Integrated Human Practice

General Human Practice

Theory only becomes a material force when it is linked to practice.

Overview

We believe that nothing exists in isolation from the world and is inevitably influenced by surrounding things. Only by immersing ourselves in society, becoming a part of it, learning from the whole society, understanding its needs, and being inspired by real-world issues and society, can we remember the original intention of synthetic biology to make the world a better place. We must leave the narrow laboratory and enter the vast new world.
We conducted an in-depth analysis of the multi-faceted impact of the project on human society and engaged in sincere and productive communication with a broad range of stakeholders. These stakeholders covered the entire industry chain, including vitamin producers, feed manufacturers and their buyers, beauty and cosmetics institutions and their customers, pet hospitals, aquariums and zoos, breeding farms, as well as medical personnel, patients, university professors, government officials, and students from various fields.
We innovatively established a new HP cycle framework—the TPNR framework (Theory - practice - new-theory - re-practice). This framework enables us to continuously reflect on and optimize our theories and methods, allowing us to consciously (Selbstbewusstsein) realize our original intentions.
We gained a deep understanding of societal needs and constantly adjusted the project direction in Practice, always moving toward our original intention. We organized a series of productive ethical seminars, debates, and joint blog activities and actively collaborated with other iGEM teams. This not only improved our own project but also provided valuable support to other teams, effectively promoting the development and progress of the entire iGEM community.
We firmly believe that our project not only represents the forefront of technological innovation but also embodies a profound understanding of and firm commitment to social responsibility. We have every reason to believe that our efforts will make the world a better place.

TPNR framework

At the very beginning, we realized that conducting human practices to the standard of a qualified iGEMer was a grand and complex task. In order to conduct effective human practices consciously (Selbstbewusstsein) rather than spontaneously, we sought methodology from dialectics and, based on it, created our own framework—the TPNR framework—to guide our human practices.
TPNR framework: Theory - practice - new-theory - re-practice.

Figure 1: TPNR framework

Dialectics believes that the foundation of development is the initial original intention, and is also its own result, which means the development of things is an ascending circle. Things travel a journey and return to the starting point, and need to constantly return to their initial original intention. Apart from the most abstract initial original intention, there are no fixed and immutable theories. Everything needs to be constantly sublated in the process of concretizing the abstract original intention. Any theory can be overturned as long as it is proven in practice not to conform to the original concept. Theories in practice move towards their opposite and are sublated, and sublated theories are replaced by new theories. The new theories will go towards their opposite and ultimately be sublimed in practice. All theories are necessary steps to realize the final initial original intention, thereby completing a complete circle.
Returning to iGEM, we should not subjectively determine the details of the project, nor should we let any theory and assumptions limit the direction of thinking. We must start from our original intention, which is to make the world better with synthetic biology, and establish corresponding theories. We should internalize the original intention of making the world better with synthetic biology in the entire iGEM process. Apart from the original intention and result of making the world better, any part that is proven in practice to be detrimental to this goal will be sublated. The practice of striving to make the world better will help us establish new theories, guiding the next practice of sublating the parts that do not fit the purpose and continuously cycling. All theories that help make the world better will ultimately converge to our original intention, which is to make the world better.

Figure 2: Human Practical Diagram

In human Practice, we will analyze and organize the suggestions and opinions obtained from society, critically examine existing theories, and integrate suggestions and opinions that will help make the world a better place into our system, while discarding parts that do not align with the facts, forming new theories—still committed to making the world better. We will also commit to reinvesting these new theories into new practices, analyzing how our iGEM project can make the world better from an upward perspective and reviewing whether the theories we have established are advantageous or disadvantageous in making the world better from a downward perspective. This cyclical framework is the fundamental guide for our conscious human practice.

Choosing β-Glucan

Before finalizing the topic, we underwent multiple iterations and found that the team members were very interested in the synthesis and efficacy of polysaccharides. After screening, we chose the biosynthesis of β-glucan. β-Glucan can enter the lymph fluid through the intestines, interact with immune cells, enhance their activity, and has anti-inflammatory, anti-tumor, antioxidant, moisturizing, and cell regeneration-promoting effects, and is also an important source of soluble dietary fiber. Therefore, it can replace antibiotics to solve the problem of abuse and can also be used in skincare products ^[1]. β-Glucan has been widely used in food additives, animal feed, adjuvant drug therapy, and skincare cosmetics. Influenced by the COVID-19 pandemic, global consumers' awareness of health and hygiene has increased significantly, and the β-glucan market has seen remarkable growth. Consumers are increasingly concerned about health, and demand has shifted from processed foods to nutritious, low-calorie, non-fatty, functional, and super foods. The economically efficient and environmentally friendly biosynthesis of β-glucan has broad application prospects. Its excellent properties and the superiority of synthetic biology methods guarantee our goal of making the world better. Choosing β-glucan is an important starting point for our project.^[2]

Figure 3: β-Glucan

At present, most methods for synthesizing β-glucans rely on chemical synthesis or extraction from grains and yeast, which have stringent conditions, numerous steps, low yields, and potential differences in structure and properties compared to natural β-glucans^[3]. Synthetic biology can solve these issues by efficiently producing through microorganisms under mild conditions, reducing costs and energy consumption; avoiding the use of organic solvents, reducing by-products, and being more environmentally friendly; producing products that are closer to natural structures with better bioactivity and safety; and being easy to scale up for industrial production using renewable resources, in line with sustainable development concepts^[4]. These characteristics make synthetic biology an ideal choice for β-glucan production, offering significant economic, environmental, and social benefits. Therefore, we are committed to achieving economical, efficient, and environmentally friendly synthesis of β-glucans through synthetic biology methods.

Determining the chassis organism

Human practices with Shandong Xinglian Pharmaceutical Co., Ltd. and professors from the Marine Life Institute.

Initially, we naturally chose Saccharomyces cerevisiae as the chassis organism, which showed that although we had established the TPNR framework, we were not adept at using it due to our lack of initial practice. In fact, throughout the iGEM journey, we made multiple mistakes, and behind every misstep was an unconscious failure to apply the TPNR framework.
After learning that Shandong Xinglian Pharmaceutical Co., Ltd. had considered producing β-glucan, we communicated with their production leaders to understand the existing β-glucan production methods and their shortcomings. The production leader explained that they had indeed considered producing β-glucan before, but they found that β-glucan produced by Saccharomyces cerevisiae was located inside the cells and required enzymatic hydrolysis and alkaline-acid extraction to obtain it. These methods were complicated, inefficient, costly, and produced a large amount of polluting waste. Eventually, they abandoned the idea of producing β-glucan for feed additive purposes. The production leader believed that if we wanted to use synthetic biology for large-scale β-glucan production, solving the complex and inefficient extraction methods of β-glucan was a necessary step.

Figure 4: Shandong Xinglian Pharmaceutical Co., Ltd.

The employees of Shandong Star Union Pharmaceutical Co., Ltd. have stated that the production of antibiotics has been strictly limited, and they are happy to see the emergence of antibiotic alternatives. Not only because they generally have the ability to produce β-glucan, if β-glucan is put into production, it can serve as a supplementary product for their already reduced antibiotics, but also because they see that the production of β-glucan as an antibiotic alternative can slow down or even reverse the development of bacterial resistance, which might lead to a relaxation of antibiotic production restrictions.

Figure 5: Production Manager of Shandong Xinglian Pharmaceutical Co., Ltd.

We believe that the pollution and high costs caused by complex and inefficient extraction methods completely go against our original intention of making the world a better place. To address the current difficulties in the extraction of β-glucan, we have communicated with two professors from the Marine Life Institute who have research in molecular biology. Professor Yang Xiuxia acknowledged the great difficulties in extracting β-glucan and recognized the possibility of our idea of adding tags to β-glucan to facilitate extraction. However, Professor Yang sharply pointed out that our idea to solve the problem was superficial and had not delved into the main reasons for the need for enzymatic hydrolysis and alkali-acid extraction in the extraction process—the process of lysing brewer's yeast to obtain intracellular β-glucan generates a large number of impurities.

