Skin Microbiome: Host Specificty

Emily Cribas
Hypothesis. My bacteriophage will completely or almost completely infect other
bacteria.
Prediction. The phage titer from the original P. acnes strain will be the same
as those from other bacteria.
Treatment and Controls. The titer from the original bacterial sample is a positive control, because it my bacteriophage is known to completely infect this strain,
therefore, the titer count of the phage when exposed to this bacteria serves as an
accurate sample of the concentration of bacteriophage. The titer count was 1.2 x
109 pfu/ml.
6 plates were prepared with 6 different P. acnes strains gathered from other
groups samples. After their bacteria dried when mixed with top agar, serial dilutions of the bacteriophage (obtained using the skin microbiome bacteriophage
purification protocol) were done, and 5L of each diluted phage (up to 108 , were
pipetted onto each plate. After they dried, the plates were inverted and placed in
an anaerobic environment.

Figure 1. Example Group Bacteriophage

p df
Where p is the
vp
number of plaques counted, df is the highest dilution factor where plaques are
found, and vp is the volume of bacteriophage plated in mL.
Phage titer is calculated using the following equation:

Results. The first plate contained absolutely no phage growth. The ATCC 6919
sample contained more phage than the positive control. The 6th plate grew almost
as much as the positive control and less than ATCC. The other 3 grew less than
the control, and grew almost the same amount in each one.
1

Plate Phage Titer (pfu/ml)

1
2
3
4
5
6

0
6 107
1 107
1.8 107
8 109
1 109

Origin
Mary and Lexi
Raza and Noory
Katie and Ryan
Lu
ATCC 6919
Sally and Lauren

Table 1. Phage Titer Per Bacterial Strain (Plate)

Conclusions. The ATCC 6919 is the most susceptible P. acnes strain out of all
of the ones tested, including my own isolated bacteria (the positive control). Mary
and Lexis sample were completely resistant to my bacteriophage, while Sally and
Laurens sample were almost completely susceptible to it. The other titer counts
were not as high as predicted. My bacteriophage is not as resistant towards any
of these strains as the positive control, but sample 6 was fairly close. Overall, my
bacteria infected almost all samples to varying degrees, indicating medium-high
specificity.
Key Question. One of the key questions the results from this experiment answers is the following:
Bacteriophage that infect P. acnes are being considered as a novel treatment
for acne. What properties would make a bacteriophage a good drug that could
treat acne in many different patients?
Bacteriophage target specific types of bacteria, in this case, P. acnes. This host
specificity characteristic of phage is a nice property of a drug, because it gets rid
of off-target effects, and is not dangerous in the sense that it can attack other
bacteria that may be essential to our microbiome and survival. A phage that is
effective in infecting many different strains of this bacteria would also be a good
characteristic to be able to market it to a variety of patients.