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With the purpose to investigate the relationship between spoilage and bacterial metabolism in lightly salted Mylopharyngodon piceus, Biolog GEN Ⅲ microplate was used to understand the carbon metabolism of predominant spoilage bacteria, Staphylococcus xylosus, at 5℃, 15℃, 25℃ and 33℃, respectively, and the growth curve of Staphylococcus xylosus was fitted with the modified Gompetz model. In addition, the metabolic rate was measured by using average well color development (AWCD) as indicator to describe its dynamic properties of carbon utilization. The results showed that saccharides, carboxylic acids and amino acids were utilized by Staphylococcus xylosus with degressive utilization ratio at the same temperature. Among carbon substrates, monosaccharides (e.g., glucose and mannose), disaccharides (e.g., sucrose and trehalose), polysaccharides and sugar derivatives (e.g., N-Acetyl-β-D-Mannose amine and β-Methyl-D-Glucoside), amino acids (e.g., glutamate and serine) and carboxylic acids (e.g., L-lactate) were dramatically metabolized. The lag phase of Staphylococcus xylosus was the shortest at 25℃, thus the exponential phase was reached fast. Overall, the present study focused on carbon utilization and metabolism of Staphylococcus xylosus at different temperatures to provide a theoretical basis for inhibition of microbial activity by optimizing formula and extension of shelf life of lightly salted Mylopharyngodon piceus. It was proposed that Biolog GEN Ⅲ microplate could be applied to establishment of bacterial predictive model combining simultaneous investigation on microbial activity of carbon sources utilization and metabolism.