Abstract
PURPOSE: Effective decolonization strategies for intestinal carriers of carbapenem-resistant Enterobacterales are essential to prevent severe life-threatening infections. In this work, we established gut colonization in Zophobas morio larvae (ZmL) using an OXA-48-producing Salmonella enterica ST198 strain (Sk-1) and assessed the commercial INTESTI bacteriophage cocktail (INTESTIbc) for decolonization. METHODS: ZmL were fed with food contaminated with Sk-1 (INTESTIbc-susceptible) for 3 days and then maintained on a non-contaminated diet until day 14 (T14). At T3, ZmL were grouped in untreated, dPBS- or INTESTIbc-treated (oral force-feeding on T3 and T5). At specified intervals, ZmL were sampled for quantification and characterization of Sk-1 (antibiotic/INTESTIbc susceptibility and whole-genome sequencing). ZmL microbiota was also investigated by 16S rRNA amplicon sequencing. RESULTS: ZmL were rapidly colonized by Sk-1 across all groups (T3: 4.3 × 10(6) CFU/mL). Untreated and dPBS-treated larvae remained consistently colonized (T10: 3.4-9.1 × 10(4) CFU/mL; T14: 2.9-5.9 × 10(4) CFU/mL), whereas INTESTIbc treatment induced a significant Sk-1 regrowth (T10: 4.0 × 10(6) CFU/mL; P < 0.05 vs. controls). Sk-1 strains recovered under different conditions between T7 and T14 did not show phenotypic and genotypic changes. Bacteriophages administration resulted in reduced relative abundance of potential bacterial competitors of Sk-1 (i.e., Pseudocitrobacter). CONCLUSIONS: ZmL can be used as a new in vivo model of intestinal colonization with S. enterica. However, INTESTIbc administration failed to achieve decolonization and instead promoted hazardous overgrowth of the inoculated pathogen. These findings highlight the need for further investigations to clarify the therapeutic potential or possible risks of broad-spectrum bacteriophage cocktails against intestinal infections/colonization caused by hyperepidemic S. enterica clones.