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.

Near-infrared spectroscopy method for rapid proximate quantitative analysis of nutrient composition in Pacific oyster Crassostrea gigas

Glycogen,amino acids,fatty acids,and other nutrient components affect the flavor and nutritional quality of oysters.Methods based on near-infrared reflectance spectroscopy(NIRS)were developed to rapidly and proximately determine the nutrient content of the Pacific oyster Crassostreagigas.Samples of C.gigas from 19 costal sites were freeze-dried,ground,and scanned for spectral data collection using a Fourier transform NIR spectrometer(Thermo Fisher Scientific).NIRS models of glycogen and other nutrients were established using partial least squares,multiplication scattering correction first-order derivation,and Norris smoothing.The R_(C) values of the glycogen,fatty acids,amino acids,and taurine NIRS models were 0.9678,0.9312,0.9132,and 0.8928,respectively,and the residual prediction deviation(RPD)values of these components were 3.15,2.16,3.11,and 1.59,respectively,indicating a high correlation between the predicted and observed values,and that the models can be used in practice.The models were used to evaluate the nutrient compositions of 1278 oyster samples.Glycogen content was found to be positively correlated with fatty acids and negatively correlated with amino acids.The glycogen,amino acid,and taurine levels of C.gigas cultured in the subtidal and intertidal zones were also significantly different.This study suggests that C.gigas NIRS models can be a cost-effective alternative to traditional methods for the rapid and proximate analysis of various slaughter traits and may also contribute to future genetic and breeding-related studies in Pacific oysters.

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.

Assessing the effects of oyster/kelp weight ratio on water column properties: an experimental IMTA study at Sanggou Bay, China

Integrated Multi-Trophic Aquaculture(IMTA)is an effective method for sustainable aquaculture as species from different trophic levels could reduce negative effects from fed species in the environment.A proper proportion of different trophic species in an IMTA system could improve the aquaculture production and environmental sustainability.At present,research on the proper proportions for farming species is scarce.We investigated the effects of IMTA modes of oyster(Crassostrea gigas)and kelp(Saccharina japonica)in different weight ratios on water quality and carbonate system in a closed enclosure experiment for three days in the Sanggou Bay in Shandong Province,China,in December 2017.Nine collocation modes in oyster:kelp weight ratio were tested showing as 24:3,24:2,24:1,16:3,16:2,16:1,8:3,8:2,and 8:1.The water parameters were determined at 17:00 on Day 1(D1),and 6:00 and 17:00 on Days 2(D2)and 3(D3).As two-way ANOVA showed,all increased parameters(dissolved oxygen(DO),pH,chl a,the carbonate system and pCO2)were significantly related to oyster-kelp modes,and interaction between modes and time were also significant(P<0.05).On the 3th day,the 8:3 mode was the highest in DO,pH,chl a,CO32-(P<0.05),and dissolved inorganic carbon(DIC),HCO3-,CO2,and pCO2 were the lowest(P<0.05).According to previous references and the results of this study,the appropriate oyster:kelp proportion at the beginning of winter is from 8:2 to 8:3.The results of this study may help government to optimize the aquaculture structure of Sanggou Bay.

Response of phytoplankton to multispecies mariculture:a case study on the carrying capacity of shellfish in the Sanggou Bay in China

A muhispecies model for shellfish polycuhure in the Sanggou Bay in China used for large-scale long-line cultivation of the Chinese scallop Chlamysfarreri, the Pacific oyster Crassostrea gigas and the kelp Laminaria japonica is presented. The model includes key physical processes which are the transports of matter at the system boundary, and the main biological process that is the primary production and nutrients release from the bottom. By the model, the seasonal fluctuations of phytoplankton biomass and dissolved inorganic nitrogen（DIN） in 1994 are simulated. Furthermore, if the kelp culture scale is kept constant and the Chinese scallop and the Pacific oyster culture scales are adjusted, virtual shellfish farms are funded and responses of phytoplankton to the largescale shellfish culture are simulated. According to these simulated results, the room limitation, and the hypothesis that shellfish will not grow well if the phytoplankton biomass is less than 8.2 mg/m^3 , the expandable multiple of scallop culture k and that of oyster culture y are determined as k = -0.276 5y ＋4.690 5 and 0.133 3k ＋0.006 6y≤0.667 5, where, k （ or y） is equal to 1, the culture scale of scallop （ or oyster） is 8.8 x 109 individuals （or 66 ha, with a density of 59 ind./m^2 ）, and the kelp culture scale is 3 300 ha with a density of 12 ind./m^2.