Team:OLS Canmore AB CA/Safety

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Safety

This year the Our Lady of the Snows Catholic Academy iGEM team has taken into consideration and put into action many different safety practices in order to keep everyone in our school, community, team and the world around us safe. With these considerations taken into account, we are confident that when our project is successful, it will benefit everyone. Our project will benefit those in municipalities who are currently dealing with hair buildups in wastewater treatment facilities by allowing a more efficient method to deal with clogs. Farmers in the poultry industry will also benefit from our construct because it will allow them to eliminate their feather waste but also to utilize the byproducts created for useful purposes. These benefits however, can only be applied if our construct is safe. If we continue to practice basic lab safety, lab safety education, the creation of a safe construct and thinking about future goals, we are confident that we will create a safe construct ready for proper implementation in the poultry industry and in wastewater treatment both locally, and on a global scale.

This year our safety concerns were focused around two major things. Firstly, our project could be, in the near future, released into municipal water systems or onto farm environments. Our second concern was that we attend a pre-K through twelve school, with our lab just off the main science lab, making the lab accessible to anyone when unlocked. These two things have meant that we must take safety into consideration both in and outside of a lab setting. Everyday we take simple precautions to make sure that us and everyone in our school remains safe. These important precautions are simple but very effective including wearing lab coats, goggles, gloves, locking doors of our lab at all times, washing our hands after lab work has been completed, and tying back hair while working in the lab. We sterilize all of our equipment before and after we use it and use 70% ethanol clean the counters in the lab. We have also undergone certain education in order to know how to be safe in the lab including; lab safety training, in which all iGEM team members and supervisors have learnt, Workplace Hazardous Materials Information System (WHMIS), material safety data sheets (MSDS), aseptic technique, proper sterilization techniques, and biosafety and wet-lab safety protocols. These safety precautions are not very complicated but very important when eliminating risks concerning our project, particularly when taking into consideration that some risks our project poses at the laboratory stage include: bacterial contamination; contact with harmful acids, nucleic acid stains, restriction enzymes, Keratinase; burns caused by a flame or warm glass or metal.

Lab work workshop at the University of Lethbridge

We have also taken into account some more complicated precautions having to do with the creation of our construct and the different forms of bacteria that we will use to build it. For the basic construction of our project we have planned to utilize an IPTG inducible promoter, but we plan to put a different promoter in the construct for implementation, as a way to allow for a control while testing within the lab in our assays we will perform. We plan on putting a different promoter in the construct, to make sure it is safe for implementation, as well as an efficient and cost-effective method, due to the cost of IPTG. We have also planned to implement a strong kill switch into our construct to allow for ultimate safety if ever the enzyme were to get into the environment or denature. The strain of E. Coli we are using is K-12, a genetically modified, non pathogenic bacterium which has a low probability of survival outside of a petri dish. Furthermore we do not plan to experiment with any other organisms besides our chassis E.coli, a lab strain that is not harmful to humans. We have grown our bacteria on plates that contain different antibiotics such as ampicillin and chloramphenicol, and placed a resistance to the antibiotic within the constructs to help ensure that our bacteria that grows is what is needed with our construct and stay safely inside the petri dish.

For our tests in the near future we will be performing both a qualitative and a quantitative assay in order to test the effectiveness of our project in more than just one way. These assays will be performed on both real hair and feathers meaning that we must take into consideration certain safety precautions. Many of the steps we will take to ensure safety will be our basic lab safety precautions, however some will be more complex such as learning to deal with the byproducts of the reaction, how to extract them safely to potentially be put towards a new product, and in which conditions we should perform this test to keep all students at our school safe from anything given off by our assays. We will maintain our safety standards while measuring in our assays to ensure our construct can be the best it can be.

Safe implementation is one of our teams largest concerns as many people in our community, as well as the surrounding area will be effected by our project after implementation due to it being designed for use in wastewater treatment plants, poultry farms, and at rendering plants. Because these are such big industries that effect people on a large scale, we must ensure our construct will not cause harm in anyway before we take it outside of the lab. We strive to achieve a safe and easy working construct when considering the implementation process and the precautions taken to do so safely for everyone. At the moment we are taking into consideration different ways to prevent leaks or exposure of our construct into the environment, water source, and general public. The environment is of huge importance to us because we live in a wildlife corridor outside a national park. We do not want our construct to leak into the ecosystem and cause potential harm, so containment is vital. In order to prevent leakage and contamination, we have been working on the creation of a containment chamber for our enzyme that will be safe and easy to implement on a mass scale. Through extensive research we have determined that a membrane bioreactor or an MBR would be the best way to safely contain the enzyme.

Bioreactor at our local wastewater treatment facility

Membrane Bioreactors are the safest option for a containment vessel in wastewater treatment facilities and on farms due to them being closed containers. Many self clean and will not break with ease (1). Membrane Bioreactors are the absolute best way to implement our construct in the ways of hair as they are the most effective at breaking down waste, have many positives and are the safest containment option. MBR’s are also safe for the environment and animals. Bioreactors of all kinds can be dangerous however with simple training which most people in wastewater treatment plants already have they are safe with little chance of injury, problems or death. The most common safety concerns are as follows:

Bioreactors can be explosive if incompatible chemicals are placed in or around bioreactors Bioreactors generate carbon dioxide if one is in an enclosed space with a bio reactor they can experience oxygen deprivation Clothing which is baggy can be easily caught in bioreactor or other equipment associated with bio reactors. Workers may be exposed to waste contaminants by inhalation, ingestion or absorption. Biological activity of the bioreactors may be enhanced with the addition of nutrients or other chemical agents. These agents may include nutrients, methanol, or other chemicals for pH adjustment (e.g. acids and bases). Workers may be exposed to these chemicals during their application either as a powder or in a liquid state. Bioreactors may expose workers to pathogenic microbes during operation and maintenance. However, exposure to these pathogens is usually not a significant concern unless the wastes being fed into the reactors contain pathogenic agents. If the bioreactors are equipped with open aerators, microbe-entrained mists may become airborne. Inhalation of pathogenic microbes may cause allergic reactions or illness. During sludge handling activities, workers' hands may be exposed to microbes and result in accidental ingestion of pathogenic material (1).

In conclusion our team has taken many different safety aspects into consideration when working on our project. The safety precautions are critical to our next steps when moving forward in the future with our project plan.

Citation: FRTR, Radiation Technology Screening Matrix Reference Guide http://www.frtr.gov/matrix2/health_safety/chapter_14.html

Contact us at alinaarvisais@isidore.redeemer.ab.ca or taliadixon@isidore.redeemer.ab.ca, or even send a message to our facebook page: https://www.facebook.com/OLeSsence