BurginDrylie 1 Introduction Humans come into contact with millions of bacteria each day.

In fact, at any given time, a person could expect to find about 1, !! germs per s"uare centimeter on their hand. #$%a&e'Habit(). *hese bacteria should be removed from the hands through washing. +nfortunately, the ma,ority of all people use substandard practices while washing their hands. -omething as trivial as the temperature of the water or duration of time spent washing hands may not seem very important to one.s health, but these seemingly unimportant factors can critically affect the amount of bacteria removed from the hands through washing. Because washing hands is an effective way to prevent and reduce the spread of illness #$Hand /ashing'Infections(), humans should wor& as hard as possible to ma&e sure they are washing their hands properly. *here are many factors when it comes to washing hands properly, including temperature of the water and duration of time spent under the water #0arnum). *hese factors can directly affect the amount of bacteria left on hands after washing, which were the reslts we were trying to manipulate in our experiment. /e were curious about how much the factors of temperature and duration actually affect the amount of bacteria that grows on hands. /e set out to test the effect and interaction of these two specific factors on the growth of bacteria. /e hypothesi1ed that the warmest temperature for the longest amount of time would produce the least bacterial colonies. /e hoped to prove or disprove this hypothesis with our research experiment. 2ur first variable, temperature, did not have much definitive research as to whether or not it had a ma,or effect on the amount of bacteria &illed through hand washing. %ost of our research recommended warm or hot water when washing hands,

BurginDrylie 3 but we were not sure of the effect this variable would have. 4fter more extensive research, we found that the heat of hot water may have a better chance of &illing the bacteria on the hands, so we hypothesi1ed in favor of hot water. 4lthough research recommended that we use hot water to &ill bacteria, we had to find a temperature that would be safe and comfortable to use on human s&in. 2ur second variable, duration, has a more clear effect on the number of bacteria. It is simple to understand that the longer the hands are washed for, the more opportunity there is for the bacteria to be rinsed off #-lut1). /hen hands are being washed, the soap actually brea&s the pod5li&e structure that the bacteria use to stic& to the s&in. 4fter the pod is bro&en the water washes the bacteria off of the s&in. /ith a longer duration of time spent washing the hands, there is more time for the bacteria to be washed off the s&in. If we were to wash the hands for a shorter amount of time, there would be very little opportunity for the soap and water to do its ,ob with the bacteria on the hands. Because of our research, we decided to hypothesi1e in favor of warm water for the longest amount of time because that seemed li&e it was most li&ely to result in the least amount of bacterial growth. *o carry out our experiment, we had our test sub,ects wash their hands under a certain temperature for a specific amount of time and too& samples of the bacteria growing on their hands after they washed. /hile we were testing, there was a constant value, which was the non5antibacterial hand soap we used to wash the hands with. /e decided to use non5antibacterial hand soap because research suggested that there is actually very little benefit from washing with an actual antibacterial soap #$Hand

BurginDrylie 6 /ashing Do.s and Don.ts.() /e then incubated the 7etri dishes to see how much bacteria they yielded. Hopefully the results of our experiment will prove both helpful and convincing to people who are not sure of the correct temperature and duration for the most successful hand washing techni"ues. 4fter our hypothesis is proven or disproven, we will &now which levels are most effective for each of our variables, and our "uestions about proper hand washing will hopefully be answered.

BurginDrylie 8 7roblem -tatement 7roblem9 *o determine the effect of temperature of water and duration of exposure to water on the number of bacteria living on human hands. Hypothesis9 If the temperature of the water is at the highest temperature, 6:;<, and the hands are exposed to water for the longest amount of time, 6! seconds, then the least bacteria will grow from the collected sample. Data %easured9 *he independent variables in this experiment were the temperature of the water, measured in degrees <elsius #;<), and the amount of time that the hands were exposed to the water, measured in seconds. *he dependent variable in this experiment was the number of bacterial colonies grown from the collected samples. *he data analysis used was a two5factor Design of =xperiment #D2=.)

BurginDrylie =xperimental Design %aterials9 1!! x 1 mm 7etri Dishes #>!) ?utrient 4gar *est *ubes #6) -terile Distilled /ater <ulture <ollectors #>!) *I5nspire <alculator Hot 7late -terile -tirring %agnet %etal *ransfer @oop Bunsen Burner 7rocedures9 <B=4*I?C CB2+7Temperature Low (-) Standard High (+) 4 C 21 C 38 C Duration Standard 2 !

