The Effect of EV Battery Metals on River Microbial Systems

Table: ENV4
Experimentation location: Reseach Institution
Regulated Research (Form 1c): No
Project continuation (Form 7): No

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Abstract:

Bibliography/Citations:

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Additional Project Information

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Research Plan:

The research will investigate the impacts of common EV Battery Metals on River Microbial Systems. 

First, a water sample will be taken from Carnegie Lake near Princeton, NJ from a point roughly 20 feet from the shore, off a pier that leads into the lake. Collection will occur by dipping an empty collection bottle and capping. This water will then taken to the lab and stored in a cold room that maintains a temperature of -4C. In order to mimic the conditions of the river microbial system, the water will not treated in any way. Prior to using the water, the bottle will be shaken to ensure a uniform distribution of bacteria throughout the sample. 

The metals that will be used in the study are Lithium, Nickel, Manganese, and Cobalt in the forms of LiCl, NiSO4, MnCl2, and CoCl2 respectively. Controls of NaCl and NH4SO4 will be used to ensure that the secondary ion in the compound was not responsible for any effects used. Metal ions and Nutrient agar will be mixed at varying levels of concentrations of 0.1M, 0.01M, and 0.001M, with 3 plates of each concentration and chemical. Positive and Negative control plates were also created. Later, the water samples were taken and they were plated. While plating, a lab coat and gloves will be worn to ensure that none of the agar or chemicals have contact with skin. Following this, they will be stored in a room temperature storage at a lab in the University of Pennsylvania Biomedical Research Building and left to grow.

After a time period of a few days (3), the plates will be examined, and a count of the number of colonies will be conducted. This count will be compared in order to measure the toxicity of the metals tested. 

The plates will then be disposed of in a trash for Biohazards, this trash is then autoclaved and sterilized before being properly disposed of.

 

 

 

Questions and Answers

1. What was the major objective of your project and what was your plan to achieve it? 

       a. Was that goal the result of any specific situation, experience, or problem you encountered?  

In the aftermath of the 2011 Japanese earthquake, carcinogenic fallout from the Fukushima nuclear-plant contaminated groundwater. Having lived through this incident, the value of clean water became apparent and understanding the science behind toxicology and environmental pollution, with a specific focus on its impact on human-health, consumed me. 

In my sophomore year, it seemed that no matter where I turned a Tesla or other EV would be looming. My curiosity piqued, I discovered upon further research, that the rise of EV sales have been exponential in the last few years. But I also uncovered that, while EVs have been touted as safe, environmentally friendly alternatives to traditional cars, the reality is not as ideal as it may first seem. I was obsessed over the question “What are the impacts of EVs on the environment?” I discovered that Over 90% of EV Batteries are not recycled, and many are instead often discarded in hazardous, environmentally unfriendly conditions. This extends to Lithium Ion batteries as a whole, as 98% of Lithium Ion Batteries end up in landfills. 
 

       b. Were you trying to solve a problem, answer a question, or test a hypothesis?

 Through my research, I hoped to answer the question “What are the impacts of EVs on the environment?”I focused my attention onto the impacts of these batteries on local river systems. Specifically, the effect of the battery ions (Li, Mn, Co, etc.) on river microbial systems, which play a vital role in river ecosystems. The results indicated that almost all the EV metal-ions, especially Nickel, had an adverse impact on bacteria growth. 
 

2. What were the major tasks you had to perform in order to complete your project?

       a. For teams, describe what each member worked on.

This was an individual project under the mentorship of  Dr. Jefferey Field at the University-of-Pennsylvania (Department of Toxicology). My main duties included designing the experiment, collecting samples, creating solutions containing the ions needed,  producing agar plates mixed with the ions, plating the bacteria and collecting the data. 

Specifically,  I investigated the impacts that common metal ions, such as Lithium and Nickel, found in EVs and other batteries had on river microbial systems. As part of the research, I cultivated bacteria taken from a river in both a control sample as well as a sample containing metal ions commonly found in EV batteries. After incubating these samples, I then sequenced the DNA of the bacteria and analyzed them in order to study the effects of the EV ions on bacterial growth. Though further research is required, the results indicated that almost all the EV metal ions tested had an adverse impact on bacteria growth. In particular, the data indicated that Nickel had the most debilitating effect on the bacteria, almost completely preventing their growth. These experiences strengthened my foundation in research as I was able to learn vital skills while working in the lab, such as lab safety, growing Agar (bacteria) plates, plating and incubating bacteria, isolating the 16s rDNA, and finally running the 16s rDNA through an Oxford Nanopore Flongle for sequencing and microbiome analysis. 
 

3. What is new or novel about your project?

       a. Is there some aspect of your project's objective, or how you achieved it that you haven't done before?

Through this project, I was introduced to methods of growing and plating bacteria, as well as the Oxford Nanopore technology to sequence DNA allowing for greater analysis of microbial systems. 

       b. Is your project's objective, or the way you implemented it, different from anything you have seen?

While a few papers have analyzed the composition of Lithium batteries in the context of potentially harmful materials, none have linked them to the impact of the rising EV industry on river microbial systems. 

       c. If you believe your work to be unique in some way, what research have you done to confirm that it is?

