Induction of <i>Escherichia coli</i> O157:H7 into the viable but non‐culturable state by chloraminated water and river water, and subsequent resuscitation

Liu, Yanming; Wang, Chuan; Tyrrell, Gregory J.; Hrudey, Steve E.; Li, Xing‐Fang

doi:10.1111/j.1758-2229.2009.00024.x

Cited by 50 publications

(45 citation statements)

References 24 publications

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“…that antibiotic-resistant strains of S. enterica serovar Newport persisted for longer periods of time than their nonresistant counterparts in manure-amended soils. A fourth explanation may be that organisms enter a viable but not cultivable (VBNC) state, resulting in the appearance of an initial rapid decay, until stressed cells alter their physiology, followed by slower decay of more resilient organisms that remain cultivable, or as cells transition into and out of the VBNC state (8,12,14,27). Other potential sources of this behavior may include predation, biofilm formation, or the acquisition of genetic materials by a subpopulation of fecal bacteria from the native soil microbial community that favor their survival over counterparts that do not similarly adapt.…”

Section: Discussionmentioning

confidence: 99%

Decay of Bacterial Pathogens, Fecal Indicators, and Real-Time Quantitative PCR Genetic Markers in Manure-Amended Soils

Rogers

Donnelly

Peed

et al. 2011

Appl Environ Microbiol

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This study examined persistence and decay of bacterial pathogens, fecal indicator bacteria (FIB), and emerging real-time quantitative PCR (qPCR) genetic markers for rapid detection of fecal pollution in manureamended agricultural soils. Known concentrations of transformed green fluorescent protein-expressing Escherichia coli O157:H7/pZs and red fluorescent protein-expressing Salmonella enterica serovar Typhimurium/pDs were added to laboratory-scale manure-amended soil microcosms with moisture contents of 60% or 80% field capacity and incubated at temperatures of ؊20°C, 10°C, or 25°C for 120 days. A two-stage first-order decay model was used to determine stage 1 and stage 2 first-order decay rate coefficients and transition times for each organism and qPCR genetic marker in each treatment. Genetic markers for FIB (Enterococcus spp., E. coli, and Bacteroidales) exhibited decay rate coefficients similar to that of E. coli O157:H7/pZs but not of S. enterica serovar Typhimurium/pDs and persisted at detectable levels longer than both pathogens. Concentrations of these two bacterial pathogens, their counterpart qPCR genetic markers (stx1 and ttrRSBCA, respectively), and FIB genetic markers were also correlated (r ‫؍‬ 0.528 to 0.745). This suggests that these qPCR genetic markers may be reliable conservative surrogates for monitoring fecal pollution from manure-amended land. Hostassociated qPCR genetic markers for microbial source tracking decayed rapidly to nondetectable concentrations, long before FIB, Salmonella enterica serovar Typhimurium/pDs, and E. coli O157:H7/pZs. Although good indicators of point source or recent nonpoint source fecal contamination events, these host-associated qPCR genetic markers may not be reliable indicators of nonpoint source fecal contamination events that occur weeks following manure application on land.Cultivation-based methods for fecal indicator bacteria (FIB) such as Escherichia coli and Enterococcus spp. have long been used to indicate potential public health risks associated with water impacted by human and other animal feces (53). FIB cultivation methods are simple to perform and inexpensive. However, these methods require 18 to 24 h following sampling to generate test results; this allows potential exposure of the public to fecal pathogens in the interim. Regulatory agencies, business owners, and other stakeholders have expressed interest in more rapid and specific methods to identify water quality impairment.Emerging real-time quantitative PCR (qPCR) methods designed to estimate the concentration of fecal pollution by targeting genomic DNA (gDNA) from FIB such as Bacteroidales, Enterococcus spp., and E. coli are now available and can generate test results in just a few hours after sampling (10,16,48). Some of these genetic markers can be correlated to public health risk and may soon be incorporated by the U.S. Environmental Protection Agency into water quality standards in the United States (16,59). These genetic markers may also detect viable but nonculturable (VBNC) cells tha...

