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The Suminoe oyster Crassostrea ariakensis (Fujita, 1913) is one of the most important ecological and fishery bivalve mollusks with a worldwide distribution. Here, we reported an improved high-quality chromosomal-level genome assembly of C. ariakensis inhabiting the South China Sea, using Nanopore technology, Illumina sequencing, and high-throughput chromosomal conformation capture analysis. The assembled genome size is 631.73 Mb, with contig N50 length of 5.36 Mb and scaffold N50 length of 61.15 Mb, and is assigned to 10 chromosomes. A total of 29,357 protein-coding genes are predicted, 96.68% of which are functionally annotated. The genome contains 347.11 Mb (54.94%) of repetitive elements and 1130 noncoding RNAs. This improved genome assembly of south C. ariakensis is an important resource for understanding oyster diversity and evolution, and provides insights into genetic improvement, protection and management of oyster resource. [ABSTRACT FROM AUTHOR]

Heat stress has received growing concerns regarding the impact on seafood quality. However, the effects of heat stress on the sensory properties of seafood remain unknown. In this study, the sensory properties of fresh oyster (Crassostrea ariakensis) treated with chronic heat stress (30 °C) for 8 weeks were characterized using electronic nose, electronic tongue, sensory evaluation, HS–SPME–GC–MS, LC–MS and transcriptomics. Overall, chronic heat stress reduced the overall sensory properties of oysters. The metabolic network constructed. based on enrichment results of 423 differential metabolites and 166 differentially expressed genes, showed that the negative effects of chronic heat stress on the sensory properties of oysters were related to oxidative stress, protein degradation, lipid oxidation, and nucleotide metabolism. The results of the study provide valuable insights into the effects of heat stress on the sensory properties of oysters, which are important for ensuring a sustainable supply of high-quality seafood and maintaining food safety. [ABSTRACT FROM AUTHOR]

Crassostrea ariakensis (Fujita, 1913) is one of the most important economic and ecological oysters that is naturally distributed along the coast of Asia, separated by the Yangtze River estuary. They are usually compared as different populations, while there is no consensus on whether C. ariakensis in northern and southern areas should be considered as two species or subspecies. Here, we analyzed morphological characteristics, COI, 16s rRNA, mitogenome sequences, and species delimitation analysis (ASAP and PTP) to resolve the intraspecific taxonomic status of the C. ariakensis. Phylogenetic and ASAP analysis highlight that C. ariakensis was divided into N-type and S-type. PTP was unable to differentiate between the two types of C. ariakensis. The divergence time of N-type and S-type C. ariakinsis is estimated to be 1.6 Mya, using the relaxed uncorrelated lognormal clock method. Additionally, significant morphological differences exist between the two groups in terms of the adductor muscle scar color. Despite these differences, the COI (0.6%) and 16S rRNA (0.6%) genetic distance differences between N-type and S-type C. ariakensis has not yet reached the interspecific level. These results suggest that N-type and S-type C. ariakensis should be treated as different subspecies and renamed as C. ariakensis ariakensis subsp. nov and C. ariakensis meridioyangtzensis subsp. nov. [ABSTRACT FROM AUTHOR]

Glycogen serves as the principal energy reserve for metabolic processes in aquatic shellfish and substantially contributes to the flavor and quality of oysters. The Jinjiang oyster (Crassostrea ariakensis) is an economically and ecologically important species in China. In the present study, RNA sequencing (RNA-seq) and assay for transposase-accessible chromatin using sequencing (ATAC-seq) were performed to investigate gene expression and chromatin accessibility variations in oysters with different glycogen contents. Analysis identified 9 483 differentially expressed genes (DEGs) and 7 215 genes with significantly differential chromatin accessibility (DCAGs) were obtained, with an overlap of 2 600 genes between them. Notably, a significant proportion of these genes were enriched in pathways related to glycogen metabolism, including “Glycogen metabolic process” and “Starch and sucrose metabolism”. In addition, genomewide association study (GWAS) identified 526 single nucleotide polymorphism (SNP) loci associated with glycogen content. These loci corresponded to 241 genes, 63 of which were categorized as both DEGs and DCAGs. This study enriches basic research data and provides insights into the molecular mechanisms underlying the regulation of glycogen metabolism in C. ariakensis. [ABSTRACT FROM AUTHOR]

Aminotransferases are enzymes found in living organisms that catalyze transfer reactions between amino acids and keto acids, crucial for amino acid metabolism and synthesis. Aminotransferase classes I and II play a vital role in regulating osmolarity, protecting cells, and improving metabolic homeostasis and cellular fitness. To investigate the characteristics of the aminotransferase class I and II gene family and their roles in osmotic pressure regulation in the Jinjiang oyster (Crassostrea ariakensis), the gene structure, chromosomal localization, and phylogeny were characterized and the genes' expression in the gill under high-salt stress was analyzed. In this study, eighteen Aminotransferase class I and II genes, including SPTLCa and SPTLCb, SPTa and SPTb, ALAT2a and ALAT2b, KAT3a and KAT3b, and ASTa, ASTb and ASTc, were identified. The physicochemical properties of 11 family members were stable, with their instability factors less than 40. Subcellular localization prediction showed that aminotransferase classes I and II were localized in the cytoplasm or mitochondria. Chromosomal localization results showed that the 18 aminotransferase class I and II genes were located on eight chromosomes. All members of this gene family had the Aminotran_1_2 structural domain which is associated with osmotic pressure regulation by adjusting the conversion reaction between amino acids and keto acids. Most gene expressions showed an initial increase followed by a decrease from 0 h to 12 h when the oysters were challenged by acute stress using artificial seawater with a salinity of 40. However, the expression of CarAGT2 and CarKAT3b genes showed an increased trend with increasing stress time. This study systematically investigated the bioinformatics characteristics of the aminotransferase class I and II gene family in C. ariakensis and their role in osmotic pressure regulation, which provides scientific data for understanding the potential functions of these genes in physiological adaptation, thereby expanding the research on osmoregulation in bivalves. [ABSTRACT FROM AUTHOR]

Jinjiang oyster Crassostrea ariakensis, a species with economic and ecological value, is distributed along the estuaries and coasts of East Asia. With the decline in natural resources, the conservation and aquaculture of this species is urgent. However, studies characterizing their shell shape remain scarce. We investigated the morphological differences in the shells of wild Jinjiang oysters from six populations (Qinzhou, Shanghai, Nantong, Qingdao, and Binzhou hard or muddy bottom) along the coast of China. The color of the shell and adductor muscle scar showed associations with temperature gradient along its geographical distribution. Oyster shape was defined by shell height to shell length ratio, and the ratio varies among geographic locations of the populations. They were found nearly round (Qinzhou and Nantong populations), oval (Qingdao and Binzhou populations), or water-droplet-shaped (Shanghai population). Binzhou populations living on muddy substrates are more elongated than those on hard substrate. In addition, we developed a method to measure the cavity volume in oysters. Correlation and path analysis showed that shell height significantly influenced cavity volume. The synergistic effect of the two factors (the height, length, and width of the shell in pairs) on the cavity volume resulted in differences between northern and southern groups: samples from the southern group (Qinzhou and Shanghai) showed correlation between shell height and shell width, while those from the northern group (Nantong, Qingdao, and Binzhou) showed correlation between shell height and shell length. All populations showed significant correlation between shell height and cavity height, and shell length and cavity length, while the correlation between shell width and cavity width was minimal, which may have been resulted from uneven shell thickness. The linear equation for shell height and cavity volume under different ratios of shell height to length was obtained. In this study, we determined that shell height has the most influence on cavity volume, and specific cavity volume fitting linear equations are given for different shell types, which may provide a reference for future oyster breeding for shell shaping. [ABSTRACT FROM AUTHOR]

