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• The effects of depuration salinity and duration on preservation were evaluated. • The mortality increased with 72 h depuration and 20% salinity fluctuation. • The SWATH-MS-based proteomic was developed to analysis protein changes. • 381 different expression proteins (DEPs) in total uncovered by proteomics analysis. • Most DEPs were relevant to amino acid metabolism. Depuration and anhydrous living-preservation are two important and consecutive steps during the circulation of Pacific oyster (Crassostrea gigas), and two main factors in depuration, salinity and duration, are pivotal to posterior anhydrous living-preservation of C. gigas. In this work, the comprehensive effects of depuration salinity (26–38 g/L) and duration (0–72 h) on anhydrous living-preservation at 4 °C for 7 days were investigated in regard of mortality, biochemical indexes (fatty acids profile analysis, glycogen) and proteome changes as well. The results showed that the mortality of C. gigas increased obviously with 72 h depuration and especially with 20 % salinity fluctuation, concomitantly accompanying metabolism disorder. Furthermore, alterations in salinity and duration resulted in 381 different expression proteins (DEPs), which were gotten more involved in the pathways related to amino acid metabolism. Taken together, it was suggested that duration < 48 h and salinity fluctuations lower than 10 % were the preferred conditions for anhydrous living-preservation. [ABSTRACT FROM AUTHOR]

• Proteomic analysis revealed differentiating proteins (DPs) in C. gigas. • Levels of DPs associated with glycolysis increased with salinity stress. • Levels of DPs associated with ECM-receptor interaction pathway increased. • Depuration in 38 g/L salinity was detrimental to oyster quality. • Depuration salinity fluctuation should be controlled within ± 10% of production area. Salinity stress during depuration of Pacific oysters (Crassostrea gigas) leads to degradation in quality; therefore, an understanding of the molecular mechanisms regulating dynamic changes during depuration is needed. Here, C. gigas was depurated for 72 h at salinities ranging from 26 to 38 g/L, a ± 10–20% fluctuation from that in the production area, and the gill proteomes were analyzed by sequential window acquisition of all theoretical fragment ion mass spectra (SWATH-MS). Of the 1218 proteins analyzed, 241 were differentiating proteins (DPs). Salinity stress led to increased levels of DPs associated with glycolysis and the extracellular matrix–receptor interaction pathway, and decreased levels of DPs associated with the citric acid cycle, lipid metabolism, genetic information processing, and cell transformation, especially in oysters exposed to 38 g/L salinity (+20%). Controlling salinity fluctuation within ± 10% of the production area during depuration was conducive to maintaining quality in C. gigas. [ABSTRACT FROM AUTHOR]