Effects of chronic cold stress on tissue structure, antioxidant response, and key gene expression in the warm-water bivalve Chlamys nobilis.

As ectothermic invertebrates, mollusks are regarded as good environmental indicator species for determining the adverse effects of climate change on marine organisms. In the present study, the effects of cold stress on the tissue structure, antioxidant activity, and expression levels of genes were evaluated in the warm-water noble scallop Chlamys nobilis by simulating natural seawater cooled down during winter from 17 °C to 14 °C, 12 °C, 10 °C, and 9 °C. Firstly, the gill was severely damaged at 10 °C and 9 °C, indicating that it could be used as a visually indicative organ for monitoring cold stress. The methylenedioxyamphetamine (MDA) content significantly increased with the temperatures decreasing, meanwhile, the antioxidant enzyme activities superoxide dismutase (SOD) and catalase (CAT) showed a similar pattern, suggesting that the scallop made a positive response. More importantly, 6179 genes related to low temperatures were constructed in a module-gene clustering heat map including 10 modules. Furthermore, three gene modules about membrane lipid metabolism, amino acid metabolism, and molecular defense were identified. Finally, six key genes were verified, and HEATR1, HSP70B2, PI3K, and ATP6V1B were significantly upregulated, while WNT6 and SHMT were significantly downregulated under cold stress. This study provides a dynamic demonstration of the major gene pathways' response to various low-temperature stresses from a transcriptomic perspective. The findings shed light on how warm-water bivalves can tolerate cold stress and can help in breeding new strains of aquatic organisms with low-temperature resistance.

Genome-wide identification of STATs and analysis of their role in sex determination in Pacific oysters (Crassostrea gigas).

STAT (signal transducer and activator of the transcription) proteins, are a group of highly conserved transcription factors and fundamental components of the JAK-STAT signaling pathway. They play crucial roles in a variety of biological processes, such as immunity, proliferation, differentiation, and growth. However, little information is known regarding their role in gonad development and sex determination in mollusks. In this study, we identified 3 STAT genes in Pacific Oyster Crassostrea gigas. Phylogenetic analysis showed that STATs from mollusks were highly conserved, and most of them had four identical motif regions, except for the STAT1 and STAT3 predicted sequences from Crassostrea hongkongensis. Tissue expression analysis indicated CgSTAT1 had a high expression level in most tissues, while CgSTAT3 had a low expression level in most tissues. Expression analysis of early developmental stages showed CgSTAT1 had a higher expression level from egg to D shaped larva and a lower expression level in subsequent stages. In contrast CgSTAT1, CgSTAT2 had a reverse expression pattern. Expression analysis of different developmental stages of diploid gonads indicated that CgSTAT1 had a higher expression level at the S1 and S3 stages relative to the S2 stage in females, while in males the S3 stage had a higher expression than than the S2 stage. The expression level of CgSTAT1 between diploids and triploids in females differed significantly, but there were no significant differences in males. Expression of CgSTAT2 differed significantly between diploid and triploid males. These data suggest an important role for STATs in sex differentiation in diploid and triploid oysters. Our study is the first to explore the role of STATs in sex differentiation and gonadal development in oysters, and will help us better understand the molecular mechanisms of sex differentiation in shellfish.

Effects of high stocking density on growth performance and expression of MyD88, and its temporal expression under the challenge of Vibrio parahaemolyticus in the noble scallop Chlamys nobilis.

High stocking density has been regarded as an adverse factor in bivalve aquaculture. However, its subsequent molecular response to pathogenic bacteria has been little studied. In order to study the question, a novel MyD88 was first cloned using adult noble scallops Chlamys nobilis (CnMyD88), and its tissue distribution was investigated. Then, 1860 juvenile scallops were divided into two groups with two initial densities of high density (200 individuals/layer, HD) and normal density (110 individuals/layer, ND) and in-situ cultured for three months, in which their growth, survival, and the differential expression of CnMyD88 were examined, respectively. Finally, scallops were injected with the Vibrio parahaemolyticus to assess the temporal expression of CnMyD88. As the results show, CnMyD88 cDNA has a full length of 2241 bp and contains an 1107 bp ORF that encodes a 368-derived protein. It was widely expressed in examined tissues with a significantly higher level in hemolymph, intestine, mantle, and gonad than others. Besides, the HD group showed lower growth (0.39 ± 0.05 mm/day) and survival (37.00 ± 8.49%) than the ND group (0.55 ± 0.02 mm/day and 76.82 ± 5.78%). More importantly, the HD group exhibited significantly lower expression levels of CnMyD88 in their examined tissues than the ND group. After V. parahaemolyticus challenging, CnMyD88 had significantly lower expression levels in the scallops from the HD group than that of the scallops from the ND group at 6th, 24th, and 36th. The present results indicated that high stocking density not only made adverse impacts on growth and survival but also may induce immunosuppression in the noble scallop. Therefore, appropriate low stocking density may be worth considering to adopt in scallop aquaculture.

