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Integrated Human Practice
1 The Cause of Superbugs
Through research, we found out, around 1.2 million people's immediate cause of death is antibiotic-resistant in 2019. There is even an estimation that suggests 10 million people around the world will die each year due to antibiotic resistance bacterial infection in the future. To find out the reason, we designed a questionnaire based on how familiar people are with the knowledge of antibiotics and superbugs. From that, we found out that most people (ranging from teenagers to adults) were fairly well informed about the knowledge, with more than 60% of the participants giving the correct answer. Moreover, we had an interview with a doctor. The doctor suggests that the main cause is the abuse of antibiotics. However, pharmacies nowadays have very strict regulations about prescribing antibiotics. Therefore, animal husbandry is the industry whose causing more superbugs. In animal husbandry, sulfadiazine (SDZ) a type of antibiotic which is cheap and effective in killing bacteria is widely used. Therefore, we decided to focus on research and develop a product that could degrade SDZ.
Fig 1. Wrong or correct answer percentage to the question of overuse of antibiotics that do not kill
Fig 2. Wrong or correct answer percentage to the question of abuse of antibiotics would decrease human body immunity
Fig 3. Wrong or correct answer percentage to question abuse antibiotic would cause superbugs
Fig 4. Percentages that consider superbugs to be more pathogenic, infectious, lethal, and antibiotic-resistant
Fig 5. Percentages that think when antibiotics should be used
1.1 Doctor's Interview
Interviewee Introduction:
Yanbin Peng. Head of surgery in Shenzhen Peking University Hospital.
The doctor suggests that the main cause of superbugs is abusement of antibiotics. The health department recognized the problem and have set very strict regulation for prescribing antibiotic. We learned that: 1. Clinicians should provide the detection rate of pathogenic specimens before providing antibiotics, and rationally select drugs according to the results of bacterial culture and drug sensitivity test. 2. Avoid combination medication without reference to land acquisition. 3. Try to choose anti-infective drugs with less toxicity and side effects, prescribe drugs according to the blood concentration, and try to reduce the dependence on antibiotics. Although hospitals are restricted by different rules for prescribing antibiotics nowadays, the number of superbugs causing deaths is still increasing. For this situation, the doctor deduce that the answer could be found in animal husbandry. Due to the low level of knowledge farmers have and want to make a large amount of profit, they chose to abuse cheap and effective SDZ to avoid livestock from getting sick. This speculation was confirmed. After the meeting, our team found out that in China, livestock that has been tested as potentially hazardous to human or animal health requires harmless disposal through research. Apart from that, the doctor also raised a social problem that he found in his daily work. Due to the strict control of prescribing antibiotics in hospitals, patients cannot be randomly prescribed antibiotics. Therefore, the recovery time will take longer. This causes doubt in the patient toward the doctor's profession and may refuse to follow medical supervision. Therefore, the doctor suggests we to educate people about the dangers of the overuse of antibiotics.
Picture 1. Live interview with Doctor Yanbin Peng
2 Original design
In the beginning, we plan to feed livestock with the recombinant strain, so we chose the probiotic, E. Coli Nissle 1917 (EcN). And chose Laccase 6 (Lacc6) from Pleurotus ostreatus HAUCC 162. However, the experiment result is unexpectedly bad. The recombinant strain is killed before fully functioning.
3 Interview with professor
Interviewee Introduction:
Zekuan Yu. Associate professor of the Department of Biomedical Engineering, Fudan University.
After summarizing, we realized that the main problem arose in the application of the method of intercellular expression. Using this technique may cause the recombinant strain to have low efficiency in degrading SDZ. Therefore, we had an interview with a professor. We showed him our project design and the experimental data that we collected. We were told that the reason for the low degrading efficiency is that the SDZ has to enter the recombinant strain before being degraded. During this process, the recombinant strain may be killed by the antibiotic, causing lacc6 unable to function.
After noticing the problem, we asked the professor for solutions. In the solutions suggested by him, we chose the technique of extracellular expression. The advantage of this technique is that it eliminates the need for SDZ to enter the recombinant strain to trigger degradation. Antibiotics interact directly with lacc6, which is immobilized outside the cell membrane. This increases the lifespan and rate of survival of the strain. Moreover, lacc6 can effectively contact SDZ outside the cell to improve degradation efficiency.
We learned a lot from this interview. With the professor's guidance, we redesigned our recombinant strain. The correctness of this decision is also be confirmed in future experiments.
Picture 2. Live Interview with Professor Zekuan Yu
4 Redesign
After we decided to use the extracellular expression technology, we discovered a new problem. How to fix Lacc6 on the surface of EcN? By chance, we found two other teams that are also planning to use the extracellular expression technique. BFSU-ICUnited and LZU-hS-China-A. After communication, we found out that the other two teams were faced with similar problems as we were. We decided to help each other to solve the problem. First, we decided to review articles based on cellular expression technology and then share them. We analyzed the advantages and disadvantages of INP and other anchor proteins and compared them. After a series of analyses, we found that Ice Nucleation Protein (INP) meets the requirements of our design. After confirming with the instructor, the three teams decided to use INP. This collaboration is the beginning of a longer-term collaboration between our three teams in the future.
