Human Practices

Introduction

Human Practices was a way to steer our project towards social responsibility and realism for us as a team. Through our project, we hope to contribute to the resolution of the global problem of lead poisoning. Inspired by the approaches used in responsible research and innovation, we combined the ELSA approach and AR to anticipate issues that may arise from our project's large-scale implementation in the future. We ensure that the insights gained from this are addressed significantly in the current implementation of our project. Our human practices initiatives were used not only to guide our project, but also to aid in the relief of lead poisoning in India even beyond the project, truly embodying the spirit of "locals solving local problems."

Thus, human practices were primarily used to gain insights into our project's technicalities whenever we felt stuck.
1. Obtaining insights into our project's ethical, legal, and social feasibility.
2. Gaining insight into the problem of lead poisoning in India and the general public's perception of it
3. Gaining insights into potential solutions aids in the resolution of the lead poisoning problem outside of synthetic biology.
4. Gaining insights into the progress of synthetic biology in India and future steps to make synthetic biology more democratic and responsible

While investigating lead poisoning as a problem and assisting in its resolution through synthetic biology, we discovered that:

1. Detection of lead in samples is a problem in and of itself:

The CDC (Centers for Disease Control and Prevention) changed the blood lead reference value from 5 milligrams per decilitre to 3.5 milligrams per decilitre in October 2021. This was the first change in the reference value in nearly a decade, implying that Lead affects an increasing number of children. Furthermore, we discovered that current lead detection methods are performed in high-end labs, only centralizing the process and delaying decision-making by governing bodies to prevent lead poisoning. As a result, we concentrated on resolving this issue by proposing a biosensor that can be accessed locally to quickly quantify lead at a decentralized level. Biosafety concerns were raised with our biosensor during the course of our project, so we proposed using biosensors for isolated samples in an isolated system. These isolated samples could be water or soil, two of India's most common sources of lead poisoning.


2. Dangerous lead levels in drinking water are used daily:

In most of the samples collected from water bodies across India, we discovered through a literature review that lead content in water is three to four times higher than BIS specifications. As a result, we proposed incorporating a lead recovery system without a project to assist in the recovery of lead from water samples. Again, we recognized a biosafety concern for this system, so we proposed using a bioreactor to ensure biocontainment of the engineered cells.


3. People in India are not aware of the dangers of lead poisoning:

We conducted a survey to assess IIT Delhi students' knowledge of lead poisoning. According to the survey, 2 of 10 people from India's top technological institutions classified 'lead as a nutrient.' The survey also revealed a general lack of knowledge about common sources of lead poisoning and the diagnosis and prognosis of lead poisoning. This inspired us to spread the word about lead poisoning and figure out how to approach stakeholders for actionable insights.


4. In India, there is a lack of understanding of synthetic biology:

We also conducted a Synthetic Biology survey at IIT Delhi and discovered that most people are unaware of the field and its implications. As a result, we integrated the synthetic biology education initiative into our human practices.

We proposed a technological solution for lead positioning: a biosensor and lead recovery system based on synthetic biology. However, the third and fourth points were impediments to steering our technocratic solution toward a democratic solution. This was because lead poisoning is a precursor to the majority of the diseases that people are concerned about. Furthermore, because blood lead levels are not tested in India, most affected people are unaware they are victims. Even if they were aware that they were affected, they have no idea why, and they simply ignore it because most jobs involving lead poisoning in India employ people from very poor backgrounds, so they prioritize their livelihood over their well-being. The general public's lack of awareness of Synthetic Biology is due to a lack of education in this field.

As a result of the aforementioned issues, gathering insights from the primary stakeholders affected was time-consuming and inefficient. We decided ELSA was the most suitable framework for our project because we wanted to investigate its potential ethical, legal, and social aspects, not just its implications as the ELSA framework may imply. We decided not to follow the AREA framework because they were unsuitable for our project. AREA relied on a level of engagement with end users that were unavailable to us since our primary stakeholders were unaware of lead poisoning.

