Co-Lab Bioremediation

CO-LAB BIOREMEDIATION

26-28 Nov 1

Copyright Š 2017 All rights reserved. This book or any portion thereof may not be reproduced or used in any manner whatsoever without the express written permission of the publisher except for the use of brief quotations in a book review.

Printed in the United Kingdom First Printing, 2017 University College London Gower Street WC1E 6BT www.openscienceschool.org/co-lab/

CO-LAB INTERDISCIPLINARY WORKSHOP

BIOREMEDIATION 26-28 NOVEMBER 2016 DESIGN DEVELOPMENT + SCIENTIFIC EXPERIMENTS + PROTOTYPING HOSTED BY: DEPARTMENT OF BIOCHEMICAL ENGINEERING INSTITUTE OF MAKING UNIVERSITY COLLEGE LONDON

This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 709443.

3

MISSION Our mission is to identify and clean up the poorest communities throughout the developing world where high concentrations of toxins have devastating health effects. Pure Earth devises clean-up strategies, empowers local champions and secures support from national and international partnerships.

Cleaning up one community at a time brings us closer to a Pure Earth.

IMPACT We have cleaned up toxins in 80 locations that were affecting over 4 million people — 20% being children under six. These people will live longer, have less intellectual impairment, cancer and other diseases.

5

EXECUTIVE SUMMARY This report presents activities in the Co-lab Bioremediation Workshop held from 26 - 28 November 2016. The workshop was organised under the collaborative efforts of the Department of Biochemical Engineering at University College London and Pure Earth (pages 38 - 41) and Open Science School. It highlights the programme (page 35) and the appoach adopted during the 3 day workshop. The report outlines Co-Lab’s manifesto (page 17), philosophy (pages 21 - 23) and methodology (pages 26 - 35) concluding with the projects created by the participants at the end of the workshop (Section 4, pages 86 - 111). The report further presents an observation-based critic, reviewing different stages of the workshop - re-orienting itself in a manner so as to evolve and improve on the good practices (Section 5) and ethics adopted during the intensive workshops.

Bioremediation is the use of plants and microorganisms to remove or sequester pollutants. The programme attempts to tackle, discuss topics from a broad perspective ranging from the intrinsic laboratory - based research to its respective extrinsic deployment. (Section 1).

The workshop began with an Introduction by Pure Earth, wherein, they presented the increasing criticality of current toxic metal contamination of water bodies across the globe and its sever effect on the human population (page 38 - 41). This was followed by an empathy workshop allowing the participants to understand the mindset of the diverse group of stakeholders involved in the global environmental crisis (page 42 - 43). The momentum of the workshop exponentially developed, with experiments conducted in the laboratory (Section 3, pages 52 - 85) and interactive lectures - both, generic and technical informing the participants the state of the art of Bioremediation at UCL (pages 78 - 83). The final stage of the workshop inspired the participants into a brainstorming activity exploring design thinking (Section 4, pages 86 - 95). Day 3 of the workshop commenced at the Institute of Making where participants worked on developing their projects / prototypes(pages 98 - 111). The report concludes by highlighting good practices that can be increasingly adopted communities interested in organising workshops like the Co-lab (pages 112 - 117).

7

IDENTIFYING & UNDERSTANDING THE PROBLEM

CO-LAB

BAL-OC

NOITAIDEMER OIB

BIO REMEDIATION

SECTION TWO

SECTION ONE PHILOSOPHY OF CO-LAB

DESIGNING WITH NATURE SECTION FOUR

SECTION THREE UNDERSTANDING THE SCIENCE

SECTION FIVE REFLECTION ON WORKSHOP DESIGN / FORMAT 9

TABLE OF CONTENTS Acknowledgments PART I Manifesto

Introduction Philosophy

Content of the workshop

Structure of workshop Schedule PART II: IDENTIFYING AND UNDERSTANDING THE PROBLEM Understanding the Stakeholders Blacksmith Institute, Pure earth, India Current Situation in India Case Study - Coca Cola Bottling Plant Growing Empathy Biogeochemical Cycles: The Chemical Flows in Nature

11

TABLE OF CONTENTS PART III: UNDERSTANDING SCIENCE Understanding Cadmium(Cd): The Heavy metal ENRICHMENT OF HEAVY METAL TOLERANT MICROORGANISMS Objective Lab Practical Materials Protocol Results CUSTOMISABLE ULTRE-FLTRATION MEMBRANES FROM BACTERIAL CELLULOSE (MADE FROM KOMBUCHA) What is Kombucha What is Bacterial Cellulose Objective Lab Practical Protocol Materials ALGINATE ENCAPSULATION OF Chlorella sp.

Understanding Bioencapsulation Lab Practical Materials Protocol Troubleshooting CADMIUM(Cd) UPTAKE EXPERIMENTS What are Microalgae ?

Microalgae for Bioremediation Objective Lab Practical Protocol Results BIOLOGY 101: PEER-TO-PEER LEARNING STATE OF THE ART BIOREMEDIATION Introduction Bioremediation Mechanisms Defining criteria for Bioremediation Technologies Bioremediation with Microbes

TABLE OF CONTENTS PART IV: DESIGNING WITH NATURE ECO-CENTRISM: NATURE - CENTRED DESIGN CK (CONCEPT - KNOWLEDGE) THEORY Induction @ Institute of making, UCL DRIVING CREATIVITY WITH SCIENCE Project 1: Self Irrigate / Low-tech Bioreactor Abstract Detailed Description Scientific References Project 2: Bio Bucket Chromium Bioremediation Abstract Detailed Description Scientific References Project 3: Chrom-action! Replacing Chemical ETPs with Biological ETPs Abstract Detailed Description Scientific References Project 4: Citizen-Lead (Pb) Detection Abstract Detailed Description Scientific References Project 5: Fungi Edu Kit Abstract Detailed Description Scientific References PART V: CONCLUSION Learning Outcomes and Good Practice from Organisers Conclusion, References, Future Perspectives

13

ACKNOWLEDGMENT We express our deepest appreciation to the ever-growing team of enthusiasts who provided complete support and guidance, in developing and executing the workshop to the best of it’s abilities, exploring to implement a format of teaching - learning, wherein, the idea of interdisciplinarity lies at the core.

We offer special gratitude to our Project/Research Leader, Dr. Brenda Parker, without whose guidance and support this workshop wouldn’t have been possible. We are ever grateful to the ambitious energy you impart to the spirit of CoLab since its commencement in 2015.

Furthermore, we would also like to acknowledge with much appreciation the crucial role of Dr. Laura Stoffels, for all her contributions and preparation of the materials for the Department of Biochemical Engineering and Institute of Making who gave the permission to use all required facilities to host the workshop.

We express our special acknowledgement to Promila Sharma Malik and David Hanrahan from Pure Earth and Reema Banerjee from the Centre for Environmental Education, India for joining our 3-day workshop, and bringing with them a plethora of expert knowledge and experience - from which the participants and the organisers have collectively benefitted. We recognize and thank our team members from Open Science School for their time and effort in bringing together the programme and material for the workshop.

We thank ESPRC Global Challenges Research for the funding of our project. We acknowledge that this project has been carried out within the Doing It Together Science Project (DITOs). This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 709443. special thank you to the DITOs management team at CRI/ Paris Descartes University: Amodsen Chotia (DITOs Coordinator), Imane Baïz (DITOs Project Manager), Juanma Manuel Garcia Arcos and Lena Asai.

University College London, Gower St, Kings Cross, London WC1E 6BT, UK Lena Asai, Juanma Garcia and Shneel Malik

15

SECTION ONE

MANIFESTO We believe that Sciences and the Arts are both creative fields of study. And neither can do without the other.

Our workshop is a place where artists, designers, and scientists meet to initiate collaboration. We aggregate artists and designers to learn biology. We encourage scientists to value and learn artistic approach and design thinking. We bring artists, designers, and scientists together to explore the possibilities of biological design.

The goal of the Co-lab workshops project is to foster the creation of truly interdisciplinary projects around life sciences.

Interdisciplinarity is a tool to solve complex problems that are beyond the reach of any discipline alone. To be able to do this, many soft-skills need to be developed.

However, we believe that they are central to face the challenges of this emerging new world: conceptualization, inter-cultural communication, project-based learning, adaptation, and willingness to learn.

We also believe that 1 + 1 does not equals 2, but sums much more. Being able to exchange knowledge is the most valuable tool that a community can have. Being able to use the skills that we have learned from our field in another discipline makes us more learned and valuable individuals.

17

SECTION ONE

INTRODUCTION Bioremediation is the use of plants and microorganisms to remove or sequester pollutants. Collaborating with partners from the Blacksmith Institute based in India, we expect to generate projects on deployment of technology, as well as explorations on how to create new value chains and new job prospects related to bioremediation. #LabToRealProblems #collaboration #pollution #biocolab #Bioremediation

The workshop consisted of 3 days of activities that bridges together local artists, scientists and designers to brainstorm and work on an interdisciplinary project around bioremediation. Inviting Promila, Pure Earth and Reema from CEE, India to the UK was our effort to promote an alternate effort to discuss pollution - how can people from opposite sides of the spectrum - scientists and citizens discuss the cutting edge research and reality of heavy metal contamination in India? More specifically, it will bring scientists the opportunity to learn about design thinking and ethnographic methodology in a science. Designers will have the opportunity to gain exposure to the lab and interact with local scientists.

This workshop has been organized by: Open Science School, the Department of Biochemical Engineering at UCL, the Blacksmith Institute.

This workshop is has been organised by: Open Science School, Doing It Together Science at Centre for Research and Interdisciplinarity (CRI), The Department of Biochemical Engineering at UCL and the Blacksmith Institute.

We thank ESPRC Global Challenges Research for the funding of our project. We acknowledge that this project has been carried out within the Doing It Together Science Project. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 709443.

19

SECTION ONE

CO-LAB PHILOSOPHY There is a relatively clear reason why the design, arts and science should meet they are all creative disciplines. Arts, design, and science all have the capability of changing the ways humanity views the world.

Collaboration between these disciplines have extensively taken place in the past.

More specifically, the organisers of the workshop were united under the question of the meaning of true interdisciplinarity.

We initially asked ourselves “how do we facilitate the communication of science and design?� We were inspired by the Bauhaus teaching method which replaced the traditional pupil-teacher relationship with the idea of a community of artists working together. Learning by teaching allows a very effective transfer of knowledge, because it makes participants stronger in the skills that they master and also creates a space to acquire new skills. In order for true collaborations to happen, mutual respect must exist. These dynamics flip the teacher-student role constantly and effectively create an horizontal scenario for interaction to happen.

We challenge the gap existing between these disciplines creating a universal language for interdisciplinary idea generation. During the Bauhaus Weimar period, the workshops were co-led by a craftsman as a master of works and an artist as a master of form. The intention was to absorb the arts into the crafts, but a differential focus created a strong imbalance towards the mass production, and after 1923 this approach widely changed. We also co-assign the leading role of the courses to a scientist and a designer and show how real conversations, controversies and agreements take place between them, so that the participants make up their own perspective.

21

SECTION ONE

CO-LAB PHILOSOPHY “Scientists are very much entangled in their culture and this culture is not pristine, untouched by other cultures and practices.” - Bruno Latour Bruno Latour’s theory of construction of scientific facts explains how science lacks contribution from other disciplines. We reverse Latour’s anthropological study in laboratories to actually involve citizens in the brainstorming for scientific projects. We believe that collaboration should be pursued from the point of idea generation, and not just once the technology is released. Co-Lab is our attempt to prove that such an effort is needed to connect science with people from different disciplines to challenge ways synthetic biology can be utilised in various settings and scenarios.

Through the collaboration and discussions in the workshop, we hope the participants will be inspired to bring back these workshops to make it happen in their city and spreading the significance of interdisciplinarity. Most importantly we hope these workshops will encourage participants to connect and form valuable relationships and help to bridge the gap between various disciplines. At the end we believe we need to start creating a community of people who are interested in discussing and engaging with these issues larger than what a single discipline can manage.

