Establishing an <i>In Vivo</i> Imaging Capability in High-Containment

Flick-Smith, Helen C.; Commander, Nicola J.; Scott, Andrew E.; Thomas, Richard J.; Brown, Mark A.; Richards, Mark I.; Scott, Joanne C.; Martin, Kevin R.; Harding, Sarah V.; Atkins, Helen S.

doi:10.1177/1535676016631878

Cited by 3 publications

(3 citation statements)

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“…For bioimaging, we took advantage of a luciferase-expressing strain of plague, Y. pestis CO92 pGEN-luxCDABE 22 . To determine if the predatory bacteria B. bacteriovorus HD100 were able to protect mice from a lethal challenge of Y. pestis , SKH-1 were pre-treated with B. bacteriovorus HD100:mCherry via the i.p.…”

Section: Resultsmentioning

confidence: 99%

“…Y. pestis CO92 pGEN-luxCDABE 22 was grown for ~16 h in BAB broth supplemented with 50 µg/ml ampicillin at 28 °C, 180 rpm in a shaking incubator in an biosafety level 3 laboratory. This culture was diluted 1 in 10 in fresh BAB broth supplemented with 50 µg/ml ampicillin and grown at 28 °C, 180 rpm in a shaking incubator until OD 590 was 0.6 (an OD which is approximately 1 × 10 8 cfu/ml).…”

Section: Methodsmentioning

confidence: 99%

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Predatory bacteria can protect SKH-1 mice from a lethal plague challenge

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Flick-Smith

Keyser

et al. 2019

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With the rise of antimicrobial resistance, novel ways to treat bacterial infections are required and the use of predatory bacteria may be one such approach. Bdellovibrio species have been shown in vitro to predate on a wide range of other Gram-negative bacteria, including CDC category A/B pathogens such as Yersinia pestis . The data reported here show that treatment of SKH-1 mice with Bdellovibrio bacteriovorus HD100 provided significant protection from a lethal challenge of Yersinia pestis CO92. This is the first report of protection conferred by predation in vivo against a systemic pathogen challenge. However, this protective effect was not observed in a preliminary study with Balb/c mice. Therefore the effects of the predatory bacteria are complex and may be dependent on immune status/genetics of the host. Overall, predatory bacteria may have utility as a therapeutic modality but further work is required to understand the predator-host interaction.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Predatory bacteria can protect SKH-1 mice from a lethal plague challenge

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Flick-Smith

Keyser

et al. 2019

Sci Rep

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“…Following infection with F. tularensis, animals were handled under ACDP (Advisory Committee for Dangerous Pathogens) containment biosafety level III conditions for imaging studies (25). Mice were weighed daily and were observed twice daily for clinical signs of disease.…”

Section: Methodsmentioning

confidence: 99%

A Bioluminescent Francisella tularensis SCHU S4 Strain Enables Noninvasive Tracking of Bacterial Dissemination and the Evaluation of Antibiotics in an Inhalational Mouse Model of Tularemia

Hall

Flick-Smith

Harding

et al. 2016

Antimicrob Agents Chemother

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bBioluminescence imaging (BLI) enables real-time, noninvasive tracking of infection in vivo and longitudinal infection studies. In this study, a bioluminescent Francisella tularensis strain, SCHU S4-lux, was used to develop an inhalational infection model in BALB/c mice. Mice were infected intranasally, and the progression of infection was monitored in real time using BLI. A bioluminescent signal was detectable from 3 days postinfection (3 dpi), initially in the spleen and then in the liver and lymph nodes, before finally becoming systemic. The level of bioluminescent signal correlated with bacterial numbers in vivo, enabling noninvasive quantification of bacterial burdens in tissues. Treatment with levofloxacin (commencing at 4 dpi) significantly reduced the BLI signal. Furthermore, BLI was able to distinguish noninvasively between different levofloxacin treatment regimens and to identify sites of relapse following treatment cessation. These data demonstrate that BLI and SCHU S4-lux are suitable for the study of F. tularensis pathogenesis and the evaluation of therapeutics for tularemia. Francisella tularensis is a Gram-negative intracellular bacterium and is the etiological agent of the disease tularemia. Tularemia has a low infectious dose by the aerosol route (50% infectious dose [ID 50 ], Ͻ10 CFU), causing a disabling illness without rapid treatment, and is considered a potential biothreat agent. The preferred treatments for tularemia are limited to a few antibiotics, including fluoroquinolones (1, 2), tetracycline (3), streptomycin (4, 5), and gentamicin (6, 7). The lack of a licensed vaccine, reported treatment failures, even with preferred antibiotics (8), and a 2% mortality rate despite treatment (5) necessitate the development and evaluation of new therapies.Conventional methods of assessing bacterial growth and dissemination in vivo require the infection of large cohorts of animals and the culling of groups of animals at set time points throughout a study. This strategy not only uses large numbers of animals but is also labor-intensive and subject to tissue sampling bias. Noninvasive imaging techniques, such as bioluminescence imaging (BLI), can report spatial and temporal aspects of infectious diseases in vivo both longitudinally and in real time (9). BLI typically relies on the detection of light produced by luciferase-catalyzed oxidation reactions, which are encoded in the pathogen of interest, by use of a sensitive charge-coupled device (CCD) camera (10). Pivotal experiments by Contag et al. have demonstrated the usefulness of BLI in the noninvasive study of bacterial pathogenesis and the evaluation of antibiotic treatments for Salmonella enterica serovar Typhimurium (11). Subsequently, bioluminescent reporters have been introduced into other bacteria, including Mycobacterium tuberculosis (10, 12), Staphylococcus aureus (13, 14), Burkholderia mallei (15), and Yersinia pestis (16,17). Further, BLI has been used to evaluate vaccines (18), antibiotics (19-21), and supportive therapies (22) for a ran...

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Replacement, Refinement, and Reduction in Animal Studies With Biohazardous Agents

et al. 2018

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Animal models are critical to the advancement of our knowledge of infectious disease pathogenesis, diagnostics, therapeutics, and prevention strategies. The use of animal models requires thoughtful consideration for their well-being, as infections can significantly impact the general health of an animal and impair their welfare. Application of the 3Rs—replacement, refinement, and reduction—to animal models using biohazardous agents can improve the scientific merit and animal welfare. Replacement of animal models can use in vitro techniques such as cell culture systems, mathematical models, and engineered tissues or invertebrate animal hosts such as amoeba, worms, fruit flies, and cockroaches. Refinements can use a variety of techniques to more closely monitor the course of disease. These include the use of biomarkers, body temperature, behavioral observations, and clinical scoring systems. Reduction is possible using advanced technologies such as in vivo telemetry and imaging, allowing longitudinal assessment of animals during the course of disease. While there is no single method to universally replace, refine, or reduce animal models, the alternatives and techniques discussed are broadly applicable and they should be considered when infectious disease animal models are developed.

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Establishing an In Vivo Imaging Capability in High-Containment

Cited by 3 publications

References 8 publications

Predatory bacteria can protect SKH-1 mice from a lethal plague challenge

Predatory bacteria can protect SKH-1 mice from a lethal plague challenge

A Bioluminescent Francisella tularensis SCHU S4 Strain Enables Noninvasive Tracking of Bacterial Dissemination and the Evaluation of Antibiotics in an Inhalational Mouse Model of Tularemia

Replacement, Refinement, and Reduction in Animal Studies With Biohazardous Agents

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