Hauer Serraiocco 1

Introduction
Human hands are used every single day; hands do a wide range of actions from
the simplest of tasks like picking up a piece of paper to the hardest tasks of assembling a
computer. Without hands life would be very difficult. However, hands are also a very
good transportation system for bacteria. In fact, 80% of infectious diseases are spread
through touch. This can easily be solved by washing hands, but through a survey
conducted by the CDC, 67% of the people who said they wash their hands actually do not
(Hardy).
Washing hands is a very vital part in an every day routine. This important step can
protect from harmful diseases and also improve overall health. With so many people in
constant contact with each other it is very important to keep bacteria low and to make
sure that harmful diseases are controlled. Mild to warm water is much more effective at
removing visible oils and dirt than cold water (Barrymore). Using a warmer temperature
of water can kill many more bacteria colonies.
Bacteria likes to grow in warmer places because the heat causes the enzymes in
the bacteria to speed up, thus causing the reproduction process to go much faster. On the
other hand, for some bacteria, if the temperature is too high then it will burn and kill the
cells (Santa). Most people keep their hands in their pockets or tucked under their arms
when waiting around. This can be a potential breeding ground for bacteria. With so many
bacteria there must be a solution. This solution is to wash hands and wash them
effectively. This experiment is to test best possible way to wash hands using a
comfortable temperature of 37 Celsius and the duration of ten seconds as the minimum
and the warmer temperature of 43 Celsius and longer time of 30 seconds as the

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maximum. These two proved to be just right for washing hands comfortably. Washing
hands under these conditions and then testing for bacteria is the best way to collect data.
Knowing that warmer temperatures kill bacteria it is logical to assume that the higher
temperature and time is the best choice to wash any hands. This can also be used to
assume the colder water will allow more bacteria colonies to survive.
Time is one of lifes many factors in the sense that everybody wants more of it.
It is no different here. If the subject is allowed more time to do something then usually
there is a more favorable outcome. For hand washing, it is advised to wash longer in
order to clean more bacteria off (Bass, Finke). This applies to the experiment because the
subject who is performing it has a longer time to get to those hard to reach places such as
the fingernails and in between each finger, this means that when the hands are swabbed
for data it is likely that less bacteria will be found.
As stated before, a majority of bacteria like to grow in warmer places because of
how it speeds up the process of reproduction. The temperature that the majority of
bacteria grow in was found out to be 40C (Smith). This is so because most bacteria are
able to carry out life functions around that temperature. Overall, temperature should have
a positive effect on bacteria growth because only a slight percent will die from the heat
conditions when most will find it more than suitable for reproduction.
After a test subject is swabbed and 24 hours have gone by with the bacteria in its
growing conditions, colonies will then be counted. The data that is being recorded is the
number of colonies that each hand washing combination produces. This will allow the
researchers to figure out the best possible way to wash hands and produce a cleaner
population.

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When a test subject goes through the prepared hand washing procedure, one pump
of hand soap will be applied and then the subject will wash in the desired temperature for
the desired duration. After that, the cleansed hands will be swabbed and put in a test tube
of one ml of distilled water. Finally, the water will be dumped into a Petri dish filled with
agar and 24 hours will be waited, then the bacteria colonies are counted.
Soap helps to kill bacteria by breaking down the oils and fats that are present on
hands. The soap can effectively remove the oils and fats allowing the water to kill
bacteria (Fuller). Bacteria can be harmful and can cause serious illnesses. The most
common bacteria on a human hand are Staphylococcus aureus (Eppes) or staph for short.
This bacteria can cause a staph infection by entering the blood stream through a cut or
wound on the hands. Just washing with water is not as effective as washing with soap.
Water can not break down the oils and fats that hands contain and therefore cannot kill
bacteria (Fuller).
Time and temperature effect the test subject beyond science because most people
in the world either cannot handle hotter temperatures or choose to use a lower one, or just
do not have the patience to perform a full hand wash. With the information stated above,
the researchers believe that when time and temperature are combined it will have a great
effect on hand washing.
Problem Statement
Problem:
Does time and temperature affect how clean your hands are after a hand washing
session?