Figure 6: Professor Yang Xiuxia

Mr. Lü Zhiyi also emphasized the difficulties in β-glucan extraction. He further explained that the high molecular weight and multi-hydroxyl group characteristics of β-glucan make it very sticky and prone to interact with other molecules. Therefore, during the extraction process, β-glucan is very easily contaminated with impurities such as proteins, lipids, and other polysaccharides leaked from the cells after their breakage. These contaminated impurities are often difficult to remove entirely through conventional extraction and purification steps.
Mr. Lü also pointed out that due to the stickiness and susceptibility to contamination of β-glucan itself, even with the use of high-precision separation techniques like high-performance liquid chromatography, there will still be problems with the low purity of the extracts and the difficulty of separating impurities. Additionally, Mr. Lü mentioned that although the commonly used enzymatic hydrolysis and alkali-acid methods can break down the cell walls and release β-glucan, these methods will also introduce new contaminants, further complicating the extraction and purification process. Therefore, Mr. Lü suggested seeking milder and more efficient extraction methods, or developing specific separation technologies, to reduce the impact of contaminants and fundamentally improve extraction efficiency and purity.
Mr. Lü Zhiyi's insights provided us with a valuable perspective on understanding the challenges of β-glucan extraction further. He not only gave a profound analysis of the bottlenecks of existing technologies but also pointed out the direction for future research and process improvements. By comprehensively considering the structural characteristics of the molecules and the optimization of extraction processes, we hope to find more environmentally friendly and efficient solutions to achieve the high-purity extraction of β-glucan, reduce environmental impact, and promote the goal of sustainable development.

Figure 7: Professor Lü Zhiyi

Both professors pointed out that we should not think about separating pure β-glucan after it gets contaminated but should look beyond the superficial problems in front of us and find the main contradiction of the problem—the contradiction between the massive pollutants generated by lysing brewer's yeast and our vision of efficiently obtaining pure β-glucan to make the world a better place. After communicating and practicing with the professors, we suddenly realized that according to the TPNR framework, we should abandon the uncritically accepted theory of using brewer's yeast as the chassis and communicate with more relevant personnel to update our theories. Given the actual situation of our university—Ocean University of China—we attempted to find more suitable chassis organisms from the ocean.
With the goal of finding chassis organisms from the ocean, we communicated with Professor Liu Guanglei, a professor of marine microbiology. Professor Liu pointed out that we should use a new type of marine-like yeast (Aureobasidium melanogenum BZ-11), which has excellent characteristics—this yeast can produce extracellular polysaccharides including β-glucan, avoiding complex extraction steps, and its production even exceeds that of brewer's yeast.

Figure 8: Professor Liu Guanglei

The naturally blind preset of choosing brewer's yeast as the chassis organism has proven the superiority of the TPNR framework and the necessity of adhering to the TPNR framework. We hope to learn, apply, and master the TPNR framework in the upcoming iGEM journey. Meanwhile, we also look forward to sharing the successful experiences of the TPNR framework with other teams, jointly promoting the development and progress of the iGEM community, and making the TPNR framework a part of iGEM.

Improvement Project

Collaboration with Shandong Helen Food Co., Ltd. and Human Practice in Beauty and Cosmetics Institutions.

Guided by the theory of marine yeast as a chassis organism, we quickly engaged in re-practice. In the process of culturing marine yeast and isolating and purifying high-quality β-glucan, we discovered the presence of pullulan. We then went to Shandong Helen Food Co., Ltd. and communicated with their leaders and laboratory staff to understand their current separation methods and apply them to our project. Through in-depth communication with Helen Food, we learned that pullulan is a very difficult and costly impurity to separate. This information significantly influenced our understanding and strategy adjustment for the project.

Figure 9: Shandong Helen Food Co., Ltd. Laboratory and Laboratory Staff

We learned that pullulan is a very difficult and costly impurity to separate. If we do not address the issues caused by marine yeast producing pullulan, it will be hard to achieve high purity and guaranteed quality of β-glucan, making its production economically and environmentally unfeasible. Helen Food experts shared their experiences and challenges in separating pullulan, making us realize that the existing separation methods may not be suitable for our production scale and cost control goals. This prompted us to abandon our old theories and begin exploring new theories that align with our original intentions, implementing the TPNR framework once again.

Figure 10: Laboratory and Testing Staff of Shandong Helen Food Co., Ltd. Laboratory and Laboratory Staff

After careful consideration, and due to the challenges in finding a more suitable chassis organism, we decided to try using homologous recombination to knock out the gene producing pullulan in marine yeast. This decision was based on our in-depth understanding of the project and inspired by Helen Food experts' recommendations. Through gene knockout, we hope to address the pullulan issue at the source, thus improving the purity and production efficiency of β-glucan. During this process, our project also had a positive impact on Helen Food, providing them with new insights and methods, giving them more options when dealing with the pullulan separation issue.
At the same time, our collaboration with Shandong Helen Food Co., Ltd. also inspired our experimental thinking. The lab introduced us to two main methods for determining carbon and nitrogen content in feed: the Weiding combustion method and the Kjeldahl nitrogen method. The Weiding combustion method measures carbon content by burning the sample, while the Kjeldahl nitrogen method measures the nitrogen content in the sample. Both methods are efficient and convenient, though they may not provide very precise content measurement, they meet the needs of industrial production and are widely used. These inspirations from social and professional fields helped us understand more feasible and reliable experimental methods in practical applications, making us realize that the high-precision content measurement methods learned in university may be inappropriate in factory settings and should be abandoned. Different scenarios have the most suitable ways to achieve original intentions. This view provided valuable technical references and methodological guidance for our future β-glucan concentration measurements, implicitly indicating the basic logic of our ethical considerations.

Figure 11: Shandong Helen Food Co., Ltd. leader

As one of the potential impacts of β-glucan, communication with beauty and cosmetics institution leaders and users revealed a previously overlooked issue: marine yeast secretes melanin to the outside, which, although harmless, leads to poor appearance of β-glucan, reducing its acceptance — people tend to prefer lighter or white β-glucan. Beauty users generally express that if our β-glucan could look more "beautiful," they would be more willing to choose it. Also, due to the challenge of finding more suitable chassis organisms, we decided to try knocking out the gene producing melanin in marine yeast to ultimately solve this problem.

Figure 12: Beauty and Cosmetics Institution Doctor

Figure 13: Beauty and Cosmetics Users

We communicated with Professor Chen Chang from the Chinese Academy of Sciences. Professor Chen suggested using nematodes to study oxidative stress. We might try feeding nematodes marine yeast and observing their redox state. Regarding cosmetics, the professor suggested exploring internal whitening products. Nowadays, many cosmetics only provide physical coverage, with ingredients hard to absorb by the skin. β-glucan is non-toxic and can be used as an ingredient for internal whitening products. Nematodes are a good model for studying antioxidation and beauty since they are transparent and easy to observe. If our product can reduce the accumulation of lipofuscin in nematodes, it could become an effective antioxidant and whitening product. These suggestions helped clarify our research direction and enhance the project's practical application value.

Figure 14: Professor Chen Chang and his students

Human Practices with Government Agencies

As a necessary step in implementing our project in the real world, we held in-depth discussions with several related government agencies, including the Animal Health Supervision Bureau and the Tengzhou City Animal Quarantine Declaration Point. During these discussions, we not only received valuable feedback and suggestions but also gained a deeper understanding of the practical applications of the project. The Nanfan Village government especially reminded us to pay attention to production safety and biosecurity issues, which are crucial for the smooth progress of our project. They believed that our initially designed kill switch to prevent the leakage of resistant-gene marine yeast was inappropriate because it involved toxic proteins, and the potential application scenario of β-glucan we envisioned was for human and livestock consumption, with possible contamination by toxic proteins posing a potential risk.
The Nanfan Village government also emphasized that if we want to use β-glucan as an alternative to antibiotics, there are many aspects that require further study. For example, the possible interactions between β-glucan and other drugs, and the long-term effects of β-glucan on livestock and human health, these all need to be verified through long-term experiments. These opinions made us realize that although β-glucan has significant antibacterial potential, its safety and efficacy must undergo rigorous scientific validation. We need to design and conduct a series of long-term experiments to evaluate its performance under different conditions and its interactions with other commonly used drugs.

Figure 15: Nanfanjia Village Government

Figure 16: Staff of Nanfanjia Village Government

The staff at the Tengzhou City Animal Quarantine Declaration Point also introduced us to a potential user group we had not previously considered—feed producers and farmers who need feed. It is now prohibited for feed producers to add antibiotics directly during the feed production stage, and β-glucan might be added to feed as a substitute for antibiotics. This suggestion opens up new application fields for our product. We learned that feed producers and farmers have an urgent need for antibiotic substitutes, and our β-glucan can meet this need. By further communicating with these potential user groups, we can understand their specific needs and usage scenarios, thereby optimizing our product design and application strategies.

