Species composition and distribution of common Crassostrea and Saccostrea oysters along the coast of Hainan Island

Oyster cultivation is a pivotal economic industry in Hainan Island,South China,where a high oyster species diversity is shown.However,the specific distribution and biodiversity of oyster resources in the island have remained unclear.To elucidate the diversity of oyster species and their distribution there,307 oyster samples were collected from 29 sites in the intertidal zone around the island,and were identified using both morphological and molecular approaches.A minimum of 12 oyster species were identified in taxonomy,including Crassostrea species(C.gigas angulata,C.sikamea,C.iredalei,C.dianbaiensis,C.talonata,C.ariakensis,and C.hongkongensis),and Saccostrea species(S.malabonensis,S.mordax,S.echinata,S.circumsuta,and S.mordoides).The results revealed a remarkable diversity of oyster species along the coast of the island.Particularly noteworthy are that S.malabonensis and S.mordax constituted 36%and 22%of the collected specimens,respectively.This study provided a comprehensive overview on current state of oyster biodiversity in Hainan,serving as a valuable reference for conservation and research on species distribution and resource dynamics in oyster populations.

Three novel umami peptides derived from the alcohol extract of the Pacific oyster(Crassostrea gigas):identification,characterizations and interactions with T1R1/T1R3 taste receptors

Oyster(Crassostrea gigas),the main ingredient of oyster sauce,has a strong umami taste.In this study,three potential umami peptides,FLNQDEEAR(FR-9),FNKEE(FE-5),and EEFLK(EK-5),were identified and screened from the alcoholic extracts of the oyster using nano-HPLC-MS/MS analysis,i Umami-Scoring Card Method(i Umami-SCM)database and molecular docking(MD).Sensory evaluation and electronic tongue analysis were further used to confirm their tastes.The threshold of the three peptides ranged from 0.38 to 0.55 mg/m L.MD with umami receptors T1R1/T1R3 indicated that the electrostatic interaction and hydrogen bond interaction were the main forces involved.Besides,the Phe592 and Gln853 of T1R3 were the primary docking site for MD and played an important role in umami intensity.Peptides with two Glu residues at the terminus had stronger umami,especially at the C-terminus.These results contribute to the understanding of umami peptides in oysters and the interaction mechanism between umami peptides and umami receptors.

Spatial compartmentalization and temporal stability of associated microbiota in Pacific oyster Crassostrea gigas

The Pacific oyster Crassostrea gigas,one of the most exploited molluscs in the world,has suffered from massive mortality in recent decades,and the occurrence mechanisms have not been well characterized.In this study,to reveal the relationship of associated microbiota to the fitness of oysters,temporal dynamics of microbiota in the gill,hemolymph,and hepatopancreas of C.gigas during April 2018-January 2019 were investigated by 16 S rRNA gene sequencing.The microbiota in C.gigas exhibited tissue heterogeneity,of which Spirochaetaceae was dominant in the gill and hemolymph while Mycoplasmataceae enriched in the hepatopancreas.Co-occurrence network demonstrated that the gill microbiota exhibited higher inter-taxon connectivity while the hemolymph microbiota had more modules.The richness(Chao 1 index)and diversity(Shannon index)of microbial community in each tissue showed no significant seasonal variations,except for the hepatopancreas having a higher richness in the autumn.Similarly,beta diversity analysis indicated a relatively stable microbiota in each tissue during the sampling period,showing relative abundance of the dominant taxa exhibiting temporal dynamics.Results indicate that the microbial community in C.gigas showed a tissue-specific stability with temporal dynamics in the composition,which might be essential for the tissue functioning and environmental adaption in oysters.This work provides a baseline microbiota in C.gigas and is helpful for the understanding of host-microbiota interaction in oysters.

Non-Targeted Metabolomics Reveals the Metabolic Alterations in Response to Artificial Selective Breeding in the Fast-Growing Strains of Pacific Oyster