Transcriptome analysis provides insights into the high ability to synthesize fatty acids in "yellow oil" mud crab (Scylla paramamosain).

Yellow oil mud crab (YOC) is a new variant of mud crab (Scylla paramamosain), which was attracted much attention in recent years due to its high level of nutrition. However, the nutritive values and the physiological changes in YOC have not been clearly understood. In this study, we aimed to identify the nutrient compositions (including total carotenoid content (TCC), total lipid content (TLC), total antioxidant capacity (TAC), and fatty acids) and differences in genes related to the biosynthesis of fatty acids using transcriptome analysis in YOC in comparison with those of normal mud crabs. As a result, observations on the morphological characteristics showed that the YOC exhibits a difference in the color of the muscle, gills (orange-yellow), and hemolymph (yellow) compared with the normal female crabs (NFC) (blue or nattier blue). The TCC and TLC (84.96 ± 9.65 µg/g in muscle and 1.39 ± 0.10 µg/mL in hemolymph) or TAC (1.52 ± 0.17 mM in hemolymph) of YOC were higher than that of NFC and normal male crab (NMC). YOC had lower saturated fatty acids, but higher unsaturated fatty acids, as well as the ratio of n-3/n-6 of fatty acids in muscle and hemolymph, compared with those of NFC and NMC. Furthermore, the transcriptome profile revealed that the unigenes in YOC were enriched in the synthesis of n-3 fatty acids. Furthermore, more unigenes related to 'Biosynthesis of unsaturated fatty acids' were identified in muscle and hemocytes, while fewer were in the gonads of YOC. Additionally, the positive (in muscle and hemocytes) and a negative correlation (in gonads) between expressions of unigenes and contents of TLC, TCC, and UFA were found, indicating a better synthesis ability of fatty acids in the muscle and hemocytes of YOC. Overall, compared to NFC and NMC, YOC has higher nutrients and is a better food nutrient source for humans.

In vitro fermentation of Gracilaria lemaneiformis and its sulfated polysaccharides by rabbitfish gut microbes.

This study intended to characterize the Gracilaria lemaneiformis (SW)-derived polysaccharide (GLP) and explore the fermentation aspects of SW and GLP by rabbitfish (Siganus canaliculatus) intestinal microbes. The GLP was mainly composed of galactose and anhydrogalactose (at 2.0:0.75 molar ratio) with the linear mainstay of α-(1 â†’ 4) linked 3,6-anhydro-α-l-galactopyranose and ß-(1 â†’ 3)-linked galactopyranose units. The in vitro fermentation results showed that the SW and GLP could reinforce the short-chain fatty (SCFAs) production and change the diversity and composition of gut microbiota. Moreover, GLP boosted the Fusobacteria and reduced the Firmicutes abundance, while SW increased the Proteobacteria abundance. Furthermore, the adequacy of feasibly harmful bacteria (such as Vibrio) declined. Interestingly, most metabolic processes were correlated with the GLP and SW groups than the control and galactooligosaccharide (GOS)-treated groups. In addition, the intestinal microbes degrade the GLP with 88.21 % of the molecular weight reduction from 1.36 × 105 g/mol (at 0 h) to 1.6 × 104 g/mol (at 24 h). Therefore, the findings suggest that the SW and GLP have prebiotic potential and could be applied as functional feed additives in aquaculture.

Evaluation of the Feasibility of Harvest Optimisation of Soft-Shell Mud Crab (Scylla paramamosain) from the Perspective of Nutritional Values.

