Thrifty-Bee Honey vs Germ-X brand sanitizer as Escherichia coli growth Inhibitor

Morgan Ries, Sara Seegers, Hannah Bohnaker

Abstract:
This study explores the effectiveness of Thrifty-Bee Honey vs Germ-X brand sanitizer as an antibacterial
used against E.coli. This is important to study because of how common this bacterial species is and since
honey is found worldwide where antibacterials may not be readily available. Due to the vast differences
in area f kill zones within the petri dishes, this experiment shows supportive evidence that Thrifty-Bee
Honey can be used as an antibacterial and may even be more effective than some Germ-X brand
sanitizers.
Introduction:
The purpose of this study was to provide insight into how a natural and inexpensive alternative to
chemical antibacterials can be utilized. Common bacteria have become increasingly resistant to
antibiotics over the years due to overuse of antibiotics. Though some new regulations are developing for
existing medications, progress has been slow in creating new alternative drugs, and this epidemic of
superbugs is becoming a dangerous public health issue (Spellberg 2008). Honey has long been known
to have antibacterial properties, and this study demonstrates how effective it is compared to a
commercially sold antibacterial product such as hand Germ-X brand sanitizer. Honeys antibacterial
effects are mostly thought to come from the levels of peroxide present, but non-peroxide properties like
acidity also contribute (Bogdanov, 1997, p. 752). The peroxide and polyphenols in the honey both inhibit
bacterial growth and degrade the DNA (Brudzynski, 2012, p. 335). Because of these properties, the
primary hypothesis is that because of the multiple, potent antibacterial components in honey, it will be as
or more effective at preventing and killing bacterial growth. The kill zones for the Thrifty-Bee Honey
were predicted to be the same diameter as the kill zones of the Germ-X brand hand sanitizer, and that the
E. coli growth will be completely inhibited in the area that the discs were touching. The experimental
approach was to grow three plates of E. coli and draw and label three sections on each plate. The first
section was used as a control, and exposed one to antibacterial discs soaked in deionized water. The
second was exposed to discs soaked in hand Germ-X brand sanitizer, and the third was exposed to discs
soaked in Thrifty-Bee Honey. Whichever section showed the largest kill zone was the most effective
Materials and Methods:

The antibacterial agents used in this experiment were Germ-X brand sanitizer and Thrifty Bee brand
honey. Six agar plates were obtained and each was sectioned into three equal parts. This was done with
sharpie drawn on the bottom of the plates. Each section was labeled with either a C for control, H for
Thrifty-Bee Honey or A for antibacterial, or in this case, Germ-X sanitizer. Each plate was dated and
labeled so as to be identifiable. The six agar plates were each coated with 200 micro-milliliters of an
E.coli bacterial solution. This was done by pipetting the bacterial solution onto the plates and then
thoroughly spreading it around the entire surface of the plate with sterile swabs. New pipette tips and
swabs were used for each dish. 6 paper disks were soaked in a 50.0 mL beaker with germ-X and 6 paper
disks were soaked the same way in the Thrifty-Bee Honey, 6 were soaked in distilled water for the control
section. The disks were then placed on the corresponding marked area on the agar with tweezers and
lightly pressed down.
Obtain 6 petri dishes, agar plates, E. Coli , and permission to use incubator from Lab instructor.

Obtain Germ-X brand hand sanitizer and Thrifty-Bee brand honey . Soak paper disks in antibacterial agent and DI water as control.

Section off petri dishes so each has labeled for Honey, labeled for Sanitizer, and labeled as the control.

Coat dish with bacterial infused broth, place paper disks in appropriate sections.

Secure lids to the dishes and incubate overnight at 37 Degrees Celsius.

Reaction will be allowed 4 days to occur and the area of dead zones in which the E.Coli was killed will be calculated and graphe

Image 1: colorized scanning electron

micrograph of E. coli (CDC, 2006)

Image 2: The swabbed petri dishes with soaked

paper disks

Results:

Graph 1: The area of the kill zones in both the honey and sanitizer trials are displayed. The diameter
measurements are in centimeters. The honey was clearly more effective by a factor of on average 10.
Because the data is finding the size of a kill zone caused by either Thrifty-Bee Honey or Germ-X brand
sanitizer, it is logical to measure the area of these zones and graph that as the quantitative data. It is easily
shown in a line graph to show the vast differences between the two variables. The line of best fit was
calculated by plugging the information into the formula to calculate linear regression.

