Viability PCR, a Culture-Independent Method for Rapid and Selective Quantification of Viable
            <i>Legionella pneumophila</i>
            Cells in Environmental Water Samples

Delgado‐Viscogliosi, Pilar; Solignac, Lydie; Delattre, J. M.

doi:10.1128/aem.02878-08

Cited by 134 publications

(103 citation statements)

References 60 publications

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“…The event of L. pneumophila being viable but non-culturable (VBNC) is fairly common during water treatment processes and has been documented after chlorination or other high levels of oxidative or physical stress, such as ultraviolet radiation and heat, by multiple studies [47,[125][126][127]. It has been theorized that the bacteria become VBNC either because the cell membrane is not permeable at low chlorine levels, and the cells are stressed or injured during treatment rather than inactivated.…”

Section: Effects Of Engineered System Stressors On L Pneumophilamentioning

confidence: 99%

“…L. pneumophila is tolerant of low-nutrient environments but does require iron to survive and thrive [45,46]. In harsh environments, the bacteria can enter a viable but non-culturable (VBNC), state in which it will not replicate, but can be resuscitated in more friendly conditions [47].…”

Section: Microbiology Of Legionella Sppmentioning

confidence: 99%

“…It has been theorized that the bacteria become VBNC either because the cell membrane is not permeable at low chlorine levels, and the cells are stressed or injured during treatment rather than inactivated. Or the membrane is permeabilized early, and the hole allows ethidium monoazide bromide to enter the cell and prevent reproduction, but the polymerase chain reaction (PCR) signal is not always reduced [47]. Both heat and chlorination can result in damage to the cell membrane, causing the cell to become VBNC.…”

Section: Effects Of Engineered System Stressors On L Pneumophilamentioning

confidence: 99%

“…The term "free chlorine" refers to HOCl and OCl − , though HOCl is known to be more biocidal. Free chlorine inactivates bacteria by adversely affecting their respiratory and transport activities, causing deleterious effects on bacterial membranes, and causing direct oxidative damage to proteins and nucleotide bases [47,120]. Free-living L. pneumophila were killed within 3 min of being exposed to an aqueous solution of 2 mg/L of chlorine, whereas L. pneumophila living within biofilms were much more difficult to treat utilizing chlorine [121].…”

Section: Effects Of Engineered System Stressors On L Pneumophilamentioning

confidence: 99%

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Knowledge to Predict Pathogens: Legionella pneumophila Lifecycle Critical Review Part I Uptake into Host Cells

Mraz

Weir

2018

Water

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Legionella pneumophila (L. pneumophila) is an infectious disease agent of increasing concern due to its ability to cause Legionnaires' Disease, a severe community pneumonia, and the difficulty in controlling it within water systems. L. pneumophila thrives within the biofilm of premise plumbing systems, utilizing protozoan hosts for protection from disinfectants and other environmental stressors. While there is a great deal of information regarding how L. pneumophila interacts with protozoa and human macrophages (host for human infection), the ability to use this data in a model to attempt to predict a concentration of L. pneumophila in a water system is not known. The lifecycle of L. pneumophila within host cells involves three processes: uptake, growth, and egression from the host cell. The complexity of these three processes would risk conflation of the concepts; therefore, this review details the available information regarding how L. pneumophila invades host cells (uptake) within the context of data needed to model this process, while a second review will focus on growth and egression. The overall intent of both reviews is to detail how the steps in L. pneumophila's lifecycle in drinking water systems affect human infectivity, as opposed to detailing just its growth and persistence in drinking water systems.

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Section: Effects Of Engineered System Stressors On L Pneumophilamentioning

confidence: 99%

Section: Microbiology Of Legionella Sppmentioning

confidence: 99%

Section: Effects Of Engineered System Stressors On L Pneumophilamentioning

confidence: 99%

Section: Effects Of Engineered System Stressors On L Pneumophilamentioning

confidence: 99%

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Knowledge to Predict Pathogens: Legionella pneumophila Lifecycle Critical Review Part I Uptake into Host Cells

Mraz

Weir

2018

Water

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“…However, it is complex and time consuming, need 3 to 10 days. Although there has been some rapid and convenient means such as polymerase chain reaction (PCR) and quantitative polymerase chain reaction (qPCR) to detect Legionella in water samples, there powerless to distinguish live and dead bacteria cells limits their application (Delgado et al, 2009). Therefore, culture method still shows its vitality.…”

Section: Introductionmentioning

confidence: 99%

Comparative study on sampling methods for monitoring Legionella species in environmental water

Zhan¹,

Hu²,

Zhu³

2014

Afr. J. Microbiol. Res.

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Infections caused by Legionella spp. are considered an emerging public health problem and are linked to high rates of mortality and morbidity, if not properly treated. This study analyzes 76 samples of water in Guangzhou with the aim of obtaining a more efficient means based on culture to isolate the genus Legionella. A comparative assessment of techniques recommended was performed on monitoring Legionella spp. in environmental water samples. Fourteen (14) sampling methods included in four categories based on concentration (filtration or centrifugation), acid buffer treatment (no acid treatment, treatment for 10 min), thermal treatment (no thermal treatment or treated for 30 min in 50°C) and heat enrich the samples in 37°C for 72 h were tested to identify their sensitivity. The four categories methodologies were assembled by different procedures and 14 methods were obtained to compare the efficiency for isolating Legionella in 76 water samples and in which Legionella were isolated from 42 water samples through these 14 methods. Merit and demerit of each single method was evaluated. Results suggest that a single category methodology for water sampling was insufficient to isolate Legionella spp. from water samples. Some Legionella positive water samples were missed through a single category methodology which made the results mistranslation. PSF category methods in this study were recommended methods and they could isolate Legionella species from 78.6% Legionella positive water samples; and if time was enough, PSF category methods combined with ERCT category methods to isolate Legionella from water samples are strongly recommended. Legionella were isolated from 41 water samples through these two category methods, it only missed 2.4% positive water sample. The results obtained, highlighted the importance of improving culture method for the identification of Legionella in water samples.

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References

2010

Wastewater Microbiology

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Viability PCR, a Culture-Independent Method for Rapid and Selective Quantification of Viable Legionella pneumophila Cells in Environmental Water Samples

Abstract: PCR-based methods have been developed to rapidly screen for

Cited by 134 publications

References 60 publications

Knowledge to Predict Pathogens: Legionella pneumophila Lifecycle Critical Review Part I Uptake into Host Cells

Knowledge to Predict Pathogens: Legionella pneumophila Lifecycle Critical Review Part I Uptake into Host Cells

Comparative study on sampling methods for monitoring Legionella species in environmental water

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