Soft-shell crabs have attracted consumers' attention due to their unique taste and nutritional value. To evaluate the feasibility of harvest optimisation of soft-shell mud crabs, the proximate composition, mineral composition, and total carotenoid, amino acid, and fatty acid contents of edible parts of male and female soft-shell mud crabs at different moulting stages were determined and compared from a nutritional value perspective. The results showed that the sex and moulting stages could significantly affect the nutritional values of the edible portions of soft-shell crabs. The female or male soft-shell crabs in the postmoult Ⅰ stage had a much richer mineral element content than that in other moulting stages. The total carotenoid content in female soft-shell crabs was significantly higher than that in male crabs in all moulting stages, while male soft-shell crabs had better performance in amino acid nutrition than female soft-shell crabs. Moreover, it was found that soft-shell crabs in the postmoult Ⅱ stage had significantly higher contents of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), while significantly lower contents of saturated fatty acids (SFA) than those in other stages. The present study will provide a reference basis for the diversified cultivation of soft-shell crabs, and further promote the development of the mud crab industry.

Carotenoid content and composition: A special focus on commercially important fish and shellfish.

Carotenoids are natural pigments that provide many health benefits to living organisms. Although terrestrial plants are the major dietary source of carotenoids for humans, aquatic animals (especially fish and shellfish) are equally important because they are rich in certain important carotenoids lacking in fruits and vegetables. Although extensive research has focused on exploring the carotenoid content and composition in fish and shellfish, this information is poorly organized. This paper reviews the scientific evidence for the carotenoid content and composition in fish and shellfish. It makes serious attempts to summarize the relevant data published on specific research questions in order to improve the understanding of various evidence to clarify the research status of carotenoids in fish and shellfish and defining topics for future studies. From the analysis of published data, it is obvious that most fish and shellfish are rich in complex carotenoids (e.g. astaxanthin, fucoxanthin, fucoxanthinol, lutein). These carotenoids have stronger antioxidant effect, higher efficiency in removing the singlet oxygen and the peroxyl radicals, and have a variety of health benefits. Carotenoid levels in fish and shellfish depend on genotype, climatic conditions of the production area, storage and cooking methods. However, the information of the bioavailability of fish/shellfish carotenoids to human is very limited, which hinders the actual contributions to health. The findings of this study can be used as a guide to select appropriate fish and shellfish as dietary sources of carotenoids, and provide information about potential fish and shellfish species for aquaculture to produce carotenoids to meet part of the growing demand for natural carotenoids.

Identification of a key gene StAR-like-3 responsible for carotenoids accumulation in the noble scallop Chlamys nobilis.

Carotenoids play important roles in living organisms. However, animals cannot synthesize carotenoids by themselves, and they must absorb and accumulate carotenoids from their diets in which some key genes are involved. In present study, a gene named StAR-like-3 was characterized in the noble scallop Chlamys nobilis, and its function was identified using golden scallops with higher carotenoids content and brown scallops with less carotenoids content by immunohistochemistry, carotenoid binding assay and RNAi. Results showed that the StAR-like-3 encodes a 54.7 kDa transmembrane protein (named as StAR3) of 481 amino acids containing a MENTAL domain and a START (Steroidogenic acute regulatory protein-related lipid transfer) domain, and its expression level in hemocytes and intestine of golden scallops were significantly higher than those of brown ones. Subsequently, the StAR3 protein was detected in the intestinal epithelial cells of golden scallops, and recombinant StAR3 could bind lutein conjugated to protein G and antibody to form a yellow complex, suggesting it is a carotenoid binding protein involving in carotenoids accumulation in golden scallops. Furthermore, total carotenoids content of hemolymph in golden scallops was significantly decreased when the expression of StAR-like-3 suppressed, suggesting this gene plays an important role in transport of carotenoids. Conclusion, the present results indicated that the StAR-like-3 is a key gene responsible for the carotenoids accumulation in the scallop.

Intraspecific hybridization as a mitigation strategy of ocean acidification in marine bivalve noble scallop Chlamys nobilis.