Technology creates desirable solutions for growing demands of our human population. However, human beings do not always desire efficiency-driven solutions.

The reality of human nature makes it challenging to introduce new values, rituals and customs into cultures. For example, our world invested a substantial amount of resources to develop genetically modified crops as a food replacement, but a large sector of the public views them with skepticism. This introduces an interesting idea of how the narrative actually transforms the trajectories of technology and progresses them into never imagined territories.

If scientific research has a clear awareness of the needs, values and relationships to be nurtured with relationship to the world from its very conception, the blood and sweat potentially wasted on it could be avoided. When ethical values, religion, philosophy, and culture of the world conflicts with technology, it is virtually impossible to overturn public perception. This becomes harder once the technology is labeled as being controversial. The complexity of this wicked problem requires divergent perspectives, from science, social science and design.

23

SECTION ONE

Collaborating with young, passionate minds to inspire the development of a ‘technologically-sound’ and ‘ethically-advanced’ mature future, that respects the importance of ‘fostering’ what we have around us, while we develop a world of our own.

25

SECTION ONE

INSPIRATION DESIGN SCENARIO - CHANGING TIMES The fields of design and Innovation, are facing a pardigm shift from the otherwise ‘static’ and ‘robust’ non-responsive systems into components designed using synthetically composed ‘technological mechanisms’ to generate prototypes that are increasingly ‘resilient’ and ‘responsive’.

In recent times, efforts are being made by Scientists, Researchers, Engineers, Designers, Innovators alike to understand and define a future that is biologically integrated within the ‘creative’ practice. We are beginning to increasingly speculate the future of design, however, what majorly lacks is the true physical realization / means of achieving such ‘responsive’, truly-productive, ‘active’ - technology.

The thought of amalgamating design concepts with advanced researches within the field of sciences - such as biochemical engineering, synthetic biology, tissue regeneration, biomedicine, etc. - are as new as the changing times itself. Therefore, it is increasingly becoming necessary for such researches to progressively take place on a large-scale to ultimately, bring to reality a way to achieve a longterm growth of the speculated ‘utility - based’ design scenario.

With this comes a major shift in design thinking with labs and studio practices merging to create new sorts of prototypes to experiment with new biological materials, forms, processes. This workshop is an active initiation into exploring a new way of considering the relationship between diverse disciplines, such as synthetic biology, engineering and design, allowing ‘designing’ to move away from the idiosyncratic formal expression and towards ‘designs’ that are increasingly biologically responsive and adaptive to the vicissitudes of its local environment.

Mental and Emotional technology comes from the participants. Everyone’s approach and skills are different and this is what the structure of the workshop increasingly encourages and uses as a positive driving factor, underlying the principles of the Co-Lab.

27

SECTION ONE

INTERDISCIPLINARY APPROACH TOWARDS ‘INTELLIGENT DESIGNING’

29

SECTION ONE

APPROACH DESIGNING THE STRUCTURE OF THE WORKSHOP AIMS 1. To initiate an integrated ‘Innovation’ Culture between experts of different fields - such as, Designers, Architects, Engineers, Scientists, Computer Experts, Artists.

2. To understand and to find ways of dissolving boundaries between disciplines, in a manner so that each benefits from the oether.

3. To introduce a cross-linked learning process, wherein:

- Designers are introduced to the Scientific Research ‘Lab’ Culture

- Scientists are introduced to the ‘Prototype-based’ design development

Creative Culture; This aims to build a progressive understanding and find common grounds of ‘Creative Innovation’, in the emerging fields of ‘Interdisciplinary Research’.

METHOD 1. We identify a certain subject / issue - in this case, ‘Bioremediation’ and design the entire workshop around achieving ‘research-based’ design solutions;

- that are inspired from Scientific Researches

- while, accomodating productive designing ‘principles’ This collaborative effort, aims to encourage the development of a new ‘Innovations Lab’ - that runs through the collborative cooperation of researchers from a vast range of fields.

31

SECTION ONE

APPROACH This 3-Day Intensive Workshop aims to answer the folliwng focus - questions: 1. What is Bioremediation ?

2. Understanding the current scenario of water pollution - its causes and effects on the :

- Humans, and the

- Environment 3. What are the traditional scientific bioremediation techniques ?

How do they work, experimentally and technically ?

4. How can we make them available to the user on a larger scale ?

5. In which way can Design contribute in executing the inclusion of such scientific research-based systems into the daily lives of people.

6. Where are the most appropriate control/nodal points, for introducing ‘designed-scientific’ systems wherein, the technology and the prototype would make maximum impact.

7. How will the stakeholders benefit from this approach ?

8. In what way can this design-technology be multiplicated and utilised int he bioremediation of other toxic metals besides Cadmium (Cd).

9. What are the most suitable / rather what are the varied principles of design that one learns / or thinks is appropriate for researchers to learn / understand while constructing or developing their scientific research, simultaneously, To what extent can the designers be involved in the developmental process of ‘scientific research’ so as to expand the limitations of laboratory - research, allowing them to pro-actively exploit their technology in making it available at a largescale in a manner most suitable for a growing mass consumption.

33

SECTION ONE

SCHEDULE

35

IDENTIFYING & UNDERSTANDING THE PROBLEM SECTION TWO

Toxic Environmental Contaminants pollute Large Water Reservoirs instigating Serious Health Problems World Wide

37

References http://www.blacksmithinstitute.org

SECTION TWO

BLACKSMITH INSTITUTE, PURE EARTH, INDIA INTRODUCTION Toxic pollution poses health risks to over 200 million people, particularly children, in low- and middle-income countries. This is a finite problem that can be solved in our lifetime.

Blacksmith Institute works in some of the world’s worst polluted places, sharing resources and expertise with local groups and agencies to solve pollution problems, clean up polluted sites, and save lives.

References: http://www.blacksmithinstitute.org

CURRENT SITUATION IN INDIA DEMOGRAPHICS - CADMIUM (Cd) POLLUTION Out of 1934 water samples, 7 samples were found to have cadmium content more than the acceptable limits. BIS (Bureau of Indian Standard), 10500:2012 have recommended an acceptable limit of 3 μg/L of cadmium in drinking water.

Total four Indian Rivers viz. Cauvery, Pennar, Yamuna and Hindon are contaminated through cadmium at 7 water quality monitoring stations.

The highest cadmium concentration (4.0μg/L) was observed in the Delhi Rly Bridge and Mathura water quality monitoring station at Yamuna River during June, 2012.

References: Central Water Commission Ministry of Water Resources Govenment of India

39

PLACHIMADA, KERALA, INDIA

LOCATION

KERALA, INDIA

REGION

SOUTH ASIA

POLLUTANTS

LEAD, CADMIUM

SOURCE

GENERAL INDUSTRY

TRANSMISSION

WATER, SOIL

POTENTIALLY AFFECTED PEOPLE

4,000

SECTION TWO

CASE STUDY COCA-COLA BOTTLING PLANT The Problem The Coca-Cola bottling plant at Plachimada is located along the Palakkad-Meenakshipuram-Pollchi road, around three kilometers to the north of the Meenkara dam reservoir and a few hundred meters west of the Kambalathara and Vengalakkayam storage reservoirs.

The bottling plant started production in 1998 on a 42- acre plot in violation of the Kerala Land Utilisation Act, 1967, intended to prevent the use of agricultural land for non-agricultural purposes. Hindustan Coca-Cola Beverages Limited (HCBL) has faced a host of complaints and agitation from local people over water and soil pollution. The issue has been raised in the media with a focus on depletion of water and its contamination. Test results of the well water and the sludge have proved the presence of contamination.

Health Impact Cadmium is a carcinogen and can accumulate in the kidneys with repeated exposure, possibly causing kidney failure.

Ingestion of lead can cause chronic lead poisoning, anemia in children, abdominal pain, lead encephalopathy and paralysis.

Diseases with high relative risks in the adjacent villages are hair-loss, burning of eyes, cough, vomiting, pain in limbs etc. Hair loss is 13 times higher and burning in the eye 7 times higher in the adjacent village.

Current Activity The local Coca Cola distributor has taken these issues very seriously, and has initiated a detailed program to stop further contamination and depletion of the water supply. This work is being overseen by Coca Cola’s head offices. There is no expectation of future problems here.

References http://www.blacksmithinstitute.org/projects/display/141

41

Design Credit: Lena Asai

SECTION TWO

GROWING EMPATHY UNDERSTANDING THE DIFFERENT STAKEHOLDERS IN BIOREMEDIATION SCENARIOS BY ADOPTING ROLE PLAYING Various issues are explored utilising the empathy map, encouraging a growing discussion evolving the participants to think and understand the stakeholders perspective.

Co-lab is not just about understanding visual thinking strategies while being a biologist, neither just about learning how carbon “flows” in nature while being an architect. What co-lab is mostly about, is understanding how people from different disciplines think, rather than what they think. With regards to this, no collaboration can be achieved to any level -from a small interdisciplinary team to a policy maker level- unless stable communication and comprehension bridges are established first. The soft skill that enables us to build these communication bridges, is called empathy! And regardless the big debate on whether empathy is an innate or an acquired skill, everyone agrees that there are some “universal” means of developing empathy; storytelling, role-playing and recently even virtual reality are only a few among them.

Under this prism, during our co-lab we used dialogue and one-to-one debate between different stakeholders, thus, trying to understand all the different viewpoints around our case study in India and the possible ultimate application of bioremediation on it. Proposing alternative solutions seems to be a more effective strategy than raising restrictions and barriers among different stakeholder groups. Common ground can be reached through communication and mutual understanding, open dialogue and interaction. And this is something we achieved during our stakeholder’s debate. Prior to that, the different stakeholders facilitated the understanding of their own points of view and arguments by mapping them with the help of the Empathy Map (see picture...). Thus, after building the understanding of oneself, the one-to-one dialogues began; arguments were contradicted and exchanged and although no common consensus was reached, the understanding between the different stakeholders raised significantly, according to their testimonials once the debate was over.

43

Billions of tons of carbon per year. Yellow numbers are natural fluxes, red are human contributions in billions of tons of carbon per year. White numbers indicate stored carbon.

Image Source: http://www.geo.hunter.cuny.edu/tbw/soils.veg.fall.2016/lecture.outlines/ecology.chap.23/chapter_23.htm

SECTION TWO

WHAT ARE BIOGEOCHEMICAL CYCLES: THE CHEMICAL FLOWS IN NATURE INTRODUCTION TO BIOGEOCHEMICAL CYCLES AND ITS RELATION WITH BIOREMEDIATION Have you ever wondered why the expression ‘of the dust we come and in powder we will become’?

If biogeochemistry was a person, for sure she/he could explain this in detail. But so far we have to content ourselves with the existent literature, that defines biogeochemistry as the scientific discipline that involves the study of chemical, physical, geological and biological processes that govern the composition of the natural environment.

In particular, it deals with control of the concentrations and cycling of chemical elements such as Carbon, Nitrogen, Sulfur, Phosphorus, Hydrogen, Oxygen. As learned by the Law of Conservation of Matter, atoms cannot be destroyed or created, instead they recycle themselves, so these cycles show how the different types of atoms are transformed and used by consumption.

Scientists working in biogeochemistry aim to study the most fundamental aspects of knowledge of the planet Earth and its natural cycles, but in the same way, they also seek a better understanding of extremely important practical problems; most notable among such problems are those caused by human manipulation in biogeochemical cycles on nature’s own scale. Anthropocene defines Earth’s most recent geologic time period as being human-influences based on overwhelming global evidence that all the earth systems processes mentioned before are now altered by humans.

Humans are one of the largest consumers on Earth and hands down use and impact more of the world’s supply of natural resources than any other species, mostly due to overconsumption. All the biogeochemical cycles are actually impacted daily. Some of the most talked about effects have to do with Carbon, Phosphorus and Nitrogen cycles.

