Use of flow cytometry for analysis of phage-mediated killing of Enterobacter aerogenes

Verthé, Kristof; Verstraete, Willy

doi:10.1016/j.resmic.2006.02.007

Cited by 17 publications

(8 citation statements)

References 16 publications

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“…From a food safety application perspective, the ability of the selected phage to target host cells under refrigerated storage is important (20). The activity of a selected phage at a lower temperature depends on its successful adsorption to the host bacterium's surfaces and active function (20,37). L. monocytogenes cells are able to grow under refrigerated temperature conditions, and are metabolically active at such low temperatures.…”

Section: Discussionmentioning

confidence: 99%

Bacteriophage Significantly Reduces Listeria monocytogenes on Raw Salmon Fillet Tissue

Soni

Nannapaneni

2010

Journal of Food Protection

101

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We have demonstrated the antilisterial activity of generally recognized as safe (GRAS) bacteriophage LISTEX P100 (phage P100) on the surface of raw salmon fillet tissue against Listeria monocytogenes serotypes 1/2a and 4b. In a broth model system, phage P100 completely inhibited L. monocytogenes growth at 4 degrees Celsius for 12 days, at 10 degrees Celsius for 8 days, and at 30 degrees Celsius for 4 days, at all three phage concentrations of 10(4), 10(6), and 10(8) PFU/ml. On raw salmon fillet tissue, a higher phage concentration of 10(8) PFU/g was required to yield 1.8-, 2.5-, and 3.5-log CFU/g reductions of L. monocytogenes from its initial loads of 2, 3, and 4.5 log CFU/g at 4 or 22 degrees Celsius. Over the 10 days of storage at 4 degrees Celsius, L. monocytogenes growth was inhibited by phage P100 on the raw salmon fillet tissue to as low as 0.3 log CFU/g versus normal growth of 2.6 log CFU/g in the absence of phage. Phage P100 remained stable on the raw salmon fillet tissue over a 10-day storage period, with only a marginal loss of 0.6 log PFU/g from an initial phage treatment of 8 log PFU/g. These findings illustrate that the GRAS bacteriophage LISTEX P100 is listericidal on raw salmon fillets and is useful in quantitatively reducing L. monocytogenes.

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Section: Discussionmentioning

confidence: 99%

Bacteriophage Significantly Reduces Listeria monocytogenes on Raw Salmon Fillet Tissue

Soni

Nannapaneni

2010

Journal of Food Protection

101

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“…These applications have been successfully used by many other investigators. Verthé & Verstraete (2006) used PI for the analysis of a bacteriophage‐infected Enterobacter aerogenes strain as an evaluation of phage therapy functioning. Temporary permeabilization by electroporation was also described (Garcia et al ., 2007).…”

Section: Functional Informationmentioning

confidence: 99%

Functional single-cell analyses: flow cytometry and cell sorting of microbial populations and communities

2010

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The still poorly explored world of microbial functioning is about to be uncovered by a combined application of old and new technologies. Bacteria, especially, are still in the dark with respect to their phylogenetic affiliations as well as their metabolic capabilities and functions. However, with the advent of sophisticated flow cytometric and cell sorting technologies in microbiological labs, there is now the possibility to gain this knowledge at the single-cell level without cumbersome cultivation approaches. Cytometry also facilitates the understanding of physiological diversity in seemingly likewise acting populations. Both individuality and diversity lead to the complex and concerted actions of microbial consortia. This review provides an overview of the state of the art in the field. It deals with the handling of microorganisms from the very beginning (i.e. sampling, and detachment and fixation procedures) and goes on to discuss the pitfalls and problems in analysing cells without any further treatment. If information cannot be gained by specific staining procedures, phylogenetic technologies, transcriptomic and proteomic approaches may be options for achieving advanced insights. All in all, flow cytometry will be a mediator technology to gain a deeper insight into the heterogeneity of populations and the functioning of microbial communities.

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“…FDA is usually used to detect viable bacteria (14)(15)(16). The evaluation of microbial viability represents an important requirement in different areas of microbiology, such as health, biotechnologies, food technologies, water industry and pharmaceutical industries (17)(18)(19)(20)(21). The ability to distinguish between different physiological states (viable, latent, non-viable) represents an important tool, especially for the evaluation of survival rates of pathogenic microorganisms (22), of the growth and development of microorganisms in oligotrophic environments (723 and of the effect of toxic substances on microbial activity (24).…”

Section: Introductionmentioning

confidence: 99%

The Influence of Electromagnetic Field on Morphophysiological Parameters of Bacterial Cells Evaluated Through Flow Cytometry

Radu¹,

Marinescu²,

Savin³

et al. 2016

Simi 2016

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Flow cytometry represents an important tool in environmental biology, and especially in cellular biology, through its ability to distinguish among different physiological states (viable, latent and non-viable). Flow cytometry combined with fluorescent markers, such as propidium iodide and ethidium bromide allows to perform rapid measurements on individual cells and also simultaneous measurements of multiple cellular parameters, both structural and functional. The aim of this paper was to investigate the influence of the electromagnetic field on the morpho-physiological parameters of some bacterial strains cells isolated from wastewater, assessed through flow cytometry. In this respect, a number of 10 bacterial strains isolated from wastewater (E. coli, Salmonella sp., Enterobacter sp., Citrobacter sp., Klebsiella sp. and Enterococcus sp.) were exposed to an electromagnetic field (50Hz electric field at different voltages) for 24 hours, at 37°C. Both samples, the ones exposed to electromagnetic field and blank samples (unexposed) were assessed by flow cytometry technique. Two possible mechanism of action have been tracked, i.e. the efflux pumps activity (fluorescence markerEthidium Bromide) and permeabilisation of cellular layers (fluorescence marker -Propidium Iodide). Both mechanisms of action have been identified, from slightly to significant modifications, as compared to untreated controls, quantified as ΔMFI (median of fluorescence intensity). Conclusion: Our results suggest that flow cytometry could be used for the real-time evaluation of the influence of different electromagnetic fields on the aquatic microbiota, the obtained results being of great interest for the development of technological solution with increased efficiency in the (waste)water treatment.

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Use of flow cytometry for analysis of phage-mediated killing of Enterobacter aerogenes

Cited by 17 publications

References 16 publications

Bacteriophage Significantly Reduces Listeria monocytogenes on Raw Salmon Fillet Tissue

Bacteriophage Significantly Reduces Listeria monocytogenes on Raw Salmon Fillet Tissue

Functional single-cell analyses: flow cytometry and cell sorting of microbial populations and communities

The Influence of Electromagnetic Field on Morphophysiological Parameters of Bacterial Cells Evaluated Through Flow Cytometry

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