Team:UM Macau/Human Practices
Integrated Human Practices
Overview
Inspiration
How does HP help in this stage?
Biobricks Construction
How does HP help in this stage?
Functional Test
How does HP help in this stage?
Practical Application
How does HP help in this stage?
Background
Nanotechnology
Since the 1990s, the new wave of technology revolution driven by nanotechnology has brought us into a new age when scientists start to put their eyes on the property and potency of particles and molecules with ultrafine size. In fact, the word “nanotechnology” was firstly defined and used in 1974 by Norio Taniguchi in a paper titled “On the basic concept of Nano-technology”. And by the 1980s, people were regularly using and spreading the word “Nanotechnology”. Compared to large-scale counterparts, the nanomaterials tend to give better properties like increased strength, chemical reactivity, and conductivity. After entering the 20th century, the boom of nanotechnology has been enabling the wider and wider application of nanomaterials such as that in electronic devices, automobiles, cosmetic products, optics, even pharmaceuticals. (Hull & Bowman, 2014)
Nanoparticle pollution
The nanoparticles were defined as the ultrafine particles with a diameter between 1 to 100 nm. To put that in perspective, a nanometer is one-millionth of a millimeter, or about 100,000 times smaller than the diameter of a human hair. (“What is Nano? - The Nanodatabase,” 2013) It can be briefly classified into two categories, natural resourced-nanoparticles and engineered nanoparticles. It must not be overlooked that the nano-waste is generated from the process of the nanomaterial manufacturing process as well as from the after disposal nanodevices.
Generally, nanoparticles can be classified into six categories based on their fundamental structure in terms of carbon-based nanoparticles, metal-based nanoparticles, semiconductive nanoparticles, ceramic nanoparticles, polymeric nanoparticles, and lipid-based nanoparticles. (Ibrahim Khan, 2017) The diversity in chemical structure and property becomes one of the main obstacles in dealing with nanoparticle pollution.
Concerns about the health implications of nanomaterials have been expressed because particles and materials in the nanosize range may pose toxicological hazards due to their enhanced reactivity and potential for enhanced systemic availability. (Christie M. Sayes et. al., 2014) The release of the nanoparticle waste into the environment may generate ecotoxicity effect. Many studies indicate that exposure to the nanoparticles may lead to the absorption of ecotoxic nanoparticles and leading to irreversible damages. The mechanisms are various by different nanoparticle categories and the targeting organisms. (Boxall, Tiede, Chaudhry,2007) For example, an abundance of free chelating Fe nanoparticle was proved to have a close association with the lipid peroxidation, oxidative protein damage and DNA damage in living organisms. (M. A Shah, 2014) Furthermore, it has been reported that nanoparticle cytotoxicity in generating ROS could cause the mitochondria and nucleus damages in human cells. (Ana Lopez-Serrano, 2013)
In the water environment, nanomaterials are often more attractive to water and oil molecules, making them more absorbent than large-scale materials. In this case, nanomaterials were adopted in wastewater treatment to remove pollutants and to clean up oil spills at sea. However, the after used nanomaterials can not be completely removed from the water environment and might be aggregate and accumulate locally to cause long-term effects on local ecosystems. (Hull & Bowman, 2014)
Conventional Methods
The conventional methods have been classified into five main categories including sedimentation, interaction with biomass, specific membrane, continuous-flow system, and biological treatment. The drawbacks include but not limited to nanoparticle size, nanoparticle categories, and high cost in both economy and manpower.
Abstract
Based on such background knowledge and information, we decided to launch the iGEM2019 UM_Macau project SANCE focusing on nanoparticle pollution in the local water environment. We designed and conducted following human practices activities to establish the two-way dialogues between our team and the stakeholders with regards to the government, wastewater plants, and the public; seek enlightenment on the sustainability of SANCE, alleviate the concerns about GMO safety. The interviews, surveys, conversations with government authorities, and inter-team communications were conducted. As documented, we were able to complete the first two goals listed. As for the exploration of sustainability of SANCE, only the basic information was collected and the future direction was discussed. Through the whole process, we investigated the relationship between our SANCE and society with regards to how SANCE acts on nanoparticle pollution in the local wastewater treatment process as well as how the public reacts to applying E. coli as water treatment vector, and how the government response to our project.