45

involvement.

Below, there is a sample of the gradual evolution of our presentation. Credits to Alicia Mansilla Sanchez for all the drawings, as well as to James Carlson for his valuable contribution in using the Prezi software effectively.

Step 1.​ First general picture, presenting the different elements, as well as the different Figure 1: First General Picture, presenting the different elements, as well as the different three spheres of Earth between biogeochemical mainlywhich take place. threewhich spheres of earth cycles between biogeochemical cycles mainly take place.

16

Figure 2: Zooming into a more representative version of the Water Molecule - The Molecule of Life.

Step 2. ​Zooming in a simplified representation of the water molecule.

SECTION TWO

Humans are cutting down forests for an ever-growing population which is lessening the amount of CO2 transferred to oxygen, lowering our air quality. To make matters worse, over consumption of fossil fuels are putting large amounts of carbon into the atmosphere, causing a loss of a natural protector from the sun, the ozone layer, and contributing to a massive greenhouse effect and thus to the acceleration of climate change. In the case of Phosphorus, an important mineral nutrient needed in all the ecosystems. Humans are affecting this cycle with the overuse of fertilizers rich in it, that are finding their way into the ocean, where the excess is wreaking havoc on the ecosystems. With most phosphorus getting trapped essentially in the ocean, ocean life becomes tainted with over fertilization that can lead to death of plant and animal life. In the case of nitrogen, that is also needed by most organic material to survive, humans impact its cycle dramatically through industrialization, resulting in more than double the natural amount of nitrogen from the atmosphere into the soil. Excess nitrogen is finding its way into the soil, where it sucks surrounding land dry of minerals that lead to mineral deficiencies.

Moreover, it is also becoming in excess in the water supply, seriously affecting plant and animal lives. And these are not the only actors on scene. Actually, the pollutants that are of environmental concerns come from the combination of these chemical elements, among others, in the form of toxic compounds (e.g hydrocarbons, greenhouse gases, pesticides, heavy metals and nuclear wastes). Given this scenario, action must be taken. And here is where bioremediation comes on scene. Remediation of polluted sites using microbial process (bioremediation) has proven effective and reliable due to its eco-friendly features. Choosing appropriate bioremediation technique can effectively reduce pollutant concentrations to an innocuous state. For example, a wide range of microorganisms, including bacteria, fungi, yeasts and algae, can act as biologically active detoxifiers of toxic species. Although these microorganisms cannot destroy metals, they can alter their chemical properties via a surprising array of mechanisms. Also some microalgae could serve as ‘fertilizer-cleaners’ in eutrophicated waters, given the fact that they nourish from the macro and micronutrients contained in them.

The aim of this lecture is to familiarize participants with the main biogeochemical cycles, to make them aware of how do we humans daily disrupt some of the most fundamental natural processes and to show to which extent is it also in our hands and under our responsibility to remediate this human-driven situation; also how bioremediation is enormously helpful in this context and some practical examples.

47

Step 2. ​Zooming in a simplified representation of the water molecule.

Step Zooming even on the earthcycles and -taking a firstthe look toCycle the incycle of water.

Figure 33.​:Looking closely at themore Planet’s geochemical understanding Water the Atmosphere.

17

Figure 4 :Step-by-Step explanation of the Water Cycle - one of the most dominant biogeochemical cycles on the Planet Step 4.​ Zooming in even more to start the step by step explanation of the cycle of

water.

Understanding the Science:

SECTION TWO

WHAT ARE BIOGEOCHEMICAL CYCLES: THE CHEMICAL FLOWS IN NATURE DESIGNING THE PRESENTATION FORMAT AND PEDAGOGICAL OBJECTIVES To discuss also a little bit about the format chosen for the biogeochemistry lecture is worth mentioning that one of the unique points about a co-lab, is that you do not necessarily need to be an expert in order to facilitate a concept presentation, as long as you find the effective way to do it. In our case, the approach to the biogeochemical cycles was not that difficult -given our background in life sciences.

However, while preparing for this co-lab, we needed to run again through some theory and most importantly, we had to find the best possible way to transform this theory into a format easily comprehensible from a diverse audience; our audience varied from biochemists and engineers to designers and architects.

We, thus, brainstormed around the playful -yet meaningful- format that our presentation on the biogeochemical cycles could have. Eventually, we used an innovative, online presentation software, Prezi; this software provides with the unique trait of gradual transition from one point to the other, thus, scaling up the difficulty of the concepts explained and making clear steps from one concept to the other.

Most importantly, though, the use of this presentation software allows to finally bring all the different pieces of knowledge together in one “big picture”, thus, facilitating learning and memorizing. Moreover, instead of using google images to explain the concepts, we drawed by hand the different biogeochemical cycles in order to give a more playful tone to the presentation, and thus, make it more appealing and facilitate the audience’s involvement.

Left, there is a sample of the gradual evolution of our presentation. Credits to Alicia Mansilla Sanchez for all the drawings, as well as to James Carlson for his valuable contribution in using the Prezi software effectively.

49

SECTION TWO

WHAT ARE BIOGEOCHEMICAL CYCLES: THE CHEMICAL FLOWS IN NATURE Conclusions, Reflections and Future Perspectives As we already mentioned above, one of the greater challenges addressed in a co-lab is the “common language” that is needed in order to efficiently communicate diverse scientific concepts to people from diverse backgrounds. Different concepts (ranging from biochemistry to design thinking in our case) need to be essentially understood in little time, as the participants are later asked to work on the interface between this concepts.

However, how can we evaluate the depth in which co-lab participants understand new concepts after presentations?

One proposal for future co-labs could be to provide participants with questionnaires so that they can evaluate themselves on the new knowledge they gained during a presentation. Moreover, innovative questionnaire approaches could be adopted (e.g. a gamified questionnaire that links all the different concepts presented).

Co-lab is an innovative approach to knowledge-sharing and interdisciplinary science and collaboration. Thus, their format is dynamic and constantly changing and evolving. What could we do to escape from the typical presentation format?

How could we introduce innovative pedagogy tools to facilitate even more participants’ learning and expand the innovative setting of co-labs?

51

UNDERSTANDING SCIENCE SECTION THREE

Participants of the Workshop, getting familiar with the Lab Protocols before beginning to perform Experiments

53

David Hanrahan from Pure Earth is seen here interacting with the Participants of the Workshop, getting Hands - on with the Experiments

SECTION THREE

UNDERSTANDING SCIENCE The Co-lab workshop about bioremediation included four practical laboratory sessions. The aim of these sessions was to include, between the lectures and design workshops, some hands on experiences for all participants, and to give the non-scientists the chance to work in a laboratory.

For the bioremediation Co-lab all laboratory sessions were designed to demonstrate to the participants that some biological organisms can adapt to pollutants such as heavy metals, can bind or take them up and decrease their toxicity. The purpose of the sessions was to show these processes to the participants first hand and to give them examples of biological materials that could be used for bioremediation projects.

55

UNDERSTANDING CADMIUM (Cd) - THE HEAVY METAL Open Science School’s Co-lab Bioremediation

Sources of Cadmium Pollution in Water References http://www.mdpi.com/sensors/sensors-11-10638/article_deploy/html/images/sensors-11-10638f8-1024.png

Content: What is cadmium? When is it released to nature? How is cadmium toxic?

Resistance of microorganisms to heavy metals.

Activities: Isolation of microorganisms from the environment with a high tolerance to cadmium

THEORY SECTION THREE

INTRODUCTION What is cadmium?

Cadmium is a metal found in the earth's crust, associated with zinc, lead, and copper ores.

It is a chemical element with the symbol Cd and atomic number 48. Pure cadmium is a soft, silver-white metal. Cadmium chloride and cadmium sulfate are soluble in water. It is used in the production of certain batteries (mainly rechargeable nickel-cadmium batteries), pigments, coatings and platings, stabilizers for plastics, nonferrous alloys and photovoltaic devices.

Cadmium is often extracted as a byproduct during the production of other metals such as zinc, lead, or copper. Cadmium is emitted to soil, water, and air by non-ferrous metal mining and refining, manufacture and application of phosphate fertilizers, fossil fuel combustion, and waste incineration and disposal (e.g. of batteries). Cadmium and its compounds may travel through soil, but its mobility depends on several factors such as pH and amount of organic matter, which will vary depending on the local environment. Generally, cadmium binds strongly to organic matter where it will be immobile in soil and be taken up by plants, eventually, entering the food supply. Elevated cadmium levels in water sources in the vicinity of cadmium emitting industries (historical and current) have been reported. Aquatic organisms will accumulate cadmium and possibly entering the food supply. Workers can be exposed to cadmium in air from the smelting and refining of metals, or from the air in plants that make cadmium products such as batteries, coatings, or plastics. Workers can also be exposed when soldering or welding metal that contains cadmium.

TOXICITY Cadmium acts as a catalyst in forming reactive oxygen species. It increases lipid peroxidation and additionally depletes antioxidants, glutathione and protein-bound sulfhydryl groups.

It also promotes the production of inflammatory cytokines. Severe exposures to cadmium fumes can cause tracheo-bronchitis, pneumonitis, and pulmonary edema. Inhaling cadmium-laden dust quickly leads to respiratory tract and kidney problems which can be fatal (often from renal failure). Ingestion of any significant amount of cadmium causes immediate poisoning and damage to the liver and the kidneys. Compounds containing cadmium are also carcinogenic.

Resistance of microorganisms to heavy metals Cadmium and other heavy metals are part of the chemical elements and have been present since the beginning of the earth. Some microorganisms evolved therefore mechanisms to survive in the presence of increased heavy metal concentrations. Several different resistance mechanisms have evolved, including efflux and sequestration mechanisms. Some cations for example can be pumped out of the cell or sequestered by chelates like thiol-containing compounds, while others can be reduced to a less detrimental state. Bacteria that are able to form biofilms are often more resistant to heavy metals as well as antibiotics.

References Agency for Toxic Substances and Disease Registry of the US government (https://www.atsdr.cdc.gov/). www.wikipedia.org MicrobeWiki (https://microbewiki.kenyon.edu/index.php/MicrobeWiki) Hassen et al (1998) Resistance of enviromental bacteria to heavy metals. Bioresource Technology 64.

57

Enrichment of heavy metal tolerant microorganisms Open Science School’s Co-lab Bioremediation

On - Site photograph of a Heavy Cadmium contaminated Site References http://www.newslodi.com/wp-content/uploads/2016/05/pollution.jpg

OBJECTIVE Living organisms such as animals, plants and microorganismscan are adapted to their individual environment and thrive in their biological niche. There are also organisms that adapted to pollutants that would kill most other organisms. These organisms have evolved strategies to cope with the toxic pollutants or to decrease their toxicity. Therefore, in some cases these organisms can be employed to remove or detoxify pollutants such as heavy metals and are of interest for bioremediation projects.

In this experiment, the participants collected samples from places around UCL. They were encouraged to choose places where they suspected to find cadmium resistant bacteria and to take a picture of the sampling site. Subsequently, they put the samples on nutrient agar plates, which encourage the growth of a range of microorganisms. The plates contained different concentrations of cadmium (0, 200, 400 and 800 mg/L) and were subsequently incubated for 48 h at 30ÂşC. Additionally, some plates were inoculated with soil samples from four places in India that are known to have a high concentration of cadmium and other heavy metals in the soil.

LAB EXPERIMENTS SECTION THREE

Two samples came from landfill sites, one of them from the dump and one from the leachate.

Another one was collected outside of a medical incineration plant and the last one from a battery recycling site. The idea was to compare these samples to the samples collected around UCL to demonstrate that the presence of cadmium reprents a selective pressure for resistant organisms.

After the incubation period, it was analysed how many organisms grew on the different plates. The samples from the polluted sites contained all organisms that could grow on 200 mg/L of cadmium and three contained several organisms that even grew on 800 mg/L (Graph 1 and Table 1). In contrast, only one sample from around UCL contained an organism that grew on 800 mg/L, less than 10% of samples grew on 400 mg/L and 37% on 200 mg/L, whereas 70% of the samples grew on control plates without cadmium (Graph 1).

