Variations in retention efficiency of bivalves to different concentrations and organic content of suspended particles

Retention efficiencies (RE) of scallop (Chlamys farreri), oyster (Crassostrea gigas) and mussel (Mytilus edulis) in a flow-through system were measured to understand the short-term response to various particle and organic matter concentrations. By comparing the RE of C. farreri with that of C. gigas and M. edulis, we gained further knowledge on the feeding physiological characteristics of C. farreri and ascertained the possible cause of high summer mortalities of this species. The experimental feeding conditions included natural differences in the abundance and composition of suspended seston, as well as conditions in which seston abundance and composition were manipulated by adding natural silt or cultured microalgae. The results show that in natural sea water, the minimum particle size for maximal retention in M. edulis, C. gigas and C. farreri was approximately 4, 6, and 8 μm, respectively; the RE of 2-μm (equivalent spherical diameter) particles was 17%, 19%, and 8%, respectively; and the relative standardized RE was 58%, 49%, and 18%, respectively. In C. gigas and M. edulis, the minimal particle size for maximal retention did not change with food quality (organic content). C. farreri was more sensitive to fluctuations in particle concentration and organic content. With particle concentration increase, the minimal particle size for maximal retention in scallop shifted to large particles and the RE for 2-μm particle decreased from 8% in natural seawater to 1.6%–6% in silt-enriched groups. With organic content increase, the minimal particle size for maximal retention shifted from 8 to 5 μm in natural seawater.Variation in RE of C. farreri with food conditions and the relationship between lower RE and smaller particle size may hamper C. farreri from food taking due to the decrease in the size distribution of phytoplankton in Sungo Bay.     

Iodothyronine deiodinase gene analysis of the Pacific oyster Crassostrea gigas reveals possible conservation of thyroid hormone feedback regulation mechanism in mollusks

Iodothyronine deiodinase catalyzes the initiation and termination of thyroid hormones(THs) effects, and plays a central role in the regulation of thyroid hormone level in vertebrates. In non-chordate invertebrates, only one deiodinase has been identified in the scallop C hlamys farreri. Here, two deiodinases were cloned in the Pacific oyster C rassostrea gigas( Cg Dx and C g Dy). The characteristic in-frame TGA codons and selenocysteine insertion sequence elements in the oyster deiodinase c DNAs supported the activity of them. Furthermore, seven orthologs of deiodinases were found by a tblastn search in the mollusk Lottia gigantea and the annelid C apitella teleta. A phylogenetic analysis revealed that the deiodinase gene originated from an common ancestor and a clade-specific gene duplication occurred independently during the differentiation of the mollusk, annelid, and vertebrate lineages. The distinct spatiotemporal expression patterns implied functional divergence of the two deiodinases. The expression of C g Dx and Cg Dy was influenced by L-thyroxine T4, and putative thyroid hormone responsive elements were found in their promoters, which suggested that the oyster deiodinases were feedback regulated by TH. Epinephrine stimulated the expression level of C g Dx and Cg Dy, suggesting an interaction effect between different hormones. This study provides the first evidence for the existence of a conserved TH feedback regulation mechanism in mollusks, providing insights into TH evolution.     

Morphological beak differences of loliginid squid,Uroteuthis chinensis and Uroteuthis edulis,in the northern South China Sea

