The Public Health Significance of Viable but Nonculturable Bacteria

Oliver, James D.

doi:10.1007/978-1-4757-0271-2_16

Cited by 143 publications

(138 citation statements)

References 83 publications

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“…FCM is a quantitative technique and allows an easy measurement of size variations in the total number of cells present in a population, whereas SEM images permits us to observe morphological changes. Our data are in accord with some authors (10,11) who noted that during the entry into the VBNC state, bacteria become smaller in size, acquiring coccal morphology, even when key activities of the cellular metabolism were still detectable (50).…”

Section: Vparahaemolyticus Vbnc State By Flow Cytometrysupporting

confidence: 93%

“…In this state, although still expressing various degrees of metabolic activity and possibly being able to cause infection (6,8,9), the bacterial cells cannot be cultured on conventional laboratory media that normally support their growth. During entry into the VBNC state, bacteria acquire a coccal morphology, become smaller in size (10,11), and show changes in their protein (12,13) and fatty acid profiles (14). The nucleic acid content, in particular RNA, can vary when bacteria enter the VBNC state, justifying the reduced RNA levels which result from nutritional downshifts in bacteria (15).…”

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confidence: 99%

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State transitions of Vibrio parahaemolyticus VBNC cells evaluated by flow cytometry

Falcioni

Papa

Campana

et al. 2008

Cytometry Part B Clinical

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Background: Vibrio parahaemolyticus, in response to environmental conditions, may be present in a viable but nonculturable state (VBNC), which can still be responsible for cases of infectious diseases in humans.Methods: The characterization of the cellular states of V. parahaemolyticus during entry into, persistence in, and resuscitation from the VBNC state, was assessed through plate culture method and epifluorescence microscope evaluation of actively respiring cells. Flow cytometry (FCM) in combination with SYBR Green I (SG) and propidium iodide allowed us to distinguish between viable, dead, and damagedcells. Immunofluorescence labeling detected by FCM was used to study changes in antibody affinity.Results: Two groups of bacteria, one with High Nucleic Acid (HNA) and one having Low Nucleic Acid (LNA) content, were differentiated using SG and FCM and each was correlated with cell viability. With the aging of the microcosm, the LNA bacteria population increased while the HNA population gradually disappeared. Cytofluorimetric immunofluorescence analyses showed that the bacterial cell levels dropped from 95% at day 0 to 40% at day 26 and by day 29, antibody affinity was virtually lost. FCM analyses of light scatter signals expressed by cell population highlighted morphological changes indicating a reduction in cell size, as also shown by scanning electron microscopy images and variations in cell structure.Conclusions: The methodology used has provided useful data in relation to the state transitions of V. parahaemolyticus regarding cell viability, antigenic surface components, and the quantification of morphological variations during its entry into the VBNC state. q

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Section: Vparahaemolyticus Vbnc State By Flow Cytometrysupporting

confidence: 93%

mentioning

confidence: 99%

State transitions of Vibrio parahaemolyticus VBNC cells evaluated by flow cytometry

Falcioni

Papa

Campana

et al. 2008

Cytometry Part B Clinical

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“…have also been shown to induce several bacteria, including E. coli and S. typhimurium, into a viable but nonculturable (VBNC) state (Oliver 1993(Oliver , 2000a. Cells in the VBNC state are viable, as can be demonstrated by several direct microscopic assays (Oliver 1993(Oliver , 2000bBreeuwer & Abee 2000;Cré ach et al 2003) but lose the ability to be cultured on routine microbiological media.…”

Section: Mcfeters and Lechevallier 2000) Such Environmental Stressesmentioning

confidence: 99%

“…Cells in the VBNC state are viable, as can be demonstrated by several direct microscopic assays (Oliver 1993(Oliver , 2000bBreeuwer & Abee 2000;Cré ach et al 2003) but lose the ability to be cultured on routine microbiological media. This state is characterized by significant modifications in cell morphology, metabolism, physiology and possibly genomic structure (Oliver 2000a,b (Oliver & Bockian 1995;Colwell et al 1996;Pommepuy et al 1996;Oliver 2000a). Thus, chlorination of wastewater might be assumed to be disinfecting when in fact some of the cells that were supposedly inactivated may remain viable, resuscitate down the line, possibly increase in number, and cause infection.…”

Section: Mcfeters and Lechevallier 2000) Such Environmental Stressesmentioning

confidence: 99%

Induction of Escherichia coli and Salmonella typhimurium into the viable but nonculturable state following chlorination of wastewater

Oliver

Dagher

Linden

2005

Journal of Water and Health

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128

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We examined the effects of chlorine disinfection on Escherichia coli and Salmonella typhimurium in secondary-treated wastewater to determine whether such treatment might induce these bacteria into the viable but nonculturable (VBNC) state. In this state, cells lose culturability but retain viability and the potential to revert to the metabolically active and infectious state.To examine the effects of chlorination on cells in different physiological states, cells from the logarithmic and stationary phases, or nutrient starved, or grown in natural wastewater, were studied. Isogenic cells with and without plasmids were also examined. Whereas a mixture of free and combined chlorine, as occurs under typical wastewater disinfection, was found to be rapidly lethal to most cells, regardless of their physiological state or plasmid content, c. 10 4 of the original 10 6 cells ml 21 did survive in the VBNC state. While we were not successful in resuscitating these cells to the culturable state, the presence of such nonculturable cells in treated wastewater offers a potential public health hazard.

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“…For these reasons it is difficult to correlate the two approaches although this is commonly done due to a lack of a practical alternative standard reference method [9,12,16]. There is a growing body of evidence pointing to the virulence of VBNC bacteria and suggesting the importance of measuring their numbers in environmental waters [17][18][19]. For this purpose enzyme assays are a measurement technology which could be implemented and optimised.…”

Section: Introductionmentioning

confidence: 99%

ColiSense, today's sample today: A rapid on-site detection of β-d-Glucuronidase activity in surface water as a surrogate for E. coli

Heery

Briciu-Burghina

Zhang

et al. 2016

Talanta

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ab s t r a c t A sensitive field-portable fluorimeter with incubating capability and triplicate sample chambers was designed and built. The system was optimised for the on-site analysis of E. coli in recreational waters using fluorescent based enzyme assays. The target analyte was β-D-Glucuronidase (GUS) which hydrolyses a synthetic substrate 6-Chloro-4-Methyl-Umbelliferyl-β-D-Glucuronide (6-CMUG) to release the fluorescent molecule 6-Chloro-4-Methyl-Umbelliferyl (6-CMU). The system was calibrated with 6-CMU standards. A LOD of 5 nM and a resolution of less than 1 nM was determined while enzyme kinetic tests showed detection of activities below 1 pmol min 1 mL 1 of sample. A field portable sample preparation, enzyme extraction protocol and continuous assay were applied with the system to analyse freshwater and marine samples. Results from a one day field trial are shown which demonstrated the ability of the system to deliver results on-site within a 75 min period.

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The Public Health Significance of Viable but Nonculturable Bacteria

Cited by 143 publications

References 83 publications

State transitions of Vibrio parahaemolyticus VBNC cells evaluated by flow cytometry

State transitions of Vibrio parahaemolyticus VBNC cells evaluated by flow cytometry

Induction of Escherichia coli and Salmonella typhimurium into the viable but nonculturable state following chlorination of wastewater

ColiSense, today's sample today: A rapid on-site detection of β-d-Glucuronidase activity in surface water as a surrogate for E. coli

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