Bacteria Bustin:

The Effects of Antibiotics on Bacteria

Project #121
4/7/2011

Abstract

I. INTRODUCTION
Our science fair project is an experiment revolving around the use of antibiotics to fight
off bacteria. The first bacteria we use is E. coli. Escherichia coli also known as E. coli is a
common bacterium found in the intestinal tract of warm blooded animals and in soil and water. It
was discovered in 1885 by Theodor Escherich. In 1892 a scientist proposed that E. coli could be

used to detect fecal matter in water supplies due to the fact that it is not too common elsewhere,
and is distinguishable based on its ability to change glucose into lactose. It is for the most part a
very helpful organism because it creates Vitamin K for the organism in which it dwells. However,
there are some more dangerous strains of the bacteria that can cause diarrhea or other
gastrointestinal complications. E.coli does not generally survive well outside of the human
intestines. E. coli is also a substance used in the indication of carthogens in humans.
E. coli belongs to the variety of bacteria generally deemed as coliform. This means that it is a
rod-shaped bacteria that changes glucose to lactose to gas. E. coli is susceptible to many
antibiotics including amoxicillin, penicillin, cephalosporins, carbepenens, aztreonam,
trimethoprim-sulfamethoxazole, ciproflaxacin, nitrofurantoin, and aminoglycosides. It has in
recent years developed a resistance to certain antibiotics.
Also a vital part of our experiment, bacillus cereus is a peculiar specimen. Discovered in 1887
by Franklind and Franklind, it is a bacterium that forms spores, and is a cause of food poisoning.
Bacillus cereus (commonly shortened to b. cereus), produces a toxin specific to the intestines,
that is very often affiliated with starchy foods such as fried or boiled rice, mashed potatoes, and
pasta. While having this bad reputation, it also assists in fighting off Salmonella, improving the
host animals growth and food safety. It is of the same genus as bacillus anthracis, the bacteria
that causes anthrax, which is a disease that infects the skin, lungs, or gastrointestinal tract.
In addition to bacteria, our experiment involves antibiotics. The following antibiotics are all
bacteria-killing medicines. Penicillin, probably the most common antibiotic, kills infections
caused by bacteria. Streptomycin is used to treat turberculosis and other sensitive bacteria.
Tetracycline, the third antibiotic, like penicillin, also treats infections such as acne, urinary tract
infections, etc.
Penicillin kills bacteria by breaking down the cell wall, making it frail and easy to destroy.
Penicillin was the first medicine of its kind, discovered by the Scotland-native Sir Alexander
Fleming in 1928 at St. Marys Medical Hospital in London. It eradicates many bacteria, even
though most grew resistant today.
Streptomycin is used for treating tuberculosis. It was first discovered in October of 1943 by a
student at Rutgers University, Albert Schatz. Along with this antibiotic, several others were found
in the lab. Streptomycin also kills other sensitive bacteria by terminating the necessary proteins
for the bacteria to grow and function.
Tetracycline kills several different kinds of bacteria, including acne and other skin infections,
urinary tract infections, and pneumonia and other respiratory infections. It prevents amino acids
from reaching the peptide chain, which is needed for the bacteria to spread and increase.
Tetracycline also breaks down scar tissue and produces healthy skin growth which helps scars
to become less noticeable. It was discovered by Benjamin Minge Duggar in 1948.
In addition to these antibiotics there are also ampicillin, chloramphenicol, and
gentamycin. Ampicillin is a semi synthetic form of the antibiotic, penicillin. The difference
between the two antibiotics is that the ampicillin has one extra amino group. The extra amino
group makes ampicillin effective against certain organisms, such as gram negative organisms.
The antibiotic has been used since 1961. It treats infections of the ear, skin, lungs, and urinary
tract; along with bronchitis. Ampicillin does not treat the flu or other viral infections. This
antibiotic also has side affects such as upset stomach, diarrhea, mild skin rash, and vomiting.
Chloramphenicol is an antibiotic that, like all antibiotics, helps to kill bacteria.

Chloramphenicol is mainly used after other antibiotics have failed to eliminate the bacteria.
Chloramphenicol also treats against dangerous illnesses such as plague, tularemia, and anthrax
of the skin and mouth. However this antibiotic has its side affects; for example, nausea,
vomiting, diarrhea, depression, mouth sores, depression, confusion, and headache.
Gentamycin is an antibiotic used to treat severe infections in patients in hospitals, nursing
homes, or other health care centers. This antibiotic started to be used around 40 years ago and
it has proven to be very effective in treating infections. Doctors have found, however, that
gentamycin has an extreme effect on the inner ear, causing permanent balance impairment and
bouncing vision.