Therefore, the experiment showed a clear and expected result. The soil samples from the heavy metal polluted sites contained a lot more cadmium resistant bacteria than the samples collected around UCL. The presence of heavy metals in the soil causes a selective pressure for organisms with an increased tolerance against cadmium. A range of organisms were found on the cadmium plates, some filamentous fungi as well as unicellular eukaryotic microorganisms and bacteria. The pictures show a selection of cadmium resistant microorganisms from around UCL and the Indian sampling sites.

LAB PRACTICAL In this practical you will enrich for microorganisms with a high resistance to cadmium from the area around UCL and a heavy-metal polluted site in India.

MATERIALS LB medium: Yeast extract (5 g/L), Tryptone (10g/L), NaCl (10 g/L), with 0, 400, 800 mg/L Cd as CdCl2.

PROTOCOL Warning: Cadmium is toxic! Wear a lab coat, safety glasses and gloves when you are handling cadmium plates. Do not drink or eat in the lab.

1. Take samples or swabs of different things, soil or water that you can find close to UCL. Choose places or things where you think heavy metal resistant microorganisms might be present and take pictures of each sampling site. Back in the lab, inoculate the agar plates (LB) containing cadmium (0, 200, 400, 800 mg/L) with your samples.

2. We will provide soil samples from a heavy metal polluted site in India as a comparison to sites around UCL. 3.

The plates will be incubated overnight at 30ยบC.

4. On the next day, you can look at the colony and cell morphology of the cadmium resistant organisms you isolated. Which samples contain the most heavy metal resistant organisms? We will match pictures of the organisms with the pictures of the sampling site.

59

Enrichment of heavy metal tolerant microorganisms Open Science School’s Co-lab Bioremediation

Bacterial Growth on Plates with Cadmium (Cd)

Bacterial Growth on Plates without Cadmium (Cd)

Soil Samples from Landfill Sites - Leachate & Dump

LAB EXPERIMENTS SECTION THREE

RESULTS

P+A 200 mg/L

Samples Collected around UCL P+A 200 mg/L

PLANTER 800 mg/L

Samples from Indian Sites

PLANTER 800 mg/L

Samples from UCL with Cadmium Resistant Microorganisms

Table 1: Collection sites of the heavy metal polluted soil samples from India. +++/++/+ Microbial growth on LB medium plates containing cadmium after 48h at 30°C. – No growth.

Graph 1: Enrichment experiment for cadmium resistant microorganisms.

Samples were collected around UCL (top) and at heavy metal polluted sites in India (bottom). The graph shows the percentage of samples that showed microbial growth on LB medium plates after incubation at 30°C for 48h.

61

Customisable ultra-filtration membranes from bacterial cellulose (made from Kombucha) Open Science School’s Co-lab Bioremediation

A Close-up of the Dried Kombucha, post processing

Content: 1. Introduction to Kombucha and Bacterial Cellulose 2. Using Bacterial Cellulose for Water Filtration Activities: 1. Lab: Using Bacterial Cellulose for Water Filtration (Example: Nickel) 2. Brainstorm potential bioremediation applications of bacterial cellulose

References https://www.hive76.org/drexel-design-futures-bacterial-cellulose-and-a-world-record-maybe

THEORY SECTION THREE

INTRODUCTION What is kombucha?

Kombucha tea is a traditional health-promoting fermented beverage that exists since several thousand years. It is a variety of fermented, lightly effervescent, sweetened black or green tea drinks that are commonly intended as functional beverages for their supposed health benefits.

Kombucha is produced by fermenting sugared tea using a “symbiotic colony of bacteria and yeast” (SCOBY). Actual contributing microbial populations in SCOBY cultures vary, but the yeast component generally includes Saccharomyces (a probiotic fungus) and other species, and the bacterial component almost always includes Gluconacetobacter xylinus to oxidize yeast-produced alcohols to acetic and other acids.

Kombucha is a symbiotic living material made of a variable composition of different species of bacteria and yeast. Among the most common ones participating in the symbiosis we can find: 1. Bacteria: Acetobacter xylinum, A. xylinoides, A. aceti, A. pasteurianus, Bacterium gluconicum.

2. Yeasts: Schizosaccharomyces pombe, Kloeckera apiculata, Saccharomycodes ludwigii, Saccharomyces cerevisiae, Zygosaccharomyces bailii, Brettanomyces bruxellensis, B. lambicus, B. custersii and Pichia.

The cellulose matrix formed in the culture medium can be used as a bio-cellulose tissue to create clothes, or bio-paper. But it can also be used as a cosmetology for its anti-inflammatory, antioxidant and anticancer properties.

In natural habitats, the majority of bacteria synthesize extracellular polysaccharides, such as cellulose, which form protective envelopes around the cells. Bacterial cellulose is an organic compound with the formula (C6H10O5)n produced by certain types of bacteria. While cellulose is a basic structural material of most plants, it is also produced by bacteria, like Acetobacter, in kombucha. Bacterial, or microbial, cellulose has different properties from plant cellulose and is characterized by high purity, strength, moldability and increased water holding ability.

Super-wide screen made from a single large sheet of bacterial cellulose “paper”

63

Customisable ultra-filtration membranes from bacterial cellulose (made from Kombucha) Open Science School’s Co-lab Bioremediation

Main: Side view of a BC culture, showing the cellulose pellicle (white “gel” on surface), growth medium and some bacteria/ yeast colonies (dark brown structures). The bubbles are CO2 produced by the yeast.

Inset: SEM micrograph of a bacterial cellulose sample showing a coherent 3-D network formed by cellulose fibers connected by physical joints.

Image Source: http://www.mdpi.com/2079-4983/3/4/864/htm

THEORY SECTION THREE

INTRODUCTION TO BACTERIAL CELLULOSE AND KOMBUCHA A Purer Cellulose...

Cellulose is the most abundant organic polymer (think paper and cotton as examples!) found in nature. However, much of the cellulose used commercially is in impure, as it is derived from plants. Bacteria offer an alternative means of production that produces a cellulose that is purer and so requires less processing for certain applications.

With Many Applications (including bioremediation)…

This so-called ‘Bacterial Cellulose’ (BC) has interesting biotechnological characteristics such as high tensile strength, small pore size and increased water-holding. BC is also a biocompatible material, meaning it does not cause an inflammatory response in the body.

Applications range from biomedical in wound dressings and tissue scaffolds, to high quality papers and diaphragms for high-performance speakers. Finally, the microstructure of bacterial cellulose and its porosity lend itself to applications in bioremediation, especially in water filtration

Using Bacterial Cellulose as an ultrafiltration membrane for Water Filtration Bacterial cellulose naturally adsorbs (holds on its surface) a variety of compounds, in fact this is one theory behind why Gluconacetobacter produce cellulose – for protection. Also, the small pore size of bacterial cellulose means that it is in fact a natural physical barrier to contaminants by acting as an ‘ultrafiltration’ membrane (pore size below 10 nm). Ultrafiltration membranes are able to separate water from a wide range of small contaminants, from bacteria to even some large viruses.

Ultrafiltration is utilised in many fields of water management nowadays, such as wastewater treatment, purification of lab- and cosmetics grade water and drinking water treatment. It is a sustainable method as it does not require the use of chemicals for the treatment and is relatively simple to implement and automate compared to other water treatment methods. Ultrafiltration membranes are also required to withstand large mechanical forces due to the pressure applied to them during the filtration process. Therefore, the material they are made out for should also be strong. Bacterial cellulose is an ideal candidate for this when processed in the right way.

In addition to their natural binding and ultrafiltration properties, BC can be further modified to increase the binding capacity through addition of contaminant-binding proteins fused to a cellulose-binding-domain protein.

This approach was used by a team of students from Imperial College London undertook a project as part of the synthetic biology iGEM (International Genetically Engineered Machine) competition on creating customisable ultrafiltration membranes from BC http://2014.igem. org/Team:Imperial.

65

Customisable ultra-filtration membranes from bacterial cellulose (made from Kombucha) Open Science School’s Co-lab Bioremediation

OBJECTIVE The purpose of the ‘Using Bacterial Cellulose for Water Filtration’ workshop was to explore the potential of how this natural microbial by-product can act as a binder for contaminants in water, using solutions of the heavy metal nickel as an example. In the workshop, participants were given various samples (oven-dried or wet) of bacterial cellulose which had been pre-grown from a kombucha (fermented tea) protocol using a starter ‘SCOBY’ bought from common online retailer. After passing nickel-containing water solutions through the material, participants saw a reduction in amount of nickel compared to the starting concentration, as measured using a quick colourmetric assay read in a spectrophotometer.

The experiments worked successfully, but the slow flow-rate through the material highlighted the importance of design and implementation, and it was great to see many people taking it further to explore different ways to expose the material to the contaminated water which would be more efficient. For example, people tried passing water through several layers, placing lumps straight in the water and incubating for different amounts of time, all of which saw different results and allowed the participants to practise thinking about how to plan their own experiments. All participants enjoyed getting hands-on with the bacterial cellulose (especially the wet samples), and many were amazed that it was produced by microorganisms, and was so easy to grow at home as the by-product of fermented tea!

The workshop used unmodified bacterial cellulose, but the microbial origin of the material lead to several interesting discussions with the demonstrators about how the material’s binding properties can be potentially modified by synthetic biology. It was also very rewarding to see that the material had inspired one group’s final idea in the prototyping workshop on the final day of the CoLab.

Content: 1. Introduction to Kombucha and Bacterial Cellulose 2. Using Bacterial Cellulose for Water Filtration Activities: 1. Lab: Using Bacterial Cellulose for Water Filtration (Example: Nickel) 2. Brainstorm potential bioremediation applications of bacterial cellulose

LAB EXPERIMENTS

LAB PRACTICAL

SECTION THREE

Using Bacterial Cellulose for Water Filtration (Example: Nickel) Details of Bacterial Cellulose Growth Using Kombucha (pre-prepared) The Kombucha protocol we used can be widely found online.

We used the one on the following website, which is based on a protocol from BioCouture: http://2014.igem.org/Team:Imperial/Protocols.

The Bacterial Cellulose was harvested and dried between tissue paper in an oven at 60-70 °C for 16-48 hours. A variety of samples are provided for this workshop: some are still wet and have not been dried. Some thick, some are thinner. You will have chance to touch and play with them all!

1. Filtering Nickel-contaminated water through a bacterial cellulose membrane Note: the Nickel solutions are toxic – ensure you remember to wear your lab coat, gloves and safety glasses.

1) Take a dry BC membrane and cut it to a circle which fits in and covers the bottom of the holder 2) Place the holder over a container to collect the filtrate (e.g. a 50 mL test tube) 3) Slowly apply 2 mL of Nickel Chloride solution (TOXIC) (0.5 mM, or 64.8 mg/L = 64.8 ppm) to the membrane and wait for the water to pass through. If necessary, apply pressure by attaching to the vacuum pump.

4) Test the concentration of Nickel in the filtrate using the Nickel Assay Protocol 2. Nickel assay protocol to measure concentration of Nickel in water solution.

Nickel can form a coloured compound when it reacts with certain sulphur-containing chemicals (thiols). It is this chemistry that we can use to measure the amount of nickel in a solution by comparing the colour change of different concentrations of Nickel.

MATERIALS Nickel Assay Solution: 20 mM NaPO4 (sodium phosphate), 10 mM DTT (dithiothreitol), pH 7.5.

Spectrophotometer: Set to read at wavelength 465 nm.

Cuvettes: Vessel for mixing sample and nickel assay solution and which fits in the spectrophotometer.

PROTOCOL 1) Using a pipette, mix 200 µl of sample to 800 µl Nickel Assay Solution in a cuvette.

