Applied Biosafety

We developed a matrix of changes between Section IV of the current and former editions of Biosafety in Microbiological and Biomedical Laboratories (BMBL), focusing on biosafety levels two, three, and four (BSL2, BSL3, and BSL4). Citations containing multiple statements were subdivided into individually addressable statements and statements with similar/identical scope were aligned, allowing for a precise comparative analysis. In addition, statements were categorized for further analysis based on the subject of the change, the type of change, and the outcome of the change. Although the Fifth Edition of BMBL (BMBL5) proved to be a close facsimile of its predecessor, we identified 375 total changes, including 165 novel statements not addressed in the former edition. Over 65% (110) of these changes were found in statements pertaining to BSL4 containment. In our study, we further examine these edition differences and identify key areas where those entities responsible for compliance with these new recommendations may wish to focus their efforts.

This study evaluated the inactivation of Francisella tularensis Schu S4 on various materials (acrylic, glass, polyamide, polyethylene, polypropylene, silicone rubber, and stainless steel) using hydrogen peroxide fumigation of a Class III Biological Safety Cabinet (BSC III). A suspension of F. tularensis Schu S4 (7 × 10⁷ CFU) was dried on seven different types of test surfaces and exposed to vaporous hydrogen peroxide (VHP) fumigation for a contact time of two hours. Qualitative growth assessment showed that VHP exposure inactivated F. tularensis on all replicates of the seven test materials up to four days post-exposure. The effectiveness of VHP fumigation on the growth of biological indicators (Bacillus subtilis or Geobacillus stearothermophilus) and spore strips (Bacillus atrophaeus) was evaluated in parallel as a qualitative assessment of decontamination. At one and four days post-exposure, decontaminated biological indicators and spore strips exhibited no growth, while the non-decontaminated samples displayed growth. This study provides information for using VHP fumigation as an alternative approach for the decontamination of virulent F. tularensis when the current accepted method of 10% household bleach followed by 70% alcohol may not be practical for decontamination of a BSC III.

Knowledge of the stability of pathogens in the environment is part of a comprehensive biological risk assessment. The inherent nature of laboratory equipment and laboratory procedures to create aerosolized droplets of infectious agents, with subsequent deposition of these particles, provides opportunities to contaminate fomites such as laboratory equipment and personal items. Vivaria create an increased level of virus amplification and fomite contamination as the dried waste of virus-infected research animals can become aerosolized, or transmitted through direct contact. This paper provides a review of studies in which several species of virus have been allowed to dry on porous, or nonporous substrates, and analyzed at timed intervals to determine the ranges over which these viruses remain stable at room temperature as measured by their viability to infect cell cultures or research animals. The research shows that some viruses retain viability for up to a month or longer in the laboratory environment. This review will aid investigators and biosafety professionals in both risk assessment as well as decontamination efforts.

T cell response precedes serological response, and is more feasible than pathogen detection by molecular tools. Thus, monitoring T cell response to class-A pathogens may allow for a timely treatment and a correct handling of patients containment issues, after exposure to potentially lethal agents, such as biological threat, or emerging pathogens. In this paper, we describe a procedure for the evaluation of T cell response to microbial pathogens, including class-A pathogens in BSL-3 and BSL-4 equipped laboratories.

The development and validation of the Biosafety Level 2 (BSL-2) enhanced containment system located at the produce pilot plant facility of the U.S. Department of Agriculture—Eastern Regional Research Center is presented here. This multi-purpose containment is used to enclose pilot and pilot scale washing and sanitizing equipment for fruits and vegetables, or other decontamination equipment where aerosol generation is likely, and complete protection is required for researchers. This containment is operated under a negative pressure with all exhausted air (approximately eight containment air exchanges per hour) being passed in parallel through two hydrophobic HEPA filters. During operation, personnel are excluded from the containment, materials are introduced into and removed from the containment via pass-throughs, and equipment is operated via computer control and glove ports.

At the completion of any series of processing trials, vegetative bacterial cells remaining within the containment, and in the processing water, are inactivated by raising the internal temperature of the containment, and all contents, including equipment, processing water and waste water to sufficiently inactivate all bacterial cells, except spore formers, using industrial steam at atmospheric pressure. We have demonstrated the feasibility and safety of conducting multiple trials using pathogenic bacteria with pilot scale processing equipment within the containment, and then inactivating surviving vegetative bacterial cells with the steam-in-place process. Furthermore, we validated pilot-scale surface pasteurization processing equipment capable of significantly reducing cell densities (in excess of 5 logs) of Salmonella on inoculated cantaloupes, using this BSL-2 enhanced containment system.

The proposed construction of NIH-NIAID biodefense labs provided an opportunity to study real time developments in the public review process for high-level biocontainment facilities and compare them with literature findings on other biocontainment labs and complex science/technology projects. The goals of this study were to examine the types of issues and concerns raised in the review process, evaluate the importance of different factors in the public debate, and understand what features, if any, arose in situations with and without controversies. Based on an analysis of environmental impact documents, detailed timelines and media and Internet communications, it was possible to identify common issues and features associated with the lab reviews. Issues of trust and transparency, concerns about secret, or classified research, undisclosed accidents and a lack of due process were repeatedly found in controversial situations. The lessons learned from this study are relevant to current and future biocontainment projects, and highlight the importance of developing trust, implementing comprehensive early proactive risk communication plans, and maintaining open communication even after operations begin. Current governmental restrictions on transparency and openness associated with biodefense and terrorism make communication increasingly difficult, and have implications for long-term public trust and perceptions about biosecurity.