In preparation for the project, I scoured the internet looking for research papers that could aid me in designing and researching the experiment. While some papers have analyzed the composition of Lithium Batteries in the context of potentially harmful materials, little to none have touched upon the impact of EV Batteries on microbial systems. From my relentless searches, I found one paper written by two college students that analyzed the impact of Lithium on the respiration rates of microbial rates, however they did not analyze other, more toxic, metals from Lithium batteries such as Cobalt or Nickel, and furthermore they did not analyze growth rates of these microbial systems. Thus, I believe that my research and subsequent analysis are unique and have potential implications for EV Batteries, as analyzing a wider variety of metals gives a better picture as to the impacts of EV Batteries as a whole rather than one specific metal. 
 

4. What was the most challenging part of completing your project?

      a. What problems did you encounter, and how did you overcome them?

Every worthwhile pursuit encounters obstacles, and my experiment was no exception. The path to achieving our final results was marked by twists and turns. The biggest problem I encountered in my experiment was that the agar plates liquified in my early trials of the experiment. This means that they did not solidify correctly and thus I was unable to use those plates. Additionally, the agar plates were difficult to work with, as after the agar solution was autoclaved, there was only a limited time window to create the agar-ion solutions and pour them into the 50+ plates. Initially, I suspected that one of my ions had some sort of reaction with the agar preventing it from solidifying, as a majority of my plates that liquified belonged to the Mn-ion group. However, after further research and reaching out to my mentor as required, I realized that the issue was that the distribution of agar (which was the component that made the solution solidify) in the nutrient broth was uneven, causing some plates to receive more agar than others. In order to overcome my issues, I used a systematic approach, ensuring  to mix the agar solution very thoroughly before pouring them into plates, allowing all the plates to solidify. I also developed an efficient system for pouring multiple plates quickly.

      b. What did you learn from overcoming these problems?

 In overcoming these problems, I learned to keep persisting in problem-solving, as I persisted in finding solutions despite initial setbacks in order to achieve success. Although I initially was dejected after thinking that I couldn’t use one of the ions in the experiment, I kept going to try and find a way to make the project work, reaching out to peers and the professor as required. This persistence not only allowed me to  identify the true cause for my difficulty but also enhanced the overall accuracy of my research by enabling the testing of  a variety of ions and in preventing this issue from recurring in the future.


While I managed to cultivate the bacteria successfully this time, I encountered a new hurdle when I couldn't sequence the DNA. Undeterred, I took it upon myself to learn some basic  Nanopore sequencing techniques independently. When I hit roadblocks, I sought guidance from peers,  my assigned mentor and the professor. On professor’s suggestion to account for any inaccuracies in nanopore sequencing results I used techniques to count the bacteria colonies on the agar plates.

Overall, overcoming these obstacles not only strengthened my research but also reinforced the importance of resilience and perseverance in achieving success.
 

5. If you were going to do this project again, are there any things you would you do differently the next time?

If given the opportunity to revisit this project, I would approach it differently based on the knowledge and experience I have gained since its inception. When I first began exploring the impacts of EV batteries on microbial systems, I was relatively new to battery research. To investigate this, I utilized solutions containing EV battery ions and observed their effects on agar plates. 


In future iterations of this project, I plan to employ a more direct method by testing Lithium Ion batteries directly on river microbial systems. While this experiment poses inherent risks due to the reactive nature of lithium with water, including the potential for explosions, the insights gleaned from this research would be invaluable in understanding the environmental impact of EV battery disposal.


Reflecting on my journey thus far, I believe that despite my initial approach, the experiment design I formulated was sufficiently applicable, as evidenced by the questions it raised regarding the EV battery disposal process and its environmental repercussions. However, with the benefit of hindsight and increased expertise, I am eager to implement a more advanced methodology in future experiments.
 

6. Did working on this project give you any ideas for other projects? 

While I was collecting the microbiome samples from my local river, I pondered about the areas directly exposed to Lithium Ion Batteries. This led me to investigate EV battery production plants and E-waste sites as potential locations for future in situ sampling. Having found numerous E-waste sites nearby, and  production plants  in Texas, California, Nevada, and other such locations. In order to further my research,I'm currently reaching out to these facilities to request samples and scheduling appointments with E-waste sites for microbial sampling


The outcomes of this research have the potential to shed light on safety standards and draw attention to environmental implications of EV battery production and recycling, prompting discussion on regulating the process to mitigate environmental impacts. Similar to the research that led EPA's push for lead standards in the 1970s-1980s, I believe research like mine is essential for our planet's safety amidst the rapid expansion of the EV industry.


Additionally, my investigations into battery pollution revealed frequent fires in landfills caused by Lithium Ion batteries, prompting consideration of the environmental impacts of the byproducts from these fires. Furthermore, due to the potential for lithium batteries to release more heat compared to traditional fossil fuels while burning, I'm interested in exploring potential changes in burn injuries and first responders' actions during accidents. These are aspects I aim to pursue in future research endeavors.

 

7. How did COVID-19 affect the completion of your project?

There was no impact from COVID-19 to the completion of my project.