HUMAN PRACTICES
Beginning our Journey: Identifying the Problem
Our university is situated in the vicinity of the railway station,
Katpadi Junction, which is a major source of PET bottle pollution, in
the form of Rail Neer bottles supplied by the Indian Railway Catering
and Tourism Corporation to train travellers. Our visits to the Katpadi
railway station made us acutely aware of this problem.
PET bottles were sold at station platforms and carried along by
passengers. Despite disposal systems being in place, the bottles were
still thrown around carelessly. We noted that a shredder-based system
had been installed at the station by the railway authorities, however it
was in complete disuse. A similar situation is seen in most Indian
railway stations.
![](https://static.igem.wiki/teams/4625/wiki/img-20230721-wa0052-1.png)
![](https://static.igem.wiki/teams/4625/wiki/img-20230721-wa0054-1.png)
Our interaction with a shopkeeper, Jeetu Bhaiya, revealed that roughly 500 PET bottles were sold at the station everyday. To further investigate the extent of plastic pollution, we visited places near the station, collecting samples at the Palar river bank, and nearby bus-stand. We observed several heaps of garbage dumped around these places, contaminating soil and water. We also found evidence of animals consuming plastic waste at these sites. We also noted other potential sources of pollution, from nearby textile industries and washing clothes in the river. We were also informed about a recycling unit near the Palar River that made slippers from recycled plastic.
![](https://static.igem.wiki/teams/4625/wiki/img-20230721-wa0058-1-1.png)
![](https://static.igem.wiki/teams/4625/wiki/img-20230721-wa0031-1-1.png)
![](https://static.igem.wiki/teams/4625/wiki/img-20230721-wa0043-1.png)
![](https://static.igem.wiki/teams/4625/wiki/img-20230721-wa0041-1.png)
To arrive at the crux of the problem, we consulted Dr Amitava Mukherjee, an expert on the effects of microplastic toxicity on algae. He informed us about the ramifications of microplastic leaching and pollution from these PET bottles, and encouraged us to investigate their impacts on human health, flora and fauna. We conducted a survey to gauge microplastic awareness amongst people, and understand their reasons for using PET bottles. Our survey received 200+ responses.
Quantifying levels of microplastics in tap water: A Global Initiative
Microplastic contamination is prevalent around the world. Realising the
global scale of our problem, we collaborated with iGEM Stockholm to
measure microplastic levels in tap water samples worldwide. We reached
out to various iGEM teams working on plastic and water pollution. We had
five participant teams - UNILausanne team from Switzerland, Makerere
team from Uganda, and Synfronteras team from Brazil, along with Team
Stockholm and our team. We opted for a simple protocol for microplastic
detection using Nile Red/Rose Bengal staining. Our aim was to create a
standardised protocol that could be performed quickly, and with minimal
equipment.
Although we failed to generate conclusive results, through our efforts,
we were able to understand the need for better and more convenient
microplastic detection methods. We also explored the possibility of
using a fluorescence based method for microplastic quantification.
However, due to time and resource constraints, we were unable to carry
out the experiment.
- Team VIT-Vellore
- Team Stockholm
- Team Makerere-Uganda
- UNILausanne
- UNILA-LatAM (SynFronteras)
We performed the Nile-Red staining method, observing the stained and unstained regions of the filter paper under high power of the microscope (40X).
![](https://static.igem.wiki/teams/4625/wiki/t76kt6bjee9ea30h0zmk4rdrw0npmxddsxqgm51e5r3lntunj4leitnhr-1jdodvj-7u-6q104a4vq5agdsc-uchpxs2orauwt0efefbvn0-z1orgae7m64-hfjhutxzkf4fhbkscanwqxenfdpd5y-1.png)
![](https://static.igem.wiki/teams/4625/wiki/m3vtdfo9fh5ixfzvdvv2b0uuxbxlpwzsvt8wb-mkjx-qwhux-xcw0wq1veqo0r3fg92mwyhzlndi3n42zjmjcrdxogwxyu2muerleyeszb-ogwdjfdkzgqq97pdfbq7ryk3yz27-m-bv2hwf0alixe-1.png)
The team researched on FTIR measurement of microplastics and approached experts to devise a protocol and test it out. A lack of time hindered them from performing the experiment.