Glycogen, a stored form of glucose, is an important form of energy for aquatic shellfish, contributing to the flavor and quality of the oyster. The glycogen synthase (GYS) and glycogen synthase kinase 3β (GSK3β) are two major enzymes in the glycogenesis. However, the information of the two genes in the Jinjiang oyster Crassostrea ariakensis remains limited. In this study, we identified the genes of GYS and GSK3β and further explored their function in the glycogen synthesis of C. ariakensis. The GYS and GSK3β were distributed in all tested tissues, and high expression of GYS and glycogen content were detected in the gonad, labial palp, hepatopancreas, and mantle, while the high expression of GSK3β was observed in the gill and adductor muscle. The expression of GYS was positively correlated with the glycogen content, while GSK3β was negatively correlated. Additionally, knockdown of GSK3β using RNAi decreased the GYS expression, revealing the negative regulatory effect of GSK3β on GYS. These findings enrich the research data of GSK3β and GYS involved in glycogen synthesis, providing valuable information for further research on the function of GSK3β and GYS in the glycogen synthesis process of oyster. [ABSTRACT FROM AUTHOR]

Comparing the responses of closely related species to environmental changes is an efficient method to explore adaptive divergence, for a better understanding of the adaptive evolution of marine species under rapidly changing climates. Oysters are keystone species thrive in intertidal and estuarine areas where frequent environmental disturbance occurs including fluctuant salinity. The evolutionary divergence of two sister species of sympatric estuarine oysters, Crassostrea hongkongensis and Crassostrea ariakensis, in response to euryhaline habitats on phenotypes and gene expression, and the relative contribution of species effect, environment effect, and their interaction to the divergence were explored. After a 2‐month outplanting at high‐ and low‐salinity locations in the same estuary, the high growth rate, percent survival, and high tolerance indicated by physiological parameters suggested that the fitness of C. ariakensis was higher under high‐salinity conditions and that of C. hongkongensis was higher under low‐salinity conditions. Moreover, a transcriptomic analysis showed the two species exhibited differentiated transcriptional expression in high‐ and low‐salinity habitats, largely caused by the species effect. Several of the important pathways enriched in divergent genes between species were also salinity‐responsive pathways. Specifically, the pyruvate and taurine metabolism pathway and several solute carriers may contribute to the hyperosmotic adaptation of C. ariakensis, and some solute carriers may contribute to the hypoosmotic adaptation of C. hongkongensis. Our findings provide insights into the phenotypic and molecular mechanisms underlying salinity adaptation in marine mollusks, which will facilitate the assessment of the adaptive capacity of marine species in the context of climate change and will also provide practical information for marine resource conservation and aquaculture. [ABSTRACT FROM AUTHOR]

Peptides from oysters have several bioactive functions. In this study, we identified antioxidant peptides from oysters (Crassostrea rivularis) and investigated their structure–function relationship. We used an 8 kDa molecular‐weight (MW) cut‐off membrane and semiprep reversed‐phase liquid chromatography to collect five peptides (F1–F5) and identified the highest‐abundance ion‐peak sequences AWVDY (F1), MSFRFY(F2), EPLRY(F3), RKPPWPP(F4), and YAKRCFR(F5) having MWs of 652, 850, 676, 877, and 943 Da, respectively, using ultra‐performance liquid chromatography‐quadrupole/time‐of‐flight tandem mass spectrometry. These peptides exhibited high antioxidant activities, similar to butylated hydroxytoluene, reduced glutathione, and ascorbic acid. F5 demonstrated the highest scavenging activity for DPPH radicals (IC50 = 21.75 μg/ml), hydroxyl radicals (IC50 = 18.75 μg/ml), and superoxide radicals (IC50 = 11.00 μg/ml), while F3 demonstrated the highest reducing power. Furthermore, F5 significantly protected Caco‐2 cells from H2O2‐induced oxidative damage. These results suggest that the antioxidant peptide F5 is a promising food additive that protects against oxidative damage. [ABSTRACT FROM AUTHOR]

This study explored the efficacy of multi-elements combined with chemometrics to discriminate the geographical origins of oysters (Crassostrea ariakensi). We determined 52 elements in 166 samples from four regions along the southeast coast of China. Significant regional variations of 51 elements were revealed (P < 0.05), while the principal component analysis (PCA) provided no clear regional delineations. The training models (n = 117) established on linear discriminant analysis (LDA), partial least square discriminant analysis (PLS-DA), and random forest (RF) uniformly achieved 100% predictive accuracy. The cross-validation accuracies of the final models (n = 166) derived from LDA, PLS-DA, and RF were 100%, 100%, and 99.4%, respectively. Even with the models simplified to 8 elements (Zn, Al, K, Cd, Cu, Rb, B, and Ag), high predictive and cross-validation accuracies were maintained, underscoring the robustness and algorithm flexibility of elemental profiling for accurately identifying the geographical origins of oysters. [Display omitted] • 52 elements in 166 oyster samples from four distinct regions were determined using ICP-MS. • Elemental profiling of oysters presented significant regional variations. • LDA, PLS-DA, and RF fulfilled the discriminant tasks, with high predictability and stability. • PLS-DA and RF co-recommended 8 elements for parsimony and effective models. [ABSTRACT FROM AUTHOR]

Crassostrea rivularis are the main edible economic shellfish rich in bioactive substances. Therefore, Crassostrea rivularis resources have high development prospects. This study aimed to analyze the biological activity of neutral oyster polysaccharide powder (OPS-1) to enhance the comprehensive utilization value of oysters. OPS-1 was purified from Crassostrea rivularis by hot water agglutination using DEAE-cellulose column chromatography. Experimental results showed that OPS-1 was predominantly composed of 97.41 g/100g glucose, linked by a β-glycosidic bond, and exhibited a triple-helical conformation with a molecular weight of 954.32 kDa. OPS-1 solution is relatively stable with 15.87 ± 0.66 mV of zeta potential. In addition, OPS-1 has good thermal properties and a high antioxidant capacity. The median inhibitory concentrations (IC 50 values) of OPS-1 on DPPH radicals, ABTS+ radicals, OH radicals, and Fe2+ chelating ability were 0.23, 2.96, 2.68 and 2.33 mg/mL, respectively. Meanwhile, OPS-1 could inhibit the growth of common food-borne pathogenic bacteria (Escherichia coli , Salmonella Typhimurium, and Staphylococcus aureus). The inhibition zone sizes were not significantly different from penicillin (p > 0.05). These findings indicate that OPS-1 may be a novel natural antioxidant and antimicrobial agent derived from animal sources, providing a rationale for further investigating oyster polysaccharides. [Display omitted] • OPS-1 is mainly composed of glucose linked by β-glycosidic bonds. • OPS-1 has good thermal properties and antioxidant properties. • OPS-1 is bacteriostatic against major food-borne bacteria. • OPS-1 is an excellent candidate for antimicrobial and antioxidant products. [ABSTRACT FROM AUTHOR]