2) Incubate at room temperature for 5 min 3) Measure absorbance of cuvette in a spectrophotometer at 465 nm 4) Compare reading to standard curve below to get an estimate of concentration 3. Experimenting with different types of bacterial cellulose In the workshop there are a wide variety of bacterial cellulose samples. Now that you have completed and initial test filtration, experiment with the different types and find interesting ways it could work and which works best. A few examples to get you started: · Several layers of bacterial cellulose? · Float some pieces in solution of nickel, measure concentration after periods of time to see if nickel is adsorbing to the cellulose?

67

Alginate encapsulation of Chlorella sp.

Open Science School’s Co-lab Bioremediation

Chlorella is a genus of single-cell green algae belonging to the phylum Chlorophyta. It is spherical in shape, about 2 to 10 Îźm in diameter, and is without flagella. The cultivation of microalgae in photobioreactors by continuous culture has been used worldwide to generate large amounts of biomass. This type of culture systems is well established and applied to the production of microbial biomass in fermenters (bacteria and fungi), for example, the alcohol industry.

Algae Cultures within the Lab

Content: What is alginate, where do we find it, why do we use it?

Encapsulation at an industrial level.

Activities: Prepare sodium alginate, calcium chloride solution.

Create the alginate beads drop by drop.

THEORY SECTION THREE

INTRODUCTION Alginate is a biopolymer coming from the cell walls of kelp among other algae. Alginate can create mesh-like structures composed of polysaccharides that become insoluble in contact with calcium. They can keep living bacteria, algae, or even enzymes in the inside. They can be immobilized while remaining biologically active or alive. In this way, bioreactors will keep microorganisms inside while water flows constantly.

CONTENT: BIOENCAPSULATION Encapsulation by a permeable membrane could allow for sustained growth without contamination. Porous membrane allows passage of nutrients, but not cells. This can be useful in different settings in biotechnology, like when we are co-culturing two different species together or when we have to isolate biomass from the medium or simply take a soluble molecule and no biomass. The process of separating biomass and medium can be very costly or time-consuming. Aggregating the microorganisms in beads eases this process.

Microencapsulation and alginate, collagen, or cellulose scaffolds for cells have become quite popular recently and have grabbed a lot of attention from the media. Here we want to present a very simple procedure of encapsulation and 3D printing that has already been used for some time in science, and even in kitchen (“cuisine moléculaire”). It was also used to print layer of bacteria in a biofilm-like confirmation by the iGEM team TU Delft in 2015.

The protocol that follows uses encapsulation by calcium chloride. Calcium alginate is a water-insoluble, gelatinous, cream-coloured substance that can be created through the addition of aqueous calcium chloride to aqueous sodium alginate.

Encapsulated Algae through the technique of Immobilisation

69

Alginate encapsulation of Chlorella sp.

Open Science School’s Co-lab Bioremediation

1. Preparing the Algae

2. Suspend algae cells in Sodium Alginate solution

3. Drop by drop, with a syringe drip the algal - alginate solution into a bath of Calcium Chloride

5. Store in cold water

4. Seperate the beads using a sieve Image Courtesy: National Centre for Biotechnology Education

Illustration showing the process of immobilising cultured algae

LAB EXPERIMENTS

LAB PRACTICAL:

SECTION THREE

Encapsulating microalgae MATERIALS Chlorella living culture.

Sodium alginate, Calcium chloride Plastic pasteur pipettes Beakers or plastic cups (not provided) Deionized or distilled water (not provided) PROTOCOL Prepare the following solutions: 150 ml of 4% Sodium alginate solution in water (6 g per 150 ml).

200 ml of 4% Calcium chloride solution in water (8 g per 200 ml).

Chlorella spp. saturated living culture.

Mix the Sodium alginate and the Chlorella solution 1:1.

To create the encapsulated balls or fibers, you just need to mix some concentrated algae solution with the the alginate solution. Drop by drop, pour the alginate-algae mix into the Calcium chloride solution. The alginate-algae drops will become solid when they are floating in the calcium chloride solution.

You can try alternative ways of encapsulating, printing, making wires, or changing the proportions of algae/alginate/calcium.

TROUBLESHOOTING The consistency and stiffness of the resulting objects will depend on you extrusion method as well as the final concentrations of alginate and calcium chloride. The solutions will almost immediately become solid when they are in contact at that concentration (20 grams per litre, or 2% each).

If the beads are too soft, you might want to increase the concentration of each of the components or leave the beads longer in the calcium chloride media. If the beads are too hard, try to add more deionised/distilled water to the alginate-algae solution.

Be careful not to put calcium chloride into the sodium alginate beaker or bottle, because it will polymerize the entire bottle and decrease the efficiency of your experiment.

71

Cadmium Uptake Experiments Open Science School’s Co-lab Bioremediation

Microscopic View of Arthrospira platensis - cynobacterium References http://ccala.butbn.cas.cz/sites/default/files/styles/ccala_big/public/ccala_collection/13256/1352919359-28.jpg

Content: What are microalgae?

Microalgae for bioremediation.

Activities: Following the uptake of heavy metals by algae and the reduction of toxicity Test the toxicity with the bacterium E. coli

THEORY SECTION THREE

INTRODUCTION What are microalgae?

Microalgae are a diverse and polyphyletic group of microorganisms that are capable of performing oxygenic photosynthesis. Some definitions of microalgae are restricted to eukaryotic, photosynthetic microorganisms such as green algae (chlorophyta), red algae (rhodophyta), dinoflagellates (dinoflagellata) and diatoms (bacillariophyceae). However, the term “microalgae” can also refer to prokaryotic, photosynthetic microorganisms and include cyanobacteria (traditionally called blue-green algae). Microalgae can be found in nearly all habitats from marine and freshwater environments to terrestrial habitats and these organisms contribute considerably to the overall carbon fixation on earth.

Arthrospira platensis is a free-floating filamentous cyanobacterium characterized by cylindrical, multicellular trichomes and belongs to the oxygenic photosynthetic bacteria. A dietary supplement is made from A. platensis and A. maxima, known as spirulina. A. platensis occurs naturally in tropical and subtropical lakes with high pH and high concentrations of carbonate and bicarbonate and is mainly found in Africa, Asia, and South America.

Microalgae for bioremediation Biotechnology of microalgae has gained popularity due to the growing need for novel environmental technologies. Inexpensive and sustainable growth requirements (solar light and CO2), and the possibility to use the biomass simultaneously for multiple technologies (e.g. carbon mitigation, biofuel production and bioremediation) make microalgae promising candidates for several eco friendly technologies.

Microalgae have developed an extensive spectrum of mechanisms (extracellular and intracellular) to cope with heavy metal toxicity. There are three general biological processes of the removal of metal ions from a solution by microalgae and other microorganisms: biosorption (adsorption) of metal ions onto the surface of the organism, intracellular uptake of metal ions, and chemical transformation of metal ions by the organism. Biosorption has been reported as the most rapid mechanism. The cell surface of cyanobacteria and microalgae consists of polysaccharides, proteins, and lipids, which can act as a basic binding site of heavy metals. These functional groups within the cell wall provide the amino, carboxylic, sulfydryl, phosphate, and thiol groups that can bind metals. Several studies have shown that alive and dead algal biomass from several species can be used to remove heavy metals from solutions.

References: www.wikipedia.org, Rangsayatorn et al. (2002) Phytoremediation potential of Spirulina (Arthrospira) platensis: biosorption and toxicity studies of cadmium. Environmental Pollution 119 45–53, Kumar et al. (2015) Microalgae – A promising tool for heavy metal remediation. Ecotoxicology and Environmental Safety 113 329–352.

73

Cadmium Uptake experiments Open Science School’s Co-lab Bioremediation

Figure 1: Toxicity of cadmium (final concentration 250 mg/L) on the bacterium Escherichia coli, and the reduction of the toxicity after treatment with the “Spirulina” biomass.

RESULTS: E. coli culture: LB + water: Turbidity = Growth LB + cadmium (500 mg/L) Clear = No growth LB + Cd solution treated with algal biomass: Turbidity = Growth

OBJECTIVE Microalgae, including cyanobacteria, have several charged, functional groups on their cell surface that can bind heavy metals and can be therefore used to bind and remove heavy metals from a solution and potentially the environment.

In this experiment, the participants used dried biomass of the cyanobacterium Arthrospira platensis, commonly known as Spirulina and a popular food supplement, to treat a solution with cadmium. A 20 minutes incubation of the cadmium solution with 5 g/L of the dried algal biomass strongly reduces the concentration of 500 mg/L (Figure 2). To observe the removal of the cadmium from the solution, the participants tested the toxicity of the solution on the bacterium Escherichia coli. Therefore, they mixed the cadmium solution before and after the treatment with an E. coli culture, which was subsequently incubated overnight. The solution with a final concentration of 250 mg/L completely inhibited the growth of E. coli meaning the culture stayed clear. In contrast, the solution after the treatment with algal biomass was not toxic to E. coli any longer and the culture grew, which could be observed as turbidity/cloudiness of the culture medium. A control with water was included to show the participants what the growth of E. coli looks like. Figure 1 shows the result of one group’s experiment. For most groups of participants the experiment worked very well and showed the expected outcome.

LAB EXPERIMENTS

LAB PRACTICAL:

SECTION THREE

In this practical you will use dried biomass of the cyanobacterium Arthrospira platensis to analyse the removal of cadmium from a solution and the following reduction of its toxicity.

PROTOCOL: Warning: Cadmium is toxic! Wear a lab coat, safety glasses and gloves when you are handling cadmium solutions. Do not drink or eat in the lab.

1. Add 4 ml of cadmium solution (500 mg/L) to the freeze-dried “Spirulina” (Arthrospira platensis) biomass (20 mg), pipette up and down, and incubate the mixture on a shaker for 20 min.

2. Prepare three sterile tubes with 2 ml of 2x LB medium. Add 2 ml of cadmium solution (500 mg/L) to one of them and 2 ml of sterile water to another. Both are controls. Label all tubes.

3. Add 50 µl of E. coli culture to all sterile tubes.

4. After the 20 min incubation time, transfer two times 1.3 ml of the spirulina/cadmium mix to two Eppendorf tubes and centrifuge them at 14K rpm for 5 min.

5. Add 2 ml of the supernatant to the remaining sterile tube with 2x LB medium.

6. The tubes will be incubated at 37ºC overnight. Cultures with happily growing E. coli become turbid, inhibited cultures stay clear.

Figure 2: Cadmium uptake by Arthrospira platensis (“Spirulina”) biomass.

A solution with 500 mg/L cadmium was incubated with freeze-dried A. platensis biomass. The figure shows the reduction of the cadmium concentration after 20 min incubation.

Treatment with 5g/L freeze-dried A.platensis biomass

References: Solisio, C. et al. “Cadmium Biosorption On Spirulina Platensis Biomass”. Bioresource Technology 99.13 (2008): 5933-5937. Web.

75

One of the valuable aspects of an interdisciplinary team is the asymmetry of knowledge and skills, which becomes very impactful if managed well among the members. However, when trying to go together deeper on one specific topic, it can be very hard to keep the motivation and the learning curve for everyone. Biologists and people more familiar with life sciences would be very bored if they have to attend a Biology 101 lesson about bioremediation. Biology 101 presented some foundational basis of biology, biochemistry, and bioengineering needed to understand bioremediation and how life produces, stores, and makes energy.

We had a collaborative lecture divided on three different topics: the chemical basis of life and the type of biomolecules; the type of energy metabolisms and diversity of life; and the use of synthetic biology and genetic engineering to change life. The format of the lecture was specifically designed to involve all biologists in the room in the learning and teaching process.

We divided all participants in groups that were composed of a mixture of people already knowing about the subject and people not so familiar with life sciences. We did a few cycles of lecture in the following manner: first, the lecturers were presenting a few topics about biology; secondly, the participants were discussing about this and explaining to each other the main ideas; at the end, we debriefed of the content that we were supposed to learn or follow.