1 @ 0las& Incubator at 6A; < -harpie %ar&er *hermometer -oftsoap Brand Hand -oap /ater -ource 1 m@ Dropper Hot %itt Bunsen Burner @ighter -topwatch

Low (-) 1 !

High (+) 3 !

B4?D2%IDI?C *BI4@ ?+%B=B1. 3. 6. -et the random integer function on the *I5nspire calculator to #1, 36) and hit enter. Eeep trac& of the order in which the numbers were chosen. -ort them into groups, with the first test number being #F,F), the second being #F,5) the third being #5,F), the fourth being #5,5) and the fifth being the standard. <ontinue this pattern until all test sub,ects have been assigned a group.

8.

4C4B 7B=74B4*I2? 1. 4dd 36 g of agar medium in to 1 @ of purified water in the 1 @ flas&.

BurginDrylie > 3. 6. 8. 4dd stirring magnet to the flas&. 7lace flas& on hot plate, and turn on the plate. Heat while stirring and boil for one minute to dissolve completely, being sure to watch that the agar does not bubble over. *urn off heat. Bemove flas& from hot plate, using a hot mitt.

. >.

=G7=BI%=?* 1. 3. 7repare agar using procedures above. 7repare 7etri dishes by pouring nutrient agar to cover the bottom of the plates, and let the agar cool for approximately 1 minutes to harden. 0ill one of the test tubes with sterile distilled water. +sing the dropper, place 1. m@ of sterile distilled water in the second and third test tubes. +sing the -harpie mar&er, label the first 7etri dish with the number of the test sub,ect and the group the sub,ect was placed in. +se the thermometer to determine the temperature of the water. Baise the water temperature by turning the hot &nob, and &eep measuring the temperature until the thermometer shows 6:;<. Have the first person wash their hands with one s"uirt of the non5antibacterial soap for thirty seconds under the hot water, using the stopwatch to &eep trac& of how long they have been washing their hands. 2nce that is done, ta&e the full test tube and pour the sterile water over the hand of the person.

6. 8.

>.

A.

:.

BurginDrylie A H. *a&e a culture collector and swab the person.s hand all over, ma&ing sure to get between fingers and in other crevices. 7lace the culture collector in the second test tube. 4fter letting the culture collector soa& in the 1. m@ of water, remove the culture collector from the water. +se the Bunsen burner lighter to light the Bunsen burner. 0lame the transfer loop till the loop turns red. Drop the transfer loop into the test tube that the culture collector was previously removed from #second test tube). %ove the transfer loop from the second test tube to the third test tube. Bemove the lid from the 7etri dish and pour 1. m@ of water from the test tube into the dish. Invert the plate by flipping the dish so that the cover is face down and the bac& of the dish is facing up. 7lace the plate into the incubator. -et the incubator to 6A; <. Bepeat these procedures using the correct temperature and duration for the other groups. 4fter three days, remove 7etri dishes from incubator. <ount colonies and record trial number, group, and number of colonies.

1!.

11. 13. 16.

18. 1 .

1>.

1A. 1:. 1H.

3!. 31.

BurginDrylie :

0igure 1. Bacterial Dilution 0igure 1 above is a diagram showing the procedures ta&en to dilute the bacteria. *he first step was placing the culture collector in the first test tube. *hen the collector was removed and the transfer loop was placed in the test tube. *he loop was moved from the first test tube to the second tube, then the tube was poured into the 7etri dish.