So we decided to work around the problem by speaking with stakeholders and experts in the field of lead poisoning and synthetic biology. We identified experts in the following fields:

  • Public Health Policy
  • Occupational Health and Safety Policy
  • Policy on Synthetic Biology
  • Treatment of Wastewater
  • Biosafety

Interacting with these experts provides insights into how, in our case, implementing our project using the precautionary principle can help us create a larger impact in a shorter time frame. The precautionary principle was suggested by prioritizing the lead recovery system used in our project to recover lead from lead-contaminated water samples, developed on our IHP page (add the Integrated Human Practices link here). Also, there were interactions suggesting biosafety protocols and the economic value of our implementation. These interactions were used to incorporate insights into our project to have a broader and greater impact over the course of the project and in the future.

We used our human practices not only to steer the project toward social responsibility and realism but also to seek technical expertise whenever we encountered technical difficulties while implementing our technocratic solution. We contacted experts in the following fields on numerous occasions:

  1. Structural Modeling
  2. Mathematical Modeling
  3. Microfluidics
  4. Biosensors
  5. Bioreactors
  6. Synthetic Biology

These reach-outs were sometimes explicit, as when we contacted our institute's professors and seniors for immediate advice to unclog our pipeline whenever we were stuck. Sometimes our requests were implicit, so we asked experts to review our work/methodology and make any necessary changes to assist us in realizing and improving our project technically.

These are the key human practices initiatives we used to guide our project through its various stages.

Surveys

The survey's advantage lies in the simplicity with which it was designed to handle and quantify the data that we have recovered and the potential volume of responses. Even though it doesn't return in-depth answers like in personal interviews, if the survey is done well and receives a sufficient number of responses, it can still give a broad sense of what awareness the public has.

Intending to assess the current awareness about lead pollution among people, iGEM IIT Delhi conducted two surveys about lead poisoning. Survey questions were designed to achieve our goal to understand the current state of awareness regarding lead poisoning and contamination along with facilitating the spread of information about lead pollution, like the primary cause, effect, and testing methods for lead poisoning, through an interactive survey. This not only helped us assess but also educate at the same time.
While conducting the survey, we ensured that the participants had complete information about the subsequent use of the information gathered by the iGEM project at IIT Delhi. Additionally, it was anonymous to ensure the privacy of the respondents.

Domestic survey:

We conducted the domestic survey among the Indian public, with most participants being students between the ages of 18-22.

The survey can be viewed at: https://forms.gle/Qjgy9bZ1i24Jqrjj9.

The questions were designed taking lead poisoning statistics and sources collected from government websites and past research papers.
For this survey, we collaborated with the Board of Student Publications(BSP), the journalistic body of IIT Delhi , to reach out to the relevant participants and for subsequent distribution of the survey results to spread awareness. The survey link was also shared through our Instagram page and publicly displayed in our presentation at the All India iGEM Meet 2022 at the Indian Institute of Science, Bangalore.

The report of the survey was printed and published through BSP IIT Delhi. It can be viewed at : https://www.bspiitd.com/post/lead-poisoning-report-by-igem-iitd

Results from the survey:

The findings suggested a concerning level of ignorance in the community about the sources of lead exposure and the harmful consequences of its prolonged exposure. 93.2% of people have never encountered anyone who has undergone testing for lead poisoning. Only a tiny fraction of 4.5% of people have their water pipes checked, while most people are unaware whether their water supply pipes are leaded or not. This question was also accompanied by an image with the instructions to perform a simple test to check for leaded pipes to promote testing at an individual level. Since 35.3% of people never thought about paint as a potential source and 19.3% believed that packing food in newspapers cannot expose you to lead poisoning, there seems to be a serious requirement for spreading awareness about it.

Global Survey:

The global survey was a modified domestic survey conducted to understand this issue's perception and awareness in diverse methodologies and countries. To get global responses, we collaborated with the Office of International Programs, IIT Delhi, and also used the iGEM global slack channel as a platform to reach other international teams. Through this initiative, we understood the diversity of the issue and the importance of our project in different countries.

