Copper Alloy Gym Equipment Harbors Less

Bacteria than Traditional Gym Equipment Surfaces

Nathan Zaroban, Annika Helverson, Francesca Varias
12-5-14

Abstract
Gyms are notorious for being a breeding ground for all kinds of bacteria. With
people moving from one piece of equipment to another, bacteria is spread very
quickly all over. Copper alloy could potentially be the perfect solution to this
problem as it is known to have antimicrobial properties. Gym equipment has just
recently started to be made out of copper alloy in attempts to combat the spread of
bacteria. This experiment compares the number of bacteria found on surfaces of
traditional gym equipment (those made of steel, iron, or rubber grips) to equipment
surfaces plated with copper alloy. Similar types of equipment were swabbed for
bacteria for both types of surfaces. In every instance tested, copper alloy surfaces
had extremely less bacterial counts than similar equipment made of traditional
material.

Introduction
Copper alloy has extreme antimicrobial properties that inhibits the growth of
bacteria and other pathogens on its surface (Weber, Rutala 2012). Grass, Rensing,
and Solioz say that this use of coper as a disinfectant has dated back to around
2600-2200 B.C (2011). However it has not been until just recently that is has been
brought back into mainstream commercial use. It has just recently been registered
by the Environmental Protection Agency as the first solid antimicrobial material

(Grass, Rensing, Solioz 2011). As copper alloys legitimacy as an auto-disinfecting

surface has been established, hospitals have started to replace commonly touched
objects with those made of copper. These include bed rails, doorknobs, and
restroom surfaces. Studies have shown that replacing these surfaces in hospitals
with copper drastically reduces the bacterial counts found on these surfaces
(Schmidt, Attaway III, Fairey, et al 2013). Thus, in turn, it can cut down the risk of
Healthcare-Acquired Infections by over 50% (Salgado, Sepkowitz, John et. al 2013).
Copper alloy works, so it should be implemented in many other places, such as
gyms, to reduce the spread of bacteria elsewhere.
It is no secret that gyms are not exactly the most sanitary places. Hundreds
of people come in and use the same pieces of equipment every day. They move
around from equipment to equipment, carrying the bacteria wherever they go and
leaving it elsewhere for someone else to pick up. There is no telling how many
people have touched the same surface before it gets cleaned. Disinfection wipes are
almost always located in various spots around the gym. The intention is for people
to wipe down each piece of equipment before and after every use. However,
humans cannot be trusted to do this each and every time. This is where copper
alloy comes in to play.
We first learned of the antimicrobial properties of copper alloy from Professor
Hinsa-Leasure at Grinnell College when she described her research on copper in
hospitals in lecture. She then informed us that a company named CuVerro is starting
to manufacture gym equipment plated with copper, and that a local gym would be
one of the first to receive its equipment. Being that all three of us frequently
exercise with gym equipment of traditional, germ-ridden materials, we were
interested to see if the antimicrobial effects of copper would work as well in gym

equipment. To test this, we first swabbed for bacterial counts on traditional gym
equipment at the Charles Benson Bear Center gym at Grinnell College. Then we
swabbed for bacterial counts on gym equipment made with copper alloy at the Paul
W. Ahrens gym in downtown Grinnell, Iowa. We hypothesized that the copper alloy
gym equipment would have significantly less bacterial counts than the equipment
made of traditional materials because of coppers antimicrobial properties.

Materials and Methods

Sampling and Plating:
A phosphate buffered saline-lecithin-Tween 80 (PBSLT) solution was made by
mixing 8 g of NaCl, 0.2 g KCl, 1.44 g Na2HPO4, 0.24 g KH2PO4, 0.7 g Lecithin, and 5.0
ml Tween 80 per liter. The mixture was autoclaved for 20 minutes until the pH was
between 7.2 and 7.4. This solution was used for multiple purposes throughout the
experiment.
15 6x1 inch swabbing wipes were cut and each put into a 50ml conical tube.
The tubes were labeled by equipment type and sample number (Ex. 15 dumbbell 1,
15 dumbbell 2). 600 l of sterile PBSLT was added to each wipe in the tube. Using a
sterile glove, a sterile wipe was removed from the labeled conical tube and wiped
on the corresponding piece of equipment. A specific area of the piece of equipment
was predetermined for the swabbing. The area was thoroughly swabbed five times
with back and forth motions. This sampling procedure was carried out for each of
the 15 pieces of equipment. Four sets of samples from each piece of equipment
were taken from the copper gym. Three sets of samples of the traditional equipment
from the Charles Benson Bear Center gym at Grinnell College, except for the

dumbbells, for which four samples were taken. The sampling occurred over the span
of four weeks.
In the lab, 6 mL of sterile PBSLT were added to each conical tube and
vortexed at the highest setting for one minute. Dilutions sets of 10^0 and 10^1
were prepared from the vortexed sample solution. The 10^0 dilution was made by
transferring 1 mL of a labeled sample solution into a labeled Eppendorf tube. For
the 10^1 dilution 100 l of labeled sample solution was added to 900 l of sterile
PBSLT in a labeled Eppendorf tube. This dilution process was carried out for each
sample.
The 10^0 and 10^1 dilutions for each sample were plated on TSA/5% sheep
blood agar plates and incubated for 48 h at 37C. The plates were counted at 24
hours in case there was rapid growth preventing countable colonies at 48 hours.
Data was collected at both 24 hours and 48 hours, but only the 48 hour data was
used.