The team performed the Nile-Red staining experiment, and were advised to optimise the protocol further by their project investigator to get better results.
![](https://static.igem.wiki/teams/4625/wiki/tv09mjd2kq9jafdkucrk0uwhm0fhqqn9e6fgzuyu9ipmdqae6-aimupzydokzwy98ezkzv4lbieqhqkdl3oqu1mqsey56cudeh9pfcrkghtlfs2azdeh7qeyi0euhp9n0rzgchq23patg0k1zlsqafy-1.png)
![](https://static.igem.wiki/teams/4625/wiki/pwv-o4ljkh7wij9wrwsoiio47jdnmruxpj2ckjf6fxyn6fvqd5rbvmoi-ubeme9d-t53dnj-eppebbupvcztolw-0y4y-b6k0p8ru8p0ephl-esg4qkv8jfisgpzyfmmf3rbstvycxgn99x30d6qo-1.png)
The team was able to perform the Nile-Red staining experiment, however faced challenges due to extrinsic contamination of their water samples. They were unable to resolve this problem, even after re-performing the experiment.
![](https://static.igem.wiki/teams/4625/wiki/0zhhpwtsmuiz-pdm-j-67kpzxh-vzi1dfvznhyo-jdqr6th66il-8enqz1oqeyzhff5k5oeppandif7xvs37-w1foj2xhohddcffgbjsz1ltuajurb8hjmbq7rauifmxog1hotmcih10smwyai42wu-1.png)
![](https://static.igem.wiki/teams/4625/wiki/2j2kom-ymadls7kpzfvbchwzfxcs08-pnzuketggbrtvfnpwsxttb9r9e2l-ubwdpdwcthvjjvlu0-ebvjlyz3-sshic6-1izy-cbayr2ymhbasnatieuntzllxq-tu62ldah2c9o-dawcsqjz7uw-1.png)
The team faced challenges procuring the required stains, and filter paper. They were unable to obtain clear results, but labeled possible areas of detection.
![](https://static.igem.wiki/teams/4625/wiki/kazmvphxwi71l-xpiowehdrv-xl0t6fijofl8i5wasnvpokqwiu2bkkv-nxd52qdlezaxuswvyat7ytx05gnfcseqbxfhx2hzxun0bnmrc0kcca26sb6rc3-vlm47txn4sr8coyjwtxjxohvwxpatou-1.png)
![](https://static.igem.wiki/teams/4625/wiki/0spr8npq3a5syenkwxt2-fqfr2jofh-2lq-kypn5sxxuywk8ssomoylqztkdyvly5clcgcihi1iwc-xhxsqqwrllc26q9qhbrikwyp2pvojsax1ez-4le3viwu08hknhhytpc9ghvylyttcejchcdum-1.png)
This initiative proved to be a great learning experience for us, providing us valuable insight on the drawbacks of current microplastic detection methods and the ease with which they can be performed. With eager participation from five teams across four continents, we were able to generate awareness of microplastic contamination at a global scale.
Dry Lab Inputs
For developing a mathematical model for our enzymatic system, we approached Dr. Sajitha Lulu. She guided us on conducting a literature survey on wild type PETase and MHETase and working on an E-cell and V-cell model for checking the efficiency of our enzyme in real time. We were further encouraged by Dr Saravana Prakash Thirumuruganandham to conduct in-silico analyses such as Normal Mode Analysis, non-linear normal mode analysis, and quasiharmonic simulations to increase the novelty of our project.
![](https://static.igem.wiki/teams/4625/wiki/whatsapp-image-2023-09-27-at-12-16-1.png)
![](https://static.igem.wiki/teams/4625/wiki/whatsapp-image-2023-09-27-at-12-12-1.png)
Finding the best fit for our solution: Railways or Recycling Units?
