A New Possible Alternative Solution to the “Rising Plague” of Antibiotic Resistance

Lauren Proano
Lee et al. “The Antimicrobial Activity of Essential Oil from Dracocephalum foetidum against Pathogenic Microorganisms.”
Upadhyay et al. “Screening of Antibacterial Activity of Six Plant Essential Oils Against Pathogenic Bacterial Strains. ”

INTRODUCTION METHODS RESULTS

Antibiotic resistance has become a growing problem since the introduction of antibiotic drugs in the 1940s. Antibacterial activity was measured by MIC (minimum inhibitory concentration), In Upadhyay et al.’s study, the MIC and MBC levels for 6 essential
The first evidence of bacterial resistance was seen within a year of the introduction of the first antibiotics, and MBC (minimum bactericidal concentration), and growth inhibition zone diameters. oils were compared to 3 synthetic antibiotics. Lower MIC and MBC
today we are quickly running out of drugs to treat resistant infections that are caused by microbes such as MIC measures bacteriostatic activity (inhibited bacterial growth but not death) and levels reflect higher susceptibility against the bacteria. For each of
MRSA. The future of antibiotic treatment must not only include an increase in new drug development, but MBC measures bactericidal activity (bacterial death). the bacterial strains evaluated, at least 1 essential oil scored equal
also further research in antibiotic alternatives. The Rising Plague by Brad Spellberg presented a very honest In order to accurately test the antibacterial effects of the oil against present to or lower MIC and MBC levels than the 3 antibiotics (Figure 1).
and in-depth look at the global concern of antibiotic resistance and what needs to be done to reverse this issue, antibiotics, it was necessary to find the lowest amount of oil that would produce Bavchi oil was more bactericidal to S. aureus and L. acidophilus
with a special focus on MRSA and its danger to the human population. This lead me to research recent studies inhibitory and bactericidal effects. This was done by diluting essential oils in broth than all antibiotics tested. Similarily, S. pneumoniae had lower
on effective antibacterial alternatives. media and screening them against each bacteria in increasing concentrations. Both MIC levels with clove and ajwain oil than all three antibiotics, and
studies used Filter paper disc agar diffusion assay to determine MIC and inhibition lower MBC levels with ajwain and neem oils. Further, several
zone diameters. inhibition zone diameters were significantly larger from essential
Filter paper disc agar diffusion is a simple method in which the bacterium is spread oils than from antibiotic drugs.
ESSENTIAL OILS as on each agar plate and essential oil is added to small filter paper discs that are placed Both Gram + and Gram – bacteria were susceptible, due to the
ANTIBACTERIAL AGENTS in the middle of the agar plate. After incubation for 24 hours each, the zones of volatility of the oil’s components.
growth inhibition for each bacterium are measured. Sterile water was used as a As little as 2 ul of 100% D. foetidum essential oil completely
Lee et al. and Upadhyay et al. were two separate studies investigating the Essential Oil Structure and Function negative control, and broad-spectrum antibiotics (ampicillin, tetracycline, inhibited all Gram + bacterial growth. Less effective on Gram –
antimicrobial activity of specific naturally occurring essential oils from various plants. ciprofloxacin, and hygromycin B) were used as positive controls. bacteria, which is more resistant due to presence of outer
Common pathogenic bacteria, including MRSA, were used to test for antimicrobial Dracocephalum foetidum In Lee et. al, volatility was also measured with a paper disk saturated in D. foetidum lipopolysaccharide layer.
effects.
39.19% N-Mentha-1,8-dien-10-al and 17% and placed underneath the lid of the petri dish containing the bacteria being tested Most notable result of D. foetidum antibacterial activity was that it
Essential oils are complex mixtures of volatile secondary metabolites that mainly
consist of carbohydrates, alcohols, ethers, aldehydes, and ketones.
Limonene (oil was approx. 4 mm from bacteria). Inhibition zones were measured in the same showed equal effects on the 2 MRSA strains as compared to
The majority of the oils are nonpolar, and due to the hydrophilic nature of the manner. normal S. aureus strains.
Nonpolar, hydrophobic components permeate To measure MIC and MBC further, essential oil dilutions were added to fresh bacteria
bacterial cell wall, the hydrophobicity of the oil enables it to partition in the lipids of
the bacterial cell membranes and mitochondria, thus disturbing the structure and
bacterial membranes and it also inhibits microbial cultures (1:150, bacteria to media) in triplicate and plated for growth. The lowest
making the bacteria permeable. respiration and increases ion leakage. concentration at which no visible growth was obtained in agar plates was considered
Essential oils may also successfully inhibit microbial respiration as well, which leads The main components of this oil have much the MBC. MIC was determined by adding p-INT dye (stains bacteria) to the plates; the