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Hypothesis:
If hands are washed in 43C for thirty seconds, it will produce the cleanest pair of
hands.
Experimental Design
Materials
Distilled Water
(60) Petri Dish
Stopwatch
Meijer Brand Hand Soap
(60) Sterile Cotton Swabs
Carolina brand Agar (11.5 g)
1000 ml Flask
Test Tube Holder

1 ml Sterile Dropper
Hand Towel
(12) Sterile Test Tubes
(1) 200 ml Beaker
Thermometer
Beaker Holder
Ruler
Teflon Magnet
Hot Plate

Procedure:
General:
1.

Turn on the water.

2.

Let water run for 15 seconds and then fill a beaker up with the running tap water.

3.

To create the agar (see fig. 1) use 11.5 grams of the agar powder.

4.

Fill the 1000 ml flask with 500 ml of tap water.

5.

Add the magnet to the flask of water.

6.

Add the 11.5 grams of agar powder to the flask and place on hot plate. Turn the

hot plate to 6 and wait until the agar is a golden color.

8.

Once the agar is ready fill all Petri dishes needed.

9.

Use the sterile dropper to take 1 ml of distilled water.

10.

Drop the distilled water into the test tube and cover the tube with tin foil then
place the tube in the test tube holder.

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(+,+)
1.

Take a thermometer and measure the water in the beaker to make sure it is 43 C.

2.

Next, wet hands in 43 C temperature and apply one pump of soap.

3.

Rub hands together, cover hands completely in soap.

4.

Wash hands for 30 seconds.

5.

Rinse hand thoroughly in same water temperature.

6.

Perform analysis steps below.

7.

Dry hands with hand towel.

(-,+)
1.

Take a thermometer and measure the water in the beaker to make sure it is 37 C

2.

Next, wet hands in 37 C temperature and apply soap.

3.

Rub hands together, cover hands completely in soap.

4.

Wash hands for 10 seconds.

5.

Rinse hand thoroughly in same water temperature.

6.

Perform analysis steps below.

7.

Dry hands with hand towel.

(+,-)
1.

Take a thermometer and measure the water in the beaker to make sure it is 43 C.

2.

Next, wet hands in 43 C temperature and apply soap.

3.

Rub hands together, cover hands completely in soap.

4.

Wash hands for 10 seconds.

5.

Rinse hand thoroughly in same water temperature.

6.

Perform analysis steps below

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7.

Dry hands with hand towel.

(-,-)
1.

Take a thermometer and measure the water in the beaker to make sure it is 37 C

2.

Next, wet hands in 37 C temperature and apply soap.

3.

Rub hands together, cover hands completely in soap.

4.

Wash hands for 10 seconds.

5.

Rinse hand thoroughly in same water temperature.

6.

Perform analysis steps below

7.

Dry hands with hand towel.

(Standard)
1.

Take a thermometer and measure the water in the beaker to make sure it is room
temperature.

2.

Next, wet hands in 40 C and apply soap.

3.

Rub hands together, cover hands completely in soap.

4.

Wash hands for 20 seconds.

5.

Rinse hand thoroughly in same water temperature.

6.

Perform analysis steps below

7.

Dry hands with hand towel.

Analysis:
1.

Take a distilled swab and swipe the palm, between fingers, and on the back of the
hand

2.

Put swab in a tube of 1 ml of distilled water and twist it around for 10 seconds.

Hauer Serraiocco 7

3.

Pour water onto a Petri dish (see fig. 2 below) and spread it around with the

swab.
4.

Record the number of bacteria colonies after setting the Petri dish in a 37 C
incubator (see fig. 3 below) for 24 hours.

5.

After waiting for 24 hours record the bacteria colonies by counting each
individual dot of bacteria.

6.

If the bacteria is a lawn then find the surface area of the Petri dish using the

formula pi*r2 where r is the radius.

7.

Estimate the percent of area the lawn occupies and multiply it by the surface area.

8.

