AZM_3_(+)3.itax3D 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）是一类严峻的全球公共卫生威胁，亟需研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）的干预策略已展现出良好的应用前景。然而，多重耐药（Multidrug-resistant, MDR）病原体对上述疗法的代谢应答机制仍有待深入解析。本研究借助具备超高质量分辨率（＞240000）与亚2ppm质量精度的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健康粪便微生物群移植供体无菌粪便滤液（样本量n=4，供体平均年龄为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）的代谢改变进行了表征。