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The detection, enumeration, and virulence potential of viable but non-culturable (VBNC) pathogens continues to be a topic of discussion. While there is a lack of definitive evidence that VBNC (Lm) pose a public health risk, recent studies suggest that Lm in its VBNC state remains virulent. VBNC bacteria cannot be enumerated by traditional plating methods, so the results from routine Lm testing may not demonstrate a sample's true hazard to public health. We suggest that supplementing routine Lm testing methods with methods designed to enumerate VBNC cells may more accurately represent the true level of risk. This review summarizes five methods for enumerating VNBC Lm: Live/Dead BacLight staining, ethidium monoazide and propidium monoazide-stained real-time polymerase chain reaction (EMA- and PMA-PCR), direct viable count (DVC), 5-cyano-2,3-ditolyl tetrazolium chloride-4',6-diamidino-2-phenylindole (CTC-DAPI) double staining, and carboxy-fluorescein diacetate (CDFA) staining. Of these five supplementary methods, the Live/Dead BacLight staining and CFDA-DVC staining currently appear to be the most accurate for VBNC Lm enumeration. In addition, the impact of the VBNC state on the virulence of Lm is reviewed. Widespread use of these supplemental methods would provide supporting data to identify the conditions under which Lm can revert from its VBNC state into an actively multiplying state and help identify the environmental triggers that can cause Lm to become virulent. Highlights: Rationale for testing for all viable Listeria (Lm) is presented. Routine environmental sampling and plating methods may miss viable Lm cells. An overview and comparison of available VBNC testing methods is given. There is a need for resuscitation techniques to recover Lm from VBNC. A review of testing results for post VBNC virulence is compared.
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http://dx.doi.org/10.3390/microorganisms9010194 | DOI Listing |
Infect Immun
December 2024
Laboratory of Applied Immunology, Institute of Biology Sciences, University of Brasília, Brasília, Brazil.
Dormancy is an adaptation in which cells reduce their metabolism, transcription, and translation to stay alive under stressful conditions, preserving the capacity to reactivate once the environment reverts to favorable conditions. Dormancy and reactivation of () are closely linked to intracellular residency within macrophages. Our previous work showed that murine macrophages rely on the viable but not cultivable (VBNC-a dormancy phenotype) fungus from active , with striking differences in immunometabolic gene expression.
View Article and Find Full Text PDFJ Food Prot
December 2024
Department of Biosystems and Agricultural Engineering, Michigan State University, East Lansing, MI 48824, USA. Electronic address:
Rapid detection of bacterial pathogens is essential for food safety and public health, yet bacteria can evade detection by entering a viable but nonculturable (VBNC) state under sublethal stress, such as antimicrobial residues. These bacteria remain active but undetectable by standard culture-based methods without extensive enrichment, necessitating advanced detection methods. This study developed an AI-enabled hyperspectral microscope imaging (HMI) framework for rapid VBNC detection under low-level antimicrobials.
View Article and Find Full Text PDFFront Cell Infect Microbiol
December 2024
Key Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou, China.
The zoonotic pathogen is responsible for diverse human diseases, from mild to life-threatening, but it often eludes detection in culture-based assays. This study investigates the potential of to enter a viable but nonculturable (VBNC) state when exposed to human fever temperature or antibiotics, with this state confirmed by successful resuscitation. Viability was assessed using SYBR Green I/PI staining and propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR), while culturability was determined through colony-forming unit (CFU) counting on blood agar plates.
View Article and Find Full Text PDFJ Environ Sci (China)
June 2025
School of Environmental Science and Engineering, Laboratory of Marine Ecological Environment in Universities of Shandong, Shandong University, Qingdao 266237, China; Qingdao Key Laboratory of Marine Pollutant Prevention, Shandong Key Laboratory of Environmental Processes and Health, Shandong University, Qingdao 266237, China. Electronic address:
Advanced oxidation processes (AOPs) exhibit significant potential for water disinfection due to their generation of large quantities of highly oxidizing free radicals. However, the neglect of viable but nonculturable (VBNC) cells obscures their true disinfection efficacy and potential environmental health risks. Therefore, the study evaluated the disinfection effectiveness and mechanisms of typical AOPs, including Fe/HO, Fe/persulfate (PS), and O, from the perspective of the production of VBNC bacteria.
View Article and Find Full Text PDFMicroorganisms
November 2024
Departamento de Microbiología Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (ENCB, IPN), Ciudad de México 11340, Mexico.
The general features of the shift to a dormant state in mycobacterial species include several phenotypic changes, reduced metabolic activities, and increased resistance to host and environmental stress conditions. In this study, we aimed to provide novel insights into the viability state and morphological changes in dormant that contribute to its long-term survival under starvation or hypoxia. To this end, we conducted assays to evaluate cell viability, morphological changes and gene expression.
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