1 Introduction
As a synthetic biology team, we clearly recognize the significance of the safety of our project and prioritizes it before any other parts or results of our experiment. Our experiments used the common genetic engineering model strains - Escherichia coli and Bacillus subtilis, and all our experiments were conducted under the premise that the environment, equipment and operation were completely safe, while all our experiments were conducted under the reassurance that the environment, equipment, and operations are completely safe. Furthermore, we have ensured that our product would have no safety issues once entering the market. We have organized our safety page into four sections: general safety, experimental safety, project related safety, and project application safety.
2 General Safety
2.1 Behaviors
For this year’s project, our team strictly follows the regulations, which are classified as biosafety level (BSL) 1. All the team members have received detailed lab trainings and are required to achieve a merit on the exam of lab safety before entering the lab. When our team is in the lab, participants’ behaviors are carefully regulated.
The laboratory requirements include but are not limited to the rules listed below:
a. It is forbidden to turn the pipette tips, conical flasks, test tubes and other containers toward yourself and others’ faces to avoid liquid splashing
b. Before starting the experiments, check the experimental environment and reagents that need to be used in advanced. If it involves the use of odorous reagents (such as chloroform, isopropanol, etc.) you need to wear a mask in time, and ensure that the experimental space is ventilated and kept air flowing.
c. Experimental equipment, pipettes, and various reagents on the contaminated area (operating table for protein electrophoresis and nucleic acid electrophoresis) should not be taken to other areas. Gloves that have been in contact with this area should be thrown away directly, and other experiments should not be directly touched area.
d. Wear goggles and protective gloves before using the liquid nitrogen tank and place the operating procedures next to the liquid nitrogen tank.
e. The waste liquid (not exceeding 85% of the volume) and empty reagent bottles are labeled with orange waste labels and placed under the warning sign (big triangle skull logo).
f. The instructor of the biological project shall prepare a safety inspection log, and timely update and inspect the fire-extinguishing facilities of the laboratory, such as fire extinguishers.
2.2 Prevention (First aid kit)
The first aid kits are kept readily accessible at our laboratory, with all the required materials, medicine, and equipment in them. The medicines within the first aid kits are replaced regularly by the supervisors in the lab to maintain their effectiveness. All the materials are reassured to be effective in a timely manner just in case of an urgent situation.
2.3 Further prevention
There is also equipment for safety prevention placed around the lab, e.g., eye washer, fire extinguisher, blankets, lab coats, and goggles. All members in our team were educated and trained on the proper usage and circumstances. All prevention above can reassure that experimental accidents do not occur, and even if such accidents happen, every individual in the lab has the ability to handle the emergencies.
In order to prevent cross-contamination, our team made sure the experiments were done in aseptic technique when necessary. The laboratory equipment and material that has an aseptic requirement or environment are also guaranteed to be properly used.
3 Experimental Safety
3.1 General device usage
All of the devices that are common in a biological laboratory were taken good care of and used for suitable purposes. Our commonly used instruments like refrigerator, glass vessels, centrifuges and ice maker are all used with proper precautions, knowledge, and operation.
Visit the appendix below for more details about instruction for operating devices.
4 Project Related Safety
4.1 Clarifications
a. Throughout our project, there was no animal usage at all.
b. No special bacteria with potential dangers were used.
c. There is an extremely low potential risk of creating super bacteria. All the antibiotics were applied carefully, and we selected the antibiotics cautiously, making sure that even if experimental accidents occur, the risk of creating super bacteria is low.
4.2 Survey Participant Privacy Protection Clarification
a. Before every single survey or interview applied for our PE/HP, we will ask all the participants’ agreement to collect relevant data.
b. All recordings appeared in the wiki and all recordings we collected has been created with the interviewee’s agreement. If some interviewees preferred to not show certain body characteristics, our team members will strictly follow the interviewees’ preference.
c. We have also set promises to all interviewees and survey participants that all data we collected will not sent and appeared in other place rather than between our team members.
d. We also promise that the usage of all of our responses we collected will only be for our wiki creation, data analysis and project related usage, no more.
e. During data collection, if not has specific usage or specific people who has significant impact on our project, the interviewee’s personal privacy information will not be even asked during the collection.
5 Project Application Safety
5.1 Application
a. There are three possible pathways for our experiment product, Fengycin, to enter the market as our final product, a combination of deodorant and perfume. The first way is to be publicly sold after factories ferment Fengycin and mix it into solutions that are part of the final product. The second way is to be produced from our designed device: it will be a small equipment that loads components including Bacillus subtilis, cultures, filters, and then Fengycins were extracted from Bacillus subtilis inside the device. The third way is a plan that can be applied once regulations on biosafety are loosened: B. subtilis will be put into hydrogel, the latter allowing the malodorous molecular 3M3SH being assorbed by Bacillus subtilis inside, and fengycins to pass through and inhibit the quorum sensing of Staphylococcus spp., which should be responsible for body odor. Also, a suicide mechanism of the Bacillus subtilis is needed because the principle of DO NOT RELEASE should be put first in consideration of safety to both human and environment. make physical contact human skin to inhibit body malodor.
b. For the first way, safety is ensured as long as the factory employees properly follow standard procedures of Fengycin fermentation process. For the second way, the device has the extraction process taking place inside it, no direct contact with human body is made and the only possible outcoming product is Fengycin. For the third way, the designed structure of hydrogel allows nothing but Fengycin-size molecules to pass through the gaps between hydrogel molecules and make direct contact with human skin, ensuring living B. subtilis will directly contact human body and cause any potential dangers that will be discussed below.
5.2 Potential dangers
a. There are no major potential dangers led by our product.
b. E. coli and B. subtillis, the classic engineering model strains, were applied for genetic engineering and fermentation production. They do low harm to human body, as long as we properly operated. One of the greatest risk of using E. coli is the accidental ingestion of it into human body causing symptoms like diarrhea, while Bacillus subtillis—according to research—appears no hazard towards human body. Besides, proper disposal of bacteira in the fermentation broth should be given more attention otherwise the environment would be polluted.
c. When it occurs to experiments of pathogens, the experiment should be conducted in the laboratory of p2 biosafety level in case any leakage possibilities. In our experimental design, we detected of biofilm formation of Staphylococcus spp. to verify the effectiveness of our fengycins product. And after contacting with Doctor Zheng, the experiment was carried out in the laboratory in Nanshan Hospital.
d. Another potential risk is the use of SYBR (Synergy Brands, Incorporated) DNA dye during the DNA measuring process which, although relatively safe already, can possibly cause illness to any wound on human body.
5.3 Potential dangers’ prevention
In order to prevent the potential risk, we have adopted some prevention methods. If have the chance to publish our project into the market, above than our project, we will investigate a bacterial suicide system that will help to trigger the suicide mechanism of fengycins, and Staphylococcus spp. further causes to danger to the local environment after the product been used. Also, since we have a certain progress in the investigation of the suicidal system, we believe we have the potential to achieve such progress. This would allow our project to be more environmentally friendly also beneficial to the local trash classification system.
Furthermore, public education on the proper usage of the devices could also assist the process of prevention. By warning the public on such dangers when using the product, the targeted audience (citizens) would be cautious of the forbidden operations when having contacts with the device.
Hence, such potential risk could be prevented.
