At the beginning of our project, we visited local and foreign wastewater treatment plants (WWTPs) and learned that there are currently very few wastewater treatment methods that specifically target nanoparticle (NP) waste. We designed our constructs, prototype and modeling based directly from information given by these treatment plants. Our biofilm parts collection was designed to be able to control biofilm production in sedimentation tanks. Our proteorhodopsin construct is designed to be used in aeration tanks where other microbes are already breaking down organic substances. We also use the pre-existing biosafety aspect of the wastewater plants. Additionally, we reached out to several NP manufacturers, researchers, disposal services and wastewater experts who provided us with information on the advantages and potential consequences of NP usage, as well as where to target NP waste removal. The feedback we received guided the direction of our project and confirmed the importance of our project to our community.
Bioethics Panel
We hosted a Bioethics Panel, where we invited students and teachers to discuss the moral, social and environmental concerns of our project. To encourage participants to consider the problems from multiple perspectives, we created a role-playing game and assigned different roles to the participants. We then asked for their opinions on NP usage and disposal from the perspective of their assigned role. (Whole team activity)
For instance, one of our questions was:
“Dihua WWTP has no nanoparticle removal plan. Should this be the job of the wastewater plant? Or the nanoparticle manufacturer?”
The following roles were assigned:
Wastewater plant manager
Nanoparticle manufacturer
Citizen
Fisherman
Fish
Most of the wastewater plant managers thought that NP manufacturers should be responsible for removing NP, because they have more information (e.g., solubility, toxicity, etc.) about their own products. However, many other participants were skeptical that manufacturers could be trusted to remove their own contamination and agreed that WWTPs should ultimately be responsible for cleaning water contaminated with NPs.
This activity gave us great insight on how the public perceives NP usage and regulation, and provided us a chance to inform people about both the benefits and the dangers of using NPs. We first had the idea that we should create a filter that the consumers can purchase to clean NPs out of their household waste. However, after the bioethics panel, the results showed that most people think wastewater treatment plants functioning under the government should be responsible for cleaning NP waste. The reason being that all the wastewater would eventually accumulate in the WWTPs. Aside from analyzing the responses from the bioethics panel, we also tested products from NP manufactures (discussed below) that proved targeting wastewater would be the most ideal area to clean NP waste. This is why we decided to focus our project of nano trapping in WWTPs.
To learn more about the applications of NPs, we visited a nanotech company that uses silver NPs to make various antimicrobial products. The researcher and manager of Apex Nanotek, Chery Yang, introduced us to their main product, which is antimicrobial nanosilver activated carbon. Pure activated carbon, commonly used to treat sewage and industrial exhaust, is prone to bacterial growth. To overcome this problem, they integrate crystallized nanosilver into the activated carbon for their antimicrobial effects. One of their products is a showerhead, with nanosilver activated carbon filters to kill bacteria when water flows through the showerhead.
We tested the product by comparing SEM images between tap water and filtered water from the showerhead. The showerhead decreased the number of bacteria and larger particles from tap water! However, we also observed the release of NPs from the filter, which will flow into wastewater. (Interviewed by Christine C., Kelly C., Yvonne W., Chansie Y., and Justin Y.)
Chery Yang (third person from the left), the main researcher of Apex Nanotek Corporation
Product of Apex Nanotek: Silver Spring Shower Head.
Tap water under SEM.The image on the left shows a tap water sample under the SEM, in which we observed some bacteria (round objects that are approximately 1 μm in diameter). The SEM image on the left shows water that was filtered by the showerhead from Apex nanotek. There is less bacteria as the showerhead uses embedded nanosilver antibacterial filters. (SEM images: Christine C. and Florence L.)
WWTP -- Dihua Wastewater Treatment Plant
In order to learn firsthand about the effect of NPs in WWTPs, we visited the Dihua WWTP (迪化污水處理廠). Here, we were given a tour around the plant and were able to ask questions to the managers and people that work there. They confirmed with us that the current facilities are unable to remove NPs from wastewater mainly due to their small size. In addition to this information, they kindly provided us with samples of sludge, effluent water, and the flocculants they add during wastewater processing.