Pacific oyster(Crassostrea gigas)is one of the most important mollusks cultured all around the world.Selective breeding programs of Pacific oysters in China is initiated since 2006 and developed the genetically improved strain with fast-growing trait.However,little is known about the metabolic signatures of the fast-growing trait.In the present study,the non-targeted metabolomics was performed to analyze the metabolic signatures of adductor muscle tissue in one-year old Pacific oysters from fast-growing strain and the wild population.A total of 7767 and 10174 valid peaks were extracted and quantified in ESI^(+)and ESI^(−)modes,resulting in 399 and 381 annotated metabolites,respectively.PCA and OPLS-DA revealed that considerable separation among samples from fastgrowing strain and wild population,suggesting the differences in metabolic signatures.Meanwhile,81 significantly different metabolites(SDMs)were identified in the comparisons between fast-growing strain and wild population,based on the strict thresholds.It was found that there were highly correlation and conserved coordination among these SDMs.KEGG enrichment analysis indicated that the SDMs were tightly related to pantothenate and CoA biosynthesis,steroid hormone biosynthesis,riboflavin metabolism,and arginine and proline metabolism.Of them,the CoA biosynthesis and metabolism,affected by pantetheine and pantothenic acid,might be important for the growth of Pacific oysters under artificial selective breeding.The study provides the comprehensive views of metabolic signatures in response to artificially selective breeding,and is helpful to better understand the molecular mechanism of fastgrowing traits in Pacific oysters.

Characterisation of the Bacteria and Archaea Community Associated with Wild Oysters, At Three Possible Restoration Sites in the North Sea

With 85% of the global oyster reefs destroyed, there is an urgent need for large scale restoration to benefit from the ecosystem services provided by biogenic oyster reefs and their associated biodiversity, including microorganisms that drive marine biogeochemical cycles. This experiment established a baseline for the monitoring of the bacterial and archaeal community associated with wild oysters, using samples from their immediate environment of the Voordelta, with cohabiting Crassostrea gigas and Ostrea edulis, Duikplaats with only C. gigas attached to rocks, and the Dansk Skaldyrcentre, with no onsite oysters. The microbial profiling was carried out through DNA analysis of samples collected from the surfaces of oyster shells and their substrate, the sediment and seawater. Following 16S rRNA amplicon sequencing and bioinformatics, alpha indices implied high species abundance and diversity in sediment but low abundance in seawater. As expected, Proteobacteria, Bacteroidetes, Firmicutes and Thaumarchaeota dominated the top 20 OTUs. In the Voordelta, OTUs related to Colwellia, Shewanella and Psychrobium differentiated the oysters collected from a reef with those attached to rocks. Duikplaats were distinct for sulfur-oxidizers Sulfurimonas and sulfate-reducers from the Sva 0081 sediment group. Archaea were found mainly in sediments and the oyster associated microbiome, with greater abundance at the reef site, consisting mostly of Thaumarchaeota from the family Nitrosopumilaceae. The oyster free site displayed archaea in sediments only, and algal bloom indicator microorganisms from the Rhodobacteraceae, Flavobacteriaceae family and genus [Polaribacter] huanghezhanensis, in addition to the ascidian symbiotic partner, Synechococcus. This study suggests site specific microbiome shifts, influenced by the presence of oysters and the type of substrate.

Effect of Fouling on Feeding, Oxygen Consumption and Waste Excretion of Pearl Oyster Pinctada martensii in Daya Bay Cultivation

Biofouling is a particular problem in the pearl oyster culture. It may reduce the growth and survival rates of the cultured species. Foulers are often themselves filter feeders, and are therefore potential competitors for food resource with the cultured species. Fouling organisms may also reduce the oxygen supply. A study was conducted to measure the impact of foulers on feeding oxygen consumption, and waste excretion. POM, ammonia, phosphate and oxygen concentration were measured in various treatments （cultured species, foulers）. This study showed that fouling organisms had significant effect on food uptake, oxygen consumption and waste excretion. The clearance rate, ammonia and phosphate release rate, oxygen consumption rate of the fouled pearl oyster were greater than those of the clean pearl oysters. Other foulers that settled on cages or buoys also contributed much to phytoplankton depletion, oxygen consumption and concentration increase of ammonia and phosphate in water. Therefore, this study showed us that foulers were important competitors in the pearl oyster cultivation of Daya Bay in November 2005.

Heavy metals in oysters, mussels and clams collected from coastal sites along the Pearl River Delta,South China

