Identification and Characterization of Unknown Bacteria

Christy Wise
BIOL 215
March 11, 2016

I.

Abstract
In this experiment, biochemical and morphology tests were performed on
unknown bacteria so that its characteristics could be matched up with a genus. After
gathering the results from the tests performed, it was determined that the unknown
bacteria was Staphylococcus. Determining the species of microbiota is especially
important in a healthcare setting so that the proper medication can be prescribed as

II.

treatment.
Introduction
There are many species of bacteria commonly found on the body, and this
experiment was performed to identify the specific genus and species of a sample that
was isolated from the ear lobe. A variety of lab and diagnostic tests were performed to
determine the bacterias traits and then to later use bacteriology charts to assign a
species. Virtually every part of the human body is home to bacteria, and most are
beneficial or harmless. In fact, in a study done in Norway, it was found that only 5%
of males and 15% of females did not have any microbial flora on their outer ear canal
(2). In a healthy ear, it is common to find microbe species such as Staphylococcus
epidermis, Staphylococcus aureus, and Streptococcus saprophyticum. The most
prevalent microbe found on the outer ear is Candida albicans, a type of fungus (3).

III.

Results and Interpretation

The first tests performed were the bacterial sample collection and isolation.
These tests were used to isolate a sample of unknown bacteria from the body with a
sterile swab so that no bacterial contamination occurred. After collection of the
bacterial sample, it was swabbed onto the petri plate and incubated to allow the
bacterial colonies to form. After incubating, there were visible colonies on the plate

that could be used to carry out the experiment. The growth of colonies was evidence
that bacteria was present on the outer ear lobe.
Next, the colony morphology was observed from the incubated bacterial
sample. The purpose of observing the colonys morphology was so that the bacteria
could be characterized based on its color, size, shape, and texture. The colonies from
the bacterial colony tested were bright yellow in color with a 5 mm diameter. They
had a circular form and entire margin. The test results from the colony morphology
were used to identify the type of bacteria present.
A gram stain was then performed on the unknown bacteria to determine
whether the bacterial cell was gram positive or gram negative. After staining the
bacteria and observing the sample using a microscope, it was determined that the
bacteria was gram positive because it was stained a purple color. The results of the
test concluded that the bacterial cell contained peptidoglycan in its cell was. This
characteristic is helpful for differentiating between different species of bacteria.
The cell morphology of the bacterial sample was observed using a
microscope. The cell morphology describes both the shape and formation of the
colonies and is useful for contrasting different species of bacteria. The unknown
bacteria tested was found to be staphylococci, meaning that the colonies were circular
shaped and in a clustered formation.
Finally, biochemical tests were performed on the unknown bacteria sample.
Many species of bacteria are able to metabolize carbohydrates such as sucrose,
lactose, and glucose to fulfill their energy requirements (1). The first set of
biochemical tests performed were Fermentation Tests, which tested if the bacteria was
able to ferment each type of carbohydrate. Whether or not the bacteria produced acid
(positive = yellow, negative = red) or gas (positive = bubble, negative = no bubble)

determined the results. For the glucose fermentation test, the bacteria tested positive
for acid production and negative for gas production. For sucrose fermentation, it
tested positive for acid production and negative for gas production. For lactose
fermentation, the bacteria tested negative for both acid and gas production.
The next biochemical test performed was the litmus milk test, which
determines whether lactose is fermented and if Casein protein is coagulated. Litmus
milk has several parts involved in testing: lactose (milk sugar); casein (milk protein);
and litmus (a pH indicator that is purple to blue at neutral to alkaline pH and pink
under acid conditions) (4). The unknown bacteria tested positive for protein
coagulation because curds were formed.
The nutrient gelatin test was performed on the unknown bacteria to determine
the bacterias ability to digest protein. After inoculating the gelatin tube with
unknown bacteria, the gelatin was liquefied. The results show that the unknown
bacteria was able to digest protein.
The final biochemical test performed was the catalase test, which was done by
dropping hydrogen peroxide on a sample of the unknown bacteria. The catalase test
determines if the bacteria reacts with hydrogen peroxide and produces the enzyme
catalase (1). The unknown bacteria produced bubbles when it came in contact with
IV.

hydrogen peroxide, meaning that it reacts and produces catalase.