II. PROCEDURE
Problem: What kind of antibiotic works best to kill bacteria?
Experimental Design:
Independent Variable(s): Antibiotics.
Dependent Variable: Growth of bacteria.
Controlled Variables: Experiment environment (i.e. lighting, temperature, etc.),
experimenters etc.
Control Group: Bacteria with no antibiotics.
Hypothesis: If the bacteria E. coli had the antibiotic ampicillin introduced, then it would be the
most diminished when compared to other antibiotics present in the experiment. If the bacteria B.
cereus were to be exposed to the antibiotic chloramphenicol, then it would be the most
diminished when compared to other antibiotics present in the experiment.
Materials:
(1) Bacilius cereus Broth Culture
(1) Escherichia coil Broth Culture
(4) (Disks Each) Antibiotic Disk Magazines (ampicillin, chloramphenicol,
gentamycin, penicillin, streptomycin, and tetracycline)
(4) Nutrient Agar (dishes)
(1) Control Disk (white)
(1) Metal Forceps
(1) Flame
(1) Incubator
(1) Autoclave Disposal Bag
(4) White Lab Coats
(4) Eye Goggles
(4) Pairs of Rubber Gloves
(1) Roll of Paper Towels
(1) Container of Anti-Bacterial Wipes
(1) Container of Clorox Bleach

(1) Roll of Tape

(1) Trash Bag
Procedure:
1.
Prepared the materials for the procedure.
1) Labeled Petri dishes: B. cereus 1, B. cereus 2, E. coli 1, E. coli 2.
2) Made key for antibiotic placement.
3) Placed marker on Petri dishes to indicate placement of antibiotics.
2. Disinfected all surfaces.
3. Washed hands and put on gloves, lab coats, and goggles.
4. Carefully poured bacteria culture of B. cereus onto the nutrient agar plates/dishes.
5. Carefully poured bacteria culture of E. coli onto the nutrient agar plates/dishes.
6. Disposed of bacterial containers into autoclave disposable bag.
7. Flamed forceps; allowed a few moments to cool.
8. Using flamed, cooled pair of forceps, removed one antibiotic disk (penicillin) from its package
and placed it onto the agar of the B. cereus culture disk. Used the previously made antibiotic
placement indicator for a guide.
9. Flamed forceps again after placing down the antibiotic disk.
10. Followed steps 6-8 for each antibiotic.
11. Followed steps 6-9 for E. coli.
12. Put bacteria into incubator.
13. Waited 24 hours and recorded diameter of no bacteria around each antibiotic disk.
14. Put bacteria back into the incubator.
15. Waited another 24 hours and again recorded the diameter of no bacteria surrounding the
antibiotic disks.
16. Put bacteria back into the incubator.
17. Waited another 24 hours and again recorded the diameter of no bacteria surrounding the
antibiotic disks.
18. Put Clorox Bleach into the Petri dishes, and taped the lids shut.
19. Placed the Petri dishes into a trash bag and tied it shut.
Clean up was steps 18 and 19. As a safety precaution we always wore goggles, a lab coat, and
gloves. Once set up with the antibiotics, we did not open the Petri dishes, save to kill the
bacteria with the bleach. We washed the surfaces we were using before and after each time we
took our measurements, and we washed our hands.

III. DATA
Tabl
e1

Bacterial Growth Chart Trial 1

Growth of the Diameter of
Cleared Area

Disk
Anti
bioti
c

Code

24 Hours

48
Hours

72
Hours

B. cereus

Amp
icillin

Chlo
ram
phen
icol

Gent
amy
cin

Peni
cillin

Stre
ptom
ycin

AM10

C30

GN10

P10

S10

Tetra
cycli
ne

T30

Tabl
e2

Bacterial Growth Chart Trial 2

0 mm.

0 mm.

0 mm.

0 mm.

0 mm.

0 mm.

E
.
c
o
l
i

B.
cereu
s

E
.
c
o
l
i

B.
cereu
s

E
.
c
o
l
i

m
m
.

m
m
.

m
m
.

1 mm.