37

THEORY SECTION THREE

BIOLOGY 101 (PEER - TO - PEER LEARNING) THE WORKFORCE OF BIOREMEDIATION One of the valuable aspects of an interdisciplinary team is the asymmetry of knowledge and skills, which becomes very impactful if managed well among the members. However, when trying to go together deeper on one specific topic, it can be very hard to keep the motivation and the learning curve for everyone.

Biologists and people more familiar with life sciences would be very bored if they have to attend a Biology 101 lesson about bioremediation. Biology 101 presented some foundational basis of biology, biochemistry, and bioengineering needed to understand bioremediation and how life produces, stores, and makes energy.

We had a collaborative lecture divided on three different topics: the chemical basis of life and the type of biomolecules; the type of energy metabolisms and diversity of life; and the use of synthetic biology and genetic engineering to change life. The format of the lecture was specifically designed to involve all biologists in the room in the learning and teaching process. We divided all participants in groups that were composed of a mixture of people already knowing about the subject and people not so familiar with life sciences. We did a few cycles of lecture in the following manner: first, the lecturers were presenting a few topics about biology; secondly, the participants were discussing about this and explaining to each other the main ideas; at the end, we debriefed of the content that we were supposed to learn or follow.

77

Belgium, Remediation of the Carcoke site in Brussels

Image Source: http://www.jandenul.com/en/activities/environmental-works/soil-and-groundwater-remediation

Ex-Situ Bioremediation

Image Source: http://www.vertasefli.co.uk/our-solutions/expertise/ex-situ-bioremediation

WHAT DOES BIOREMEDIATION LOOK LIKE? In-situ: Contaminated land or water is treated on site. Physical methods are used to encourage microorganisms e.g. adding a carbon source, sparging air or turning soil.

Ex-situ: Materials are removed and mixed with other agents, or undergo a physical treament such as washing.

Natural attenuation: remove source of pollution and then monitor natural processes.

THEORY SECTION THREE

STATE OF THE ART - BIOREMEDIATION Bioremediation is an engineered technology that modifies environmental conditions (physical, chemical, biochemical, or microbiological) to encourage microorganisms to destroy or detoxify organic and inorganic contaminants in the environment.

The challenges for bioremediation technology have many dimensions, e.g.

1. speed, 2. tech/land area required Some of these challenges are entangled with social aspects, hence they are all not pure technological problems.

This workshop inspires a growing and active participation of designers, that are capable of reviewing these entangled constraints with the perspective of design; rather, designers when in collaboration with researchers, scientists, etc. might even turn these constraints into new opportunities, and parameters for creating a productive design mechanism.

POINTS TO REMEMBER 1. Metals cannot be destroyed through bioremediation technologies. Rather, microbes can remove dissolved metals from solution by reducing them to a more insoluble valence state.

2. Immobilization reduces the mobility of contaminants by altering the physical or chemical characteristics of the contaminant causing it to precipitate out of solution or to sorb onto the soil.

3. Furthermore microorganisms can mobilize inorganic compounds through autotrophic and heterotrophic leaching, chelation by microbial metabolites, methylation, and redox transformations

Photograph of a Heavy Metal Polluted Site in Varanasi, India Image Source: http:// www.livemint.com/ Politics/H6eL4JewrqqatlKmGQdnON/ How-microbes-canclean-polluted-waterfrom-drains-to-rivers.html

79

BIOREMEDIATION MECHANISMS

From Brownfields to Greenfields: A Field Guide to Phytoremediation

Image Source: http://urbanomnibus.net/2010/11/from-brownfields-to-greenfields-a-field-guide-to-phytoremediation/

DESIGN QUESTIONS FOR HEAVY METAL BIOREMEDIATION 1. Who will manage the Remediation technology ?

2. How will it be maintained or regenerated ?

3. What is the fate of the heavy metal ?

4. Can new value chains be derived from the recovery of pollutants ?

5. How do we prevent the heavy metal re-entering the food chain/ecosystem ?

THEORY SECTION THREE

DEFINING CRITERIA FOR BIOREMEDIATION TECHNOLOGIES IDENTIFYING FACTORS FOR TECHNOLOGICAL SELECTION

Reference: B.Parker: Biological Approaches to Land & Water Restoration. WCMT Report

As seen in Groundswell, Constructing the Contemporary Landscape, MOMA, 2005 Alumnae Valley, Wellesley College, Wellesley MA Designer: Michael Van Valkenburgh Associates Image Source: http://inhabitat.com/10-landscape-design-projects-that-turn-damaged-and-neglected-spaces-into-healthy-beautiful-environments/

81

Demonstration of Bioremediation Clean up Project using Microorganisms

Image Source: http://www.livemint.com/Politics/H6eL4JewrqqatlKmGQdnON/How-microbes-can-clean-pollutedwater-from-drains-to-rivers.html

THEORY SECTION THREE

BIOREMEDIATION WITH MICROBES BIOADSORPTION MECHANISMS FOR HEAVY METALS: CONVERSION TO A LESS SOLUBLE FORM Microbes are nature’s ultimate garbage disposal, devouring the dead, decomposing and inert material that litters Earth’s surface. They’re so good at it, in fact, that humans have taken an increasing interest in coercing them to clean up our environmental messes.

Many microbial detoxification processes involve the efflux or exclusion of metal ions from the cell, which in some cases can result in high local concentrations of metals at the cell surface, where they can react with biogenic ligands and precipitate. Although microorganisms cannot destroy metals, they can alter their chemical properties via a surprising array of mechanisms.

The main purpose of this session is to provide an update on the recent literature concerning the strategies available for the remediation of metal-contaminated water bodies using microorganisms and to critically discuss their main advantages and weaknesses.

The focus is on the heavy metals associated with environmental contamination, for instance, lead (Pb), cadmium (Cd), and chromium (Cr), which are potentially hazardous to ecosystems. The types of microorganisms that are used in bioremediation processes due to their natural capacity to biosorb toxic heavy metal ions are discussed in detail.

Microorganisms employed in the bioremediation and processes/mechanisms involved in the case of dead and living biomass.

Image Source: Luciene M. Coelho, Helen C. Rezende, Luciana M. Coelho, Priscila A.R. de Sousa, Danielle F.O. Melo and Nívia M.M. Coelho (2015). Bioremediation of Polluted Waters Using Microorganisms, Advances in Bioremediation of Wastewater and Polluted Soil, Prof. Naofumi Shiomi (Ed.), InTech, DOI: 10.5772/60770.

83

SECTION THREE

Workshop Participants during Day 2 of Design development

85

DESIGNING WITH NATURE SECTION FOUR

87

SECTION FOUR

ECO-CENTRISM NATURE CENTERED DESIGN DESIGN FOR AN EMPATHIC WORLD “Marx’s critique of ecocentrism reflects a practical-minded conviction that environmentalism must be anthropocentric (i.e. respectful of human interests) in order to be workable.”

- An Exchange on Thoreau Can we design around nature?

Our discussion on Nature-centered design tries to connect the scientific knowledge about the functioning of nature with user experience and human centered design approaches. We urge the participnats to imagine how design products, services, concepts for non-human living brings and see the impact of design and human activity on nature. The difference with the eco-friendly design done up to now, is that we will focus on the individual beings and grow empathy from them and not as natural process as a whole.

Image Source: http://farm6.static.flickr.com/5716/23209526236_f78f07709f_b.jpg

89

Image Source: http://farm8.static.flickr.com/7229/7353883248_0e80c81da3_b.jpg

SECTION FOUR

ECOCENTRISM as an ecopolitical route to save the planet Can we shift value systems from human focused to earth focused?

Does this counter what we are biologically predisposed to do?

Would we even want to do this?

Nature can live without humans, but humans cannot live without nature. Architecture can make this truth transparent and allow us to experience nature at a deep, transformative level. An important mission of green building and sustainable design is to bring architecture and urban planning back into the flow and cycles of nature. We need to reconnect buildings to their roots in climate, land, and place for current and future generations. We need to design with the understanding of our genetic need to be connected to living natural environments (biophilia). Designers and innovators need to not only reduce the obscene, mindless consumption and waste in the name of design, but design regenerative, living systems. We can make buildings and communities whole through commonsense design that incorporates life-enhancing technologies that incorporate the basic elements of sun, water, healthy landscapes, and clean air wherever possible.

Nature holds the key to our aesthetic, intellectual, cognitive and even spiritual satisfaction.

- E.O. Wilson

91

DESIGNING FOR NATURE Algae-Laden Hydrogels - Designing with Biomaterials BiotA Lab, 2016

93

THE DESIGNER’S APPROACH

References https://upload.wikimedia.org/wikipedia/en/thumb/0/0a/CK_Diagram.png/500px-CK_Diagram.png

THE BIG QUESTION: HOW MIGHT TECHNOLOGY BE DEPLOYED IN THE REAL WORLD? 1. Who will we manage the remediation technology 2. How will it be maintained or regenerated?

3. What is the fate of the heavy metal?

4. Can new value chains be derived from the recovery of pollutants?

5. How do we prevent the contaminant re-entering the food chain/ecosystem?

CONCEPT DEVELOPMENT

SECTION FOUR

CK (CONCEPT-KNOWLEDGE) THEORY The Concept-Knowledge Theory is a design theory based on the distinction between concept and knowledge, as its name suggests. Currently the theory is mostly used for explanatory purpose, but it can also be used as a very simple and powerful framework to generate innovative concepts.

NARRATIVE DEVELOPMENT -> PROTYPING AND FABRICATING SCIENCE While narratives allow us to explore science that can exist in different culture and context. We urge the participants to not just conceptualise but rather go one step ahead into visualising a prototype highlighting the same.

Why is it important to make something that can exist?

Why is it important for science to be designed?

We split in 5 groups where each group will ideate their project utilising the CK (Concept and Knowledge) theory. This session aims to connect the “ideas” from the participants to the “knowledge” accumulated during the workshop.

We decompose the parts of the concept like “what is cleansing/treating”, “who is the user”, “what is organism” to create alternatives to this concept and build the tree. This is an opportunity to see a wide range of properties/ideas for an initial ‘weird/odd’ concepts that you’re actually able to build something from your Kresearches. Guidance is provided during this session.

The core idea of this framework is to separate concept and knowledge in two different spaces, and to keep in mind that the object of study never has invariant definitions and properties.

A concept is defined as a proposition that is neither true nor false. It might emerge from market needs, that is when a technical or market requirement is not satisfied by existing solutions/technologies. Basically you can remember that concept = idea.

Concepts are gathered on the C-space.

Knowledge is defined as the group of propositions with known logical status (we know if they are true or false): all that we (or the designer) know belongs to this.

Knowledge is contained in the K-space.

95

Participants are inctroduced to the various materials / instruments / facilities available at the Institute of Making, to bring their ideas to reality

Special Training to use the machinery available at the Intitute of Making, UCL

DRIVING CREATIVITY WITH SCIENCE

SECTION FOUR

Now that every proposition can be placed, stored, or categorized in one space or the other, we will define 4 operations that can be used between these spaces.

Conjunction (C->K): When a concept/idea is tested in reality, we come to know whether the proposition is true or not (if the idea is feasible). Consequently, the propostion becomes part of the knowledge and leaves the concept space, resulting in an expansionof the K-space.

Disjunction (K->C): This is the operation by which a new idea/concept can be generated from existing knowledge.

Concept expansion (C->C): When an idea is conceived based on another idea, there is a concept expansion. It is important for concept expansion to support, incentive, visualize this process, and leave behind all consideration such as feasibility or other limitation. This expansion can be guided by other frameworks/ tools.

Knowledge expansion (K->K): This operation is the result of expansion of the knowledge by combination of it or new discoveries.

HOW CAN WE USE THIS? 1. Start expressing a issue/technical need and write it in C-space. You will probably base your concept on existing knowledge (solution/situation).

2. Express the existing knowledge associated. The solution here is not a lamp but the combination of technologies/knowledge.

