D_7_(+)3.itm3D OrbiSIMS Metabolic Profiling of E. coli Reveals Effects of FMT- Derived Faecal Filtrate and Phage-Azithromycin on Amino Acids and Lipid Pathways.

Antimicrobial resistance (AMR) represents a pressing global health threat, necessitating innovative alternatives to traditional antibiotics. Among these, bacteriophage therapy and faecal microbiota transplantation (FMT)-based interventions have emerged as promising strategies. However, the metabolic responses of multidrug-resistant (MDR) pathogens to these therapies remain poorly understood. In this study, we applied advanced 3D Orbitrap Secondary Ion Mass Spectrometry (3D OrbiSIMS) metabolomics, offering ultra-high mass resolution (>240,000) and sub-2 ppm mass accuracy, to profile metabolic alterations in pathogenic Escherichia coli strains, Enteroaggregative (EAEC) and Enteroinvasive (EIEC), following treatment with lytic phage vB_EcoM_SVMS_56, phage–azithromycin combination therapy, and sterile faecal filtrates from healthy FMT donors (n = 4; mean age 33.75 ± 2.38 years).

抗菌素耐药性 (Antimicrobial resistance, AMR) 是亟待应对的全球性公共卫生威胁，亟需开发传统抗生素以外的创新替代疗法。其中，噬菌体疗法与基于粪便微生物群移植 (faecal microbiota transplantation, FMT) 的干预手段已成为颇具前景的治疗策略。然而，多重耐药病原体 (multidrug-resistant, MDR) 对上述疗法的代谢应答机制仍有待深入解析。本研究采用具备超高质量分辨率 (>240000) 与亚2ppm质量精度的先进三维轨道阱二次离子质谱 (3D Orbitrap Secondary Ion Mass Spectrometry, 3D OrbiSIMS) 代谢组学技术，对经裂解性噬菌体vB_EcoM_SVMS_56、噬菌体-阿奇霉素联合疗法以及健康FMT供体的无菌粪便滤液 (n=4；平均年龄33.75±2.38岁) 处理后的致病性大肠杆菌菌株——肠聚集性大肠杆菌 (Enteroaggregative, EAEC) 与肠侵袭性大肠杆菌 (Enteroinvasive, EIEC) 的代谢变化进行了表征。