Team:GCGS China/Human Practices
Integrated human practice
Brief introduction
GCGS has attached great importance to the human practices section since the beginning of the project because we understand the close relationship between scientific research and society, industry , and the market, not to mention our desire to make complete products. When we conceived the human practices campaign, we were greatly inspired by the stakeholder segmentation of the 2020 IGEM TUDelft team, which divided the groups of stakeholders we wanted to engage into three categories:Science: Some of the authors of the literature we refer to, professors of oceanography and Aquaculture , etc,industry: farms, fishmongers, fish restaurants, food safety microbiome testing enterprises and civil society: Government food safety management agencies, relatives and friends, and ordinary people concerned about fish corruption
However, in the subsequent human practices campaign, we encountered situations where we were unable to get in touch or did not contribute to the project's perspective, so only the stakeholders remaining:
- Science: Professor of Oceanography and Aquaculture
- Industry: farms, food safety microbiological testing enterprises
- Civil society: Government food safety management agencies, relatives and friends, and ordinary people concerned about fish corruption
Brief summary
We did a lot of human practices activities, including interviews with national food safety management agencies' workers and offline aquatic markets public. We also designed questionnaires to help us investigate the background of our project. More importantly, throughout the whole process, we also did integrated human practice work to help us build and improve our project, for example the interview with doctor Zong facilitated us fixing the strip problem and finally we completed a whole functional test strip.
The interview with national food safety management agencies' workers
We are very sure that the problem of fish corruption exists, and timely detection can better protect human health. With some research done in the pre-stage of our experiments, we finally focused on a kind of bacteria that generally occurs when fish are corrupted——the Pseudomonas aeruginosa. After we determined that we were going to do a quick test of the Pseudomonas aeruginosa in fish, we first wondered if the contents of our project were unique, that is, whether existing fish corruption tests involved Pseudomonas aeruginosa. In terms of necessity, we must investigate whether there were relevant regulations on the breeding of fresh microorganisms in fish and the means to detect such corruption in our country's laws and systems. We found two workers in the relevant agencies of national food safety management (because their identity information is not allowed to show, so we only show written results), in the form of social media online text message communication, and telephone interviews, as well as to listen to their advice and guidance, reference to the integration of China's food safety law and other rules and regulations, to solve the following problems. See the detailed interview information at The interview with national food safety management agencies' workers
Our gains and feedback
Through the communication with the two interviewees and the inquiry and study of many food safety-related legal systems, we learned that:
First, the national quality inspection center sets up various testing standards for aquatic products to ensure that the food is harmless to the human body, not fresh or not. For example, there is no microbiological contamination testing link for primary aquatic products. Of course, according to our research, stale aquatic products do cause certain harm to the human body after consumption, so our projects and products must exist.
Second, the food safety traceability system is a regulation introduced in the past one or two years, which shows that my country's food safety control system is gradually updated, and it also shows the importance of my country's food safety management, which makes us think that it is very meaningful to study this direction.
Third, we understand that food processing plants do not need to conduct microbiological testing on the purchased raw materials and aquatic products and that the production process contains necessary sterilization processing steps. We have more clearly realized that the main consumer group of our testing equipment is the masses who cook independently. The promotion of the product also has a certain direction.
The questionnaire about public food safety awareness and customer expectation
After we determined the possible customer groups for future products, we decided to design a questionnaire to understand the habits of our potential customer groups regarding fish consumption. The questions we designed included personal information about the masses, the habits of cooking fish, and their opinions on eating fish, etc. We mainly wanted to understand the potential customer groups' attention to fish corruption and their expectations for the product. The following picture is the bref content of our qunestionaire. And the detailed information of our qunestionaire can can be seen at Questionaire 1
Fig 1. The analysis of our questionnaire 1
We were very satisfied with the results of the questionnaire survey, not only verified that many of our guesses were correct, such as fish cooks would judge the freshness of fish, but also willing to accept food safety-related science, and some data were more subversive, such as nearly 30% of people would not encounter a fish backlog, which meat that they often cook fresh fish, and later will not judge the freshness ratio (nearly 10%) or a certain gap. Even more surprising was that nearly 5 percent still ate fish when they judged it to be not fresh, and more than 10 percent still did not think it was necessary to accept more food safety-related knowledge. Of course, the questionnaire was limited, first of all about the sample size, our questionnaire was filled in less than 300 people, followed by GCGS members around relatives and friends and other groups, so the results and reliability of the questionnaire were also worth considering. But the results of this questionnaire at least indicated that there would be some demand for our products and many people would like to learn more about food safety, and also lay the foundation and theme for the upcoming public science content - fish food safety.
After we realised the demand for public education on food safety, we decided to do a series of educational activities . In addition, we utilized this opportunity to bring knowledge of synthetic biology to more people. Our education and communication process can be seen at Education and communication
Product design stage with the help of interview activities
Introduction
In the middle and late stages of the experiment, we started to design the product and a series of experiments on the product.
