VirR-Mediated Resistance of Listeria monocytogenes against Food Antimicrobials and Cross-Protection Induced by Exposure to Organic Acid Salts.
Formulations of ready-to-eat (RTE) foods with antimicrobial compounds constitute an important safety measure against foodborne pathogens such as Listeria monocytogenes. While the efficacy of many commercially available antimicrobial compounds has been demonstrated in a variety of foods, the current understanding of the resistance mechanisms employed by L. monocytogenes to counteract these stresses is limited. In this study, we screened in-frame deletion mutants of two-component system response regulators associated with the cell envelope stress response for increased sensitivity to commercially available antimicrobial compounds (nisin, lauric arginate, ε-polylysine, and chitosan). A virR deletion mutant showed increased sensitivity to all antimicrobials and significantly greater loss of membrane integrity when exposed to nisin, lauric arginate, or ε-polylysine (P < 0.05). The VirR-regulated operon, dltABCD, was shown to be the key contributor to resistance against these antimicrobial compounds, whereas another VirR-regulated gene, mprF, displayed an antimicrobial-specific contribution to resistance. An experiment with a β-glucuronidase (GUS) reporter fusion with the dlt promoter indicated that nisin does not specifically induce VirR-dependent upregulation of dltABCD. Lastly, prior exposure of L. monocytogenes parent strain H7858 and the ΔvirR mutant to 2% potassium lactate enhanced subsequent resistance against nisin and ε-polylysine (P < 0.05). These data demonstrate that VirRS-mediated regulation of dltABCD is the major resistance mechanism used by L. monocytogenes against cell envelope-damaging food antimicrobials. Further, the potential for cross-protection induced by other food-related stresses (e.g., organic acids) needs to be considered when applying these novel food antimicrobials as a hurdle strategy for RTE foods.
Department of Food Science, Cornell University, Ithaca, New York, USA.
Jihun
J
Kang
Department of Food Science, Cornell University, Ithaca, New York, USA.
Martin
M
Wiedmann
Department of Food Science, Cornell University, Ithaca, New York, USA.
Kathryn J
KJ
Boor
Department of Food Science, Cornell University, Ithaca, New York, USA Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo, North Dakota, USA Teresa.bergholz@ndsu.edu.
Teresa M
TM
Bergholz
D000900:Anti-Bacterial Agents / Q000633:toxicity*; D001120:Arginine / Q000031:analogs & derivatives / Q000633:toxicity; D024881:Drug Resistance, Bacterial; D005520:Food Preservatives / Q000633:toxicity*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005820:Genetic Testing; D008089:Listeria monocytogenes / Q000187:drug effects*; D009561:Nisin / Q000633:toxicity; D009876:Operon; D011107:Polylysine / Q000633:toxicity; D014157:Transcription Factors / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D005520:Food Preservatives; D014157:Transcription Factors; D009561:Nisin; D011107:Polylysine; C031371:N(alpha)-lauroylarginine ethyl ester; D001120:Arginine