Activities
Interview
In order to explore and investigate the real situation of nanoparticle pollution as well as obtain critical feedback from the associated stakeholders about GMO safety and possibility in the practical application of SANCE, we conducted the following interviews and surveys. The visit of WWTP broadened our understandings of current demands in Macau in the aspect of wastewater treatment and inspired us in designing SANCE. By contacting associated government authorities, we managed to drive more attention to nanoparticle pollution, which paved the way for SANCE application in the real WWTP process. Experts from associated areas gave us valuable suggestions, which motivated us informing further thinking in our functional tests stage and emerging new ideas of expanding sustainability by adopting SANCE.
We contacted the DSPA (Direcção dos Serviços de Protecção Ambiental DSPA, The official environmental protection bureau in Macau) and learned that there are currently five WWTPs in Macau. WWTP is considered as one of the stakeholders associated with our project. If we want to promote and implant our project into reality usage, the consultation with the WWTP is necessary. With the help of the government officers, we got a chance to visit the wastewater treatment plants in the Transborder Industrial Zone of Macau.
Figure 1. The group photo of members from UM_Macau and Pui Ching_Macau in front of the wastewater treatment plant.
From the introduction given by the plant’s manager, we learned the whole process of sewage treatment, from the collection to the evacuation. Four core stages were included in terms of pre-treatment, primary treatment, secondary treatment, and biological sludge treatment. We learned that currently, we don’t have the regular supervision systems targeting nanoparticle pollution. Besides, only the basic wastewater treatment techniques were adopted with unclear effects on nanoparticle clearance. The core technology applied in this plant is ATLANTIS® submerged Membrane Bio-Reactor(MBR) technology from Belgium company WaterLeu.
Figure 3. The presenting model of the MBR from WATERLEAU.
In general, the size of the pores in MBR is around 0.1-5 μm which is not able to clean up the ultrafine particles. The construction and operation cost of MBR is much higher than the traditional sludge treatment plant. (Le-Clech, Chen, & Fane, 2006) In the other four wastewater treatment plants, the most commonly used technology that may have effects on nanoparticle clearance is the secondary bio-sludge treatment which allows the bacteria in bio-sludge to clean up the pollutants in wastewater. However, the performance of this method is not satisfied when targeting nanoparticle pollution. Nanoparticle in wastewater processes cytotoxicity towards microorganisms, which could decrease the efficiency of sludge treatment. This informed us that the potential cytotoxicity generated by certain nanoparticles must be taken into consideration in the design of our own product. Under such circumstances, a more effective, economic, toxicity-tolerable method in targeting nanoparticle pollution is urgently needed.
We also learned from the manager that the microorganism control by UV exposure and chemical treatment in the late stage of the wastewater treatment process can achieve around 100% E. coli elimination. These results gave us faith in late GMO safety inspection.
Figure 4. The after-treated water is evacuated outside of the WWTP.
Aside from new product invention, it is also vital to draw more attention from the local government on the current issue of nanoparticle pollution. Therefore, we sent several emails to the related authorities including DSPA (the environmental protection bureau) and DSEC (the statistics and census bureau) under the government of the Macau Special Administrative Region. The related statistics in the quality of surrounding water was required, and the specific wastewater treatment technologies were enquired in emails.
We received the data and responds to our questions from all the authorities inquired. Unfortunately, the data are not allowed to be opened to the public. We noticed that though a vast amount of money was invested in wastewater treatment, nanoparticle pollution is still out of the concerns. While nanotechnology is rapidly developing globally, the associated regulations on nanotechnology applications as well as nano-waste disposal in Macau are still not well elaborated, and the nanoparticle supervision system is still not well established.
We then gave our recommendations with the proposition of calling for more discussions and official engrossments in the issue of nanoparticle pollution and urging the establishment of an efficient nanoparticle supervision system. It is reassuring that we got a very positive response from the government that more attention will be paid on the nanoparticle pollution as well as the related scientific breakthrough. The environmental protection bureau promised that they will keep focusing on the related issues to improve the water treatment facilities in Macau.
Figure 5. The flag of Macau.
