situ hybridization showed that B. ostreae organisms were present and retained their integrity after 4 wk in culture. Acridine orange-ethidium bromide staining indicated that the B. ostreae were still alive. In conclusion, B. ostreae was present in oysters imported to China. More importantly, the parasite was able to survive for at least 4 wk of in vitro culture at 4°C, which further implied a long-term transmission risk of B. ostreae. Considering the wide culture beds of Crassostrea ariakensis and C. gigas in China, and that C. ariakensis and C. gigas are susceptible hosts or reservoirs of B. ostreae, our study highlights the potential risk of introducing B. ostreae by importing O. edulis from a Bonamia endemic area.

Molecular and histological identification of Marteilioides infection in Suminoe Oyster Crassostrea ariakensis, Manila Clam Ruditapes philippinarum and Pacific Oyster Crassostrea gigas on the south coast of Korea

The oyster ovarian parasite Marteilioides chungmuensis has been reported from Korea and Japan, damaging the oyster industries. Recently, Marteilioides-like organisms have been identified in other commercially important marine bivalves. In this study, we surveyed Marteilioides infection in the Manila clam Ruditapes philippinarum, Suminoe oyster Crassostrea ariakensis, and Pacific oyster Crassostrea gigas, using histology and Marteilioides-specific small subunit (SSU) rDNA PCR. The SSU rDNA sequence of M. chungmuensis (1716bp) isolated from C. gigas in Tongyoung bay was 99.9% similar to that of M. chungmuensis reported in Japan. Inclusions of multi-nucleated bodies in the oocytes, typical of Marteilioides infection, were identified for the first time in Suminoe oysters. The SSU rDNA sequence of a Marteilioides-like organism isolated from Suminoe oysters was 99.9% similar to that of M. chungmuensis. Marteilioides sp. was also observed from 7 Manila clams of 1840 individuals examined, and the DNA sequences of which were 98.2% similar to the known sequence of M. chungmuensis. Unlike Marteilioides infection of Pacific oysters, no remarkable pathological symptoms, such as large multiple lumps on the mantle, were observed in infected Suminoe oysters or Manila clams. Distribution of the infected Manila clams, Suminoe oysters and Pacific oysters was limited to small bays on the south coast, suggesting that the southern coast is the enzootic area of Marteilioides infection.

Habitat values for artificial oyster (Crassostrea ariakensis) reefs compared with natural shallow-water habitats in Changjiang River estuary

Oyster reefs have an equivalent, complex 3-dimensional structure to vegetated habitats and may provide similar functions in estuarine environments. Nevertheless, few studies have compared oyster reefs with adjacent natural shallow-water habitats. Here the resident benthic macroinvertebrate communities in an artificial oyster (Crassostrea ariakensis) reef and in adjacent natural estuarine shallow-water habitats (salt marsh, intertidal mudflat, and subtidal soft bottom) in the Changjiang (Yangtze) River estuary were described. The mean total densities and biomass, Margalef’s species richness, Pielou’s evenness and Shannon-Weaver biodiversity indices of the resident benthic macroinvertebrate communities differed significantly among the habitats. Significantly higher densities and biomass of benthic macroinvertebrates occurred in the oyster reef compared with the other three habitats. Ordination plots showed a clear separation in benthic macroinvertebrate communities among the four habitat types. The results demonstrated that the artificial oyster reef supported distinct and unique benthic communities, playing an important role in the complex estuarine habitat by supplying prey resources and contributing to biodiversity. In addition, the results suggested that the oyster reef had been restored successfully.

Fertilization, Survival, and Growth ofCrassostrea hongkongensis♀ ×Crassostrea ariakensis♂ Hybrids in Northern China

Hybrids between Crassostrea hongkongensis and Crassostrea ariakensis were produced to evaluate their aquaculture potential in northern China. Fertilization success was asymmetrical; C. hongkongensis eggs can be fertilized by C. ariakensis sperm, but not vice versa. Molecular genetics analysis revealed that hybrid progeny were true hybrids. Larval growth of hybrids was intermediate between C. hongkongensis and C. ariakensis, whereas larval survival in hybrid families ranged from 0.3–37%. The progeny of C. hongkongensis did not survive during an outdoor overwinter phase, whereas hybrids and C. ariakensis showed similar survival rates (>72%). Mean shell height of hybrid adults at day 360 was 54% larger than that of C. hongkongensis adults, and was 25% larger than that of C. ariakensis adults. Morphology of the hybrid adult was similar to that of C. hongkongensis. On the basis of high overwinter survival and faster growth than either parental species, we suggest that the hybrid between C. hongkongensis and C. ariakensis may substitute for C. hongkongensis as a cultured species in northern China.

Survival, Acid—Base Balance, and Gaping Responses of the Asian OysterCrassostrea ariakensisand the Eastern OysterCrassostrea virginicaDuring Clamped Emersion and Hypoxic Immersion

The eastern oyster Crassostrea virginica has a remarkable ability to withstand low oxygen conditions; however, many of the biochemical mechanisms by which that tolerance is accomplished remain poorly understood. In addition, little is known about hypoxia tolerances and adaptations of the Asian oyster Crassostrea ariakensis. By comparing these closely related species, we may learn more about the physiological mechanisms responsible for the hypoxia tolerance of C. virginica. We assessed the time to mortality and gaping responses in both species during hypoxia. Adult C. ariakensis died earlier than C. virginica and gaped more often and wider than C. virginica. Gaping by either species was associated with acidification of the ambient seawater. We also compared the hemolymph pH of emersed and clamped C. virginica and C. ariakensis between 0 h and 24 h. In both species, hemolymph pH declined over time, and C. ariakensis hemolymph became significantly more acidic throughout the study period than that of C. virginica. The infection levels of Perkinsus marinus observed in our samples were not correlated with hemolymph pH changes in either species. Our findings demonstrate fundamental differences in survival and suggest important physiological differences exist between these species.