Figure 17: Tengzhou City Animal Quarantine Declaration Point

Figure 18: Staff of Tengzhou City Animal Quarantine Declaration Point

In fact, it was the suggestions of these government agency staff that ultimately led us to the agricultural track. We realized that the demand for antibiotic substitutes in agriculture is huge, and our project has broad application prospects. By cooperating with government agencies, we can not only ensure the safety and compliance of the project but also use their resources and network to promote the practical application and promotion of the project.
In addition, our project has also attracted the attention and support of the Gaoxing Town China Animal Health Supervision Bureau. The staff of the Gaoxing Town China Animal Health Supervision Bureau showed great interest in our research results and were willing to provide assistance in future promotion and application. This not only provided us with valuable feedback and suggestions but also opened up more application scenarios and market opportunities for our project. Through collaboration with these social organizations, we further publicized the application effects of β-glucan and collected feedback from society. This feedback not only helped us optimize product design but also provided us with more market demand information, enabling us to better meet user needs. Our project was not only influenced by other organizations and individuals in society, but also had a positive impact on them through our efforts. By cooperating with various government agencies, we not only promoted the application and promotion of β-glucan but also contributed to the health and safety of the entire society.

Figure 19: Gaoxing Town China Animal Health Supervision Bureau

As another potential impact area of β-glucan, we had in-depth communication with the person in charge of Shandong Chengji Feed Co., Ltd. They indicated that in the first decade of the 21st century, antibiotics were still allowed to be added directly to feed. However, after the start of the second decade, the policies regarding antibiotics have been tightening continuously. The Chinese government now prohibits the direct addition of antibiotics during feed production. This policy change reflects the global increasing concern over the abuse of antibiotics, especially in the feed and farming industries, where the overuse of antibiotics has led to the development of resistant strains, posing a serious threat to public health.
Feed producers now generally add acidifiers, antimicrobial peptides, tannic acid, probiotics, essential oils, bacteriophages, and other substances to feed as supplements. However, feed producers stated that the effects of these additives are generally questioned, and to a large extent, farmers still add more effective and direct antibiotics themselves after purchasing the feed. This information aligns with what we obtained from the Tengzhou City Animal Quarantine Declaration Point. These substitutes, though somewhat effective in antimicrobial action, are often not as direct and obvious as antibiotics, leading farmers to still prefer using antibiotics in practice, which contradicts the original intention of the policy.
Through communication, we learned that although the unit price is relatively high, the amount added is much lower than bulk raw materials, so feed producers often do not mind a slight cost increase caused by adding a small amount of high-value substances like β-glucan to the feed. However, feed buyers are generally very price-sensitive, and price is often the key factor affecting their purchasing decisions. This reveals a contradiction in the market: producers are willing to pay higher costs for efficient and safe additives, but the final buyers, namely farmers, are highly sensitive to price, making it challenging to market high-value additives.
In response to this market demand, we have increased research efforts to enhance the production of β-glucan while reducing production costs. We not only explored the impact of various component contents in the culture medium on yield and production costs in actual fermentation processes but also used computer models to simulate optimal culture conditions and designed new types of fermenters to achieve the greatest social benefit at the lowest cost. We believe that through these efforts, β-glucan can not only become an effective substitute for antibiotics but also be attractive in terms of price, thereby promoting its widespread application in the feed industry.

Figure 20: Shandong Chengji Feed Co., Ltd.

Professor Xu Bin's human practice

We are committed to maximizing β-glucan yield through high-density cultivation of pseudo-oceanic yeast. However, we discovered that the synthesis process of β-glucan in pseudo-oceanic yeast consumes ATP and relies on oxygen. The weak oxygen uptake ability of these yeasts limits the yield of β-glucan. We tried solving this issue by aerating the culture medium, but practice showed that this theory did not completely resolve the problem. This is due to the inherently weak oxygen uptake ability of pseudo-oceanic yeast. After discussing with Professor Xu Bin, we decided to introduce a new oxygen uptake system to address this problem. This not only provides pseudo-oceanic yeast with a means to obtain oxygen but also opens up vast opportunities for making the world a better place.

Figure 21: Professor Xu Bin

Human practice with Shandong Chengji Feed Co., Ltd. and Rizhao Marine Fisheries Resource Enhancement Co., Ltd.

The production manager of Shandong Chengji Feed Co., Ltd. mentioned that bulk materials like soybeans and corn have already been automated for feeding, but small or dusty materials still require manual input to ensure accurate dosing. As a small-quantity additive, β-glucan might still need manual feeding by feed manufacturers, increasing labor costs. This insight provided us with valuable understanding of the current limitations in handling small quantities on feed production lines, inspiring us to improve feeder technology for precise dosing of small quantities to reduce labor costs.
The production manager also mentioned that feed requires high-temperature treatment at 95°C before forming to gelatinize starch, enhance nutrition and taste, and sterilize. A new concern was raised about whether β-glucan can retain its sterilization effect after high-temperature treatment. This prompted us to investigate β-glucan's stability under high temperatures. We reviewed literature and conducted experiments, proving that β-glucan can maintain structural stability and functionality at temperatures exceeding 120°C.
Our collaboration with Shandong Chengji Feed Co., Ltd. not only helped us understand the practical needs and challenges of feed production but also made us realize how technological improvements can influence and optimize the production process. By improving feeder technology, we can lower labor costs and enhance production efficiency, offering greater economic benefits to feed manufacturers. This technological improvement supports our project and provides new solutions for the entire feed industry.
Our research and experiments also provided scientific evidence for feed producers, alleviating concerns about the effectiveness of β-glucan after high-temperature treatment. Through our efforts, feed producers can use β-glucan more confidently, reducing reliance on antibiotics and promoting safer and healthier feed industry practices.

Figure 22: Shandong Chengji Feed Co., Ltd. feed production line

We communicated with Rizhao Marine Fisheries Resource Enhancement Co., Ltd. The head mentioned that aquaculture is mainly divided into two stages: factory farming on land and cage farming in natural waters. In factory farming, fish are placed in large pits and fed manually; in seawater farming, machines feed the fish. Microbial environment management and chemical methods like hydrogen peroxide are used in aquaculture to ensure fish health, with complex harmless treatment required before water discharge to avoid impacting the marine ecosystem. In contrast, β-glucan treatment is simpler, requiring only sedimentation and composting with fish waste, ensuring we remain on a path to making the world better and taking up corresponding responsibilities.

Figure 23: Rizhao Marine Fisheries Resource Enhancement Co., Ltd

We also learned that precise feeding is essential for livestock, reducing feed waste and costs. To meet the need for precise feeding and waste reduction, the head introduced a method where small amounts of feed are given first to attract the fish, followed by more feed once they gather, ensuring it is eaten before sinking. They also prefer low-density feed, which extends floating time and reduces waste. The head also said that experience can determine the appropriate feeding amount to ensure all feed is consumed, avoiding waste. Besides reducing waste, precise feeding avoids water eutrophication pollution, benefiting shrimp health by preventing diseases caused by overfeeding. We also plan to design a machine that floats in the sea for remote feeding and control, reducing labor costs and achieving precise feeding as part of our hardware.

Figure 24: Rizhao Marine Fisheries Resource Enhancement Co., Ltd.

Human practice with seahorses and Shandong Helen Foods Co., Ltd.

Besides common fish farming, we also communicated with breeders of a special species—seahorses. Unlike livestock and typical fish breeding with ready-made feed, seahorse breeding relies on frozen and freeze-dried fish and shrimp for feed. This method is prone to spoilage and diseases due to poor sterilization compared to high-temperature treatment. Thus, seahorse breeders must focus on food preservation and disease prevention. Seahorse breeders also introduced pet feed producers as major consumers of freeze-dried feed, believing that β-glucan in freeze-dried feed can reduce residual bacteria and lower disease risk.

Figure 25: Seahorse breeding farm

Freeze-dried feed is often used to preserve nutrients and is largely used for pets. We returned to Shandong Helen Foods Co., Ltd. and conducted new interviews with pet hospital doctors. The head of Shandong Helen Foods Co., Ltd. confirmed that pet feed often undergoes freeze-drying, but this method has poorer sterilization than factory high-temperature treated feed, possibly causing pet and seahorse diseases. β-glucan mixed with freeze-dried feed can reduce diseases, an unexpected application scenario. However, the head noted that pet feed markets often prioritize additive-free products as a selling point due to pet owners' concerns about additives. Unlike well-regulated human food additives, pet feed additives lack sufficient research and legal restrictions, leading to pet owners' preference for additive-free feed. The head advised seeking support from pet hospitals and professional zoos or aquariums to prove β-glucan's safety and immune-boosting effects for pets.

