Team:OUC-China/Human Practices
Overview
Project Establishment
Our team has always been wanting to create a biosensor. Our two candidate projects were biosensors detecting pathogenic bacteria and urine sugar respectively and both include fascinating ideas. For the pathogenic bacteria detector, we designed it to report the concentration gradient of targeted bacteria by expressing different chromoproteins, green, yellow and red more specifically, that is the reason why we named it “traffic light”. As for the urine sugar detector, we proposed it to be combined with the idea of intelligent furniture. We planed to make the biosensor integrated in the closestool. Every time the the user urinates, the biosensor would automatically measure his urine sugar concentration and report when its is abnormal, enabling a healthier and easier lifestyle. We found both these two concepts intriguing, so we had a team voting, and the urine sugar detection project won.
Then we elaborated the design and presented it to our PI, professor Chenguang Liu. Our design was highly appreciated by professor Liu, while he provided us pertinent suggestions. He argued that urine blood had already became a routine test index , thus rapid detection methods must be well developed. He recommended us to reconsider whether our design can excel among all kinds of other mature detection methods. We seriously considered professor Liu’s advise, and started a new round of academic paper reading. Then we noticed that antibiotic has been proved to be able to be detected by biosensor, but the design is currently straightforward and conceptional which definitely needs redesign and modification for better performance and manipulation. The more backgrounds of antibiotic we explored, the more excited we became. One one hand, antibiotic contamination has been exaggerated quickly in this decades and surely needed to be monitored and controlled. One the other hand, as one of the most typical environmental new pollutants, it is still lacks of widely applicable detection methods, which calls for brand new approaches.
Despite feeling a little pity, we decided to abandon the idea of intelligent urine sugar detecting closestool. However, since a biosensor is generally composed by 3 segregated models, namely detect module, logic circuit and report module, we only changed the detect module from sugar detection to antibiotic detection but maintained the later two modules.
Listen to Public Opinion: Questionnaire
We designed a questionnaire and distributed on May 8th,2021. We totally received 362 responds, all the information have received the permission of respondents for publishing. The graphs below showed the origins of the respondents(divided by provinces) as well as their backgrounds(divided as whether have biology and medicine related educational or working background ).
Here are main results we can conclude from the questionnaire：
1.Chinese public generally have inadequate knowledge of antibiotic
2.People with biology or medical background know more about antibiotic related knowledge.
3.Public are generally unsatisfied with antibiotic education.
4.Public prefer new medias thantraditional medias for science popularization. School and community are also been expected to shoulder educational responsibility.
Consultation with Experts
Professor Ru Shaoguo is a doctoral supervisor, registered environmental impact assessment engineer, editorial board member of Research of Environmental Sciences and Ansian Journal of ecotoxicology, executive director of Shandong ecological society and director of Qingdao ecological society.
Professor Ru Shaoguo’s research Fields: biological screening methods and biomarkers of environmental endocrine disruptors; Endocrine disrupting mechanism and ecological restoration of environmental secretory disruptors; Ecological risk assessment and ecological restoration of persistent organic pollutants.
After listening to a brief introduction of our project, professor Ru showed intense interests on it. He provided us abundant of information and suggestion, promoting our projects in the following aspects.
He firstly recommended us to read the paper of professor Zhu Yongguan, who is a academician famous for studying antibiotic in China. He did a thorough study of the distribution of antibiotic in China in 2013. Thus, reading his work may help us have a overall perception of the contamination status of antibiotic in China.
Professor Ru secondly told us a crucial feature of the usage of antibiotic in China, that is the proportion of antibiotics used for human is larger than that in developed countries. In China, the proportion of antibiotics used for human is almost as much as used for animals. This information helped us firm the idea of developing a biosensor detecting the antibiotics which is usually produced for human to adapt the circumstance in China.
Professor Ru had an abundant of experience contacting with aquaculture farms. After knowing that the antibiotics we planed to detect are erythromycin, chlortetracycline and tetracycline, Professor Ru told us that erythromycin and chlortetracycline are mainly for medical use of human, but they still exist in aquaculture industry due to misuse and illegal use. This information expanded our biosensor’ application scenarios that it not only can be applied on detecting the antibiotic in sewage of pharmaceutical factory, it can also be applied on farm sewage monitoring, serving both farmers and government departments responsible for supervising of water quality.
Professor Ru confirmed our guessing that the possibly being polluted water may exist in the sewage treatment plant, aquaculture pond, poultry and aquafarm wastewater. More specifically, sewage treatment plant may more likely to gather the human used antibiotic, while farm related regions may contain more animal used antibiotic. This information instructed our integrated human practice, for having more specific target people to interview and target places to conduct filed research and sample collecting(See more detail on our consultation with experts, filed research and sample collecting page)
Besides, professor Ru’s previous study showed that the nature water in estuary of Qingdao have considerable antibiotic concentration. This information inspired us to collect some nature water sample. We luckily took the scientific research ship owned by Ocean University of China, Dongfanghong No.2, sailed into Bohai, a continental sea of China, which had been influenced by human activity intensively and collected some water sample(See more detail in our sample collecting page).
