A Novel Bacteriophage Targeting <i>mcr-9 Enterobacter kobei</i> with Potential Application in Fresh Leafy Greens

The presence of opportunistic pathogens, such as the <i>Enterobacter cloacae</i> 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 these ar...

Full description

Saved in:
Bibliographic Details
Main Authors: Alberto Pintor-Cora, Andrea Carpintero, Ángel Alegría, Apostolos Giannis, Teresa-María Lopez-Díaz, Jesús A. Santos, Jose M. Rodríguez-Calleja
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Applied Microbiology
Subjects:
Online Access:https://www.mdpi.com/2673-8007/5/1/25
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849340876662243328
author Alberto Pintor-Cora
Andrea Carpintero
Ángel Alegría
Apostolos Giannis
Teresa-María Lopez-Díaz
Jesús A. Santos
Jose M. Rodríguez-Calleja
author_facet Alberto Pintor-Cora
Andrea Carpintero
Ángel Alegría
Apostolos Giannis
Teresa-María Lopez-Díaz
Jesús A. Santos
Jose M. Rodríguez-Calleja
author_sort Alberto Pintor-Cora
collection DOAJ
description The presence of opportunistic pathogens, such as the <i>Enterobacter cloacae</i> 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 these are 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 cattle farm sewage. Belonging to the <i>Seunavirus</i> genus, FENT2 did not carry genes associated with lysogenic cycle, antimicrobial resistance, or virulence factors. The phage demonstrated lytic activity against the host strain <i>E. kobei</i> AG07E, which harbored the <i>mcr-9</i> gene, exhibiting a narrow host range that also included <i>E. ludwigii</i> strains. In vitro assays using BioTrac (SY-LAB) impedance technology confirmed the sustained lytic activity of FENT2 under food-related stress conditions, including pH levels from 5 to 7 and NaCl concentrations up to 2%. Furthermore, FENT2 demonstrated bactericidal potential on lettuce leaves, achieving 1 log reduction in bacterial counts of the host strain after 30 min immersion treatment. These findings highlight FENT2 as a promising candidate for biocontrol applications, offering a sustainable alternative to conventional decontamination methods for reducing antimicrobial-resistant ECC contamination in fresh produce.
format Article
id doaj-art-6dbcc2ee75c5403a9e56cefb99017ce0
institution Kabale University
issn 2673-8007
language English
publishDate 2025-02-01
publisher MDPI AG
record_format Article
series Applied Microbiology
spelling doaj-art-6dbcc2ee75c5403a9e56cefb99017ce02025-08-20T03:43:47ZengMDPI AGApplied Microbiology2673-80072025-02-01512510.3390/applmicrobiol5010025A Novel Bacteriophage Targeting <i>mcr-9 Enterobacter kobei</i> with Potential Application in Fresh Leafy GreensAlberto Pintor-Cora0Andrea Carpintero1Ángel Alegría2Apostolos Giannis3Teresa-María Lopez-Díaz4Jesús A. Santos5Jose M. Rodríguez-Calleja6Department of Food Hygiene and Food Technology, Veterinary Faculty, Universidad de León, 24071 León, SpainDepartment of Food Hygiene and Food Technology, Veterinary Faculty, Universidad de León, 24071 León, SpainDepartment of Food Hygiene and Food Technology, Veterinary Faculty, Universidad de León, 24071 León, SpainSchool of Chemical and Environmental Engineering, Technical University of Crete, University Campus, 73100 Chania, GreeceDepartment of Food Hygiene and Food Technology, Veterinary Faculty, Universidad de León, 24071 León, SpainDepartment of Food Hygiene and Food Technology, Veterinary Faculty, Universidad de León, 24071 León, SpainDepartment of Food Hygiene and Food Technology, Veterinary Faculty, Universidad de León, 24071 León, SpainThe presence of opportunistic pathogens, such as the <i>Enterobacter cloacae</i> 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 these are 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 cattle farm sewage. Belonging to the <i>Seunavirus</i> genus, FENT2 did not carry genes associated with lysogenic cycle, antimicrobial resistance, or virulence factors. The phage demonstrated lytic activity against the host strain <i>E. kobei</i> AG07E, which harbored the <i>mcr-9</i> gene, exhibiting a narrow host range that also included <i>E. ludwigii</i> strains. In vitro assays using BioTrac (SY-LAB) impedance technology confirmed the sustained lytic activity of FENT2 under food-related stress conditions, including pH levels from 5 to 7 and NaCl concentrations up to 2%. Furthermore, FENT2 demonstrated bactericidal potential on lettuce leaves, achieving 1 log reduction in bacterial counts of the host strain after 30 min immersion treatment. These findings highlight FENT2 as a promising candidate for biocontrol applications, offering a sustainable alternative to conventional decontamination methods for reducing antimicrobial-resistant ECC contamination in fresh produce.https://www.mdpi.com/2673-8007/5/1/25bacteriophagesopportunistic pathogens<i>Enterobacter cloacae</i> complexfresh producevegetables
spellingShingle Alberto Pintor-Cora
Andrea Carpintero
Ángel Alegría
Apostolos Giannis
Teresa-María Lopez-Díaz
Jesús A. Santos
Jose M. Rodríguez-Calleja
A Novel Bacteriophage Targeting <i>mcr-9 Enterobacter kobei</i> with Potential Application in Fresh Leafy Greens
Applied Microbiology
bacteriophages
opportunistic pathogens
<i>Enterobacter cloacae</i> complex
fresh produce
vegetables
title A Novel Bacteriophage Targeting <i>mcr-9 Enterobacter kobei</i> with Potential Application in Fresh Leafy Greens
title_full A Novel Bacteriophage Targeting <i>mcr-9 Enterobacter kobei</i> with Potential Application in Fresh Leafy Greens
title_fullStr A Novel Bacteriophage Targeting <i>mcr-9 Enterobacter kobei</i> with Potential Application in Fresh Leafy Greens
title_full_unstemmed A Novel Bacteriophage Targeting <i>mcr-9 Enterobacter kobei</i> with Potential Application in Fresh Leafy Greens
title_short A Novel Bacteriophage Targeting <i>mcr-9 Enterobacter kobei</i> with Potential Application in Fresh Leafy Greens
title_sort novel bacteriophage targeting i mcr 9 enterobacter kobei i with potential application in fresh leafy greens
topic bacteriophages
opportunistic pathogens
<i>Enterobacter cloacae</i> complex
fresh produce
vegetables
url https://www.mdpi.com/2673-8007/5/1/25
work_keys_str_mv AT albertopintorcora anovelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT andreacarpintero anovelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT angelalegria anovelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT apostolosgiannis anovelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT teresamarialopezdiaz anovelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT jesusasantos anovelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT josemrodriguezcalleja anovelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT albertopintorcora novelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT andreacarpintero novelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT angelalegria novelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT apostolosgiannis novelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT teresamarialopezdiaz novelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT jesusasantos novelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens
AT josemrodriguezcalleja novelbacteriophagetargetingimcr9enterobacterkobeiiwithpotentialapplicationinfreshleafygreens