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Unknown Lab Report
Unknown #3
Alisson Evans
April 27, 2023
Dr. Derek Gibbs
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Introduction:
The identification of certain bacterias is very important. The identification is important
because it helps you distinguish one organism from another and group similar organisms
together without contaminating or mixing bacterias to possibly create something even
more deadly. It is also important to be able to identify different bacteria because of the
fact that bacteria affects humans. Knowing which agents are causing different diseases
makes it easier to give treatment for certain bacterias. The purpose of this lab was to use
different testing reagents in order to identify the bacteria you were using from 4 different
bacteria. We used different methods in order to identify the bacteria that we were testing.
Procedures:
Streaking for isolation- . Sterilize the inoculating loop in the bunsen burner by clicking on the
loop and dragging it to the burner. Put the loop into the flame until it is red hot. Allow it to cool.
Pick an isolated colony from the agar plate culture and spread it over the first quadrant
(approximately 1/4 of the plate) using close parallel streaks. Flame the loop Turn the plate 90°
and lightly sweep the loop 1-2 times through the inoculated area, then streak into the next
quadrant without overlapping the previous streaks. Flame the loop. 6. Turn the plate 90°, overlap
the previous area 1-2 times, and streak into the next quadrant as in step 4. Flame the loop.Repeat
#6, streaking the remainder of the plate. Invert the plate and incubate at 37°C for 24 hr.
Citrate Test-
Obtain a slant of Simmons Citrate Agar. Using an inoculating needle, stab the organism
into the medium, then zigzag the needle back and forth up the surface of the slant.
Incubate the tube for at least 48 hours. After the incubation period, examine the slant.
Methyl Red Test- Prepare MRVP broth in test tubes. Inoculate the broth aseptically with 2 loopful
of respective bacterial culture. Label the test tubes with the name of the organism inoculated.
Incubate the test tubes for 48-72 hours. Add a few drops of methyl red indicator in the incubated
tubes. Observe the results
Gram Stain-
● Applying a primary stain (crystal violet).
● Adding a mordant (Gram's iodine).
● Rapid decolorization with ethanol, acetone or a mixture of both.
● Counterstaining with safranin.
Results:
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Test Type Purpose Reagents Results Picture
Citrate Test screens a bromothymol Negative
bacterial isolate blue is used
for the ability to which is green
utilize citrate as when acidic and
its carbon and blue when basic.
The conversion
energy source
of green to blue
is positive
because pH goes
up.
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Methyl Red used to glucose, Positive
Test determine which peptone,
fermentation dipotassium
pathway is used phosphate, and
to utilize dextrose.
glucose.
Gram Stain Used to Crystal Violet, Negative
determine Gram’s iodine,
the chemical Ethyl alcohol, &
makeup of the Safranin.
cell
wall.
Streaking for allow you to tell A source of
isolation if you are bacteria
working with a Inoculation loop.
pure culture A striker/lighter.
EMB Bunsen burner.
Agar plate
Paper towels.
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Flow Chart:
Identification Tests
Gram
Stain
Test
Voges-
Citrate Proskau Test Methyl
Test Red
Identification
: (Ecolli)
Conclusion:
In conclusion we found our bacteria to be E. coli. One of the main ways that we were able to
come to this conclusion is because of the negative gram stain with the rod shaped bacteria that
was not colonized. E. Coli is a bacteria that is commonly found in humans and animals. E coli
can cause severe stomach pain, vomiting, and diarrhea. One of the most common infections that
is caused by E coli bacteria is an urinary tract infection.
References:
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E. coli (Escherichia coli).” Center of Disease Control. 2016.
https://www.cdc.gov/ecoli/general/index.html
Tawde M, McLaughlin S. Implementing a Virtual Midterm to Identify Unknown Bacteria in a
Microbiology Lab Course. J Microbiol Biol Educ. 2021 Mar 31;22(1):22.1.96. doi:
10.1128/jmbe.v22i1.2579. Erratum in: J Microbiol Biol Educ. 2022 Sep 14;23(3): PMID:
34012500; PMCID: PMC8101437.
