Skin Microbiome: Antiobiotic Resistance

Emily Cribas
Hypothesis. My bacteria will be more resistant to most of the antibiotics presented.
Prediction. If there is a small/no clearing area around each antibiotic disk, then
my bacteria is resistant to that particular antibiotic.
Treatment and Controls. P. acnes was prepared using an RCM culture. Cells
from this culture were mixed with top agar and poured onto 3 blood agar plates
to encourage growth. Each plate contained 4 antibiotic disks, except plate # 2
contained only 3 .
Plate #1

Plate #2

Plate #3

P - Penicillin
D - Doxycycline

C - Cloramphenicol
MI - Minocycline
Am - Ampicillin
SXT - Sulfamethoxazole
-Trimethoprim
TE - Tetracycline CF - Cephalothin
NA - Nalidixic acid
E - Erythromycin
frozen
CC - Clindamycin
Table 1. Antibiotic Disk Setup and Codes

Results. b

Figure 1. Plate #1
The first plate showed no zones of clearing, and no growth, but showed an area
absent of blood agar.
1

Figure 2. Plate #2
The second plate showed a strange growth with a big clearing, but otherwise,
no P. acnes bacteria grew on these plates at all.

Figure 3. Plate #3
The third plate showed bacterial growth around the NA and SXT plates.
Conclusions. For the first plate, no P. acnes bacterial growth was observed.
However, there was a single colony that grew between 2 disks that appeared to
be hemolytic, indicating a contaminant, most likely S. aureus, a common bacteria
found on the skin.
The second plate most likely contained the same contaminant in higher quantities because it was also hemolytic, with more colony growth around a majority of
the plate. No accurate results could be concluded from this plate.

The third plate showed non-hemolytic bacterial growth around the NA and
SXT, but showed no growth around MI and CC, indicating bacterial susceptibility
to these antibiotics.
Based on the results and observations, the following can be concluded:
My bacteria is resistant to NA and SXT. However, my bacteria is not resistant to
TE, E, P, and D. Inconclusive results were found for: C, CF, and Am. Overall,
my bacteria was less resistant than predicted, and was more susceptible to certain
antibiotics than not.
Key Question. One of the key questions the results from this experiment answers is the following:
You find two colonies growing in the zone of clearing around a disk of ampicillin. If you tested the resistance of these colonies to ampicillin, what would your
hypothesis be and why?
These two colonies are resistant to ampicillin because they still grow and proliferate with ampicillin present. If they were vulnerable to ampicillin, they could
not grow in an area where they are exposed to it, because it would interfere with
the bacterias ability to synthesize peptidoglycan, a vital layer needed for survival.