Validation of an Animal Isolation Imaging Chamber for Use in Animal Biosafety Level-3 Containment.

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Live imaging of animals infected with pathogenic microbes poses a contamination risk to equipment, personnel and other animals. A Caliper animal isolation chamber designed for the IVIS(®) Spectrum imaging system was tested as a containment device for mice infected with microbes assigned to animal biosafety level-3 (ABSL-3). A testing protocol was developed by adapting two published standards to test other equipment in high containment environments. The protocol included quantitative leak-testing of the high efficiency particulate air (HEPA) filters, soap bubble testing of the animal isolation chamber, and pressure decay testing of the complete containment system. HEPA filters were > 99.999% efficient (< 0.001% leakage). When attached to the Spectrum at the normal flow rate of oxygen/anesthetic mix (0.25 L/min), the chamber was positively pressurized at 0.11 inches of water (in H(2)O). No leaks were detected by soap bubble testing at flow rates of 0.25 L/min to 2.0 L/min, generating pressures up to 2.90 in H(2)O. (26-fold increase over normal operating pressure). The complete containment system passed pressure decay testing at 2.0 in H(2)O by sustaining 95% of the initial pressure over a 30 minute period.The Caliper animal isolation chamber provides appropriate isolation for the IVIS(®) Spectrum imaging system. When used as a containment device, it must undergo periodic performance testing, as described here, since it operates under positive pressure. The chamber is an appropriate component of ABSL-3 containment when combined with proper administrative controls and work practices. The testing protocol described here can be used to validate containment devices for other imaging systems or animal species.






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Alderman, T Scott, Richard Frothingham and Gregory D Sempowski (2010). Validation of an Animal Isolation Imaging Chamber for Use in Animal Biosafety Level-3 Containment. Applied biosafety : journal of the American Biological Safety Association, 15(2). pp. 62–66. 10.1177/153567601001500203 Retrieved from

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Scott Alderman

Adjunct Associate in the Department of Medicine

Richard Frothingham

Associate Professor of Medicine

Dr. Frothingham is the principal investigator of a research laboratory which studies Mycobacterium tuberculosis, the cause of tuberculosis, and Mycobacterium avium, a closely related bacterium causing serious infections in AIDS patients. We are pursuing two current projects.

The first project aims to develop vaccines against M. avium and M. tuberculosis. We inject mice with candidate plasmid DNA vaccines which produce bacterial proteins in mouse muscle. We use a variety of DNA adjuvants to modify the immune response. We hope to use DNA vaccination to protect against new infections and to modify the course of existing infections. We also hope to identify correlates of vaccine-induced protective immunity.

The second project uses variations in bacterial DNA sequences to identify species and strains. Dr. Frothingham was part of a team of four Duke scientists who used DNA sequence analysis to identify the cause of Whipple's disease. He also identified used DNA sequence to identify a particular group of M. avium strains which cause disseminated infections in AIDS patients. We recently developed a new tuberculosis typing method using variable numbers of tandem DNA repeats. We are applying this new typing method in national and international collaborations.

Dr. Frothingham does not currently conduct clinical trials.

Special areas of expertise include tuberculosis, mycobacteria, strain differentiation, DNA vaccination, and pyrazinamide.

Key words: tuberculosis, mycobacteria, Mycobacterium tuberculosis, Mycobacterium avium, DNA vaccines, tandem repeat DNA, pyrazinamide, mouse

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