Applied Biosafety

Whole body imaging technologies, coupled with the generation of stable light emitting strains of infectious agents, make it possible to monitor the location and kinetics of agent growth inside an animal by noninvasive nonterminal methodologies throughout the course of the infection. However, imaging of viable hazardous pathogens, such as Yersinia pestis, requires specialized containment facilities to prevent contamination of both personnel and the wider environment. Here we describe the successful establishment of an in vivo bioimaging capability specifically addressing the requirement for safe and secure work with hazard group 3 pathogens and demonstrate its utility for imaging mice infected with a bioluminescence producing strain of Y. pestis.

It is a well-known fact that personnel activities can have an adverse effect on the protective functions of biological safety cabinets. Increased air turbulence within the work area or within the surrounding room can disrupt the essential directional airflows of the cabinet, which generate the protective barrier between the environment and the products handled inside. This may result in an enhanced escape and/or invasion of airborne particles or microorganisms. Nevertheless, manufacturers tend to reduce downflow and inflow velocities up to the limits of the allowed values to improve energy efficiency or to reduce noise and vibration. Reliable data based on standardized test procedures to estimate the consequences arising from these measures are rare. In this study, the influence of different static and dynamic disturbing factors on the personnel protection performance of 2 Class II biological safety cabinets was quantified. A microbiological test procedure given by the relevant European and US standards (EN 12649, NSF/ANSI 49) was used, but in contrast to the low requirements defined there, 4 more complex and realistic test scenarios were chosen to simulate working activities: static covering of the front sash opening caused by a sitting or standing person was simulated by a “dummy worker” and a “body plate,” respectively. Dynamic airflow perturbations were generated by an artificial “moving arm” swinging regularly inside the cabinet and by a flat plate running outside the cabinet parallel to its front opening to simulate a “walking man”. It could be demonstrated that dynamic airflow disturbances caused by rapid body movements have a major impact on the cabinet’s protecting performance. Compared with an undisturbed working situation, personnel-protecting capabilities of both safety cabinets tested declined substantially when the worker’s movement next to the front opening was simulated. Therefore, downflow and inflow velocities should not be reduced to minimum values, to allow a sufficient margin of safety for actual in-use laboratory conditions (“disturbed”).

Micro-Chem Plus (MCP) detergent disinfectant cleaner is a common chemical disinfectant currently used at most biosafety level 4 laboratories to ensure proper disinfection and decontamination of waste contaminated with risk group 4 agents. However, similar to a number of disinfectants, MCP contains surfactants known as nonylphenol ethoxylates that are considered highly toxic to the aquatic environment. An alternate disinfectant, Earth Sense (ES) neutral disinfection detergent, was identified as one that lacks nonylphenol ethoxylates. This study compares the virucidal efficacy profiles of both MCP and ES using Vesicular Stomatitis Virus (VSV) as a surrogate for risk group 4 agents. Results indicate that a 5% solution of ES with a contact time of 5 minutes was comparable to 5% MCP solution and reduced VSV titer by at least 6 log₁₀. In addition, both 5% solutions of ES and MCP disinfectants were stable for at least 5 months and were able to inactivate 6 log₁₀ of VSV. These results demonstrate that ES detergent may be a potential option to replace MCP as a suitable disinfectant against risk group 4 agents due to its comparable virucidal efficacy as well as its greener environmental profile.

Biosafety and biorisk management are emerging areas of safety that are not well understood by the general public. With recent headlines questioning the safety practices and procedures at federal agencies, educating the public on what steps are taken to encourage a safe and transparent working environment in high-containment laboratories is important. This article describes for the ABSA International biosafety community, as well as for the general public, the biorisk program at the National Institutes of Health and discusses the steps taken to train researchers, report injuries, and encourage a culture of safety within the laboratories, rather than a culture of fear. It attempts to dispel the provocative headlines used by the media, through education on the process and procedures used to protect the researchers, as well as the public, when handling agents whose manipulation and propagation are restricted to high-containment laboratories. In doing so, the authors seek to better educate the public and help to create better awareness of the work that occurs in these facilities.

The Tufts New England Regional Biosafety Laboratory facility has been using chlorine dioxide gas for >5 years, and >100 decontaminations have been done at the room level and on a Class III cabinet. The rooms have ranged from animal holding rooms containing animal racks (housing rodents and rabbits), biosafety cabinets, and computers to biosafety level 3 laboratories containing a variety of equipment (microscopes, biosafety cabinets, centrifuges, incubators, real-time polymerase chain reaction machines, etc). For a biosafety level 3 facility, the equipment is stainless steel where possible, but there is a variety of materials, including electronics and other sensitive equipment. No corrosion has been experienced on any equipment or surface despite repeated decontaminations. Even the most sensitive equipment has not experienced any ill effects. As a test, when decontaminations were started, a low-cost laptop computer was moved from room to room to a Class III cabinet, decontaminating it every time in each chamber. After 35 repeated exposures, it was still functioning with no issues. It is still in use in one of the animal rooms and gets decontaminated along with the room. The Class III cabinet has not shown any traces of corrosion on the stainless-steel interior, nor has the stainless-steel inhalation exposure system kept inside the cabinet experienced any ill effects.