| Campylobacter jejuni is one of the most common food-borne pathogens causing bacterial diarrhea and Guillain-Barres syndrome in human. Amphenicols including chloramphenicol (CAP), florfenicol (FFC) and thiamphenine played an important role in the prevention of C. jejuni and other Gram negative bacteria infection in veterinary medicine, except for CAP. However, owing to its wide application, the resistance of C. jejuni was very common and the transmission of drug-resistant C. jejuni will bring enormous pressure in the treatment of human clinical Campylobactiosis. Transcriptomics, proteomics and metabolomics as the new "omics" technologies can deciper specific biological processes and regulation mechanism occurred in an organism from the overall level. In this study, we aimed to carry out multi-omics analysis and explore the possible molecular mechanism of antibiotic resistance across C. jejuni susceptible strains and CAP-, FFC-resistant strains.Strain-specific transcriptome of C. jejuni susceptible and mutant strains was analyzed by using RNA-Seq. A total of1347unigenes were obtained in C. jejuni control strains using its reference genome to analyze the yield and quality of assembly. A total of290differential expression genes (DEGs)(182genes up-regulated,108genes down-regulated) were identified in CAP-resistant mutants and these DEGs were involved in many metabolic pathways such as metabolic pathway, biosynthesis of secondary metabolites, microbial metabolism in diverse environments, ribosome and flagellar assembly.93DEGs (32genes up-regulated,61genes down-regulated) were found to mediate FFC resistance in C. jejuni. KEGG pathway analysis showed that metabolic pathway, biosynthesis of secondary metabolites, microbial metabolism in diverse environments, ribosome and nitrogen metabolism were related to FFC-resistant C. jejuni.Then, a filter-aided sample preparation was combined with SWATH-MS data independent acquisition mode for the label-free quantitative proteomic analysis of C. jejuni susceptible strains and CAP-, FFC-resistant mutants. A total of197differential proteins were identified in CAP-resistant mutants and GO functional annotation showed that these proteins were located in cell part, membrane and organelles and mainly involved in metabolism, cellular process and single-organism process. Pathway analysis showed that metabolic pathway, biosynthesis of secondary metabolites, biosynthesis of amino acid and ABC transporters were related to CAP-resistance in C. jejuni. There were84differential proteins related to FFC-resistant C. jejuni. These protein were located in cell part, organelles, cell membrane and involved in cell process, single-organism process, metabolism and colonization process. Pathway analysis showed that metabolic pathway, biosynthesis of secondary metabolites, biosynthesis of amino acid, two-component system and flagellar assembly were related to FFC-resistance in C. jejuni. There were34differential protein (18protein up-regulated,16protein down-regulated) which simultaneously appeared in CAP-and FFC-resistant mutants.Untargeted metabolomic profiling of amphenicol-resistant C. jejuni by ultra-high performance liquid chromatography-time of flight mass spectrometry was performed to analyze the small molecule biomarkers of drug resistance. Principal component analysis (PCA) and partial least squares discriminant analysis were applied to classify C. jejuni samples. Up to41differential metabolites involved in glycerophospholipid metabolism, sphingolipid metabolism and fatty acid metabolism were observed in CAP-resistant mutant strain of C. jejuni. A panel of40features were identified in florfenicol-resistant mutants, demonstrating changes in glycerophospholipid metabolism, sphingolipid metabolism, and tryptophan metabolism. There were27differential metabolites which simultaneously appeared in CAP-and FFC-resistant mutants.In summary, the occurring of antibiotic resistance in amphenicols-resistant C. jejuni mainly referred to ribosome, flagellar assembly, ABC transporters and chemotaxis protein by comprehensive analysis of transcriptomics, proteomics and metabolomics. In addition, glycerophospholipid metabolism was disturbed in mediating the process of amphenicols resistance in C. jejuni. This study demonstrated that the multi-omics technologies platform was a promising and valuable tool to generate new insights into the drug-resistant mechanism of C. jejuni. Furthermore, it is helpful to provide some scientific reference for the prevention of the antibiotic resistance and the development of the antibiotic agents. |
PDF Full Text Request