Identifying Bacteria Through Staining Methods.
Connor Davis
Department of Biology, Francis Marion University, Florence SC
Abstract
In the study conducted, I collected, cultured and identified bacteria through gram staining
techniques. Based on the results from the biochemical test, I discovered the bacteria was
Staphylococcus.
Introduction
Microbiologist can identify bacteria in a number of ways; however, I focused on gram staining
which was invented by Hans Christian Gram in 18841 This allows us to distinguish between
gram negative and gram-positive bacteria which can help us identify our bacteria for the purpose
of this experience.
Methods
In the experiment I poured melted nutrient agar into a petri dish and allowed it cool off, I
swabbed the screen of my iPad and streaked it across the petri plate. I then placed the plate
upside down and incubated it at 37 degrees Celsius.
After the incubation I had selected one of the colony’s and described the anatomical morphology
of it. I then used a sterile transfer loop which was sterilized using dry heat, and transferred a
colony of bacteria to an agar slant and incubated that. With the rest of the colonies of the bacteria
I decided to gram stain them.
To use this method, I dried and heat fixed the bacteria on a slide then applied crystal violet for 30
seconds, rinsed the slide off with water, applied iodine for 30 seconds. I then used ethanol for
around 5 seconds and then rinsed with water. Finally applying safranin for 30 seconds and
rinsing and drying the slide. The slide was then examined at 100X under the microscope and I
described the stain results and that of the cells morphology.
After the agar slant finished its incubation, I used a sample of the bacteria for the use of several
other biochemical test to more accurately describe the bacteria using its biological morphology.
Using a sterilized transfer loop, I inoculated glucose, sucrose, and lactose fermentation tubes
with the bacteria. I also streaked a line of bacteria on a starch agar petri plate and then incubated
that and all of the test tubes and 37 degrees Celsius.
After the incubation, I had interpreted the results from the bacteria test2. I recorded any changes
in color and anything else that didn’t seem natural to the test tubes from the beginning of the
incubation. I also had poured iodine on the starch agar petri plate and interpreted results from
that, and finally adding a small amount of hydrogen peroxide to one sample of bacteria to test for
a catalase reaction.
Results
The results for the test from identifying bacteria are provided in the tables 1,2,3 listed below,
based on the results collected from the bacteria from the iPad they were staphylococcus. 3
Table 1. Unknown bacteria colony morphology
Color White creamy
Size (mm) 10mm
Form Irregular
Margin Undulate
Table 2. Unknown bacteria cell morphology
Bacteria Source Stain Reaction Cell Morphology
iPad Gram positive Staphylococci (clustered)
Table 3. Biochemical Properties of the undiscover bacteria
BIOCHEMICAL TEST RESULTS/ DESCRIPTIONS
Glucose Fermentation Acid produced/ no gas
Sucrose Fermentation No Acid/ no gas
Lactose Fermentation Yellow/ acidic
Litmus Milk Reactions Skipped
Protein (Gelatin) Hydrolysis Liquid positive for digestion
Starch Hydrolysis Yellow positive for digestion
Catalase Positive
Discussion
Staphylococcus is a leading cause of bacterial infections worldwide within the medical systems. 3
It can cause longer hospitalization depending on the patients’ medical history and can cause
diseases like staph infection. It tends to live on the skin and body however usually harmless can
enter the body through a cut making it the leading cause of skin infections. 4
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and satellite laboratories that do not have microbiology. Clin Chem Lab Med. 2017 Mar
1;55(3):458-461. doi: 10.1515/cclm-2016-0556. PMID: 27658158.
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hypervirulent clone. J Med Microbiol. 2015 Dec;64(12):1462-1473. doi:
10.1099/jmm.0.000185. Epub 2015 Oct 6. PMID: 26445995.
“Staphylococcus Aureus - Golden Staph.” Better Health Channel, Department of Health &
Human Services, 14 Sept. 2000,
www.betterhealth.vic.gov.au/health/conditionsandtreatments/staphylococcus-aureus-
golden-staph.