The Jinjiang oyster Crassostrea ariakensis, naturally distributing in estuarine regions with low salinity, is an important economic and ecological species in China. However, studies on its genomics and population genetics remain lacking. Here, we assembled the chromosome‐level genome of a female C. ariakensis and re‐sequenced 261 individuals from five locations in China representing three typical habitats. The C. ariakensis genome was 662.9 Mb with contig N50 length of 5.9 Mb using PacBio HiFi‐CCS long reads, and 99.83% sequences were anchored onto 10 pseudochromosomes using Hi‐C data. A total of 26,354 protein‐coding genes were predicted. We identified three significantly expanded gene families which are closely associated with osmotic pressure regulation, including CDO, SLC13 and SDR. Population structure analysis revealed that the C. ariakensis from five locations were clustered into three typical groups (northern, southern and Shanghai) (K = 3) and their phylogenetic relationship was consistently correlated to their geographical distribution. Furtherly, the differentiation between northern and southern groups was clearly demonstrated by estimated population differentiation coefficient (FST = 0.1154), and the PSMC distribution showed the two groups of effective population size separated at 0.1 Ma. Meanwhile gene flow from southern to Shanghai was detected. Selective sweep analysis between northern and southern group detected genes associated with heat response and salinity adaptation. This study could provide valuable genomic resources and information for further research on the molecular evolution, genetic breeding, biological function and evolutionary adaptation of C. ariakensis. [ABSTRACT FROM AUTHOR]

Estuarine and coastal ecosystems are often considered vulnerable due to the complex biogeochemical processes and the human disturbances through a variety of pollution. Among environmental contaminants, heavy metals in estuarine and coastal ecosystems have been of increasing concern in environmental conservation. Long-term exposure to heavy metal contamination, mainly through food and water, could be harmful to human health. It is therefore critical to understand the quantitative comparisons and combined effects of different heavy metals in common seafood species, such as oysters. This work studied the long-term spatiotemporal trends and health risk assessment of oyster arsenic (As), cadmium (Cd), copper (Cu), mercury (Hg), lead (Pb), and zinc (Zn) levels in the coastal waters of northern South China Sea. Cultured oysters (Crassostrea rivularis) from 23 estuaries and harbors in the coastal areas of northern South China Sea in 1989–2015 were analyzed for the spatiotemporal trends of the six heavy metal levels. Metal pollution index (MPI), target hazard quotient (THQ), and hazard index (HI) were used for quantifying the exposure of the six heavy metals to human health through oyster consumption. Principal component analysis (PCA) was used for assessing the relative importance of the six metals in oyster heavy metal distribution patterns in the northern South China Sea. Overall, the As, Cd, Cu, Hg, Pb, and Zn levels in oysters from the northern South China Sea generally declined from 1989 to 2015, stayed relatively high (MPI = 2.42–3.68) during 1989–2000, gradually decreased since 2000, and slightly increased after 2010. Oyster heavy metal levels were highest in the Pearl River Estuary (MPI = 1.20–5.52), followed by west Guangdong and east Guangdong, Guangxi, and Hainan coastal waters. This pattern is probably because economics and industry around the Pearl River Estuary have been growing faster than the other areas of this work in the recent two decades, and it should be taken as a hotspot for the monitoring of seafood safety in southern China. Principal component analysis indicated that Cu, Zn, and Cd were the most important metals in the long-term distributions of oyster heavy metal levels in the northern South China Sea. Health risk assessment suggested that the risk of the six heavy metals exposure through oyster consumption were relatively high during 1989–2005 (THQ = 1.01–5.82), significantly decreased since 2005 (THQ < 1), and slightly increased after 2010. [ABSTRACT FROM AUTHOR]

Understanding the roles of genetic divergence and phenotypic plasticity in adaptation is central to evolutionary biology and important for assessing adaptive potential of species under climate change. Analysis of a chromosome-level assembly and resequencing of individuals across wide latitude distribution in the estuarine oyster (Crassostrea ariakensis) revealed unexpectedly low genomic diversity and population structures shaped by historical glaciation, geological events and oceanographic forces. Strong selection signals were detected in genes responding to temperature and salinity stress, especially of the expanded solute carrier families, highlighting the importance of gene expansion in environmental adaptation. Genes exhibiting high plasticity showed strong selection in upstream regulatory regions that modulate transcription, indicating selection favoring plasticity. Our findings suggest that genomic variation and population structure in marine bivalves are heavily influenced by climate history and physical forces, and gene expansion and selection may enhance phenotypic plasticity that is critical for the adaptation to rapidly changing environments. Ao Li, He Dai, and Ximing Guo et al. present a high-quality genome from the estuarine oyster, Crassostrea ariakensis. They highlight the climate impact on population divergence, a suite of genes likely important for adaptation to environments of different temperatures and salinities, and provide further insight into the evolution of this species. [ABSTRACT FROM AUTHOR]

The hybridization experiments have been made between Crassostrea ariakensis and C. gigas several times. However, it is difficult to obtain a large number of hybrid offspring, which becomes the bottleneck of subsequent cross‐breeding. To explore whether the production of hybrid larvae is affected by salinity, we investigated the effects of salinity (16, 20, 24, 28 and 32 psu) on the early activities of artificial hybridization between C. ariakensis and C. gigas at 23–24°C. In this study, the results showed that during artificial insemination, the appropriate salinity of high‐quality gametes in C. ariakensis and C. gigas was 20–32 psu and 24–32 psu respectively. Besides, the fertilization rate of AG (C. ariakensis♀ × C. gigas♂) and GA (C. gigas♀ × C. ariakensis♂) at 24–28 psu was significantly (p <.05) higher than that at 16–20 psu. For incubation, the optimal salinity of embryo incubation was 28 psu, under which all embryos of AG and GA can develop rapidly to the D‐larvae and yielded a high hatching rate. During larval rearing, the larval shell height and survival rate of AG were of no significant difference (p >.05) at all salinities, and the optimum salinity for larval growth and survival of GA was 24–28 psu. These findings can contribute to the increase in the hybrid progeny yield, which can serve as the new resource for genetic improvement of oyster germplasm. [ABSTRACT FROM AUTHOR]

Copyright of South China Fisheries Science is the property of South China Fisheries Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Bivalve habitat restoration is growing in geographic extent and scale globally. While addressing the wide‐scale loss of these biogenic habitats is still a key motivation behind restoration efforts, stakeholders and funders are increasingly drawn to shellfish restoration for the many ecosystem services these habitats provide.There is clear evidence for the provision of ecosystem services from species targeted for restoration in the USA, in particular Crassostrea virginica. Ecosystem services, however, remain largely unquantified or even undescribed for the majority of other species targeted for restoration.A structured review of the literature was undertaken and supplemented by expert knowledge to identify which ecosystem services are documented in the following other bivalve species targeted for restoration: Ostrea edulis, Ostrea angasi, Crassostrea rhizophorae, Perna canaliculus, Modiolus modiolus, Mytilus edulis, Mytilus platensis, Crassostrea gigas, Ostrea denselamellosa, Crassostrea ariakensis, and Crassostrea sikamea.Key knowledge gaps in quantifying ecosystem services and the ecosystem engineering properties of habitat‐building bivalves contributing to the provision of ecosystem services were identified. Ecosystem services with the potential to be widely applicable across bivalve habitat‐building species were identified.Though there is evidence that many of the ecosystem engineering properties that underpin the provision of ecosystem services are universal, the degree to which services are provided will vary between locations and species. Species‐specific, in situ, studies are needed in order to avoid the inappropriate transfer of the ecosystem service delivery between locations, and to further build support and understanding for these emerging targets of restoration. [ABSTRACT FROM AUTHOR]