At the functional test stage of our project, we noticed that there was a distance existing between our lab designs and the practical situation. The ideal sticky protein complex designed only based on original theory from labs may have difficulties in practical applications. We needed more professional suggestions in the knowledge of nanoparticle elimination for anticipating the potential risks in our design. Besides, more opinions from different perspectives were needed for better justification of our project. Therefore, with the help from our PI, we contacted three Hong Kong experts who are proficient in either nanoparticle or water environmental protection. By inquiries, we were able to have a broader vision in the nanoparticle industry and the possible drawbacks in our project, which inspired and motivated us to optimize our lab designs.
All professors presented their beliefs that the nanoparticle with exceed concentration is harmful to human health and the survival of certain microorganisms, which is consistent with what we originally learned and expected. As Professor Leung, Kenneth Mei Yee said,
“The current amount of nanoparticles is not enough to pose a serious threat to human health and marine ecosystem, but once its amount exceed a specified threshold, it could adverse human health”
Figure 6. Photo with Professor Leung, Kenneth Mei Yee.
However, we learned from the interview that the application of nanoparticles in real life is much wider than what we expected, which inspired us in the functional test stage to enlarge the categories of our targeting nanoparticles.
Some potential drawbacks in SANCE had been raised up by professors in concerns of low tolerance of nanoparticle cytotoxicity, which motivated us to improve our SANCE by maintaining the biological activity of our sticky protein complex even after the death of the organism itself.
Other concerns raised were mainly about the practical application of SANCE. The complicated interaction between activated sludge and nanoparticles adds more difficulties in predicting and controlling the movement and performance of SANCE. Therefore, Dr. Olivier Habimana did not recommend to applied SANCE in activated sludge, instead, he suggested we apply our product in the form of biofilm. With further consideration from this advice, we discovered the potential in SANCE of being applied as membrane treatment. The magnet feature of SANCE enabled the controllable immobilization. Together with suitable filter membrane as a vector, the biofilm system targeting nanoparticles enabled by SANCE is possible in practical application.
Figure 7. Photo with Dr. Olivier Habimana.
Moreover, the concerns in the safety of directly releasing SANCE as GMOs in wastewater treatment plants were addressed. Upon this issue, we recontacted the manager from WWTP in Macau and found out that the current E. coli elimination rate is very high up to 100%, which ensures no emission of GMO E. coli into the natural environment. Besides, the extra accessional magnetic field also gives double insurance in the control of SANCE from emission.
In other possible applications of SANCE, Professor V. Thiyagarajan provided another creative suggestion that SANCE can be used as a nanoparticle detector. It inspired us and open the possibility of applying SANCE not only in wastewater treatment but everywhere in the presence of nanoparticle pollution.
Figure 8. Photo with Professor V. Thiyagarajan.
Furthermore, several new techniques in detecting the concentration of nanoparticles were suggested by the experts. Fluorescence Correlation Spectroscopy (FCS) and AFM Force Spectroscopy can be adopted in not only detecting the concentration of nanoparticle but also the interaction between our adhesion system and nanoparticles.
Survey
Online Questionnaire
As a coastal city surrounded by ocean on the three sides, the life of Macaunese is tied up to the sea. Nanoparticle pollution in the surrounding water may directly affect the health of local residents. Therefore, it is crucial in the implementation of water-protection related policy that the basic knowledge of water pollution and protection are well spread and comprehended. Moreover, in order to promote our project in preparing the future application, to hear the opinions from the general public is one of the primary goals. In these concerns, we designed an online questionnaire with questions investigating how residents recognize water pollution and broadening their knowledge of nanoparticle pollution. We were able to collect about 251 valid replies at the end. The answer indicates that most Macanese have concerns that water pollution may influence their life and they are waiting for further water quality improvement in the surrounding water area. We realized the necessity to devise a product for helping to alleviate nanoparticle pollution in the local water area. The online survey indicated that local residents generally support the water improvement project, which provides more possibilities in the practical application of SANCE.
iGEM Team Communication
During the whole iGEM period, we proactively participated and initiated several inter-team communications including participating online and offline symposiums, contacting for team meet-up and lab collaborations. We received a lot of valuable suggestions in various perspectives and stages of our project with regard to iGEM regulations interpretation, project construction, and optimization, the past experience in Giant Jamboree.
With the introduction by our PI, we had a contact with the iGEM2019 team Hong Kong_CUHK and organized a meet-up in HK. This is the 7th time that the Chinese University of Hong Kong forming teams and participating in iGEM. The meet-up brought us a valuable chance of learning experience and exchanging ideas. With their assistance, we had a more distinct target in track selection. Their advice on plasmid construction also helped us in generating better protein expression. By listening to their sharing, we learned some skills and tips in participating in the final Giant Jamboree.