Concentrations of 8 heavy metals: cadmium (Cd), copper (Cu), zinc (Zn), lead (Pb), nickel (Ni), chromium (Cr), antimony (Sb) and tin (Sn) were examined in 3 species of bivalves (Perna viridis, Crassostrea rivularis and Ruditapes philippinarum) collected from 25 sites along the Pearl River Delta coastal waters in the South China Sea from July to August 1996. In general, Cd, Cu, Zn and Sn concentrations in the three bivalve species collected from the Estuarine Zone were significantly higher than those collected from the Western and Eastern Zones of the Pearl River Delta, which are related to the existence of various anthropogenic activities in the catchment of the Pearl River Delta. The Western Estuarine Zone is mainly impacted by Cr, Ni and Cu contamination. In Victoria Harbor, heavy metal contamination is mainly due to Cu and Pb. Cd, Cu and Zn concentrations in oysters were significantly higher than those in mussels and clams. This could be explained by the fact that oysters live mainly in the Estuarine Zone of the Pearl River Delta which receives most of the polluting discharges from the catchment of the Delta. During turbid condition, heavy metals(soluble or adsorbed on suspended particulates) discharged from the Delta are filtered from the water column and subsequently accumulated into the soft body tissues of oysters. Heavy metal concentrations in the three bivalve species were compared with the maximum permissible levels of heavy metals in seafood regulated by the Public Health and Municipal Services Ordinance, Laws of Hong Kong, and it was revealed that Cd and Cr concentrations in the three bivalve species exceeded the upper limits. At certain hotspots in the Delta, the maximum acceptable daily load for Cd was also exceeded.

Comparative Study on the Antioxidant Activity of Peptides from Pearl Oyster(Pinctada martensii) Mantle Type V Collagen and Tilapia(Oreochromis niloticus) Scale Type Ⅰ Collagen

In this study, Pearl oyster mantle type V collagen(POMC) and tilapia scale type I collagen(TSC) were extracted and hydrolyzed by various proteases in order to obtain peptides. The antioxidant activity of the peptides was investigated by DPPH, hydroxyl radical scavenging experiments and a dynamic digestion model in vitro. The results show that there are significant differences in amino acid composition between POMC and TSC. The collagen peptides obtained from pearl oyster mantle(POMCP) by treating with alkaline protease exhibited higher antioxidant activity than that from tilapia scale(TSCP) treated with papaya protease, and both of them showed greater DPPH and hydroxyl radical scavenging activity than other peptides. After being separated via Sephadex G-25 chromatography, the M1 fraction isolated from POMCP, and the S1 fraction from TSCP with which both had higher molecular weights showed the strongest antioxidant activity than other fractions, and the M1 fraction exhibited stronger antioxidant activity than the S1 fraction in scavenging free-radicals and protecting cells from the oxidation damage. Furthermore, after treating the dynamic digestion system model in vitro, the DPPH and hydroxyl radical scavenging activity of the M1 fraction increased slightly. These results suggest that POMCP exhibits stronger antioxidant activity than TSCP, which means that PMOP may be a good candidate to be a potential natural antioxidant in the food-processing industry.

Preparation, Characteristics, and Formation Mechanism of Oyster Peptide-Zinc Nanoparticles

Oyster peptide zinc nanoparticles (OPZNPs)(28 108 nm) were prepared in the presence of 0.5% 0.9% zinc sulfate at pH 6.0 11.0. The obtained nanoparticles exhibited uniform size distribution and spherical shapes. Nanoparticle characteristics, such as size, surface charge, and hydrophobicity, could be adjusted by controlling zinc sulfate concentration and environmental pH. In- creasing pH value or decreasing zinc sulfate concentration tended to reduce nanoparticle size and increase nanoparticle surface charge and hydrophobicity. OPZNPs presented good stability at near-neutral pH and could be stored for at least 20 days at 4℃. The results of the peptide conformation study and nanoparticle dissociation test proved that zinc ions and carboxyl groups are the key factors that affect OPZNP formation. The intermolecular combinations of carboxyl groups via zinc bridging facilitated the aggrega- tion of oyster peptides. Nanoparticle formation was accompanied by aggregate association and conformational changes. These changes included increments in β-sheets, especially intermolecular β-sheets, at the expense of α-helixes. Overall, this work provided a green alternative route for the synthesis of OPZNPs.

Changes in the Contents of ATP and Its Related Breakdown Compounds in Various Tissues of Oyster During Frozen Storage

The changes in the contents of adenosine 5’-triphosphate (ATP) and its related breakdown compounds were investigated in the adductor muscle, mantle, gill and body trunk of oyster (Crassostrea gigas) during frozen storage at -20℃ and -30℃ and compared with that of the fresh oyster. The investigation was performed using an HPLC system. Different extents of ATP decomposi- tion were found in various tissues frozen at the two temperatures. The K, K’ and A.E.C values were calculated as the chemical fresh- ness indices. Considering the results of sensory evaluation, the A.E.C. value in body trunk at -20℃ and -30℃ could be used as the best freshness index for frozen oyster.