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Section: Discussionmentioning

confidence: 99%

Decay of Bacterial Pathogens, Fecal Indicators, and Real-Time Quantitative PCR Genetic Markers in Manure-Amended Soils

Rogers

Donnelly

Peed

et al. 2011

Appl Environ Microbiol

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“…Further, it was seen that the use of “conditioned” media (supernatants of the culture media in which E. coli cells were grown previously), prolonged the duration of the VBNC state. The viability of E. coli O157:H7 10 months after its induction into the VBNC state by river or chlorinated waters has been demonstrated by establishing the presence of mRNAs of rfbE and fliC genes that encode parts of O-antigen transporter and flagellar filament structural proteins, respectively (21). …”

Section: Proteomics and Transcriptomics Of Vbnc Cellsmentioning

confidence: 99%

“…Some of the human pathogens, such as E. coli O157:H7, can be resuscitated using its autoinducers such as 2(A1-2) produced during biofilm formation in a serum-based medium (21). Combinations of amino acids such as methionine, glutamine, threonine, serine, and asparagine in the basal minimal medium are also known to support the resuscitation of E. coli cells (44).…”

Section: Resuscitation Of Vbnc Bacteriamentioning

confidence: 99%

Current Perspectives on Viable but Non-Culturable (VBNC) Pathogenic Bacteria

Ramamurthy

Ghosh

Pazhani

et al. 2014

Front. Public Health

357

254

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Under stress conditions, many species of bacteria enter into starvation mode of metabolism or a physiologically viable but non-culturable (VBNC) state. Several human pathogenic bacteria have been reported to enter into the VBNC state under these conditions. The pathogenic VBNC bacteria cannot be grown using conventional culture media, although they continue to retain their viability and express their virulence. Though there have been debates on the VBNC concept in the past, several molecular studies have shown that not only can the VBNC state be induced under in vitro conditions but also that resuscitation from this state is possible under appropriate conditions. The most notable advance in resuscitating VBNC bacteria is the discovery of resuscitation-promoting factor (Rpf), which is a bacterial cytokines found in both Gram-positive and Gram-negative organisms. VBNC state is a survival strategy adopted by the bacteria, which has important implication in several fields, including environmental monitoring, food technology, and infectious disease management; and hence it is important to investigate the association of bacterial pathogens under VBNC state and the water/foodborne outbreaks. In this review, we describe various aspects of VBNC bacteria, which include their proteomic and genetic profiles under the VBNC state, conditions of resuscitation, methods of detection, antibiotic resistance, and observations on Rpf.

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“…In general, this storage method has been considered an effective means of reducing the possibility of infection, however, increasing demand for long-life fresh products creates new risk such as viable but non-culturable (VBNC) bacteria in the cold chain. A number of pathogens (e.g., Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Oranienburg, Vibrio cholerae, Helicobacter pylori, Listeria monocytogenes) enter a VBNC state in response to various environmental stresses such as cold exposure, pH, and high osmolarity [2,3,8,34,40]. In a VBNC state, the bacteria cannot be detected by conventional culture-based methods due to the loss of culturability on the media that normally support their growth, but remain viable and express various degrees of metabolic activities [12,16,37].…”

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confidence: 99%

Physiological Characterization of Campylobacter jejuni under Cold Stresses Conditions: Its Potential for Public Threat

et al. 2012

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ABSTRACT. Campylobacter jejuni is the major cause of human gastroenteritis worldwide. Under stress conditions, C. jejuni can enter a viable but non-culturable (VBNC) state. We found that the C. jejuni was able to enter a VBNC state by prolonged incubation at 4C. The standard isolation methods using pre-enrichment steps in Bolton broth or Preston broth could not detect the VBNC cells in spiked chicken meat. The transcription levels of virulence-associated genes (flaA, flaB, cadF, ciaB, cdtA, cdtB and cdtC) were expressed in VBNC cells but in low levels. The VBNC cells retained the ability to invade Caco-2 human intestinal epithelial cells in vitro. In most cases, VBNC cells failed to resuscitate in Caco-2 cells, but in some experiments, they formed colonies after co-incubation with host cells. Collectively, C. jejuni enters into a VBNC state at 4C and the VBNC C. jejuni remains virulent which may possibly lead to disease in humans. C. jejuni in VBNC state is a potential concern for food safety.KEY WORDS: Campylobacter jejuni, resuscitation, viable but nonculturable state, virulence.

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Induction of Escherichia coli O157:H7 into the viable but non‐culturable state by chloraminated water and river water, and subsequent resuscitation

Cited by 50 publications

References 24 publications

Decay of Bacterial Pathogens, Fecal Indicators, and Real-Time Quantitative PCR Genetic Markers in Manure-Amended Soils

Decay of Bacterial Pathogens, Fecal Indicators, and Real-Time Quantitative PCR Genetic Markers in Manure-Amended Soils

Current Perspectives on Viable but Non-Culturable (VBNC) Pathogenic Bacteria

Physiological Characterization of Campylobacter jejuni under Cold Stresses Conditions: Its Potential for Public Threat

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