One of the problems with the processed meat products shelf life is the formation of yellow spots in sausages. The aim of this study was to isolate and identify the microbial agents causing yellow stain from sausage by molecular- and culture-based methods. For this purpose, 15 samples from yellow spot and separation areas of chicken sausages were sampled. A total of 44 colonies grown on the MRS, M17, YGC, and PDB culture medium were inoculated into pre-packaged cutting sausages to induce the yellow spot. Yellow spots appearance9 samples and they were characterized by both culture-based methods and sequencing of a 350 bp fragment from 16S rRNA V3 region and a 800bp fragment from 18S rRNA ITS4 region. The results of the culture-based and molecular assay showed that the main causes of the yellow spots in the chicken sausage packaged under vacuum were Enterococcus faecium, Enterococcusfaecalis, Streptococcus faecium and Candidasaitoana. In addition, one species of Enterococcus and one species of Candida were also identified at the genus level. Examination of yellowspotted samplesalso showed that the microorganisms that cause yellow spotting cause changes in pH, color indices L*, a* and b*, and syneresis compared to the control sample but the rate of change is only in the syneresis quantity and a* was significant.

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