Figure 26: Shandong Helen Foods Co., Ltd. freeze-drying workshop

Additionally, Shandong Helen Foods Co., Ltd. staff mentioned an industry concern: livestock like pigs and sheep have sensitive smell and unpredictable food preferences; adding β-glucan to their feed might reduce their appetite. This insight provided valuable industry perspectives, highlighting the need to consider potential issues during product development. To address this, we plan to conduct experiments simulating actual feed environments to observe the appetite effects of different β-glucan concentrations on livestock. If confirmed, we intend to solve the issue by adding suitable attractants, validated scientifically to ensure safety and effectiveness. We will also work closely with regulatory bodies to ensure attractant levels are permitted, clearly labeling them on product packaging to inform buyers.

Figure 27: Freeze-dried pet feed

Through these measures, we aim to guarantee product effectiveness, meet market needs, and ensure safety and reliability. Our practice at Shandong Helen Foods Co., Ltd. helped identify and solve potential issues, providing new research directions and solutions. Our experimental results and technical improvements benefit our project and offer valuable references for additive development and application. This practice experience using the TPRN framework showed us that assumptions about β-glucan's neutral effects on livestock were wrong. Future practices must consider β-glucan's potential negative impacts on the appetite of the feed animals.

Figure 28: Pet feed shark treated with freeze-drying technology

The human practice at the affiliated animal hospital of Henan University of Animal Husbandry and Economy, Zhongnong Pet Clinic, Tiger Beach Ocean Park, and Dalian Forest Zoo

Doctors at the affiliated animal hospital of Henan University of Animal Husbandry and Economy agreed with the head of Shandong Helen Food Co., Ltd., that pet owners are indeed resistant to additives in pet feed. Veterinary doctors suggested using β-glucan as a standalone nutritional supplement in the pet field, recommended by doctors or purchased separately when pet owners deem necessary.

Figure 29: The affiliated animal hospital of Henan University of Animal Husbandry and Economy

Figure 30: The doctor of affiliated animal hospital of Henan University of Animal Husbandry and Economy

At Zhongnong Pet Clinic, doctors showed us an existing nutritional supplement that can improve pet immunity, which not only proved the market demand and confirmed the significance of our work but also showed that many intelligent minds are working with us to find antibiotic alternatives. From an economic perspective, doctors suggested that besides learning from their experiences, we also need to ensure the ability to compete with existing products. We plan to conduct competitive product analysis to investigate whether β-glucan has advantages and further improve it based on deficiencies. Meanwhile, we found that products aimed at improving pet immunity are relatively scarce in the market, suggesting it is a relatively blank market. However, the usage conditions and regulations are not clear. My suggestion is that pet hospitals should increase investment and research in this area, and we carried out edu to help them better understand immune enhancers.

Figure 31: Zhongnong Pet Clinic Doctors

Figure 32: Pet Nutritional Supplements

We also visited Tiger Beach Ocean Park and Dalian Forest Zoo to understand the potential application of β-glucan in aquatic and terrestrial animals. At Tiger Beach Ocean Park, we discussed with aquatic animal experts the effects of β-glucan on the immune system of fish and other aquatic creatures. Experts pointed out that the primary goal of aquariums is to provide an environment that closely simulates the wild, including natural food, so they are very cautious about adding β-glucan to feed. Although β-glucan can enhance immunity in fish and other aquatic animals, the feeding amount needs to be strictly controlled to avoid negative impacts on water quality. Experts' feedback made us realize that the application of β-glucan in the aquatic environment requires more precise and cautious management. It also inspired us to work on ethics, understanding that different origins lead to different value judgments and guide people to make different choices.

Figure 33: Tiger Beach Ocean Park

At Dalian Forest Zoo, we discussed with veterinarians and keepers the application of β-glucan in terrestrial animal feed. Veterinarians and keepers initially expressed willingness to try β-glucan, but after mentioning the concerns of aquatic animal experts from Tiger Beach Ocean Park, they began to consider these potential issues. Veterinarians emphasized the importance of providing a diet closest to natural as well, thus being very cautious when considering new ingredients. Although β-glucan has potential benefits as an immune enhancer, they suggested more research and trials are needed before its introduction to ensure its safety and efficacy.

Figure 34: Dalian Forest Zoo

In addition to pets and zoos, the farming industry occupies a large part of mainstream feed consumption. We visited and investigated companies and institutions managing large herds of livestock, including Rizhao Aquatic Group Corporation, Changming Dairy Farm, Laojuntang Huifeng Agriculture and Animal Husbandry in Pingshui Town, Xingyang City, Liran Ecological Agriculture Co., Ltd., Daixian Animal Husbandry Development Center, and Xinzhou City Agricultural Bureau.

Human practice with Rizhao Aquatic Group Corporation and Professor He Yan.

The head of Rizhao Aquatic Group Corporation said they agreed with the information we obtained from Rizhao Marine Fishery Resource Enhancement Co., Ltd., and detailed the use of antibiotics in fish farming. The head explained that the use of antibiotics is rare in both factory and sea farming due to low density and strong disease resistance of the fish. However, antibiotics are used more frequently in freshwater farming due to higher density and poorer water flow compared to seawater. The head also mentioned the difficulty in treating antibiotics in water, which makes our project more relevant for promoting environmentally friendly and healthy marine aquaculture to make the world better. The head also explained that precise feeding with machines is already common in factory farming but difficult in sea farming. To prevent feed waste and water pollution, feeding must be done when the water flow is calm, requiring workers to judge the right time. For example, sea farming must be fed during the alternating tides, a period of less than half an hour, making such machine construction difficult. Based on this information, we decided to temporarily abandon the original machine design and plan a more feasible scheme.

Figure 35: Rizhao Aquatic Group Corporation

We talked with Professor He Yan to understand the impact of adding β-glucan to fish feed on fish and the water environment. Professor He Yan agreed with Rizhao Marine Fishery Resource Enhancement Co., Ltd.'s view that precise feeding is achieved mainly by controlling feeding quantity so that most feed is eaten by fish. Professor He Yan also mentioned that long-term feeding of fish with β-glucan to maintain high immunity might affect growth rates. He suggested feeding only during the fry stage when immunity is weak. To reduce environmental impact, Professor He Yan proposed changing the usage method, adding β-glucan during feed production instead of spraying on finished feed. Although this avoids altering the production line, β-glucan is prone to dissolving and polluting the environment. Adding it during feed production not only reduces dissolution but also slows feed sinking, giving fish more time to eat, as noted by Rizhao Marine Fishery Resource Enhancement Co., Ltd. head. We plan to experiment to see β-glucan's impact on the water environment. Professor He Yan recommended using carbon detection to subtract control group data to determine the β-glucan content in water, or liquid chromatography if dissolution is low. These suggestions are very helpful to our research project. Following Professor He Yan's advice, we plan to add different concentrations and treated β-glucan feeds to different water environments to observe environmental impacts. Meantime, we will use liquid chromatography to accurately measure β-glucan content, ensuring data accuracy and reliability. Through these experiments, we hope to find a more scientific, effective fish feed formula to bring us closer to a better world. We also decided to change β-glucan usage to mixing during production, instead of spraying on finished feed.

Figure 36: Professor He Yan

Human Practices in Animal Husbandry.

At Changming Dairy Farm, we learned that they report their antibiotic use plans and strictly follow governmental approvals. They use antibiotics under national and veterinary guidance, and the treatment effectiveness is evaluated by veterinarians. From the discussion with Changming Dairy Farm's manager, we learned that excessive antibiotic use in the past led to antibiotics in milk, resulting in substantial economic loss due to discarded milk. The manager also expressed concerns about β-glucan potentially remaining in milk. We decided to conduct a series of experiments to determine β-glucan's metabolism and excretion in cows to ensure it does not remain in agricultural products or negatively affect their quality. We are also considering regular sample testing to monitor changes in the composition of agricultural products, ensuring each batch meets safety standards.

Figure 37: Changming Dairy Farm

Figure 38: Cows raised in Changming Dairy farm

In Huifeng Agriculture and Animal Husbandry, they usually use antibiotics only when cattle are sick and one to two months before calving as per veterinary guidance. The dosage is decided by veterinarians but not further reviewed by Huifeng, possibly leading to antibiotic misuse due to insufficient checks and poor drug selection control. They use traditional Chinese medicine instead of antibiotics during normal times, and they believe vaccines are the most effective way to prevent or treat diseases. However, vaccines are expensive, forming a large part of the total farming costs. In discussions with Henan Agriculture and Animal Husbandry Technology Co., we explained the mechanism of β-glucan, similar to vaccines in boosting animals' immunity by directly activating the immune system. Huifeng's employees showed great interest in β-glucan, thinking it could significantly reduce farming costs and improve overall efficiency if it could offer similar immunity-enhancing effects at a lower price.

