Summary
Whole body biophotonic imaging (BPI) is a technique that has contributed significantly to the way researchers study bacterial pathogens and develop pre-clinical treatments to combat their ensuing infections in vivo. Not only does this approach allow disease profiles and drug efficacy studies to be conducted non-destructively in live animals over the entire course of the disease, but in many cases, it enables investigators to observe disease profiles that could otherwise easily be missed using conventional methodologies. The principles of this technique are that bacterial pathogens engineered to express bioluminescence (visible light) can be readily monitored from outside of the living animal using specialized low-light imaging equipment, enabling their movement, expansion and treatment to be seen completely non-invasively. Moreover, because the same group of animals can be imaged at each time-point throughout the study, the overall number of animals used is dramatically reduced, saving lives, time, and money. Also, as each animal acts as its own control over time, the issues associated with animal-to-animal variation are circumvented, thus improving the quality of the biostatistical data generated. The ability to monitor infections in vivo in a longitudinal fashion is especially appealing to assess chronic infections such as those involving implanted devices. Typically, bacteria grow as biofilms on these foreign bodies and are reputably difficult to monitor with conventional methods. Because of the non-destructive and non-invasive nature of BPI, the procedure can be performed repeatedly in the same animal, allowing the biofilm to be studied in situ without detachment or disturbance. This ability not only allows unique patterns of disease relapse to be seen following termination of antibiotic therapy but also in vivo resistance development during prolonged treatment, both of which are common occurrences with device-related infections. This chapter describes the bioluminescent engineering of both Gram-positive and Gram-negative bacteria and overviews their use in device-associated infections in several anatomical sites in a variety of animal models.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Contag, C. H., Contag, P. R., Mullins, J. I., Spilman, S. D., Stevenson, D. K., and Benaron, D. A.. (1995) Photonic detection of bacterial pathogens in living hosts. Mol. Microbiol. 18, 593–603.
Burns-Guydish, S. M., Olomu, I. N., Zhao, H., Wong, R. J., Stevenson, D. K., and Contag, C. H. (2005) Monitoring age-related susceptibility of young mice to oral Salmonella enterica serovar Typhimurium infection using an in vivo murine model. Pediatr. Res. 58, 153–158.
Francis, K. P., Yu, J., Bellinger-Kawahara, C., Joh, D., Hawkinson, M. J., Xiao, G., Purchio, T. F., Caparon, M. G., Lipsitch, M., and Contag, P. R. (2001) Visualizing pneumococcal infections in the lungs of live mice using bioluminescent Streptococcus pneumoniae transformed with a novel gram-positive lux transposon. Infect. Immun. 69, 3350–3358.
Rocchetta, H. L., Boylan, C. J., Foley, J. W., Iversen, P. W., LeTourneau, D.L., McMillian, C. L., Contag, P. R., Jenkins, D. E., and Parr, T. R., Jr. (2001) Validation of a noninvasive, real-time imaging technology using bioluminescent Escherichia coli in the neutropenic mouse thigh model of infection. Antimicrob. Agents Chemother. 45, 129–137.
Francis, K. P., Joh, D., Bellinger-Kawahara, C., Hawkinson, M. J., Purchio, T. F., and Contag, P. R. (2000) Monitoring bioluminescent Staphylococcus aureus infections in living mice using a novel luxABCDE construct. Infect. Immun. 68, 3594–3600.
Orihuela, C. J., Gao, G., McGee, M., Yu, J., Francis, K. P., and Tuomanen, E. (2003) Organ-specific models of Streptococcus pneumoniae disease. Scand J Infect. Dis. 35, 647–652.
Kadurugamuwa, J. L., Sin, L., Albert, E., Yu, J., Francis, K., DeBoer, M., Rubin, M., Bellinger-Kawahara, C., Parr, T. R., Jr., and Contag, P. R. (2003) Direct continuous method for monitoring biofilm infection in a mouse model. Infect. Immun. 71, 882–890.
Kadurugamuwa, J. L., Sin, L.V., Yu, J., Francis, K. P., Kimura, R., Purchio, T., and Contag, P. R. (2003) Rapid direct method for monitoring antibiotics in a mouse model of bacterial biofilm infection. Antimicrob. Agents Chemother. 47, 3130–3137.