The driving factors of climate change, especially ocean acidification (OA), have many detrimental impacts on marine bivalves. Hybridization is one of the important methods to improve environmental tolerance of animals and plants. In this study, we explored the feasibility of intraspecific hybridization as an OA mitigation strategy in noble scallop Chlamys nobilis (ecologically and economically important bivalve species). The results of this study revealed that exposure of C. nobilis to OA condition significantly reduced the hatching rate, survival rate, growth rate (shell height, shell length, shell width and shell weight), and total carotenoid content (TCC), as well as increased the deformity rate of C. nobilis larvae. Interestingly, under both ambient water and OA condition, the intraspecific hybridization of C. nobilis exhibited heterosis in terms of hatching rate, survival rate and growth rate (excepted for growth in shell length under OA). Transcriptome sequencing of C. nobilis (inbreed and hybrid under ambient and OA conditions) identified four main differentially expressed genes involved in signal transduction, biological process maintenances, nucleic acid binding and post-translational modification. In addition, the expression of these four genes in hybrid C. nobilis was significantly higher than that in inbreed C. nobilis. In conclusion, hybrid C. nobilis showed heterosis in growth rate and survival rate under both ambient water and acidified seawater condition, which may be the result of enhanced expression of genes related to signal transduction, DNA replication and post-translational modification.

Whole-transcriptome RNA sequencing revealed the roles of chitin-related genes in the eyestalk abnormality of a novel mud crab hybrid (Scylla serrata ♀ × S. paramamosain ♂).

Chitin is a kind of insoluble structural polysaccharide and plays different roles in different species. In crustaceans, it forms the structural components in the exoskeleton. In our previous studies, novel mud crab hybrids have been produced from the interspecific hybridization of Scylla serrata ♀ × S. paramamosain ♂. Some of the hybrid crabs have been found to be morphologically (eyestalk) abnormal, but the genetic mechanism remains unknown. To address this question, we performed whole-transcriptome RNA sequencing on the control group (normal hybrids), abnormal hybrids, and S. paramamosain to uncover the genetic basis underlying this morphological abnormality. A total of 695 mRNAs, 10 miRNAs, 44 circRNAs, and 1957 lncRNAs were differentially expressed between normal and abnormal hybrids. Several differentially expressed genes (DEGs) associated with chitin and cuticle metabolism were identified, including chitin synthase, chitinase, chitin deacetylase, ß-N-acetylglucosaminidase, ß-1,4-endoglucanase, N-alpha-acetyltransferase, cuticle proprotein, early cuticle protein, and arthrodial cuticle protein. Functional analysis showed that DE miRNAs, DE circRNAs, DE lncRNAs, and lncRNA/circRNA-miRNA-mRNA network were enriched in pathways related to the amino acid, carbohydrate, and glycogen metabolism. Considering the importance of the chitin and cuticle in exoskeleton formation, it can be concluded that the changes in the chitin and cuticle biosynthesis might have caused the eyestalk abnormality in hybrid crabs. These findings can lay the solid foundation for a better understanding of the important roles of chitin and cuticle related genes and the development of hybridization techniques in crustaceans.

Climate change and n-3 LC-PUFA availability.

Omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) are essential fatty acids for the growth, development and survival of virtually all organisms. There is increasing evidence that anthropogenic climate change has a direct and indirect impact on the availability of natural n-3 LC-PUFA. However, this information is fragmented and not well organized. Therefore, this article reviewed published data from laboratory experiments, field experiments and model simulations to reveal the impact of climate change on the global supply of natural n-3 LC-PUFA and how this will limit the availability of n-3 LC-PUFA in the future food web. In general, climate change can significantly reduce the availability of natural n-3 LC-PUFA in grazing food webs in the following ways: 1) decrease the total biomass of phytoplankton and shift the plankton community structure to a smaller size, which also reduce the biomass of animals in higher trophics; 2) reduce the n-3 LC-PUFA content and/or quality (n-3: n-6 ratio) of all marine organisms; 3) reduce the transfer efficiency of n-3 LC-PUFA in grazing food web. In addition, as an anthropogenic climate adaptation measure, this review also proposed some alternative sources of n-3 LC-PUFA and determined the direction of future research. The information in this article is very useful for providing a critical analysis of the impact of climate change on the supply of natural n-3 LC-PUFA. Such information will aid to establish climate adaptation or management measures, and determine the direction of future research.

Lipid nutritional quality of marine and freshwater bivalves and their aquaculture potential.

