Testing for the Presence of E.

Coli, Staphylococcus and Streptococcus Through Casual

Catch Urine Samples from Males and Females
Jonathan Smith

Purpose
The purpose of this experiment is to test for the presence of E. Coli, Staphylococcus and
Streptococcus in male ad female casual catch urine samples.
Introduction
Urinalysis is a way of testing to examine the health of the urogenital system. This system
is comprised of two kidneys, two ureters, one bladder and a urethra. The health of this
system is vital to removing harmful waste from your body and reabsorbing nutrients
before they are expelled. This system helps the body maintain homeostasis. A common
problem that the urogenital system encounters is a Urinary Tract Infection.
These infections are most common in women and are tested using either casual or clean
urine samples. Casual catch samples are collected after the genital area has been wiped
with an alcohol pad. Casual catch samples are collected without any prep of the genital
area. In this experiment, casual catch samples were used, meaning that the results will not
show if a UTI, rather they will show the bacteria that can be present in a urinary tract
system.
This experiment will be testing to show the bacteria that may be entering or present in our
body from the urinary tract system. This experiment will be testing to find the presence of
E. Coli, Staphylococcus and Streptococcus. E. Coli is naturally found inside of intestines
and produces vitamin k and b-complex. E. Coli will grow on the purple agar, which is
called Eosin Methylene Blue and is gram-negative. Gram-negative agar will only grow
gram-negative bacteria, meaning that E. Coli is the only gram-negative bacteria this
experiment hopes to find. If E. Coli is present then it will look metallic green.
Staphylococcus is the bacteria found inside of the bodys natural flora and is found on the
skin as well as on mucous membranes and the vagina. Streptococcus is also found on the
body naturally and is found on the same places as the Staphylococcus. Streptococcus and
Staphylococcus will both grow on the Phenylethanol agar. This agar is gram-positive and
both of these bacteria only grow on the gram-positive agar. Both of the bacteria will
appear white, however staphylococcus will react with hydrogen peroxide, creating
bubbles and streptococcus will not cause a reaction.

Hypothesis
1. If Escherichia coli is present in the urine sample, then green bacteria will grow on
the EMB culture plate.
2. If Streptococcus is present in the urine sample, then white bacteria will grow on
the EMB culture plate.
3. If Staphylococcus is present in the urine sample, then white bacteria will grow on
the EMB culture plate and will react to hydrogen peroxide.
4. If the urine sample is from a female, then more bacteria will be found in the EMB
culture plate than in the males culture plate.
Materials and Equipment

Cotton Balls
Rubbing Alcohol
1 Petri Dish
Q-Tips
Sharpie
Tongs
Refrigerator
Incubator
Phenylethanol Agar
Eosin Methylene Blue Agar
Hydrogen Peroxide
Sample Cup Full of Urine

Methods
1. Use cotton ball drenched in alcohol to sterilize your work area
2. Use sharpie to label the four sections of the petri dish. Two sections should be
labeled positive and the other two negative
3. Place the petri dish inside of the sterile work area
4. Heat agars until they become a liquid inside of a test tube
5. Use tongs to grab test tube with one kind of agar
6. Use lighter to heat end of test tube with agar to clear of any bacteria
7. Pour it into a positive and a negative section of the petri dish
8. Use tongs to grab the other kind of agar and pour it into the remaining sections of
the petri dish, a positive and a negative section
9. Place petri dish in refrigerator until the agar has solidified again
10. Let agar sit for 24 hours in the refrigerator
11. Collect urine sample in sterile cup
12. Dip Q-tip in the urine sample until one end is completely covered

13. Gently drag the urine soaked end of the Q-tip in a zigzag pattern across the top the
agar in both positive sections of the petri dish
14. Safely dispose Q-tip, urine sample and other contaminated objects
15. Cover petri dish with lid and place in incubator, set at 98 degrees Fahrenheit, for
24 hours
16. After 24 hours, open incubator and remove petri dish
17. Open lid and check for bacteria
18. If white bacteria are present, isolate a small area and use hydrogen peroxide to
test for Staph.
19. Place lid on petri dish and place back inside of incubator set at 98 degrees
Fahrenheit for 24 hours
20. Record results on table
21. After 24 hours, open incubator and remove petri dish
22. Repeat steps 14-16
Results and Analysis

Presence of Bacteria in Urine Samples by Gender

Male +
Female +

E. Coli

Strep.

Staph.

After analyzing the results from the experiment, it can be concluded that more males
samples contained bacteria than female samples. Staphylococcus was the most common
bacteria found in the samples as it was found in 29% of the male samples. It was found in
18% of the female samples. Streptococcus was found in 14% of the male samples and
was not found in any female samples. E. Coli was not found in any of the male samples
and was found in 5% of the female samples.
Male +
E. Coli
Strep.
Staph.

0
4
10

Female
+
1
0
3

Discussion

The purpose of this experiment was to test for the presence of three different bacteria that
could be inside of urine samples. Bacteria are the main cause for urinary tract infections,
which are most common in women. The results of this experiment are not meant to show
if the subject has a UTI, rather it was meant to show the bacteria that might be present in
a urinary tract infection. Hypothesis number one is rejected for 100% the male urine
samples as green E. coli bacteria were not found on the EMB culture plate. This can be
expected because the male urethra is longer than the females, thus causing a longer
distance for the bacteria to travel. Hypothesis one can be accepted for 5% of the female
samples and is rejected for the remaining 95% of the female urine samples. This is to be
expected because the female urethra is shorter and touches more places allowing more
bacteria to grow. Hypothesis number two is accepted for 14% of the male urine samples
as white bacteria were found on the EMB culture plate. The hypothesis is rejected for
86% of the male urine samples due to the lack of white bacteria on the EMB culture
plate. Hypothesis number two is rejected for 100% of the female samples due to the lack
of white bacteria on the EMB culture plates. Hypothesis number three is accepted for
29% of the male urine samples as that amount had white bacteria that were found on the
EMB culture plate that reacted with hydrogen peroxide. This hypothesis was rejected for
71% of the remaining male samples. Hypothesis number three is accepted for 18% of the
female samples and was rejected for 82% of the female samples. Hypothesis number four
is rejected for 100% of the samples. While the female samples did show bacteria, a larger
amount of male samples tested positive for bacteria. This is unexpected because the male
urethra is longer than the females, meaning that the bacteria have a longer distance to
travel before it can actually thrive inside the body. In this experiment, the most common
bacteria were Streptococcus. This experiment found Streptococcus inside more male
urine samples than female samples. This is to be expected because Streptococcus is found
naturally on the human bodys skin and inside the mucus membranes. It is surprising that
it was found on more male samples than female as it is also found naturally on the
vagina. The least common bacteria found were the Escherichia coli. It was only found on
one female sample and was not present on any of the male samples. This is surprising

because E. coli is a producer of vitamins k and b. This lab should be considered reliable
for a number of reasons. This experiment used 61 different casual catch urine samples,
which covered a variety of subjects. Only two samples out of 61 were contaminated,
bacteria on the negative side of the EMB culture plate, suggesting that the remaining
samples were prepared in the proper way. The only adjustment that should be made if this
experiment were to be repeated would be to let the EMB culture plates sit upright for at
least eight hours before flipping them and placing inside of the incubator. This would
allow the bacteria to spread evenly and for more positive results. In conclusion, this
experiment showed accurately showed the bacteria that could be present inside casual
catch urine samples.