The Virulence Factor LLO of <i>Listeria monocytogenes</i> Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect

<i>Listeria monocytogenes</i> is a life-threatening bacterial foodborne pathogen that can persist in food-processing facilities for years. Although phages can control <i>L. monocytogenes</i> during food production, phage-resistant bacterial subpopulations can regrow in phage-...

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Main Authors: Banhong Liu, Mei Bai, Wuxiang Tu, Yanbin Shen, Jingxin Liu, Zhenquan Yang, Hongduo Bao, Qingli Dong, Yangtai Liu, Ran Wang, Hui Zhang, Liangbing Hu
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Foods
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Online Access:https://www.mdpi.com/2304-8158/14/15/2554
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author Banhong Liu
Mei Bai
Wuxiang Tu
Yanbin Shen
Jingxin Liu
Zhenquan Yang
Hongduo Bao
Qingli Dong
Yangtai Liu
Ran Wang
Hui Zhang
Liangbing Hu
author_facet Banhong Liu
Mei Bai
Wuxiang Tu
Yanbin Shen
Jingxin Liu
Zhenquan Yang
Hongduo Bao
Qingli Dong
Yangtai Liu
Ran Wang
Hui Zhang
Liangbing Hu
author_sort Banhong Liu
collection DOAJ
description <i>Listeria monocytogenes</i> is a life-threatening bacterial foodborne pathogen that can persist in food-processing facilities for years. Although phages can control <i>L. monocytogenes</i> during food production, phage-resistant bacterial subpopulations can regrow in phage-treated environments. In this study, an <i>L. monocytogenes hly</i> defective strain, NJ05-Δ<i>hly</i>, was produced, which considerably regulated the interactions between <i>L. monocytogenes</i> and phages. Specifically, we observed a 76.92-fold decrease in the efficiency of plating of the defective strain following infection with the <i>Listeria</i> phage vB-LmoM-NJ05. The lytic effect was notably diminished at multiplicities of infection of 1 and 10. Furthermore, the inactivation of LLO impaired biofilm formation, which was completely suppressed and eliminated following treatment with 10<sup>8</sup> PFU/mL of phage. Additionally, phages protected cells from mitochondrial membrane damage and the accumulation of mitochondrial reactive oxygen species induced by <i>L. monocytogenes</i> invasion. Transcriptomic analysis confirmed these findings, revealing the significant downregulation of genes associated with phage sensitivity, pathogenicity, biofilm formation, and motility in <i>L. monocytogenes</i>. These results underscore the vital role of LLO in regulating the pathogenicity, phage susceptibility, and biofilm formation of <i>L. monocytogenes</i>. These observations highlight the important role of virulence factors in phage applications and provide insights into the potential use of phages for developing biosanitizers.
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publishDate 2025-07-01
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spelling doaj-art-0c21560d3ef84d1ca5b151e6f6b2f6832025-08-20T03:36:03ZengMDPI AGFoods2304-81582025-07-011415255410.3390/foods14152554The Virulence Factor LLO of <i>Listeria monocytogenes</i> Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic EffectBanhong Liu0Mei Bai1Wuxiang Tu2Yanbin Shen3Jingxin Liu4Zhenquan Yang5Hongduo Bao6Qingli Dong7Yangtai Liu8Ran Wang9Hui Zhang10Liangbing Hu11Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of MOST, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of MOST, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of MOST, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaSchool of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, ChinaSchool of Food Science and Engineering, Yangzhou University, Yangzhou 225001, ChinaSchool of Food Science and Engineering, Yangzhou University, Yangzhou 225001, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of MOST, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaSchool of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, ChinaSchool of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of MOST, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaJiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of MOST, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, ChinaSchool of Food and Biological Engineering, Shaanxi University of Science and Technology, Xi’an 710021, China<i>Listeria monocytogenes</i> is a life-threatening bacterial foodborne pathogen that can persist in food-processing facilities for years. Although phages can control <i>L. monocytogenes</i> during food production, phage-resistant bacterial subpopulations can regrow in phage-treated environments. In this study, an <i>L. monocytogenes hly</i> defective strain, NJ05-Δ<i>hly</i>, was produced, which considerably regulated the interactions between <i>L. monocytogenes</i> and phages. Specifically, we observed a 76.92-fold decrease in the efficiency of plating of the defective strain following infection with the <i>Listeria</i> phage vB-LmoM-NJ05. The lytic effect was notably diminished at multiplicities of infection of 1 and 10. Furthermore, the inactivation of LLO impaired biofilm formation, which was completely suppressed and eliminated following treatment with 10<sup>8</sup> PFU/mL of phage. Additionally, phages protected cells from mitochondrial membrane damage and the accumulation of mitochondrial reactive oxygen species induced by <i>L. monocytogenes</i> invasion. Transcriptomic analysis confirmed these findings, revealing the significant downregulation of genes associated with phage sensitivity, pathogenicity, biofilm formation, and motility in <i>L. monocytogenes</i>. These results underscore the vital role of LLO in regulating the pathogenicity, phage susceptibility, and biofilm formation of <i>L. monocytogenes</i>. These observations highlight the important role of virulence factors in phage applications and provide insights into the potential use of phages for developing biosanitizers.https://www.mdpi.com/2304-8158/14/15/2554<i>Listeria hemolysin</i>biofilmphage susceptibilitytranscriptomics
spellingShingle Banhong Liu
Mei Bai
Wuxiang Tu
Yanbin Shen
Jingxin Liu
Zhenquan Yang
Hongduo Bao
Qingli Dong
Yangtai Liu
Ran Wang
Hui Zhang
Liangbing Hu
The Virulence Factor LLO of <i>Listeria monocytogenes</i> Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect
Foods
<i>Listeria hemolysin</i>
biofilm
phage susceptibility
transcriptomics
title The Virulence Factor LLO of <i>Listeria monocytogenes</i> Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect
title_full The Virulence Factor LLO of <i>Listeria monocytogenes</i> Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect
title_fullStr The Virulence Factor LLO of <i>Listeria monocytogenes</i> Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect
title_full_unstemmed The Virulence Factor LLO of <i>Listeria monocytogenes</i> Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect
title_short The Virulence Factor LLO of <i>Listeria monocytogenes</i> Can Hamper Biofilm Formation and Indirectly Suppress Phage-Lytic Effect
title_sort virulence factor llo of i listeria monocytogenes i can hamper biofilm formation and indirectly suppress phage lytic effect
topic <i>Listeria hemolysin</i>
biofilm
phage susceptibility
transcriptomics
url https://www.mdpi.com/2304-8158/14/15/2554
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