MARIN ACADEMY RESEARCH COLLABORATIVE​
STUDENT BLOGS
ADDITION OF ANTI-BACTERIALS TO FLUID THIN FILM COATINGS(FTFC) TO PREVENT MICROBIOLOGICALLY INFLUENCED CORROSION
My area of scientific interest is chemistry, specifically looking at corrosion prevention and protection in aeronautical and nautical applications. Currently, I plan on looking at the application of antibacterials to traditional corrosion inhibitors as a way to reduce microbiologically influenced corrosion. Traditional forms of prevention rely on creating coatings which resist colonization (1), however, I plan on looking at the role active inhibitors could play. ​
1. P.M. Natishan, D.C. Hansen, G.I. Loeb(1996). Inhibition of Microbiologically Influenced Corrosion. http://www.dtic.mil/docs/citations/ADA305227
Making media and cutting steel
March 20th
I have been cutting all of my steel to length which has worked well. Today I will finish doing that and treating the steel so that it is clean. This involves dousing it with HCl and then NaOH. Then I will autoclave so that I am ready to weigh it. Once I actually put it in the tubes it will be sealed, so that shouldn't be a problem. From there I will go to LBL to add the bacteria, which then will hopefully eat the steel. The main thing I need to work on moving forward is my actual timeline so that I finish in time. Three reagents are still on their way, but once they arrive I will be able to make the media. Overall I am very excited about the state of my project. Unrelated but today I interviewed two potential sophomores so that was fun.
Going to LBL
March 13th
I am going to Lawrence Berkeley next week to innoculate the media and my Hungate tubes. Valentine has been a huge help in offering to let me do this at her lab. The carbon steel arrived today, so I am cutting it and treating it today. Additionally, I am starting to make up the media so I can put it in the Hungate tubes. We have ordered the Hungate tubes and they arrive this Friday. Overall I am super excited for the next step in my project and to get all of my data. I will have all of my data before spring break which will give me a good chance to get a jump on the analysis so that I am finished in time for the end of the year.
Starting to test
March 6th
I contacted Valentine about the oxygen resistance of DvH and she gave me a couple of possibilities of avenues for how I could test. She said that the DvH is fairly resistant to oxygen, but only once it's in an established colony and has started to produce sulfide. If the colony is not established, exposure to too much oxygen will kill them or severely slow down the propagation of the colony. If I use Hungate tubes, she suggested bubbling nitrogen through them to saturate them with a non-oxygen-containing gas. I would then tap the tubes to force out any remaining oxygen. From there I would thaw the glycerol stocks and innoculate with a syringe and needle. Additionally, I could use the method which she suggested where I open the tubes in the anaerobic chamber for as long as possible to force the oxygen out and let the medium saturate with nitrogen and carbon dioxide. The one problem with this is that I might let oxygen in when reopening the chamber to innoculate. The final option she presented me with is that she could provide me with the correct Hungate tubes, and then I could use those. I'm going to ask if it would be possible for me to innoculate the tubes at her lab and then bring them back to incubate which would be the best option.
Data Presentation
February 20th
My meeting last week with Valentine went great. She gave me two cultures of two strains of DvH each, as well as some tips on how to properly culture them. Now that I have the bacteria as well as the proper culture method I can get started with my testing soon. I do need to email her back about whether or not using Hungate tubes would be a viable method for the starter culture. I know they would work for the test cultures, I just need to figure out the best way to keep a starter culture going. Other than that, I'm looking forward to minicourse next week, as well as the rest of the year. After I finish my testing I should be able to write the rest of my manuscript fairly easily as my topic has not really changed, so I still have all of my initial research available and useful. I plan on first doing the corrosion test and then if I have time and am able to figure out methods, then doing a microtiter test plate. This is because the microtiter will be significantly harder to work with and culture properly in the anaerobic chamber we have. At the very least the main test should work, which is the more important one anyway. The bacterial growth is less important than whether or not the corrosion actually happens.
