T1810 Determination of Spatial and Temporal Colonization of Enteropathogenic E. coli and Enterohemorrhagic E. coli in Mice Using Bioluminescent In Vivo Imaging

Rhee, Ki‐Jong; Cheng, Hao; Harris, Antoneicka; Morin, Cara; Kaper, James B.; Hecht, Gail

doi:10.1016/s0016-5085(10)62688-1

Cited by 15 publications

(20 citation statements)

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“…The lux gene derived from bacteria, and luciferase from the firefly and click beetle (luc), are two other common reporter genes used in bioluminescence imaging. Fluorescence imaging is commonly associated with the use of green and red fluorescence proteins [31,32]. Rats and mice are popular model organisms which study the proliferation and colonization of lactic acid bacteria (LAB) [33,34].…”

Section: Fluorescent Reporters and Their Applications For In Vivo Imamentioning

confidence: 99%

“…pTRKH3 is a shuttle vector for E. coli, Enterococcus, Lactococcus, Streptococcus and Lactobacillus [60]. The vector has a copy number (30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40) in E. coli, and a somewhat higher copy number in Lactococcus and Streptococcus species [64]. It carries a gene for erythromycin resistance, which is expressed in E. coli and LAB.…”

Section: Fluorescent Reporters and Their Applications For In Vivo Imamentioning

confidence: 99%

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The Microbiome of Spodoptera littoralis: Development, Control and Adaptation to the Insect Host

Mazumdar¹,

Teh²,

Boland³

2018

Metagenomics for Gut Microbes

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The symbiotic microbial consortium in the gut of Spodoptera littoralis shows dramatic, but reproducible changes in line with the development of the insect from the egg via six larval instars to the pupa. Since the food is kept constant during development, factors from the insect host and certain microbial symbionts are assumed to control the composition of the microbiome. A GFP-tagged Enterococcus mundtii, one of the major players of the consortium, easily integrates into the microbiome and can be monitored in all gut segments at all developmental stages. The reporter organism can be recovered from the gut using a preparative flow cytometry allowing subsequent RNA extraction for transcriptomic analyses. The transcriptomic profile from the fluorescent Enterococcus cells provides information on the adaptation of the reporter organism to the local gut conditions. The concept of using a fluorescent reporter organism that can be recovered at any time from any area of the intestinal tract will allow a holistic analysis of adaptation strategies used by the microbes to adapt to the insect gut. In combination with the analysis of transcript patterns from the gut membranes, a first insight into the molecular interaction between the insect host and the microbiome can be expected.

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Section: Fluorescent Reporters and Their Applications For In Vivo Imamentioning

confidence: 99%

Section: Fluorescent Reporters and Their Applications For In Vivo Imamentioning

confidence: 99%

The Microbiome of Spodoptera littoralis: Development, Control and Adaptation to the Insect Host

Mazumdar¹,

Teh²,

Boland³

2018

Metagenomics for Gut Microbes

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“…The most popular include bioluminescence with bacterial lux genes (lux), firefly and click beetle luciferase (luc), and fluorescence recorded in the case of green and red proteins. 22,31,32 Other well established imaging systems include magnetic resonance imaging (MRI) and positron emission tomography. 25,33 In vivo Fluorescence…”

Section: Reporter Systemsmentioning

confidence: 99%

“…66 The bacterial luciferase system is predominantly used when bacterial infection models are studied. 30,67 An example of this is the cloning of the P. luminescence lux operon into infectious pathogens such as E. coli, Citrobacter rodentium, Listeria monocytogenes and Salmonella spp [20][21][22][23] Pathogenicity of the labeled bacteria remained unaffected and they were visualized throughout infection. Bacteria labeled with the lux genes were detected in the GIT of mice (Fig.…”

Section: In Vivo Bioluminescencementioning

confidence: 99%

“…The same technology was used to study the spread of Listeria monocytogenes infection in a murine model 20 and later by many other research groups to study the migration of bacteria in the GIT. [21][22][23] In this review, the advantages and disadvantages of different in vivo reporter systems are discussed, with emphasis on bioluminescent and fluorescent proteins.…”

Section: Introductionmentioning

confidence: 99%

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Reporter systems forin vivotracking of lactic acid bacteria in animal model studies

2015

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Bioluminescence (BLI) and fluorescence imaging (FI) allow for non-invasive detection of viable microorganisms from within living tissue and are thus ideally suited for in vivo probiotic studies. Highly sensitive optical imaging techniques detect signals from the excitation of fluorescent proteins, or luciferase-catalyzed oxidation reactions. The excellent relation between microbial numbers and photon emission allow for quantification of tagged bacteria in vivo with extreme accuracy. More information is gained over a shorter period compared to traditional pre-clinical animal studies. The review summarizes the latest advances in in vivo bioluminescence and fluorescence imaging and points out the advantages and limitations of different techniques. The practical application of BLI and FI in the tracking of lactic acid bacteria in animal models is addressed.

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Application of Bioluminescence Imaging (BLI) to the Study of the Animal Models of Human Infectious Diseases

Golding

Zaitseva

2013

Pharmaco-Imaging in Drug and Biologics Development

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Traditional approaches in measuring the effi cacy of antimicrobials, antivirals, antifungals, and antiparasitic therapies have limitations which contribute to noise in effi cacy assessment. These include the following: (1) large number of animals must be sacrifi ced at multiple time points to assess the effects of therapy on pathogen titers in the organs; (2) weight loss of 25-30 % in animals requires ethical animal euthanasia which may be unscheduled and skew sampling; and (3) the organ titers are based upon ex vivo culture or analysis. Such limitations can lead to erroneous conclusions regarding drug effi cacy versus spontaneous recovery. One needs to evaluate dissemination of the pathogen in vivo within the same animal to truly measure effi cacy. Bioluminescence imaging (BLI) has been developed as a new method to monitor infections in small animal models. BLI is based on noninvasive measurement of biomarkers possessing a light-producing luciferin or other light-emitting metabolic substrates. This imaging platform is now a widely used technique in oncology, tumor metastasis, establishing effi cacy of anticancer and anti-infective therapies, detecting protein-protein interactions, detecting transgene expression in vivo, and many more. BLI, importantly, enables each animal to be used as its own control over time, thus limiting the number of animals required in studies and minimizing animal-to-animal variations which improves statistical precision of effi cacy outcomes. In this chapter we describe applications of BLI in the study of infection, reveal important limitations of the technique, and summarize recent work in murine microbial and viral models.

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T1810 Determination of Spatial and Temporal Colonization of Enteropathogenic E. coli and Enterohemorrhagic E. coli in Mice Using Bioluminescent In Vivo Imaging

Cited by 15 publications

References 0 publications

The Microbiome of Spodoptera littoralis: Development, Control and Adaptation to the Insect Host

The Microbiome of Spodoptera littoralis: Development, Control and Adaptation to the Insect Host

Reporter systems forin vivotracking of lactic acid bacteria in animal model studies

Application of Bioluminescence Imaging (BLI) to the Study of the Animal Models of Human Infectious Diseases

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