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Researchers safety

With the classification of four ‘Risk Groups’ of infective microorganisms and safety precautions in four ‘Bio-Safety Levels’, the biological safety and security issues are considered, standardized and handled. These criteria can be found in WHO Laboratory biosafety manual.

Generally, Escherichia coli TOP10 cells are used for cloning. They are derivatives of the non-pathogenic K-12 laboratory strains. In our lab, DH5alpha is used. The detailed information of the E. coli mutations can be found in E.coli wiki. The likelihood of a human getting ill from working with these bacteria is therefore low. The most probable route of transmission would occur by accidental aerosol formation or ingestion. The Environmental Protection Agency states that the K-12 strain (which both DH5-Alpha and JM109 are derivatives of) is poorly retained in the human gut, so the chance of it becoming pathogenic by mutations is low[1]. Mainly for these reasons, this organism is classified as Bio-Safety Level 1.

Aspergillus niger N593, and 872.11 are all derivatives of N400, which is the sequenced CBS strain. They are non-pathogenic industrial 'workhorses'. Additionally, the strains N593 and 872.11 have the pyrA auxotrophy marker. (see Fungi-related safety).

Public safety

In case of the accidental release of bacteria containing our BioBrick system, the physical conditions outside the lab are harsh to the E. coli strains that are used, so the bacteria would be unable to grow and the DNA of the BioBrick system will not be widely spread. By means of natural genetic transformation the DNA of the BioBrick system could be taken up by other bacteria.

Additionally, as expression of the plasmid would incur a significant metabolic burden on a (probably non-pathogenic) soil bacterium, and do little to increase its fitness, it is considered unlikely that the plasmid would be propagated. Unfortunately, it is not possible to know in advance what the actual effect of a natural transformation would be, but the odds seem to be in favour of it to cause little impact on the public and environment.

Environmental safety

There are several scenarios in which unintentional release of genetically modified material could take place. Labeling of the lab equipment and glassware used is necessary to prevent loss and improper waste disposal by a fellow researcher. The following problems are not that easy to prevent:

The air filtering system will have a hard time in keeping aerosols in the lab when a window gets broken. If this happens through a thunderstorm, an electricity break-down is likely and would increase the chance of release even more. Under Good Microbial Practice, though, the formation of aerosols is prevented as much as possible. Thus in total, the probability is relatively low.

All waste, containing genetically modified organisms, is required to be sterilized by autoclaving before disposal. Direct actions should be taken if it is discovered that an autoclave has been malfunctioning after the waste is discarded outside of the lab. To reduce the risk on environmental contamination it is necessary to check upon the autoclave’s functionality (by monitoring its operational temperature). If the aforementioned hazardous events do occur, they should be reported to the Minister of ‘Housing, Spatial Planning and the Environment’ and involved institutions to make the hazards undone as soon as possible.

The usage of antibiotic resistance markers increases the chance of spreading antibiotic resistance to pathogens. By conjugation, transduction or natural genetic transformation, DNA can be transferred between bacteria. There is a chance that the antibiotic resistance genes end up in a pathogenic bacterium which is not intrinsically resistant to antibiotics. The chance that these genes persist is low, since there is no direct evolutionary benefit for micro-organisms living outside the lab to take up the extraneous genes. The plasmids used in our lab can bear resistance markers for all different kinds of antibiotics such as Ampicillin, Chloramphenicol and Kanamycin. These are all resistance markers that are used in BSL-1 laboratories on a standard basis.

Risk and benefits

Generally, under Good Microbiological Practices the risk of working with the BioBrick system is rather small to the researchers. Despite the involved BioBrick system, the chassis in our project is considered to be safe to researchers, publics and environment. Even if the system was to be released into the surroundings, there is a low risk that it would enter a pathogenic host organism. However, there might be some risks involved if a person or organization has malicious intents by using this system as producers for toxins. In this case they would be able to use it to produce some secondary metabolites which have bad influences.

We believe that every new tool that is made could be used for malicious purposes, but by no means does this indicate that they should not be made. In our case, we think that the possible benefits of our system and the likelihood of it being used for those purposes outweigh the risk of it being used for evil purposes. Based on these considerations, we conclude that the potential benefits in successfully executing our project outweighs the estimated risks.

References

1. EPA (U.S. Environmental Protection Agency). (2011, January Monday). Escherichia coli K-12 TSCA Section 5(h)(4) Exemption: Final Decision Document. Retrieved July 2011, from Biotechnology Program under the Toxic Substances Control Act (TSCA):
http://www.epa.gov/biotech_rule/pubs/fra/fd004.htm