Figure 12. Group photos with members from Hong Kong_CUHK
After designing the bio-bricks, more perspectives and opinions were needed to justify our construction. We attended The Second Guangdong-Hong Kong-Macau Great Bay Area iGEM Synthetic Biology Development Forum. This forum attracted iGEM teams all around to share their ideas with each other through presentations and poster displays. We got some suggestions from other teams in experiment design and lab management. A good relationship and possibility of future lab collaborations were built up with the efforts of all related parties.
Figure 13. Photos in Guang Zhou symposium.
Held by TAC_Taipei, we joined the iGEM x Sustainable Development Goals Online Conference in order to discuss and extend the value of iGEM projects under consideration of the 17 UN sustainable development goals. The different targets and advantages of each team were presented related to one specific or more than one goal of the 17. We exchanged our views on GMO safety, different water quality regulations from different countries and regions as well as our efforts on promoting and assisting SDGs through the construction of our own iGEM project. As a team targeting the water environment protection, we mainly put our focus on the 6th and 14th goals on the webinar in terms of conserving and sustainably using the oceans, seas and marine resources for sustainable development.
Figure14. The screen-shot of the chat room of SDG & iGEM webinar.
In order to enhance the communication with the local teams to seek collaboration and learn experience, we participated in the 1st HK iGEM Symposium 2019 held by CUHK iGEM2019 team. Here we got a chance to present and illustrate our project to other four university teams and four high school teams from Hong Kong. With the advice and suggestions provided by other teams and PIs, we had a better understanding of basic iGEM regulations and acquired more experience in presenting and defending our propositions. This meeting also inspired us to establish the mentorship with local high school iGEM2019 team Pui Ching_Macau.
Figure 15. Photo of the 1st HK iGEM Symposium 2019.
Achievement
The design of SANCE is targeting the nanoparticle pollution in local wastewater. We designed and conducted the stakeholder analysis to investigate how does our project affect society and how does the local society respond to our project. Under these concerns, we conducted the human practices activities contacting the stakeholders with regards to the public, the government, and the wastewater treatment plant.
Through the online questionnaire towards the Macau residents, we found that there are a large number of replies believing that the water environment in Macau is not satisfied and more technologies need to be applied for more efficient treatment. In this way, we were able to hear the voice of the public and understand the current demands of water treatment. Besides, we also learned that most of the responders can accept the application of engineered microorganisms in the wastewater treatment process. The results surprised us that the public acceptance of GMOs in practical application is quite generous.
The interview with the WWTP manager enlightened us a lot on how to apply our SANCE in practical use. We then came up with two strategies for implementing the SANCE into the current wastewater treatment process. The first one is directly applying SANCE into the biological secondary active sludge in the primary treatment stage. However, this proposal was abandoned by us after the interview with the HK experts. There are too many components in sludge and the performance of our SANCE can not be predicted and controlled easily. Therefore, we gave the second proposal, applying SANCE after the sludge treatment before the sterilization stage. In this way, we can predict the movement of SANCE more easily as well as assess the overall performance of SANCE. Besides, separation from the sludge can also help with the controlling effect generated by the accessional magnetic field.
The contact with authorities under the government revealed the possible gaps in the regulations of nanotechnology and GMO safety. We then gave our recommendations upon the related issues to the government and got the positive replies from the government that more attention will be paid on the related questions and more encouragement will be provided to the emerging technologies upon wastewater treatment.
As for our GMO safety part, we learned from the government that currently, Macau does not have any other specific labeling regulations for GMO. However, concerning that applying GMO products in the real wastewater treatment process may lead to GMO contamination to the surrounding environment, we proposed a GMO contamination controlling system by combining the current sterilization stage of the WWTP in Macau with the accessional feature of our SANCE. After we discussed our concerns about the GMO contamination to our lab, we came up with this new feature of SANCE that by overexpressing the gene Ftna to magnetize SANCE by triggering the Ferric ions crystallization inside the E. coli. The expression of Ftna gene is able to generate ferritin protein complex and achieve the storage of Ferric ions. By adding the accessional magnetic field, we are able to achieve the immobilization of SANCE and avoid the release of the GMO product.