The mitre squid(U roteuthis chinensis) and the swordtip squid( U. edulis) are Indo-Pacific cephalopod species that are abundant in the western Pacific Ocean. They are currently exploited in the East and South China Seas and make up a significant portion of the Chinese neritic squid catch. Beaks, the feeding organs of squid, are important for individual size and biomass estimation because of their high resistance to degradation in predator stomachs and consistent dimensions. In this study, 104 U. chinensis and 143 U. edulis individuals were sampled from northern South China Sea with mantle length from 70 to 260 mm and 96 to 284 mm, respectively. The results indicated that morphological beak values were greater for U. edulis, compared to U. chinensis, for upper hood length(UHL), upper crest length(UCL), upper lateral wall length(ULWL), lower crest length(LCL), and lower lateral wall length(LLWL). According to principal component analysis, UHL/ML, UCL/ML, ULWL/ML, LCL/ML, LLWL/ML and LWL/ML could represent the characteristics of beaks for U. chinensis, while UHL/ML, UCL/ML, ULWL/ML, LHL/ML, LCL/ML and LLWL/ML could represent it for U. edulis. According to Akaike's information criterion(AIC) values, a power function was the most suitable model for U. chinensis, while a linear function was the most suitable model for U. edulis. The beak variable-mantle length ratio(beak variable/mantle length) declined with the increasing of mantle length and declined sharply at the early stage of growth in both beaks and species. The ratio changed quickly after achieving the mantle length of 140 mm for U. chinensis, while the ratio changed quickly after 170 mm for U. edulis. Beaks in both species experienced sharper changes through maturity stage I to II than other maturity stages. This study gives us basic beak morphology information for U. chinensis and U. edulis in the East and South China Seas. Geometric morphological methods combined with dietary analysis should be used in the future.     

Population dynamics of four dominant copepods in Prydz Bay, Antarctica, during austral summer from 1999 to 2006

Population dynamics of four dominant Antarctic copepods, Calanoides acutus, Calanus propinquus, Metridia gerlachei and Rhincalanus gigas were studied based on zooplankton samples collected in the Prydz Bay during austral summer from 1999 to 2006. We found that C. acutus was the most abundant species among these four copepods, followed by C. propinquus, M. gerlachei and R. gigas. R. gigas occurred mainly in the warmer oceanic regions and showed distribution patterns discrete from the other three species, whose distribution in the whole survey area overlapped. By December 15th (about one month before our sampling) of the years 1999, 2003 and 2006, sea ice retreated earlier and polynyas existed in the neritic region one month before sampling. These periods were characterized by numerical dominance of C. acutus, C. propinquus and M. gerlachei, elevated proportions of Copepodite I and Copepodite II stages especially in the neritic region. While for the years 2000, 2002, and 2005, the ice edge located more northerly and polynyas did not exist in the neritic region, the copepods abundance was lower, indicating poor recruitment. Population structure of R. gigas was mainly composed of advanced stages Copepodite V and female during all cruises. Log10 (x+1) transformed densities of C. acutus, C. propinquus and M. gerlachei showed positive correlation with temperature and chlorophyll a concentration, while mean population stages of these copopods were negatively correlated with these environmental variables. Younger copepodite stages of C. acutus, C. propinquus and M. gerlachei appeared more often in neritic regions. We confirmed that the polynyas had a great contribution to phytoplankton blooms, which promote copepods reproduction and recruitment success. The study suggested that population dynamics of the four copepods have good correspondence with sea ice and polynya variations during all cruises of the Prydz Bay.     

CYTOLOGICAL COMPARISON OF GAMETOGENESIS OF SCALLOPS,ARGOPECTEN IRRADIANS AND CHLAMYS FARRERI

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Relationship Between Single Nucleotide Polymorphism of Glycogen Synthase Gene of Pacific Oyster Crassostrea gigas and Its Glycogen Content

Glycogen is important not only for the energy supplementary of oysters, but also for human consumption. High glycogen content can improve the stress survival of oyster. A key enzyme in glycogenesis is glycogen synthase that is encoded by glycogen synthase gene GYS. In this study, the relationship between single nucleotide polymorphisms(SNPs) in coding regions of Crassostrea gigas GYS(Cg-GYS) and individual glycogen content was investigated with 321 individuals from five full-sib families. Single-strand conformation polymorphism(SSCP) procedure was combined with sequencing to confirm individual SNP genotypes of Cg-GYS. Least-square analysis of variance was performed to assess the relationship of variation in glycogen content of C. gigas with single SNP genotype and SNP haplotype. As a consequence, six SNPs were found in coding regions to be significantly associated with glycogen content(P 0.01), from which we constructed four main haplotypes due to linkage disequilibrium. Furthermore, the most effective haplotype H2(GAGGAT) had extremely significant relationship with high glycogen content(P 0.0001). These findings revealed the potential influence of Cg-GYS polymorphism on the glycogen content and provided molecular biological information for the selective breeding of good quality traits of C. gigas.     

