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Volume 26, Number 11—November 2020
Research

Validated Methods for Removing Select Agent Samples from Biosafety Level 3 Laboratories

Alexandria E. Kesterson, John E. Craig, Lara J. Chuvala, and Henry S. HeineComments to Author 
Author affiliation: Author affiliation: University of Florida, Orlando, Florida, USA

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Table 1

Maximum bacterial concentrations of select agents in tissues of infected mice*

Agent (reference) Source of samples, bacterial load
Lung, per g Cell pellet, per mL BAL Blood, per mL Overnight culture, per mL
Bacillus anthracis (5,6) <108 Not tested <104 108
Yersinia pestis (7) <1010 Not tested <106 109
Burkholderia mallei (811) <109 Not tested <104 109
Burkholderia pesudomallei (11,12) <108 105 <105 109
Francisella tularensis (13) 107 Not tested <105 109

*BAL, bronchoalveolar lavage.
†References (7) and (8) use a different strain of B. mallei 
‡Value determined through in-house testing of lung samples.

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References
  1. US Department of Health and Human Services. Biosafety in microbiological and biomedical laboratories, 5th ed. Washington (DC): The Department; 2009.
  2. Centers for Disease Control and Prevention; Animal and Plant Health Inspection Service. Guidance on the Inactivation or Removal of Select Agents or Toxins for Future Use. 2018 [cited 2019 Aug 4]. https://www.selectagents.gov/irg-intro.html
  3. US Government Accountability Office. Actions Needed to Improve Management of DOD's Biosafety and Biosecurity Program. 2018 [cited 2019 Aug 4]. https://www.gao.gov/products/GAO-18-422
  4. Leighton  TJ, Doi  RH. The stability of messenger ribonucleic acid during sporulation in Bacillus subtilis. J Biol Chem. 1971;246:318995.PubMed
  5. Heine  HS, Shadomy  SV, Boyer  AE, Chuvala  L, Riggins  R, Kesterson  A, et al. Evaluation of combination drug therapy for treatment of antibiotic-resistant inhalation anthrax in a murine model. Antimicrob Agents Chemother. 2017;61:e0078817. DOIPubMed
  6. Heine  HS, Bassett  J, Miller  L, Hartings  JM, Ivins  BE, Pitt  ML, et al. Determination of antibiotic efficacy against Bacillus anthracis in a mouse aerosol challenge model. Antimicrob Agents Chemother. 2007;51:13739. DOIPubMed
  7. Heine  HS, Chuvala  L, Riggins  R, Hurteau  G, Cirz  R, Cass  R, et al. Natural history of Yersinia pestis pneumonia in aerosol-challenged BALB/c mice. Antimicrob Agents Chemother. 2013;57:20105. DOIPubMed
  8. Judy  BM, Whitlock  GC, Torres  AG, Estes  DM. Comparison of the in vitro and in vivo susceptibilities of Burkholderia mallei to Ceftazidime and Levofloxacin. BMC Microbiol. 2009;9:88. DOIPubMed
  9. Mott  TM, Johnston  RK, Vijayakumar  S, Estes  DM, Motamedi  M, Sbrana  E, et al. Monitoring therapeutic treatments against Burkholderia infections using imaging techniques. Pathogens. 2013;2:383401. DOIPubMed
  10. Moustafa  DA, Scarff  JM, Garcia  PP, Cassidy  SKB, DiGiandomenico  A, Waag  DM, et al. Recombinant salmonella expressing Burkholderia mallei LPS O antigen provides protection in a murine model of melioidosis and glanders. PLoS One. 2015;10:e0132032. DOIPubMed
  11. Lafontaine  ER, Zimmerman  SM, Shaffer  TL, Michel  F, Gao  X, Hogan  RJ. Use of a safe, reproducible, and rapid aerosol delivery method to study infection by Burkholderia pseudomallei and Burkholderia mallei in mice. PLoS One. 2013;8:e76804. DOIPubMed
  12. Tan  GG, Liu  Y, Sivalingam  SP, Sim  S-H, Wang  D, Paucod  J-C, et al. Burkholderia pseudomallei aerosol infection results in differential inflammatory responses in BALB/c and C57Bl/6 mice. J Med Microbiol. 2008;57:50815. DOIPubMed
  13. Heine  HS, Chuvala  L, Riggins  R, Cirz  R, Cass  R, Louie  A, et al. Natural history of Francisella tularensis in aerosol-challenged BALB/c mice. Antimicrob Agents Chemother. 2016;60:183440. DOIPubMed
  14. Dauphin  LA, Bowen  MD. A simple method for the rapid removal of Bacillus anthracis spores from DNA preparations. J Microbiol Methods. 2009;76:2124. DOIPubMed

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