Professor Ru kindly introduced us to a local sewage treatment plant for filed research and sample collecting. He also recommended us to collect the water sample in both water inlet and outlet, the comparison of the results may reflect whether the current sewage treating procedure can reduce the concentration of antibiotic. Even though this information may not very useful for our project design, but could be a crucial information for the Nanjing_NFLS, the iGEM team we later built partnership for their project is to develop a device degrading antibiotic(See more detail in our Partnership page).
Professor Ru showed great interests on our project design. He told us that the current method of antibiotic methods are mainly based on chemistry, and all of them need pre-treatment such as extraction. Our aim of developing a whole cell biosensor detecting antibiotic in the original water sample attracted him a lot. Professor Ru’s praise and expectation made us realize the importance of realizing this concept, which promoted us to design a series of experiments testing the activity of our designed whole cell biosensor in all kinds of possibly contaminated water sample. This definitely perfected our proof of concept section(See more detail on our proof of concept page).
Professor Ru also provided us a brain-fresh idea of biosensor. Comparing to chemical based sensors, biosensor can not only realize quantitative analysis just like chemical based sensor, but may also reflect the harm contaminants have on organism which traditional sensors can not achieve. Then we realized that we have met a dilemma: the fluorescence intensity which we designed to reflect the antibiotic concentration may also reflect the lethality antibiotic have on our whole cell biosensor. Because high concentration of antibiotic on one hand can initiate more cells(our projects’ chassis is E.coli) expressing fluorescence gene, on the other hand repressing the growth and replication of cells. Thus, in order to avoid the later circumstance, we have to make sure our whole cell biosensor is successfully inserted the antibiotic resistant gene(See more detail on our proof of concept page).
Professor Ru mentioned to us that different target users may have different requirements on sensor. For example, scientists usually seek for accuracy, while public prefers rapid and visualized reporting form. We didn’t seriously considered our target clients at the beginning stage of our project, but after this conversation with professor Ru, we tried to rethink of our strength and modify our design to meet the specific needs of our target clients(See more detail on our Entrepreneurship page ).
Professor Ru critically pointed out that the key difficulty of biosensor is to transform the chemical based input signal into electronic output signal. Before consulting professor Ru, our project design was ended with different tensity of fluorescence. But after professor Ru explained to us that in order to match the requirements of quantitative analysis, reporting the results by electronic signal is a must. And it is also a good way competing test paper analysis, whose results are usually evaluated by naked eyes, thus more of qualitative than quantitative. Being inspired by professors Ru’s recommendation, we designed our hardware which is a portable box containing the whole cell biosensors. We also developed a mini apps to cooperate with the hardware by analyzing the fluorescence intensity of the photo taken through a whole on the hardware, and reporting the corresponding antibiotic concentration(See more detail on our Hardware page).
Professor Ru told us that China is witnessing a series of policy reducing the usage of antibiotic for both animal and human use. This triggered the idea of having a thorough study of antibiotic related law, hopping to enrich our background knowledge on social level, and possibly provide some modification suggestions to resolve the antibiotic pollution issue beyond synthetic biology(See more detail on our Study and Refinement of Law page).
We luckily interviewed the process Engineer of Haibohe Sewage Treatment Plant, who gave us detailed introduction of the plant. Haibohe Waste Water Treatment Plant The designed water quantity was 200000 tons per day, and actual water quantity was 170000 tons per day, serving 530000citizents who lives in 24km2. The plant follows the Standard for discharge of pollutants from urban sewage treatment plants. The standard includes strict discharge limits of COD(Chemical oxygen demand), ammonia nitrogen, ss(suspended solid), total phosphorus, total nitrogen and so on. But new pollutants such as antibiotic are not included. Engineer Liu believed that the responsibility of degrading antibiotic belongs to the units who produce it such as the drug factory. But if antibiotic contamination be become a common and serious problem one day, and letting the sewage treatment plant shoulder the degrading responsibility become the most effective way, then the government should set the discharge standards, provide the antibiotic detecting and degrading methods.
Mr.Xue firstly admitted that Haibohe Sewage Treatment plant aren’t required to treat antibiotic, thus having no demand of antibiotic detecting. We then asked his opinion on what scenarios he reckon our biosensor may possibly being applied to. Mr.Xue critically pointed out that there would be demands when there were standards. We realized that the sewage treatment plant was a vivid example. People there, from the leader to the engineer, didn’t reckon antibiotic detecting device match the needs of the plant because there was no requirements of degrading.