Coastal heavy metal pollution has become an important topic for seafood safety and marine environmental protection. Unlike toxic heavy metals such as cadmium or chromium, copper is essential for oysters' growth but can inhibit their immune response to exotic stress when going above normal levels. Oysters with high copper levels can easily accumulate and transfer abnormal amounts of copper to upper trophic levels, and generate health risks for humans. This study investigated the spatiotemporal variability and health risk of copper levels in cultured oysters (Crassostrea rivularis) sampled from 23 harbors, bays, or estuaries along the northern South China Sea during 1989–2015. Overall, oyster copper concentrations in the study area ranged from 0.9 to 1897.0 μg/g wet weight with a mean of 210.0 (± 143.6) μg/g and a median of 89.3 μg/g. Although oyster copper levels in the southern China provinces of Guangdong, Guangxi, and Hainan showed an overall decrease during 1989–2015, they stayed relatively low since 1996 and increased slightly after 2010. Oyster copper levels in Guangdong were significantly higher than in Hainan and Guangxi. In Guangdong, oyster copper levels were highest in the Pearl River Estuary, followed by west Guangdong and east Guangdong. The health risk of copper exposure through oyster consumption increased in 2011–2015 compared with in 2006–2010. It is recommended that the human daily intake of cultured oysters in the study area should be reduced by half to minimize copper exposure. This study suggested that copper is one of the most important heavy metal contaminants in coastal and estuarine ecosystems of the northern South China Sea. [ABSTRACT FROM AUTHOR]

Copyright of South China Fisheries Science is the property of South China Fisheries Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Despite the benefits of shellfish consumption, the bioaccumulation of heavy metals in shellfish can endanger consumer's health. The consumption of seafood in Shenzhen (a fast-developing metropolis in China) has received more and more attention. Arsenic (As), cadmium (Cd), copper (Cu), mercury (Hg), and lead (Pb) in ten common shellfish species and associated health risks were analyzed for Shenzhen's consumers by evaluating estimated weekly intake (EWI), non-carcinogenic and carcinogenic health risks to children, adolescents, and adults. In this study, 50 shellfish samples were collected in total. The results showed that the levels of inorganic arsenic (iAs) in Babylonia areolata exceeded the maximum permissible limit set by the food safety guidelines (0.5 mg/kg), while other elements were below the limit in the present guidelines (Ministry of Health of the People's Republic of China, GB 2762-2012). EWI values of children, adolescents, and adults were all lower than provisional tolerable weekly intakes (PTWIs) of all shellfish species. The analysis of total target hazard quotients (TTHQ) showed that the ingested B. areolata in children, adolescents, and adults were all at non-carcinogenic risks; the consumption of Argopecten irradians and Chlamys farreri would pose non-carcinogenic risks for children only. In all age groups, the consumption of A. irradians, B. areolata, C. farreri, and Crassostrea ariakensis would lead to lifetime cancer risk due to Cd bioaccumulation, with toxicity of Pb and iAs to be acceptable and negligible. [ABSTRACT FROM AUTHOR]

To understand the environmental adaptations among sessile bivalves lacking adaptive immunity, a series of analyses were conducted, with special emphasis on the widely distributed C. ariakensis. Employing Pacbio sequencing and Hi-C technologies, whole genome for each of a C. ariakensis (southern China) and C. hongkongensis individual was generated, with the contig N50 reaching 6.2 and 13.0 Mb, respectively. Each genome harbored over 30,000 protein-coding genes, with approximately half of each genome consisting of repeats. Genome alignment suggested possible introgression between C. gigas and C. ariakensis (northern China), and re-sequencing data corroborated this result and indicated significant gene flow between C. gigas and C. ariakensis. These introgressed candidates, well-represented by genes related to immunity and osmotic pressure, may be associated with environmental stresses. Gene family dynamics modeling suggested immune-related genes were well represented among the expanded genes in C. ariakensis. These outcomes could be attributed to the spread of C. ariakensis. [ABSTRACT FROM AUTHOR]

The antioxidant and aphrodisiac properties of oyster meat (OM) and its hydrolysates by alcalase (OMA) were compared. The results showed that OMA displayed a higher antioxidant activity than OM with or without gastrointestinal digestion. Furthermore, the study reported that oral administration of OM or OMA could induce aphrodisiac activities and consequently enhance the sexual behavior in normal male mice, at a dose of 250 mg/kg. Additionally, OMA also exhibited better antioxidant activity in vivo than OM by improving the activities of the endogenous cellular antioxidant enzymes and decreasing the MDA levels, which may be helpful in improving the sexual function. These results indicated that oysters' could be a potential functional ingredient with antioxidant and aphrodisiac activities, and the activities could be improved by alcalase hydrolysis. Unlabelled Image • OMA displayed higher antioxidant activity both in vitro and in vivo. • OMA could improve the sexual function of normal male mice more efficiently. • The antioxidant activity may contribute to the aphrodisiac property. [ABSTRACT FROM AUTHOR]

[Display omitted] • Bioavailability of two typical shellfish species were performed using in vitro method. • Bioavailability of Zn and Cd in cooked C. rivularis samples both exceeded 82%. • Bioavailability of Zn and Cd in cooked P. undulata samples both exceeded 83%. • The intake of Cd from both shellfish species is safety. Dynamic extraction of edible tissues of Jinjiang oyster (Crassostrea rivularis) and Flower clam (Paphia undulata) was conducted using a fully biomimetic digestion (in vitro) method. The impact of different cooking methods on the bioavailability of Zn and Cd in the edible shellfish tissues was analyzed, and the human health risk of Zn and Cd was evaluated. The results show that the gastric biomimetic extractions of Zn and Cd in unheated samples of C. rivularis and P. undulata were higher than those in the intestinal biomimetic extraction. The extraction patterns of cooked samples were consistent with those of raw samples. The bioavailability of Zn and Cd in cooked samples of C. rivularis was 94.9% and 82.5%, respectively, indicating increased Zn bioavailability but decreased Cd bioavailability compared to the raw samples. The bioavailability of Zn and Cd in cooked samples of P. undulata was 85.1% and 83.0%, respectively, both of which decreased compared to the raw samples. Consumption of C. rivularis can provide 21.0% to 34.2% of the daily required Zn intake, while consumption of P. undulata can provide 3.8% to 6.4%. The intake of Cd from both shellfish species is below the monthly tolerable intake recommended by FAO/WHO. Consuming cooked C. rivularis can increase the intake of Zn and decrease the intake of Cd. [ABSTRACT FROM AUTHOR]

