Jack Waitz

May 11, 2013

Science Slot B
Partner: Lauren Holtzman
Comparing Disinfectants
Purpose: To $ind out which disinfectants work best in preventing bacterial growth in our
school.
Hypothesis: If bacterial colonies are collected from the doorknob on the private bathroom,
and multiple disinfectants applied separately on each petri dish, then the most effective
disinfectant will leave the least surface area covered by bacteria on the petri plate, because
disinfectants disrupt normal bacterial functions to kill the bacteria and limit their growth,
and the better the disinfectant is at that, the less bacterial cells will be left.
Variables:
Independent Variable: type of disinfectant
Dependent Variable:The amount of Bacteria that grew.
Controlled variables: The amount of disinfectant, location in which the bacteria was
collected, heat of environment in which the bacteria grew, location of environment in
which the bacteria grew. Nutrient Agar for bacterial growth.
Control group: The bacteria on the petri plate without disinfectant applied
Materials:
Sharpie
3x Agar Petri Plates
Zep
Spic and Span
3x Fingers
Soap
Water
Doorknob of private bathroom in middle school hallway
Incubator
Dropper
Tape
Procedure:
1. Gathered petri plates and disinfectants. Labeled Petri Dishes with name and
disinfectant used.
2. Sterilized $inger by washing for 1 minute in hot water with soap.
3. Gathered bacteria from doorknob on private bathrooms pointer $inger.
4. Wiped bacteria on petri dish in a zigzag pattern.
5. Applied two drops of disinfectant to petri using dropper.
6. Shut and taped lid.
7. Repeated steps 3-6 with middle $inger and different disinfectant.

8. Repeated steps 3-4 and 6 with ring $inger, because the IV (disinfectant) is not
changed in a control group.
9. Put petri dishes in incubator.
10. Observed growth of bacteria on the petri dish once a day for three days, noting
number of colonies, approximate size, and total surface area covered.
Data
Comparison of Number of Bacterial Colonies, Size of the Colonies, and Total Surface Area
Covered between the Three Test Groups on Day 1--Chart 1
Type of
Disinfectant
Used

Number of
Colonies

Size of
Bacterial
Colonies

Surface
Area of
Petri Plate
Covered

Zep

Small to
Miniscule

0.5%

Spic and
Span

Small to
Miniscule

0.4%

Control

47

Small to
Miniscule

4.7%

Comparison of Number of Bacterial Colonies, Size of the Colonies, and Total Surface Area
Covered between the Three Test Groups on Day 2--Chart 2
Type of
Disinfectant
used

Number of
Colonies

Size of
Bacterial
Colonies

Surface
Area of
Petri Plate
Covered

Zep

medium to
medium
large

1.3%

Spic and
Span

medium to
medium
large

0.9%

Control

47

medium to
medium
large

11.5%

Comparison of Number of Bacterial Colonies, Size of the Colonies, and Total Surface Area
Covered between the Three Test Groups on Day 3--Chart 3

Type of
Disinfectant
used

Number
of
Colonies

Size of
Bacterial
Colonies

Surface
Area of
Petri Plate
Covered

Zep

medium
large to
large

2.6%

Spic and
Span

medium
large to
large

1.8%

Control

47

medium
large to
large

17.5%

Comparison of the Growth in Surface Area covered by Bacteria per Day--Chart 4

Type of
Disinfectant
used

Day 1 to
Day 2
Growth

Day 2 to
Day 3
Growth

Average
Growth

Zep

2.6x

2x

2.3x more
bacteria
per day

Spic and
Span

2.25x

2x

2.125x
more
bacteria per
day

Control

2.446808511x 1.52173913x 1.98427382

1x more
bacteria per
day

Average Daily 2.432269036 1.84057971x 2.1364246

Growth
7x
07x
Formulas Used:
1. Growth= Surface Area covered on Day/Amount of Surface Area Covered on Previous Day
2. Average Growth= ((Day 1 to 2 Growth)-(Day 2 to 3 Growth))/2,
3. Average Daily Growth= Growth Of (Zep Treated bacteria+Spic and Span Treated Bacteria
+ Control Group)/3