High Pressure Treatment Changes Spoilage Characteristics and Shelf Life of Pacific Oysters(Crassostrea gigas)During Refrigerated Storage

The effects of high pressure(HP) treatment on spoilage characteristic and shelf life extension of Pacific oysters(Crassostrea gigas) during refrigerated storage were studied.Results showed that HP treatment of 275 MPa for 3 min or 300 MPa for 2 min could achieve 100% full release of oyster adductor muscle,pressures higher than 350 MPa caused excessive release as the shells of oysters were broken,thus use of higher pressures should be cautious in oyster processing industry because of its adverse impact on the appearance of shells.HP treatment(300 MPa,2 min) was proper for the shucking of Pacific oyster(Crassostrea gigas) in China.This treatment caused no organoleptic disadvantage.Moreover,HP treatment resulted in obvious differences in biochemical spoilage indicators(p H,TVB-N and TBARS) changes and volatile compounds profile determined by electronic nose during storage.HP treatment(300 MPa,2 min) also led to a reduction of aerobic bacterial count(APC) by 1.27 log cycles.Furthermore,the APC values of oysters treated by HP were always lower than those of the control samples during storage.Based on the organoleptic,biochemical and microbiological indicators,shelf life of 6-8 d for control and 12 d for HP-treated oysters could be expected.HP treatment showed great potential in oyster processing and preservation.

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

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

Researches on the Treatment of Phosphorous Wastewater with Oyster Shells

Based on the analysis of adsorptive features of oyster shells,the researches on the treatment of phosphorous wastewater with oyster shells and the effect of temperature on phosphorus removal were carried out.XRD was used to characterize the crystalline phases,and the main component of oyster shells was shown to be CaCO3.When the pretreatment temperature reached 800 ℃,some CaCO3 decomposed into CaO.As the temperature was further raised,CaO increased gradually.Via SEM testing,the oyster shell was a kind of natural porous materials.The pore wall partially collapsed from 550 to 900 ℃.No obvious porous structure was found at 900 ℃.However,without preheating,the oyster shell phosphorous removal material can not adsorb the phosphorus.Pretreatment made calcium activate,thus greatly increasing the absorption of phosphorus.

New Active Organic Substance in Oyster Shell Capable of Scavenging Oxygen Free Radicals with High Efficiency

A light purple organic active substance capable of scavenging hydroxyl radical（OH） with a high efficiency was extracted from Oyster shell at an extraction rate of 2.49%. It was found for the first time that this active substance may scavenge OH with the efficiency far higher than that of vitamin C. This active substance may scavenge also superoxide radical（O2） although the scavenging efficiency is far lower than that of vitamin C. Infrared spectrometry and routine chemical analysis primarily reveal that this active substance belongs to glycoprotein.

A new identification method for five species of oysters in genus Crassostrea from China based on high-resolution melting analysis

The high phenotypic plasticity in the shell of oysters presents a challenge during taxonomic and phylogenetie studies of these economically important bivalves. However, because DNA can exhibit marked differences among morphologically similar species, DNA barcoding offers a potential means for oyster identification. We analyzed the complete sequences of the cytochrome oxidase subunit I （COI） of five common Crassostrea species in China （including Hong Kong oyster C. hongkongensis, Jinjiang oyster C. ariakensis, Portuguese oyster C. angulata, Kumamoto oyster C. sikamea, and Pacific oyster C. gigas） and screened for distinct fragments. Using these distinct fragments on a high-resolution melting analysis platform, we developed an identification method that does not rely on species-specific PCR or fragment length polymorphism and is efficient, reliable, and easy to visualize. Using a single pair of primers （Oyster- COI-1）, we were able to successfully distinguish among the five oyster species. This new method provides a simple and powerful tool for the identification of oyster species.

Application of ICP-AES with Microwave Digestion to Detect Trace Elements in Oysters from Jiaozhou Bay,China

Using ICP-AES with microwave digestion, we determined the concentrations of 16 trace elements in oysters from six sampling points in Jiaozhou Bay.The distributions of the wholesome elements Zn, Fe and Mn, and heavy metals such as As,Cd,Hg and Pb were studied.The oysters collected are all rich in the wholesome trace elements,and the oysters from Licun River have the highest concentrations of the three wholesome trace elements.The concentrations of heavy metals in oysters from Licun River are the highest and those from Hongdao are the lowest. Compared with the domestic and foreign sea-areas,the heavy metal contents in the oysters from Jiaozhou Bay are less in amount than those from some developed countries, and more than those in Southeast China.This implies that the Jiaozhou Bay's oysters have been polluted by the environment to some extent.