But sewage treatment plant did care about antibiotic, Mr. Xue told us that before entering anaerobic tank, which is a crucial part of sewage treatment, there might exist the necessity of detecting antibiotic. Because if the sewage with low biodegradability entered the anaerobic pond, the anaerobic bacteria’ activity would be depressed. The so called “low biodegradability sewage” is more commonly seen in pharmaceutical factory sewage than other kinds of sewage because it contains antibiotic that inclusively kill bacteria. If the biodegradability of sewage is too low, the plant might add an extra anaerobic pond to make sure there are enough anaerobic bacteria working. This could be a very suitable case for applying our ALLPASS whole cell biosensor, but Mr. Xue added that biodegradability is a general idea including far more other environmental indexes that influence the living conditions of bacteria, such as pH, salinity, not just antibiotic. This means that a single antibiotic sensor is not enough for deciding whether the sewage can enter the anaerobic pond or not, not to mention the concentration of antibiotic has already been attenuated when enter the municipal sewage treatment plant, which has little effect on bacteria basing on the plants’ former experience.
Mr.Xue told us that the up stream corporations and social organizations have to reach their own standards before discharging into municipal pipeline which leads to the local sewage treatment plants. For example, hospitals have to reach the Discharge standard of water pollutants for medical organization, and farms should meet the Discharge standard of pollutants for livestock and poultry breading. However, we found antibiotic was mentioned in neither of these two standard.
After the conversation with Mr.Xue and Ms.Liu, we deleted hospital and farm from our potential user list. We then focused on pharmaceutical factory. But we know that even though hospital and farm may not the unities who need our antibiotic sensor most at current time, but are still our potential users. Once the standards are published, the needs boom. Besides, if the organization who produces antibiotic have strong environmental protection awareness, a cheap, fast and user-friendly device would be a good choice contamination monitoring.
Talking of antibiotic detection, what firstly came to our mind if the detection of the antibiotic residuals in food. We then consulted the the local Food and Drug Administration officer for the procedure and standards of antibiotic detection. The officer told us that antibiotic concentration was an important index in food safety indeed. And different type of food would be tested of different kinds of antibiotic, for example beef and mutton would be tested for tetracycline, while chicken would be tested for chloramphenicol. We then asked about the detection procedure, and learned the important information that the government would entrust a third-party testing agency to test the concentration of antibiotics. This information instructed us to plan another interview with the third testing company for it is an important stakeholder(See more detail in our Third testing company page).
The officer also told us that antibiotics were not detected in domestic water (tap water), this information complement to what the pharmaceutical factory told us(See more detail on our Pharmaceutical factory page), and expanded our application scenario.
Thanks to Nanjing_NFLS, we reached Mr.Jiao and had an online interview for we live in different cities, Qingdao and Nanjing respectively.Mr. Jiao got his master degree of environmental engineering from Jilin University. From 2010 to 2021, he engaged in operation and maintenance management of sewage, waste gas and solid waste in the environmental protection station of Hengrui pharmaceutical branch company. At present, he is incharge of environmental protection management in Hengrui pharmaceutical biotechnology company.
The two main drugs the drug factory produces are Daptomycin and Caspofungin. The former is antibiotic based drug which targets gram positive bacteria while the later is non antibiotic drugs. That is to say, Hengrui drug factory do has the discourse power of discussing antibiotic contained waste water related issues, especially the antibiotics for human use.
Mr.Jiao told us that every drug factory has its own sewage water treatment system. Drug factories usually have to reach two criteria before emitting into municipal sewage pipeline. One is about some general index, such as chemical oxygen demand (COD), ammonia nitrogen (NH3-N) and so on. Another criteria is the indicators of acute toxicity, called Discharge standards of water pollutants for pharmaceutical industry Chemical synthesis products category, for Hengrui drug factory specifically. After reading the documents, we found that antibiotic was not included as one of the acute toxicity substances.
However, Mr.Jiao told us that the drug factories which mainly produce antibiotic drugs follow another discharge criteria, which is called Discharge standard of water pollutants for pharmaceutical industry Fermentation products category. In this category, the limited proportion of sewage containing antibiotic was regulated.
As required by the Environmental Protection Bureau, a third party is regularly entrusted to test the acute toxicity of sewage. Water Quality-Determination of the Acute Toxicity-Luminescent Bacteria Test and Zebrafish(Danio rerio) eggs method are the common used method. The mechanism is that the acute toxicity of water sample to be tested can influence the luminous intensity and survival rate of luminescent bacteria and zerbafish eggs respectively. But this two methods have to be done by third party testing companies. Thus, Mr.Jiao told us that sewage treatment plants of pharmaceutical factory are looking forward to rapid testing products to detect the toxicity of sewage, but not just antibiotics.
Mr.Jiao emphasized that there is no suitable means to judge the toxicity of sewage to microorganisms at present. Like what Mr.Xue in Haibohe sewage treatment plant had already told us, the sewage treatment process is mainly anaerobic process and aerobic process. The anaerobic microorganisms in the anaerobic process are mainly methanogens, which are particularly sensitive to toxic substances. If the antibiotic concentration is too high, the anaerobic process cannot be functional, and the operation efficiency is very poor. At present, there is no good rapid detection product in China.
