By Braden Milford, U.S. Stockholm Junior Water Prize Winner

My name is Braden Milford, I am 17 years old, and I am from Tulsa, Oklahoma. I am often caught up in schoolwork, but when I am not studying, I love to spend my time outdoors camping, backpacking, skiing, biking, climbing, scuba diving, and just about anything else. I am an Eagle Scout and really feel happy in the outdoors, immersed in nature.

This is where my passion for the environment comes into play. I have been fortunate enough to travel around the country and world and see many different places and cultures, as well as the wide range of ecosystems and environments our planet has to offer. One part of loving nature and the outdoors is getting out and enjoying it, but a second and perhaps more important point of loving the outdoors is protecting it. It is clear that for every breathtaking view, there is just as much destruction elsewhere in the world. There is plastic debris in our oceans, acid rain in cities, and heavy metal contamination in drinking water.

I have completed three years of high school, and each year done a different project. Yet, they all had something to do with water. During my freshman year when I decided I wanted to do research, my science teacher offered that I could research anything I wanted to. There were kids working with cancer and drug discovery, computer programming and machine learning, animal behavior, and gene editing. All of these areas seemed interesting enough, but none really excited me. My teacher then proposed the area of environmental sciences and started talking about water toxicity. I was really interested, given my passion of the outdoors as well as all of the prominent water problems our planet faces, none of which seem to have a good solution despite the numerous years these problems have been evident.

My freshman year project was originally to study how ocean acidification would affect ocean species, and I was going to do this by watching sea urchin development in different pH concentrations of water. I was introduced to the true world of research right off the bat, however, after a month of preparation, all of my sea urchins turned out to be either male or infertile due to a cold blast during the shipping process. After two shipments of sea urchins and two weeks left until the regional science fair, I moved onto a new project idea, this time broadening my area of interest to aquatic acidity in general. My new project was nothing groundbreaking by any means; I studied how acidic water effected worm regeneration, plant growth, cell membrane integrity, and enzyme function, but this project was really the first step in the journey that led to me winning the Stockholm Junior Water Prize. I would say the failure of the sea urchin project was one of the greatest things to happen, because I was forced to expand upon my interest in order to have a project that I could present at the regional science fair. This freshman year project was important because it allowed me to find my passion. I realized I could really care less about working with cancer or learning to code; there were so many problems in our water supply, and nobody can escape them. Water is a unifying force of sorts-absolutely all life demands it. If water is not freely available and in a clean state, all life will suffer.

My sophomore year project saw my passion for water research grow. I worked with the fire retardants dropped out of planes to contain wildfires. I decided to move away from environmental acidity because it seemed like a crowded field, but fire retardants had never really been studied at all with the exception of the basic EPA studies. This was a brand new topic that I could investigate for the first time. There was no sense of how the retardants would affect model organisms or stream water quality. In my freshman year project, it was very easy to hypothesize what acid would do to seed germination, but I think I was even more interested in the fire retardants because of the fact that there was little to no background information to predict what would happen. This project also allowed me to get my first exposure to field research. I was able to go and study a stream in Colorado that was in the middle of an active wildfire, where fire retardants had been dropped. This experience perfectly partnered my love for nature and exploring with my love and interest for my research. I focused on the water quality (dissolved oxygen, pH, temperature, conductivity, turbidity, nitrates, phosphates) of the stream and how the fire retardants affected it.

As I mentioned, this sophomore project allowed me to grow in my passion for water research, but I was also able to grow as a researcher, a problem-solver, and a thinker. After extensive toxicity testing with the retardants, I decided that anyone can point out a problem and study it, but the real value in this day in age of research is finding solutions. We always hear about the problem, but rarely the solutions, and that is because it is much more complicated to fix a problem rather than say it exists. So, I spent three months working with cyanobacteria and the process of nitrification to develop a potential solution to remediate the fire retardants out of contaminated streams. This project was the second major step in getting to the successful place I am now. I had a solid interest after my freshman year, but this sophomore year project allowed me to grow, discover, and think in new ways, as well as get exposure to what my peers were doing with water research.