3. With the knowledge, challenge the concept and formulate new ideas. You don’t have to challenge your concepts with knowledge, you can expand your concept using any method you want.

4. Proceed this way as long as you can: expand the two spaces by knowledge description and concept generation. Brainstorming and similar methods (see other frameworks!) might be used to enhance the efficiency of the concept expansion.

5. When you have finished (or when you are satisfied with the current ideas), test your concepts in reality, to try to operate a conjunction. During this stage, you will probably need to look whether some technologies or principles do exist, which will result in a knowledge expansion. In such a case: oh surprise! new propositions in K-space are available, and so you can challenge again your ideas/concepts. (go back to point 3).

6. If you have succeeded in operating a conjunction, you may have innovated!

Congratulations. Otherwise try to redefine your market/technical need in a more abstract way and restart the process.

A modelling of the original concept in terms of functionality may help.

97

DAY 3: INSTITUTE OF MAKING

99

Project 1: SELF IRRIGATE / LOWTECH BIOREACTOR

TEAM MEMBERS

SECTION FOUR

Florin Gheorghiu Giacomo Annio Pablo Izquierdo-Garrudo Lena Asai Shneel Malik

ABSTRACT Farmers in India have 2 - 3 crops a year. The problem is that the river is not very well maintained.

In Japan, rice fields irrigated with cadmium-contaminated water led to various diseases, such as skeletal deformation, hence a prototype for cleaning water used for irrigation by farmers in India was proposed.

On the river Yamuna, samples of water contained Cadmium over the accepted standards for both drinking and irrigation (Trace and Toxic Report of Indian Rivers, 2014) Chlorella vulgaris is shown to be effective in absorbing Cadmium from water in short periods of time.

Also, it is tested that dried biomass appears to be more efficient than live algae.

DETAILED DESCRIPTION A water treatment mechanism is introduced as an additive layer between the river output and the farmers crop land input, wherein, a ‘Bioencapsulated Algae’ remediation system is developed that treats the contaminated water before it enter the cropland.

This aims at primarily mass-adopting the mechanism of low-cost bioreactors that can be easily made available to a large percentage of farmer population.

SCIENTIFIC REFERENCES: Zeraatkar, Amin Keyvan et al. “Potential Use Of Algae For Heavy Metal Bioremediation, A Critical Review”. Journal of Environmental Management 181 (2016): 817-831. Web. http://www.cwc.nic.in/main/downloads/Trace%20&%20Toxic%20Report%2025%20June%202014.pdf Trace and Toxic Report of Indian Rivers, 2014

The Problem: High Level Toxic Metal Contamination in Yamuna River Concept: Bioremediation through Microalgae Knowledge: Bioencapsulation of Microalgae for Metal uptake Solution: Introducing a ‘cleansing system’ between the Farming irrigation system - wherein water from the iver is treated before being used to water the crops.

101

Project 2: BIO-BUCKET CHROMIUM BIO-REMEDIATION TEAM MEMBERS Ludovica Cantarelli Benjamin Denjean Francis Lister Maria Idicula Francesca Perona

A member of the team building a prototype of the concept at the Institute of Making, UCL

The Problem: Large -Scale Metal Pollution of water reservoirs by Tanning Industries Concept: To build filtration membranes within the system of metal disposal to stop water basins from getting polluted by metal contaminants.

Knowledge: Kombucha - an organic/degradable and recyclable filtration membrane.

Solution: Create a filtration membrane for toxic metals using Kombucha.

SECTION FOUR

ABSTRACT The solution focuses on using locally-sourced materials for a low cost modular filtration of chromium polluted water runoff from tanneries of India, following a “decentralize the mitigation, centralize the remediation” principle.

The set-up consists of stacked 200L containers containing a Kombucha scoby and each performing the filtration (Razmovski & Šćiban, 2008). Kombucha is produced using coconut water for pH and nutrients, complementary sugars are provided using molasses. The system is implemented at the outlet of each individual tannerie to limit the Cr flow to farmed drains, aquifers and downstream rivers. Cr is then reclaimed from the scoobies by submersion in a sulfuric acid bath.

To ensure uptake and correct usage of a novel technology by a variety of non-scientific stakeholders, the complex aspects of the technology (scoby manufacture, chromium isolation from the used scabies) are implemented by an external company.

This centralization of the system offers a number of advantages: 1. The company will ensure optimal control of growing conditions for the scobies, ensuring their correct function.

2. Potential secondary pollution issues in the chromium reclamation process can be mitigated in house.

3. Use of the technology by various stakeholders can be incentivized through a purchasing mechanism.

4. The repurchase of the scoby is conditional to the concentration of Chromium, thereby placing the company as a monitoring entity supporting local environmental agencies. The company operates out of the sale of Cr and treated Cellulose.

Further development need to address issues of - flow speed, - number of stages required, - possibility to filter other pollutant in the effluent and - robust economic analysis.

We argue that those can only be properly assessed through on-site pilot studies. Further improvement shall test different bacterial cellulose from directly available sludge (Cavka et al., 2013).

SCIENTIFIC REFERENCES: 1. Razmovski, R., & Šćiban, M. (2008). Biosorption of Cr(VI) and Cu(II) by waste tea fungal biomass.

Ecological Engineering, 34(2), 179–186. http://doi.org/10.1016/j.ecoleng.2008.07.020 2. Cavka, A., Guo, X., Tang, S.-J., Winestrand, S., Jönsson, L. J., & Hong, F. (2013). Production of bacterial cellulose and enzyme from waste fiber sludge. Biotechnology for Biofuels, 6(1), 25. http:// doi.org/10.1186/1754-6834-6-25

103

PROTOTYPING Creating a Filtration Membrane for Toxic metals using Kombucha

DETAILED DESCRIPTION

SECTION FOUR

The Ganga River is contaminated with heavy metals such as Cadmium (Cd), Lead (Pb) and Chromium (Cr); tannery factories pour their liquid waste in the river, filling it with pollutants.

Tanning consists basically of 2 processes: 1.

Leather transportation from one bath to another.

2.

Leather tanning During these processes, residual liquids are either released to the river or leaked in the soil.

Water from the river is often naively used by farmers as drinking water. Residues that leak in the soil filter to the aquifers and go directly to the municipality drinking water system.

Given this situation, the aim of our project was to create a filtration membrane for toxic metals using Kombucha.

Our area of interest would be a place that concentrates a lot on tanneries and our targeted pollutant would be Chromium.

Also, we want this solution to be cost-effective, so that farmers can afford the solution; for this reason we thought of using local materials that are available in the environment to build the system.

Kombucha would be grown with coconut water and blue bins would be used as filtration recipients. Water would be filtrated multiples and then the flow free of pollutants would come out from the bins. The medium for Kombucha could be grown by a side bioremediation company, that will monitor the experiment and clean the water, and the filtered Chromium could be after all re-sold to the tanner. This way both the bioremediation company and the tannery get a benefit from the action.

105

Project 3: CHROM-ACTION! REPLACING CHEMICAL ETPS WITH BIOLOGICAL ETPS TEAM MEMBERS Aleksandra Berd Linh Pham Karon Ng Yan Wah Adnan Arif Eamon Hassan Lina Salih

Explanatory Illustrations of Concepts emerging from scientific Papers - for sustainable implementation

The Problem: Unprecedented Increase in Chromium(Cr) Levels in River Ganges.

Concept: Bio filtration Process - altering the way industry sludge is filtered.

Knowledge: Kombucha - an organic/degradable and recyclable filtration membrane.

Solution: Biofiltration Products; urging active participation from the Community.

SECTION FOUR

ABSTRACT More than 170 tonnes of Cr is deposited through industrial activities and 40% of Cr used in tanning leather is retained in waste sludge. Vast amounts of industrial contaminants being poured untreated into the sacred waters. For example,Chromium levels are about 70 times the recommended maximum level (Banu Kamaludeen et al. 972-985). Cities and people use the river for laundry, cleaning, washing and religious rituals.

River Ganges is the biggest river in India and runs through 29 cities. It is considered the fifth most polluted river in the world and is considered sacred.

Tanning factories produce waste effluent through through industrial processes. Industry sludge currently treated by Effluent Treatment Plant with chemical and ineffective filtration processes. We propose a new biofiltration process.

We also propose a new way of connecting citizens who suffer from Cr pollution and the rest of the world. The necklace allows citizens from all over the world to create a personal relationship with communities who live in areas suffering from Cr pollution. An object can be a constant reminder of what is happening in India.

DETAILED DESCRIPTION One of the biggest problems of the area is the Chromium (Cr) contamination. Multiple industries, mainly leather tanning ones, release their wastes to the river, thus polluting the water and provoking several health effects. Considering the cultural tradition of the Ganges River, it is of crucial importance that the water is free of pollutants, toxic Cr forms included. For this, our idea is to slightly change the way industry sludge is filtered in the Effluent Treatment Plants (ETPs). Currently, they use multiple chemical compounds to help cleaning the water, but these, together with the filtration systems, are not effective enough.

Thus, we propose an implementation of a biological filtration system in the ETPs; a biological membrane containing microorganisms able to capture the Chromium from the water. Once filtered, water will be Cr-free and the captured Cr could be used by the NGO to build products such as jewelry, which on one hand makes the project sustainable and economically valuable and on the other hand it is an inspiring idea for people to make products that could serve as merchandising in order to crowdfund the project.

SCIENTIFIC REFERENCES: 1. Banu Kamaludeen, Sara Parwin et al. “Bioremediation Of Chromium Contaminated Environments”.

Indian Journal of Experimental Biology 41.September, 2003 (2017): 972-985. Web. 13 Feb. 2017.

2. “Tannery Effluent Treatment”. leatherpanel.org. N.p., 2017. Web. 13 Feb. 2017.

107

Project 4: CITIZEN-LEAD (Pb) DETECTION TEAM MEMBERS Ariana Mirzarafie-Ahi Waldemar Matuska Mourdjen Bari Simona Della Vale

Members of ‘Citizen-Lead’ Detection project presenting their prototype at the Workshop

The Problem: Lead contamination of Soil can lead to serious health problems; through ingestion and inhalation.

Concept: Lab-on-a-Chip: Crowd - sourcing; involving locals to collect Lead samples and detect percentage of contamination Knowledge: Metal Uptake Experiments - Learning to Code and build devices that can be easily distributed for use by locales - urging a growin activity of both learning and active feedback within the established system Solution: Citizen Led - Data Collection Device for detection of Heavy metals

SECTION FOUR

ABSTRACT Lead contamination of soil can cause lead poisoning which can cause a wide range of symptoms. Poisoning occurs through ingestion and inhalation. Detection of lead in the environment could be a key step in preventing health issues.

Acid-sodium rhodizonate lead detection products such as D-lead by ESCA Tech Inc. exist for quick lead detection and the presence of lead causes a visible purple colour.

Locals can use wipes on surfaces inside their home as well as outside in the field in order to check for lead presence. They get an instant result as well as feedback on actions they can take such as telling children to avoid contaminated area. Their data can be collected remotely via a smartphone app or separate standalone device. The resulting data of positive lead tests and associated location and time can be used by NGOs, govt. and others to raise awareness in real time and begin setting into motion decontamination efforts. This is achieved through data recording via smartphone app or standalone device.

DETAILED DESCRIPTION Dust of lead represents a real danger for population. It exists almost everywhere, from kitchens to playgrounds, but most of the times people is not even aware of its existence, and there exists no data aiming to map polluted areas. Considering that most of the population owes an android smartphone, the idea of the present project would be to create an android application that could both detect lead contamination and send this data to a server in order to geolocalize it. This way you make people know where the contamination but also you make the government realize about the severity of the problem.

