1. Safety and Security in Project Design 1.1 Organisms Used The E. coli strains we use are whitelist species and are a communal strain that has been used as a probiotic agent to treat gastrointestinal infections in humans since the early 1920s. If the E. coli is released into the environment, due to the weakening of its varieties, its survival ability has been greatly reduced, which will not cause ecological problems. Occasionally, however, probiotics may have adverse clinical effects, including septicemia. If both the microbiota and adaptive immunity are defective, translocation across the intestinal epithelium and dissemination of the probiotic E. coli strain may occur.
Furthermore, the Xanthomonas strains we used in the experiment are all level-4(the lowest) bacteria strains that can be used in education and experiments. Including Xanthomonas orzyae strain and
1.2 Experiment Design First, we have to ensure that xoo-sp2 will not present any infection to E.coil, Bacillus, Acetobacter xylostacter, and Roxella, which will be commonly used in our laboratory. According to the analysis, the xoo-sp2 phage will survive only a few hours in soil and causes no damage to human.
Second, considering the possible use of Xanthomonas in our project, we will ensure that it will not leak from the laboratory. The experiment will be conducted in the proper circumstance and will strictly follow the instructions given by our instructor. Besides, Xanthomonas is mainly transported by water, so we will sterilize all instruments and surplus fluid as we finish the experiment.
As for the chassis, the Xanthomonas oryzae strains we use are contaminated especially for plants in Oryza, and further cause rice bacterial leaf blight and rice bacterial leaf streak. However, based on our research, we already have a clear notion of the phenomenon that will present as Xoo bacteria leak from Petri dishes and methods to eliminate the Xoo. Therefore, we can set up a strain of rice in the lab and make sure it sterilizes frequently and fast.
Also, Xanthomonas oryzae bacteria itself and its metabolite will not cause damage to humans. Instead, Xanthomonas maltophilia, a different kind of Xanthomonas, can enter a human's respiratory tract and causes serious human respiratory inflammation. Thus, we will not consider harmful Xanthomonas like Xanthomonas maltophilia and focus on the Xanthomonas oryzae bacteria.
2.Safety and Security in Laboratory Work 2.1 Training Enough to equip team members who participate in wet-lab experiments with necessary knowledge and skills that ensure biosafety and biosecurity. The supervisor offers a lab instruction booklet and an online quiz to make sure every team member knows the basic lab operation. All team members are required to complete the rigorous examination related to biosafety and biosecurity before conducting wet-lab experiments. The exam contains 42 questions and covers every aspect of the lab activities. Members who fail to score over 80 percent are forbidden to enter our labs. We learn about lab access and rules (including appropriate clothing, eating, and drinking), responsible individuals (such as lab or departmental specialists or institutional biosafety officers), differences between biosafety levels, biosafety equipment (such as biosafety cabinets), good microbial technique (such as lab practices), disinfection and sterilization, emergency procedures, transport rules, physical biosecurity, personnel biosecurity, dual-use and experiments of concern, data biosecurity, and chemicals, fire, and electrical safety. 2.2 Lab Setting The biosafety level of our workplace is level 1. The labs in which all our wet-lab experiments are conducted were built under the supervision of experts and were approved by the Chinese government. Risk management tools including accident reporting (measures to record any accidents), personal protective equipment (including lab coats, gloves, eye protection, etc.), inventory control system (measures to track who has what materials and where they are), access controls (measures to control who can access your workspaces, or where materials are kept), waste management system (measures to make sure waste is not hazardous before it leaves your institution) cover our work. The officers provide all assistance in experiments requested by our members such as the usage of PCR machines, cleaning used instruments, and turning off the electricity and flames before leaving. Our supervisor researched a lot of the usage of our E. coli, and he is familiar with the experimental procedures and practices. 2.3 Containment When conducting wet-lab experiments, all team members are supervised by experienced lab experts, and the protocols used are seriously examined to ensure biosafety and biosecurity. All team members are required to dispose of waste appropriately. They strictly follow lab protocols and make every effort to prevent leaking. 3. Safety and Security in Human Practice 3.1 Surveys and Interviews We engage with communities to understand broader social concerns by talking with specialists in this area. All human subjects research conducted by teams is done in accordance with relevant laws and regulations, as well as any institutional rules or guidance, and above and beyond in demonstrating the commitments to responsible and ethical conduct. Our human subjects research is ethical and responsible for issues such as informed consent and privacy and data protection. Recorded verbal consent is made and specific permission from the cited individual to use their name has been obtained. 3.2 Real Application About the Xanthomonas oryzae, the bacteria itself and its metabolite won't cause damage to laboratory technician's skins. For example, xanthan gum, the Xanthomonas fermentation products. However, when inhale the Xanthomonas by accident, it will cause acute inflammatory disease of the lungs, especially using the Xanthomonas maltophilia to conduct experiments. So we will not use Xanthomonas maltophilia to minimize the potential healthy problems caused by our experiment.

Also, the xoo-sp2 phage we use is harmless to human body. We just need to pay attention on the disposal of liquid waste. If the phage delivery system releases to the environment without control, it can really cause a problem. The phage delivery system contains xanthomonas, and the leakage of that bacteria will be harmful to other crops and vegetations in the same area, thus causing severe decline of the production.

In addition, high concentration of bacteria in the phage delivery system will be a potential risk. In fact, it will lead to biological fermentation. Xanthan gum, the product of Xanthomonas fermentation, may not only affects the activity of our designed bacteria but also causes some digestive problems to human.