OreganoCH

Notes Remember "Oregano Oil" means "Oregano Essential Oil" and the final report should say "Oregano Essential Oil". Materials:  Oregano Essential Oil  10 Mueller Hinton Plates  20 wafers hole punched from Qualitative Filter Paper and impregnated with Oregano Essential Oil  Escherichia coli stock  Staphylococcus aureus stock  10 cotton swabs  Bunsen Burner with gas  Tweezers  2 beakers  Alcohol  1-piece Qualitative Filter Paper  Hole punch Method: 1. Label 10 Mueller Hinton Plates and draw dividing line halving each plate 2. Punch 20 wafers from Qualitative Filter Paper 3. Drop 20 wafers into 1 beaker and drop Oregano Oil onto wafers until soaked 4. Light Bunsen Burner 5. Collect Stock of E. coli and S. aureus 6. Lawn 5 plates with Escherichia coli (using new cotton swab each plate) 7. Lawn 5 plates with Staphylococcus aureus (using new cotton swab each plate) 8. Considering the dividing line, place one Oregano Oil impregnated wafer into each half of each inoculated plate. (flame tweezers between each wafer placement) 9. Incubate all 10 plates at 37 C for 24 hours. 10. Observe after incubation period. Abstract Introduction A. Background information 1. Introduction to Kirby-Bauer Technique a. Briefly explain Kirby-Bauer technique b. Mention disk diffusion method to determine sensitivity or resistance of bacteria to specific anti microbials 2. Rationale for Choosing Oregano Oil a. Why use oregano oil as antimicrobial agent

b. Discuss known antibacterial, antifungal, antiviral effects c. Highlight previous studies that support oregano oil as antimicrobial agent 3. Background on E. coli and S. aureus a. Provide general information (characteristics, typical habitats, pathogenicity) b. Why choose these to study? 4. Antibiotic Resistance and Alternative Treatments a. Discuss global issue of antibiotic resistance and why alternative antimicrobial b. Emphasize importance of exploring alternative antimicrobial 5. Previous Research on Oregano Oil a. Summarize relevant studies b. Highlight any findings that support oregano oil as antimicrobial 6. Objectives of Your Study a. Clearly state objectives of the study (assessing the effectiveness of oregano oil against .... using .... technique) B. Thesis statement 1. Antimicrobial Efficacy 2. Target Organisms 3. Methodology 4. Broader Context Methodology A. Research design 1. Participants  Name  Name  Name 2. Materials  Oregano Oil  10 Mueller Hinton Plates  20 wafers hole punched from Qualitative Filter Paper and impregnated with Oregano Essential Oil  Escherichia coli stock  Staphylococcus aureus stock  10 cotton swabs  Bunsen Burner with gas  Tweezers  2 beakers  Alcohol  1-piece Qualitative Filter Paper