Kadurugamuwa, J. L., Sin, L. V., Yu, J., Francis, K. P., Purchio, T. F., and Contag, P. R. (2004) Noninvasive optical imaging method to evaluate postantibiotic effects on biofilm infection in vivo. Antimicrob. Agents Chemother. 48, 2283–2287.
Kadurugamuwa, J. L., Modi, K., Yu, J., Francis, K. P., Purchio, T., and Contag, P. R. (2005) Noninvasive biophotonic imaging for monitoring of catheter-associated urinary tract infections and therapy in mice. Infect. Immun. 73, 3878–3887.
Kuklin, N. A., Pancari, G.D., Tobery, T.W., Cope, L., Jackson, J., Gill, C., Overbye, K., Francis, K. P., Yu, J., Montgomery, D., Anderson, A. S., McClements, W., and Jansen, K. U. (2003) Real-time monitoring of bacterial infection in vivo: development of bioluminescent staphylococcal foreign-body and deep-thigh-wound mouse infection models. Antimicrob. Agents Chemother. 47, 2740–2748.
Xiong, Y. Q., Willard, J., Kadurugamuwa, J. L., Yu, J., Francis, K. P., and Bayer, A. S. (2005) Real-time in vivo bioluminescent imaging for evaluating the efficacy of antibiotics in a rat Staphylococcus aureus endocarditis model. Antimicrob. Agents Chemother. 49, 380–387.
Yu, J., Wu, J., Francis, K. P., Purchio, T. F., and Kadurugamuwa, J. L. (2005) Monitoring in vivo fitness of rifampicin-resistant Staphylococcus aureus mutants in a mouse biofilm infection model. J. Antimicrob. Chemother. 55, 528–534.
Davies, D. (2003) Understanding biofilm resistance to antibacterial agents. Nat. Rev. Drug Discov. 2, 114–122.
Mermel, L. A., Farr, B. M., Sherertz, R. J., Raad, I. I., O’Grady, N., Harris, J. S., and Craven, D. E. (2001) Guidelines for the management of intravascular catheter-related infections. Clin. Infect. Dis. 32, 1249–1272.
Winson, M. K., Swift, S., Hill, P. J., Sims, C. M., Griesmayr, G., Bycroft, B. W., Williams, P., and Stewart, G. S. (1998) Engineering the luxCDABE genes from Photorhabdus luminescens to provide a bioluminescent reporter for constitutive and promoter probe plasmids and mini-Tn5 constructs. FEMS Microbiol. Lett. 163, 193–202.
Rupp, M. E., Ulphani, J. S., Fey, P. D., Bartscht, K., and Mack, D. (1999) Characterization of the importance of polysaccharide intercellular adhesin/hemagglutinin of Staphylococcus epidermidis in the pathogenesis of biomaterial-based infection in a mouse foreign body infection model. Infect. Immun. 67, 2627–2632.
Warren, J. W. (2001) Catheter-associated urinary tract infections. Int. J. Antimicrob. Agents 17, 299–303.
Kurosaka, Y., Ishida, Y., Yamamura, E., Takase, H., Otani, T., and Kumon, H. (2001) A non-surgical rat model of foreign body-associated urinary tract infection with Pseudomonas aeruginosa. Microbiol. Immunol. 45, 9–15.
Dugdale, D. C. and Ramsey, P. G. (1990) Staphylococcus aureus bacteremia in patients with Hickman catheters. Am. J. Med. 89, 137–141.
Rupp, M. E. and Archer, G. L. (1994) Coagulase-negative staphylococci: pathogens associated with medical progress. Clin. Infect. Dis. 19, 231–243; quiz 244–235.
Herrero, M., de Lorenzo, V., and Timmis, K. N. (1990) Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria. J. Bacteriol. 172, 6557–6567.
de Lorenzo, V., Herrero, M., Jakubzik, U., and Timmis, K. N. (1990) Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria. J. Bacteriol. 172, 6568–6572.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Kadurugamuwa, J.L., Francis, K.P. (2008). Bioluminescent Imaging of Bacterial Biofilm Infections In Vivo. In: DeLeo, F.R., Otto, M. (eds) Bacterial Pathogenesis. Methods in Molecular Biology™, vol 431. Humana Press. https://doi.org/10.1007/978-1-60327-032-8_18
Download citation
DOI: https://doi.org/10.1007/978-1-60327-032-8_18
Publisher Name: Humana Press
Print ISBN: 978-1-58829-740-2
Online ISBN: 978-1-60327-032-8
eBook Packages: Springer Protocols