BurginDrylie H Data and 2bservations Data9 *able 1 ?umber of <olonies Crown 7er 7etri Dish
Tria% 1 'un! (+( +) (+( -) (-( +) (-( -) Standard (+( +) (+( -) (-( +) (-( -) Standard (+( +) (+( -) (-( +) (-( -) Standard (+( +) (+( -) (-( +) (-( -) "um#er o$ Co%onie! Tria% 2 &rder 'un! 'e!u%t 12 (+( +) 2) 8 (+( -) + 1* (-( +) +) 2 (-( -) 24 1 Standard 1) 13 (+( +) 2 , (+( -) , 1+ (-( +) ,) 3 (-( -) 23 4 Standard 13) 14 (+( +) 3) 1 (+( -) 4 18 (-( +) +) ) (-( -) 1* + Standard , 1) (+( +) 4 11 (+( -) 8 1, (-( +) ,) * (-( -) 1, Tria% 3 'un! (+( +) (+( -) (-( +) (-( -) Standard (+( +) (+( -) (-( +) (-( -) Standard (+( +) (+( -) (-( +) (-( -) Standard (+( +) (+( -) (-( +) (-( -)

&rder 12 2 8 1* 1 13 3 , 1+ 4 14 ) 1 18 + 1) * 11 1,

'e!u%t 2 4 ) 3 1 * 2 + 2) 14 2 * , 18 11) 12 * , 1,

&rder 1* 2 12 8 1 1+ 3 13 , 4 18 ) 14 1 + 1, * 1) 11

'e!u%t 2 +) 28 14 1 3) ) 2, 13) 2 1 , 2 1)) 3 1) 8 21

*able 1 above shows the raw data collected during the experiment. In the results column, it is shown how many colonies were grown in that specific 7etri dish. *he table shows the order in which the plates were tested and the combination of the levels of the two factors. *able 3 Design of =xperiment Ialues
4 Temperature (C) Standard 21 + 38 1 Duration (!) Standard 2 + 3

*able 3 above shows the factors that went into the experiment, temperature and duration. It gives the low, standard, and high values for the factor of temperature in degrees <elsius. It also gives the low, standard, and high values for the factor of duration

BurginDrylie 1! in seconds. *his table was used as a reference in the experiment to be sure the right values were being used. *o select the levels for each factor, research was used which showed the ideal temperature and duration of hand washing, which was the standard. *he lows and highs were decided by going up and down by an e"ual increment 2bservations9 *able 6 *rial 2bservations
(+(+) (+(-) Tria% 1 (-(+) (-(-) (+(+) (+(-) Tria% 2 (-(+) (-(-) (+(+) (+(-) Tria% 3 (-(+) (-(-) Thi! group grew the %ea!t amount o$ -o%onie!( with an a.erage o$ ten -o%onie! per p%ate/ 0! -ompared to the (+(+) and (-(-) group!( thi! group grew an amount o$ -o%onie! !omewhere in #etween/ 1rew !%ight%2 more -o%onie! than the (+(-) group/ 1rew the mo!t -o%onie! out o$ an2 o$ the group!/ Thi! group a%!o grew the %ea!t amount o$ -o%onie! on a.erage a$ter te!ting the !e-ond tria%/ 0! in tria% 1( thi! group grew -o%onie! !omewhere in the midd%e o$ the %ow group and the high group/ Thi! group3! -o%onie! were %arger and had a higher amount o$ tho!e in the !ame group $rom tria% 1/ Thi! group a%!o grew on a.erage the highe!t amount o$ -o%onie! out o$ an2 group in thi! tria%/ Co%onie! !ti%% %oo4 norma%/ 5%ate 1 ha! no -o%onie! though it wa! gi.en the !ame treatment/ There wa! more o$ a !igni$i-ant di$$eren-e #etween thi! group and the (-(+) group in thi! tria%/ 5%ate 11 had what %oo4ed %i4e $i%m -o.ering it( #ut the re!t %oo4ed %i4e the2 were e6pe-ted to/ Sti%% growing the mo!t #a-teria/ 5%ate 14 grew wa2 more than e6pe-ted/

*able 6 gives information that was collected while experimenting. *he table gives observations of the four separate groups as a whole instead of each individual plate. /ithin the observations, trials and groups are compared to one another and compared to themselves. 4lso there are a few individual observations of a few plates that seemed to be outliers in the experiment. /e followed our procedures exactly throughout the experiment, so procedures were not noted in the observations table.

BurginDrylie 11

-ingle colony

Croup of colonies

0igure 3. =xample of <olonies 0igure 3 shows examples of what the colonies loo&ed li&e. *his specific plate came out of one of the #F,F) tests. *he typical colony loo&s li&e one of the single dots in the plate. 4lso, there are several groups of colonies in this specific test. *his also held true with the other tests throughout the other groups.