B. Statistical analyses Materials For the inoculation, we utilized a collection of materials, both independently obtained and provided in the lab. A 15 mL bottle of oregano essential oil was provided by group member James. Within the lab, we were provided several sterile materials for inoculation, including the following: gloves (one pair for each group member), ten Tryptic Soy Agar plates prepared with Muller Hinton Agar, sterile cotton swabs (one swab for each plate), alcohol, and 20 antibiotic wafers from sterile filter paper. Sterile tools were also provided to us, including two Bunsen burners, two sterile beakers, a sterile hole punch, and a pair of sterile tweezers. For the chosen samples of bacteria, we opted to use one sample of gram-negative bacteria and one sample of gram-positive bacteria. Our gram-negative sample was a preprepared Tryptic Soy Broth (TSB) sample of Escherichia coli (E. coli) ; the sample of gram-positive bacteria was a preprepared TSB sample of Staphylococcus aureus ( S. aureus ). Method The Mueller Hinton agar plates were provided to us, where we separated and labeled the plates accordingly to ensure no cross contamination between samples. The plates were divided in half to display the zone of inhibition. Following aseptic techniques, we first prepared our antibiotic wafers, soaking them in a small amount of oregano essential oil and allowing them to sit in the sterile beaker while we prepared the next materials. Given the nature of our test, and our expected result, we utilized the Kirby Bauer method; the Kirby Bauer method would best demonstrate the effectiveness of the oil against the pathogens we selected. To inoculate the plates properly and effectively, we proceeded with a “lawning technique” to fully diffuse the TSB samples of both E. coli and S. aureus . Aseptic techniques were applied in our inoculation by lighting the Bunsen burners and exposing the plate to heat to prevent contamination of the sample and the agar plates. The sterile swabs were used to collect the sample to transfer from the broth to the plate. New sterile cotton swabs were used for each plate to ensure no contamination was introduced. Following the lawning of the sample, the wafers infused with the oil were placed in consideration with the line drawn on the plate, one wafer on both sides of the line. The tweezers were sterilized with alcohol and flamed to eliminate bacteria following the wafer placement. Following inoculation, the agar plates were incubated at 37 degrees Celsius for 24 hours and stored until observation. Discussion Results Our experiment's results show that oregano essential oil has a notable antimicrobial effect, working effectively against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli ). The two bacterial strains' differing levels of efficacy point to variations in susceptibility or modes of action. More research is needed to determine which characteristics of oregano essential oil contribute to its increased

antimicrobial activity against S. aureus, given its greater efficacy against the pathogen. The outcomes are consistent with earlier research showing oregano essential oil's antimicrobial qualities, especially regarding its effect on Gram-positive bacteria. The oil's bioactive ingredients may interact with the cell structures of microorganisms to cause vital processes to be disrupted and result in bactericidal effects. Gaining knowledge of these mechanisms may help develop fresh strategies for preventing bacterial infections. Furthermore, the observed concentration-dependent response emphasizes how crucial dosage optimization is for real-world applications. Additional investigation into concentration-response relationships may help us better understand the therapeutic range and direct the creation of antimicrobial agents based on oregano essential oil. Limitations of the study This study contains some limitations that should be acknowledged, despite the encouraging results. The first possibility is that the experiment's complex conditions might not accurately reflect those of the real world. Isolating variables in a controlled laboratory setting is crucial, but it can oversimplify interactions that affect oregano essential oil's antimicrobial activity. Moreover, our results are not as applicable to a wider range of microorganisms due to our narrow focus on just two bacterial strains: S. aureus and E. coli. To furnish a more comprehensive comprehension of the antimicrobial potential of oregano essential oil, future research endeavors ought to contemplate broadening the scope of examined strains. It is important to recognize the possible impact of outside variables, such as changes in the composition of oregano essential oil and bacterial resistance mechanisms. Variability in bacterial strains and natural products may impair the reproducibility of results and call for additional research. Implications for future research Our findings open new research directions that will help us better understand the antimicrobial qualities of oregano essential oil. Clarifying the precise bioactive substances in charge of the noted effects against S. aureus and E. coli is one line of inquiry. The development of targeted antimicrobial agents with improved efficacy and fewer side effects can be facilitated by identifying these compounds. More research on the molecular and cellular mechanisms of action may shed light on the ways in which oregano essential oil alters the structures and functions of bacteria. This information is essential for developing more potent therapies and tackling foreseeable problems like bacterial resistance. Our results will be more applicable to real-world situations if we conduct studies in more complex environments, like biofilms or host-microbe interactions, and broaden the scope of antimicrobial assays to include other clinically relevant bacteria. As a result, our study concludes that oregano essential oil has a promising antimicrobial potential. However, further research is required to fully realize this potential and tackle the challenges posed by real-world applications. This work contributes to the ongoing efforts to combat bacterial infections by laying the groundwork for a more nuanced investigation of natural products as antimicrobial agents. Conclusion Our study provided compelling evidence supporting the antimicrobial effect of oregano essential oil against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). The differences in effectiveness observed between the two bacterial strains indicate that they may be affected in different ways or have different levels of sensitivity to the oil. Although our findings support previous research highlighting