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Effectiveness of Lauric Acid and Adrographis Extract on Skin Pathogens

This experiment served to find a natural alternative to antibiotics against skin viruses, and to prevent development of superbugs. The experiment was conducted at the University of Florida Healthís Microbiology Lab under supervision of Dr. Yvette McCarter. The items tested were andrographis extract, a bitter plant extract, and Lauric acid, the main fatty acid chain with antibacterial properties in coconut oil. The organisms that were tested on were Candida albicans, Staphylococcus epidermis, and Staphylococcus aureus. 
     One of the biggest medical problems today in developed countries is the overuse of antibiotics. This overuse is causing super bugs, such as MRSA, Methicillin Resistant Staph Aureus, to gain resistance to antibiotics making it a superbug. As many as 100,000 US hospital Patients die to antibiotic resistant superbugs every year. Doctors are taking steps to defeat the spread of these super bugs. They are trying to accomplish preventing the spread of bacteria between patients, infections related to surgery and improve antibiotic use. Some doctors are searching for natural alternatives to help prevent superbugs. One of the most well known natural bacteria fighting substances is coconut oil.

     Researchers have known about the beneficial properties of coconut oil over antibiotics since the 1960ís and recently, this has been documented in publications all around the world. The main fatty acid found in coconut oil is Lauric Acid, which is a medium chain fatty acid which can also be formed into monolaurin, an antiviral, antibacterial, and antiprotozoal monoglyceride, this is used in the human body to destroy viruses such as HIV, herpes, influenza, most pathogenic bacteria, and can also be used as topical antibiotic. The benefits of coconut oil surpass the the benefits of antibiotics not only because of their widespread actions, but also because of its affordability, and natural availability. Among natural ingredients in our food, Coconut Oil has wide spread antibacterial, antiviral, and topical including superbug fighting properties that consumers can incorporate into their diet daily.
      Lauric acid has many major benefits for fighting acne and other infections. The first one is itsí strong antimicrobial and antiviral properties. Some studies have shown that humans can take advantage of the immune enhancing effects that lauric acid possess. Another benefit is that Lauric Acid helps fight antibiotic resistance. Many doctors agree that coconut oil should be used instead of standard antibiotics to treat infection because the build up of antibiotics causes resistance. The last benefit is that it benefits skin health and can naturally fight acne. 

     Andrographis is known as the ďKing of bittersĒ, and has been used in India for hundreds of years. Along with anti-bacterial properties, andrographis is also an antidote to scorpion venom. It is mainly supposed to help fight against influenza and upper-respiratory diseases. The main reason why Andrographis is a better choice than antibiotics, is because along with killing bad germs, it leaves good germs unharmed. 

     The three organisms tested on were Staphylococcus aureus, Candida albicans, and Staphylococcus epidermis. Staphylococcus aureus is one of the most important bacteria that cause diseases in humans, and the leading cause of skin and other soft tissue diseases. It can also cause blood stream infections, pneumonia, and bone and joint infections. Staph is most commonly spread by contaminated hands especially in Immunocompromised people. Most Staphylococcus aureus infections are hospitalized. Although Staphylococcus aureus can be treated with intravenous antibiotics, too much of those antibiotics can transform Staph into MRSA, the superbug which feeds off of antibiotic resistance and encourages the medical industry to look to natural alternatives that are universally published to cure this disease. 
     Candida albicans is a strain of skin infecting fungus. Even in normal conditions, skin does contain little amounts of Candida albicans. Some types of Candida albicans infection are, athletes foot, oral thrush, and vaginal yeast infection. The treatment for this organism is topical lotions or creams. Staphylococcus epidermis is also a skin infecting bacteria. They usually colonize in mammals and humans. It differs from Staphylococcus aureus by lacking the enzyme coagulase.
 
     Staphylococcus Epidermis consists of forty gram-positive bacteria. The amount of infections that Staphylococcus epidermis is causing is starting to rise leaving doctors to find a better cure than antibiotics. Although Staphylococcus epidermis does not cause many health problems, it can cause serious complications leading to death, making it a much stronger bacteria than itís relative, Staphylococcus Aureus. The majority of people who carry this disease usually have some kind of prosthetic in side them such as artificial heart valves and catheters. Due to the fact that the human body always contains an amount of Staphylococcus Epidermis, what causes the disease is when extra bacteria seeps into your skin creating a biofilm which keeps the bacteriaís metabolism low and doesnít allow antibiotics to enter, causing severe complications. The only way to treat this bacteria is vancomycin,a glycopeptide antibiotic. The side effects of this drug include nausea, hearing loss, diarrhea, and kidney problems.

     The procedure went as followed. One blood Agar plate was taken out, then a plastic swab was used to swab the bacteria, and then doused into a one hundred percent saline solution which was then compared to a 0.5 standard barium sulfate solution to make sure the clarity was equal to ensure that the colonies were not heavily populated or underpopulated. Then the whole process was repeated for the next two bacteria. After making sure the clarity was equal, the suspension was put on a cotton swab and was inoculated onto the agar plate. Then forceps were used to separate cotton discs .5 mm in diameter. Then a plunger was used to pick up 50 micro-liters of andrographis extract, and was placed on each disc. The Lauric acid was suspended in water to be dissolved, the solution was fifty percent concentration of lauric acid. Then the plunger picked up fifty milliliters of the lauric acid solution and was placed on five discs per bacteria, a total of fifteen discs were made of the lauric acid solution, and the andrographis extract. Then, the plunger picked up twenty-five milliliters of each compound, the lauric acid solution, and the andrographis extract, and a mix of the two were put on each one of the five discs per bacteria. Then the discs were transferred to each inoculated agar plate. Along with these two compounds and the mix, a control plate was made for each bacteria, using water, and a standard antibiotic disc , was used to compare the growth of bacteria. Then the plates went into the incubator set at 25 degrees Celsius for 24 hours and read immediately. Each step was documented in the composition book along with the results. 