Polymerase Chain Reaction amplification of 16S rRNA gene and sequencing

(PCR):
Further research was done to determine the strains most commonly found on
both surfaces. Isolates were streaked onto TSA/5% sheep blood agar plates and
incubated for 48hrs. An isolated colony was taken and mixed into a PCR tube
containing 20l of GeneReleaser. This was done for each of the different isolates
that were streak plated. The PCR tubes were run in a GeneReleaser program to
bring the template DNA at the top of the PCR tube in an aqueous layer. The
reagents were added for PCR to the top layer. The PCR reagents (10 l nuclease
free water, 1.25 l forward primer, 1.25 l reverse primer, 12.5 l master mix) were

added to the top layer. The PCR tubes were put into one of the thermocyclers set for
the 16S program.
A 1% agarose gel was made in TBE. The gel was made by adding 0.35 g of
agarose to 35 ml of TBE and then microwaving the mixture of 20-30 seconds. 3 l of
SYBR safe was added and the gel mixture was poured into a gel box and a comb
was added in the top of the gel.
10 l of the aqueous layer in the PCR tube was removed and mixed with 2 l
of loading dye. The mixture was run on the 1% agarose gel. This was done for each
PCR tube. Then 9 l of Kb ladder was run in an empty lane. The gel was run for 30
minutes at 100 volts. A picture was taken of the gel using the Geldoc system. The
genus of the organism was then determined with the Ribosomal Database Project
web-based software.

Results
This experiment compares the number of colony forming units per 100
square centimeters found on standard gym equipment to similar equipment made
of copper alloy. For instance, data for 15 pound dumbbells made of traditional
material is compared to data for 15 dumbbells made of copper alloy. A t-test was
completed for each similar type of equipment tested to ensure that the data was
significantly different. For each comparison, the copper alloy surface was found to
have significantly less bacteria on it than the traditional surface. The average
decrease in bacteria on copper material from traditional was approximately 94.96%.
From the data we see that the least difference between the two occurred in the

kettle bell weights. We believe this is due to the fact that the traditional kettle bells
were fairly new to the Bear gym and thus less people used them frequently.

For the PCR analysis, five different strains were tested. Both the first and fifth
strains were found to most closely match Micrococcus yunnanensis or Micrococcus
luteus and were common on both surfaces. The second strain matched with Bacillus
altitudinis and the third strain matched with Bacillus pumilus. Both of these strains
were common on both types of surfaces. The fourth strain, Staphylococcus sp.
H780, was found frequently in the Bear gym. Other cell lysing bacteria was present,
but this was the most common. Various kinds of lysing bacteria were also found on
the copper surfaces, however. Just from observations, it seemed that the copper
alloy surface did not effectively kill Staphylococcus bacteria as well as it did other

types of bacteria. We did not test this as our original question only addressed total
bacterial counts. The Staphylococcus found on copper alloy equipment varied in
size, shape and color, so we did not select one to analyze.

Discussion
As hypothesized at the beginning of our experiment, there was a drastic
difference between the bacterial counts on the traditional equipment surfaces and
the equipment made of copper. In most cases copper was found to have 98-96%
less bacteria than its traditional equipment counterpart. This is close to the 99.9%
reduction found by clinical trials run by the EPA (Michels and Anderson, 2008).
It must be noted that there is a difference in the amount of use that each
gym gets. The Bear gym receives around 400-500 visitors a day, while the Paul W.
Ahrens gym has about 100-200 visitors every day. That difference in traffic most
likely does play a role in the difference in the bacterial counts. Less bacteria is
introduced into the copper equipment facility than the traditional equipment facility.
However, the extreme difference in bacterial counts points to the fact that the
antimicrobial properties of the copper is by far the largest factor.
Copper Alloys antimicrobial properties are obviously powerful and will most
likely work for any use. However, one of our most intriguing observations during this
experiment was that the copper alloy seemed to have less effect on the growth of
cell-lysing bacteria. There seemed to be a slight decrease in cell lysing bacteria
from traditional equipment to copper equipment, but it was not near proportional to
the difference in total bacterial counts. This would be an issue because cell-lysing
properties are typical of Staphylococcus bacteria, which have the potential to be
extremely harmful. For instance, Methicillin-Resistant Staphylococcus aureus (MRSA)

is a potentially deadly staphylococcus bacterial infection (Huang and Platt, 2003).

We did not record data for this because our experimental question only dealt with
total bacterial counts. This should be the next direction for this experiment. If
copper alloy does not kill cell-lysing Staphylococcus bacteria, then perhaps it is not
as magical is we would like it to be.

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