Viable but non‐culturable salmonellas in soil

Turpin, Phillip E.; Maycroft, Kevin A.; Rowlands, Christopher; Wellington, Elizabeth M. H.

doi:10.1111/j.1365-2672.1993.tb05149.x

Cited by 82 publications

(62 citation statements)

References 38 publications

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“…Similar observations have been made with other soil microbes including Flavobacterium sp. strain P25 (13), P. fluorescens (33), Salmonella (29), and X. campestris pv. campestris (10).…”

Section: Discussionmentioning

confidence: 99%

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The Viable But Nonculturable State of Ralstonia solanacearum May Be Involved in Long-Term Survival and Plant Infection

Grey

Steck

2001

Appl Environ Microbiol

167

120

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The role of the dormant-like viable but nonculturable (VBNC) condition in the etiology of bacterial infection was examined using a plant system. The plant-pathogenic bacterium Ralstonia solanacearum was first shown to enter into the VBNC state both in response to cupric sulfate when in a saline solution and when placed in autoclaved soil. To determine if the VBNC condition is related to pathogenesis, the physiological status of bacteria recovered from different regions of inoculated tomato plants was determined at different stages of infection. The fraction of in planta bacteria that were VBNC increased during infection and became greater than 99% by the late stage of disease. The possibility that soil-dwelling VBNC bacteria may resuscitate and infect plants was also examined. When tomato seeds were germinated in sterile soil that contained VBNC but no detectable culturable forms of R. solanacearum cells, resuscitation was observed to occur in soil adjacent to plant roots; these resuscitated bacteria were able to infect plants. This is the first report of R. solanacearum entering the VBNC state and of resuscitation of any VBNC plant-pathogenic bacteria and provides evidence that the VBNC state may be involved in explaining the persistent nature of some infections.

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“…Similar observations have been made with other soil microbes including Flavobacterium sp. strain P25 (13), P. fluorescens (33), Salmonella (29), and X. campestris pv. campestris (10).…”

Section: Discussionmentioning

confidence: 99%

“…vesicatoria, and X. campestris pv. campestris (2,4,10,20,28,29,30,33). That soil microbes can become VBNC in soil may at least partly explain the observation that the percentage of cells present in soil samples that can be recovered in a culturable form is usually very low (0.01 to 10%) (5,24).…”

mentioning

confidence: 99%

The Viable But Nonculturable State of Ralstonia solanacearum May Be Involved in Long-Term Survival and Plant Infection

Grey

Steck

2001

Appl Environ Microbiol

167

120

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“…Its application to terrestrial environments such as soil and compost, however, is very limited. The DVC method reportedly did not work well on Salmonella in soil samples (29). Clay particles and/or organic matter in soil and compost may partly account for the failure.…”

Section: Discussionmentioning

confidence: 99%

“…The VNC pathogens have received a great deal of attention as their existence may pose a potential risk to human health (23). As for agricultural environments such as soil and manure, there is still no evidence that fecal pathogens enter into a VNC state, although it is quite possible (29), mostly due to the lack of appropriate culture-independent techniques to detect viable cells of specific bacteria in such complicated environments which contain large amounts of clay minerals or organic matter that interfere with detection.…”

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confidence: 99%

Direct Viable Count Combined with Fluorescence in situ Hybridization (DVC-FISH) for Specific Enumeration of Viable Escherichia coli in Cow Manure

Ueno

Inoue

et al. 2009

Microb. Environ.

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A direct viable count procedure combined with fluorescence in situ hybridization (DVC-FISH) was developed for the specific detection and enumeration of viable Escherichia coli in cow manure. The DVC method was performed by trapping bacterial cells, extracted from cow manure samples, onto Nucleopore filters followed by incubation on a DVC medium containing yeast extract and four gyrase inhibitors. E. coli cells were identified by using the probe ES445. The DVC method efficiently promoted the elongation of E. coli cells and allowed for the recognition of individual cell division events, by observing microcolonies. Cell recovery by DVC-FISH together with bacterial extraction, was 53% with an inoculum of 10 7 to 10 10 cells g −1 dry weight, when the manure samples were inoculated with a fresh culture of E. coli and determinations were made immediately. An examination of the survival of E. coli in a cow manure microcosm showed that an increasing fraction of E. coli became non-culturable but were still detectable by DVC-FISH. All these results suggest that DVC-FISH is useful for enumerating viable, even non-culturable, E. coli in cow manure.