Find the average size of colonies by measuring the length of five colonies in cm
then adding the five lengths together and then dividing by five.

9.

Divide the number received in step 7 by the average size of a colony found in step
eight and that is an estimate of how many colonies are in that lawn

Diagram:

Figure 1. Carolina Brand Agar

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Figure 2. Bulk of the materials used

Figure 3. 37 incubator where Dishes were Stored for 24 hours

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Data and Observation

Data:
Table 1
Bacteria Colonies
Bacteria (Colonies)
DOE
(Time, Temperature)
(+,+)
(-,-)
(+,-) (-,+)
Standard
1
75
70
82 328
328
2
18
2
295 328
7
3
389
42
129 307
347
4
442
328
279 253
73
5
235
141
74 531
628
6
432
439
405 207
202
7
36
983
352 158
354
8
456
418
432 176
137
Average
260.375 302.875
256 286
260
As shown in Table 1, eight DOEs were ran in the order of (+, +), (+,-), (-, +), (-,-),
and then ended with the standard. The DOE results were not randomized which may have
lead to some bias results in our DOE.

Table 2
Design of Experiment Values
Time ( Seconds )
-

Temperature ( C )

Standard
10

+
20

30

Standard
37

+
40

As shown in Table 2 the levels for each variable were determined through
research. Many hand washing experiments that were conducted by various hospitals and
scientist were researched. These levels were found to work the best with human hands.

43

Hauer Serraiocco 10

To prepare, measure the temperature of the water and use a stopwatch to record the time
of washing hands (Santa).
Observations:
Table 3
Observations of DOEs ran in class
Date
Observation
The Agar was made, the color was a golden brown, and it
3/19/12
had an acidic smell. The Petri dishes would slide around on
the table when bumped. The first DOE was then run.
The first DOE was analyzed. The Petri dishes were
3/20/12
relatively cold. There were many bacteria colonies on each
Petri dish. The second DOE was run
The second DOE was analyzed. It smelled like old gym
3/21/12
socks. The water temperature for the experiment was just
hot enough to sting. The third DOE was then run.
The first lawn appeared in the third DOE. The lawn was a
3/22/12
snot color and filled most of the dish. There were also
strands of multiple bacteria colonies linked together.
DOE three was analyzed and recorded. It smelled like
rotten fish. The DOEs would go into the incubator stacked
3/23/12
on top of each other. Most of the dishes were a faded
yellow color.
DOE four was run on this day. When multiple sinks were
running at the same time, the temperature would me
3/26/12
thrown off. Each subject used only one pump of soap. No
other DOE were analyzed.
DOE five and six were run. Many lawns showed up in
3/27/12
DOE four. This may be due to having bio class last on that
day.
DOE five and six were analyzed and recorded. There were
a small amount of bacteria colonies in the (-, +) dish of
3/28/12
DOE five. Other dish also had the linked colonies. DOE
seven and eight were run.
On the final day DOE seven and eight were analyzed. The
(+, +) dish of DOE seven had even few bacteria colonies
3/29/12
the DOE five's (-, +) dish. The color of the dishes was more
clear then yellow.

As shown in Table 3, the dates of 3/28/12 and 3/29/12 show that there were low
results in DOE five and seven with the variables (-,+) and (+,+) respectively. As shown in

Hauer Serraiocco 11

Table 1, the fifth DOE trial result for the variable (-, +) is 74, which is considerably low.
Also shown in Table 1 is the seventh DOE trial result for the variable (+, +) is 36, which
was even lower than the DOE fives (-, +). As shown in Table 2, the first variable was a
time of ten seconds and the + variable was 30 seconds. The second + variable was a
temperature of 43 Celsius.

