College of Veterinary Medicine, Southwest University, Chongqing, China.
National Center of Technology Innovation for Pigs, Chongqing, China.
Front Cell Infect Microbiol. 2024 Aug 15;14:1406429. doi: 10.3389/fcimb.2024.1406429. eCollection 2024.
Dairy mastitis is one of the most common diseases in dairy farming, and the formation of pathogenic bacteria biofilms may be an important reason why traditional antibiotic therapy fails to resolve some cases of dairy mastitis. We isolated and identified three strains of were with strong biofilm forming ability from dairy cow mastitis samples from Chongqing dairy farms in China. In order to investigate the effect of novel anti-biofilm peptide CRAMP-34 on biofilms, the anti-biofilm effect was evaluated by crystal violet staining, biofilms viable bacteria counting and confocal laser scanning microscopy (CLSM). In addition, transcriptome sequencing analysis, qRT-PCR and phenotypic verification were used to explore the mechanism of its action. The results showed that CRAMP-34 had a dose-dependent eradicating effect on biofilms. Transcriptome sequencing analysis showed that 36 differentially expressed genes (11 up-regulated and 25 down-regulated) were detected after the intervention with the sub-inhibitory concentration of CRAMP-34. These differentially expressed genes may be related to enzyme synthesis, fimbriae, iron uptake system, capsular polysaccharide and other virulence factors through the functional analysis of differential genes. The results of subsequent bacterial motility and adhesion tests showed that the motility of were enhanced after the intervention of CRAMP-34, but there was no significant change in adhesion. It was speculated that CRAMP-34 may promote the dispersion of biofilm bacteria by enhancing the motility of biofilm bacteria, thereby achieving the effect of eradicating biofilms. Therefore, these results, along with our other previous findings, suggest that CRAMP-34 holds promise as a new biofilm eradicator and deserves further research and development.
奶牛乳腺炎是奶牛养殖业中最常见的疾病之一,而病原菌生物膜的形成可能是传统抗生素治疗无法解决某些奶牛乳腺炎病例的一个重要原因。我们从中国重庆奶牛场的奶牛乳腺炎样本中分离并鉴定出三株具有较强生物膜形成能力的 。为了研究新型抗生物膜肽 CRAMP-34 对 生物膜的影响,通过结晶紫染色、生物膜活菌计数和共聚焦激光扫描显微镜(CLSM)评估了抗生物膜作用。此外,还通过转录组测序分析、qRT-PCR 和表型验证来探索其作用机制。结果表明,CRAMP-34 对 生物膜具有剂量依赖性的清除作用。转录组测序分析显示,在亚抑菌浓度的 CRAMP-34 干预后,检测到 36 个差异表达基因(11 个上调和 25 个下调)。通过对差异基因的功能分析,这些差异表达基因可能与酶合成、菌毛、铁摄取系统、荚膜多糖和其他毒力因子有关。随后的细菌运动性和黏附性测试结果表明,CRAMP-34 干预后 的运动性增强,但黏附性没有明显变化。推测 CRAMP-34 可能通过增强生物膜细菌的运动性来促进生物膜细菌的分散,从而达到清除生物膜的效果。因此,这些结果以及我们之前的其他发现表明,CRAMP-34 有希望成为一种新型的生物膜清除剂,值得进一步研究和开发。