A comparison of solid and liquid media for resuscitation of starvation- and low-temperature-induced nonculturable cells of Aeromonas hydrophila

Wai, Sun Nyunt; Mizunoe, Yoshimitsu; Takade, Akemi; Yoshida, Shigeaki

doi:10.1007/s002030000142

Cited by 89 publications

(69 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Sodium pyruvate and H 2 O 2 -degrading agents seem to be effective compounds for recovering the physiological state after cell damage or a loss of culturability (1,12,46,48,49). In our study, amendment (75). This effect may be explained by the fact that 20°C is closer to the optimal growth temperature of the bacteria.…”

Section: Discussionmentioning

confidence: 60%

“…Rahman et al (60) reported that A. hydrophila cultured in a 0.35% NaCl solution at pH 7.5 and 25°C for 50 days showed a VBNC state characterized by the presence of viable but nonculturable bacteria. In our study, the culturability of the aroA strain declined more rapidly at 6°C than at the higher temperatures, and it has been reported that low temperatures seem to favor entry into the VBNC or ANC state (21,75). To clarify this point, we tested a method for in vitro resuscitation, studied the morphological changes, and assessed the viability of the nonculturable A. hydrophila cells by using an indirect labeling method to distinguish cells with damaged and intact cell membranes.…”

Section: Discussionmentioning

confidence: 70%

“…During the experiments, samples from the various microcosms were serially diluted and used to inoculate TSA amended with sodium pyruvate (0.1% [wt/vol]) as described by Mizunoe et al (48) and Wai et al (75). Sodium pyruvate was added directly to TSA prior to autoclaving.…”

Section: Methodsmentioning

confidence: 99%

“…Under adverse conditions, such as those present in some natural aquatic environments, A. hydrophila strains are able to enter a viable but nonculturable (VBNC) or an active nonculturable (ANC) state (40,44,75), from which they may be resuscitated under permissive conditions (40,75). A VBNC or an ANC state allows microorganisms long-term permanence in the environment and makes their isolation for diagnostic procedures difficult (9,31).…”

Section: Cfu MLmentioning

confidence: 99%

See 3 more Smart Citations

Behavior of an Aeromonas hydrophila aroA Live Vaccine in Water Microcosms

Vivas¹,

Carracedo²,

Riaño³

et al. 2004

Appl Environ Microbiol

View full text Add to dashboard Cite

Genetically modified auxotrophic mutants of different fish pathogens have been used as live vaccines in laboratory experiments, but the behavior of the strains after release into aquatic ecosystems has not been characterized. We previously constructed and characterized an aroA mutant of Aeromonas hydrophila and studied the protection afforded by this mutant as a live vaccine in rainbow trout. In this work, we describe the survival of this strain in aquatic microcosms prepared from fish water tanks. The aroA mutant disappeared rapidly in nonfiltered, nonautoclaved fish tank water, declining below detection levels after 15 days, suggesting an inhibitory effect of the autochthonous microflora of the water. When the aroA strain was used to inoculate sterilized water, its culturability was lower than that of wild-type strain A. hydrophila AG2; after long periods of incubation, aroA cells were able to enter a viable but nonculturable state. Entry into this nonculturable state was accompanied by changes in the cell morphology from rods to spheres, but the cells appeared to remain potentially viable, as assessed by the preservation of cell membrane integrity. Supplementation of the culture medium with sodium pyruvate favored the culturability and resuscitation of the two A. hydrophila strains at low temperatures (6 and 16°C). These results contribute to a better understanding of the behavior of the aroA strain in natural environments and suggest that the inactivation of the aroA gene may be beneficial for the safety of this live vaccine for aquacultures.

show abstract

Section: Discussionmentioning

confidence: 60%

Section: Discussionmentioning

confidence: 70%

Section: Methodsmentioning

confidence: 99%

Section: Cfu MLmentioning

confidence: 99%

See 2 more Smart Citations

Behavior of an Aeromonas hydrophila aroA Live Vaccine in Water Microcosms

Vivas¹,

Carracedo²,

Riaño³

et al. 2004

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…However, more recent results indicated that V. vulnificus cells previously considered as VBNC actually comprised a subpopulation of the culture that failed to reproduce due to starvation-induced injury and hydrogen peroxide sensitivity (Bogosian & Bourneuf, 2001). Interestingly, A. hydrophila incubated in freshwater also entered a VBNC state at 5 u C (Wai et al, 2000;Mary et al, 2002). Controversial results were obtained from attempts to restore culturability via reagents that degrade reactive oxygen species (Wai et al, 2000;Mary et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

Temperature and growth-phase effects on Aeromonas hydrophila survival in natural seawater microcosms: role of protein synthesis and nucleic acid content on viable but temporarily nonculturable response

2004

View full text Add to dashboard Cite

The behaviour of Aeromonas hydrophila in nutrient-poor filter-sterilized seawater was investigated at 23 and 5 6C with respect to its growth phase. At both temperatures, the culturable A. hydrophila population declined below the detection level (0?1 c.f.u. ml "1 ) after 3-5 weeks, depending on the initial physiological state of the cells. During the first week, starved A. hydrophila cells appeared more resistant to the seawater stress at 5 6C than cells initially in the exponential growth phase. This difference was not observed at 23 6C, where de novo protein synthesis seemed to be required for long-term adaptation of cells from the exponential growth phase. Over the duration of the experiments, intact and total cell concentrations were not significantly affected, indicating that bacteria had entered a so-called viable but nonculturable state (VBNC). However, the incubated bacteria rapidly became heterogeneous with respect to their nucleic acid content, and their cell size decreased faster at 23 than at 5 6C. Resuscitation of VBNC cells was attempted by a temperature shift from 5 to 23 6C without exogenous nutrient addition. Comparison of the growth rates of the stressed population and of the untreated bacteria growing in the same autoclaved initial cell suspension showed significantly faster growth for the stressed cells, suggesting that in addition to growth of the few culturable stressed cells, a proportion of injured cells became culturable.

show abstract

Source Water Protection: Microbiology of Source Water

Berger

Oshiro

2003

Encyclopedia of Environmental Microbiology

View full text Add to dashboard Cite

Pathogen Sources Factors Affecting the Transport of Fecal Pathogens to Surface Water Environmental Factors in Pathogen Survival in Ambient Waters Pathogen Occurrence in Surface Waters Groundwater Indicators of Source Water Quality Nonmicrobial Indicators of Fecal Contamination Laws and Regulations Protecting Drinking Water Sources

show abstract

A comparison of solid and liquid media for resuscitation of starvation- and low-temperature-induced nonculturable cells of Aeromonas hydrophila

Cited by 89 publications

References 19 publications

Behavior of an Aeromonas hydrophila aroA Live Vaccine in Water Microcosms

Behavior of an Aeromonas hydrophila aroA Live Vaccine in Water Microcosms

Temperature and growth-phase effects on Aeromonas hydrophila survival in natural seawater microcosms: role of protein synthesis and nucleic acid content on viable but temporarily nonculturable response

Source Water Protection: Microbiology of Source Water

Contact Info

Product

Resources

About