Figure 16. The concept diagram illustrates how is immobilization achieved. And the result presents the performance of our SANCE.
As for exploration of the sustainability of SANCE, we only had enough time for basic information collection and gave the primitive suggestion in future direction accordingly. Through the SDG & iGEM webinar, we had a chance to explore the potentials of SANCE in encouraging the development and advancement of the 17 Sustainable Development Goals.
By targeting the nanoparticle pollution in the local wastewater treatment process, the SANCE might be applied to help with the 6th Goal “Clean water and Sanitation” and the 14th Goal “Life below water”.
Acknowledgement
We would like to express our most sincere gratitude to those who assisted and helped us in this project SANCE by UM_Macau iGEM2019.
Hong Kong Experts
Professor Leung, Kenneth Mei Yee
Professor at The University of HongKong, expert in marine ecological and environmental science, provided knowledge of nanoparticle in the marine environment and comments on our project.
Dr. Olivier Habimana
Professor at the University of HongKong, recurrent research in the ecosystem and environment area, provided technique suggestions and his opinion about nanoparticles and SANCE.
Professor V. Thiyagarajan
Professor at The University of HongKong, an expert in marine ecological science, provided his opinion about the impact of nanoparticles on humans and the marine ecosystem and inspired us on other applications of SANCE.
The fishermen from sea products markets in Macau
Local fishermen from sea products markets called Mercado Municipal do Patane and fish stores next to the wharf, provided valuable and helpful information for our project.
Volunteers
Volunteers from the Faculty of Health Sciences, University of Macau undergraduate students assisted us in the seafood market interview in recording information, asking questions and taking photos.
Name List
- CHAN NGA IENG
- SU HONGYI
- FU IOI
- FENG YUZHAO
- XIE SHU YAO
Reference
- Taniguchi, N. (1974) On the Basic Concept of Nanotechnology. Proceedings of the International Conference on Production Engineering, Tokyo, 18-23.
- Boxall, A. B., Tiede, K., & Chaudhry, Q. (2007). Engineered nanomaterials in soils and water: how do they behave and could they pose a risk to human health? Nanomedicine, 2(6), 919–927. https://doi.org/10.2217/17435889.2.6.919
- Khan, I., Saeed, K., & Khan, I. (2017). Nanoparticles: Properties, applications and toxicities. Arabian Journal of Chemistry. https://doi.org/10.1016/j.arabjc.2017.05.011
- López-Serrano, A., Olivas, R. M., Landaluze, J. S., & Cámara, C. (2014). Nanoparticles: a global vision. Characterization, separation, and quantification methods. Potential environmental and health impact. Anal. Methods, 6(1), 38–56. https://doi.org/10.1039/C3AY40517F
- Sayes, C. M. (2014). The Relationships among Structure, Activity, and Toxicity of Engineered Nanoparticles. KONA Powder and Particle Journal, 31(0), 10–21. https://doi.org/10.14356/kona.2014002
- What is Nano? - The Nanodatabase. (2013). Retrieved October 19, 2019, from http://nanodb.dk/en/what-is-nano/
- Hull, M., & Bowman, D. (2014). Nanotechnology Environmental Health and Safety: Risks, Regulation, and Management. Retrieved from https://books.google.com/books/about/Nanotechnology_Environmental_Health_and.html?id=3ztOAwAAQBAJ&source=kp_cover
- Shah, M. A. (2015). Nanotechnology Applications for Improvements in Energy Efficiency and Environmental Management. Advances in Environmental Engineering and Green Technologies. https://doi.org/10.4018/978-1-4666-6304-6
- Le-Clech, P., Chen, V., & Fane, T. A. G. (2006). Fouling in membrane bioreactors used in wastewater treatment. Journal of Membrane Science, 284(1–2), 17–53. https://doi.org/10.1016/j.memsci.2006.08.019
- Yuen, C. (2018). Macau Food and Agricultural Import Regulations and Standards Report. Retrieved from https://www.fas.usda.gov/search/Macau
- Westerhoff, P. K., Kiser, M. A., & Hristovski, K. (2013). Nanomaterial Removal and Transformation During Biological Wastewater Treatment. Environmental Engineering Science, 30(3), 109–117. Retrieved from: DOI:10.1089/ees.2012.0340