Omega-3 Long-chain polyunsaturated fatty acids (n-3 LC-PUFA) are beneficial to human health. Since the industrial revolution, with the tremendous increase of human population, the supply of natural n-3 LC-PUFA is far lower than the nutritional need of n-3 LC-PUFA. Therefore, a new alternative source of natural n-3 LC-PUFA is urgently needed to reduce the supply and demand gap of n-3 LC-PUFA. Mollusks, mainly bivalves, are rich in n-3 LC-PUFA, but the information of bivalves' lipid profile is not well organized. Therefore, this study aims to analyze the published fatty acid profiles of bivalves and reveal the potential of bivalve aquaculture in meeting the nutritional needs of human for n-3 LC-PUFA. There are growing evidence show that the nutritional quality of bivalve lipid is not only species-specific, but also geographical specific. To date, bivalve aquaculture has not been evenly practiced across the globe. It can be seen that aquaculture is predominant in Asia, especially China. Unlike fish aquaculture, bivalve aquaculture does not rely on fishmeal and fish oil inputs, so it has better room for expansion. In order to unleash the full potential of bivalve aquaculture, there are some challenges need to be addressed, including recurrent mass mortalities of farmed bivalves, food safety and food security issues. The information of this article is very useful to provide an overview of lipid nutritional quality of bivalves, and reveal the potential of bivalve aquaculture in meeting the growing demand of human for n-3 LC-PUFA.

Endogenous LC-PUFA biosynthesis capability in commercially important mollusks.

Mollusks are excellent dietary sources for LC-PUFA. However, the main challenge limiting mollusk production is the high mortality rate of molluskan larvae in early life cycle stages. This paper reviews scientific evidences on molecular and biochemical studies of LC-PUFA biosynthesis in commercially important molluskan species. It carefully summarizes the pertinent data published on specific research questions to improve the understanding of the diverse evidences. It is helpful to clarify the current state of research and determine topics for future studies on LC-PUFA biosynthesis in mollusks. From the analysis of published data, mollusks have the ability to biosynthesis LC-PUFA to a certain extent. LC-PUFA biosynthesis information of commercially important molluskan species can be useful to determine the fatty acids essential for their diet. Therefore, specific management strategies or feeds can be developed to strengthen the industry by improving the health and survival rate of molluskan larvae.

Evidence of Sex Differentiation Based on Morphological Traits During the Early Development Stage of Mud Crab Scylla paramamosain.

In order to uncover the sexual difference in morphology and how early they appear during the development stage of mud crab Scylla paramamosain, we measured, observed, and biostatistically analyzed morphological traits related to sex. For unveiling the morphological differences between sexes, morphological traits involving abdomen width (AW), carapace length (CL), and carapace width (CW) were first measured during the crablet development stage of S. paramamosain in the present study. The correlation analyses and path analyses exhibited that sexual dimorphism in the third abdomen width (AW3) and fourth abdomen width (AW4) could be used for sex identification from stage C VI (stage VI of crablet). Based on the stepwise discriminant analysis and standardized traits, a sex discriminant equation was constructed, which is capable for sex identification in crablets from stage C VI. Observations for secondary sexual traits and abdomen morphology (shape and pleopods) using a dissecting microscope or scanning electron microscope indicated that sexes are easily identified at stage C VIII according to the abdomen shape; meanwhile, at stage C II based on pleopod difference, and at stage C I by the presence or absence of gonopores. The findings in this study contribute greatly to the accuracy of sex identification of S. paramamosain during the early development stage, which promotes the understanding of the morphological differentiation mechanism of sex.

Potential causative factors of noble scallop Chlamys nobilis mass mortality in Nan'ao Island, Shantou, China in 2017.