The Global Survey helped us understand the level of awareness of people in different countries and how safety with respect to lead poisoning was dealt with in their geographies.
This information helped us establish two things:
1. Identify areas of growth for awareness in the Indian Context and how other countries had dealt with this
2. To establish scope of iGEM IITD’s work in geographies other than the home location, i.e., beyond India

The information collected from the survey to better tailor our strategy for the project to the public and incorporate these messages into the project design. Additionally, it helped us gain a larger view on the applicability of this project and better isolate our use cases, and subsequently our market size as well.
Following this, we also set up an initiative to spread awareness through a social campaign on the IITD campus. In which we put up posters on awareness regarding lead poisoning in general areas to sensitize people regarding the issue.

Stakeholder Interaction

To make a societal impact with our innovation, we have to be aware of who our stakeholders are, what they value, and keep them involved in our project. Therefore, from the start of our project, we placed great emphasis on approaching, engaging and analysing stakeholders in the appropriate way.
Moreover, we ensured we were consulting as many relevant stakeholders as possible, from different backgrounds and who may have different opinions and perspectives. New information acquired along with our exchanges with stakeholders prompted questions such as: Does this change an aspect of our project? Is this revealing something we had not thought of before? Who should we consult in order to best answer our doubts?
Here, we describe our process of identifying and approaching our stakeholders.

Identification of Stakeholders

By identifying groups who have an interest in solving the lead poisoning problem or in our project itself, we have identified and engaged with stakeholders who may play a role in developing and implementing LEADer in the real world.

When deciding who to reach out to, we thought of the following parameters:

  • Expertise in our relevant scientific fields (synthetic biology, biosensors, heavy metal poisoning, heavy metal detection and bioremediation)
  • Expertise in the issue
  • Influence on the situation
  • Most affected by the issue

We thus elaborated the following list of stakeholders:

  1. Wastewater treatment plants would be the end-users of the product. LEADer is dependent on their willingness to incorporate our biosensor, or some version of it, into their facility, so that the lead-contaminated water can be treated. Their input on our implementation, as well as their knowledge of water treatment, is thus of great importance.
  2. The scientific community includes people from various fields of research and technical expertise. These include professors having areas of interest in biosensors, synthetic biology, wastewater treatment and lead metallurgy. By speaking to them, we were able to improve our project and hence constructed a more safe, more realistic and well-rounded project.
  3. Regulators and Policymakers are the ones responsible for dictating and implementing lead prevention policies in India. They are essential to help us grasp the legal and political frameworks in which we will build our project. Moreover, they usually have an overview of the situation, being themselves at the forefront of change.
  4. Lead-based Industries are the ones working primarily with lead. They include our project's major application. It was essential for our project to understand how these industries remove the lead waste that has been generated. Moreover, their inputs will guide us to make our product industry ready, by increasing our understanding of what these industries need.
  5. Public and its take on our project is essential. These include people affected by lead poisoning and the areas where lead contamination is an established problem. By knowing the problems faced by them in their local region, we were able to improve our product accordingly.
  6. NGOs and its perception of our project were crucial. These includes various NGOs (Non-Governmental Organisations) that were working on the same goal of making the environment lead-free. It was essential for our project to know how these groups work locally and what problems they face.

Reaching out to Stakeholders

Before engaging with any stakeholder, we developed a methodology based on our goals and needs. We needed a rigorous but adaptable framework that would allow us to understand the impacts of our project, and how we could make it better.
In order to determine these methodologies, we reviewed the projects of the outstanding iGEM teams in previous years for inspiration and make sure to adapt our methods to each situation.
We also conducted research on our side and discussed frequently amongst ourselves to evaluate our current methodology and ensure it was still responding to our needs.

We began by contacting our stakeholders through email. In this email, we introduced ourselves, our project, why we are interested in approaching that particular stakeholder, and also attached a project pamphlet. We asked whether they would be interested in sharing their insights in an online interview. Meeting online lowered the threshold to meet with people from different regions, it allowed us to reach out to stakeholders not only in New Delhi, but also, for example, in Bangalore and Maharashtra.

In addition, we asked for informed consent to share our insights from this interview, including the name and photograph of the interviewee on our wiki page.

