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Viable but nonculturable (VBNC) Vibrio parahaemolyticus cannot be detected by the standard cultivation-based methods. In this study, commonly used viability assessment methods were evaluated for the detection of Vibrio parahaemolyticus in a VBNC state. V. parahaemolyticus cells exposed to nutrient-deficiency at cold temperature were used for epifluorescence microscopy with SYTO and propidium iodide (PI) staining and real-time polymerase chain reaction (qPCR) with propidium monoazide (PMA), and its resuscitative ability was determined by a temperature upshift in freshly prepared artificial sea water (ASW; pH 7) fluids. Viable cells with intact membranes always exceeded 5.0 log CFU ml in ASW microcosms at 4°C. After 80 days, cycle thresholds for V. parahaemolyticus ATCC 27969 were 16.15-16.69. During cold-starvation, PMA qPCR selectively excluded DNAs from heat-killed cells. However, there may be some penetration of PMA into undamaged cells that persisted in ASW for 150 days, as evidenced by their ability to resuscitate from a VBNC state after a temperature upshift (25°C); V. parahaemolyticus ATCC 33844 and V. parahaemolyticus ATCC 27969 were successfully reactivated from a VBNC state in ASW microcosms containing <5% NaCl, following enrichment in ASW medium (pH 7). This article is protected by copyright. All rights reserved.
This article was published in the following journal.
Name: Letters in applied microbiology
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A species of bacteria found in the marine environment, sea foods, and the feces of patients with acute enteritis.
A species of gram-negative bacteria in the genus VIBRIO, the causative agent of cold water vibriosis, a HEMORRHAGIC SEPTICEMIA of ATLANTIC SALMON.
A condition of the feet produced by prolonged exposure of the feet to water. Exposure for 48 hours or more to warm water causes tropical immersion foot or warm-water immersion foot common in Vietnam where troops were exposed to prolonged or repeated wading in paddy fields or streams. Trench foot results from prolonged exposure to cold, without actual freezing. It was common in trench warfare during World War I, when soldiers stood, sometimes for hours, in trenches with a few inches of cold water in them. (Andrews' Diseases of the Skin, 8th ed, p27)
Cellular proteins and peptides that are induced in response to cold stress. They are found in a broad variety of prokaryotic and eukaryotic organisms.
A strain of the VIBRIO CHOLERAE bacteria belonging to serogroup non-O1, infecting humans and other PRIMATES. It is related to VIBRIO CHOLERAE O1, but causes a disease less severe than CHOLERA. Eating raw shellfish contaminated with the bacteria results in GASTROENTERITIS.
Polymerase Chain Reaction (PCR)
PCR (Polymerase Chain Reaction) uses the ability of DNA polymerase (enzymes that create DNA molecules by assembling nucleotides, the building blocks of DNA. These enzymes are essential to DNA replication and usually work in pairs to create two ident...