Picture 3. Discussion about the pros of different anchor proteins with teams BFSU-ICUnited and LZU-hS-China-A
5 Interview with farmer
Interviewee Introduction:
Mr. Xin Ding. Owner of the farm.
After the previous modification, the degradation efficiency of SDZ by the recombinant strain was greatly improved. So we tried to productize the recombinant strain into the animal husbandry market. The farmer is our targeted customer. Therefore, we decided to have an interview with him. Asking for feedback on our products. To our surprise, he said he wouldn't use our current product. Because he thought the recombinant strain would cancel out the function of the SDZ in the livestock's body. Meaning to eliminating the function of SDZ killing bacteria and getting rid of the plague.
At the same time, we are also aware of other problems with the original feeding method. Our products have not yet undergone any safety tests and experiments. Feeding engineered strains to animals could raise several safety concerns. Considering the safety of the animals and the potential risks that our strain might have, and the unethical of experimenting on animals we decided to avoid animal testing.
Picture 4. Interview with Mr. Xin Ding
6 The Second Revision of Design
Since feeding livestock with recombinant strain was not feasible, our team decided to sprinkle the recombinant strain in the form of freeze-dried powder onto livestock manure, which has large amounts of SDZ residue. This solution can solve the problems raised by the farmer without affecting the physiological health of the animals. On top of that, this scheme can achieve the same rate of degradation as before.
Fig .Getting our engineered strain in contract with SDZ concentrated feces.
To motivate farmers to use our products, we have designed an additional product: a septic tank. The primary function of the septic tank is to provide the necessary place for SDZ degradation. Second, it can collect the biogas produced by the fermentation of the recombinant strain and store the treated SDZ-free manure. Biogas can be turned into electricity to run the septic tank and SDZ-free manure is an excellent fertilizer. Given the potential hazards of the recombinant strain evolving into a superbug, the septic tank has a corresponding sterilization function.
7 Second Interview With Farmer
After the improvements, we interviewed the farmer again. This time, we started by telling the farmer why we designed the product, which pointed out that the misuse of antibiotics in animal husbandry has led to many deaths and serious environmental pollution. Then we pointed out the advantages of our product, which is different from the existing degradation methods, it will not lead to secondary environmental pollution, and can also create greater benefits in the case of degradation of SDZ. This is because we not only designed the freeze-dried powder, but we also designed the septic tank. The method of using the septic tank is very simple. You only need to put livestock manure and free-dried powder together into the septic tank. Then press the start button. In addition to the benefits of simplicity, it can also turn treated manure into fertilizer and recycle biogas for electricity generation. This can help reduce some of the farmer's operating costs. And, of course, we also consider the problem of biosecurity. So we have set ultraviolet light inside the septic tank, which is used to kill the recombinant strain quickly so that it doesn't accidentally get into the environment and cause superbugs. After learning about our improvements, the farmer expressed his great willingness to support and buy our freeze-dried powder and use the septic tank. Because he thought it would not only make it easier to dispose of livestock manure, but it would also bring economic benefit and we also took people's health into account.
Picture 5. Interview with Mr. Xin Ding
8 Business
To solve the problem of superbugs endangering human health and polluting the environment, we decided to mass-produce freeze-dried powder. each box (5G *10 bags). After considering the cost of strain, packaging technology, packaging materials, labor, and label testing, we decided to set the selling price at 0.65dollar/ box. Selling freeze-dried powder at this price can not only make profits to support our partial improvement of freeze-dried powder in the future, but also reduce competitors as it's a low price compared to others.
In addition, to increase customers' demand for our product, our team released two popular science articles about superbugs on our official account. These include educating the public about what superbugs are, what serious problems can arise from antibiotic resistance caused by superbugs, and calling for compliance with medical supervision. Because the online world is now so widespread, even people in rural areas (farmers) can learn science easily. In this way, we can make people aware of the importance of reducing SDZ and therefore increase farmers' purchase of our product.
Reference
【1】 University of Oxford (2022) An estimated 1.2 million people died in 2019 from antibiotic-resistant bacterial infections https://www.ox.ac.uk/news/2022-01-20-estimated-12-million-people-died-2019-antibiotic-resistant-bacterial-infections
【2】病死畜禽和病害畜禽产品无害化处理管理方法 http://www.gov.cn/zhengce/zhengceku/2022-05/13/content_5690117.htm
【3】 医院抗生素合理使用管理制度https://wenku.baidu.com/view/ecddbf0b7a563c1ec5da50e2524de518964bd336.html
【4】 Surface Immobilization of Human Arginase-1 with an Engineered Ice Nucleation Protein Display System in E. colihttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4968799
【5】利用冰晶核蛋白构建细菌细胞表面展示体系及其应用研究https://m.doc88.com/p-11847107229233.html