Regulatory variants in gene expression serve as bridges linking genetic variation and phenotypic plasticity. Environmental conditions typically influence the effects of regulatory variants on phenotypic plasticity; however, such genotype-by-environment interactions (G × E) are poorly understood. This study aimed to investigate the genetic basis of G × E in estuarine oyster (Crassostrea ariakensis), which is an important model animal for studying environmental adaption owing to its high plasticity and large intraspecific divergence. Genome-wide mapping of expression quantitative trait loci (eQTLs) for 23 environmental adaptive genes was performed for 256 estuarine oysters. We identified 1194 eQTL single nucleotide polymorphisms (eSNPs), including 433 cis -eSNPs in four genes and 722 trans -eSNPs in eight genes. The expression variation explanation of cis -eSNPs (9.95%) was significantly higher than that of trans -eSNPs (9.15%). We specifically showed cis- and trans-eSNPs with high linkage disequilibrium (LD) for Traf7 , Slc6a5 , Ggt , and Dap3. For example, we identified a cis -regulatory LD block containing 68 cis -eSNP and a trans -regulatory LD block, including 20 trans -eSNPs in Traf7. A high proportion (85%) of 40 vital eSNPs exhibited significant G × E effects. We identified crossing and nonparallel interactions of G × E, with the tag cis -eSNPs of Baat and Slc6a5 as representatives. Our results indicated that cis -eQTLs are highly conserved. This study provides insights into the understanding of adaptive evolutionary mechanisms and phenotypic response prediction to variable environments, as well as the genetic improvement for superior adaptive traits for genetic resource conservation and aquaculture. • We identified trans and cis regulatory variants and built potential regulatory networks. • Cis -eSNPs had larger contributions to gene expression plasticity and are conserved in different environments. • Trans -eSNPs are more abundant than cis -eSNPs and usually environment-specific. • A large proportion of eSNPs exhibited significant G × E effects. • Crossing and nonparallel G × E patterns were identified. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Hemocyte parameters of oysters were determined by FC in vitro and in vivo. • Synergistic effects of Cu and low pH on oyster hemocyte were observed. • Cu exerted a stronger influence on hemocyte parameters than low pH. • Carry-over impacts of Cu and low pH were found during the recovery period. Abstract With the development of industry and agriculture, the metal pollutants (e.g., Cu) are inevitably released into the aquatic environment. In addition, ocean acidification (OA) as a major environmental stress is affecting marine organisms. In this study, we investigated the hemocyte responses of the estuarine oyster Crassostrea rivularis exposed to six combinations of two pH levels (8.1 and 7.7) and three Cu concentrations (0, 10 and 50 μg/l) using flow cytometry in vitro and in vivo. In both experiments, Cu and low pH jointly affected the hemocyte parameters of oyster. High Cu exposure resulted in decreased total hemocyte count (THC), esterase activity (EA) and lysosomal content (LC) and increased hemocyte mortality (HM), phagocytosis activity (PA) and reactive oxygen species (ROS) production, especially under low pH conditions. The immune suppression of metal-exposure was more significant than low pH exposure with a 28-d experimental period in oysters. A slight recovery of the immune parameters was observed in THC, HM, PA, ROS and LC. During the depuration period, the modulatory effects of pH were still obvious. In addition, carry-over effects of high Cu and low pH were still observed. Overall, our results showed that copper and low pH weaken immune functions of hemocyte in oysters, with synergistic effects. This work provides new evidence of sublethal negative effects of metals on marine animals under global change scenarios, and copper likely leads to reduced fitness of oysters under low pH conditions. [ABSTRACT FROM AUTHOR]

Abstract: Crude polysaccharide was prepared from Crassostrea rivularis by 30% (w/v) potassium hydroxide solution at 90°C for 120 min. Three fractions (OG1, OG2, and OG3) were purified by DEAE‐52 cellulose and Sepharose 2B gel column chromatography. The chemical structures were determined using gas chromatography (GC), high‐performance gel permeation chromatography (HPGPC), Fourier‐transform infrared (FT‐IR) spectroscopy, and 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. The results indicated that OG1 was composed of rhamnose and little mannose (8.71%), the ratio of Rha: Gal: Xyl: Fuc in OG3 were 14:5.5:3:1. And their average molecular weights (Mw) were about 1.66 × 106 and 2.33 × 106 Da, respectively. OG2 was composed only of glucose (98.23%), which means it was glycogen. OG2 was consisted mainly of →4)‐ α‐D‐Glc‐(1→, with the branch chain every 6.5 glucose residues on average, which is →4,6)‐α‐D‐Glc‐(1→ and trace amount of α‐D‐Glc‐(1→ branched units. The Mw was 2.27 × 106 Da. It provides the bases for the bioactivity research. [ABSTRACT FROM AUTHOR]

With the rapid development of mariculture in potentially contaminated regions in China and the world, food safety, is a growing concern. To evaluate heavy metals and their associated health risks in the cultivated oyster Crassostrea rivularis, the concentrations of heavy metals (Cd, Cr, Pb, Zn) in oysters and water/sediment were examined in a typical mariculture environment (Kaozhou Bay, South China Sea). Trends in the seasonal dynamics of heavy metals in oysters revealed a potential synergistic effects among the concentrations of Cd, Cr, and Zn; trends associated with Pb were less clear, although the ability of oysters to bioaccumulate and depurate Pb was excellent. Bioconcentration factors (BCF) indicated that C. rivularis has a strong ability to accumulate heavy metals, and the BCF was the highest for Zn (2.32 × 105), followed by Cd (6.84 × 104), Pb (2.77 × 104) and Cr (1.23 × 103) through the four seasons. Results showed that Cd concentrations in oysters could pose a risk to human health (HQ > 1). This study, therefore, suggests that there are potential human health risks due to heavy metal exposure through the consumption of C. rivularis from mariculture zones in South China Sea. [ABSTRACT FROM AUTHOR]

The growth of Vibrio parahaemolyticus (V. parahaemolyticus) in oysters during postharvest storage increases the possibility of its infection in humans. In this work, to investigate the growth or survival profiles in different media, pathogenic V. parahaemolyticus in APW, Jinjiang oyster (JO, Crassostrea rivularis) slurry, and live JO were studied under different temperatures. All the strain populations were counted through our double-layer agar plate (DLAP) method. In APW, the pathogenic V. parahaemolyticus showed continuous growth under 15, 25, and 35°C, while a decline in behavior was displayed under 5°C. The similar survival trend of pathogenic V. parahaemolyticus in JO slurry and live JO was observed under 5, 25, and 35°C, except the delayed growth or decline profile compared to APW. Under 15°C, they displayed decline and growth profile in JO slurry and live JO, respectively. These results indicate the different sensitivity of pathogenic V. parahaemolyticus in these matrices to temperature variation. Furthermore, nonpathogenic V. parahaemolyticus displayed little difference in survival profiles when inoculated in live JO under corresponding temperatures. The results indicate that inhibition or promotion effect could be regulated under different storage temperature for both pathogenic and nonpathogenic strains. Besides, the DLAP method showed the obvious quickness and efficiency during the bacteria count. [ABSTRACT FROM AUTHOR]