For the Germ-X brand sanitizer, the trend was a gradual decrease in effectiveness, though that could be
due to imperfections in putting equal amounts of Germ-X brand sanitizer on each disk and, more often
than not, the measurements were equal.
For the Thrifty-Bee Honey, we also had a gradual decrease, though the overall calculated area was
ridiculously higher than that of the Germ-X brand sanitizer kill zone. This could be due to mistakes in the
original preparation of the disks. The Thrifty-Bee Honey was much less viscous and slid around as the
experiment was set up, though not so much that it would have made that big of a difference.
Though this experiment was harder to quantify in exact measurements and there were likely mistakes
made, it still shows how well Thrifty-Bee Honey works as an antibacterial. The experiment even shows
that Thrifty-Bee Honey works as a superior antibacterial to hand Germ-X brand sanitizer.

Germ-X brand sanitizer

Thrifty-Bee Honey

Mean y (y): 0.75833333333333

Mean y (y): 11.783333333333

Intercept (a): 0.94333333333333

Intercept (a): 12.873333333333

Slope (b): -0.052857142857143

Slope (b): -0.31142857142857

Regression line equation: y=0.94333333333333-

Regression line equation: y=12.873333333333-

0.052857142857143x

0.31142857142857x

Graph 2: Shows the linear regression of the area of dead zones around the Germ-X sanitizer and the
Thrifty-Bee honey.
Discussion:

The purpose of this study was to mediate the problem of antibiotic resistant drugs by demonstrating
the effectiveness of honey as a possible alternative to common antibiotics. The hypothesis was that
because of the multiple, potent antibacterial components in honey, it will be as or more effective at
preventing and killing bacterial growth, and the predictions were that the kill zone diameters would be
the same for the honey disks and the hand sanitizer disks, and that bacterial growth would be inhibited
completely in the areas that the disks were touching. The hypothesis was confirmed by the results that
clearly indicated that Thrifty-Bee Honey works as an effective antibacterial and appears to be more
effective than the Germ-X brand sanitizer used at killing E.coli. The average area of the kill zone as a
result of the Thrifty-Bee Honey was on average 5 times larger than the kill zone resulting from the
Germ-X brand sanitizer in all 6 plates (Graph 1), much larger than predicted. The control section for
each agar plate showed no kill zone and provides a significant control basis. The Germ-X brand
sanitizer induced kill zones were all more symmetrical and evenly distributed than the Thrifty-Bee
Honey kill zones. This discrepancy could be attributed to the honey paper disks sliding around on the
agar more commonly during initial application. This uneven display of the kill zones suggests that the
honey killed the E. coli anywhere it contacted the bacteria. The paper discs soaked in Thrifty-Bee Honey
and Germ-X brand sanitizer did not hold equal amounts of either substance and thus dispersed unequal
amounts of either Thrifty-Bee Honey or Germ-X brand sanitizer onto the agar, this could have
potentially altered the kill zone area. These findings strongly indicate that honey has potential to be used
as a professional antibiotic. One study done by Patricia Lusby, Alexandra Coombes, and Jenny
Wilkinson tested honey as an antibacterial against 13 strains of bacteria. The honey inhibited growth in
all but one bacterial species. This study showed how all various varieties of honey, including processed
and unprocessed, inhibited the bacteria growth. A follow up study comparing the bacteria-killing effects
of Thrifty-Bee Honey to an antibiotic such as penicillin would be necessary to prove that honey is a
viable alternative to medical antibiotics. The procedure would remain largely similar, but antibiotic
disks soaked in the appropriate dosage of penicillin would be used rather than Germ-X brand hand
sanitizer.

Acknowledgements:
Haley Pillars, for her time and effort in and outside of lab.
Dr.Cao, for teaching.
References:

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action: Polyphenol/H2O2-induced oxidative effect on bacterial cell growth and on DNA degradation.
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http://phil.cdc.gov/phil/details.asp?pid=10068
Lusby, P., Coombes, A., & Wilkinson, J. (2005). Bactericidal Activity of Different Honeys against
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