Difference between revisions of "Team:OLS Canmore/Safety"

SAFETY

The iGEM team of Our Lady of the Snows Catholic Academy has considered and implemented various safety practices in order to protect our students, staff, team, and world around us. As we have taken safety considerations into account, we are are confident that our final product will have an overall beneficial effect. Our project will prove to be beneficial for those in municipalities who are dealing with the buildup of hair in local wastewater treatment facilities, as a more efficient method to rid of these clogs would present itself through our construct. The poultry industry would also benefit from our project because it will allow for the elimination of feather waste, while also creating useful byproducts. However, these benefits are only applicable if our construct is safe. As we continue to practice safety in the lab and lab safety education, we are optimistic that we will create a safe construct that is ready for proper implementation in wastewater treatment facilities as well as in the poultry industry, both on a local and global scale.

Two main safety concerns that have revolved around our project since the beginning are that our project, in the future, would be released into municipal water systems or farm environments, as well as the fact that our school ranges from grades pre-K to 12. In addition to our own students, our school also neighbours another school, who on occasion utilizes our facilities as well. Our lab is situated just off the main science lab and when unlocked, it is accessible to all students and staff members. Due to these two important factors, it is vital that we take safety into consideration, both in and out of a lab setting. The precautions we take for the safety of all are simple, yet very effective. These precautions include wearing protective gear (such as lab coats, safety goggles, and gloves), locking the doors of the lab at all times, washing our hands after lab work has been done, and tying hair back to avoid contamination. All of our equipment is sterilized prior to working in the lab as well as after, and 70% ethanol is used to clean our lab counters after use. We also have additional safety protocols that we practice, as we have limited waste disposal methods in our lab and due to a considerably low budget, the majority of our equipment is either second-hand or Do-It-Yourself. Previously, our team had undergone certain education to become knowledgeable on lab safety including; lab safety training, Workplace Hazardous Materials Information System (WHMIS), Material Safety Data Sheets (MSDS), aseptic technique, proper sterilization techniques, and biosafety and wet-lab safety protocols. Though not complex, these safety precautions are crucial to the elimination of risks associated with our project. Said risks that our project currently poses include: bacterial contamination, contact with acids, nucleic acid stains, restriction enzymes, or Keratinase, as well as burns that can be caused by a flame, warm glass, or metal.

Gloves are a necessary safety precaution, as well as a flame for decontamination. Talk about how we using one of the safest nucleic acid stains for dying gels (redsafe). As examples of substitutions to increase safety. There are three steps for safety and these include administration (ppe, we don’t really use that many of these because nothing is super dangerous), substitution (ex. Redsafe vs ethidium bromide), and engineering (ex. fumehood) precautions. Although ethidium bromide is a highly sensitive stain that produces good results in agarose gels, it is notorious for its toxicity.

Though we take basic lab safety precautions with great seriousness, we must also consider some complicated precautions in regards to the creation of our construct and the form of bacteria we are utilizing. For the basic construction of our project, we have used an IPTG inducible promoter, however during actual implementation, we plan to replace this with a different promoter that is more suited for a full-scale model. By putting a different promoter in our construct, we are making it safe for the implementation and are creating a cost-effective and efficient method, as IPTG is very costly and will not be ideal for wide-scale implementation. We have also decided to implement a strong kill switch into our construct, allowing for safety in the case that the enzyme were to negatively affect the environment or denature. When we think about the application of our project, there are many variables to consider, and addressing all of these variables will require an extensive amount of planning, research, and years of development.

Chassis-wise, we are utilizing the K-12 strain of E. Coli, which is a genetically modified bacterium that is non-pathogenic. This strain of bacterium has a low probability of survival outside of lab conditions. We do not plan to experiment with anything other than our chassis, E. Coli, as it poses no threat to human life. Our team has grown our bacteria on plates that contain antibiotics such as ampicillin and chloramphenicol. To ensure that our bacteria grows to be what is needed and stays safely inside the petri dish, we have also placed a resistance to these antibiotics within our construct.

Our team has performed two assays in order to test the effectiveness of our project: a dry hair assay and a skim milk plate assay. These assays were performed on real hair and feathers, meaning we had to take certain safety precautions into consideration. Basic lab safety precautions are a given, however some precautions are more complex, such as learning how to deal with the byproducts of the reaction, how to extract them safely and securely, and in which conditions we should perform these tests in order to keep our school safe from any product that may be given off by our assays. We have maintained our safety standards while completing these assays and the safety of those around us is of main priority.

The safe implementation of our project is a large concern for our team, as our community and the area surrounding it will be directly affected due to our construct being designed to be applied in wastewater treatment facilities, as well as poultry farms and rendering plants. Due to the fact that these companies are vital and affect individuals on a large scale, it is of great importance that our project will not cause harm in any way. Thus, we must make sure it is completely safe before removing it from lab conditions. Currently, we are delving into different methods to prevent leaking and exposure of our construct into the environment, general water source, and the general public. Being situated in a wildlife corridor outside of a national park makes the safety of the environment a priority of our team. Containment is necessary as we do not want to cause potential harm to our animals or ecosystem as a whole. Through extensive research, we have determined the best way to contain our enzyme, in a way that is safe and easy to implement on a mass scale: a Membrane Bioreactor (MBR).

A Membrane Bioreactor is an apparatus in which a biological process or reaction is carried out. To put simply, they are closed containers that are used on an industrial scale, used commonly in most wastewater treatment plants. Many of them are self-cleaning and are difficult to break. Utilizing a bioreactor would allow for safe and easy integration of our system into an already-existing system and this stable, enclosed environment would prevent uncontrollable rates of growth and mortality of the bacteria, something that poses potential hazards in the industrial application of our project. Our bacteria would thrive at maintained optimal conditions and would degrade hair and feathers in a controlled fashion. Leaking would also be prevented, eliminating the risk of harming the environment and community. Certain kinds of bioreactors would completely remove the bacteria from water before reintroducing it to the environment and eventually, the general public. However, like everything, bioreactors of all kinds can pose threats. If an individual has the proper training required to operate a bioreactor, the chances of injury, problems, or death, are reduced. The most common safety concerns include:

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 bioreactor they can experience oxygen deprivation. (Only in certain situations where CO2 is produced) 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). This year, our team is in the beginning stage of building our own small-scale bioreactor for experimentation purposes. Because we are only in the construction phase at this time, we have not yet had to implement important safety precautions regarding the bioreactor. This will change however, once our bioreactor is ready for the implementation of our construct.

To conclude, our team has taken many different safety factors into consideration as we move forward with our project. We ensure that safety precautions are met in and out of the lab space, as they are critical to the current and next steps of our project.