Microbiology Lab

Thursday
Unknown Lab Report
Whitney Johnson
April 24, 2014




http://gal1.piclab.us/key/micrococcus


On April 10
th
a slide was prepared by washing, drying, and drawing a wax circle on the
middle. A smear of the unknown bacteria was added to the slide and then heat-fixed by
passing through the flame of a Bunsen burner. After the slide had air dried a gram-stain was
performed on the unknown bacteria sample using the following materials: crystal violet stain,
iodine, ethanol, & safranin. The heat fixed bacteria was flooded with crystal violet stain for one
minute and then rinsed thoroughly with distilled water. The slide was then covered with iodine
for one minute and rinsed with distilled water, after which ethanol was dripped onto the slide
for approximately 20 seconds until the liquid ran clear. It was immediately rinsed with distilled
water. Safranin was then added to the slide and allowed to set for 30 seconds, then rinsed with
distilled water. To remove the excess water, the slide was blotted with bibulous paper. A
microscope was set up to view the slide using oil immersion. Once the microscope had been
focused, a drop of oil was added to the smear. Using the high power lens on the microscope to
view the smear showed a spherical bacteria that retained a purple stain with minimal pink
around some edges. Due to the purple stain the bacteria was determined to be a positive
bacteria.
A catalase test was performed on the sample by using a sterile swab dipped in the
inoculated nutrient broth and adding three drops of H
2
O
2
to the swab. This caused the H
2
O
2
to be
broken down by the catalase enzyme produced by the bacteria causing bubbles on the end of the swab;
although this was difficult to see. Had this test resulted in no bubbles it would have shown that the
unknown bacteria was Enterococcus.
Following the instructions on the unknown flow chart an isolation streak was added to a
Mannitol Salt Agar plate. The MSA plate was put into the incubator to held for 24 hours for the
incubation period. On April 14
th
, the MSA plate showed no signs of bacterial growth and there was no
fermentation of mannitol. The plate would have turned yellow if mannitol fermentation had occurred
and would have indicated that a halophile such as Staphylococcus aureus was present. In conclusion,
due to the negative result of the test the unknown bacteria was determined to be Micrococcus.
Micrococcus is a gram positive bacteria that is spherical in shape and forms irregular clusters of
cells or pairs(. It is a strict aerobe and produces a catalase enzyme. This particular bacteria is intolerant
to salt and therefore will not grow on Mannitol Salt Agar. It also does not ferment mannitol. Colonies of
Micrococcus appear in varied shades of yellow and red.












References
Micrococcus. Shauna Russell. April 22 2014
http://filebox.vt.edu/users/chagedor/biol_4684/micrococcus.html

Microbiology Laboratory Manuel for BISI 2124. Kelli Hasting. Angie Reisch.
2013. (Lab 4 Page 1- Lab 8 Page 7)