The spoilage microbiota in water-boiled salted duck during storage at 4 degrees C was determined using culture-dependent and independent methods. Analysis of the denaturing gradient gel electrophoresis (DGGE) patterns of PCR amplicons targeting the V3 region of the 16S rDNA and sequencing of the bands allowed profiling of the microbiota present in the duck. Community DNA extracts were prepared directly from water-boiled salted duck and from culturable bacterial fractions harvested from both MRS and PCA media. The spoilage bacteria mainly consisted of Staphylococcus saprophyticus, Macrococcus caseolyticus, Weissella, Halomonas sp. or Cobetia sp., and Exiguobacterium sp. based on sequencing and homology search of the DGGE bands. It appeared that both the bacterial counts and diversity increased during storage time. By plating method, bacterial counts in MRS agar increased from 10(4) to 10(8) CFU/g from day 1 to 10, while total bacterial counts in PCA agar reached 10(9) CFU/g after 10 d. Total of 14 strains isolated from PCA and MRS agar were identified as M. caseolyticus (2), S. saprophyticus (7), S. sciuri (1), W. paramesenteroides (2), and W. confusa (2) by 16S rDNA sequencing. The identification of the spoilage-related microbiota is helpful to better understand the bacteria ecology in water-boiled salted duck and may lead to the discovery of appropriate preservation strategies.