While meeting on more theoretical matters such as biosensor simulation or lead poisoning policies could occur online, we found it essential to visit wastewater treatment plants in person, so as to truly grasp the contexts in which our project would live.
We tried reaching lead-based industries through email available on their website. But after getting no responses we changed our approach and tried to reach out to the employees of these industries on LinkedIn. But unfortunately, we were unable to get any replies.
By contacting numerous NGOs and social activist groups, we attempted to get in touch with lead poisoning victims. We were unable to identify any social activist organizations serving only the needs of lead poisoning sufferers in India, nevertheless.
Then, we adopted a different strategy, broadened our emphasis, and made an email-based outreach effort to environmental NGOs with a similar objective to stop heavy metal poisoning. We started making cold calls to them after receiving no response, but regrettably, no one answered.
As a result of these constraints, we find ourselves in a position where we are unable to develop meaningful relationships with those who are suffering from lead poisoning. However, iGEM IITD does recognise these individuals as significant stakeholders in our analyses.

To ensure our implementation was realistic, and to have an idea of the system it would be immersing itself in, we visited a wastewater treatment plant in IIT Delhi. During our visit, we held interviews with personnel there. He was able to inform us about the functioning of the facility and gave us an overview of the work they perform there.
Here is a summary of how the plant operates:

Sewage water first enters the receiving chamber via the MPS (Main Pumping Station), and then it passes through a fine screen filter to remove big objects and macroscopic particles like polythene. It next passes through a grit chamber, where heavy waste settles to the chamber's bottom

The oil and grease chamber, which is used to catch oil, grease fats, food solids, etc., is the next stop.

This water now enters SBRs (Sequential Batch Reactors), where the wastewater is aerated by having oxygen pushed through it by a diffuser. Bacteria and other germs settle and are referred to as sludge when the fan is turned off. (This sludge, created during the aeration process is transferred through a SAS (Surplus Activated Charcoal) pump into a filter press, where it is dried by passing air through it. This dried sludge can be utilised as fertiliser for plants.)

The water is then disinfected by passing it through the centrifuge decanter and into the chlorination tank. This chlorinated water is then filtered through a media, and UV disinfection is used to produce treated water.
This cleaned water is kept in a reservoir where it is delivered to IIT parks, gardens, fountains, etc., and used there.

Our project was strongly impacted by our interactions with stakeholders. Their advice on how to carry out our plan and their expertise enabled us to improve our project by bringing to light previously unconsidered factors. This encompassed not just the technical parts of the project but also leading our project to ensure that it was feasible.

Integrated Human Practices

The ethical, legal, and social approach, often known as ELSA, was chosen as the guiding principle for our project. People at risk of lead poisoning were designated as the primary stakeholders in our project. To orient our initiative toward social responsibility and reality, we consulted specialists in public health, occupational health, synthetic biology policy, wastewater treatment, and biosensors from all around India.

According to those knowledgeable in health and public policy, the first thing that should be done when there is a cause for concern (such as detecting high BLLs) is to implement the precautionary principle. Also, the inputs obtained by experts working on Biosensors were suggested in favor of lead recovery systems given the time scale of our project. As a result, we decided that prioritizing the lead recovery system would significantly impact our project in a shorter amount of time. As a result, we decided to prioritize our efforts in the wet lab on lead recovery systems over biosensors. Here, we do have to acknowledge that the Biosensor we are proposing is an essential device for effective decision-making and future research on lead poisoning. We also studied system utility preferences (Lead Recovery or Lead Biosensor). The former, which had a more comprehensive range of applications, resulted in a more significant agreement. The acceptability of our project has been ensured.

In the wet lab, we had support from specialists in India's policy regarding synthetic biology and biosensors. During our conversations, we concluded that using immobilized cells rather than microfluidics may make our biosensor more efficient in terms of cost and time, so we incorporated that. The suggested operation for treating wastewater will be more effective if it uses an immobilized cell technology. During our interactions, the experts working on synthetic biology policy made us aware of the implications of genetically engineered bacteria. They helped us guide towards effective proposed implementation in India because of their input on regulating the use of GMOs in India.



