Aseptic Technique Effect on Cultured Microbes

By
Samara Brown
Mikia Parris-Bean
&
Chavelle Dillon
Mrs. Alnisha Simmons
Biology 1122 Section 01
11 Oct 2016

Introduction
The first organisms to inhabit Earth were prokaryotes that lived 3.5 billion
years ago (Campbell, Reece, et. al. 2014). Due to natural selection these
prokaryotes continue to transform and diversify themselves. As a result of
this diversification people are exposed to many different types of microorganisms. Micro-organisms are basically microscopic organisms that can
only be seen with the help of a microscope. Micro-organism includes fungi,
viruses, protozoa, algae, bacteria, and many more. Although, in this lab we
focused more on the micro-organism bacteria. Since bacteria are more
widely distributed than any other group of organisms, they can be a friend
or foe. For instance, Our intestines are home to an estimated 500-1,000

species of bacteria . . . they vary in their ability to process different foods

(2014). But some bacteria cause About half of all human diseases (2014).
An example of bad bacteria would include Tuberculosis or Lyme disease. To
help identify harmful of helpful bacteria certain techniques were
implemented such as the Aseptic Technique or Gram Staining Technique.
The Gram Stain is used to classify bacteria based on the cell walls structure.
Gram positive bacteria have a vast amount of peptidoglycan due to simpler
cell walls that retain a purple dye. On the other hand Gram negative
bacteria have complex cell wall that have less peptidoglycan and does not
retain dye. Gram staining is a valuable tool in medicine for quickly
determining if a patient's infection is due to gram negative or to gram
positive bacteria (2014). The focus on the lab preformed was on Aseptic
Techniques which concentrated more on how to prevent contamination with
micro-organisms. The purpose of the experiment was to test the microbial
growth on different surfaces where microbes can form without
contaminating the micro-organism. We hypothesized that if we swabbed
the surface of a doorknob, lab counter, and table then the lab counter will
exhibit more microbial growth and colonies far greater than the door knob
and table.
Materials
Three Petri Dishes
Lab counter
Table counter
Swabs

Lab counter
Incubator
Marker

Tape
Gloves
Sterile Cotton

Methods
In order to begin the process of cultivating bacteria using the aseptic
technique, a pair of gloves was needed to cover the hands, not just for
protection but also to prevent contamination. Secondly sterile sealed cotton
was used to swab the surfaces of a table, a door knob and another sterile
cotton swab was used to swab the lab counter by dragging the cotton tip
over the surface. Thirdly, three separate petri dishes containing nutrient
agar were required. To prevent contamination from the environment the
dishes were slightly opened for a minimum amount of time. Each swab was
dragged over separate petri dishes. Both swabs were then removed from
each dish, placed in the seal from whence they came and was properly
disposed of. The dishes were closed and then sealed using a tape to cover
the edges of each petri dish. As soon as the lids were sealed, a marker was
used to label the bottom of each dish using the group initials, the date and
what was being swabbed. Finally, all three petri dishes were placed in the
incubator for a period of forty-eight hours.

Results
The illustrations below show how much bacterial growth was found on the
Lab counter, a regular table and a doorknob. After 48 hours in the
incubator, we checked the bacterial growth in the agar plates. Also, found
out how much colonies are there by percent. In Illustration 1 there were
95% of microbes on a lab counter; Illustration 2 showed 45% of microbial
growth on a table and Illustration 3 represented 25% of bacteria on a door
knob.

Illustration 1: The lab counters morphology formed an irregular shape. Its

elevation was noted as raised and the margin looked undulate. The coloration
throughout the whole agar plate was a light yellow.

Illustration 2: The tables morphology formed a filamentous shaped for one part of the agar plate
and a circular part on the other side. The elevation was flat and had a lobate margin. Similar to
the lab counter, the color was also a light yellow.

Illustration 3: The doorknobs morphology seems to have formed a rhizoid shape with flat elevation
and a curled margin. Its color was a pale yellowish color. Dot's of discoloration was noted.

Colonies of Surfaces Swabbed

0.45

0.95
0.25

Lab Counter

Door Knob

Table

Figure 1: Shows the amount of colonies of surfaces swabbed in percentages

Conclusion
By analyzing the results its clear to say that out hypothesis was correct in
stating that the lab counter will have more microbial growth then the table
or doorknob. In terms of the number of colonies our calculations were a bit
off. We predicted that a doorknob would have almost 75% of colonies
appear on the Agar plate. Factors that could have affected this prediction
were the people who touched the door. Some could have previously washed
their hands before touching the door knob, or some may have been wearing
gloves. However when working with the Agar plate for the doorknob, I
noticed some discoloration amongst 3 colonies. It could be assumed that
this was the result of contamination. That being said it may have influenced
some of the results. Based on the above results it can be concluded that the
main question what surface would grow more microbes and colonies is
answered. The Lab counter had more microbial growth then the table and
doorknob.
Literature Cited

Campbell, N. A., & Reece, J. B. (2014). Biology: A global approach. Pearson

Education Limited.