Application of an Ecological Risk Assessment for Evaluation of Alternatives Considered for Restoration of Oysters in Chesapeake Bay: Background and Approach

ABSTRACT Oyster populations in Chesapeake Bay, USA, declined precipitously over the past three decades, and on-going efforts to restore the native oysters to former abundance were considered to be ineffective. Maryland and Virginia natural resource agencies proposed the introduction of a non-native Asian oyster (Crassostrea ariakensis) that is resistant to diseases affecting the native oyster and well adapted to the Chesapeake Bay environment. Numerous stakeholders raised concerns about potential adverse consequences of an introduction of a non-native species into a new environment. In response, state and federal agencies determined that an Environmental Impact Statement (EIS) should be prepared to address the environmental consequences of such an introduction as well as of seven other oyster restoration alternatives, including several involving only the native oyster. Preparation of an Ecological Risk Assessment (ERA) of the proposed action as well as all alternatives was an integral element of EIS preparation. This series of articles describes several different analyses that contributed to and collectively comprised the ERA conducted as input to the EIS. The final article of this series in HERA describes how the ERA and EIS findings were taken into account in the final decision on the preferred restoration alternative by state and federal agencies.

Identification and function study of a novel AIF1 from the oyster, Crassostrea ariakensis Gould

Identification and function study of a novel AIF1 from the oyster, Crassostrea ariakensis Gould

MAPK-activated CA-TSP, hemocyanin and lectin-like proteins form a protein complex in the hemocyte of oyster Crassostrea ariakensis Gould

MAPK-activated CA-TSP, hemocyanin and lectin-like proteins form a protein complex in the hemocyte of oyster Crassostrea ariakensis Gould

Evaluating the Risk of Establishing a Self-Sustaining Population of Non-Native Oysters Through Large-Scale Aquaculture in Chesapeake Bay

ABSTRACT A multistep invasive species methodology was used to evaluate the probability that the deployment of sterile non-native oysters (Crassostrea ariakensis, called Asian oyster) in field-based aquaculture could give rise to a reproductive population in Chesapeake Bay. Several pathways that could potentially lead to the unintentional release of reproductive C. ariakensis from aquaculture were identified. For the four quantifiable pathways, a probability was estimated for each step based on knowledge of ecological rates. A conservative estimate for the number of reproductive C. ariakensis that could arise over one spawning season at a hypothetical aquaculture site was estimated. A statistical expansion of this probability estimated that it is highly likely that the cultivation of putatively sterile C. ariakensis would initiate a reproductive population in Chesapeake Bay over a 10-year time span. The potential benefits of restoration actions involving non-native species in estuarine ecosystems must be weighed against potential ecological risks. A probabilistic approach can provide a useful method for summarizing the risk of an unintended introduction that may occur despite a reasonable and presumably “safe” approach for realizing benefits of a non-native species in aquaculture. The results presented here have important implications for future proposals involving the commercial use of non-native species in natural ecosystems.

Role of Ecological Risk Assessment Findings in Agency Decision-Making Regarding Oyster Restoration in Chesapeake Bay

ABSTRACT Failure of on-going management programs to restore oyster populations in Chesapeake Bay, USA, prompted state and federal agencies to consider the introduction of the non-native Asian oyster (Crassostrea ariakensis). An ecological risk assessment (ERA) of the proposed introduction was an essential element in preparation of a programmatic environmental impact statement (PEIS). The ERA had to assess risks of not only the proposed action (Asian oyster introduction) but also of the eight alternatives evaluated in the PEIS. The ERA suggested that the risk that the Asian oyster would not provide ecosystem services similar to those afforded by the native Eastern oyster was low, but there was moderate uncertainty associated with that conclusion. There was a non-zero risk of self-sustaining Asian oyster populations becoming established even if aquaculture with triploid, purportedly sterile Asian oysters were to be permitted. Nearly all of the risk conclusions had associated moderate to high uncertainty, not providing the level of proof that the agencies felt sufficient to justify proceeding with any action involving the Asian oyster. The irreversible nature of an introduction of the species bolstered that decision. Maryland and Virginia agencies have implemented numerous actions focused on the native oyster, but the outcome of these on-going actions is not yet known.

Development of an Immunological Probe to Quantify Reproductive Effort in the Suminoe Oyster,Crassostrea ariakensis(Gould 1861)

A polyclonal antibody specific to an egg protein of the Suminoe oyster, Crassostrea ariakensis, was developed to assess reproductive effort. After 2 mo of immunization, rabbit antiserum showed strong specificity to the egg protein in an enzyme-linked immunosorbent assay (ELISA). The rabbit antioyster egg immunoglobulin G detected as little as 0.2 µg/mL of Suminoe oyster egg protein by ELISA. The quantity of eggs present in an oyster was estimated using ELISA and was expressed as a gonadosomatic index (GSI). Gonadosomatic index values of Suminoe oysters were assessed monthly from January to July 2007 at the Seomjin River estuary off the south coast of Korea. Histology indicated that most oysters were mature and ready to spawn by the middle of July. Mean GSI values for oysters collected in April, when most female oysters are in early developmental stages, varied between 0.6% and 14.0%. In July, most oysters were ready to spawn and GSI values ranged from 17.5–67.0%, with a mean of 47.7%. The potential fecundity of ripe oysters was determined by dividing the number of eggs, which was estimated by ELISA, by the mean dry weight of a single egg (14 ng); fecundity ranged from 162–910 million eggs. The immunological technique used in this study was affordable and sensitive enough to measure variation in the number of eggs present among gametogenic stages.