Trial
Standard

Trial
(+,-)

Trial
(-,-)

Trial
(+, +)

Trial
(-, +)
Figure 4. One Full Set of Petri Dishes for One DOE

Figure 4 above shows the first DOE that was run on March 19th. The way
that bacteria was counted was that every time a new colony was found it was dotted with
a black marker so it would have been easy to identify which ones had been counted and

Hauer Serraiocco 12

which ones had not. The (+,-) combination had the least amount of bacteria for this
specific trial. The (-,-) combination had the most bacteria because it was a lawn.
A lawn is when bacteria cover a large portion of the dish. This means that
the counting methods had to be adjusted. The dishes used were measured to have a
diameter of 10 cm and the surface area for a circle is A= r2 where r is the radius. Pi was
rounded to 3.14 as learned in math class. Since the diameter is 10 the radius is 5 because
the radius is half the diameter. When the formula is solved the surface area for the Petri
dish was 78.5 cm2. It was decided that the bacteria colonies would be measured in
millimeters so the 78.5 was converted to 785 by multiplying it by 10 because the ratio of
a centimeter to millimeter is 1:10. The average size of one colony of bacteria was 2.4
mm. If a dish was covered completely by a lawn the total number of bacteria colonies
would be 328 because if the total surface area is divided by the average colony size the
quotient is the number of colonies in that lawn. If a lawn did not cover an entire dish,
then an estimated percent would be determined for that specific lawn. Once that percent
is decided it is multiplied by the total surface area of 785 mm2 and then it is divided by
the average colony size to get the number of colonies in that lawn.

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Trial (+, +)

Figure 5. Representative Petri Dish From (+, +) Trials

Figure 5 shows the Petri dish results for the (+, +) DOE. The black dots on the
dish itself were marks from a Sharpie. These marks helped aided in the counting of each
bacteria colony. This dish was a (+, +) dish which means the high amount of time of 30
seconds and the high amount of temperature of 42 Celsius.

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Trial (+,-)

Figure 6. Sample Petri Dish From (+,-) Trials

As shown in Figure 6, there is a bacteria strand near the word trials on the Petri
dish. This strand was multiple bacteria colonies linked together. When measuring these
colonies simply look closely at the strand and notice that there are individual circles of
colonies. Count each circle as its own colony.

Data Analysis and Interpretations

Data Analysis:
Table 4
Standards of Bacteria Colonies
328

347

Bacteria Colonies
73
628

202

354

137

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Figure 7. Range of Standards

In Figure 7 the table shows the standards, which for the most part, are within a
close range of each other. The only exception is the day where only 7 colonies were
recorded. This could be due to the fact that some people might have not been in contact
with many things that day, or the bacteria just could not grow in the environment it was
in which is odd because it is the only single digit result that was recorded. This could also
imply that the experimental design was flawed.

Table 5
Effect of Time on Bacteria Colonies
Time
(-)
(+)
302.875
260.375
286
256
Avg = 294.4375 Avg =
258.1875

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Figure 8. Effect of Time on Bacteria Colonies

As shown in Figure 8 and Table 5 the less time spent washing hands the greater
the amount of bacteria colonies that will grow. The effect of Time is -36.25. On average,
as time increases, the growth of bacteria colonies decreases by 36.25 colonies.
Table 6
Effect of Temperature on Bacteria Colonies
Temperature
(-)
(+)
302.875
260.375
256
286
Avg = 279.4375 Avg =
273.1875

Figure 9. Effect of Temperature on Bacteria Colonies

Hauer Serraiocco 17

As shown in Figure 9 and Table 6, the lower the temperature used the higher the
amount of bacteria colonies that will grow. Effect of Temperature is -6.25. (The change
from low to high) On average, as temperature increases, the growth of bacteria colonies
decreases by 6.25.

Figure 10. Effect of Time and Temperature on Bacteria Colonies.

Table 7
Effect of Time and Temperature on Bacteria Colonies

Time

Temp.
(-)

(+)

Solid
Segment

(+)

256

260.375

Dotted
Segment

(-)

302.875

286

As shown in Figure 10 and Table 7 the line segments do not cross which mean
there is no interaction point.
Slope of the segment of time (+) minus Slope of the segment of temperature (-)
gives the Effect (time vs. temperature) = 4.375/2-(-16.875/2) = 10.625. This means that
the interaction effect between time and temperature is 10.625 bacteria colonies.