Figure 39: Xingyang City, Ping Shui Town, Ancestral Hall, Huifeng Agriculture and Animal Husbandry

In Henan Agriculture and Animal Husbandry Technology Co., we found they focus heavily on preventive healthcare measures in their daily cattle management, reducing disease occurrence and spread through regular health checks and nutritional adjustments. They use antibiotics when necessary, but mainly improve feeding conditions and feed quality to boost immunity. Vaccines are still considered the most effective diseases prevention or treatment method. We learned they face similar cost pressures from vaccine use. Most vaccines are voluntarily administered by farmers for disease prevention, with only a few mandated ones. Henan Agriculture and Animal Husbandry shows willingness to try cost-effective and efficient β-glucan to maintain herd health and ease economic burdens.
Through interactions with Huifeng and Henan Agriculture and Animal Husbandry, we understood their needs and challenges regarding antibiotic and vaccine use. We received valuable feedback and suggestions, helping us optimize β-glucan application schemes to fit real farming needs. The positive feedback and willingness to try new technologies from Huifeng and Henan inspire our research team, affirming our efforts can improve the world and intensifying our enthusiasm for β-glucan research, ensuring its effectiveness and economic viability in practical applications.
Through practice communication with Huifeng and Henan Agriculture and Animal Husbandry, we promoted β-glucan in farming, providing new solutions to reduce farming costs and improve economic benefits. This bilateral interaction and cooperation advance our project and bring new hopes and directions to the entire farming industry.

Figure 40: Henan Agriculture and Animal Husbandry Technology Co., Ltd.

At Loran Ecological Agriculture Co., egg-laying chicken farms in Shengou Administrative Village, Jiajinkou Town, Gongyi City, Henan Province, and Phoenix Parent Duck Farm, Zhucheng Foreign Trade Co., they explained that poultry is prone to intestinal microbial diseases and the high density of breeding can cause large-scale outbreaks, necessitating regular antibiotic injections. Learning that β-glucan can directly activate intestinal immune cells, Loran's employees were delighted as poultry must cease medication at least one month before market, a risky period as diseases in this last month could lead to total loss. They are willing to try β-glucan to reduce disease incidence, despite the increased cost.

Figure 41: Le Ran Ecological Agriculture Co., Ltd.

Figure 42: Employee of Le Ran Ecological Agriculture Co., Ltd.

Figure 43: Poultry raised at individual egg-laying chicken farms in Shengou Administrative Village

Phoenix Parent Duck Farm, Zhucheng Foreign Trade Co., emphasized the critical role of breeding ducks in farming, directly affecting the quantity and quality of offspring. High-quality breeding ducks enhance genetic traits, economic benefits, and industry sustainability. However, once ill, they cannot maintain health during breeding periods without antibiotics. Phoenix Parent Duck Farm's employees expressed willingness to try β-glucan to reduce risks and ensure duck health. Their confidence and expectations in this new measure aim to contribute to the company's development and the breeding duck industry's optimization. The practices at Phoenix Parent Duck Farm and Loran Ecological Agriculture boost our confidence in our progress towards achieving our goals.

Figure 44: Phoenix Parent Duck Farm, Zhucheng Foreign Trade Co., Ltd.

At Dai County Livestock Development Center and Xinzhou Agriculture Bureau, they acknowledged that compared to regulated farm drug use, individual farmers' antibiotic use regulations need improvement, even with village epidemic prevention personnel or veterinarians' help. These preventions and veterinarians are the frontline against antibiotic misuse, but individuals rarely store antibiotics; sometimes these professionals, influenced by habitual thinking and past experiences, may administer unnecessary antibiotics. This insight informs our EDU work in vast rural areas where individual farmers lack farms' diverse anti-epidemic measures, thorough training, and effective supervision. They rely on personal or local professionals' experience for antibiotic use, possibly leading to antibiotic overuse in rural individual farms. We aim to deeply engage rural areas, promoting the dangers of antibiotic abuse and the advantages of β-glucan as a substitute, improving the world through education.

Figure 45: Dai County Livestock Development Center

Figure 46: Xinzhou Agriculture Bureau

Weifang Livestock Slaughter Quality Standard Innovation Service Center agreed with Nanfanjia Village government's feedback. They reminded us of the need to strictly control and test β-glucan residue to ensure meat safety, a previously unconsidered aspect. This higher safety requirement drives us to provide superior, safe meat products. The Service Center introduced quality standards and safety regulations in the slaughter process and suggested using a spectrophotometer to ensure compliance. They helped us develop a plan to monitor β-glucan residue, ensuring product safety. Our exchange informed us about β-glucan's effects on livestock and meat quality, prompting product improvement based on feedback.

Figure 47: Weifang Livestock Slaughter Quality Standard Innovation Service Center

Figure 48: Weifang Livestock Slaughter Quality Standard Innovation Service Center Staff

Survey-based Social Research and Human Practices.

Following government officials, feed producers, and farm leaders' suggestions, we conducted a survey. We asked 211 people and found many are worried about the mycotoxins and β-glucan in our project. Many are concerned about mycotoxins contaminating β-glucan and worry about β-glucan's safety. This shows significant concern about new technology in food production. Many worry about risks from livestock eating β-glucan produced by GM technology, but many are open to eating meat from animals fed β-glucan from yeast. This indicates some acceptance of new technology, but safety concerns persist. Over half mind eating meat with antibiotics. Our survey reveals the greatest concern is mycotoxins in β-glucan produced by yeast. Without avoiding mycotoxins and mitigating β-glucan residue risks, we cannot achieve our goal of bettering the world. We will continue to refine project design to ensure β-glucan's safety and efficacy, reducing risks. We will also strengthen communication, explaining our technology and safety measures to alleviate concerns. This survey emphasizes that research needs both technological innovation and public safety and acceptance consideration. We will maintain close communication, sharing research progress and findings, and optimize our project to truly benefit society.

Figure 49: Questionnaire survey report chart 1

90.05% of respondents believe there are antibiotic residues in the meat they consume, while 9.95% believe there are none.

Figure 50: Questionnaire survey report chart 2

When purchasing meat, 74.88% of respondents consider antibiotic residues, while 25.12% do not. 74.88% of respondents think the overuse of antibiotics in Chinese livestock is a serious issue, 9.48% do not think so, and 15.64% are unsure. 54.03% of respondents mind eating meat containing antibiotics, while 45.97% do not. 88.63% of respondents are concerned about the potential risks of livestock consuming β-glucan produced using genetic modification technology, while 11.37% are not concerned.

Figure 51: Questionnaire survey report chart 3

60.66% of respondents are willing to eat meat where β-glucan produced by marine yeast is used as an antibiotic substitute, while 39.34% are not willing. 91.47% of respondents are worried about toxin protein contamination in β-glucan, while 8.53% are not. 91% of respondents are concerned about the safety of the mechanism of action of β-glucan, while 9% are not. 42.18% of respondents believe that using β-glucan instead of antibiotics can make the consumer healthier, while 57.82% do not believe it can.
At the same time, we held an ethics discussion meeting for students to widely gather opinions from students and promote iGEM and our project. We also found that many people were concerned and resistant to the principle of β-glucan replacing antibiotics due to its similarity to bacterial polysaccharides. Because the structure of β-glucan is the basis of its function, we decided to enhance the promotion of the structure and functional principles of β-glucan to alleviate social fears. We will also indicate the structure and functional principles of β-glucan on the packaging to ensure users' right to know, proving that we do not infringe on anyone's rights, comply with ethical requirements, and ensure that we do not deviate from our original intention of making the world a better place. In addition to these discussions on how to leverage our expertise in iGEM, we also held extensive discussions and debates on the philosophical level regarding technological advancement and the ethics of synthetic biology.

Figure 52: Debate

Guided by the practical advice from communicating with government officials and conducting public surveys, we completely abandoned the theory of using toxic proteins to prevent the leakage of marine yeast-like organisms. Under the guidance of the new theory—avoiding the use of toxic substances to prevent the leakage of marine yeast-like organisms—we plan to adopt new methods in marine yeast-like organisms instead of the traditional suicide switch expressing toxic proteins to prevent leakage and ensure biosafety. According to our plan, the modified marine yeast-like organisms have lost the PKS gene and Ags gene, making it difficult for them to survive in the natural environment. However, the antibiotic genes introduced along with the plasmid during the transformation of marine yeast-like organisms still pose a risk of gene drift. To prevent this, we ultimately decided to eliminate the resistance genes. Initially planning to use CRISPR technology for deletion, we learned from previous lessons and communicated with Professor Yang Likun in a timely manner according to the TPNR framework. He pointed out that applying CRISPR technology in eukaryotic organisms like marine yeast has a low success rate and recommended using Cre-loxP technology to knock out the resistance genes. We promptly updated our theory and used Cre-loxP technology to knock out the resistance genes.