Development of Genomic Microsatellite Multiplex PCR Using Dye-Labeled Universal Primer and Its Validation in Pedigree Analysis of Pacific Oyster(Crassostrea gigas)

There is an increasing requirement for traceability of aquaculture products, both for consumer protection and for food safety. There are high error rates in the conventional traceability systems depending on physical labels. Genetic traceability technique depending on DNA-based tracking system can overcome this problem. Genealogy information is essential for genetic traceability, and microsatellite DNA marker is a good choice for pedigree analysis. As increasing genotyping throughput of microsatellites, microsatellite multiplex PCR has become a fast and cost-effective technique. As a commercially important cultured aquatic species, Pacific oyster Crassostrea gigas has the highest global production. The objective of this study was to develop microsatellite multiplex PCR panels with dye-labeled universal primer for pedigree analysis in C. gigas, and these multiplex PCRs were validated using 12 full-sib families with known pedigrees. Here we developed six informative multiplex PCRs using 18 genomic microsatellites in C. gigas. Each multiplex panel contained a single universal primer M13(-21) used as a tail on each locus-specific forward primer and a single universal primer M13(-21) labeled with fluorophores. The polymorphisms of the markers were moderate, with an average of 10.3 alleles per locus and average polymorphic information content of 0.740. The observed heterozygosity per locus ranged from 0.492 to 0.822. Cervus simulations revealed that the six panels would still be of great value when massive families were analysed. Pedigree analysis of real offspring demonstrated that 100% of the offspring were unambiguously allocated to their parents when two multiplex PCRs were used. The six sets of multiplex PCRs can be an important tool for tracing cultured individuals, population genetic analysis, and selective breeding program in C. gigas.     

Isozyme expression of Chinese and Japanese populations of Chlamys farreri and their reciprocal hybrids

Chinese and Japanese population of Chlamys farreri and their reciprocal hybrids were surveyed in isozyme variability at 13 loci by polyacrytamide gel electrophoresis (PAGE). Isozyme banding patterns indicated these hybrids were diploid. Loci that were observed as being monomorphic in inbred populations of C. farreri were also found to be monomorphic in filial progeny; loci that observed to be polymorphic in parental type generations were also polymorphic in hybrid generations. Differences existed among allelic frequency of the four types of cross. Within the reciprocal hybrids the expression of malic enzyme (ME) isozyme was sufficient to distinguishing individual hybrids because of the band, Rf=0.38. However, there were no noticeable variations among all the samples to differentiate one from another. Inbreeding was likely to be the main problem in aquaculture. The introduction of new broodstock can improve the genetic diversity. Hybrid vigor has manifested to a certain extent in the present study.     

Polypeptide from Chlamys farreri inhibits UVB-induced apoptosis of HaCaT cells via iNOS/NO and HSP90

Polypeptide from Chlamys farreri(PCF) is a novel marine bioactive product that was isolated from the gonochoric Chinese scallop Chlamys farreri,and was found to be an effective antioxidant in our recent studies.In this study,we investigated the effect of PCF on ultraviolet B(UVB)-induced apoptosis of HaCaT cells and the intracellular signaling pathways involved.Pretreatment with the inducible nitric oxide synthase(iNOS) inhibitor S-methylisothiourea sulfate inhibited UVB-induced apoptosis,indicating that iN...     

