D_4_(-)1.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，亟需研发传统抗生素的创新替代疗法。其中，噬菌体疗法与基于粪便微生物群移植（Fecal microbiota transplantation, FMT）的干预手段已展现出可观的应用前景。然而，多重耐药（Multidrug-resistant, MDR）病原体对上述治疗策略的代谢应答机制仍有待阐明。本研究借助具备超高质量分辨率（＞240000）与亚2ppm质量精度的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4名健康粪便移植供体（平均年龄33.75±2.38岁）的无菌粪便滤液处理后的致病性大肠杆菌菌株——聚集性大肠杆菌（Enteroaggregative, EAEC）与侵袭性大肠杆菌（Enteroinvasive, EIEC）的代谢改变进行了表征。