Noble scallop Chlamys nobilis is an important marine bivalve that has been extensively cultured in the south coast of China since the 1980s. Unfortunately, since the late 1990s, the farmed scallops often suffered from regional mass mortality, which results in enormous economic losses to farmers and industries. In 2017, another mass mortality event occurred in Nan'ao Island, Shantou, China. In this study, the cause of C. nobilis mass mortality in 2017 was first investigated in the field, and then validated in a laboratory experiment. In the field, three sampling sites were selected according to the scallop mortality rate: Hunter Bay (90% mortality), Baisha Bay (67% mortality) and Longhai (6% mortality). Meanwhile, environmental parameters (temperature, salinity, DO, pH and chlorophyll a) of each site were also measured in situ. Then, water and scallop samples were collected randomly for the analysis of phytoplankton diversity and algal toxin activity using 18S rDNA and PP2A inhibition assay, respectively. In laboratory, healthy scallops were challenged with Karenia mikimotoi (1 × 103 cells/mL) for 30 h. The field results showed that no significant difference in those environmental parameters existed among the three sites, but the relative abundance of K. mikimotoi in seawater and scallops' intestines in Hunter Bay and Baisha Bay was significantly higher than that in Longhai, and sick scallops contained significantly higher algal toxin activity than healthy ones. Laboratory results revealed that challenged scallops with K. mikimotoi showed significantly higher mortality rate and algal toxin activity than healthy ones, and low density of K. mikimotoi (1 × 103 cells/mL) was sufficient to cause >50% scallops' mortality within 26 h. This study provides the first evidence that low K. mikimotoi cell density can cause massive mortality in C. nobilis, and provides useful information as guide to prevent scallop mass mortality in the future.

mRNA profile provides novel insights into stress adaptation in mud crab megalopa, Scylla paramamosain after salinity stress.

BACKGROUND: Mud crab, Scylla paramamosain, a euryhaline crustacean species, mainly inhabits the Indo-Western Pacific region. Wild mud crab spawn in high-salt condition and the salinity reduced with the growth of the hatching larvae. When the larvae grow up to megalopa, they migrate back to estuaries and coasts in virtue of the flood tide, settle and recruit adult habitats and metamorphose into the crablet stage. Adult crab can even survive in a wide salinity of 0-35 ppt. To investigate the mRNA profile after salinity stress, S. paramamosain megalopa were exposed to different salinity seawater (low, 14 ppt; control, 25 ppt; high, 39 ppt). RESULTS: Firstly, from the expression profiles of Na+/K+/2Cl- cotransporter, chloride channel protein 2, and ABC transporter, it turned out that the 24 h might be the most influenced duration in the short-term stress. We collected megalopa under different salinity for 24 h and then submitted to mRNA profiling. Totally, 57.87 Gb Clean Data were obtained. The comparative genomic analysis detected 342 differentially expressed genes (DEGs). The most significantly DEGs include gamma-butyrobetaine dioxygenase-like, facilitated trehalose transporter Tret1, sodium/potassium-transporting ATPase subunit alpha, rhodanese 1-like protein, etc. And the significantly enriched pathways were lysine degradation, choline metabolism in cancer, phospholipase D signaling pathway, Fc gamma R-mediated phagocytosis, and sphingolipid signaling pathway. The results indicate that in the short-term salinity stress, the megalopa might regulate some mechanism such as metabolism, immunity responses, osmoregulation to adapt to the alteration of the environment. CONCLUSIONS: This study represents the first genome-wide transcriptome analysis of S. paramamosain megalopa for studying its stress adaption mechanisms under different salinity. The results reveal numbers of genes modified by salinity stress and some important pathways, which will provide valuable resources for discovering the molecular basis of salinity stress adaptation of S. paramamosain larvae and further boost the understanding of the potential molecular mechanisms of salinity stress adaptation for crustacean species.

Differential responses of a pi-class glutathione S-transferase (CnGSTp) expression and antioxidant status between golden and brown noble scallops under pathogenic stress.

Glutathione S-transferases (GSTs) play important roles in immunity by protecting organisms against the damage of reactive oxygen species (ROS). In this study, a pi-class GST cDNA sequence was first cloned from noble scallop Chlamys nobilis (named CnGSTp). The full length cDNA of CnGSTp was 922 bp, encoding a cytosolic protein of 202 amino acids residues, with predicted molecular masses of 23.1 kDa. Then an acute Vibrio Parahaemolyticus challenge experiment was conducted by using the Golden and Brown noble scallops with different total carotenoids content (TCC), and CnGSTp expression level, TCC and ROS level was separately determined. The results showed that ROS and CnGSTp expression levels were significantly up-regulate under Vibrio Parahaemolyticus challenge than the control group (P < 0.05). The Golden scallops showed significantly higher CnGSTp expression level and lower ROS level in hemocytes than the Brown ones (P < 0.05). Moreover, there is a significantly positive correlation between TCC and ROS in the Golden scallops. The present results revealed that CnGSTp plays important roles in immune response and carotenoids play assistant roles in antioxidant defense system under pathogenic stress in the noble scallop.