Mr.Jiao told us a story that in the first half of the year, a detection instrument in Canada measured the content of active substance ATP in activated sludge(containing anaerobic bacteria) to indicate the toxic substances. More specifically, it is achieved by comparing the amount of ATP in 100ml activated sludge under normal conditions and the change after immersion in toxic sewage. Mr.Jiao was super excited about this products but didn’t buy it for the price is very expensive. As such, Mr.Jiao was very pleased to hear that we are developing a rapid testing products for antibiotic detection with generally low price. He praised our products very useful and meaningful. And he considered it to have great market potential. Because it is impossible for the sewage station of a private pharmaceutical factory to be equipped with a HPLC and hire technicians.
At the end of our interview with Mr.Jiao, he motivated our directions for improvement. He told us that Hengrui pharmaceutical factory used to collaborate with Zhejiang University who screened for high salt and high toxicity tolerant microorganisms , using the bacteria in sewage station of Hengrui pharmaceutical factory as the resource. Mr.Jiao recommended us to consider the attributes of the sewage we planed to test with our whole cell biosensor, such as high salinity and high acute toxicity. Combing with the intention of Zhejiang University research group, we may use a high salt and high toxicity tolerant bacteria as our chassis organism.
Another motivation is another possible application scenario: waterworks. Mr.Jiao told us that the duality of antibiotics: are the inhibition of activated sludge bacteria and the negative influence on the drinking water when flow to natural water. Drinking water containing antibiotics is a risk to human health, thus detecting the concentration of antibiotic in waterworks is meaningful.
SGS is an internationally recognized inspection, identification, testing and certification organization. It is a globally recognized benchmark of quality and integrity. More than 93000 employees operate more than 2600 branches and laboratories around the world. Its core services can be divided into the following four categories: inspection, testing, certification and identification.
SGS, formerly known as the French grain shipment inspection institute, was established in Rouen in 1878. In 1919, the company was registered in Geneva and named Société Générale de Surveillance.
SGS vision: Their goal is to become the most competitive and productive service organization in the world. They are constantly improving and perfecting our inspection, identification, testing and certification services, and always provide first-class services for local and global customers.
We were luckily welcomed by a head engineer of SGS, and visited the chemical laboratories in its Qingdao branch office. We got to know that there are strict standards even for different parts of animal, for example the skin and muscle. So the food to be tested have to be separated and crushed, and this procedure was called sample making. SGS had intellectualization control experiment system, their food weighting results would be sent to a database automatically, making us realize that the quantification and automation can make the testing more effective and trustworthy.
Most of the test items SGS conducted are basing on GB(national standard), thus the accuracy was important. There was a strict and a little bit complicated procedure of testing.
Firstly, sufficient extraction is required to avoid false negative and false positive. After adding extractant, oscillation, homogenizer and ultrasonic can all effectively extract the target. Secondly, the testing material have to be purified in order to remov non-specific extracted interferents. There were a lot of approaches such as extraction, adding purification powder and SPE column. The third step was concentration. The method include “nitrogen blowing” and rotary evaporation concentration. The last step of pre-treatment was determination, which means settling the volume, which was usually 1 ml. Then comes to the testing section. The principle of LC-MS is to separate the substance to be measured from the impurities first, and then use mass spectrometry. It is a very selective means according to the qualitative and quantitative of ion peaks. Test the standard, empty sample and sample in turn. The final concentration is obtained by multiplying the machine reading by the constant volume and dividing by the weight. The engineer in SGS told us, the better the specificity of the detection instrument, the less pre-treatment is required. We realized that one of our products’ advantage was there need no pre-treatment for it has high specificity. We also proved that our whole cell biosensor can work effectively in open sea water(See more detail on our proof of concept page).
There was an important procedure, that is to add interior label and external labels. The labels were usually made by changing the protium intodeuterium, which would not change the properties but distinguishable. The interior label was added at the very beginning link: the target extraction, while the external label was added in the very last link: testing. This way could detect the influence of interferents and correct sample loss. The sample loss could be really high in some cases, such as 80%. Thus, if there was no interior and external label, the real concentration could not be detected with accuracy. This method inspired us so much, that we deeply understand the importance of controlling.
The engineer in SGS told us that the current concentration of antibiotic in environment is on the level of µg/kg. Thus, if our product can’t reach this detection limit, our results would be all negative. We felt thankful for this advice.
We visited an aquaculture farm named Heshengyuan, which is located in Qingdao district. The boss of this private enterprise Sun Zhangde warmly welcomed us. Heshengyuan aquaculture farm mainly co-cultures sea cucumber seedlings and shrimp. The farm produced 178000 cubic meters of tail water a day. The antibiotic is used in the seedling pond only for it was uneconomic to apply antibiotic in culture pond with much more water. At present, the price of antibiotics was more about 100 to 200 yuan per kilogram. A Small and medium-sized farms like Heshengyuan would spent at least 100000 yuan a year on buying antibiotics. The antibiotic the farm used include neomycin sulfate, florfenicol and so on. The choice of antibiotic were made according to the regulations on the use of antibiotics in aquaculture. It is a common knowledge that the antibiotic legal using are the ones can be degraded quickly(usually 24 or 48h). Mr.Sun told us the list of antibiotic in the regulation has the trend of shrinking. The antibiotics which this farm used before such as Penicillin, oxytetracycline and furans are all banned now.