Figure 53: Professor Yang Likun

Communication with clinicians, patients, and ethicists.

As another potential impact of β-glucan, we communicated with clinicians, patients, and ethicists to obtain opinions from the source of antibiotic production, the medical community, and patients' acceptance of using β-glucan to enhance immunity, ensuring that the use of β-glucan as a potential antibiotic substitute and health product is ethical.
Through interviews with doctors and patients from different hospitals, we found that doctors in tertiary hospitals have a strong awareness of reducing antibiotic use, but the misuse of antibiotics is still widespread in many small clinics in China's vast communities. This strengthened our confidence that we are on the right path, as antibiotic misuse still exists, and we can indeed contribute to improving antibiotic misuse and making the world a better place.

Figure 54: Small clinic doctor

We also discussed the ethical issues of genetically modified foods and the development and use of new drugs with chief physicians in tertiary hospitals. Chief physicians believe that genetically modified foods should be approached with caution and strictly regulated through approval processes and oversight. However, they are open to drugs possibly produced using genetic modification or other high-tech methods from an ethical standpoint. They believe that the intake of food and drugs differs significantly, and the scenarios for drug use are more urgent.
The hospital is recruiting volunteers for new drug clinical trials. We asked about the key processes for new drugs to transition from the laboratory to clinical use. In most cases, pregnant women are excluded from clinical trials. However, the lack of experimental data can lead to a lack of available drugs for pregnant women when they fall ill. Does this indicate a flaw in this arrangement? The chief physician expressed a different view. He pointed out that pregnant women are not without available drugs when they fall ill. Although many drugs cannot be used by pregnant women due to the lack of clinical trials, there are still many long-established and verified drugs that can be used by pregnant women.
The chief physician explained that these drugs were initially allowed to be used in pregnant women in emergency situations. At that time, the severe threat to the life safety of pregnant women by diseases made it the best choice to use some drugs that were safe and effective for ordinary people. This does not violate ethical principles because it is the best way to protect patient interests. After multiple uses in different locations and times, the medical community reached a consensus that these drugs can be used by pregnant women without violating ethics. This development process of drugs usable by pregnant women greatly inspired us.

Figure 55: A diagram illustrating versatility.

According to the TPNR framework, during communication with the chief physician, we found that the past theory—that pregnant women cannot receive any unverified drugs—does not match reality. Therefore, we need to develop new theories. We thought of the essence of dialectics—specific problems require specific analysis. Indeed, ethical norms should not be immutable or fixed frameworks. The original mission of ethical norms is to make the world a better place. In specific situations, there should be unique ethical standards that match specific circumstances. As long as these unique ethical standards are proven in practice to be another step towards a better world, we should not rigidly adhere to a single ethical norm, expecting to solve all problems with a universal ethical norm. Instead, we should actively explore new theories in practice. This is the specific embodiment of the TPNR framework and it inspired us. In fact, this interview was the starting point for us to begin constructing the TPNR synthetic biology ethics framework.

Figure 56: Tertiary hospital doctors

Professor Qu Zhe also answered philosophical questions. He believes that many products in the past were put into use without ethical review, such as alcohol and tobacco. These products have existed for a long time and were allowed to continue being used without strict restrictions. This situation has led to many products being widely accepted and used in society for a long time. However, Professor Qu pointed out that now we should be more cautious about these and new products. We need to consider the potential ethical issues and hazards they may bring and ensure that these products have higher safety standards when used in society. Now we need to take more preventive measures and strictly review the ethical and safety issues of these products to prevent adverse consequences. Professor Qu also cited the stories of thalidomide and the radium girls to further illustrate the importance of ethical review, emphasizing the severe consequences of the lack of ethical review and safety standards in history. We believe that iGEM should learn from historical lessons when facing new products and technologies, strictly conduct ethical and safety reviews to prevent similar tragedies from happening again, and ensure that the public's safety and well-being are fully protected.

Figure 57: Professor Qu Zhe

We sincerely thank the ocean for providing us with invaluable marine yeast-like resources. These resources are crucial to our research and development. To further understand and fully utilize these precious resources, we specially visited the marine mangrove reserve in Guangdong. There, we had in-depth and fruitful communication and exchanges with local forestry bureau staff. This exchange benefited us greatly, allowing us to fully understand and witness the richness, uniqueness, and enormous potential of marine microbial resources. The ocean, as a great gift of nature, contains endless valuable resources. These resources not only provide important support for our team but also lay a strong foundation for all teams participating in iGEM. More importantly, they open up vast prospects and deep foundations for the future development of all humanity. In our great endeavor to transform microorganisms and even the entire nature for the benefit of humanity, the abundant resources of the ocean give us strong confidence and strength to transform the world. We firmly believe that relying on these valuable resources from the ocean, coupled with our relentless efforts, wisdom, and courage, we will surely be able to promote technological progress and social development, jointly ushering in a better future. The development and utilization of marine resources will bring great opportunities and challenges to human society, and we stand at the forefront of this exciting era, ready to contribute our part to a better world.

Figure 58: Forestry bureau staff

University Practice

Our iGEM team actively engages in exchanges and collaborations with teams from Shandong University, Beihang University, Peking University, Wuhan University, Huazhong Agricultural University, Xi'an Jiaotong-Liverpool University, Jilin University, China Agricultural University, and Nanjing Tech University. We shared detailed information about our project, especially regarding β-glucan research and its potential applications in various fields. We also participated in numerous technical discussions, learning advanced Wiki development techniques, bacterial colonization, and antibiotic resistance research. We conducted in-depth ethical discussions with other teams, focusing on potential ethical risks, public ethical awareness, and ethical considerations in biotechnology research. The team also collaborated on modeling projects, sharing our modeling methods and insights, and drawing new inspiration and methods from other teams' solutions. We participated in the CCiC and the "Synthetic Future Central China Exchange Meeting," where we not only received valuable feedback but also realized the potential advantages of using yeast in our project and the general lack of ethical considerations among iGEM teams in China. Through these exchanges and collaborations, we not only shared our research results and experiences but also learned a lot from other teams' advanced technologies and innovative thinking, which helps to enhance our project's research level and vision.

Exchanging with the Shandong University iGEM team SDU-China, we learned their techniques in Wiki development and simplified genome design concepts, and shared project management and technical implementation experiences.

We had in-depth exchanges with the Shandong University iGEM team SDU-China. We introduced our teams and projects to each other and shared technical information. SDU-China demonstrated their team logo design and advanced Wiki development techniques. We learned a lot about Wiki development from them, which will be very helpful for our future projects. We also shared the details of our project and our previous competition experiences and lessons. We provided them with our experiences in project management and technical aspects, especially how to solve problems encountered during the competition. We hope these shares can help them complete their projects smoothly. During the exchange, we also discussed the difficulties, innovations, and ethical issues encountered in our respective projects. The SDU-China team made breakthroughs in establishing polyploid E. coli, and their innovative ideas greatly inspired us. We also began to think about how to achieve similar breakthroughs in marine yeast-like organisms. We hope to establish polyploid marine yeast-like organisms and simplify marine yeast-like organisms, just like SDU-China did with E. coli. We introduced them to methods to improve cultivation efficiency by simplifying metabolic processes, hoping to help their research. Polyploid marine yeast can improve metabolic efficiency, increase cultivation density, and enhance gene redundancy, achieving more efficient cultivation. Simplifying marine yeast methods can further improve cultivation efficiency. These improvements are very important for our project, as they can reduce production costs and final user fees, and help our team go global, contributing to solving global problems. This exchange allowed us to learn the advanced technology and innovative thinking of the SDU-China team, and they also gained new inspiration from our experiences and research.

Figure 59: Offline collaborration with SDU-China

We discussed with Professor Tian Miao the possibility of reducing the genome of marine yeast-like organisms to improve fermentation efficiency. Professor Tian Miao recommended the JCUI organization and the Syn3.0 project. He also explained the practical difficulties of reducing the genome of marine yeast-like organisms and suggested that we could first try knocking out genes with multiple copies and genes useless for synthesizing β-glucan. This inspired us greatly, and after knocking out the pullulan and melanin genes, the ability of marine yeast-like organisms to produce β-glucan indeed improved.

