Team:Stanford-Brown/HumanPractices/Safety

Safety

At NASA Ames Research Center, we are part of the Education Associates Program (EAP) which requires all interns to undergo thorough lab safety training, including biosafety. This includes a four hour safety “bootcamp” which covers biological containment protocols, waste disposal, and handling of hazardous materials. All are required to take a test following this session which certifies safe lab work under NASA Ames Guidelines. All members have been received credit for this training.

NASA Ames Bio safety guideines: http://server-mpo.arc.nasa.gov/Services/CDMSDocs/Centers/ARC/Dirs/APR/APR8800.3C6.html

NASA General Safety Guidelines:http://server-mpo.arc.nasa.gov/Services/CDMSDocs/Centers/ARC/Dirs/APR/APR1700.1.html

Stanford APB website: http://www.stanford.edu/dept/EHS/prod/researchlab/bio/

1. Would any of your project ideas raise safety issues in terms of:

-researcher safety?

It is imperative to first note that our project utilizes Biosafety Level 1 organisms (BL1), and as such, our project is harmless to researchers. The organisms we utilize: K-12 Escherichia coli, Bacillus subtilis, and Deinococcus radiodurans are non-infectious and non-pathogenic, although the Homo sapiens present in the lab may present some hazard. However, given the scope of our projects, each component requires careful consideration regarding safety:

Venus: The successful aerosolization of bacteria would increase the potential for contamination of the labspace in general. However, all attempts to aerosolize bacteria occurs within the isolated chamber of the adapted Millikan apparatus, and, we reiterate, only K-12 E. coli will be used for this experiment, so this experiment should be non-problematic from a safety standpoint.

Biomining: We are engineering bacteria to express silica-degrading carbonic anhydrase. While this does not pose direct threat to humans, it could compromise the integrity of glass labware, possibly leading to laboratory accidents or property loss. Thus, for testing carbonic anhydrase, we are particular to use plastic and other non-glass equipment when possible.

Hell Cell: None of the bacterial strains or genes we are working with pose any threat to researcher safety. However, since our assays involve testing with extreme conditions such as high levels of radiation, acidity, and basicity, we are careful to use corresponding PPE (Personal Protective Equipment) and to correctly use machinery (i.e. lowering the hood when applying radiation) to minimize risks.

-public safety?

Bioaerosols in confined indoor spaces can pose risk for public health. Tringe et. al. have shown that indoor airborne environments house microbiota adapted to indoor spaces and human carriers. Thus, aerosolized bacteria might not be only confined to the lab, but might also escape to public and domestic indoor spaces visited by lab workers. Glass and silica constitute major elements of modern societal infrastructure, from windows, electronics, and optic cables. Thus, the escape of a silica-degrading microbe could have disastrous consequences for human urban environments. It’d make a cool movie though. Thankfully, the K12 E. coli we are using to degrade the silica is not likely to survive outside of its LB home, and can easily be eliminated with ethanol.

-Environmental safety?

The atmosphere constitutes a spatially, temporally diverse microbial environment, with possible effects on aquatic, terrestrial, and human health (Amato et al.). These processes range from ice and cloud nucleation to plant pathogens (Delort et al.) Furthermore, atmospheric currents have the potential for dispersal of bioaerosols. Thus, the release of genetically engineered aerosolized bacteria could have widespread consequences. The Hell Cell looks to push life to the extremes by overcoming evolutionary metabolic and physical limitations. While it may seem that bacteria from the Hell Cell could possibly outcompete native microbes in extreme niches, the ecological niche has not been expanded enough to make a significant difference in the bacteria’s survivability outside the lab.

However, the Hell Cell ultimately seeks extraterrestrial applications, and this raises real concerns surrounding forward and back contamination. Forward contamination is accidentally taking microbia from Earth to other planets, whereas back contamination is the process of taking microbia from other planets back to Earth. Both processes have the potential to seriously disrupt recipient environments. First of all, it is important to note that Planetary Protection Policies currently address the issue of microbial contamination of extraterrestrial environments and impose strict control on what can and cannot be released in space. These policies are enacted in order to avoid contamination and preserve other worlds in their natural states and vary for each celestial body (e.g. we could not test our constructs on Mars, which is protected from any terrestrial contamination). However, to protect against the event that one of our projects causes contamination in space or on Earth, we would engineer inducible kill switches into these bacteria that could annihilate them if need be.

To learn more about Planetary Protection Policies: http://planetaryprotection.nasa.gov/

2. Do any BioBricks raise safety issues?

No BioBricks we are creating raise significant safety issues: none of the proteins our genes code for increase the pathogenicity of the bacteria nor confer potentially hazardous functions. The lone exception is carbonic anhydrase, which can degenerate silica; this will be noted in our documentation of the BioBrick. When handling strains containing this gene, we take care to use equipment that is not made out of glass in order to prevent possible accidents.

Outside of this exception: Venus Life deals with isolating promoters that do not inherently carry potential hazard, the engineered flagellin Biomining is working with has no deleterious effects, and the proteins expressed in the Hell Cell project offer no major safety issues.

On a larger scale, the suite of BioBricks combined in the Hell Cell project may offer environmental hazards, as the engineered organism would potentially disrupt existing ecologies and be difficult to contain; however, the ecological niche has not been expanded enough to make a difference in the bacteria’s survivability outside the lab, and we have taken care to ensure that certain chemicals (e.g. ethyl alcohol) is still fatal to engineered microbes. In other words, given the nature of our project and safeguards we have implemented, none of our BioBricks pose a safety hazard to individuals nor the environment.

3. Is there a local biosafety group, committee, or review board at your institution?

-If yes, what does your local biosafety group think about your project?

All biological research and lab work was conducted either at NASA Ames Research Center or Stanford University. At the Stanford the biosafety authority is the Administrative Panel on Biosafety (APB). Any work using biological agents at BL-2 (Biosafety Level 2) or greater, and/or any work non-exempt from NIH rDNA guidelines are required to have APB approval prior to commencement. Our work only involves non-pathogenic strains of bacteria, which constitutes only Biosafety Level 1, and is also exempt from NIH guidelines under exemption clause (III-F-6). Thus this project satisfies the standard for safe practice at Stanford University. This is confirmed by team member Gabriel Ben-Dor, who last year was nominated to the Stanford’s Administrative Panel of Biosafety.

4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

Implementation of the kill switch system mentioned earlier could be useful for future iGEM competitions. While it is particularly relevant for our project in that it could offer prevention of possible forward or backward contamination, it is a significant safety consideration. iGEM and synthetic biology may benefit from engineering a kill switch into standardized plasmids (e.g. pSB1C3).