In early June of 2017, I was ready to get right back to work. I spent a lot of time debating whether or not to continue my work with fire retardants, specifically the remediation system I had developed. I had designed and tested a system that worked to some extent, but I felt very limited in what I would be able to do beyond that initial design. I didn’t quite understand how I would go about refining the design and implementing the system, so I ultimately decided to move on to a new area. I did keep my key interests from my previous projects at the center of what I wanted to do, however, so I knew I wanted to work in the field and also focus on fixing problems. I also wanted to choose a problem that was widespread around the world, because if I was able to succeed in creating some system that can help attack a problem, I would want people everywhere to have easy access to the clean water I could hopefully create.

I decided to study heavy metals that are a result of acid mine drainage found at the millions of mines across the world. In the U.S. alone, there are an estimated 500,000 abandoned mine sites, and only about 1,332 of these are part of the EPA Superfund Cleanup program. This is mainly because of funding reasons, as the EPA spends about $300 Million annually on the Superfund program, meaning each mine site takes hundreds of thousands of dollars to maintain.

I read hundreds of sources and began brainstorming ideas for my project during the summer of 2017 and eventually came to my idea: I wanted to create a cost-effective, low impact system that can remediate heavy metals out of contaminated water systems. The cost-effective point was important because by lowering cost and at least maintaining efficiency, if not improving it, the $300 Million used annually could be used to increase the number of contaminated mine sites in the Superfund program. The low impact point was key because I didn’t want to leave any permanent machinery or building at these mine sites; I wanted to create an easy to remove system that will not have a lasting effect on the environment long term. 

One final conclusion I came to was that I wanted my system to use bioremediation rather than building technology to do the work. I figured that the most environmentally friendly solution would be to use the organisms already living in the environment I wanted to fix.

I was able to start physical work on my project in mid-September 2017 at a field study at the Pennsylvania Mine Superfund Site near Dillon, Colorado. I worked with water samples from this site and another field study done at Tar Creek Superfund Site in Miami, Oklahoma through the fall, specifically focusing on the bacteria I collected in water samples from different points along the contaminated streams. 

I ended up having 250 environmental strains of bacteria that I isolated from the water samples, and I spent much of December and January screening the bacteria for successful growth in differing heavy metal concentrations as well as successful biofilm formation in differing heavy metal concentrations.

As I mentioned, I wanted to use bioremediation to remove the heavy metals from the water systems, and after extensive research, I found that when bacteria form biofilms, they possess certain mechanisms that allow them to intake heavy metal ions. So, after my screening work, I selected 24 of the 250 bacterial isolates that showed the greatest potential to successfully remediate heavy metals.

To design my system, I didn’t want to simply throw all of the bacteria together into a common culture. Each genius and species of bacteria obviously has different DNA and could therefore be suited to remediate some metals better than others, so I decided it would be necessary to identify my 24 bacteria so that I could best group them together when designing my system. To do this I was able to run a 16S Ribosomal Subunit Analysis of my bacteria, which focuses on isolating and sequencing the 16S gene in each of my bacteria’s DNA. I could then take the gene sequences from my bacteria and plug them into the online BLAST database, which matches my bacteria’s gene sequences with already identified bacterial gene sequences so that I could identify the bacteria.