Cytochemical characterization of yolk granule acid phosphatase during early development of the oyster Crassostrea gigas (Thunberg)

In this study,a cytochemical method and transmission electron microscopy was used to examine acid phosphatase activities of yolk granules throughout the early developmental stages of the Pacific oyster Crassostrea gigas. This study aimed to investigate the dynamic change of yolk granule acid phosphatase,and the mechanisms underlying its involvement in yolk degradation during the early developmental stages of molluscs. Three types of yolk granules(YGI,YGII,and YGIII) that differed in electron density and acid phosphatase reaction were identified in early cleavage,morula,blastula,gastrula,trochophore,and veliger stages. The morphological heterogeneities of the yolk granules were related to acid phosphatase activity and degrees of yolk degradation,indicating the association of acid phosphatase with yolk degradation in embryos and larvae of molluscs. Fusion of yolk granules was observed during embryogenesis and larval development of C. gigas. The fusion of YGI(free of acid phosphatase reaction) with YGII(rich in acid phosphatase reaction) could be the way by which yolk degradation is triggered.     

Relationship Between Shell Color and Growth and Survival Traits in the Pacific Oyster Crassostrea gigas

Shell color is regarded as an economic trait in many breeding programs of bivalve mollusks, but the relationship between shell color and production traits remains controversial. In our breeding program of Crassostrea gigas, solid white, black, and orange shell lines were obtained, and second-generation(F2) and testcross families with segregating shell colors were constructed by crossing these three lines. These segregating families provided ideal samples for investigating the relationship between shell color and production traits in C. gigas. The growth and survival of 7-month-old oysters with different shell colors sampled within the same families were compared in seven F2 families and 13 testcrosses. In addition, the growth and survival of oysters from the three shell color lines were compared at 4 and 16 months of age. The growth and survival rates of the orange shell line were significantly lower than those of the white and black shell lines. However, no significant difference in growth between oysters with different shell colors was observed within segregating families, except the testcrosses produced by crossing orange-and white-shelled grandparents, and no significant difference in survival was observed in any family. Overall, no significant correlation was observed between shell color and production traits in C. gigas. These results suggest that shell color cannot be used as a marker to guide the selection of growth and survival traits. Thus shell color and production traits should be selected independently in oyster breeding programs.     

Impacts of El Ni?o on the somatic condition of Humboldt squid based on the beak morphology

The Humboldt squid Dosidicus gigas has a short life span,and environmental variability plays a significant role in regulating its population dynamics and distribution.An analysis of 1 096 samples of D.gigas collected by the Chinese commercial fishing vessels during 2013,2014,and 2016 off the Peruvian Exclusive Economic Zone,was conducted to evaluate the impacts of El Ni?o events on the somatic condition of D.gigas.This study indicates that the slopes of all beak variables in relation to mantle length(ML)for females were greater than those of males during 2013,2014,and 2016,and slopes of the upper crest length and the lower ro strum length significantly differed between females and males in 2013(P0.05).Variation in the slopes for beak variables among years was studied;no significant difference was observed(ANCOVA,P0.05).The Fulton's condition coefficients(K)of females and males in 2013 and 2014 were significantly greater than tho se in 2016(P0.01).01).The K values of females were greater than tho se of males in 201 3,2014,and 2016,and K values significantly differed between females and males in 201 3.In normal years,the chlorophyll a(Chl a)concentration showed an N-shaped variability from January to December.However,in the El Ni?o period,it tended to weaken the upwelling coupled with warm and low Chl a concentration waters.We suggest that the poor somatic condition of D.gigas during the El Ni?o year was resulted from the low Chl a concentration in the waters,and the abundance of D.gigas would decrease due to the unfavourable environment and the lack of prey items in the El Ni?o year.     