3 mm.

m
m
.

m
m
.

m
m
.

1 mm.

0 mm.

m
m
.

m
m
.

m
m
.

1 mm.

0 mm.

m
m
.

m
m
.

m
m
.

1 mm.

2 mm.

m
m
.

m
m
.

m
m
.

1 mm.

2 mm.

m
m
.

m
m
.

m
m
.

1 mm.

0 mm.

Growth of the Diameter of

Cleared Area

Disk
Anti
bioti
c

Code

B. cereus

Amp
icillin

Chlo
ram
phen
icol

Gent
amy
cin

Peni
cillin

Stre
ptom
ycin

Tetra
cycli
ne

AM10

C30

GN10

P10

S10

T30

48
Hours

24 Hours

0 mm.

0 mm.

0 mm.

0 mm.

0 mm.

0 mm.

E
.
c
o
l
i

B.
cereu
s

72
Hours
E
.
c
o
l
i

B.
cereu
s

E
.
c
o
l
i

m
m
.

m
m
.

m
m
.

2 mm.

2 mm.

m
m
.

m
m
.

m
m
.

0 mm.

0 mm.

m
m
.

m
m
.

m
m
.

0 mm.

0 mm.

m
m
.

m
m
.

m
m
.

0 mm.

0 mm.

m
m
.

m
m
.

m
m
.

0 mm.

0 mm.

m
m
.

m
m
.

m
m
.

0 mm.

0 mm.

IV. GRAPHS
V. ANALYSIS
Based upon the graphs and charts, it would appear that ampicillin worked the best
against the bacteria. Tetracycline also worked well. Out of all of the bacteria, B. cereus (Trial 1)
was the least strong, so the antibiotics worked best on it. Other than ampicillin and tetracycline,
penicillin and streptomycin had a more major effect on B. cereus (Trial 1) than the other
antibiotics. Overall, chloramphenicol and gentamycin worked the least well.

VI. CONCLUSION
Based on our results, half of our hypothesis was correct. Ampicillin did work best overall,
we predicted that it would work best on E. coli, which it did. Our hypothesis was also wrong in
stating that chloramphenicol would work best on B. cereus. In fact, quite the contrary was true:
chloramphenicol along with gentamycin worked the least. Some things that may have affected
our experiment would be the fact that the incubator did not stay at a constant temperature, but
did, in fact, fluctuate on a daily basis. In addition to this, we did not flame the forceps after each
use. Also, the agar on B. cereus (Trial 1) was scratched when tetracycline was incorrectly
placed and was attempted to be moved. Also, the first time we examined the bacteria, we
observed it by holding it up to sunlight. The next time, however, we used a 100 watt bulb. More
trials without these errors would probably be good to do.

VII. ACKNOWLEDGEMENTS
We would like to thank our biology teacher, Mrs. Lesutis, who not only got the material
necessary for the project for us, but also helped us complete the experiment. We would also
like to thank Mrs. McClennan who showed us how to properly dispose of the bacteria, and who

provided the agar dishes, the incubator, and the space we worked in. Finally, we would like to
thank our parents, for getting us to school early to work on this, and for putting up with our
various antics.

VIII. BIBLIOGRAPHY
"Bacteriophages Offer Alternative to Antibiotics." Todays Science On File: n. pag. Todays
Science. Facts On File News Services, 31 Aug. 2000. Web. 4 Mar. 2011.
<http://www.2facts.com/article/s0802500>.
Feng, Peter, Michael A. Grant, Stephen D. Weagant. (2002, September). BAM: Enumeration of
Escherichia coli and the Coliform Bacteria. U.S. Food and Drug Administration. Retrieved
March 2, 2011, from
http://www.fda.gov/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalyticalManual
BAM/ucm064948.htm#authors
Newton, D. E. Food-borne illnesses and irradiation of foods. In Science online. Retrieved from
http://www.fofweb.com/activelink2.asp?
ItemID=WE40&SID=5&iPin=NCFC0005&SingleRecord=True
(2011) Bacillus Cereus. Absolute Astronomy. Retrieved March 2, 2011, from
http://www.absoluteastronomy.com/topics/Bacillus_cereus
(2002, April 21). Infectious Disease; U.S. Army to test company's antibiotics as potential antibioterrorism drugs. Medical Letter on the CDC & FDA, 8, Retrieved march 2, 2011, from
http://elibrary.bigchalk.com