We plan to add our bacteria either in the deep aeration tanks or the secondary sedimentation tanks. The disinfection tank will kill the bacteria used in previous tanks.Figure: Christine C.
Throughout the year we visited and talked to the Dihua WWTP several times about where and how our project could be implemented in their current system. For our proteorhodopsin construct, we planned to add it in the deep aeration tank where the water flow is turbulent. Our biofilm will be attached to biocarriers that will then be placed into the sedimentation tanks (shown above). We inquired about the flow rates, dimensions, and time spent in each tank to see if they were significantly different, since we would be adding different construct designs. These conversations and visits played a huge role in shaping our construct design, prototype design, mathematical modeling and overall purpose for our project. For example, during our conversation with WWTP engineers, we learned that wastewater is retained in aeration tanks and sedimentation tanks for up to 4.8 and 3.8 hours, respectively. This means PR and biofilm will only have up to ~5 hours and ~4 hours respectively, to interact and trap NPs in wastewater; we adjusted our experimental design, prototype, and modeling to show our project working under realistic conditions. For instance, to arrive at a realistic expectation of how fast PR traps citrate-capped nanoparticles (CC-NPs), we added PR to CC-NP solution and mixed the solution for only up to ~5 hours (figure 2-6). We also added a flocculant powder (supplied by Dihua WWTP) to aggregate suspended solids to mimic the next step in the treatment process.
Figure 2-6 Proteorhodopsin traps CC-AgNPs. A) Absorbance decreased markedly when PR bacteria was added to CC-AgNPs; the absorbance did not change significantly when GFP-Gen (negative control) bacteria was added. B) Over the 5 hour period, we observed progressively larger dark orange spots (aggregated CC-AgNPs) in the PR group. Experiment & Figure: Justin Y.
Furthermore, we created a calculator based on our model that considers the aforementioned time limits—amongst other important variables, such as wastewater flow rate, bacteria concentration, and biofilm surface area—to calculate the amount of PR or biofilm necessary to trap the most NPs within ~4 hours and ~5 hours, respectively. Lastly, to test PR and biofilm in realistic conditions, we constructed a simulated WWTP (shown below).
We plan to add our bacteria either in the deep aeration tanks or the secondary sedimentation tanks. The disinfection tank will kill the bacteria used in previous tanks.Figure: Christine C.
Boswell Wastewater Treatment Plant
Not all WWTPs are as large as the one in Taipei. One of our advisors (Jude Clapper) went to visit the Boswell WWTP in rural southwestern Pennsylvania. We learned that the same processes that occur in the Taipei Dihua WWTP also occur in the Boswell WWTP, but with different water flow rates and waste quantities. Because of the similarities in how both WWTPs process their wastewater, It inspired us to create our current prototype design that is a rotating polymeric bioreactor coated in biofilm. This prototype will be placed in the secondary sedimentation tank, where the majority of organic solids have been removed and only smaller particles exist. The Boswell WWTP also confirmed that since our project is bacteria-based, it will be killed by UV light and chlorine in the disinfection tank, similar to the Dihua WWTP, before the water turns into effluent and goes to the rivers and oceans.
We plan to add our bacteria either in the deep aeration tanks or the secondary sedimentation tanks. The disinfection tank will kill the bacteria used in previous tanks.Figure: Christine C.
Thomas J. Brown
Thomas J. Brown, the Water Program Specialist of the Pennsylvania Department of Environmental Protection (DEP) occasionally helps with the Boswell WWTP. He has also worked with the EPA in Taiwan on wastewater treatments. We interviewed Mr. Brown about our methods to clean NPs in WWTPs and how to achieve our goal of implementation. With his expertise in the field of wastewater treatment, he provided us some suggestions as to how we could turn our project into reality.
For example, we asked him if there were differences between rural and urban plants that we should take into consideration when thinking about implementing our project. He responded that the biological processes used for treatment remains the same regardless of facility size. This helped us think about and design our final prototype, which can potentially be used in both rural and urban treatment plants.