Figure 60: Professor Tian Miao

Communicating with students from Beihang University, we gained new ideas about the potential applications of β-glucan in the aerospace field.
When we exchanged with a sophomore student majoring in Electrical Engineering and Automation at Beihang University, we mainly adopted a relaxed and casual conversation format. After briefly introducing the iGEM competition and our team's project, we had extensive and in-depth discussions on the details and philosophical aspects of biology and electrical engineering. This exchange not only allowed us to understand the ideas of students from different majors but also provided new ideas for future iGEM promotion work.
The Beihang student mentioned that he had heard of the iGEM competition but ultimately gave up participating because he was unsure how his professional knowledge could play a role in the competition. This feedback made us realize that in future promotions, we need to emphasize more that the iGEM competition is a cross-disciplinary platform where students from various professional backgrounds can find opportunities to showcase their expertise. We should provide more specific examples like "Introducing resources from the iGEM Community" to demonstrate how different majors can participate and play a role in iGEM projects, encouraging more non-biology students to get involved.
During the discussion, we also explored the potential applications of β-glucan. The Beihang student proposed an interesting idea of using β-glucan as astronaut food to help maintain their immunity, which may be lowered due to weightlessness and other reasons. This innovative suggestion opened up a new application prospect for our research, allowing us to see the potential value of β-glucan in the aerospace field. This not only expanded our research direction but also made us realize that our work might positively impact the world in unexpected ways, emphasizing the importance of learning from broad human practices. This exchange made us deeply aware of the importance of cross-disciplinary cooperation and inspired us to continue exploring more possible applications of β-glucan. We look forward to attracting more students from different backgrounds to join iGEM in the future and contribute to creating a better world together.

Figure 61: Sophomore at Beihang University

Exchanging with the chief technician of the Cardiovascular Research Institute of Peking University, we learned about the formation mechanism of atherosclerosis and gained potential research directions on the impact of β-glucan on the immune system. We had an in-depth exchange with the chief technician of the Cardiovascular Research Institute of Peking University. This exchange mainly revolved around specific experimental operations. After we briefly introduced our research project, the chief technician patiently provided us with many valuable suggestions and tips on experimental operations. These guidelines are very helpful for improving our experimental accuracy and efficiency.
Subsequently, the chief technician, from his professional perspective, explained in detail the formation mechanism of atherosclerosis and the various factors affecting its progression. He particularly emphasized the key role of foam cells (a type of macrophage) in the development of atherosclerosis. This information is very helpful for our in-depth understanding of the disease mechanism.
One of our research focuses is exploring the impact of β-glucan on the immune system. The chief technician believes that enhancing human immunity through β-glucan may have a certain impact on the development of atherosclerosis. This viewpoint provides a potential new direction for improving the treatment of atherosclerosis and proves that our research is moving in the right direction. This exchange not only deepened our understanding of atherosclerosis but also provided new ideas and motivation for our research.

Figure 62: Chief technician of the Cardiovascular Research Institute of Peking University

We explored the possibility of adding β-glucan to human daily food, focusing on its health benefits, impact on taste, and the importance of ensuring consumers' right to know.

We explored the possibility of adding β-glucan to human daily food. Luckin Coffee, as a well-known coffee chain brand, has maintained market competitiveness through frequent product innovations. In today's society, people are increasingly paying attention to the addition of healthy ingredients, such as the widely popular oat milk in recent years. The person in charge of Luckin Coffee in Rizhao City stated that adding β-glucan to coffee beverages is a promising innovative approach. This could attract more health-conscious consumers to try this new form of coffee beverage. However, the person in charge also expressed some concerns, particularly about the potential impact of β-glucan on the taste of coffee. They worry that the addition of this ingredient may increase the viscosity of the coffee, thereby reducing the user experience. To solve this problem, the team believes that experiments can be conducted to find the best balance between maintaining taste and increasing health benefits.

Figure 63: Luckin Coffee

Xana Hotel sometimes provides fresh food for guests. However, these fresh foods may contain parasites. Xana Hotel believes that consuming β-glucan along with fresh food may meet consumers' health needs when displaying fresh food. However, ethical dilemmas still need to be vigilant and require further research and discussion. Both stores believe that ensuring consumers' right to know is essential, especially when introducing new ingredients and ensuring food safety. Transparent information transmission is particularly important to gain consumers' trust.

Figure 64: Xana Hotel

Figure 65: Fresh food provided by Xana Hotel

Exchanging with the Wuhan University iGEM team WHU-China, we learned their unique methods and innovative thinking in in vivo colonization research and jointly discussed ethical issues and public ethical awareness surveys.

We had in-depth exchanges with the Wuhan University iGEM team WHU-China and introduced our respective projects to each other. We were very interested in WHU-China's research on in vivo colonization of E. coli, which inspired us to think about the possibility of yeast colonization in livestock and even the human body. We shared our experiences and methods in ethical issue research and jointly discussed potential ethical risks and public awareness. To promote extensive discussions on biotechnology ethics, we plan to create podcasts and organize offline lectures to share research results and experiences and promote the healthy development of biotechnology.

Figure 66: Offline collaborration with WHU-China

Exchanging with the Huazhong Agricultural University iGEM team HZAU-China, we learned their research methods and techniques on the risk of antibiotic resistance caused by pesticides and shared our experiences in applying β-glucan in aquaculture.

HZAU-China, like us, is committed to the agricultural field, and our common goal is to address the challenges of modern agriculture. With population growth and climate change, agriculture needs to shift to a more sustainable and safer model. HZAU-China has in-depth research on the risk of antibiotic resistance caused by pesticides, and we learned advanced research methods from them. OUC-Haide focuses on using β-glucan to replace antibiotics, and we shared our techniques and experiences with β-glucan. We had in-depth exchanges on technical details, innovations, ethics, and experimental safety, learning from each other. We plan to maintain technical exchanges and cooperation, jointly improving project levels and contributing to building a better agricultural ecosystem. We plan to organize the "Synthetic Future Central China Exchange Meeting" to facilitate deeper communication after project development and jointly promote the development of the agricultural field.

Participated in the China iGEMer Community Conference (CCiC), sharing project and ethical improvement experiences with other teams and realizing the general lack of ethical considerations among iGEM teams in China.

The China iGEMer Community Conference (CCiC) is a national mobile event independently initiated by iGEM teams in China, providing the largest exchange platform for iGEMers in China. This conference aims to provide a resource-sharing platform to promote mutual learning and exchange between iGEM teams and young synthetic biology enthusiasts in China. We were fortunate to participate offline in the 11th CCiC conference hosted by Xi'an Jiaotong-Liverpool University. During the event, we presented our project and created a poster to introduce our team's project. Through project presentations, we not only provided an overview of the project but also discussed ethical improvements with many other teams. In this process, we learned many valuable insights from other teams' feedback, especially realizing that ethical considerations among iGEM teams in China might generally be insufficient. This discovery inspired our future work direction, making us more focused on integrating ethical considerations into our projects. At the same time, we also shared our experiences and methods in ethical improvement with other teams, helping them better understand and apply ethical principles. We emphasized the importance of ethics in biotechnology research and provided specific improvement suggestions, hoping these shares could help other teams better address ethical challenges in future research. During this conference, we gained valuable insights, realizing that ethical considerations among iGEM teams in China might generally be insufficient, which also inspired our future work direction. Through exchanges with multiple teams, we not only learned their innovative ideas in project implementation but also shared our experiences and methods in ethical improvement. These exchanges benefited both parties greatly, and we reached a consensus on future cooperation intentions. Through this conference, we not only improved our research capabilities and ethical awareness but also helped other teams pay more attention to and improve their ethical considerations. We believe that only by finding a balance between technological innovation and ethical considerations can we promote the healthy development of synthetic biology.

Figure 67: Poster of CCiC

Participated in the ethical white paper writing of the JLU-NBBMS and CJUH-JLU-China teams, learning their project experiences in using synthetic biology methods to produce β-glucan and their systematic thinking on industry ethical issues.