A polyclonal antibody specific to an egg protein of Suminoe oyster Crassostrea ariakensis was previously developed in our laboratory to assess the reproductive life cycle of the oyster. The present study was undertaken to investigate vitellin of C. ariakensis (CAVt). Vitellin is an essential component of egg proteins in marine invertebrates as it provides energy and nutrients to the embryo and larvae. CAVt was purified from eggs of the oyster using ammonium sulfate precipitation followed by affinity chromatography with Concanavalin A-agarose. Native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate PAGE showed that CAVt is a high molecular weight [532 kiloDaltons (kDa)] protein, with multiple subunits. Similar to other vitellin proteins, it is a phospholipoglycoprotein composed of phospholipids (12.06%), carbohydrates (mannose, 10.08% or glucose, 9.84%), and alkali-labile phosphates (4.16%). Affinity chromatography, enzyme-linked immunosorbent aasay (ELISA) and western blot analysis revealed that CAVt is only present in the ovary, and two subunits of CAVt (72 and 35 kDa) are believed to be incorporated from the hemolymph into the oocyte. The antibody specific to CAVt (anti-CAVt), raised in rabbit, strongly cross reacted with the egg proteins of oyster species and scallops, suggesting that the antigenic epitopes are highly conserved among species. Our results suggest that the anti-CAVt antibody can be used to develop a tool similar to ELISA or western blotting for investigation of the effect of microorganisms on reproduction as well as the effect of chemicals on the endocrine system in C. ariakensis. [ABSTRACT FROM AUTHOR]

Densoviruses have short ssDNA genomes and mainly infect arthropods. To characterize viral nucleic acid in shellfish, oysters ( Crassostrea ariakensis) were analyzed using viral metagenomics. Two large de novo assembled contigs, CaaDV1 and CaaDV2, consisting of nearly complete densovirus genomes (5860 nucleotides (nt) and 4034 nt) with two major ambisense protein coding regions were identified. Several potential non-structural proteins and capsid proteins were encoded by these genomes, but these were divergent from the existing densoviral species. The NS1 protein of the two CaaDVs shared 43.3%~61.5% amino acid identities with the sea star-associated densovirus and cherax quadricarinatus densovirus, with the four species clustering by phylogenetic analysis. This is the first report of densovirus detection in shellfish, increasing the potential host range of densoviruses and the genetic diversity of the genus Ambidensovirus. [ABSTRACT FROM AUTHOR]

This study investigates the nutritional properties of edible oysters cultivated in Hong Kong (i.e. Deep Bay) to determine their nutritional values compared with other foodstuffs (e.g. land and aquatic-based foods). A total of ∼300 edible oysters were harvested and studied from aquaculture areas in Hong Kong. The Hong Kong oyster (Crassostrea hongkongensis) contained the highest concentrations (up to 32%) of polyunsaturated fatty acids (PUFAs), when compared to the Portuguese (Crassostrea angulata) and the Suminoe (Crassostrea ariakensis) oyster species. The C. hongkongensis also exhibited the best meat yield (∼28%) compared to oyster species from other regions (e.g., Crassostrea gigas). The C. ariakensis showed the highest concentrations (µg/g dwt.) of macro-and micro nutrients, e.g. Mg (∼3600), Na (∼22000), Zn (∼5000); compared to C. hongkongensis and C. angulata. Essential amino acids (EEA; mg/100 g wwt.) were highest in C. angulata (3439), followed by C. ariakensis (3330) and leaving C. hongkongensis (2619) as the poorest species in EAA. The comparison of the nutritional profile of oyster species cultivated in Hong Kong with different seafood species and other foodstuffs showed that oysters are nutrient-rich (or superfoods) shellfish species, which contribute with beneficial compounds (e.g. PUFAs, EAAs) and elements (e.g. Cu, Zn) for a human healthy diet. • The C. hongkongensis contained the highest concentration of PUFAs among oysters. • The C. hongkongensis has the best meat yield compared to oysters from other regions. • The C. ariakensis showed the highest concentrations of macro-and micro nutrients. • EEA were highest in C. angulata , following by C. ariakensis and C. hongkongensis. • Consumption of Hong Kong edible oysters can contribute positively for human health. [ABSTRACT FROM AUTHOR]

Oysters are a keystone species in estuaries and intertidal zones and have important economic and ecological significance; however, worldwide, the dominant aquaculture oyster species suffers from mass mortality owing to anthropogenic impacts and rapid climate change. As a possible solution, inter/intraspecies hybridization is a feasible way to breed new germplasms with genetic improvement and high environmental adaptability. Suminoe oysters (Crassostrea ariakensis) are broadly distributed in the estuaries of China and have evolved strong genetic divergence between the northern and southern populations; therefore, they are ideal candidates to explore intraspecies heterosis and the associated molecular mechanisms. In this study, we performed reciprocal hybridization and examined fitness traits in oysters cultured in both northern and southern habitats. Reciprocal hybrids performed better than inbred populations in both environments, indicating heterosis in Suminoe oysters. Hybrids reared in maternal habitats showed faster growth, higher field survival, and greater aerobic metabolism than alternative hybrid oysters under the same conditions. The transcriptional analysis identified modules related to heterosis and maternal effect, while genes responsible for galactose metabolism, glycolysis/gluconeogenesis, and the tricarboxylic acid cycle were enriched and upregulated in hybrids, especially those in maternal environments. Our findings reveal that accelerated aerobic energy metabolism, which may result from mitonuclear coadaptation, underlies the molecular basis of heterosis and maternal effect in the hybridization of Suminoe oysters. Thus, we provide important insights for the aquaculture management of marine species, revealing that selecting appropriate environments (maternal habitats) is critical for heterosis in cross-breeding. • Hybrids from divergent oyster populations showed heterosis in growth and survival. • Hybrids in maternal habitat exhibited better performance indicating maternal effect. • Accelerated aerobic metabolism underlies heterosis and maternal effect in oyster. • Our findings propose better hybridization breeding of hybrids in maternal habitat. [ABSTRACT FROM AUTHOR]

The estuarine oyster (Crassostrea ariakensis), a species native to Asia, is widespread throughout the estuarine region along the northern and southern coasts of China and has important ecological and economic values. Owing to runoff dropping and environmental threats caused by climate change and anthropogenic influences, the natural resources of C. ariakensis have persistently declined. Therefore, breeding oysters with high adaptability, which requires efficient and accurate genotyping tools, is beneficial to restore the germplasm resources and aquaculture. However, targeted genotyping tools are poorly developed for this species and even in other aquatic species. Previously, we have detected some selective regions and genes associated with temperature and salinity adaptation using selection signature analysis and found that the upstream regulatory regions of the environment-responsive genes exhibited higher differentiation. In this study, we developed a single nucleotide polymorphism (SNP) targeted genotyping tool for C. ariakensis , which includes 30 K SNPs located in previously identified target regions. The target genomic regions were enriched by hybridization capture using double-stranded DNA (dsDNA) biotin-labeled probes, and sequenced with high depth. A total of 24,541 (81.8%) high-quality SNPs were successfully genotyped in 655 samples. Compared to the genotypes obtained from previous genome-wide resequencing, 74.2% of the SNPs had a consistency rate of 1, with an average consistency rate for each sample of 97.8%. Population structure analysis showed three divergent populations (northern, southern, and middle) on the coast of China, consistent with those constructed using SNPs derived from whole-genome resequencing data. Most of the 30 K SNPs were located in the intergenic and regulatory regions, whereas 7% were located in the coding region. These high-quality SNPs were located in 953 genes, including 100 heat-response and 348 high-salinity-responsive genes. Therefore, this panel can be used to evaluate the adaptive divergence between populations, and causative SNPs related to temperature and salinity adaptation in the linkage and association analysis, providing robust support for the molecular breeding of estuarine oyster. To the best of our knowledge, this study proposes the first high-throughput SNP-targeted genotyping tool for studying the genetic basis of environmental adaptation in oyster, and providing insights into the development of targeted SNP genotyping tools for the adaptation study in other aquatic species under climate changes. • We developed a 30 K targeted genotyping tool for Crassostrea. Ariakensis. • 24,541 high-quality SNPs were successfully genotyped in 655 samples. • These high-quality SNPs were located in environment-responsive genes. • 74.2% of the SNPs had a consistency rate of 1 with resequencing. Population structure analysis showed three divergent populations on the coast of China • This study could be used to dissect the genetic basis of environmental adaptation. [ABSTRACT FROM AUTHOR]