Data Presentation
February 6th
I just had my data presentation which went well. I need to look more into specific types of statistical tests so that I can properly analyze my data once I have it. Additionally, I have changed one aspect of my methods. I will now be using 96 well microtiter plates filled with Postgate medium with a dilution of the FTFC added to test antimicrobility. I will then perform a separate test with steel coupons in order to determine anticorrosion properties. To test corrosion I will perform both a visual analysis as well as weighing each coupon before in after. This will allow me to determine whether there is any corrosion in general, as well as what type of corrosion is actually occurring. I am still waiting for Lucy Zheng to get back to me, but I hope I will have the source by the end of this week. Other than that, I'm probably going to make the media soon, as well as practice the sterile anaerobic technique for the plating. Additionally, I need to see if I can purchase one-way valve tops for the media cultures so that I can bubble nitrogen in for the corrosion testing more easily. I need to also get more familiar with the anaerobic chamber. Other than that, I'm excited to get my project rolling, and I'll hopefully have the bacteria next week.
A search for Desulfovibrio
January 30th, 2018
I was able to get in contact with Dr. Walian at Lawrence Berkeley who subsequently put me in contact with their research associate Lucy Zheng. She informed me that at LBL they contract to an outside source who would be able to help me obtain Desulfovibrio Vulgaris in the form of frozen aliquots. She will be contacting them next week, and I await a response. As for this week, I have gotten a lot of work done on my intro and methods, as well as prep for my data presentation. I will mostly be checking in with the group about statistical analysis, as well as specific numbers for my control vs test samples. One thing that has worked out well in my favor is that our turbidity test kit uses 10mL vials which are the perfect size for a sample. I can culture each sample directly in test vials, thereby eliminating the need for working continuously under anaerobic conditions, I can just seal each vial instead. This will allow me to optimize my setup and perform many tests at once. Additionally, this will allow me to measure turbidity often with few repercussions on bacteria growth. I go into the month of February confident, as long as I am able to properly secure the bacteria. Hopefully, by next my post, I will not only have a completed methods but bacteria and media as well.
Emails for Bacteria
January 23rd, 2018
Since my last post, I have met with Stori regarding my materials list and we can properly obtain all of the chemicals, so that shouldn't be a problem. The only thing we can't reasonably get our hands on is the Desulfovibrio vulgaris itself. Currently, I have emailed Dr. Peter Walian and Dr. Gary Andersen from Lawrence Berkeley and I am awaiting their responses. I have spoken to Dr. Andersen before, and Dr. Walian has worked with Desulfovibrio vulgaris, so they both seem like they could help me. If they either don't get back to me or cannot supply me with bacteria then I will have to attempt to find other institution or organization that could supply me with the bacteria. Additionally, I will need to look into other potential bacteria to use, especially if either we or Dominican already have it. Before next week I need to identify another potential bacteria (or several), as well as look for other places which could potentially supply me with the D. vulgaris. And as always, I will continue to work on my intro and methods while I await responses and hopefully my materials. I am also beginning to prepare for our first data presentation.
Methods & Manuscripts
January 16th, 2018
At this point, I've created my action plan for January and have started working. I've compiled a list of chemicals and other material I need, as well as started working on my methodology. I have to meet with Stori to ensure that we can actually obtain all of the chemicals, but it seems like they should be fine. Additionally, I've delved further into my research for my introduction and have some good papers lined up now that I'm making my edits. Overall I'm still lined up to start my testing at the beginning of February and do my data analysis immediately after that. As I am just collecting one data point (turbidity), it shouldn't be too difficult. One thing I need to identify is how many controls I will be doing and of what types. I could use the same controls as I did for Science Symposium, although I'm not sure how helpful that would actually be. I also need to determine the actual amounts of FTFC to add. Furthermore, if the turbidity testing works, I can look into visual analysis for the corrosion testing. Overall, this week I just need to check in with Stori and get some good work done on my manuscript (especially the methods).