We participated in the ethical white paper writing independently hosted by the JLU-NBBMS and CJUH-JLU-China teams. Through participating in the white paper writing of JLU-NBBMS, we learned their project experiences in producing antibiotic substitutes using synthetic biology and their thinking on project goals, methods, and potential benefits. At the same time, CJUH-JLU-China's white paper discussed the ethical issues in the antibiotic substitute industry, including safety, environmental impact, and data privacy. We gained valuable experiences and references from their systematic thinking and solutions. During the writing process, we also shared our experiences and insights in ethics and social responsibility, emphasizing the need for close integration of technological innovation with ethics and social responsibility in scientific research. These white papers also discussed the application potential of synthetic biology in medical and agricultural fields. Through these discussions, we not only learned other teams' views on ethical issues but also shared our research results and future plans in synthetic biology. We hope these shares can promote public understanding, guide scientific development towards a healthier and more sustainable direction, and provide strong support for future policy-making. Through participating in these white papers' writing, we not only learned other teams' profound insights on ethical issues but also laid a good foundation for creating new ethical frameworks. We believe that this two-way exchange of knowledge and experience will help promote the development of the synthetic biology field and ensure its applications meet ethical and social responsibility requirements.

Collaborated on modeling with CAU-China, NJTECH-China, and SDU-China, sharing respective modeling methods and ideas, and learning new methods and techniques from other teams' experiences.

We collaborated on modeling with CAU-China, NJTECH-China, and SDU-China and had in-depth discussions. We shared our respective modeling methods and ideas and explored the advantages and disadvantages and applicable scenarios of different models. Through exchanges, we learned from each other's experiences and techniques, improving the accuracy and effectiveness of the models. We unanimously agreed that modeling is not only a technical exchange but also an important part of team thinking collision. In the future, we will continue to collaborate, jointly promoting modeling innovation and laying the foundation for project development.

Figure 68: Modeling Discussion Meeting online

Participated in the "Synthetic Future Central China Exchange Meeting," receiving valuable suggestions from the judges and realizing the potential advantages of yeast in the project.

We shared our project progress with other teams at the "Synthetic Future Central China Exchange Meeting" and received valuable suggestions from the judges and participating teams. The judges suggested we re-examine the potential advantages of yeast, making us realize that although we initially chose other microorganisms, yeast still has many potentials in experiments and applications. We also shared our experiences in microorganism selection and application with other teams, hoping to help them make more scientific choices and adjustments. Through exchanges with other teams, we not only learned their innovative ideas and technical methods but also shared our practical experiences and research results, jointly promoting project optimization. We believe that this two-way exchange will help the project's in-depth development and ensure its success in practical applications.

Promotion

We confidently and proudly announce our achievements to the world. We are sowers, we are the publicity team, from public accounts to debate meetings, from team logo design to blog promotion, we report our work to society through various practices, promoting the iGEM competition and our project.

A concise and beautiful team logo reflects the combination of science and nature, incorporating Australian characteristics.

We designed a dedicated team logo and team uniform to enhance team cohesion and publicity effects. The design of the team logo and team uniform reflects our team spirit and culture. We put a lot of time and effort into ensuring that every detail perfectly showcases the team's philosophy. The main color tone of the team logo transitions from light blue to dark blue, echoing the school motto "Embrace all rivers, take them far." At the same time, it incorporates Australian characteristics, showcasing the distinctive feature of Haide Academy standing at the forefront and international exchanges. The simple double helix structure of DNA in the logo eventually transforms into the shape of mangroves, symbolizing the close combination of science and nature. Yeast and β-glucan are located on the same DNA structure, reflecting their relationship. The part of the DNA connected to the top of the yeast is designed in the shape of a bud, symbolizing new life and ecological cycles. The logo includes dot decorations, representing the texture of cells and the overall picture, giving the logo a dynamic feel and Australian flavor. The marine yeast expels melanin behind it.

Figure 69: Team logo

Choosing the college's public account for promotion improves promotion efficiency and effectiveness.

Initially, we planned to create our own WeChat public account to promote our goals and progress. However, after communicating with the Youth League Committee teachers and others with promotion experience, we found that creating and maintaining a new public account requires a lot of time and effort and may struggle to attract enough attention in the early stages. Therefore, we decided to use the more influential college public account instead. We believe that through the college public account, we can leverage its existing broad audience and resources to let more people know about our project and mission more quickly and effectively. At the same time, we can use the professional team of the college public account for promotion, making our information transmission more professional and comprehensive. This approach not only saves time and resources but also improves our promotion effect, allowing more people to know about our mission of creating a better world. This undoubtedly represents another leap under the guidance of the TPNR framework.

Organizing student gatherings to promote iGEM and inspire student participation enthusiasm.

We also organized a larger-scale student gathering to promote the entire iGEM, including our project. In addition to the aforementioned student from the Electrical Engineering and Automation major at Beihang University, this gathering included students from the Data Science and Big Data Technology major at the University of Jinan, Software Engineering major at South China University of Technology, Automation major at Shandong University of Technology, Clinical Pharmacy major at Shandong Second Medical University, and Electronic Science and Technology major at Harbin Institute of Technology (Weihai). These students' majors include engineering, data science, and medical-related majors, all closely related to iGEM and able to find opportunities to showcase their expertise in iGEM projects. During the gathering, after briefly introducing the iGEM competition and our team's project, discussion groups had extensive and in-depth exchanges on the ethics of synthetic biology. The participating students enthusiastically discussed their respective professional strengths and how to apply them in iGEM projects. For example, students from the Data Science and Big Data Technology major showed great interest in how to use data analysis to optimize experimental processes; students from the Software Engineering major proposed the idea of developing intelligent management systems to improve experimental efficiency; students from the Clinical Pharmacy major explored the potential of β-glucan in new drug development and clinical applications. The participating students firmly believed that they could showcase their professional strengths in iGEM and hoped to find like-minded friends in their respective schools to participate in the next iGEM competition together. Through this gathering, we not only gained many valuable suggestions and ideas but also strengthened our confidence and determination to continue promoting iGEM.

Figure 70: Students from various universities

Collaborating with other universities to create an online podcast to promote the ecological value of mangroves and call for their protection.

We also collaborated with many other universities to create an online podcast specifically to introduce the characteristics of mangroves and their ecological value. In this podcast, we not only introduced the various ecological functions of mangroves but also emphasized the enormous potential value contained within the mangrove ecosystem. For example, the marine yeast we utilize in our research project is a very good starting point for research. Through this podcast, we hope to let more people understand the unique aspects of mangroves and call on everyone to pay attention to and protect this precious ecological resource. Protecting mangroves is crucial for the overall health of the environment and the maintenance of biodiversity. Therefore, we hope to attract more people to join the action of protecting mangroves. We firmly believe that only through everyone's joint efforts can we achieve sustainable development of the ecological environment and leave a beautiful and rich natural heritage for future generations. We hope our podcast can become an effective educational tool, making more people aware of the importance of mangroves and taking practical actions to protect this key ecosystem. We look forward to every listener being inspired and participating in the practice of protecting mangroves, contributing to the ecological health of the Earth together.

Figure 71: Podcast

TPNR Synthetic Biology Ethics

There is only concrete ethics, not abstract ethics.

Overview

Ethicists generally believe that ethics is the philosophical exploration of a good life, which aligns with our TPNR framework. We have conducted research on synthetic biology and our project from three levels: meta-ethics, normative ethics, and applied ethics. The TPNR comprehensive synthetic biology ethics framework is the result of meta-ethics research. "Ethical Considerations of Antibiotic Substitutes for Livestock" is the result of normative ethics research. The specific analysis of the project is the result of applied ethics. The main goal of TPNR synthetic biology ethics research is to analyze and avoid ethical risks throughout the process of technology development, application, and governance from the perspective of people.
To implement the TPNR framework, establish synthetic biology ethics, and better represent the greatest social interest, we have attached great importance to ethical responsibility since the beginning of the project. We believe that only through responsible research, cautious application, and effective governance can synthetic biology truly benefit humanity and realize its potential. We conducted an in-depth analysis of the project from an ethical perspective and effectively communicated with stakeholders to ensure that the project ethically aligns with social values. At the 11th China iGEMer Conference, we actively interacted with other teams, understood project details, and provided ethical advice. Based on this interaction, we pioneered a new TPNR comprehensive synthetic biology ethics framework. This ethical framework not only helps us maintain the project's goals and original intentions but also provides a reference for other iGEM competition teams to improve their ethical shortcomings. We highly value the ethical impact of the project, actively contacting ethics experts, and organizing a series of activities including ethical seminars, debates, and joint blogs. Through these activities, we deepened our understanding of maintaining ethical boundaries in scientific research, increased public participation and transparency, and promoted public understanding and discussion of synthetic biology. We not only showcased technological innovation but also demonstrated a commitment to social responsibility. By adhering to TPNR synthetic biology ethics, we will ensure that the project is ethical, help other iGEM teams solve ethical issues, and contribute to creating a better world.

Reference

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