AP_2_(+)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).

抗菌药物耐药性（Antimicrobial Resistance, AMR）是当前全球健康领域面临的紧迫威胁，亟需寻找传统抗生素之外的创新替代方案。其中，噬菌体疗法与基于粪便微生物群移植（faecal microbiota transplantation, FMT）的干预措施已成为颇具前景的策略。然而，多药耐药（multidrug-resistant, MDR）病原体对这些疗法的代谢应答机制仍知之甚少。本研究应用先进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——该技术具备超高质量分辨率（>240000）与低于2 ppm的质量准确度——对致病性大肠杆菌菌株（肠聚集性大肠杆菌Enteroaggregative, EAEC；肠侵袭性大肠杆菌Enteroinvasive, EIEC）在经裂解性噬菌体vB_EcoM_SVMS_56、噬菌体-阿奇霉素联合疗法及健康FMT供体（n=4；平均年龄33.75±2.38岁）的无菌粪便滤液处理后的代谢变化进行了分析。