Final Report

Our work involves the utilization of Agar to incorporate and fuse with plants essentials oils in varying concentrations. Agar as a gel is porous and can be used to measure microorganism motility and mobility. The gel’s porosity is directly related to the concentration of agarose in the medium resulting in various levels of effective viscosity. Throughout the project 30 essential oils were used in three varying concentrations of 100uL, 200uL, or 300 uL or 500uL, 75uL, and 1 mL. This made for 90 samples that were tested against S. aureus, and 52 samples were proved to have antimicrobial activity against this species. Then 9 samples were proved to have inclusive results because they didn’t diffuse into the agar or because in the incubator it was disintegrating and 29 samples did not work at all. All of these surfaces that were proven to work against S. aureus have shown efficient antimicrobial effects.

As of now, I have 52 samples that have been proved against S. aureus. Some of the future works involve finding new essential oils, which have not been heard about in the United States. The research plan is to continue making antimicrobial surfaces and not only testing it against S. aureus which is a known skin infection but to try this against other species like Candida albicans. In further projects plans, I tend to make these antimicrobial surfaces into wound dressings or bandages that can be used in the military or medical field.

Through this whole academic year, I have learned that my passion is truly being in the lab setting. I enjoyed every aspect of research. It wasn’t always great results, but this is all a learning process. Research was what my schedule revolved around each day. I am so happy with all that I have accomplished with this project! All the times in the lab and learning about each oil and the benefits that it has for you was the most interesting part. When I saw the positive results, that the oil would inhibit all the growth of the bacteria, I would get so excited that it worked! After doing research, I realized that I could pursue an MD/PhD or get my MD and work on the side doing research, any of these choices would make me happy. I enjoyed research and learning about everything that has to go with it. I am very thankful for my mentor as we made sure the project was presented at different conferences to gain feedback from people.

I presented my research project at the Annual Meeting of the Society of Fellows at Pace University and handed in a manuscript of my project. It won the Excellence in Science Award and I was very happy. Also, I attended a National Conference Meeting in San Francisco for the American Chemical Society this month and came back with excellent feedback and great connections. It amazes me how much of an impact this project has on others. It is always exciting to interact with others and engage and learn from them new techniques. The most exciting thing that I got when I was at the National Meeting was someone coming to my poster and after I presented to them, they said to me, “ This field is meant for you and I don’t often say that! This work will have a huge impact in a period of within five years! You are up to big things kid! Congrats!”

In general, this wouldn’t have been possible without my mentor, Dr. JaimeLee Rizzo. Thanks to my mentor for her guidance and advice has gotten me this far with this project! We expect that this project will continue to foster bigger things! I am very thankful to have been chosen to the program! It was a remarkable experience!

Blog #3

Previously before all of the samples, from the mid-year progress report, showed that inhibition worked on the surface. That all the samples were proven to work efficiently against the bacterium tested, S. aureus. My mentor and I decided to send it to another laboratory for verification of the results. Dr. Melkonian, a biologist, conducted the same experiment on the agar samples that have proven to be efficient against S. aureus. In fact, when she conducted the experiment, she ran two different methods which were the visual growth, the same method the student performs, and the subculture growth. She realized that some of the essential oils killed the bacterium, while others ones inhibited the growth of the bacterium. As of now, the essential oils rosemary, neroli, bergamot, sage, red thyme, lemongrass, clove, thyme, and palmarosa in the highest concentrations of 1 mL showed that S. aureus is completed killed. There are no signs of inhibitions instead the bacterium is killed completely. For verification, the subculture was conducted, which is a removed loopful of agar from a previously streaked plate and incubating it in 2 mL Tryptic Soy Broth (growth media) for 24 hours, which all these essential oils killed the bacterium. The inhibition was seen in the oils that had the smallest concentrations, for example, all these oils varied from 100 uL-500 uL in the lowest concentrations and they all proved in those ranges to inhibit the bacterium only. It is concluded from the first round of testing that these essential oils have antimicrobial properties. We are currently sending more samples for verification of the killing or inhibition of the bacterium.