BurginDrylie 13 Data 4nalysis and Interpretation Data 4nalysis9 *able 6 Design of =xperiment Ialues
4 Temperature (C) Standard 21 + 38 1 Duration (!) Standard 2 + 3

*able 6 above is a &ey showing the values assigned for each variable at the low, standard, and high values. *hese values were assigned after extensive research showing the recommended level, which was the standard. *he lows and highs were found by raising and lowering each standard by a specific number of units. *able 8 Besults 0rom 4ll *rials
D&7 1 2 3 081 (+(+) 1 3 1) 18/3 (+(-) 4) + * )8/3 "um#er o$ Co%onie! (-(+) (-(-) +) 23 8) 2 ) +) 24) +8/3 22*/+ Standard 118/3 12) 143/3 128/,

*able 8 above is a table showing the results of the experiment for each trial. *he results are shown for each levelJvariable group. *he standard results are also listed. *able =ffect of *emperature
Temperature + 4C 38C 22*/*+ 18/3 +8/ 3 )8/3 0.g91)2/) 0.g938/3

BurginDrylie 16

*able shows the average number of colonies for the D2= groups in which temperature was at the low level and high level. *he averages of the number of bacterial colonies, were graphed in 0igure 3 below. Effect of Temperature
24 / Number of Colonies 2 1)2/) /

1* / 12 / 8 / 4 / / 38/3

-1 Temperature

0igure 6. =ffect of *emperature on ?umber of <olonies *able > =ffect of Duration

Duration + 1 ! 3 ! 22*/*+ 18/3 )8/3 +8/3 0.g9142/) 0.g948/3

*able > above shows the average number of colonies for the D2= groups in which duration was short and long. *he averages of the bacterial colonies were used in the graph in 0igure 6.

BurginDrylie 18

Effect of Duration
24 / Number of Colonies 2 142/) /

1* / 12 / 8 / 4 / / 48/3

-1 Duration

0igure 8. =ffect of Duration on ?umber of <olonies *able A Interaction =ffect

Temperature Line Segment So%id Line Segment Dotted Duration +8/3 + 22*/+ )8/3 + 18/3

*able A above shows the interaction effect of the two variables9 temperature and duration. It shows the points graphed in the solid and dotted line segments, which in turn allowed the interaction effect of temperature and duration to be found.

BurginDrylie 1

Interaction Effect: Duration and Temperature

22*/+ 24 / 2 /

Number of Colonies

Duration (-)

1* / 12 / +8/3 8 / 4 / / -1 Temperature 1

Duration (+)

)8/3 18/3

0igure . Interaction =ffect of Duration and *emperature *able : ?ine -tandards

1 14 11) "um#er o$ Co%onie! 1) 13) , 14 13) 1))

*able A above shows the number of colonies grown from each of the standard trials.
Standards
24 Number of Colonies 2 1* 12 8 4 1 3 ) Trial Number + ,

0igure >. -tandards -catter 7lot

BurginDrylie 1> Interpretation9 0igure 6 shows the effect that temperature had on the number of bacterial colonies in the experiment. /hen the temperature of the water was at the low value, the sample plates grew 1 3.3 colonies on average. /hen the temperature was at the high value, the sample plates grew 6:.6 colonies on average. *emperature had a negative effect on the colonies, because the slope of the main effect line segment was negative. *he effect of temperature, or the change from low to high, was 5118.3 colonies. 2n average, as the temperature increased, the number of colonies grown on the sample plates decreased by 118.3. *his effect probably occurred because the hot water had an easier time removing bacteria from the hands. 4lthough we could not find much definitive research as to whether or not temperature had a ma,or effect on the bacteria, all research seemed to thin& that the warmer water may be more effective. 2ur research seemed to prove this. 0igure 8 shows the effect that duration had on the number of bacterial colonies grown throughout the experiment. /hen the duration of time spent washing hands was at the low value, the sample plates grew 183. colonies on average, but when the duration of time was at the high value, the sample plates grew 8:.6 colonies on average. Duration had a negative effect on the number of colonies because the slope of the main effect line segment was negative. *he effect of duration, or the change from low to high, was 5H8.3 colonies. 2n average, as the duration increased, the number of colonies decreased by H8.3. *his effect probably too& place because the longer the hands were exposed to the water, the more opportunities there were for the bacteria to be rinsed off of the hands. 0igure shows the interaction that too& place between both main effect factors.