Immune functional impacts of oyster peptide-based enteral nutrition formula (OPENF) on mice:a pilot study

Oyster peptides were produced from Crassostrea hongkongensis and used as a new protein source for the preparation of an oyster peptide-based enteral nutrition formula (OPENF). Reserpine-induced malabsorption mice and cyclophosphamide-induced immunosuppression mice were used in this study. OPENF powder is light yellow green and has a protein-fat-carbohydrate ratio of 16:9:75 with good solubility in water. A pilot study investigating immune functional impacts of the OPENF on mice show that the OPENF enhanced spleen lymphocyte proliferation and the activity of natural killer (NK) cells in BALB/c mice. Furthermore, OPENF can improve intestinal absorption, increase food utilization ratio, and maintain the normal physiological function of mice. These results suggest that oyster peptides could serve as a new protein source for use in enteral nutrition formula, but more importantly, also indicate that OPENF has an immunostimulating effect in mice.

Analysis of the Hydrodynamic Performance of an Oyster Wave Energy Converter Using Star-CCM+

A two-dimensional numerical Computational Fluid Dynamics(CFD)model is established on the basis of viscous CFD theory to investigate the motion response and power absorption performance of a bottom-hinged flap-type wave energy converter(WEC)under regular wave conditions.The convergence study of mesh size and time step is performed to ensure that wave height and motion response are sufficiently accurate.Wave height results reveal that the attenuation of wave height along the wave tank is less than 5%only if the suitable mesh size and time step are selected.The model proposed in this work is verified against published experimental and numerical models.The effects of mechanical damping,wave height,wave frequency,and water depth on the motion response,power generation,and energy conversion efficiency of the flap-type WEC are investigated.The selection of the appropriate mechanical damping of the WEC is crucial for the optimal extraction of wave power.The optimal mechanical damping can be readily predicted by using potential flow theory.It can then be verified by applying CFD numerical results.In addition,the motion response and the energy conversion efficiency of the WEC decrease as the incident wave height increases because the strengthened nonlinear effect of waves intensifies energy loss.Moreover,the energy conversion efficiency of theWEC decreases with increasing water depth and remains constant as the water depth reaches a critical value.Therefore,the selection of the optimal parameters during the design process is necessary to ensure that the WEC exhibits the maximum energy conversion efficiency.

Mass Selection for Growth Improvement in Black Shell Line of Pacific Oyster Crassostrea gigas

The Pacific oyster,Crassostrea gigas,naturally distributing along the coast of northwest Pacific,is one of the most important bivalve species due to its high economic value and fecundity.In China,we have initiated a selective breeding program on both shell color and growth rate of C.gigas since 2010.A black shell line was obtained through four-generation family selection.In this study,mass selection for growth improvement was conducted in the sixth generation and seventh generation of black shell lines.To assess the progress of potential genetic improvement,the progeny of two generations of black shell lines were selected to evaluate their shell heights via a 450-day farming experiment.As the results,after growing for 450 days,the sixth generation and seventh generation of selected lines were 9.03% and 11.42% larger than the control lines,respectively.During the grow-out stage,the genetic gain of two generations was 8.82%±0.18% and 11.54%±0.43%,respectively;and the corresponding realized heritability was 0.45±0.04 and 0.41±0.04,respectively.These results indicated that the mass selection for shell height achieved steady progress in the two generations of C.gigas.

An Efficient Method of Noroviruses Recovery from Oysters and Clams

Noroviruses (NoVs) are widespread causes of nonbacterial gastroenteritis. Outbreaks of NoVs caused diseases are commonly ascribed to the consumption of contaminated shellfish. The concentration and RNA extraction of NoVs are crucial steps of detecting NoVs in shellfish. This study aimed to select a simple, rapid and highly efficient recovery method of NoVs detection with real-time RT-PCR. Four methods of recovering GI.3 and GII.4 NoVs from spiked digestive tissues of oysters and clams, respectively, were compared, of them, the method involving proteinase K and PEG 8000 was found the most efficient. With this method, 9.3% and 13.1% of GI.3 and GII.4 NoVs were recovered from oysters and 9.6% and 12.3% of GI.3 and GII.4 NoVs were recovered from clams, respectively. This method was further used to detect NoVs in 84 oysters (Crassostrea gigas) and 86 clams (Ruditapes philippinarum) collected from 10 coastal cities in China from Jan. 2011 to Feb. 2012. The NoVs isolation rates were 10.47% of clams (9/86) and 7.14% of oysters (6/84). All the detected NoVs belonged to genotype GII. The NoVs recovery method selected is efficient for NoVs detection in oysters and clams.