In order to discuss possible implementation strategies for our dual enzyme system, we met Ms. Arjitha, Health Inspector at Katpadi Junction. She informed us that incorporating the enzyme-based system into the pre-existing shredder would be challenging at this stage, considering the state of disrepair that the shredders are in, and other infrastructural limitations. She also suggested that in order for the shredder-based solution to be effective, the behavioural tendencies of people need to be tackled - they need to be made more aware of the correct disposal methods for PET bottles. To gain a better understanding of this behaviour, we performed a random sampling of 20 train passengers - asking them if they reused PET bottles, and whether they were aware of the “crush after use” instruction on the bottle, and their methods of disposing of the bottle. Most people told us that despite being aware of the instructions, they did not not adhere to it.
![](https://static.igem.wiki/teams/4625/wiki/pasted-image-0-1.png)
![](https://static.igem.wiki/teams/4625/wiki/pie-4-removebg-preview-1.png)
![](https://static.igem.wiki/teams/4625/wiki/pie-2-removebg-preview-1.png)
Ms. Arjitha also brought to our notice several campaigns that the
Indian Railways and the government have conducted in the recent past
to mitigate this problem. To assist them in their endeavours, in
collaboration with the Indian Railways, and VIT University, our team
organised a clean-up drive at the Katpadi Station. This event saw the
enthusiastic involvement of many students and faculty members from our
university.
We considered another potential avenue for implementation at recycling
and waste-segregation units. Our discussions with Mr Manil, Founder of
SustEnable, revealed to us that municipal corporation workers could be
a good target demographic for our solution. We visited the Vellore
City Municipal Corporation Composting Shed, where Mr Srinivasan walked
us through the process of recycling. First, the waste was segregated
into wet and dry, and plastics and papers were separated. The
biodegradable waste was used as compost for agriculture, and the
non-recyclable plastics were stored as solid waste in a godown near
the Palar river. Interestingly, we also observed that several PET
bottles were used in the construction of walls at the segregation
unit.
![](https://static.igem.wiki/teams/4625/wiki/img-20230730-wa0037-1.png)
![](https://static.igem.wiki/teams/4625/wiki/img-20230723-wa0021-1.png)
During our online seminar in collaboration with Stockholm, we interacted with Dr Prassana Venkhatesh, Senior Research Analyst (Sustainability Program) at STEER Engineering Pvt Ltd, a corporation working on bioplastic alternatives and plastic recycling. Through his informative presentation, we learned about the possibility of incorporating our enzymatic solution to industrial bioreactors. The conference also saw the attendance of high school student researchers- Lucia Royo and Ainara San Miguel, researchers and academicians - Dr Hoi Shing Lo and Dr Amitava Mukherjee, and Dana Dedeck, a program official involved in water governance from the Stockholm International Water Institute (SIWI), and provided a holistic overview of the microplastic problem.
![](https://static.igem.wiki/teams/4625/wiki/me41ucj44xb4omkjcxs4zfywjgxuvv8tkjx1ph4to9-24bbwpula1vzm6au0ygpm4q-zfwd0wzrqwpbargkw5veaumqf2nugby7ta2lcodc1bsd9bcayodlyvl1swuwvyvn-sq5km3vcmh1mrhzl1ty-1.png)
What are the stakes?: Incorporating diverse perspectives
At the All India iGEM Meet, Lavanya Ma’am, Founder and CEO of Anvaya Biotechnology Ltd., suggested that we focus on sustainability, and interact with stakeholders such as policy makers, non-governmental organisations and municipal corporations.
![](https://static.igem.wiki/teams/4625/wiki/pasted-image-0-2.png)
Taking her advice in consideration, we have reached out to the major groups and other stakeholders associated with our project - Business and Industry, Children and Youth, Local Authorities, Non-governmental Organisations, Scientific and Technological Community, and Education and Academia. Integrating their inputs at every step of the process has helped us clearly define our problem statement, circumvent any technical difficulties we encountered, understand the socio-economic grounds our project stands on, and consider potential arenas for its implementation.