     For this project, the hypothesis was ďThe mix of both, lauric acid, and andrographis extract will have the greatest zone of inhibition per each bacterium.Ē This hypothesis was chosen due to the inference that combined anti-bacterial properties will cause more inhibition than each one separate. The results were as followed, the five lauric acid discís zone of inhibition against Candida albicans were, 6 millimeters,6 millimeters,6 millimeters,6 millimeters,6 millimeters. The average being 6 millimeters. The five andrographis discís zone of inhibition against Candida albicans were 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters. The average being 6 millimeters. The five water discís zone of inhibition against Candida albicans were 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters,.The average being 6 millimeters. The five antibiotic discís zone of inhibition were 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters. The average being 6 millimeters. The five mix of andrographis and lauric acid discís zone of inhibition were 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters. The average was 6 millimeters. This data proves that nothing worked against Candida albicans. The antibiotic and water were sure to not work because this antibiotic did not have anti fungal properties. The lauric acid did not have a significant impact against it either, leading to believe that a higher concentration would make a difference. The Andrographis also did not have an impact on Candida albicans. The results of the mix proved that that even the combination of the two compounds were not powerful enough for Candida albicans. The results for the five lauric acid discís zone of inhibition against Staphylococcus aureus were 8 millimeters, 8 millimeters, 8 millimeters, 8 millimeters, 8 millimeters. The average was 8 millimeters. The five andrographis discís zone of inhibition against Staphylococcus aureus were 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters. The average being 6 millimeters. The five water discís zone of inhibition against Staphylococcus aureus were 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters. The average being 6 millimeters. The five antibiotic discís zone of inhibition against Staphylococcus aureus were 26 millimeters, 25 millimeters, 25 millimeters, 27 millimeters, 24 millimeters. The average being 25.4 millimeters. The five mix of andrographis and lauric acid discís zone of inhibition against Staphylococcus aureus was 7 millimeters, 8 millimeters, 7 millimeters, 8 millimeters, 9 millimeters. The average was 7.8 millimeters. The Lauric Acid, and the mix did inhibit the bacteria one-third of what the antibiotic could do. Andrographis still had no results. The results for the five lauric acid discís zone of inhibition against Staphylococcus epidermis were 11 millimeters, 12 millimeters, 10 millimeters, 11 millimeters, 11 millimeters. The average was 11 millimeters. The five andrographis discís zone of inhibition against Staphylococcus epidermis were 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters. The average being 6 millimeters. The five water discís zone of inhibition 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters, 6 millimeters. The average being 6 millimeters. The five antibiotic discís zone of inhibition against Staphylococcus epidermis were 30 millimeters, 32 millimeters, 30 millimeters, 29 millimeters, 30 millimeters. The average being 30.2. The five mix of andrographis and lauric acid discís zone of inhibition against Staphylococcus epidermis was 10 millimeters, 11 millimeters, 12 millimeters, 11 millimeters, 11 millimeters. The average was 11 millimeters. Lauric acid had a very large zone of inhibition compare to the tests, almost one-third of the antibiotic against Staphylococcus epidermis. 

     This project concluded that, Andrographis extract does not have anti-bacterial properties on skin bacteria, and lauric acid does. Andrographis, which may have upper-respiratory diseases fighting properties, did not contribute in the prevention of skin bacteria growth. Lauric acid proved a worthy competitor of the antibiotic in this experiment clearing near one-third of the antibiotics zone of inhibition on two of the bacteria, and neither had an effect on Candida albicans. A way to use this in daily life would be, topical creams with lauric acid to be applied to help prevent Staphylococcus Epidermis or Staphylococcus aureus. Since little cultures of Staphylococcus epidermis are found in your body at all times, using lauric acid creams will help to prevent any excess culture growing which can cause disease. Another way to use Lauric acid is to coat your skin in it making sure that no excess staphylococcus epidermis can come through causing a biofilm, a polysaccharide intercellular adhesion which keeps bacteriaís metabolism at an all time low and also makes antibiotics impenetrable, to form.
     During this project, there were many errors. The first trial, coconut oil was tested instead of lauric acid, and no results were found. The second time, when transferring fifty milliliters of lauric acid onto the five mixed discs, one was skipped over. When the plate was read, only four out of the five inhibited the cultures because the fifth one never obtained the compound. Further experimental ideas include, the variation of concentration levels on the same bacteria, testing lauric acid on more pathogen virusís to see if the results are the same, and developing a topical cream to help prevent Staphylococcus epidermis from forming in the human body. The most applicable way of using this study would be to coat the prosthetics inside people such as artificial heart valves, and catheters in lauric acid to prevent Staphylococcus Epidermis create a biofilm which would lower the bacteriaís metabolism and cause antibiotics to be impenetrable. With lauric acid covering these prosthetics, Staphylococcus Epidermis growth would be able to be inhibited and prevent that biofilm from forming.
Article By: Yash Gulani
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