Team:Alberta/SafetyForm

Summary of Safety Form

There are no significant risks, as the biological parts are not reasonably able to cause any health issues to team members.

All the components of our project are quite standard in molecular biology labs and are not associated with significant risks. Our microorganisms, biological parts, laboratory equipment and chemicals are mostly comparably benign. The strains which our team intends to use and has used thus far are harmless (nonpathogenic and non-toxigenic). The biological parts we are using consist of color, antibiotic-resistance (Chloramphenicol, Kanamycin, Ampicillin, Tetracycline) and various promoters. In terms of chemicals, the myriad of flammable solvents in the lab pose a notable risk, and in addition the ethidium bromide (EtBr) is a dangerous toxin and mutagen. EtBr is used to visualize DNA by soaking agarose gels in a solution of the compound. EtBr works by intercalating between bases in the DNA’s helical structure, where is it able to fluoresce much more brightly than it does while in solution. It is generally established that intercalating agents are mutagens, and that mutagens are carcinogens and for this reason EtBr is treated with a high level of caution in our lab. Experiments have shown that EtBr is able to cause DNA damage in human cell lines so this carcinogenic potential is not just theoretical. To mitigate risks, relevant microbiological techniques are consistently used in the lab and gloves are worn. Other precautions for EtBr include the labelling of designated containers for EtBr gels and EtBr waste, and the reuse of EtBr solution to minimize waste.

The most dangerous laboratory equipments we use are a transilluminator, gel apparatus, and the Bunsen burner. The risks posed by exposure to UV radiation, electric shocks and open flames can be easily controlled with proper safety training and protective equipment. A thick glass cover has been added to one UV visualizer in the lab to protect eyes when visualizing gels. Usually, a different UV visualizer is used for lab purposes because it photographs the gel without any UV exposure to eyes at all. Students are taught how to properly use equipment, dispose of wastes, autoclave used glassware and biologicallycontaminated waste, disinfect counters and know when to wear safety protection. Proper pipetting protocol is always used.

The DH5-α strain with which our group is working (which is a derivative of K-12) is non-pathogenic and non-toxigenic, so it poses no risk to public health and safety.

A public concern is that the plasmids containing antibiotic resistance genes could be released into the environment. Since our project uses antibiotic resistance genes as selection tools, we ensure that these genes are properly disposed of by autoclaving.

In evaluating the risks, there is a lower chance of gene uptake by wild bacteria compared to our labstrain bacteria that have perforated membranes. In the case that wild bacteria did uptake the antibiotic resistance gene, that gene would eventually be lost in the population without exposure to the antibiotic, because it is then a disadvantage for the bacteria to carry the unused plasmids and use up energy to express them. Evidence for this was observed in multiple cases of measured declines in particular antibiotic resistances after the human population limited the antibiotic usage (Klotins, 2005).

The bacteria that we use in the laboratory are not well suited to survival in the environment and even assuming they were able to do so, they do not have any destructive potential. The standard and well-characterized biobytes we are using do not have the potential to combine in dangerous ways. As with many other synthesized DNA constructs, we have found that our plasmids are lost from the bacteria over time (if plated without antibiotic present), due to the expression of colors being an unproductive strain on the cell’s resources. Consequently, there is little risk of our microorganisms spreading. Even if our E. coli strains survived and replicated in the wild, they are highly unlikely to cause harm to any humans or animals as risk level one microorganisms. The microorganisms we are creating in the lab would have less potential for malicious misuse than any bacteria normally found in the environment.

Although there is no intentional release to the environment and people, contamination is minimized by protective equipment (gloves) and disinfection. However, the microbe strain cannot survive outside the lab environment because it is engineered to be weak and dependent on a lab nutrient mix (Luria Broth, LB agar). In the improbable case that the microbe evolved (ex. By uptaking genes) to be able to synthesize one of the essential amino acids, it still would be deficient in multiple other ways.

Future risks from Commercialism

Answering this question accurately is something of a complex endeavor, as one of the key aspects of our project is that it is essentially a foundational advance. As such there are probably many approaches a focused group of minds might be able to develop using some of the principle and methods we have developed. However, even given the very open-ended nature of our project’s potential future, it is difficult to envision scenarios in which significant dangers are encountered. It is possible that in the distant future our methods might be modified and used with organisms and/or genes that entail greater risks than the non-pathogenic E. coli we have chosen to use. Even in this instance, the risks to personal, environmental and public safety are equal to the risks posed by the organisms and/or genes used and not modulated upward by the methods of our project in any conceivable way.

Laboratory safety training is given to undergraduate students in many of their laboratory courses. Specific to bio-safety, three of our team members have completed a comprehensive biological laboratory safety course in their Methods of Modern Biochemistry course (BIOCH 401) This course includes general biological, chemical, and WHMIS safety training). All students including the high school students were given on-site training for the specific techniques they would be using in the laboratory.

Biosafety Committee

As is the case with most universities, there is a local Biosafety Committee on the university of Alberta Campus that ensures research labs are following the safety protocols put in place by the national government. The University of Alberta Biosafety Committee is a division of the Environmental Health and Safety Faculty. This Committee oversees the handling of biohazardous materials and/or techniques, defined as “materials of biological origin or synthetic material which mimic biological entities and may induce adverse conditions to humans, other animals, plants, or the environment”. With respect to relevant national and international guidelines, our non-virulent strains of E. coli are considered Biosafety level 1 and are of limited potential hazard. Within our lab, standard practices are used, including but not limited to wearing gloves and sterilizing with bleach and/or ethanol and/or autoclaving. University policy states that the Biosafety committee need only be contacted when risk group level 2 organisms are to be worked with, or for other special concerns, and as such there is currently no need to discuss the project details with them to obtain their approval. However, the EHS provides documents on laboratory guidelines and university-specific ethidium bromide disposal (EHS, 2011), which have been reviewed by the team.

According to the WHO bio-safety manuscript, the living organisms with which this year’s University of Alberta iGEM team has worked (and will work with) all fall within the category of Risk Level 1, which the manuscript goes on to define as “unlikely to cause human or animal disease”. Since the strains which our team intends to use and has used thus far are harmless (non-pathogenic and non-toxigenic), they all fall into this category and as such, no major safety issues are raised by the biological components of our project.

References:

• Ethidium bromide – fact sheet. November 2011. Environmental Health & Safety. Retrieved August 9, 2013, from www.ehs.ualberta.ca/EHSDivisions/~/media/Environmental%20Health%20and%20Safety/Documents/ Chemical%20Safety/FactSheet_Ethidium_Bromide_-_2011.pdf
• Klotins, K. (n.d.). Antibiotic use for growth improvement. Ontario Ministry of Rural Affairs / Ministry of Agriculture and Food. Retrieved August 2, 2013, from http://www.omafra.gov.on.ca/english/livestock/animalcare/amr/facts/05-041.htm
• http://www.ehs.ualberta.ca/EHSDivisions/~/media/Environmental%20Health%20and%20Safety/ Documents/Manuals%20and%20Documents/Biosafety_Manual_Aug__03__2_.ashx
  (Found also from: http://www.ehs.ualberta.ca/EHSDivisions/Biosafety.aspx)
• http://canadianbiosafetystandards.collaboration.gc.ca/cbsg-nldcb/index-eng.php?page=1 and
• http://www.phac-aspc.gc.ca/publicat/lbg-ldmbl-04/.