Abstract

In this experiment we tested the capability of two substances, Allium sativum (common garlic) and Melaleuca spp. (tea tree) essential oil against a control and whether they effectively inhibited the growth of bacteria, specifically Kocuria rhizophilia. A bacterial lawn was created using agar petri dishes and live bacterial samples which then had the two test samples and the control sample applied to equal areas on two different petri dishes. The petri dishes were then left to incubate for a week before the resulting data was collected. Both samples displayed significant zones of inhibition where the control showed none. Melaleuca spp. essential oil and Allium sativum both express antibiotic properties which are active against Kocuria rhizophilia.

Introduction

With the rise of bacteriology in the 20th century, it has come to the public's attention that there are microorganisms which may cause different diseases and ailments depending if they are pathogenic or not. Since the discovery of this microscopic universe scientists have been working diligently on finding different compounds which exhibit inhibitory effects on these pathogens called antibiotics. However, could this all have been in vain? In the race to defend the human biome from these newly discovered foreign invaders doctors began prescribing antibiotic medicines for every thing, even viruses. Antibiotic compounds of pharmaceutical strength should only be used for chronic bacterial infections. According to the Family Journal of Health, “Antibiotic use—whether appropriate or not—has
been linked to rising rates of antimicrobial resistance, disruption of the gut microbiome leading to Clostridium difficile infections, allergic reactions, and increased health care costs”(2017). The use of antibiotic pharmaceuticals clearly needs to be reevaluated. Antibiotic pharmaceutical medicines are nonspecific and do not target a single bacteria, rather the entire microbiome becomes affected and the balance of gut flora is significantly impacted (Sullivan 2001). Our microbiome is our first line defense against pathogenic microbes attempting to damage our internal and external system. The gut possesses the largest reservoir of microbes in the entire body. These microorganisms act as a barrier to other bacteria effectively keeping them from traveling outside of the intestinal lumen (Hammer 2015). Learning to appropriately moderate and maintain our microbiome is essential to developing and sustaining a healthy body. Damage that is afflicted to the balance of probiotic microorganisms in the human biome allows pathogenic microbes to easily infiltrate the system and wreak havoc. According to Informa Healthcare, “Widespread overuse of antibiotics has led to the emergence of numerous antibiotic-resistant bacteria; among these are antibiotic-subsisting strains capable of surviving in environments with antibiotics as the sole carbon source” (2016). Now that we have been using antibiotic pharmaceuticals for some time and research has shown the negative impacts that they stand to exhibit we need to address how we might be able to use other compounds in place of pharmaceuticals and whether they will display the same negative effects of long term use of antibiotic pharmaceuticals. Allium sativum and Melaleuca spp. essential oil both have antimicrobial properties, therefore both samples should display zones of inhibition.

Materials and Methods

Using aseptic techniques, a live culture of Kocuria rhizophilia was applied using a sterilized swab to two sterile agar plates using the bacterial lawn method. The agar plates were then divided evenly into three parts using a marker on the backside of the plate. A a paper disk soaked in Melaleuca spp. essential oil was added to the center of one section, an equal sized piece of raw Allium sativum was placed in the second section, and a sterile paper disk soaked in water was applied to the third section as a control. The plates were then labeled and placed in an incubator at 27° Celsius for one week. After the allotted time had passed the cultures were removed from the incubator and the presence of a zone of inhibition was noted and measured.

Results

Both samples displayed very similar results on both tests. On the first plate, Melaleuca spp. essential oil displayed a zone of inhibition of 3cm, whereas Allium sativum displayed a zone of inhibition of 1.5cm, and the control showed no zone of inhibition (see figure 1).

Figure 1

On the second plate, Melaleuca spp. essential oil displayed a zone of inhibition of 3cm, whereas Allium sativum displayed a zone of inhibition of 2cm, and the control showed no zone of inhibition (see figure 2).

Figure 2

Discussion

As we had predicted, both Melaleuca spp. essential oil and Allium sativum effectively displayed antibiotic properties and inhibited the growth of Kocuria rhizophilia while both plates showed the control, water, having no inhibitory effects. While Melaleuca spp. essential oil exhibited the most widespread zone of inhibition, this is somewhat arbitrary in this experiment. The solubility of Melaleuca spp. essential oil and the compounds in Allium sativum were not tested and thus the zones of inhibition are just markers that the samples do exhibit antibiotic properties. As Mateus de Oliveira Negreiros et al. Discovered in their experiment, Baccharis psiadioides essential oil inhibited growth and displayed antibiotic properties against antibiotic-resistant Enterococcus faecalis strains (2015). Most pharmaceutical antibiotic medicines are derived naturally and further engineered in a laboratory. Now that research shows that these engineered medicines are causing detriment and creating strains that even feed off of the antibiotics we are using perhaps we need to refocus our attention towards the roots of these pharmaceuticals and test their efficacy against common pathogens.

Literature Cited

Fiore, D., Fettic, L.,Wright, S., Ferrara, B. (2017). Antibiotic overprescribing: Still a major concern . The Journal of Family Practice, 66(12), 730-736. 0094-3509.

Hammer, A. (2015). The Effects of Alcohol on Post-Burn Intestinal Barrier, Immune Cells, and Microbiome. Current Reviews ALCOHOL RESEARCH, 37(2), pp.1-014. 2168-3492.

Negreiros, M. D., Pawlowski, Â, Zini, C. A., Soares, G. L., Motta, A. D., & Frazzon, A. P. (2016). Antimicrobial and antibiofilm activity of Baccharis psiadioides essential oil against antibiotic-resistant Enterococcus faecalis strains. Pharmaceutical Biology, 54(12), 3272-3279. doi:10.1080/13880209.2016.1223700

Rashid, M., Weintraub, A., & Nord, C. E. (2012). Effect of new antimicrobial agents on the ecological balance of human microflora. Anaerobe, 18(2), pp.249-253. doi:10.1016/j.anaerobe.2011.11.005

Woappi, Y., Gabani, P., Singh, A., & Singh, O. V. (2014). Antibiotrophs: The complexity of antibiotic-subsisting and antibiotic-resistant microorganisms. Critical Reviews in Microbiology, pp.1-14. doi:10.3109/1040841x.2013.875982

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The efficacy of the antibiotic properties of garlic and tea tree oil against Kocuria rhizophilia [Scientific Research Paper]