Week 9: Plastic 3 and the The Tunnel of Oppression

(mold on plastic 3)

Our plastic 3 is finally decontaminated after two PIA plate test showed possible mold growth. The final fix was boiling the alcohol treated plastic directly in the hood. Now tomorrow we can begin to co-incubate plastic 3 with the Pseudomonas and the TSB. Also tomorrow we are going to check the 7 day samples and possibly remove the plastics from the Pseudomonas. Completely unrelated but very interesting; I attended the tunnel of oppression here on campus yesterday. It was very emotional, eye opening , in your face, and informative.  I recommenced everyone go out and see it because it is important and the different organizations have done an amazing job of conveying the challenges they and many others are facing. It is more then worth your time. I took my kids and they had so many questions and I hope that it will be that way for many people, It should provoke many questions, thoughts, and feelings. Go check it out next spring!

Week 8: Phase 2

This week has mostly been prep for phase two of our project; Will the Pseudomonas degrade micro plastics 3, 4, 5, and 6? We have all our plastics sterilized and have our bacteria ready to co-incubate tomorrow. (might not use plastic 6) So it been a relatively slow week which is amazing because it has given me the chance to join the Robotics Club, something that I have wanted to do since finding out we had a Robotics club at PC. I am so excited for Phase two and also to learn how to build a robot and to also program it. Lots of fun and exciting stuff ahead! 
A very broken and incomplete Roomba (photo shown above) will be the chassis for my "Ratbot"

Week 7: WAESO was great

After all the fear and anticipation the WAESO conference was actually really nice and I am so glad we went. It was not as big as i thought it would be and honestly our posters looked amazing! Just how the poster is printed out makes a huge difference. Ours stood out. We are not sure if any of our poster have been recognized,  we are waiting for an email.  Even if they are not, we definitely leaned alot so the experience alone was well worth it.  The guest speaker were incredibly inspiring and now I'm actually excited to go to the next conference.  Once again,  shout out to my wonderful team, Ibrahim, Kassy, and Dr. Cotter! Job well done!

Week 6: The day before the WAESO conference

As you can see this image shows a whole other conference. So here we are, on the eve of my first scientific conference ever and I just finished applying for the next one at ASU west. I almost have no words. I am excited and nervous for the conference tomorrow but the thought of another one around the corner kind of kills the vibe. I am going to soak in tomorrow as best as I can and take my experience with me to all the other conferences coming up. I am sure by the next two I will feel like a pro (I hope). I cant wait to fill everyone in on how its goes. Lastly, but most importantly, I need to say that my team, Ibrahim and Kassy, have done such a great job on these posters. Working around so many different schedules and still getting it done on time is absolutely amazing. You guys should be proud of your hard work and I know that it will show tomorrow. I am very honored to be a part of this team. And I can not forget Dr. Cotter for always being there to help, guide, and push us when ever needed. She is an extraordinary Mentor and leader. Thank you guys for all you help so far this semester. Good luck tomorrow.

Week 5: BUSY BUSY BUSY

This week for s-stem I have been doing my maintenance duty. The first day was pretty hard, only because I had to get rid of all the old bacteria tubes.  The smell was about ten times worse then the ugly smell of the incubators (barf!). I had to separate the glass tubes from the plastic caps,  the caps go to the autoclave and the glass tube is disposed of in the proper glass bin.  The dead bacteria is simply poured down the drain. My second task was to put all the clean glassware back on the shelf, a much more pleasant task.  The past couple weeks have been very very busy with WAESO as my team preps for the upcoming conference.  So so much to do.

