Use of Resazurin To Rapidly Enumerate
            <i>Bdellovibrio</i>
            and Like Organisms and Evaluate Their Activities

Jang, Hyochan; Mun, Wonsik; Choi, Seong Yeol; Mitchell, Robert J.

doi:10.1128/spectrum.00825-22

Cited by 8 publications

(3 citation statements)

References 18 publications

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“…enumeration in under one hour have been reported in 2022. The resazurin assay relies on the detection of NADH produced by B. bacteriovorus , which transforms weakly fluorescent resazurin into the highly fluorescent resorufin ( Jang et al, 2022 ). Alternatively, the SYBR Green assay quantifies total DNA content to determine predator concentration ( Remy et al, 2022 ).…”

Section: Latest Methods and Technology On Imaging And Quantificationmentioning

confidence: 99%

Advances in cellular and molecular predatory biology of Bdellovibrio bacteriovorus six decades after discovery

2023

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Since its discovery six decades ago, the predatory bacterium Bdellovibrio bacteriovorus has sparked recent interest as a potential remedy to the antibiotic resistance crisis. Here we give a comprehensive historical overview from discovery to progressive developments in microscopy and molecular mechanisms. Research on B. bacteriovorus has moved from curiosity to a new model organism, revealing over time more details on its physiology and fascinating predatory life cycle with the help of a variety of methods. Based on recent findings in cryo-electron tomography, we recapitulate on the intricate molecular details known in the predatory life cycle including how this predator searches for its prey bacterium, to how it attaches, grows, and divides all from within the prey cell. Finally, the newly developed B. bacteriovorus progeny leave the prey cell remnants in the exit phase. While we end with some unanswered questions remaining in the field, new imaging technologies and quantitative, systematic advances will likely help to unravel them in the next decades.

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Section: Latest Methods and Technology On Imaging And Quantificationmentioning

confidence: 99%

Advances in cellular and molecular predatory biology of Bdellovibrio bacteriovorus six decades after discovery

2023

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“…In a survey looking for bacterial predatory strains within pristine polar regions, strain PAP01 T was isolated from biocrust glacier foreland of Midtre Lovénbreen, Svalbard, Norway, based on protocols described previously [22,23]. Briefly, 3 g biocrust was mixed with 10 ml sterile HEPES buffer (25 mM, pH 7.2 with 3 mM MgCl 2 and 2 mM CaCl 2 ) at 20 °C and 250 r.p.m.…”

Section: Isolation and Ecologymentioning

confidence: 99%

Bdellovibrio svalbardensis sp. nov., a newly described predator isolated from Svalbard, Norway

Choi,

Mun,

Choi

et al. 2024

International Journal of Systematic and Evolutionary Microbiology

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Identified as a newly described species from a biocrust in Svalbard, Norway (78° 54′ 8.27″ N 12° 01′ 20.34″ E), isolate PAP01T has different characteristics from any known predatory bacteria. The isolate was vibrio-shaped strain that employed flagellar motility. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolate clustered within the genus Bdellovibrio in the family Bdellovibrionaceae . 16S rRNA gene sequence similarities between strain PAP01T and the type strain ( Bdellovibrio bacteriovorus HD100) was 95.7 %. The PAP01T genome has a size of 3.898 Mbp and possesses 3732 genes and a G+C content of 45.7 mol%. The results of genetic and physiological tests indicated the phenotypic differentiation of strain PAP01T from the two other Bdellovibrio species with validly published names. Based on the physiological and phylogenetic data, as well as the prey range spectrum and osmolality sensitivities, isolate PAP01T represents a novel species within the genus Bdellovibrio , for which the name Bdellovibrio svalbardensis sp. nov. is proposed. The type strain is PAP01T (=KCTC 92583T=DSM 115080T).

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“…With the progress of new genetic tools and techniques, luminescent and fluorescent prey are applied to indicate changes in the predator viability and prey population [6][7][8][9] . Other methods such as real-time quantitative PCR 10,11 , flow cytometry 12 and fluorescence visualization 13,14 are utilized to detect the presence and viability of the predator. Even these rapid methods are described, PFU (plaques forming unit) counting on the doublelayer agar plate is still the conventional, economic, and standard methods for detect and quantify the predator viability and biomass 13,15,16 .…”

Section: Introductionmentioning

confidence: 99%

An efficient method for visualizing the plaques ofBdellovibrio bacteriovorus

Zhao,

Xu,

Zhu

2024

Preprint

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Bdellovibrio bacteriovoruspreys upon other Gram-negative bacteria. This obligate predator is recognized as a living antibiotic to control the rising problem of antibiotic resistance. Plaque forming units (PFU) counting is commonly used to determine the viable numbers ofB. bacteriovorus. However, nearly 3-days incubation is always necessary for getting the single, obvious plaques on the double layer agar plate. This time-consuming procedure greatly impedes the purification and enumeration efficiency ofB. bacteriovorus. In this work, we evaluated the advantages of fluorescent prey on the plaque visualization of the predator. Our study reveals that plaques of theB. bacteriovorushave already formed before they could be monitored by eyes on the double layer agar plate. The regular single plaques are easily observed under the enhanced background of fluorescent prey lawn in 1.5 days, reducing nearly half of the time consumption in the purification and enumeration ofB. bacteriovorus. In addition, it also provides some paradigms for modeling and testing the subtle predation process on the solid medium.IMPORTANCEThe viability of predatorBdellovibrio bacteriovorusis typically suggested by the lysis of the prey bacteria on the double layer agar plate. However, long period incubation is necessary to get single obvious plaques ofB. bacteriovorus. Here, we found that plaques are already generated before it can be monitored by eyes. The plaques are obvious on the enhanced background of the double layer agar plate in the presence of fluorescent prey under UV light. We confirmed that the utilization of fluorescence labeled prey bacteria reduces the time spent on the isolation, purification, and enumeration of the predatorB. bacteriovoruson the double layer agar plate.

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Use of Resazurin To Rapidly Enumerate Bdellovibrio and Like Organisms and Evaluate Their Activities

Cited by 8 publications

References 18 publications

Advances in cellular and molecular predatory biology of Bdellovibrio bacteriovorus six decades after discovery

Advances in cellular and molecular predatory biology of Bdellovibrio bacteriovorus six decades after discovery

Bdellovibrio svalbardensis sp. nov., a newly described predator isolated from Svalbard, Norway

An efficient method for visualizing the plaques ofBdellovibrio bacteriovorus

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