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Background  As a kind of opportunist pathogen,  Staphylococcus xylosus (S. xylosus)  can cause mastitis. Antibiotics are widely used for treating infected animals and tylosin is a member of such group. Thus, the continuous use of antibiotics in dairy livestock enterprise will go a long way in increasing tylosin resistance. However, the mechanism of tylosin-resistant  S. xylosus  is not clear. Here, isobaric tag for relative and absolute quantitation (iTRAQ)-based quantitative proteomics methods was used to find resistance-related proteins.    Results  We compared the differential expression of  S. xylosus  in response to tylosin stress by iTRAQ. A total of 155 proteins (59 up-regulated, 96 down-regulated) with the fold-change of >1.2 or <0.8 ( p  value ≤0.05) were observed between the  S. xylosus  treated with 1/2 MIC (0.25 μg/mL) tylosin and the untreated  S. xylosus . Bioinformatic analysis revealed that these proteins play important roles in stress-response and transcription. Then, in order to verify the relationship between the above changed proteins and mechanism of tylosin-resistant  S. xylosus , we induced the tylosin-resistant  S. xylosus , and performed quantitative PCR analysis to verify the changes in the transcription proteins and the stress-response proteins in tylosin-resistant  S. xylosus  at the mRNA level .  The data displayed that ribosomal protein L23 ( rplw ), thioredoxin( trxA ) and Aldehyde dehydrogenase A( aldA-1 ) are up-regulated in the tylosin-resistant  S. xylosus , compared with the tylosin-sensitive strains.    Conclusion  Our findings demonstrate the important of stress-response and transcription in the tylosin resistance of  S. xylosus  and provide an insight into the prevention of this resistance, which would aid in finding new medicines .    Electronic supplementary material  The online version of this article (10.1186/s12917-019-1959-9) contains supplementary material, which is available to authorized users.

Comparative proteomic analysis reveals drug resistance of Staphylococcus xylosus ATCC700404 under tylosin stress