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“…A high cell/bead ratio is suitable for studies with soil due to binding of cells to soil particles and reduced mixing. In this study, nonspecific binding of antibody to soil particles was reduced by use of gelatin as a blocking agent; this technique has been used for immunofluorescence studies of soil for Rbixobium (Bohool & Schmidt, 1980;Postama e t al., 1988 ;Schmidt, 1974) and Salmonella (Turpin et al , 1993a).…”

Section: Discussionmentioning

confidence: 99%

Selective recovery of Streptosporangium fragile from soil by indirect immunomagnetic capture

Mullins

Gürtler²,

Wellington³

1995

Microbiology

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A polyclonal anti body raised to Stmptosporangium fkagi/e spores reacted strongly and specifically with the immunizing strain and to a number of related species of Streptosporangium, as determined by dot immunoblotting. An indirect immunomagnetic capture method was developed for the recovery of the target organism from sterile and non-sterile soil, using sheep anti-rabbit M-280 Dynabeads. The effects of different soil blocking agents, antibody labelling concentrations and spore/Dynabead capture times on the recovery of 5. fragile spores were investigated. Pre-blocking of antibody binding sites within the soil, with either 2% partially hydrolysed gelatin or 10% skimmed milk, was essential prior to immunomagnetic capture. Increasing the capture time from 15 to 60 min did not affect spore recovery; however, a 10-fold decline in the magnetic bead concentration did result in a significantly lower recovery of spores from soil. 5. fragi/e was selectively enriched (1 :190=fold) when present as a mixed population with Arthmbacter oxydans in sterile soil. The indirect immunomagnetic capture method was used to selectively recover 5. fragile spores seeded into non-sterile soil, although some background binding of non-target bacteria was noted. The target was successfully recovered from a sterile soil microcosm after 14 d incubation and the capture rate was increased by the inclusion of an initial soil dispersion and biomass concentration procedure, using the ion-exchange resin Chelex 100.Keywords : immunocapture, Streptosporangium, soil, magnetic beads, selective isolation INTRODUCTIONThere is an increasing need to improve methods for the detection and isolation of bacteria from environmental samples, both for the recovery of genetically engineered strains and microbial inoculants (Ford & Olsen, 1988;Morgan et al., 1989; Trevors e t al., 1993) and for the isolation of rare and unusual bacteria for use in industrial screening programmes (Nolan & Cross, 1988 ;Shearer, 1987).Immunomagnetic capture (IMC) represents a potentially important development within this field, allowing the specific recovery of target bacteria from a diverse range of environmental samples. The method relies upon the interaction between cell-surface antigens and antibodies which can be attached to magnetized polystyrene beads.Abbreviations: DIB, dot irnrnunoblotting; IMC, irnrnunornagnetic capture; MPC, magnetic partlcle concentrator; Non, Nonidet P-40.Separation of target cells from a suspension can be achieved using a magnetic field. IMC has been widely used in clinical studies for the separation or depletion of target cells from a mixed population (Gaudernack et ai., 1986; Gruhn e t al., 1991 ; Silvestri e t al., 1992). More recently it has been used to enrich for and enhance the detection of pathogenic bacteria present in contaminated foodstuffs (Johne e t al., 1989; Mansfield e t a!., 1993; Parmer eta/., 1992; Skjerve etai., 1990) and faeces (Luk & Lindberg, 1991 ; Tomoyasu, 1992). Work has also focused on the detection and isolation of bacteri...

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Viable but non‐culturable salmonellas in soil

Cited by 82 publications

References 38 publications

The Viable But Nonculturable State of Ralstonia solanacearum May Be Involved in Long-Term Survival and Plant Infection

The Viable But Nonculturable State of Ralstonia solanacearum May Be Involved in Long-Term Survival and Plant Infection

Direct Viable Count Combined with Fluorescence in situ Hybridization (DVC-FISH) for Specific Enumeration of Viable Escherichia coli in Cow Manure

Selective recovery of Streptosporangium fragile from soil by indirect immunomagnetic capture

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