Protective Effect of Topically Applied Polypeptide from Chlamys farreri Against Ultraviolet Radiation-Induced Chronic Skin Damage in Guinea Pig

INTRODUCTIONSolarultraviolet(UV)radiationisknowntobeamajoretiologicalriskofphotoaging.AsUVC(2 0 0nm -2 90nm)isabsorbedentirelybythestratosphericozonelayer,sunlightontheearthsur faceiscomposedofUVA (3 2 0nm -4 0 0nm)andUVB (2 90nm -3 2 0nm)only.UVAradiationenergyaccountsformorethan 90 %ofsolarUV ;andpenetratesthebasallayerofepidermismoreefficientlythanUVB (Brulsetal.,1984) .However,atsimilardosages,UVBismuchmoremuta genicthanUVA (Drobetskyetal.,1995) .Furthermore ,humanskinisexp…     

Molecular Identification of Dried Shellfish Products Sold on the Market Using DNA Barcoding

The dried shellfish products with rich nutrients and low-calorie content are favorite food in China, especially in coastal areas. However, the species of dried shellfish products in the market are usually unknown, as the taxonomic features were removed during the production process. This study described the application of DNA barcoding technique to the identification of 100 dried shellfish(scallop, squid, octopus and cuttlefish) products in markets. Samples were authenticated by comparing mitochondrial cytochrome oxidase subunit I(COI) gene and 16 S ribosomal RNA(16 S rRNA) gene sequences with public reference taxonomic databases. The results showed that all the 100 products can be identified at species level. Sixty four scallop adductor products were processed using the bay scallop, Argopecten irradians, and one was from Portuguese oyster, Crassostrea angulata. All the 27 squid, 2 cuttlefish and 6 octopus products were produced by the Jumbo flying squid, Dosidicus gigas. The neighbour-joining tree is in agreement with the results of DNA barcoding analysis. The 64 scallop samples formed one A. irradians cluster, leaving Sca65 clustered with the reference oyster sequence C. angulata(MH997922). All the 35 cephalopod(squid, octopus and cuttlefish) samples formed a D. gigas cluster. This investigation revealed a low variety of dried shellfish products sold on the market, and highlighted the high rate of mislabeling and species substitution. Our present work provides a practical method for tracing and authenticating shellfish products.     

Transcriptome Analysis of Crassostrea sikamea (♀) × Crassostrea gigas (♂) Hybrids Under and After Thermal Stress

Crossbreeding is an effective approach to manage the genetic decline in aquaculture.One-way hybrids of Crassostrea sikamea(♀)and Crassostrea gigas(♂)have advantages in growth traits and adaptation to high temperature.Here,we used high-throughput sequencing to analyze the molecular processes in the hybrids under and after thermal stress.The hybrids were cultured in the seawater with an increasing temperature from 25℃to 40℃during 10 hours,which is regarded as the thermal stress stage.Then the temperature decreased from 40℃to 25℃within 2 h,which is regarded as the recovery stage.In this study,1293 significant diffe-rentially expressed genes(DEGs)were obtained under thermal stress,of which 576 were upregulated and 717 were downregulated,and 740 significant differentially expressed genes(DEGs)were obtained in the recovery stage,of which the number of upregulated and downregulated genes was 409 and 331,respectively.The antigen processing and presentation,NOD-like,and NF-kappa B path-ways were significantly enriched during the thermal stress stage.The MAPK and PPAR signaling pathways were significantly enrich-ed during the recovery stage.The HSP70,HSP90,and CANX genes were strongly and rapidly upregulated in the control/thermal stress groups but were slightly less upregulated in the thermal stress/recovery group.These results indicate that the innate immune system or nonspecific immunity was deployed to protect interior tissues from thermal stress.In addition,85%of the mutual DEGs were involved in bidirectional regulation(up/down or down/up)when the oysters were removed from the thermal stress to recover.This study provides preliminary insight into the molecular response of C.sikamea(♀)and C.gigas(♂)hybrids to thermal stress and provides a basis for future studies on temperature-adaptation and the possible expansion of hybrid breeding.