Effects of high stocking density on the galectin gene expression in noble scallop Chalmys nobilis under bacterial infection.

High stocking densities have been shown to have adverse effects on the physiology of bivalves. The noble scallop Chlamys nobilis is one of the most important cultured shellfish in Southern China. However, the effects of scallop stocking density on its immunity is not well understood. In this context, this study was conducted to assess the effect of high stocking density on the galectin (an important protein in innate immunity) gene expression of C. nobilis during bacterial infection. A full-length galectin (CnGal) gene was cloned. The ORF of the CnGal cDNA encodes a predicted protein containing 549 aa with four CRDs and no signal peptide. Our results reveal that high stocking density in the scallop not only led to high mortality and slow growth, but also changed tissue distribution of the CnGal expression. The individuals from the high stocking density group exhibited more differences among tissues than those from the control group, but the highest expression were both recorded in hemolymph. After the Vibrio parahaeomlyticus challenge, the gene's expression levels were all significantly up-regulated in the hemolymph and gill, but the time up to peak was different between the two tissues. The findings of this study could fill a gap in knowledge about how high stocking density affect scallop immunity at the molecular level.

Identification and molecular characterization of peroxiredoxin 6 from noble scallop Chlamys nobilis revealing its potent immune response and antioxidant property.

The 1-cyseine peroxiredoxin (Prx6) is an importantly antioxidant enzyme that protects cells from oxidative damage caused by excessive production of reactive oxygen species (ROS). In this study, we described the molecular characteristics of the noble scallop Chlamys nobilis peroxiredoxin 6 (designed as CnPrx6), immune responses and DNA protection activity of the recombinant protein. The complete ORF (696 bp) of CnPrx6 encoded a polypeptide (25.5 kDa) of 231 amino acids, harboring a conserved peroxidase catalytic center (41PVCTTE46) and the catalytic triads putatively involved in peroxidase and phospholipase A2 activities. The deduced amino acid sequence of CnPrx6 shared a relatively high amino acid sequence similarity (more than 50%). The qRT-PCR revealed that the CnPrx6 mRNA was constitutively expressed in all examined tissues, with the highest expression observed in adductor. Upon immunological challenge with Vibrio parahaemolyticus, lipopolysaccharides (LPS) and polyinosinic-polycytidylic acid (Poly I:C), the expression level of CnPrx6 mRNA was significantly up-regulated (P < 0.05). Furthermore, there was a significant difference (P < 0.05) in the expression level of CnPrx6 between golden and brown scallops. The purified recombinant CnPrx6 protein protected the supercoiled plasmid DNA from metal-catalyzed ROS damage. Taken together, these results indicated that the CnPrx6 may play an important role in modulating immune responses and minimizing DNA damage in noble scallop Chlamys nobilis.

A novel carotenoids-producing marine bacterium from noble scallop Chlamys nobilis and antioxidant activities of its carotenoid compositions.

Marine bacteria produce many bioactive compounds, including carotenoids. However, the quality of bacterium carotenoids is relatively unknown. Therefore, in this study, a novel carotenoids-producing bacterium Brevundimonas scallop Zheng & Liu was isolated from Chlamys nobilis. The genome of the isolate was analyzed, carotenoid compounds were screened using HPLC-MS and the carotenoid production in B. scallop was monitored. The results revealed that the genome of B. scallop contained a carotenoid synthesis gene cluster, which involved in astaxanthin and hydroxy-astaxanthin biosynthesis. The 2,2'-dihydroxy-astaxanthin was the major carotenoid produced by B. scallop. The optimum culture condition for the highest carotenoids production (1303.62 ± 61.06 µg/g dry cells) for B. scallop was at temperature and salinity of 20 °C and 3% salt, respectively, in 10 g/L glucose as carbon source. The results showed the B. scallop is a new carotenoids resource in marine bivalve, which has an excellent antioxidative activity and potential industrial use.