*he graph is implying that there is an interaction because the line segments have different

BurginDrylie 1A slopes, and may have intersected if we had a larger range of data in our experiment. *he two factors seemed to wor& with each other because the slope of the dotted line and the slope of the solid line are both negative. *he slope of the line Duration #F) minus the slope of the line Duration #5) gives the effect #duration vs. temperature) 8.3, so the interaction effect of duration and temperature is 8.3. 0igure > shows all the standard results from the experiment. *he standards show that the experiment was done correctly, because there was a somewhat hori1ontal linear relationship between the points on the scatter plot.

BurginDrylie 1: <onclusion 4fter carrying out this experiment, it was concluded that the original hypothesis was accepted. *he original hypothesis was that if the temperature and duration were both at the highest level, the least bacteria would grow from the selected sample. *he results of the experiment went along with this because both temperature and duration had a negative effect on the number of colonies grown in the experiment. In this experiment, it was planned to find the ideal values of temperature and duration of water exposure while washing hands. It had been discovered that most people do not &now the correct procedures for washing hands, and may not even &now the benefits of washing correctly. *his experiment, was carried out to prove and convince that washing hands correctly can remove infectious bacteria from human hands. *he different levels were tested by washing the test sub,ects. hands with a certain temperature of water for a certain amount of time. 4fter they washed, samples were ta&en and bacteria were grown in 7etri dishes. *he results of the experiment showed that washing hands with warmer temperatures for longer periods of time removed the most bacteria. *here was also an interaction between the two variables. It was concluded that they wor&ed together to remove the bacteria from the hands. It can not be &nown for certain why these results were attained, but it can be inferred. *he temperature variable probably wor&ed negatively with the colonies, because the hot water had a better chance of &illing the bacteria then the cold water. *he duration variable wor&ed negatively with the colonies because the longer the hands were washed for, the more opportunities there were for bacteria to be removed from the hands.

BurginDrylie 1H *here were a few design flaws throughout this experiment. 2ne of the ma,or design flaws was corrected during the pre5tests. 0or the pre5test trials, the bacteria samples were not diluted before pouring them into the plates. *his resulted in bacterial growth with so many colonies that it was nearly impossible to count the number of colonies. *his flaw was corrected before the actual trials started, so a manageable number of bacterial colonies were able to be reached. 4nother design flaw that was encountered was that there was difficulty finding accurate temperatures. *his design flaw was by using a glass thermometer as opposed to the original metal thermometer we used in our pre5tests. ?o &nown errors were made while carrying out the procedures. 2nce the design flaws described above were corrected, the procedures were followed directly. If more research was conducted to improve the understanding of this topic, the experiment may have been done differently. 4n ideal experiment would have had more than 1H test sub,ects, but this had to be done on a smaller scale test because of the limited resources provided. If the experiment itself was modified, the scientists may consider changing the levels of the variables to further prove the results of this experiment. *he research done enhanced the understanding of this topic because it helps to show how the temperature and duration of the water affects the amount of bacteria that are removed from hands while washing. *he results of this experiment could be used to help the community. Iery few people actually &now the ideal temperature or duration of water to wash their hands in. /ith the information and data collected in this experiment, it was proved that hands should be washed in warm water for at least 6! seconds. If people were to wash their

BurginDrylie 3! hands at these ideal levels, more bacteria could be removed from the hands. *hese results can be used specifically in medical or food preparation environments, because it is vitally important to &eep bacterial levels as low as possible in these areas. -ince washing hands correctly is one of the most important factors in avoiding and reducing the spread of illness or infection, the results of this experiment can be used to &eep people healthy. %any things were learned throughout the course of this experiment. %ost importantly, the benefits of extensive research were discovered before beginning the actual experiment. It was also discovered that it was imperative to learn a lot about this topic before testing the variables could be done. *hrough this, it was discovered how to go through the correct procedures to carry out a biological experiment. Enowledge was gained about bacteria growing on human s&in, and &nowledge about how to arrange the results of the experiment in a way that ma&es sense both scientifically and mathematically was also gained.