Identification and Real World Application
After utilizing the results from the tests performed on the unknown
bacteria, it was determined that the genus of the unknown bacteria was most
likely Staphylococcus. Staphylococcus appears in pairs, singly, or in clusters and
may be white, yellow, or orange (5). The unknown bacteria were clustered and
yellow in color, and also matched all of the biochemical test results for
Staphylococcus according to Bergeys Manual. Another possible result could be

Micrococcus because the unknown bacteria matches up with all of the

biochemical test results and cell morphology. The only distinguishing factor
between the two genuses is that Micrococcus tends to be more orange or red in
color. Although the unknown bacteria colonies were a bright yellow color, it is
still possible that the unknown bacteria could be Micrococcus.
Staphylococcus is a genus to the well-known pathogens, Staphylococcus
aureus and Staphylococcus epidermis. Staphylococcus aureus is a type of
nosocomial infection that is usually acquired in the hospital through catheters,
breathing tubes, and IVs. Because it is able to form a biofilm, Staphylococcus
aureus is very difficult to prevent and treat (6). Staphylococcus epidermis is an
opportunistic nosocomial pathogen that usually only affects those with
compromised immune systems and indwelling medical devices (7). It causes
nosocomial infections almost as commonly as Staphylococcus aureus.
It is extremely important in a healthcare setting to be able to distinguish
between different illnesses and the microbiota that cause them. For example, in
order to determine the type of medication a patient should be prescribed, you
would need to know what type of microbiota is the underlying cause. A patient
with a fungal infection would need an antifungal medication, while a patient with
a bacterial infection would need to be prescribed an antibiotic. As a nurse, these
tests would be used to diagnose a patient so that the proper medication could be
prescribed. For example, a patient who comes to the clinic complaining of
vomiting and bloody stool would need to be tested for bacteria such as E. coli.
The tests used to determine the genus of the unknown bacteria would be too slow
to use in the clinic because most took at least 24 hours. Some advanced methods

of testing for microbiota in a clinical setting are the Enterotube system and rapid
Malaria test. Most rapid clinical tests take between a few minutes to an hour to
reach a diagnosis. Speed of diagnosis is crucial in a clinical setting because it is
important to be able to prescribe the patient a medication quickly.

Works Cited
(1) Pryor, G.S. and McCumber, L.J. 2008. Microbiology Laboratory: Fundamental Skills
and Experiments. RLSimonson Studios, Kearney, NB.
(2) "Result Filters." National Center for Biotechnology Information. U.S. National Library of
Medicine, n.d. Web. 10 Mar. 2016. <http://www.ncbi.nlm.nih.gov/pubmed/2216049>.
(3) "Microbes of the Ear." Microbe Wiki. Kenyon.edu, n.d. Web.
<https://microbewiki.kenyon.edu/index.php/Ear>.
(4) "LitmusMilk." LitmusMilk. N.p., n.d. Web. 10 Mar. 2016.
<http://web.clark.edu/tkibota/240/Unknowns/LitmusMilk.htm>.
(5) Bergey, D. H., and John G. Holt. Bergey's Manual of Determinative Bacteriology.
Baltimore: Williams & Wilkins, 1994. Print.
(6) "Result Filters." National Center for Biotechnology Information. U.S. National Library of
Medicine, n.d. Web. 10 Mar. 2016. <http://www.ncbi.nlm.nih.gov/pubmed/9597249>.
(7) Otto, Michael. "Staphylococcus Epidermidis the accidental pathogen."Nature
Reviews. Microbiology. U.S. National Library of Medicine, n.d. Web. 10 Mar. 2016.
<http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2807625/>.