Dr. Arati Ramesh





Dr. Arati works at the National Centre for Biological Sciences in TIFR, India. Her primary work is in the class of non-coding RNAs called riboswitches, which directly bind cellular metabolites to control the expression of downstream genes.

Dr. Arati gave extremely insightful inputs with regards to safety, structural modelling and implementation of the project. In terms of structural modelling she advised that we needed to be extremely careful in interpreting our results when performing metal-protein docking, primarily because a metal ion by itself is an extremely small entity and current modelling softwares may not be that sophisticated to be able to accurately dock it with the protein and measure the binding affinity. Her rationale made us realise an improved strategy for molecular docking by measuring the best binding in terms of conformational stability of the complex after binding instead of trying to assess affinity. This helped us improve our understanding of current limitations and methods in structural modelling techniques.

In terms of the implementation, we also discussed progressing towards cell-free systems for the biosensor instead of a whole-cell system. She pointed out the crucial limitations of a cell-free system: even for the proper functioning of the machinery without the cell, it would require optimum pH, temperature, and salt conditions, among others. In the absence of a cell, these aren’t automatically regulated and could hamper the system’s functioning in the presence of environmental samples. On the other hand, whole-cell systems can regulate these conditions better, even in ecological samples, making them more reliable and stable. However, biocontainment is guaranteed in cell-free systems, unlike whole-cell ones. This tradeoff would have to be resolved by properly designing a contained environment like the microfluidic array for the biosensor.

She also posed interesting evolutionary questions regarding what kind of selection pressures this would put on our engineered E. coli , whether exposure to high concentrations of lead for recovery and detection could cause it to adapt to develop resistance to the heavy metal over time and what kind of implications this could have.




Dr. Krishna Ravi Srinivas:





Dr. Krishna Ravi Srinivas is the Managing Editor of the journal ‘Asian Biotechnology and Development Review’ and is affiliated with Research and Information System for Developing Countries. He has been a visiting scholar at the University of Pennsylvania, a visiting scholar at Indiana University, Bloomington, anda Post-Doc Research Fellow at South Center, Geneva. He has published extensively on intellectual property rights, climate change and technology transfer, open source innovation, and traditional knowledge.

He was extremely kind and motivating for our team, especially since he was the first stakeholder that we interacted with. He helped us understand the current state of synthetic biology in India and abroad and drew parallels and differences between the two since he had varied experiences. We discussed how synthetic biology is progressing in India and what we can do as students to promote and accelerate its growth. We identified that poor science communication is among the primary reasons for people, especially those who are not well-versed with life sciences, to be averse to the idea of synthetic biology. Terms like “unnatural,” often used in media to describe synthetic biology innovations, makes it seem like a scary unregulated affair. We discussed how it was essential to improve methods of communication of engineering biology along with stressing on discussions on bioethics and safety to help this field progress further. He reflected on our education and communication initiatives and appreciated the amount of feedback we looked for from our stakeholders as well as the students and teachers receiving our developed SynBio content.




Prof. Ravikrishnan Elangovan





Prof. Ravikrishnan Elangovan is an Associate Professor in the Department of Biochemical Engineering and Biotechnology at the Indian Institute of Technology Delhi. He works on single-molecule biophysics, fluorescence spectroscops, molecular motors, and skeletal muscle mechanics. He also has great experience in microfluidics, especially in the domain of biology.

He made us realize that the microfluidic device we wished to design for the biosensor would require a lot of precision as well as optimization and might not turn out the way we want it to in merely the first trial. Also, due to the unavailability of resources and limited time, it would be difficult for us to fabricate such a device. As an alternative, he introduced to us a very logical approach that might even turn out to be better than making a microfluidic device, i.e., immobilization of cells using calcium alginate / low melting agarose. He pointed out that all we need are cells trapped in a porous medium enough for the exchange of nutrients and gases from the surroundings, which would allow it to perfectly fit into our idea.




Prof. Sanjay Ghosh:





Prof. Sanjay Ghosh is a Senior Assistant Professor of Synthetic Biology at IBAB(Institute of Bioinformatics and Applied Biotechnology). His primary research is in developing translatable technologies and products using synthetic biology approaches. He works on various genome editing technologies and DNA assembly methods to engineer microorganisms, mosquitoes, and human cells.