Cyclophilins (CyPs) are a family of proteins that bind the immunosuppressive agent cyclosporin A (CsA) with high-affinity and belong to one of the three superfamilies of peptidyl-prolyl cis-trans isomerases (PPIase). In this report, three cyclophilin genes (Ca-CyPs), including Ca-CyPA, Ca-CyPB and Ca-PPIL3, were identified from oyster, Crassostrea ariakensis Gould in which Ca-CyPA encodes a protein with 165 amino acid sequences, Ca-CyPB encodes a protein with 217 amino acid sequences and Ca-PPIL3 encodes a protein with 162 amino acid sequences. All of the three Ca-CyPs genes contain a typical CyP-PPIase domain with its signature sequences and Ca-CyPB contains an N-signal peptide sequences. Tissue distribution study revealed that Ca-CyPs were ubiquitously expressed in all examined tissues and the highest levels were observed in hemocytes. RLO incubation upregulated the mRNA expression levels of Ca-CyPs, indicating that three Ca-CyPs might be involved in oyster immune response against RLO infection. [ABSTRACT FROM AUTHOR]

The impact of coastal pollution was studied using edible oysters, Crassostrea rivularis as an indicator at two sites viz., North Wandoor (NW) and Phoenix Jetty (PJ) in Port Blair, Andaman. The hydrographic parameters showed that nitrite, nitrate and phosphate concentration were less and dissolved oxygen were more at NW compared to PJ. The oysters were collected from the study sites and biochemical, microbial, mineral profiles and ATPase activities were estimated. ATPase activity was inhibited in the gill tissue of oysters ( p < 0.05) of PJ sample. Total microbial load in the water and oyster, and coliform bacteria (MPN) in the water were significantly ( p < 0.05) higher at PJ compared to the NW. There was no significant difference ( p > 0.05) in the mineral profile of water collected from both the sites. However, calcium and magnesium were more in the oysters collected from NW ( p < 0.05), and Cu, Zn and Cd were more in PJ samples ( p < 0.05). [ABSTRACT FROM AUTHOR]

A central composite experimental design and response surface method were used to investigate the combined effects of water temperature (18-34°C) and copper ion concentration (0.1-1.5 mg/L) on the catalase (CAT) activity in the digestive gland of Crassostrea ariakensis. The results showed that the linear effects of temperature were significant ( P<0.01), the quadratic effects of temperature were significant ( P<0.05), the linear effects of copper ion concentration were not significant ( P>0.05), and the quadratic effects of copper ion concentration were significant ( P<0.05). Additionally, the synergistic effects of temperature and copper ion concentration were not significant ( P>0.05), and the effect of temperature was greater than that of copper ion concentration. A model equation of CAT enzyme activity in the digestive gland of C. ariakensis toward the two factors of interest was established, with R, Adj. R and Pred. R values as high as 0.943 7, 0.887 3 and 0.838 5, respectively. These findings suggested that the goodness of fit to experimental data and predictive capability of the model were satisfactory, and could be practically applied for prediction under the conditions of the study. Overall, the results suggest that the simultaneous variation of temperature and copper ion concentration alters the activity of the antioxidant enzyme CAT by modulating active oxygen species metabolism, which may be utilized as a biomarker to detect the effects of copper pollution. [ABSTRACT FROM AUTHOR]

Abstract: 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 R 2=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. [Copyright &y& Elsevier]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract: 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. [Copyright &y& Elsevier]

Highlights: [•] We surveyed Marteilioides infection in marine bivalves using histology and PCR. [•] Marteilioides infection was also confirmed in the male oysters and clams. [•] The nested PCR was much more sensitive than histology in the detection of Marteilioides. [•] Marteilioides found in Manila clam is a considered to be a new Marteilioides sp. [Copyright &y& Elsevier]

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 reproductiveC. ariakensisfrom 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 reproductiveC. ariakensisthat 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 sterileC. ariakensiswould 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. [ABSTRACT FROM AUTHOR]

Background: Crassostrea ariakensis Gould is a representative bivalve species and an economically important oyster in China, but suffers severe mortalities in recent years that are caused by rickettsia-like organism (RLO). Prevention and control of this disease is a priority for the development of oyster aquaculture. It has been proven that mammalian HMGB (high mobility group box) can be released extracellularly and acts as an important pro-inflammatory cytokine and late mediator of inflammatory reactions. In vertebrates, HMGB's antibody (anti-HMGB) has been shown to confer significant protection against certain local and systemic inflammatory diseases. Therefore, we investigated the functions of Ca-HMGB (oyster HMGB) and anti-CaHMGB (Ca-HMGB's antibody) in oyster RLO/LPS (RLO or LPS)-induced disease or inflammation. Methodology/Principal Findings: Sequencing analysis revealed Ca-HMGB shares conserved structures with mammalians. Tissue-specific expression indicates that Ca-HMGB has higher relative expression in hemocytes. Significant continuous upregulation of Ca-HMGB was detected when the hemocytes were stimulated with RLO/LPS. Recombinant Ca-HMGB protein significantly up-regulated the expression levels of some cytokines. Indirect immunofluorescence study revealed that Ca-HMGB localized both in the hemocyte nucleus and cytoplasm before RLO challenge, but mainly in the cytoplasm 12 h after challenge. Western blot analysis demonstrated Ca-HMGB was released extracellularly 4-12 h after RLO challenge. Anti- CaHMGB was added to the RLO/LPS-challenged hemocyte monolayer and real-time RT-PCR showed that administration of anti-CaHMGB dramatically reduced the rate of RLO/LPS-induced up-regulation of LITAF at 4-12 h after treatment. Flow cytometry analysis indicated that administration of anti-CaHMGB reduced RLO/LPS-induced hemocyte apoptosis and necrosis rates. Conclusions/Significance: Ca-HMGB can be released extracellularly and its subcellular localization varies when stimulated with RLO. Ca-HMGB is involved in oyster immune reactions and functions as a pro-inflammatory cytokine. Anti-CaHMGB can significantly suppress RLO/LPS-induced inflammatory responses and hemocyte necrosis and apoptosis, suggesting that Ca-HMGB is a potential target to prevent and control RLO/LPS-induced disease or inflammation. [ABSTRACT FROM AUTHOR]