Hauer Serraiocco 18

Interpretation:
Figure 8 is showing the effect of time on bacteria colonies. The slope is clearly going
downward which means that the more time spent washing your hands the less the amount of
bacteria colonies that will grow. The number 294.4375 represents the amount of average bacteria
colonies grown when using the low amount of time washing and the number 258.1875 represents
the average amount of bacteria grown when using the high amount of time washing. When the
procedure was ran with 30 seconds it had a lower average bacteria count and this could be due to
the fact that since more time was allowed, more bacteria could be cleaned from the subjects
hand compared to a shorter amount of time.
Figure 9 is showing the effect of temperature on bacteria colonies. The slope in this graph
is also going downward. This means that when washing your hands in higher temperature water
less bacteria colonies will grow. This was result was rather odd because after doing research, it
was discovered that bacteria actually grows best in warmer atmospheres because as heat
increases, enzyme activity speeds up and causes the bacteria to divide faster, but it was also
discovered that some bacteria grow best in colder atmospheres because if the heat is too high it
will kill the bacteria (Santa). The number 279.4375 represents the average amount of bacteria
colonies grown when using the lower temperature and the number 273.1875 represents the
average amount of bacteria grown when using the higher temperature.
The graph in Figure 10 shows no interaction. This means that the two variables of time
and temperature are not related in this experiment and there is no combined effect on the
response variable of bacteria colonies. As shown in Table 7, the combination of (+,-) and the
combination of (+, +) (solid segment) has a positive slope, while the combination of (-,-) and the
combination of (-, +) has a negative slope.

Hauer Serraiocco 19

Figure 7 shows the standards of the experiment. The importance of a standard group is to
have something to compare the high and low results to. However, a standard group is not a
control because the procedure is still being run, in a control; no procedure would be run on that
group. The data recorded is varied greatly because the subject tested was different every time and
due to the activities that subject performed, more or less bacteria could be found on those hands
than on another subjects hands. As a result of this, no set pattern could be found in the data.
Neither time nor temperature were considered vital in this experiment. To determine this, the
range of standards must be found. The highest standard of 628 was subtracted by the lowest
standard of 7 bacteria colonies to get 621 bacteria colonies. Then that number must be multiplied
by 2 to get 1242 to determine if the variable has to be greater than or equal to 1242. In this case,
neither effect values were greater than or equal to 1242.

Conclusion
The original hypothesis was rejected. The researchers believe that the higher temperature
and longer time would be the most effective way to kill bacteria. Bacteria cannot survive in high
temperatures so it is logical that the longer the time spent in higher temperature water the more
bacteria that would die. This did not happen within the experiment. The testing proved that lower
time and higher temperature was the most effective way to kill the bacteria colonies. This may be
this way because when washing hands in high temperature more bacteria will die and the lower
amount of time spent in the water may cause more bacteria to die also.
This experiment was to find out what range of water and time were best for
washing hands. The plan was to wash a total of sixty hands and record the results. This was run
because of the interest in the efficiency of hand washing. The results of this experiment was a

Hauer Serraiocco 20

very high amount of bacteria colonies grown in the low, low variables and a low amount of
bacteria colonies grown in the high, high variables. These results were achieved because of the
varying temperatures of the water.
A design flaw that had occurred was that the DOE itself was not randomized. This
may have created bias results. This could have been fixed with the aid of randomizing software.
Originally, sixty hands were to be washed and tested but due to the lack of time only forty hands
were tested. With better preparations and time management, sixty hands could have been tested.
While running the procedure, some problems were encountered. These problems include
test subjects taking more than one pump of hand soap, drying hands before being swabbed, and
subjects canceling the hand wash because of not being able to withstand the temperature. These
errors were all encountered within the first two days of testing. The reason each of these interfere
with the data collection is because if a person has more soap than others, it will affect how much
bacteria colonies grow. The way to prevent this was to watch each subject carefully to ensure
that the correct amount of soap was applied each time. Next, when a person used the hand towel
before being swabbed it caused the bacteria from the hand towel to move to the persons hand,
thus causing more bacteria to be on the persons hand. This was prevented by one researcher
watching the towel in the supply box thoroughly to make sure to one touched it and only let the
subject use it after the procedure was compete. Finally, when people aborted the procedure
halfway through because of the hotter temperature of 43C it caused the subjects hand to be
somewhat cleansed, therefore that set of hands can no longer be used for data collection. The
way to prevent this is to have the test subject test the water temperature on the forearm to see if
the heat was too intense, and since the forearm is not swabbed it did not affect data.