Our target product form is ALFA (aptamer lateral-flow assay) test paper. The most common related test strip on the market is the pregnancy test stick, which uses LFIA (lateral flow immunochromatography assay). Our product is ALFA based on aptamer, but unlike the sandwich method used in pregnancy test kits, we chose the competitive test principle because it was a more portable method and there were enough binding sites on the upper and lower surfaces.
Then we were focusing on other problems and factors related to the product designing work, for example, the customer survey. We also met some difficulties and solved them in human practices way. The content below is the results and gains we've made.
Interview with food testing company & questionnaire 2: about customers' needs and wants
When the project progressed to the product design stage, we decided to once again explore the public’s expectations and needs for fish spoilage quick inspection products in the form of a questionnaire. This time the questionnaire will be more specific and focused on the product. Since we were very worried that many questions in the previous questionnaire are duplicated or similar to the questions in this questionnaire, we also want to emphasize our focus. After designing the draft questionnaire, we asked the SMS team for their opinions. They gave us very reasonable suggestions, and our communication process will be shown in the collaboration session Collaborations. As for the result and the gains, most people who fill in the questionnaire have demand for our products and some suggestions for the use of the products. We have taken them into consideration in post-production and hardware design.
At the same time, we also really wanted to know the relevant information about such products in the market. We understood that there were no detectors directly sold to the market. Only some companies would provide food testing services. They would receive food samples from customers and use them in the laboratory. The inspection process was completed in the final delivery of the inspection report. Because we wanted to learn more about the relevant inspection service rules, we called Qingdao Chengyu Food testing laboratory Co., Ltd and learned some information about prices, inspection requirements, designated and special inspection services through a short telephone interview. In this interview, we learned about the testing services and quotations on the market. We also recognized the unique nature of our quick inspection products. We also learned about the necessity of testing from the business perspective of testing companies, as well as some market conditions. While completing the interviews and questionnaires, we have further refined our product design and hardware （See our Hardware). And here we combined company interviews and questionnaire results analysis and present the detail in written form Company interviews and questionnaire results analysis
Fig 2.One of the food quality detection company we interviwed
Offline Interviews in the aquatic product market
In the later stage, we conducted offline interviews in the aquatic product market. Based on the offline public interview experience of the SZ-SHD team at the collaboration team experience exchange meeting (see Collaborations), we re-written some of the questions in our questionnaire and conducted relatively simple random interviews with the public. The answers collected in random interviews and the corresponding analysis can be seen at Offline interviews
Offline surveys made us realize that the public, including some fishmongers, still have ambiguous attitudes towards quick inspection of aquatic products. We also found that river fresh was generally sold as live animals, while seafood is sold as chilled fresh. People would observe whether the gills of fish were bright red, but fishmongers generally thought that whether the color of the fish eyes was black was a more accurate way of judging. And for different types of fish, the methods and standards for judging freshness by naked eyes were also different. The seafood stall owner told us that yellow croaker, a kind of sea fish, cannot judge the freshness by looking at the color of the fish gills, mainly depending on the firmness of the fish scales. We learned the different ways of judging different fish with the naked eye. We have also realized that this may be the focus of our product promotion. Because bacteria, Pseudomonas aeruginosa, and whether they already exist, will not lie to the test paper.
Fig 3.The interview pictures at the market
The interview with doctor Zong: work out test paper color rendering problem
The principle of the designed aptamer-based strip method was based on the competitive reaction between the DNA probe 1 (test line) and the target C4-HSL molecule to combine with aptamers. If target C4-HSL molecule was in the detection solution, it would combine with aptamer–GNP probe, decreasing the aptamer–GNP that could hybridize to the DNA probe1 in the test line causing the red color intensity to become weaker. In other words, the more target C4-HSL molecule in the solution, the weaker intensity in the test line; Regardless of the presence of target C4-HSL molecule in the detection solution, the aptamer–GNP probe would definitely hybridize with DNA probe 2 in the control line, ensuring the validity of the detection. We have successfully verified that Probe 2 can successfully bind to the test paper.
However, the initial attempt to construct the strip was only successful at the control line(figure 4 left), the negative results should express color in both test line 1 and test line 2. Since the Great_bay SCIE 2020 made a similar hardware in last year's competition, we figured they may give us some advice. Then , it was an honor for us to have the chance to interview their advisor doctor Zong. We demonstrated our experiments in detail including the sequence and modification of our aptamer, test line and control line.
The test line sequence of the test strip probe was originally 5'-Biotin-CGGTCACTAGTACACCGCACCAGAAGCGGGCCCCG-3'. This was a complementary sequence corresponding to the C4-HSL aptamer. When the test paper was first made, the test line should also show color under a negative result, but the test line failed to show color. So we consulted with doc Zong, he suggested adding a C6 spacer between the biotin and the sequence to separate the modified group from the sequence and avoid affecting the function of the sequence, better able to unfold and combine with aptamer, so our modified sequence is 5'-Biotin-(CH2)6-CGGTCACTAGTACACCGCACCAGAAGCGGGCCCCG-3'. This allows us to finally successfully produce a test strip that shows color in both lines under negative conditions.The experiment results can be seen below(figure 4), and also can be seen in proof of concept
Fig 4.the test trip before the modificaiton(left) of test line and after(right) under negative condition