Back from Break: January
January 9th, 2018
In our first class back from break we checked in on the course guidelines and the general plan for the semester. I realized that in all likelihood there is no week in January where I will be able to come in to test the turbidity every single day, so I will instead focus on just getting the test completely set up. By the end of January, I hope to be completely ready for my tests, which I will start the second week of February, as well as be done with my intro and methods, with rough drafts of the parts of my discussion and results, at least the parts I can complete without data. This should put me on the path to finish my manuscript draft by the end of March, to hopefully complete my final draft by the beginning of May, or at least be close enough that there is only a small amount of editing left. Then, during May, as well as the summer, I can pursue publication in one of the secondary journals we have discussed in class. Additionally, I would like the opportunity to share my research with the individuals who have assisted me, such as Dr. Frost. Finally, this would put me on track to be ready for the wildcat colloquium. For the rest of this week, my work will mostly entail prep and solidification of my methods.
Senior Year Quarter 2 Update
December 11th, 2018
As it turns out the ASTM test is not going to be suitable for my needs. The trial is not only too complicated to build, but it would also be very difficult to modify. Additionally, to actually perform the test I would need to purchase several other ASTM standards putting the total cost far out of both budget and feasibility. That being said, I found another study with a modified sulfate-reducing bacteria culturing process that should work both for just culturing the bacteria and actually testing the antimicrobial properties of the FTFC. It uses a modified Postgate B Medium which is biologically free of iron. This means that for bacteria growth to occur a steel coupon has to be added. In addition to testing the actual amount of bacterial growth, I will also be able to perform corrosion analysis on the coupon itself. These two tests should give me all of the information I need to determine the efficacy of FTFC as a preventative measure against microbiologically influenced corrosion (in relation to sulfate-reducing bacteria).
I plan on starting off next semester with data collection. It should take 7 days, so I will most likely do a Wednesday to Wednesday test period, or something similar. Getting all of my testing done in January will set me up perfectly to focus on writing for the rest of the semester. If I can get everything written up in the first quarter, then I can pursue publishing and presenting and various organizations for the second quarter. Additionally, I can share my research with CorrosionX, as they may have an interest in the work I've done. Overall, it seems that if I can complete my data collection on time, my project will be on track to be completed by the end of the year.
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Ismail, Mardhiah & Yahaya, Nordin & Bakar, Akrima & Noor, Norhazilan. (2014). CULTIVATION OF SULPHATE REDUCING BACTERIA IN DIFFERENT MEDIA. Malaysian Journal of Civil Engineering. 26. 456-465.
Dr. Gary Andersen Quick Update
Novermber 7th, 2018
I finally was able to get in contact with Dr. Gary Andersen but it turns out that he is currently in Singapore, and as such will not be able to provide much help as of right now. Additionally, he said that he does not have very much experience with culturing sulfate-reducing bacteria specifically, although he said that it is helpful that we are set up for anaerobic growth. Once he gets back from Singapore he said he will ask some of his peers if they have any suggestions, and if so, he will put me in contact with them.
Additionally, I have identified ASTM D5969-11 as a potential test method for corrosion inhibition testing. It uses a synthetic seawater and sulfate-reducing bacteria commonly occur in anoxic seawater so it seems that it should not be too hard to adapt the standard for my purposes. Hopefully, both Dr. Andersen and the standard will pan out and help move my project forward.
Dr. Frost Meeting Update
October 20th, 2018
My meeting with Dr. Frost went very well. We discussed my problems with precision during science symposium, as well as my problems with how to actually perform the corrosion analysis. He had several suggestions and as always was extremely helpful. He suggested that I look into the American Standard Testing Materials standards as a potential way to perform the corrosion evaluation. The produce standards that are peer-reviewed and not only have given methodologies but also confirmed margins of error as well as p-values. This would allow me to very accurately determine the rate of corrosion on a piece of metal. I'm going to look more into which specific test would best fit our needs (he recommended D665.
He is also putting me in direct contact with Dr. Gary Andersen whom we had spoken about before to further help with my project. He is at Lawrence Berkely and an expert in microbiology. He should be able to help me figure out a better way to culture the bacteria. I hope to speak with him in the coming weeks about a good way culture them as well as test the antimicrobial properties with the culture.