Once I had identified my bacteria, I could group them by genera (in my sample I identified 6 genera and 11 species). For my remediation system to work, I needed the bacteria to form biofilms. For biofilms to form, the bacteria would need a food source. So, I decided that partnering my genera of bacteria with mixed green algae would allow the bacteria to use the algae as fuel to begin biofilm formation. Secondly, it was crucial that non-native species were not introduced into streams, so I needed a way to contain and immobilize my algae and bacteria cells. To do this, I found a substance called Sodium Alginate. Sodium Alginate is clear and jelly like, and I could easily distribute algae and bacteria cells within it. I could then suck up the mixture in a standard dropper pipette and drop the mixture into Calcium Chloride, which acts like a seal and forms a sort of “bead”. The best way to describe what these beads look like would be to compare them to the Orbeez kids toys or to the Boba that you put on frozen yogurt. They are small spheres, but algae and bacteria cells are fully contained within them and the bacteria can form biofilms and intake heavy metal ions from the streams. 

In my initial testing, I produced 600 of these beads, and the cost was less than $10. Of course, scaling the system to fit a real stream will see costs increase, but the cost for this system is still so much lower than current methods that this system would still be groundbreaking. Secondly, within just an initial two-week testing period, the majority of my systems were able to remediate over 80% of 5ppm concentrations of seven different heavy metals, a rate unseen in terms of heavy metal remediation. 

Obviously, my system turned out to be a success, and I see the source of this success to come from a lot of places. First, my skills as a researcher were crucial. My first two projects were instrumental in helping me find what I was interested in as well as learn how to ask questions and go about solving problems. Secondly, I put in a lot of hard work. From the initial brainstorming session I had in June of 2017 to where I am now, I am going on 13 months of work, as I am still running tests. I have gone to school at 7:30am in the morning, completed my day at 3:00pm in the afternoon, worked in my school lab until 3:00am in the morning, gone home to do homework, and be back at school at 7:30am the next morning. Finally, I would say a little bit of luck. I was fairly certain I would get some sort of success with my system, even if it wasn’t extremely efficient, but I never could have predicted the majority of my systems taking in over 80% of the heavy metal concentration in just two weeks, and I don’t think any professional scientist would have predicted this either. I think to have a sort of “scientific breakthrough,” you almost just have to be lucky. These things can’t be expected by any means.

And that brings me to the Stockholm Junior Water Prize. I actually got the invitation to apply to the State competition from my regional fair for my sophomore year project with the fire retardants, but I didn’t win. I got the invitation again for this junior year project, and I applied to the State competition and this time I won. I actually think I was more nervous for the national Stockholm Junior Water Prize Competition than other national and international competitions because SJWP focuses only on water, and I would be going up against the absolute best in specifically water research from around the country, not the normal broad category of environmental sciences.

What I quickly found at the competition, however, was that the national SJWP was not simply just a competition. It was an opportunity to not only make friends, but to collaborate and get support from peers and professionals alike. We all had extremely great projects and ideas, and I think what I took away from the event the most is not the competition, but the desire of everyone to make a positive impact on the water world. The students, judges, and coaches want to see all of these ideas succeed to the point that they are not being presented on a tri-fold board, but implemented in the field making a difference. I have made friends at several national and international science fairs, but I feel like I especially connected with my friends from the national SJWP. Everybody researches water, so everybody has something in common. I also think the general spirit of everybody at the event helped make it so special. Everybody wants to help everybody succeed. Evelyn from Hawaii and Avni from Minnesota want to help proof my poster before the international competition. Anjali from Kentucky offered to help get me into 3D Printing as I brainstorm for the future of my project. We actually have a group chat of about 8 of us that talk every day. We all also seem to have very similar tastes in music and share recommendations every day. 

In the days following my victory at the national SJWP, I felt like the floodgates of support and guidance opened. Professors and professionals from all around the country contacted me offering guidance and support, and I think that is the biggest thing I am taking away from this experience. My one worry after finding the success of my system has been how to go about implementing it. I didn’t want to leave it behind like I did with my sophomore fire retardant project. I think the most valuable thing about SJWP is by far the support, advice, and guidance I have and will receive as I now continue my project and develop it so that it can one day be implemented.

Braden's Freshman Year Project being displayed at the Regional Science Fair.

 

 

Braden's Sophomore Year Project that allowed him to grow his passion for water research.

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