Proteomic Characterization of Enterohemorrhagic <i>Escherichia coli</i> O157:H7 in the Oxidation‐Induced Viable but Non‐Culturable State

Asakura, Hiroshi; Panutdaporn, Nantika; Kawamoto, Keiko; Igimi, Shizunobu; Yamamoto, Shigeki; Makino, Sou‐ichi

doi:10.1111/j.1348-0421.2007.tb03969.x

Cited by 44 publications

(31 citation statements)

References 23 publications

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“…The constitutive and stable expression of the virulence genes hly and stx 1 and stx 2 and enterotoxin production in VBNC cells have been reported (5,24,32,40). In our work we recorded the presence of verotoxin for 3 days in samples that contained a VBNC population of approximately 4.5 log 10 bacteria and no detectable culturable cells.…”

Section: Discussionmentioning

confidence: 79%

“…The possibility that the VBNC state of E. coli O157:H7 contributed to a food-borne outbreak was suggested by Makino et al in a study on strains isolated from salted salmon roe (25). Moreover, the expression of the Shiga-toxin genes in VBNC cells (5,24,40) and resuscitation by bacterial growth inducer, e.g., H 2 O 2 -degrading compounds such as sodium pyruvate and the antioxidant Oxyrase (4,33), have been demonstrated. Also, pathogenic bacteria linked with the consumption of the fresh produce, including Salmonella spp.…”

mentioning

confidence: 97%

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Induction of Viable but Nonculturable Escherichia coli O157:H7 in the Phyllosphere of Lettuce: a Food Safety Risk Factor

Dinu

Bach

2011

Appl Environ Microbiol

129

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Escherichia coli O157:H7 continues to be an important human pathogen and has been increasingly linked to food-borne illness associated with fresh produce, particularly leafy greens. The aim of this work was to investigate the fate of E. coli O157:H7 on the phyllosphere of lettuce under low temperature and to evaluate the potential hazard of viable but nonculturable (VBNC) cells induced under such stressful conditions. First, we studied the survival of six bacterial strains following prolonged storage in water at low temperature (4°C) and selected two strains with different nonculturable responses for the construction of E. coli O157:H7 Tn7gfp transformants in order to quantitatively assess the occurrence of human pathogens on the plant surface. Under a suboptimal growth temperature (16°C), both E. coli O157:H7 strains maintained culturability on lettuce leaves, but under more stressful conditions (8°C), the bacterial populations evolved toward the VBNC state. The strain-dependent nonculturable response was more evident in the experiments with different inoculum doses (10 9 and 10 6 E. coli O157:H7 bacteria per g of leaf) when strain BRMSID 188 lost culturability after 15 days and strain ATCC 43895 lost culturability within 7 days, regardless of the inoculum dose. However, the number of cells entering the VBNC state in high-cell-density inoculum (approximately 55%) was lower than in low-cell-density inoculum (approximately 70%). We recorded the presence of verotoxin for 3 days in samples that contained a VBNC population of 4 to 5 log 10 cells but did not detect culturable cells. These findings indicate that E. coli O157:H7 VBNC cells are induced on lettuce plants, and this may have implications regarding food safety.

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Section: Discussionmentioning

confidence: 79%

mentioning

confidence: 97%

Induction of Viable but Nonculturable Escherichia coli O157:H7 in the Phyllosphere of Lettuce: a Food Safety Risk Factor

Dinu

Bach

2011

Appl Environ Microbiol

129

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“…Using molecular techniques, gene transcription and the synthesis and/or modulation of proteins in cells in the VBNC state have been demonstrated (Asakura et al 2007(Asakura et al , 2008Coutard et al 2005;Fischer-Le Saux et al 2002;Gunasekera et al 2002;Heim et al 2002;Karunasagar and Karunasagar 2005;Lleó et al 2000;Smith and Oliver 2006). These facts ratify unequivocally that populations in the VBNC state are formed by live cells.…”

Section: The Viable But Nonculturable Statementioning

confidence: 76%

“…Xu et al (1982) defined this phenomenon as the viable but nonculturable (VBNC) state. Both the influence of environmental factors in the dynamics of the formation of this phenotype, as well as the morphological characteristics and the physiological properties of cells in the VBNC state, including gene expression and the modulation of novel proteins, have been widely studied Asakura et al 2007Asakura et al , 2008; Barcina et al 1997;Coutard et al 2007;Cuny et al 2005;Day and Oliver 2004;Lleó et al 2000;Muela et al 1999Muela et al , 2008Smith and Oliver 2006;Srinivasan et al 1998;Wong et al 2004a). …”

Section: The Viable But Nonculturable Statementioning

confidence: 97%

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The viable but nonculturable phenotype: a crossroads in the life-cycle of non-differentiating bacteria?

Barcina

Arana

2009

Rev Environ Sci Biotechnol

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In nature, prokaryotes must face alternating periods of prosperity and adversity. Differentiating bacteria confront situations of adversity by developing resistant structures. When there is a plentiful period, they adopt a vegetative state and when the period is adverse, a resistant structure, thereby completing a cycle. Non-differentiating bacteria do not develop such morphological distinct resistant structures. It has been proposed that many of these bacteria withstand periods of adversity by adopting the viable but nonculturable phenotype (VBNC). Bacteria of this phenotype conserve detectable metabolic function but become unculturable. Is it possible that the VBNC phenotype can revert to a culturable state, and vice versa, thus establishing a life-cycle? This review presents and evaluates different hypotheses regarding this question. Moreover, it attempts to analyse and proffer answers to other questions related to this phenotype. Is this a successful phenotype which prolongs survival? Is this a strategy for the survival of individual cells, or is it a strategy for the survival of a population? Finally, is it possible that this phenotype is, in fact, an example of altruistic death?

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Systems Biology: Integrating ‘‐Omics'‐Oriented Approaches to Determine Foodborne Microbial Toxins

Singh¹,

Nagaraj²,

Gabani³

2009

General, Applied and Systems Toxicology

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The possible origins of microbial toxins vary widely, and detection of these toxins in different food matrices is a major challenge for food industries and regulatory agencies. New methodologies are needed to quickly and precisely detect traces of micro‐organisms and their toxic metabolites. In post‐genomics era, systems biology approaches, ranging from genomic sequencing to transcriptomics, proteomics and metabolomic profiling, may be an effective platform for developing tests to identify a variety of toxins in field applications; multiple functional ‘‐omics’ could be combined into a system‐wide approach for detecting toxins, which may also be useful in the study of microbial pathogenesis. Advances in systems biology are addressed in the current article, as well as possible uses of these high‐throughput platforms to ensure food and feed safety.

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Proteomic Characterization of Enterohemorrhagic Escherichia coli O157:H7 in the Oxidation‐Induced Viable but Non‐Culturable State

Cited by 44 publications

References 23 publications

Induction of Viable but Nonculturable Escherichia coli O157:H7 in the Phyllosphere of Lettuce: a Food Safety Risk Factor

Induction of Viable but Nonculturable Escherichia coli O157:H7 in the Phyllosphere of Lettuce: a Food Safety Risk Factor

The viable but nonculturable phenotype: a crossroads in the life-cycle of non-differentiating bacteria?

Systems Biology: Integrating ‘‐Omics'‐Oriented Approaches to Determine Foodborne Microbial Toxins

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