Hauer Serraiocco 21

Further research could have been done to improve this experiment. Things such as the
cleanliness of the water being used. For example, if the place where the experiment was
conducted had a problem with the water and it gets contaminated it will greatly affect the data
because the bacteria from the water would transfer to the subjects hands (Noorani).Also, if the
subject had rough or soft hands factored into the data because in order to obtain rough hands the
person must be exposed to harsh chemicals (Secor). A person with dry hands have open cracks
and sores that can cause bacterial infection, so when the subject went to wash their hands, the
data could be affected greatly because such a high amount of bacteria could already be on the
subjects hand and if the infected area was failed to be cleansed, but was swabbed, an extreme
amount of bacteria would be present (Secor). The research gathered helps the understanding of
the topic because it involves something that everybody can relate to which is patience. Not many
people want to check for the right temperature or wash for the best time even though it could
result in being as clean and healthy as possible.
This experiment can be used in many major corporations, small businesses and especially
hospitals. Hospitals can use this data to determine if the way that the staff is cleansing
themselves is effectively killing bacteria. It can also be used to see if the staffs hands are clean
enough to come in contact with the patients. Corporations can use this data in many ways.
Mainly so that cleanliness can be contained in the work environment and prevent diseases that
are spread through touch. Small businesses can use this data in the same way. Washing hands
efficiently can keep a small business clean. If one employee gets sick from not properly washing
their hands then the entire business can become sick and then company can suffer. Other
businesses can use this information to improve the products being sold. A hand soap company

Hauer Serraiocco 22

can recommend the best way to wash hands. Many of these examples are useful ways that this
experiment can be implemented into the real world.
A few lessons that can be derived from this experiment such as time management and
teamwork. Time management is extremely important during this process. It is important because
if the tests and the different sections start to pile up then numbers and results can be confused
and incorrectly used. If time is used wisely then the experiment will run smooth and with
minimal stress. Teamwork is also an important factor in the testing process. When two partners
are not working to the best of their abilities then the whole experiment can suffer for it.
However, when the partners are working at their best then the testing and recording can fly by. A
good team can make the difference in an experiment.

Acknowledgments
The researchers would like to thank the class of 9B for giving up their time to help test this
experiment. Other thanks go out to the families of the researchers for the constant support.

Works Cited
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<http://health.howstuffworks.com/skin-care/cleansing/basics/hand-washing2.htm>.

Bass, Pat, and Amy Finke, eds. "Hand washing: What is the best way to wash hands?" Banner
Health. N.p., n.d. Web. 16 May 2012.
<http://healthlibrary.bannerhealth.com/Library/Encyclopedia/85,P00628?PrinterFriendly
=true>.

Hauer Serraiocco 23

Eppes, Stephen C. "KidsHealth." Staph Infections. Apr. 2011. Web. 23 May 2012.
<http://kidshealth.org/parent/infections/bacterial_viral/staphylococcus.html>.

Fuller, Julia. "How Does Hand Soap Kill Bacteria?" EHow. Demand Media, 27 Jan. 2009. Web.
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<http://www.ehow.com/how-does_4740340_hand-soap-kill-bacteria.html>.

Hardy, Jay. "11 Hand washing Facts.. N.p., n.d. Web. 16 May 2012.
<http://www.hardydiagnostics.com/articles/hand-washing.pdf>.

Noorani, Shehzad. "The importance of hygiene." Unicef.org. N.p., n.d. Web. 24 May 2012.
<http://www.unicef.org/wash/index_hygiene.html>.