The local government made some other actions. From this year on, the Environmental Protection Bureau began to set the aquaculture wastewater discharge standard, namely 3ppm for legally used antibiotic. The Fisheries Bureau and Ocean Bureau had held a meeting to issue standards to the private aquaculture farms. The farms were informed to reform sewage treating methods, for example installing protein filter, and get ready for the spot check.
We asked the usage of antibiotic in detail. Mr.Sun told us that the injured sea cucumbers were treated with antibiotics when changing pools. The use of antibiotics on the farm has decreased by 50% in the past two years. These years, China as well as the world has witnessed a decreasing trend of antibiotics usage, and the national requirements are becoming more and more strict. Nowadays, there are many antibiotic substitutes, for example applying probiotic spores, making it dominant bacteria to inhibit pathogenic bacteria, antimicrobial peptides and chitosan can to improve immunity. However, even the cost of probiotics is lower than that of antibiotics, the effect was slow to see. Besides, probiotics mainly focus on prevention of diseases, while antibiotics mainly focus on treatment. Especially when there were burst of large scale disease, antibiotics were required immediately. In conclusion, antibiotic are now still irreplaceable in some circumstances. The vaccine against shrimp disease has made rapid progress in China in the past two years.
We then asked whether the farm are interested in biosensor for antibiotic detection. Mr.Sun expressed his interests, but added that he would only prefer it when the cost is not very high. After knowing more detail of our products’ design, he was surprised at that our product can detect specific antibiotic. Even though our biosensor can only detect 3 antibiotic currently, he suggested us to extend it into a platform wich can detect a series of antibiotic, exploring a bigger market. We realized that the request of farm is different from that of pharmaceutical factory. Pharmaceutical factory wants something has broad spectrum detection ability which can indicate the general toxicity of sewage water, while farm wants the product to be sensitive to a specific antibiotic only.
At the end of the interview, we mentioned that antibiotic was probably a sensitive topic for farm, for the customers concerned a lot. But surprisingly, Mr.Sun believed that antibiotic related problem was unavoidable facing. Strictly following the regulations giving his confidence facing the customers and government honestly.
Field research and Sample Collection
We went to a local domestic sewage treatment plant named Haibohe for field research and sewage water sample collection. We learned the sewage treatment procedure and the factory environment. We totally collected 3 bottles of water in inlet, outlet and anaerobic tank respectively. We would like to know the concentration differences in different links of the treating procedure(See more detail in our proof of concept page).
We visited a local aquaculture farm for we believe it is one of the potential users of our biosensor. We got to know the breeding environment and the current needs of aquaculture farm. We also collected the the water in inlet and the tail water respectively for we would like to know the influence of applied antibiotic on water quality(See more detail on our proof of concept page).
We visited an international testing company named SGS to understand the international acknowledged testing procedure. We luckily entered their chemical laboratory and have a deep conversation with a lead engineer. We also got to know the economical cost as well as time cost of antibiotic detection. Through this field research, we confirmed the advantages of fast testing, and were convinced there would be a large market share for the rapid testing products especially for new pollutants just like ours!
Team members in OUC-China took a scientific research ship named “Dong Fang Hong 2” sailed in Jiaozhou Bay. Jiaozhou Bay is located in the middle of the Yellow Sea, Shandong Province. About 120.10-120.37 degrees east longitude and 36.06-36.25 degrees north latitude. It is a semi enclosed Bay, approximately trumpet shaped. The exit faces east, covering an area of nearly 500 square kilometers. It is a large and excellent harbor in China, thus had been deeply influenced by human activity. We collected a bottle of sea water near a wharf. We would like to know how our biosensors would be affected in sea water condition(See more detail on our proof of concept page).
Meet Ups
2021 China iGEM Online Meetup was one of the very first meetings organized, which was hold on May 29th to 30th. On May, most of the teams had just started their project. Although most of the concepts hadn’t been proved by data yet, the ideas were already fascinating enough. Through this meeting, we got to know DUT, who’s project was to degrade microplastic. They were the first group of people we encountered who were dedicated on solving new pollutants environmental problem just like us. We then developed a long term collaboration with each other, and established a new pollutants alliance together after knowing more iGEM teams doing related work(See more detail on our collaboration page and Initiator of New Pollutants Alliance page).
We attend the 8th CCiC meet up on Augest 27th to 29th. Even though we couldn’t made it to attend the conference in person, we attended the whole meeting online, giving a presentation and have a lot of communication with other teams. Through the meeting, we got to know ZJUT who built a strong partnership with us since then(See more detail on our partnership page).