At this moment, I am trying to find new antimicrobial essential oils to infuse into the agar. In the lab now, I currently have chamomile to test and am in the midst of researching more essential oils. Thus far, we have 52 samples that have been proved against S. aureus and are sending them over to the laboratory for verification. Some of the works involve finding new essential oils, which are exotic to the United States. The research plan is to continue making antimicrobial surfaces and not only testing it against S. aureus which is a known skin infection but to try this against other species like Candida albicans.

Till this day, I am always mind-blown what an essential oil can possibly due to saving your skin. People wouldn’t think that a surface that contains the essential oil could possibly do anything. If anything, people would suggest it has a good smell, but they don’t know that this oil can potentially save them in a skin infection. In fact, the more positive results that we are obtaining and more verifications of killing the bacterium which is fabulous news is mind-blowing. I would like to continue this project in the future for the creation of a wound dressing or bandage.

Mid-Year Progress

Since, the last time I wrote on this blog, I have conducted five trials which total up to 30 essential oils that were tested. Some of the questions that I raised throughout my research was why the infusion of some oils was not diffusing properly in the agar. So, I have tried a new method that is proving to work. The method has been applied to trial 3 and so on. Trial 1 and 2 are not exposed to this new protocol. The protocol to make the agar is to do the following: obtain a 500 mL Beaker to make the agar, measure out 250 mL of Distilled Water, add 2.50 grams of Tryptone, add 1.25 grams of Yeast Extract, add 2.50 grams of Sodium Chloride (NaCl), and add 3.75 grams of Agar. Once all the materials have been added to the beaker, stir the beaker for a few minutes and place on the hot plate for approximately 15-20 minutes, until it boils. Then the new protocol is: when pouring into the plates, take a falcon tube and fill it with 10 mL of agar. Then add the measured amount of essential oil and shake gently. After you shook it gently pour out onto the plates for a more uniformed surface.

Trial One: I have concluded that out of the six essential oils that were tested four of them are efficient against the bacterium S. aureus. The four essential oils that showed good results include sage, red thyme, ginger, and lemongrass. The four essential oils show that as the concentration increases from 500 uL to 750 uL to 1 mL, the results to inhibiting the growth of the bacteria has also increased. The other two oils, carrot seed, and myrrh, showed no efficient results at all. In fact, those essential oils looked almost identical to the agar blank, therefore I can conclude from trial one that carrot seed and myrrh do not have antimicrobial properties.

Trial Two: I have concluded that out of the six essential oils that were tested two of them are efficient against the bacterium S. aureus. The two essential oils that showed good results include thyme and clove. Propolis was the only sample that did not diffuse entirely into the agar. This led to inaccurate readings since on the sides there were bacteria, but on the oil there was none. The two essential oils show that as the concentration increases from 100 uL to 200 uL to 300 uL, the results to inhibiting the growth of the bacteria has also increased. The other three oils, Guava Seed, Neem Seed, and Frankincense, showed no efficient results at all.

Trial Three: I have concluded that out of the six essential oils that were tested two of them are efficient against the bacterium S. aureus. The two essential oils that showed good results include Neroli and the combination of clove and frankincense. This is the first time a combination was used because previously, it was shown that Frankincense was not efficient at all, in fact as a large amount of it was used with a small amount of clove, which has strong antimicrobial properties, the testing proved to have no growth of bacteria at all. More combinations will be conducted in the next round. All samples were diffused perfectly into each other because the new protocol method of the falcon tube was applied. This protocol of using the flacon tube made the surfaces uniform. The other four oils, Yarrow, Black Elderberry, Argan, and Jamaican Black Castor Oil, showed no efficient results at all.

Trial Four: I have concluded that out of the six essential oils that were tested five of them are efficient against the bacterium S. aureus. The five essential oils that showed good results include Spearmint, Geranium, Palmarosa, Bergamot, and Rosemary. All samples were diffused perfectly into each other because the new protocol method of the falcon tube was applied. This protocol of using the flacon tube made the surfaces uniform. The only oil that did not show results was Wintergreen. One observation about this oil is that the color is pink. This is different compared to the other oils which are normally yellow and clear. The plates for Wintergreen were done with the same concentrations as the others which are: 500uL, 750uL, and 1 mL. The observation seen was that the essential oil was eating through the agar and also through the plastic. For the 500 uL, the plate looks like it is about to shatter, as for the 750 uL the plate shatters in the center, and for the 1 mL the plate shattered completely in the center. This only means that the results are inclusive because nothing can be recorded about the antimicrobial activity.