SENIOR YEAR QUARTER ONE UPDATE
September 17, 2018
Since the end of junior year and science symposium, I have made a lot of progress with how I will actually be completing the final version of my project. I am now moving away from applied research and into the realm of basic research. I will be focusing on whether the fluid thin film coatings themselves have any antimicrobial properties and whether or not they can in any way prevent microbiologically influenced corrosion. The lack of general information on this topic is the biggest issue when trying to come up with solutions. Additionally, if I discover that the FTFC itself has antimicrobial properties that will erase the need to synthesize a new FTFC or add an antibiotic to an existing one.
The biggest step in my project I am currently working on is refining my methodology. Science symposium gave me interesting results but I cannot confirm their validity until I refine my methods and perform in vivo testing. My data will be useless unless I can successfully culture bacteria on steel in marine like conditions. I look forward to the coming year and completing my project.
SUMMER PLANS & CONTACTS
February 5, 2018
I am still waiting to here back from Naval Academy Summer Seminar and Air Force Academy Summer Seminar. These programs would each allow for me to participate in scientific research at two of the top STEM colleges in the country. If I can attend either of these programs it would help give me a better idea of what science looks like at the college level. Additionally, I am finishing up my application to once again teach Math and Science at Aim High, which would be helpful as it forces me to be able to convey the ideas of my project in simpler terms, something that would be incredibly useful for any presentations I may have to give. Another thing I am looking into is the various programs the Naval Research Laboratory offers to further extend the capabilities of my MARC project. This includes the Student Volunteer programs and SEAP. I have to get into contact with the organizer to determine whether or not I am eligible. I have learned that the Navy does not publish contact info for their scientists, but I have emailed their help-desk and am waiting for a response. I have also been making several updates to my research proposal.
RECENT MEETING
January 30, 2018
Last week I went to dominican and met with Dr. Frost, the Chemistry Professor. We talked about several aspects of my project, including relevant commercial products like BioBor(a boric acid derivative that functions as a broad spectrum bactericide in diesel)Â and how through studying their properties, I could apply similar products to my own studies. We also talked about the core ideas of corrosion prevention, focusing on the importance of the metal oxide layer. When researching and testing the bacteria, I will need to pay close attention to not only whether or not the bacteria induce or inhibit corrosion, but also whether or not the bacteria have any noticeable impact on the metal oxide layer. Additionally, he recommended that I focus on killing and inhibiting the growth of those bacteria which induce corrosion as it will have a universal positive outcome as opposed to focusing only inducing the growth of the bacteria that prevent corrosion as they will not grow everywhere or on every surface. Additionally, Dr. Frost gave me the contact information for Dr. Gary Andersen developer of the phylochip and an expert on microbiological systems to further guide me in my project.
MORE RESEARCH AND POTENTIAL MENTOR CONTACTS
January 22, 2018
Recently I have been going over a lot more research and learning more about how biofilms can change the electrochemical conditions surrounding and influencing corrosion. One of the interesting things I have learned is that not only can bacteria indue corrosion, they can also prevent it, which might be a further area of study. The potential for being able to seriously reduce corrosion just by applying a bacterial layer is huge. At this point, my research is definitely starting to come together, and I am at a point where I am starting to understand the topic I have chosen.
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Although I have still not been able to get into contact with anyone from the CCSE, I do have a meeting scheduled with Dr. Frost from dominican on this coming Wednesday(1/24). I will be visiting him in his classroom and going over my project and potential ideas to make it more tangible and feasible.
FUTURE RESEARCH AND SEMESTER PLAN
January 11, 2018
The most recent development was the decision to abandon the project centered around active carbon sequestration through the use of molten carbonate fuel cells and instead focus solely on corrosion prevention and treatment. This decision came after a phone call with Dr. Matt Lucas from the Center for Carbon Removal discussing the potential risks and pitfalls of working with molten carbonate fuel cells. With my current knowledge and skills, a project centered around corrosion is not only safer, but much more feasible than one using molten carbonate fuel cells.
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Moving forward this means that I have to invest a large amount of time reading more literature surrounding my field of interest, as much of my old research is now inapplicable. Specifically, I want to learn more about the work being done at the Center for Corrosion Science & Engineering of the U.S. Naval Research Laboratory as the work being done is both relevant and readily available. Furthermore, I plan on contacting experts in this field(including those working at the CCSE) both for general guidance and information, as well as for potential mentoring opportunities.