CRISPR Conference was organized by TJU_China on Sep.12.2021, and other 11 iGEM teams including us participated. collect the problems from each team, and discuss solutions together.Our team and Tongji_Software paired in this activity.
This is the part our team and Tongji_Software contributed to the Question & Answer chapter of CRISPR APPLICATION GUIDELINES. It collected our teams’ questions and the answers we provide to each other after researching and deep mutual discussion.
Initiator of New Pollutants Alliance
New pollutants refer to chemicals that are not included in routine environmental monitoring, but may enter the environment and cause known or potential negative ecological or health effects, and may become the object of future regulations. At this stage, the main new pollutants of international concern include: environmental endocrine disruptors (EDCs), persistent organic pollutants such as perfluorinated compounds, antibiotics and micro plastics.
Through our previous questionnaire survey, some new pollutants have attracted public attention (such as antibiotics), while some (such as environmental endocrine disruptors) may have relatively low "popularity". Generally, the public does not understand the harm and pollution status of new pollutants. In fact, not only the public, but also the environmental monitoring departments and sewage treatment institutions (new pollutants may enter the environment from water bodies) do not have much involvement in the detection and treatment of new pollutants because new pollutants are not included in conventional environmental monitoring. At present, the detection, harm and degradation of new pollutants mostly stay in research institutions. There are many teams dedicated to promoting and solving environmental pollution in the iGEM team. Compared with the pollutants that have been included in the routine detection, there is still a greater vacuum in the detection and degradation of new pollutants, and it also creates opportunities for the application of synthetic biology. In 2021, we established the new pollutants Alliance (new pollutants alliance), hoping to gather forces to explore solutions to new pollutants from the multi-dimensional aspects of technology, system and culture, and make a better world together.
Alliance Members
In short
In detail
A.OUC_China：Due to the large scale use of antibiotic on human and animals, the remaining antibiotic in natural water has posed a threat on ecosystem and human health. Our project designed a whole cell biosensor with high performance of detecting antibiotic(taking tetracycline and macrolides as representatives). In order to overcome the restrictions of traditional restrictions of WCBs, we choose a fluorescent activated RNA aptamer(3WJdB) as the output signal to increase response speed.Moreover, we applied NIMPLY logic gate (comprising CRISPRi[3] and strand replacement reaction) to improve signal-noise ratio and dynamic range.
DUT_China：Our work presents a simple strategy to improve PET plastic degradation in river, lake, soil and biosynthetic microbe factories. The first step is to degrade PET in biology method, then we can use the degradation chemical products to produce more useful things.
C.Nanjing_NFLS：The degradation of residual tetracycline antibiotics in the environment by antibiotics is a practical problem related to ecology and human health in recent years. There is an urgent need to develop efficient and convenient tetracycline (aureomycin) antibiotic degradation technology. We found that microbial fuel cells can degrade aureomycin through Fenton reaction by modifying the cathode. At the same time, in order to improve the degradation rate, we located the biofilm of battery anodethe for biofilm activity is closely related to the efficiency of microbial fuel cell. In the process of film formation, the process of bacterial communication through chemical molecules is called quorum sensing. Through over expression of quorum sensing system in electrogenic bacteria, we greatly improved the efficiency of fuel cell, so as to improve the degradation efficiency of aureomycin.
D.HiZJU：17- α- Ethyl estradiol, EE2 for short, is an artificial estrogen with strong estrogenic effect. It is also the main component of short acting contraceptives. The intake of EE2 will affect the endocrine system of fish and human body, and cause a series of diseases, such as feminization of male fish, hysteromyoma, precocious puberty of children, decline of sperm motility and so on. EE2 is mainly concentrated in domestic sewage and industrial sewage. Because there is no effective degradation method, the concentration of EE2 in water increases gradually. Hi ZJU team is committed to degrading and detecting environmental estrogen EE2 by synthetic biological means. We introduced amoA gene and Hao gene from European nitromonas into E. coli to make the latter obtain the ability to metabolize EE2. In addition, we developed a high sensitivity method for the detection of EE2 under natural conditions by yeast two hybrid technology and artificial intelligence assisted protein transformation. Green fluorescent protein was added to the plasmid expression sequence of yeast, so as to visualize the EE2 concentration in wastewater.
E.SJTU-BioX-Shanghai：In the traditional rapid detection technology based on monoclonal antibody, the sandwich method often requires the target to have more than two different antigen epitopes. The competitive method is often cumbersome and has limited sensitivity. Therefore, the types of targets that can be detected by rapid antibody detection have certain limitations. More importantly, if some molecules do not cause the immune response of model animals, the preparation of monoclonal antibodies cannot be completed. At the same time, although the antibody development technology has been relatively mature, the relatively complex experimental operation and the inherent cycle of the immune process make it difficult to greatly optimize the time, human and resource investment required for antibody development. The subsequent protein expression, purification, modification and storage also have many difficulties for the cost is high, and the production and preparation cycle is difficult to be further compressed.Aptamer based detection technology can avoid many of these limitations. Therefore, SJTU BIOX Shanghai team hopes to use aptazyme liquid phase automatic screening technology combined with colloidal gold flow measurement analysis technology to build a platform that can quickly and automatically develop test strips for various new targets. Compared with the traditional immunocolloidal gold test paper, the production cost and production speed of the test paper products developed by this platform are expected to be greatly optimized.SJTU BIOX Shanghai team expects that it will play a great role in emergency response, large-scale multi-target preliminary screening and investigation, etc.