The technical description of the project is the following: locals would be provided with a set of lead detection papers. These papers, in contact with the lead, turn into purple color. They would select a specific place and scratch the ground with the paper; then they would open the mobile App and select the option take a picture of the paper, which may have changed its color after the contact with the ground; the App would ask them to select a point in the picture and then will calculate the color in the RGB scale and send the data to a server, that would collect both the coordinates (which would be placed in a map of the area) and the RGB number. This is a really interesting citizen science project, as you are empowering locals to detect, measure and geolocalize contamination themselves; later on you could give them a kit to bioremediate their own soil in case it would appear to be contaminated.

SCIENTIFIC REFERENCES: 1. Zhao, Liyun et al. “Fluorimetric Lead Detection In A Microfluidic Device”. Lab on a Chip 9.19 (2009): 2818. Web. 2. Satarpai, Thiphol, Juwadee Shiowatana, and Atitaya Siripinyanond. “Paper-Based Analytical Device For Sampling, On-Site Preconcentration And Detection Of Ppb Lead In Water”. Talanta 154 (2016): 504-510. Web.

109

Project 5: FUNGI EDU KIT TEAM MEMBERS Alexandra Carr Fabienne felder Louise Rymell Madeleine Stack

Members of ‘Citizen-Lead’ Detection project presenting their prototype at the Workshop

The Problem: Large-Scale Lead (Pb) Contamination of Soil and water Reservoirs.

Concept: DIY Science Kit Knowledge: Fungi Paecilomyces javanicus and Metarhizium anisopliae have been found to transform lead metal into the mineral ‘chloropyromorphite’ - the most stable and non-toxic form of lead.

Solution: Reducing Lead toxicity in contaminated Reservoirs through active participation of Locales via a ‘Fungi DIY Kit’.

SCIENTIFIC REFERENCES: 1. RHEE, Y J, et al. Lead Transformation to Pyromorphite by Fungi, Current Biology 22, 237–241, February 7, 2012 2. SAYER, A J, et al. Lead mineral transformation by fungi, Current Biology, Current Biology 1999, 9:691–694

ABSTRACT:

SECTION FOUR

The fungi Paecilomyces javanicus and Metarhizium anisopliae have been found to transform lead metal into the mineral ‘chloropyromorphite’. Chloropyromorphite is the most stable and non-toxic form of lead “and the various forms of pyromorphite (Pb5[PO4]3X [X = F, Cl and OH]) formation have been widely proposed as a remediation mechanism for the sequestration and immobilization of contaminant Pb in soil” (Y. J. Rhee, et al., 2012) We see a potential in using these kinds of fungi in bioremediation, either by cleaning contaminated soils, but potentially also to decontaminate elements in industrial processes, respectively use them to extract lead from a structure, before they reach soils and water (e.g. to “infect” objects like batteries, etc. to neutralise and recycle them. Other fungi (e.g. A. niger) have been found to turn insoluble pyromorphite bioavailable, i.e. soluble”).

In this project we propose an educational tool that could help communities, farmers, etc. decontaminate pieces of land using M. anisopliae. M. anisopliae occurs in soils all over the world. We provide them with the appropriate fungus or teach them to identify it at home, and provide them with instructions on how to use it and education around it.

The soil needs to incubate for a least several months for the process to take effect. There are some worries using this fungus as it is harmful to some insects. It has, however, also been used as an insecticide and is even under study to have positive effects on the control of malaria transmitting mosquito populations.

As with every fungus, it tends to spread and we have not found a solution how to contain it or control it/kill it off. Some ideas were considered, such as burning the fungus, rendering it sterile or introducing other lifeforms that control the fungus after it has performed its job.

It might also be, that the fungus can just be left alone, as it’s found in many times of soil around the world already.

DETAILED DESCRIPTION There exist 2 types of fungus that are able to transform Lead (Pb) into a non-toxic more stable form of this heavy metal; this being a possible potential solution to bioremediate Pb polluted environments. Within this context, we can talk about different application possibilities: either we could remediate already contaminated soil or decontaminate objects or elements before Pb contained in them reaches the water.

Being inspired by the concept of Citizen Science, we have thought of empowering locals to incubate these fungi into a soil sample and later on spread them out in their fields. The format we imagine our project to have would be a kit-box containing all the needed materials to grow the fungi (seeds, macro and micronutrients, etc.) as well as detailed and clear instructions to do so.

Still, there is still some troubleshooting to be done, as this fungi can be beneficial for soil in terms of cleaning contamination, but at the same time toxic for its living ecosystem, this is, for some insects living there, fact that could alter the food chain within the ecosystem. Possible solutions could be to cover the area before decontaminating, to burn it or to use genetically modified fungi pre-programmed to kill themselves after a concrete time lapse.

111

CONCLUSION CONCLUSION SECTION FIVE

113

SECTION FIVE

Learning outcomes and good practice from organizers Open Science School’s Co-lab Bioremediation

1. EXPERIMENT: The protocols were very well designed. We can see a clear link between the experiments conducted at the workshop and the theme of the workshop.

This is the first time in Co-lab where we worked so closely with a laboratory. Having Brenda and Laura on the project execution team allowed the organisers to create a programme which had such a strong connection between the theme of the workshop and the content of the experiments. The practicals in the lab gave participants to get a deeper understanding of the problem that exists in India.

The workshop that takes time to show results, like Kombucha, might want to be avoided in the workshop. We may want do experiments that are faster. We can see the phenomena happening at the workshop. For things that take time, we can show a video.

Our original idea was to show that experiments can be done in the lab - it can be done anywhere by anyone. We need to question what role science plays in a workshop. There are so much to learn beyond the workshop. We have been using science as source of information - This kind of approach is reflected in our lectures - we show what is science rather than demonstrating science.

For future workshops, allowing participants to engage in thought experiments or discuss the predicted outcomes of experiments before entering the laboratory environment may encourage playfulness and inclusiveness among scientists and designers.

2. DESIGN / IDEA GENERATION: We felt that the entire concept behind CK Theory was not exploited to the best of its abilities.

The flow of the workshop was good up until the end of the workshop. Design became a very mechanical process. We realised that ‘a protocol for design’ does not work in a workshop context. We should provide an open ended question.

The organisers focused their effort in making the science an opportunity to understand the problems present in India. The design part did not investigate a deeper connection with the questions. Perhaps, we should introduce ​elements of design of rather than theorizing design.

We, as facilitators, need to think in more depth on how we share design, as there is no strict protocol on how to do design (there will never be).

Design thinking was present in the planning of the workshop and this is why the workshop flow was good. However, the adaptation of the workshop adaptation of knowledge gained by the participants was not translated in a way expected by the organisers of the workshop.

One of the reason was we expected too much from the 3 days. It is demanding to ask someone to create an idea. In the future we need to focus on the idea generation rather than asking the participants to create an idea from scratch in 3 days. This process often makes the ideas generic and predictable.Creation of new ideas, may not be the most essential elements/expectation organisers of the workshop expect from the participants. We should focus more on disseminating the true philosophy of Co-lab.

115

Learning outcomes and good practice from organizers Open Science School’s Co-lab Bioremediation

3. PROTOTYPE: Thanks to the facilities provided by the Institute of Making, the participants were able to work on the prototyping, however, we must admit that is it demanding to ask the participants to learn a whole topic from day 1, generate an idea for a project by end of the day 2 and finish a prototype and present the work by day 3. The work that comes out of the workshop is very sophisticated, and in reality projects from Co-lab has developed and received research grants or exhibition opportunities. What is evident is that the strongest project did not necessarily succeed becomes the prototype was very successful. It is because the ideas were strong. In order for the participants to truly gain from the use of the workshop facilities, the organisers must plan a programme to generate stronger ideas by the end of the idea generation phase of the workshop.

We are not dismissing the concept of thinking by making - it is a very important methodology practiced by designers. Instead of just simply bringing all of the making at the end, we should bring back more making in all of the phases of the workshop. Understand by making, think by making and generate ideas by making. This workshop focused a lot on ‘understanding by making’, which worked fantastically in the laboratory. In future, we should introduce more activities to help participates generate ideas by making, rather than expect them to have an idea before they start making.

Possible Suggestions for Future Co-Labs’

SECTION FIVE

Open Science School’s Co-lab Bioremediation 1. ADDING (OPTIONAL) DAYS 4 & 5, TO DEVELOP PROTOYPES: If we continue the Concept generation / Idea formulation part of the workshop from day 2 into the first half of day 3, then the participants might get more time to generate and formulate their ideas. Further, before the end of day 3 - the participants are inducted into the Institute of Making allowing them to explore the facilities provided by UCL into making and actually visualising their prototype.

We could potentially offer a day 4 & 5 (optional) wherein, the facilities are duly made available to the teams by the institute of making in order to guide and provide them support into ‘making’ their prototypes. It becomes important to allow the participants enough time into absorbing the information load provided to them during the first half of the workshop - building on constructive design and visualisation thought processes - putting into existence the knowledge gained. The notion of ‘rushing’ through the process - pushing the participants to generate a prototype at the end of day 3 might be unfair - whereas, this would allow the workshop to ‘willfully’ continue into day 4 & 5, provided the enthusiasm of the participants remain intact.

The idea is to maintain the interest of the participants to not just the 3 constrained days of the workshop, rather to intrigue them into continuing the learning process that has just commenced through their active participation / involvement into the Co-Lab. This reflects a lesson the Co-lab team learned from the Co-lab OpenPlant series (Workshop #7), where split the workshop into a 3 independent sessions and picked the most motivated 3 projects to fund to prototype in another 3 days session. The ideas that came out of the ideas were definitely more concrete as participants spent 3 days to create the idea. The problem, however, was that the sessions were spread out across 3 months, which deteriorated the motivation of the participants. Through this series we realised that participants definitely need more time to come up with an idea, but we have to start making immediately after the idea generation phase of the workshop.

2. EXPLORING DIFFERENT WAYS OF KNOWLEDGE DISSEMINATION (FROM THE EXPERTS): Collecting ideas and thoughts of people / experts from the Industry who have or are trying to execute a similar scientific research or working in the field to tackle the same problems - asking them about their experience over the years of their practice, learning from their field related observations, the processes that they undertake and most importantly what they would expect from an ‘interdisciplinary’ workshop in the true sense. Then connect their knowledge through interviews from designers who are increasingly collaborating with clients and researchers from the field - sharing their experiences, and design related difficulties and some thought provoking suggestion for the participants of the workshop in guiding them through the next few days of the intensive collaborative week of knowledge - gaining and idea development. However, it is worth reflecting as to when is the most appropriate stage to bring in expert inputs, for maximum input ‘balancing the integration of knowledge without overwhelming the participants’.

This proposition allows the organisers to dig deeper into the topic of the workshop - starting to getting to know the topic and the people involved in the research.

117

All Participants brain-storming to develop prototypes around there learnings from Days 1& 2 of the Workshop

119

CREDITS ORGANISERS OPEN SCIENCE SCHOOL/DITOS AT CRI Lena Asai Imane Baïz Juanma Manuel Garcia Arcos Ke Fang Alicia Mansilla Sanchez Elina Moraitopoulou Mourdjen Bari Anirudh Krishnakumar UCL Brenda Parker Laura Stoffels Paloma Portela Shneel Malik iGEM TEAM 2014 Xenia Spencer-Milnes Despoina Paschou Alex Dack Haroon Chughtai PURE EARTH, BLACKSMITH INSTITUTE Promila Malik David Hanrahan CENTRE FOR ENVIRONMENT EDUCATION Reema Banerjee INSTITUTE OF MAKING, UCL Olivia Clemence Ellie Doney

CREDITS PARTICIPANTS Florin Gheorghiu Giacomo Annio Pablo Izquierdo-Garrudo Ludovica Cantarelli Benjamin Denjean Francis Lister Maria Idicula Francesca Perona Aleksandra Berd Linh Pham Karon Ng Yan Wah Adnan Arif Eamon Hassan Lina Salih Ariana Mirzarafie-Ahi Waldemar Matuska Mourdjen Bari Simona Della Vale Alexandra Carr Fabienne felder Louise Rymell Madeleine Stack

121

CO-LAB

BIOREMEDIATION 26-28 Nov 2016