Phage-Based Biocontrol of Antibiotic-Resistant <i>Enterobacter cloacae </i>Complex in Fresh Produce

The presence of opportunistic pathogens, such as the Enterobacter cloacae complex (ECC), in fresh vegetables poses a significant health risk, particularly amid the ongoing antibiotic resistance crisis. Traditional chemical decontamination methods are often ineffective and associated with issues such as cross-resistance between antibiotics and biocides, highlighting the need for alternative approaches. This study describes the isolation of a novel phage, FENT2, with anti-ECC activity, obtained from wastewater. Belonging to the Seunavirus genus, FENT2 did not carry genes associated with lysogenic cycle, antimicrobial resistance, or virulence factors. The phage demonstrated lytic activity against the host strain E. kobei AG07E strain, which harbors the mcr-9 gene. FENT2 exhibited a broad host range, infecting 82.6 % of ECC strains tested. In vitro assays using BioTrac (SY-LAB) technology confirmed the phage&#039;s efficacy, maintaining lytic activity under food-related stress conditions (pH levels from 5 to 7 and % NaCl up to 2). Furthermore, FENT2 demonstrated bactericidal potential on lettuce leaves, achieving an approximate 1 log reduction in bacterial counts of the host strain after immersion treatment. This study highlights FENT2 as a promising biocontrol agent in the food industry, providing an effective and sustainable alternative to traditional chemical decontamination methods.