WAESO Conference Abstract

It was recently discovered that drinking water all around the world contains microplastics that have made their way from wastewater into the environment and back to drinking water treatment plants. Microplastics are any piece of plastic smaller than 5 mm. They are small enough that they do not get filtered out during the water treatment process so they end up in our water supply and can continue back on a cycle of contamination. Microplastics are found in 94% of the tap water in the United States. The human and ecosystem health implications of this are not fully known however this is an emerging issue that will most likely require innovation and the design of strategies to reduce the amount of  microplastics contamination in our recreational and drinking water supply. One proposed solution is the use of microbes to aid in the biodegradation of microplastics. Several studies have shown that Pseudomonas putida and Pseudomonas fluorescens have been found to grow on polyvinyl chloride (PVC), while Pseudomonas stutzeri has been found to grow on hydroxybutyrate. Studies have shown that, in addition to colonizing microplastics, Pseudomonas species may play a role in the decomposition of plastics such as polyvinyl alcohol (PVA), polypropylene and polythene. This raised the question of, what types of microplastics can different Pseudomonas species colonize and biodegrade? The purpose of this project was to determine if Pseudomonas stutzeri along with other Pseudomonas species (Pseudomonas aeruginosa, Pseudomonas putida, and Pseudomonas fluorescens) are capable of colonizing plastic types PVC (plastic type 3), Low-density Polyethylene (LPDE) (plastic type 4), Polypropylene (PP) (plastic type 5), and Polystyrene (plastic type 6). The initial goal of this project was to establish protocols for the colonization, isolation, and genetic identification of the Pseudomonas species from each of the four microplastics tested.

To achieve this goal,  Pseudomonas stutzeri was grown on Pseudomonas Isolation Agar (PIA) and Pseudomonas fluorescens (PF) agar to determine the characteristic colony morphology. ribosomal-16S primers were also used to identify the genetic relationships between P. stutzeri, P. aeruginosa, P. putida, and P. fluorescens. SEM was used to image the microplastics that were co-incubated with the different Pseudomonas species at 20x, 620x, 1150x, 4400x, and 7500x. Future studies will be used to determine which Pseudomonas species can aid in the biodegradation of microplastics PVC 3, LPDE 4, PP 5, and PS 6 .

Pseudomonas Stutzeri and Micro Plastics

            This research project was already in progress a semester before I joined in. When Dr. Cotter first explained the research to me I was very excited to get the chance to aid in such an amazing and necessary project. The significance of this topic is huge globally. Micro plastics are found in almost every environment on the planet. What are micro plastics? Plastic debris can come in all shapes and sizes, but those that are less than five millimeters in length (or about the size of a sesame seed) are called “microplastics.” (NOAA) It is critical that we know about micro plastics and how they affect the ecosystem. It should be obvious that microplastics are not a natural part of our diets, so it should be alarming when we see statistics like this one from a Nature.com article titled River plastic emissions to the world’s oceans stating, “We estimate that between 1.15 and 2.41 million tonnes of plastic waste currently enters the ocean every year from rivers…” (Lebreton, L.C.M. et al) How do we begin to solve this global issue?

            Enter the Pseudomonas. “Pseudomonas stutzeri is a gram-negative bacterium… The presence of P. stutzeri in virtually all environments has led to it being called "almost universal." Soil and marine waters are two environments where P. stutzeri can be found.” (Baughman, W. et al) Pseudomonas stutzeri’s abundant presence in the environment makes it the perfect candidate to possibly colonize and degrade microplastics. Several studies have already investigated microbial degradation of plastics with success.

            My research question is: Can Pseudomonas stutzeri colonize micro plastics 3, 4, 5, and 6? Research will be done through experiments and observation.

Name	I/D/C	Symbol	Units	Description
Plastics 3,4,5, & 6	Independent	-	-	PVC, LDPE, PP, & PS-E
The colonization of microplastics	Dependent	-	-	Positive=growth Negative=no growth
Inoculating TSA with microplastics for evidence of Pseudomonas growth	Control	-	-	Growth either on microplastics or TSA.

            My hypothesis is that P. stutzeri will colonize micro plastics 3, 4, 5, & 6. At the end of my experiments I may find that P. stutzeri will not colonize micro plastics 3, 4, 5, & 6 and that may bring on further experimentation with other microplastics in the future.

Works cited

 

Baughman, W., Balaze, K., and Bruce, A., students of Prof. Jay Lennon at Michigan State University. (24 April 2012) Pseudomonas stutzeri. 8 February 2018. https://microbewiki.kenyon.edu/index.php/Pseudomonas_stutzeri

NOAA. (10 October 2017).  What are Microplastics? 8 February 2018. https://oceanservice.noaa.gov/facts/microplastics.html

Lebreton, L.C.M., Van der Zwet, J., Damsteeg, JW., Slat, B., Andrady, A., Reisser, J. (07 June 2017). River plastic emissions to the world’s oceans. 8 February 2018. http://www.nature.com.ezproxy.pc.maricopa.edu/articles/ncomms15611