to increased ion leakage. Gram + bacteria were extremely susceptible, due to the simpler structures and mechanisms of inhibition lowest concentration at which no red color appeared was considered the MIC.
absence of a lipo- polysaccharide layer that functions as a barrier in Gram – bacteria. than most synthetic antibiotics, but prove to be
Lee et. al explored the antimicrobial activity of the Essential Oil from Dracocephalum equally, if not more, effective. The nonpolar CONCLUSIONS
foetidum, a herb in Northeast Asia that has long been used in Mongolian traditional structures are also highly volatile, and are therefore
medicine. The antimicrobial activity was measured for 5 gram+ bacteria strains, one able to be released into the air in significant Future implications for these essential oils should include
gram- bacteria strain, and two yeast fungi. The bacteria of interest were B. subtilis, S. amounts. This characteristic is likely due to the N-
aureus (one normal strain, and two MRSA strains), M. lutens, E.hirae, S. mutans, and
therapeutic agents; they have a high potential for
Mentha-1,8-dien-10-al, and greatly increases the herbal/alternative medicine. There is no FDA approval for the
E.coli. effectiveness of antimicrobial activity. In the study,
Upadhyay et. al investigated six plant essential oils, including citrus, olive, ajwain, therapeutic use, and testing has not yet been done to
volatility was specifically measured and compared
almond, Bavchi, and neem. The bacteria tested were L.acidophilus, Streptococcus determine the antibacterial effects of essential oils in vivo;
pneomoniae, S. aureus, M. luteus,B. cereus, K. pneumoniae, and E. coli.
to the diameter of the inhibition zone. Even when
the essential oil did not come in direct contact with
several factors would need to be considered before these oils
the bacteria, it was still equally potent. could be used on humans. The side effects of these oils in
humans are yet to be determined, as possible toxic effects of
the oil’s components could be dangerous and possibly
deleterious for our normal, healthy cells. Also, the minimum
Antibiotics Structure and Function concentration of oil necessary to be effective and not
Both studies used broad spectrum antibiotics as positive controls for the antibacterial effects of the essential oils. Ampicillin and dangerous would have to be determined. Another possibility
Hygromycin B were positive controls for D. foetidum. Tetracyline, Ampicillin, and Ciprofloxacin were positive controls for the six
for further research could be synthetic production of the
essential oils used in the Upadhyay et. al study. These antibiotics were chosen because of their diverse abilities to inhibit bacteria.
Ampicillin is a competitive inhibitor of transpeptidase, a bacterial enzyme that cross-links the peptidoglycan chains to form rigid
necessary components of these essential oils.
cell walls. Causes bacterial cell lysis in Gram+ and some Gram– cells.
Tetracycline inhibits cell growth by inhibiting bacteria translation; it is a protein synthesis inhibitor, inhibiting the binding of tRNA Due to the high volatility of the essential oil components,
to the mRNA-ribosome complex. Cells become resistant to tetracycline by enzymatic inactivation, efflux, and ribosomal protection. especially in D. foetidum, they may be especially beneficial as
Ciprofloxacin kills bacteria by interfering with the enzymes that cause DNA to rewind after being copied, which stops DNA and antiseptic products, such as cleansing formulas and
protein synthesis. disinfectants. This would eliminate much of the concern for
Ciprofloxacin is active against both Gram-positive and Gram-negative bacteria. Numerous pathogens, including S. aureus, Figure 1. Comparison of the most effective essential oil and most effective antibiotic toxic side effects and could be implemented into use sooner.
enterococci, and S. pyogenes now exhibit resistance. against seven bacterial strains, measured by Minimum Inhibitory Concentration Levels
Hygromycin B is an aminoglycoside that kills bacteria, fungi and higher eukaryotic cells by strongly inhibiting protein synthesis (ul/mL). Lower MIC values reflect antibacterial effectiveness; essential oils had Further research has been done with other oils, but often the
through a dual effect on mRNA translation. significantly lower MIC values compared to broad spectrum antibiotics for each same bacterial species are tested against; it would be beneficial
bacteria tested, except K. pneumoniae. Stars represent more than one essential oil or to test many other bacterial species for antibacterial effects.
antibiotic that had this low MIC value.

REFERENCES
Lee et al. “The Antimicrobial Activity of Essential Oil from
Drracocephalum foetidum against Pathogenic Microorganisms.” Journal
Ampicillin of Microbiology, p. 53-57 (2007).
Tetracycline
Ciprofloxacin
Upadhyay et al. “Screening of Antibacterial Activity of Six Plant Essential
Hygromycin B Limonene Oils Against Pathogenic Bacterial Strains.” Asian Journal of Medical
N-Mentha-1,8-dien-10-al Sciences 2(3): 152-158 (2008).