4.6 Article

Transcriptomic Changes and satP Gene Function Analysis in Pasteurella multocida with Different Levels of Resistance to Enrofloxacin

Journal

VETERINARY SCIENCES
Volume 10, Issue 4, Pages -

Publisher

MDPI
DOI: 10.3390/vetsci10040257

Keywords

bovine respiratory disease; Pasteurella multocida; drug resistance; antibiotic tolerance; satP

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The study found that Pasteurella multocida, a major pathogen of bovine respiratory disease, showed high resistance to the commonly used antibiotic enrofloxacin. Transcriptome sequencing revealed that the expression of the satP gene changed significantly with increasing drug resistance. Further experiments confirmed that the satP gene is related to the resistance and pathogenicity of Pasteurella multocida and may serve as a target for synergistic effects with enrofloxacin.
Simple Summary Pasteurella multocida is a major pathogen of bovine respiratory disease, which is resistant to many of the commonly used antibiotics. We found that enrofloxacin has shown a high drug resistance in clinical treatment of Pasteurella multocida infection. In order to better understand the resistance mechanism of Pasteurella multocida to enrofloxacin, we isolated PmS and PmR strains with the same PFGE typing in vitro, and artificially induced PmR to obtain the highly resistant phenotype, PmHR. Then, transcriptome sequencing of clinically isolated sensitive strains, resistant and highly drug-resistant strains, treated with enrofloxacin at sub-inhibitory concentrations, were performed. The satP gene, of which the expression changed significantly with the increase in drug resistance, was screened. In order to further confirm the function of this gene, we constructed the deleted and complemented strains of satP, and further analyzed the function of the satP gene. After satP gene deletion, the resistance of Pasteurella multocida was obviously lower than that of wild-type strains in vitro, and the pathogenicity of Pasteurella multocida was reduced by about 400 times. We found that the satP gene is related to the tolerance and pathogenicity of Pasteurella multocida, and can be used as a target of enrofloxacin synergistic effect. Pasteurella multocida (Pm) is one of the major pathogens of bovine respiratory disease (BRD), which can develop drug resistance to many of the commonly used antibiotics. Our earlier research group found that with clinical use of enrofloxacin, Pm was more likely to develop drug resistance to enrofloxacin. In order to better understand the resistance mechanism of Pm to enrofloxacin, we isolated PmS and PmR strains with the same PFGE typing in vitro, and artificially induced PmR to obtain the highly resistant phenotype, PmHR. Then transcriptome sequencing of clinically isolated sensitive strains, resistant and highly drug-resistant strains, treated with enrofloxacin at sub-inhibitory concentrations, were performed. The satP gene, of which the expression changed significantly with the increase in drug resistance, was screened. In order to further confirm the function of this gene, we constructed a satP deletion (Delta Pm) strain using suicide vector plasmid pRE112, and constructed the C-Pm strain using pBBR1-MCS, and further analyzed the function of the satP gene. Through a continuously induced resistance test, it was found that the resistance rate of Delta Pm was obviously lower than that of Pm in vitro. MDK99, agar diffusion and mutation frequency experiments showed significantly lower tolerance of Delta Pm than the wild-type strains. The pathogenicity of Delta Pm and Pm was measured by an acute pathogenicity test in mice, and it was found that the pathogenicity of Delta Pm was reduced by about 400 times. Therefore, this study found that the satP gene was related to the tolerance and pathogenicity of Pm, and may be used as a target of enrofloxacin synergistic effect.

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