Ridhima Phukan

FDSC 450
Dr. Dirks
S. aureus Lab
PURPOSE:

Staphylococcus aureus is part of the natural flora of humans. It is a gram-positive

facultative aerobe. It grows best at room temperature and can grow in high salt. Thorough
cooking destroys the bacteria itself but the toxin S. aureus produces is extremely resistant to heat,
refrigeration, and freezing, and the toxin is what makes humans sick. It gets into our food from
food handlers, especially from deli salads. Thus, we chose our sample to test to be a deli chicken
salad.
METHODS: For our seemingly harmless lunchtime chicken salad, it was crucial to find the
best mediums to grow it on. Because S. aureus loves salt, we chose Mannitol Salt Agar, which
with a 7.5% salt content, it inhibits all non-halophiles. The other Agar was Baird Parker Agar
which has selective agents like glycine, lithium chloride, and sodium tellurite to grow S. aureus
but not other unwanted bacteria.
We weighed 10g of the deli salad and added 90mL of saline, stomached for two minutes
and diluted up to -4, and incubated the plates at 37C for 24h. The next step was testing for
coagulase because its a virulence factor for S. aureus. We added 0.2mL of each culture to a
0.5mL coagulase plasma with EDTA, incubated at 37C for 4h.
RESULTS:

With Mannitol Salt Agar, mannitol is used by S. aureus as a carbon source, and

the phenol red is a pH indicator. If the organism can ferment mannitol, it will turn yellow. Only
our -1 dilution had growth, with 30 cream-colored colonies. None of them turned yellow and so

we estimated our detection limit in CFU/g by assuming there was 1 colony on the lowest
dilution.
For the coagulase test, we used another groups result whose specimen tested positive and
clotted because ours did not.
DISCUSSION:

Its unfortunate for learning purposes that we didnt test positive for

coagulase, but its reassuring for my laziness and habit of buying store-made salads. As with
many attempts in our adventures in food microbiology this quarter, we cant ascertain exactly
why this happened but we were able to see positive results from other groups.