New Pollutants Workshop
We hold a new pollutants alliance online meeting introducing each other and discussing the collaboration form. The first meeting had only 4 iGEM teams participated, for SJTU-BioX-Shanghai joined the New Pollutants Alliance later. We a consensus that besides cross collaboration, it would be a great achievement to hold a new pollutants workshop, inviting experts outside of iGEM, welcoming public to get involved. Since resolving new pollutants is a comprehensive problem, but all of our members were just trying to grope for solutions through synthetic biology. Thus, listening as well as having conversations with people of other fields must be beneficial.
We designed 4 modules for our workshop, which were synthetic biology, rapid test, ecotoxicology and law area. We hope the conversation can include both academic experts as well as experts from company. Thanks to SJTU-BioX-Shanghai, we invited Shanghai Ruixin high-tech private enterprise who produces rapid testing instruments, making it a key stakeholder of the issue the majority of our alliance members engaged in. As for the fields of academic experts, we expected it to include ecotoxicology to help us update the frontier progress of new pollutants and synthetic biology to match the approaches we chose. Professor Wang Baojun, invited by HiZJU, had just started his new career in Zhejinag University(he used to work at the University of Edinburgh), and one of his research directions perfectly fit our concept: develop biosensors by synthetic biology. Besides, he was an experts in gene components and circuit design, making him a qualified teacher providing suggestions for the designs of the alliance members. We felt really glad for we are one of the very first organizations inviting him to give academic report after he came back from University of Edinburgh, England. The professors we invited who studies ecotoxicology came from Ocean University of China and Nanjing University of Technology thanks to the invitation of our team and Nanjing_NFLS. One of the features of new pollutants is as “new”. They are the newcomers to the nature environment due to the development of industrialization. Most of them are not well studied in a lot of aspects, especially its toxicity. However, unless their toxicity is generally well studied can we promote related regulations and laws. Thus, we also invited Students in Ocean University of China law school to join the conversation.
We hope that not only our members, but also the public gain from the workshop. So we made the workshop open to public and recorded it to publish on social media after the permission of the participants. We also made mind maps of the main contents of the workshop, please check it in our Education page.
Agenda of New Pollutants Workshop
Honored guests of New Pollutants Workshop
A.Dr. Wang Baojun
Title: Professor of Zhejiang University, Director of Institute of synthetic biology, Hangzhou International Science and innovation center; Vice president of Institute of biological and molecular intelligent manufacturing
Research direction: Synthetic biology; Gene components and circuit design; Biosensor and biological manufacturing; Biological computing and intelligent diagnosis and treatment.
B.Shanghai Ruixin Co., Ltd.
Shanghai Ruixin Technology Instrument Co., Ltd. is a high-tech private enterprise committed to the development and production of food safety rapid detection instruments, water quality analysis instruments, food safety rapid detection reagents and related software. The products are sold to China's agricultural system, market supervision system, smart market, canteen, supermarket, environmental protection chemical industry and other units, and some products are exported to India, Taiwan, Malaysia and Thailand. Its products have passed ISO9000 quality system certification for 15 consecutive years. More than 500 products have been formed, including pesticide residue rapid tester, multifunctional food safety analyzer, veterinary drug residue detector, grain mycotoxin detector, water quality analyzer, food safety rapid detection box, food safety rapid detection reagent and agricultural products Internet of things traceability system. It has obtained more than 30 national patents and software copyrights.
C.Dr. Zhang Xiaona
Title: Associate professor of Ocean University of China
Research Direction: Endocrine disrupting effect of environmental pollutants on fish and its harm mechanism; Explore the environmental health effects and pathogenesis of new organic pollutants using zebrafish as a model organism; Study on hazard mechanism and health risk assessment of new organic pollutants on shellfish.
D.Dr. Yuan Qingbin
Title: Associate professor of Nanjing University of technology
Research Direction: Environmental behavior and control technology of resistant bacteria and resistance genes; Identification and control of biological macromolecules such as viruses and DNA in the environment; Environmental application of nanotechnology and environmental effects of nano materials; Pollution characteristics and eco-environmental effects of micro plastics.
E.Dr. Wang Jun
Title: Associate professor of Ocean University of China
Research Direction: Research on marine ecotoxicology and environmental remediation, including current situation investigation of marine pollutants, biological toxicity, ecological risk assessment, environmental remediation technology, etc.