Control- Non treated

Zep

Spic and Span

Percent Surface Area covered per Day--Fig 1

Percent Surface area covered

30

22.5

15

7.5

0
Day 1

Day 2

Day 3

Day
Discussion
This experiment was an experiment done in order to see which disinfectants were most
effective at limiting bacterial growth. The original thought was that the best disinfectant
would leave the least surface area of the Agar Petri Plate covered, because Disinfectants are
meant to kill bacteria. However, the student did not know which disinfectant would be the
best. In the dish treated by Zep, there were 5 bacterial colonies (chart 1) to begin with, but
after that, number of bacterial colonies did not grow (charts 2-3). However, everyday, the
average amount more bacteria per day was 2.3x (Chart 4). The reason the number percent
covered went up was that though the amount of colonies stayed the same, the size of each
colony doubled each day. The case was almost the same for every dish. With the bacteria
treated by Spic and Span, there was originally 5 bacterial colonies (chart 1). That amount
did not change (charts 2-3) and the amount more surface area covered went up an average
of 2.125x each day, a mere .275x more than with Zep (chart 4), due to the change in size of
the colonies. On the control dish, the amount of bacterial colonies started at 47 (chart 1),
stayed the same throughout the experiment (charts 1-3). The average growth was
approximately 1.98x per day (chart 4). As shown, the only factor that changed in each dish
was the size of the colonies, causing the growth in size. Each dish each approximately
doubled each day, as the average growth per day was 2.1364624607x (chart 4). This
growth in size, not number, of the bacterial colonies becoming bigger can be attributed to
the genetic mutations of the bacteria in the colonies. After the $irst day, all the bacteria that
were not immune to the disinfectant were either killed or stopped from reproducing. The
only bacteria left were the bacteria that were immune to the disinfectant, and those
bacteria began reproducing, creating colonies around the afore-mentioned bacteria, but not
creating new colonies. No more bacteria entered the petri plate, so the only bacterial

colonies that could exist were the ones in which there was an originally immune bacteria.
This is one of the dangers of using disinfectants, that if one keeps using disinfectants on a
certain area, the only bacteria left will be the bacteria immune to everything, which will no
longer be able to kill be killed using disinfectants. When using disinfectants, one must kill
all bacteria to make sure this will not happen. Though one should not normally use
disinfectant for the reasons stated above, the most effective disinfectant was Spic and Span.
The most bacterial colonies on Spic and Span was 4, 1 less than Zep, and 43 less than the
non-treated control (charts 1-3). The total surface area covered was always less than the
other two groups ($ig 1), especially less than the control group, coming in with .8% less
covered than Zep, and 15.7% less than the control on the $inal day end (chart 3). Zep was
the less effective out of the two, though Zep still did much better than the control group,
showing that disinfectant is still better than nothing, the control. The control is much
needed in these experiments in drawing conclusions as the control shows what would
happen with no disinfectant. One can then compare what happens with disinfectant in the
different categories to see if the disinfectant actually did anything more successful than
normal. For this experiment, one important thing was to inoculate the bacteria from the
same site so that there would be the same amount of bacteria on the petri dish before the
disinfectant was applied. This limits sources of error. However, even when sterilized, a
$inger can not possibly not have bacteria already existing on the afore-mentioned $inger.
This is a source of error as some petri plates started out with more bacteria than others.
Also, the time that the $inger was rubbed against the doorknob varied, so some $ingers had
more bacteria on them. To solve these sources of error, one could use a sterile cotton swab
and wipe the doorknob for a set time, such as 5 seconds.
Conclusion
The hypothesis was approved, and the most effective disinfectant did kill the most bacteria.
The student did learn about using disinfectants and how disinfectants limit bacterial
growth.