Trial Five: I have concluded that out of the six essential oils that were tested five of them are efficient against the bacterium S. aureus. The five essential oils that showed good results include Peppermint, Oregano, Rosehip, Keylime, and Eucalyptus. All samples were diffused perfectly into each other because the new protocol method of the falcon tube was applied. This protocol of using the flacon tube made the surfaces uniform. The only oil that did not show results was Wintergreen. The plates for Wintergreen were done with the same concentrations as the others which are: 100uL, 200uL, and 300uL. After trial four was conducted and the results were seen that the samples were disintegrating meant that I had to lower the concentration. In fact, when decreasing the concentration, no inhibition occurred and also in all 3 plates, there were cracks coming through. Therefore, I can conclude that wintergreen is made of very strong components and cannot be tested in the incubator for an antimicrobial surface.

Background to Project: Infusion of Native Oils To Synthesize Antimicrobial Properties

Infusion of Native Oils to Synthesize Antimicrobial Surfaces is the project I will be working on this academic year. To further explain, the meaning behind the project I have provided background information as well as goals and materials to reach for successful results.

The challenge to maintain a sterile environment and protect patients in a clinical setting has grown in the recent years, due to the exposure of microorganisms. The discovery of the antimicrobial surfaces in previous research has shown a minimized growth in microorganisms like: bacteria, fungi, viruses, etc. Challenges still arise in creating surfaces because of the difficulty to industrialize, the non-uniformity throughout the surface, and the activity of the antimicrobial agent being wiped off.

Our work involves the utilization of Agar to incorporate and fuse with plants essentials oils in varying concentrations. Agar is a polymer that is composed of subunits from the sugar, galactose. Agar surfaces are not degraded or eaten by bacteria and also serves as a firmer and stronger surface. Agar, as a gel, is porous and can be used to measure microorganism motility and mobility. The gel’s porosity is directly related to the concentration of agarose in the medium resulting in various levels of effective viscosity. The agar surfaces are infused with some antimicrobial oils such as: Propolis, Neem Seed Oil, Black Elderberry, Yarrow, Tamanu, Rosehip, Ginger, Sage, Argan, Guava Seed, Myrrh, Frankincense, and Neroli, Red Thyme, Lemongrass, just to name a few.

All surfaces are to be tested against the gram positive bacteria strain S. aureus. S. aureus, a gram-positive bacterium is the leading cause of skin and soft tissue infections in humans. It was designated to be the most important bacteria that caused diseases in humans. Annually, it was estimated that 500,000 patients in the United States were affected by this bacterium in clinical and hospital settings, that even some of the strains weren’t resistant to the antibiotics. Previous studies and research has shown that some of the strains have developed resistance to antibiotics, but some of the strains aren’t killed completely, which only causes the bacterium to multiply, cause the infection, and eventually lead to a more serious condition to be treated.                        

Therefore, the creation of the antimicrobial surfaces can minimize the growth of microorganisms like S. aureus. The surfaces are made from agar, which previously mentioned is known to maintain a firm and strong surface. The creation of having a firm and strong surface is to perhaps aid as a wound dressing or bandage in the medical and military field. The surfaces are encoded with the native plant essential oils, which are all known to have strong antimicrobial effects. The essential oils are added onto each surface in various concentrations, which will then be tested against S. aureus to see if the essential oil properties can inhibit the growth of this organism.

The discovery to acquiring new data since this project started is important because if surfaces that are being made naturally are efficient, they can be considered for wound dressings and the experiment then will change gears to testing how pure the surfaces are and even go further into testing it on humans. The ability to perhaps think that the antimicrobial properties of these essential oils can potentially kill the bacterium, S. aureus, would lead to a huge platform in creating bandages for the military and medical field.