Prof. Sanjay gave us extremely innovative suggestions, like using quorum sensing in our recovery system, apart from the biosensor. Forming a biofilm of lead adsorbing bacteria could be an exciting application of our system once we characterize its ability to interact with lead. We could use this to develop more efficient lead adsorbing strategies. He also suggested using a consortium of our lead adsorbing bacteria expressing different lead-binding proteins to test if a combination could outperform individual types of the engineered bacteria. These were among the most out-of-the-box problems we had thought about in our project as possible implementations of it in the future.




Dr. Shyam Pingle:





Dr. Shyam Pingle is a senior Occupational Health Physician in India who is passionate about Occupational Health, Workplace Wellness, and Community Health. Dr. Pingle has worked as Chief of Medical Services in various industries. He is an active participant in promoting and advocating occupational health in India and other developing countries.

He provided insights on health issues faced by people working in heavy-metal-based industries. He brought to our notice the shortfalls in current regulations. Currently, checkups for blood lead levels occur regularly, but they lack quality control and execution. Following this, we were suggested to work on a solution post lead detection, i.e., towards recovery systems for such industries. Dr. Pingle liked our project and invited us to present our work at OCCUCON (Occupational Health Conference), held in January 2023 in Delhi. The Indian Association of Occupational Health (IAOH) organized the conference.




Prof. Srikanth Mutnuri:





Prof. Srikanth Mutnuri is a professor of the Applied and Environmental Biotechnology Laboratory at the Dept. of Biological Sciences in BITS Pilani. His main research is on Sustainable Development Goals with a major emphasis on Clean water Sanitation, Zero hunger, Affordable & Clean Energy, Climate Action, and Responsible consumption and production. He had researched BLLs in extensive mining districts of India.

During our interaction with him, we explained our project, and he was deeply impressed by it seeing us leaping toward solving lead poisoning in India. He informed us of ignorance towards this sensitive topic and listed its reasons. One of the reasons he mentioned that centralized lead testing only in high-end labs is one of the key contributors to this ignorance. He told us that our Biosensor could go a long way to bridging this gap as it provides the decentralized basis for the quantification of lead which authorities can integrate with local wastewater treatment plants for water treatment for lead and make quick decisions. He also made us aware of the potential exposure in mechanical shops in India where lead acid batteries used in cars are handled improperly. He suggested using our lead recovery system in series with industrial treatments currently being used for lead treatment. He told us about the use of MBBR (Moving Bed Biofilm Reactor), which are being used in bioaugmentation.




Dr. Swathi Alagesan:





Dr. Swathi Alagesan is a Department of Science and Technology Fellow Faculty at IBAB (Institute of Bioinformatics and Applied Biotechnology). Her current research is focused on generating various synthetic biology tools and using the insights obtained from flux analysis to engineer microorganisms for optimising behaviour.

She has experience in developing a biosensor to detect Arsenic poisoning in blood. She says that metal recovery is the next big thing to work on, as a lot of work has already been done on detection. She believes that in the future, we could integrate a thresholding metal detection circuit with lead-recovery cells. This won’t increase the complexity like that of an oscillating sensor while also helping us achieve two objectives. Additionally, a circuit to express the short domain of PbrR as the surface protein for lead detection could also help improve efficiency by reducing the bacterial load. Initially, we planned to use heat shock to weaken the lead-protein binding and recover lead from the bacteria. However, Dr. Swathi recommended using agents like HNO3 to precipitate lead, as heat might denature the protein and make the system unsustainable. We followed her advice in our wet lab and received positive results.



Meeting Recordings


Dr. Arati Ramesh:
Link to meeting recording



Dr. Krishna Ravi Srinivas:
Link to meeting recording



Prof. Sanjay Ghosh:
Link to meeting recording



Dr. Shyam Pingle:
Link to meeting recording



Prof. Srikanth Mutnuri:
Link to meeting recording



Dr. Swathi Alagesan:
Link to meeting recording