Abstract: We identified a tetraspanin family member gene, named Ca-TSP, in the oyster Crassostrea ariakensis and found that the transcription profiles of Ca-TSP were variable in the oyster hemocytes. Three distinct patterns of variation of Ca-TSP were observed. Using immunofluorescence and immunoelectron microscopy, we show that Ca-TSP was present in granules and in vesicular structures of the oyster hemocyte. Sequence analysis, structural features and immunogold electron microscopy showed that Ca-TSP is an integral membrane glycoprotein of granules of hemocyte and may be a novel CD63-like gene of the tetraspanin family of molluscs. The gene expression analysis of Ca-TSP using isolated oyster hemocytes, was done following challenge of the oysters with LPS and Poly I:C. The Ca-TSP mRNA levels increased in hemocytes in the first 12 h after LPS and Poly I:C stimulation, and decreased after the addition of H2O2. Western blot analysis using anti-Ca-TSP antibody indicated that gene expression and protein levels were similar. The recombinant Ca-TSP was found to significantly inhibit hemocytes aggregation. Our results suggested that Ca-TSP participates in the innate immunity of the oyster. [Copyright &y& Elsevier]

Abstract: A SMARTer™ cDNA library of hemocyte from Rickettsia-like organism (RLO) challenged oyster, Crassostrea ariakensis Gould was constructed. Random clones (400) were selected and single-pass sequenced, resulted in 200 unique sequences containing 96 known genes and 104 unknown genes. The 96 known genes were categorized into 11 groups based on their biological process. Furthermore, we identified and characterized three complement-related fragments (CaC1q1, CaC1q2 and CaC3). Tissue distribution analysis revealed that all of three fragments were ubiquitously expressed in all tissues studied including hemocyte, gills, mantle, digestive glands, gonads and adductor muscle, while the highest level was seen in the hemocyte. Temporal expression profile in the hemocyte monolayers reveled that the mRNA expression levels of three fragments presented huge increase after the RLO incubation at 3 h and 6 h in post-challenge, respectively. And the maximal expression levels at 3 h in post-challenge are about 256, 104 and 64 times higher than the values detected in the control of CaC1q1, CaC1q2 and CaC3, respectively. [Copyright &y& Elsevier]

Oysters can create reefs that provide habitat for associated species resulting in elevated resident abundances, lower mortality rates, and increased growth and survivorship compared to other estuarine habitats. However, there is a need to quantify trophic relationships and transfer at created oyster reefs to provide a better understanding of their potential in creating suitable nekton habitat. Stable isotope analyses (δC and δN) were conducted to examine the organic matter sources and potential energy flow pathways at a created intertidal oyster ( Crassostrea ariakensis; hereinafter, oyster) reef and adjacent salt marsh in the Yangtze River estuary, China. The δC values of most reef-associated species (22 of 37) were intermediate between those of suspended particle organic matter (POM) and benthic microalgae (BMI), indicating that both POM and BMI are the major organic matter sources at the created oyster reef. The sessile and motile macrofauna colonizing the reef make up the main prey of transient nekton (e.g., spotted sea bass, Asian paddle crab, and green mud crab), thus suggesting that the associated community was most important in supporting higher trophic levels as opposed to the direct dietary subsidy of oysters. The created oyster reef consistently supported higher trophic levels than the adjacent salt marsh habitat due to the dominance of secondary consumers. These results indicate that through the provision of habitat for associated species, created oyster reefs provide suitable habitat and support a higher average trophic level than adjacent salt marsh in the Yangtze River estuary. [ABSTRACT FROM AUTHOR]

Crassostrea sikamea and C. ariakensis are two oysters from Asia that are sympatric and differ slightly in their environmental preference. They can hybridize in one direction, although the fertilization level and survival of hybrid larvae to spat are low. To understand the possible effects of salinity on cross-fertilization and the survival of hybrid larvae, we conducted hybridization experiments at four salinities (20, 25, 30 and 35 ppt). At all salinities, about 10% of C. sikamea eggs were fertilized by C. ariakensis sperm, while the reciprocal cross completely failed to fertilize. Survival of fertilized eggs to D-stage in the hybrid cross (C. sikamea female×C. ariakensis male, SA) was significantly (P<0.05) higher at 35 ppt than at lower salinities, while the effect of salinity was not significant in two pure-species crosses. High salinity negatively affected larval survival to day 7 in C. ariakensis and the hybrid cross. Larval growth in all groups was negatively correlated with salinity (P<0.01). Larval shell height at day 7 as well as daily increment was significantly smaller for hybrid than for purebred larvae (P<0.05). These results indicate that hybrid larvae prefer high salinity during the early stages of development but low salinity at later stages. The low and one-directional fertilization represents a pre-zygote barrier to hybridization, which may play a role in sympatric speciation. [ABSTRACT FROM AUTHOR]

Abstract: PCBs equivalent to Aroclor 1242 and 1254 in soft tissues of oysters Crassostrea rivularis from the Guangdong coast, China, were measured using a GC-ECD. PCB concentrations (ngg−1 d.w.) ranged from 30 to 2040 with an average of 315 in 2003–2007, and 0.35–1.43 with an average of 0.56 in 1989–1991. Annual averages were 0.61, 0.65, 313, 290 and 342ngg−1, respectively in 1989, 1991, 2003, 2006 and 2007. In east Guangdong, the Pearl River Estuary, and west Guangdong, regional means were 444, 273, 194ngg−1, respectively in 2003–2007, and 0.43, 0.78, 0.38ngg−1, respectively in 1989–1991, indicating PCB levels in oysters have risen greatly by ratios of 350–1032 compared with earlier data. PCB sources may derive from older transformers, printing materials and pressworks. Fortunately, current residual PCBs in the oysters were 0.004–0.253mgkg−1 (w.w.), still below hygienic criteria of China and developed countries. [Copyright &y& Elsevier]

The costs and benefits of non-native introductions as a restoration tool should be estimated prior to any action to prevent both undesirable consequences and waste of restoration resources. The suggested introduction of non-native oyster species, Crassostrea ariakensis, into Chesapeake Bay, USA, provides a good example in which the survival of non-native oysters may differ from that of native oysters, Crassostrea virginica, during the larval stage. Experiments were conducted to compare the predation vulnerability of native and non-native oyster larvae to different predator types (visual vs. non-visual, benthic vs. pelagic). The results suggest that the non-native larvae are more vulnerable to visual and non-visual pelagic predators. Although vulnerability was similar for larvae exposed to benthic non-visual predators, the consumption of one non-native strain was higher than the consumption of native C. virginica larvae. When vulnerability data are combined with predator feeding rates, the predation mortality for non-native larvae in the wild can be much higher than for native larvae. Small changes in larval mortality rates can yield large changes in total larval delivery to the reef for settlement, so these differences among species may contribute to differences in settlement success. These results provide an example of how a comprehensive examination of the perceived benefits of non-native introductions into complex ecosystems can provide important information to inform management decisions. [ABSTRACT FROM AUTHOR]