D_1_(-)1.itmx3D 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).抗菌药物耐药性（Antimicrobial resistance, AMR）是一类亟待应对的全球公共卫生威胁，亟需开发传统抗生素之外的创新替代疗法。其中，噬菌体疗法与基于粪便微生物群移植（Fecal microbiota transplantation, FMT）的干预手段已成为极具潜力的治疗策略。然而，目前对于多重耐药（Multidrug-resistant, MDR）病原体针对此类疗法的代谢响应仍知之甚少。本研究采用具备超高质量分辨率（>240,000）且质量准确度优于2 ppm的先进3D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来自4名健康粪便微生物群移植供体（供体平均年龄为33.75±2.38岁）的无菌粪便滤液处理后的肠聚集性大肠杆菌（Enteroaggregative, EAEC）与肠侵袭性大肠杆菌（Enteroinvasive, EIEC）这两类致病性大肠杆菌菌株的代谢变化进行了表征。