Viable but non-culturable E. coli induced by low level chlorination have higher persistence to antibiotics than their culturable counterparts

Lin, Huirong; Ye, Chengsong; Chen, Sheng; Zhang, Shenghua; Yu, Xiubo

doi:10.1016/j.envpol.2017.06.047

Cited by 113 publications

(76 citation statements)

References 28 publications

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“…In fact, VBNC cells of Vibrio vulnificus have been shown to survive not only antibiotic treatment but also exposure to otherwise lethal levels of pH, heavy metals, heat, ethanol, and both osmotic and ionic challenges, establishing their propensity for stress resistance (59). Several studies have demonstrated the robust capability of VBNC cells of various species to survive antibiotics of multiple classes (15,26,36,(61)(62)(63)(64)(65)(66)(67). Similar to AP, several studies have demonstrated the presence of nonculturable cells at the clinical level (14,(68)(69)(70)(71)(72).…”

Section: Overlapping Phenotypic Characteristicsmentioning

confidence: 98%

Relationship between the Viable but Nonculturable State and Antibiotic Persister Cells

2018

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Bacteria have evolved numerous means of survival in adverse environments with dormancy, as represented by "persistence" and the "viable but nonculturable" (VBNC) state, now recognized to be common modes for such survival. VBNC cells have been defined as cells which, induced by some stress, become nonculturable on media that would normally support their growth but which can be demonstrated by various methods to be alive and capable of returning to a metabolically active and culturable state. Persister cells have been described as a population of cells which, while not being antibiotic resistant, are antibiotic tolerant. This drug-tolerant phenotype is thought to be a result of stress-induced and stochastic physiological changes as opposed to mutational events leading to true resistance. In this review, we describe these two dormancy strategies, characterize the molecular underpinnings of each state, and highlight the similarities and differences between them. We believe these survival modes represent a continuum between actively growing and dead cells, with VBNC cells being in a deeper state of dormancy than persister cells.

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Section: Overlapping Phenotypic Characteristicsmentioning

confidence: 98%

Relationship between the Viable but Nonculturable State and Antibiotic Persister Cells

2018

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“…Parent and adapted L. monocytogenes strains (inoculum: 2 × 10 8 cells mL −1 ) were incubated and gently shaken in 200 µL BHI without BC and in BHI supplemented with BC concentrations of 2, 3,4,5,6,8,9,10,11,12, and 13 µg mL −1 at 37 • C for 24 h. Colony forming units (CFUs) were determined in nine replicates according to the standard protocol of ISO 11290 [24].…”

Section: Determination Of Culturable Cells After Bc Exposurementioning

confidence: 99%

“…Adverse conditions may induce tolerance or resistance towards environmental stress factors and/or a shift in L. monocytogenes from a culturable state to a viable but nonculturable state (VBNC) [7]. VBNC cells are defined as bacterial cells that do not form colonies on standard culture media but still retain metabolic activity [8] and may revert to the active state [9]. Early VBNC studies coupled bacterial culture with microscopic counting Microorganisms 2020, 8, 184; doi:10.3390/microorganisms8020184 www.mdpi.com/journal/microorganisms of stained cells using SYTO 9/propidium iodide (SYTO9/PI), 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), 4 ,6-diamidino-2-phenylindole (DAPI), and/or fluorescently labelled antibodies [7,[10][11][12].…”

Section: Introductionmentioning

confidence: 99%

Benzalkonium Chloride Induces a VBNC State in Listeria monocytogenes

et al. 2020

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The objective of our study was to investigate the effects of benzalkonium chloride (BC) adaptation of L. monocytogenes on the susceptibility to antimicrobial agents and on the viable but non culturable (VBNC) state of the bacterial cells. We adapted L. monocytogenes SLCC2540 to BC by applying BC below minimum inhibitory concentration (MIC) to above minimum bactericidal concentration (MBC). The culturable fractions and the susceptibility of adapted and parental cells to BC were assessed. In addition, cell membrane permeability and glucose uptake were analyzed by multi parametric flow cytometry using the fluorescent agents SYTO9, propidium iodide, and 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-D-glucose (2-NBDG). Adapted cells displayed a two-fold MIC increase of BC and reduced antibiotic susceptibility. At high BC concentrations, the decrease in the number of colony forming units was significantly lower in the population of adapted cells compared to parental cells. At the same time, the number of metabolically active cells with intact membranes was significantly higher than the number of culturable cells. Growth-independent viability assays revealed an adapted subpopulation after BC application that was not culturable, indicating increased abundance of viable but nonculturable (VBNC) cells. Moreover, adapted cells can outcompete non-adapted cells under sublethal concentrations of disinfectants, which may lead to novel public health risks.

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“…In addition, Oliver et al reported that Vibrio vulnificus could enter the VBNC state, protecting cells against a variety of potentially lethal environmental challenges (heat, oxidative stress, osmotic stress, pH fluctuations, ethanol, antibiotics and heavy metals) 12 . In our previous study, it was found that low doses of chlorine (0.5 mg L −1 ) disinfection effectively reduced the number of culturable E. coli and induced a VBNC state, after which the metabolic activity of the bacteria was reduced and persistence of E. coli in the presence of 9 typical antibiotics was enhanced 13 . Moreover, Moreno et al also reported that low-level of residual chlorine in DWDS was capable of inducing VBNC state antibiotic-tolerant bacteria, which could be considered as an undetectable risk 8 .…”

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confidence: 95%

Characterization and potential mechanisms of highly antibiotic tolerant VBNC Escherichia coli induced by low level chlorination

Lin

Zhang

et al. 2020

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Escherichia coli is an important pathogenic indicator in drinking water. Viable but non-culturable (VBnc) E. coli induced by low level chlorination was found to have higher antibiotic tolerance. the emerging of VBnc bacteria in drinking water systems is posing challenges to the control of bio-safety. it is necessary to study the underlying mechanisms of VBnc state E. coli under actual residual chlorine condition of drinking water pipe network. In this study, we investigated the changes of morphology and gene expressions that might present such state. the results indicated that the size of VBnc E. coli was not remarkably changed or recovered culturability under favorable environmental conditions. Results from transcriptomic analysis revealed that the regulated genes related to fimbrial-like adhesin protein, putative periplasmic pilin chaperone, regulators of the transcriptional regulation, antibiotic resistance genes and stress-induced genes, rendering VBNC cells more tolerant to adverse environmental conditions. In total of 16 genes were significantly up-regulated under the VBNC state, including three genes encoding toxic protein (ygeG, ibsD, shoB), indicating that VBNC E. coil was still a threat to human. the work is of great relevance in the context of better understanding this poorly understood physiological state.

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Viable but non-culturable E. coli induced by low level chlorination have higher persistence to antibiotics than their culturable counterparts

Cited by 113 publications

References 28 publications

Relationship between the Viable but Nonculturable State and Antibiotic Persister Cells

Relationship between the Viable but Nonculturable State and Antibiotic Persister Cells

Benzalkonium Chloride Induces a VBNC State in Listeria monocytogenes

Characterization and potential mechanisms of highly antibiotic tolerant VBNC Escherichia coli induced by low level chlorination

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