F.Dr. He Yide
Title: Associate professor of Nanjing University of technology
Research Direction: Study on the fate and toxicology of new pollutants in the environment
G.Cai Shiyu, Guan Yuezhi, Tang Linrong
Shiyu Cai, Undergraduate in LLB of Law School, OUC & James E. Rogers College of Law, UA
Lezhi Guan, Undergraduate in LLB of Law School, OUC & James E. Rogers College of Law, UA
Linrong Tang, Undergraduate in LLB of Law School, OUC & James E. Rogers College of Law, UA
The feelings and rewards of joining New Pollutants Alliance
A.OUC-China:
As the initiator of the New Pollutants Alliance and the organizer of New Pollutants Workshop, we feel that we are the team harvest the most. We initially focused on antibiotic only. But through the meet ups we attended, we found there are other teams who are also dedicated to new pollutants problem resolving by synthetic biology. We then came up with the idea forming a New Pollutants Alliance. All of our alliance member chose developing new pollutants detecting or degrading approaches, we then face some similar difficulties like visualizing signal reports, improving degradation rate, lowering detection rate and so on. A close alliance can culture cross collaboration, for example we had collaboration with DUT, and developed partnership with Nanjing_NFLS(See more detail on our collaboration and partnership page). Since we got to know each other through meet ups and workshops, we then came up with the idea holding a workshop together. But this time, inviting people outside of iGEM. Because scientific research may be at the forefront of solving social problems such as new pollutant pollution, but it must require the joint efforts of all sectors of society. From laws, enterprise, to public awareness.
The workshop turned out to be very successful. The conversations with synthetic biology inspired the members’ project design, and the conversation with the experts in ecotoxicology enriched every team’s background. There were even further collaborations between our members and the invited experts. We received the email of the participating experts who thanked us for holding this workshop and looked forward to attend more. This is definitely one of the most pleased moments as the organizer of new pollutants workshop. The alliance is temporary, but there must be “latecomers” in the future for there must be more iGEM teams in the future trying to use synthetic biology approaches resolving new pollutants related problems.
B.DUT_China:
Honored to participate in the workshop of the New Pollutants Alliance composed of OUC-China, Nanjing_NFLS, HiZJU, DUT and SJTU-BioX-Shanghai and have a deep understanding of other iGEM team. Our team’s project is plastic degradation, which also includes the degradation of microplastics in new environmental pollutants. Our team’s ultimate goal is degrading environmental pollutants same as the one of OUC-China and other teams. In the meeting，we also learned about other pollutants besides microplastic——persistent organic pollutants, antibiotics, and the need for pollution control. New pollutants may enter the environment and cause known or potential negative ecological or health effects. These substances may become objects of regulatory management in the future. These compounds have attracted widespread attention from all walks of life, and they are also a hot spot in scientific research. We also learned about environmental related laws and testing standards at the meeting, which can help our projects go to the market better. Hope that our team can work together with other iGEM teams and contribute to the improvement of pollutant management and environmental problems.
C.Nanjing_NFLS:
We are pleased to be part of the New Pollutants Alliance. In previous Human Practices, our team has come to realize the same problems that new pollutants face in our current society: inadequate legislation, incomplete monitoring systems, and toxicology that needs to be studied. The New Pollutants Alliance is an opportunity for us to look at this issue from a holistic and common perspective. During the workshop, our team discussed with experts, iGEMers, and people interested in this issue from the perspectives of research, public policy, and detection systems. This has certainly inspired our own project and deepened our understanding of the issue of novel pollutants.
D.HiZJU-China:
New pollutants refer to pollutants caused by human activities, which have clearly existed but no relevant laws, regulations and standards have been stipulated or are imperfectly regulated, and endanger life and the ecological environment. The environmental estrogen targeted by HiZJU-China is one of them. During this year’s iGEM trip, we met teams that targeted the degradation of the other new pollutants. We formed a new pollutant alliance, held online meetings to share ideas, and jointly organized a workshop to optimize our projects. The workshop also played a role in disseminating science, which made the relatively unfamiliar concept of new pollutants no longer the elephant in the room, and can enter people's hearts and arouse their attention. To act together for the same goal, I think this is the meaning of the iGEM competition.
E.Shanghai SJTU-BioX:
Through the initiation of the OUC_China team, we participated in establishing the New Pollutant Alliance and conducted a series of discussions. Through the sharing lecture of other teams, we got a deeper understanding of new environmental pollutants, which greatly enriched our background and supported our idea of taking advantage of our platform to continue to develop new target detection kits.
In the process of participating in the workshop, we had detailed exchanges with teachers, company staff, and team members. Next, all of our projects are discussed, including the legislation issues on environmental pollutants. It is still a long way before the public realize the threatening of new environmental pollutants and regulating them formally, but it’s our duty to keep an eye on new substances. No matter detection or degradation, our goal is to come up with novel ideas and make the realization progress more quickly.
Finally, we jointly expressed our concern about the environment health and our expectation of making a difference to local environment.