Name: Marielle P.

Balo Date Performed: August 24, 2018

Section: 1 Date submitted: August 31, 2018
Group No.: 4

1. Why is it important to sterilize the inoculating instrument before and after each
inoculation?

It is important to sterilize the inoculating instrument before and after each inoculation to
progressively dilute the sample until individual well-isolated organisms are obtained. Also, the
loop is flamed to destroy any residual microorganisms (Dart, 1996).

2. List the possible sources of contamination that you should be concerned about while
transferring bacteria from one culture to another.

Contamination can be caused by airborne contaminants and the use of non-sterile equipment
(Ryan, 2002). Other common sources of impure culture are colonies picked from the selective
medium, which suppresses growth of most unwanted bacteria but does not kill them. (Colwell &
Grigorova, 1987).

3. Why is it necessary to flame the mouth of the tube before and after performing an
inoculation?

The external surface of the culture tube is also susceptible to contamination, and the interior part
of the tube may be contaminated when opened to perform inoculation. By passing the mouth of
the tube through a flame immediately after opening and before closing it with the lid, the risk of
contaminating the interior part of the tube together with the cultured organism is minimized
(Stainer, Adelberg, & Ingraham, 1977).

4. Differentiate the following terms:

a. Stock culture vs. Pure culture

A pure culture is defined as a population of organisms descended from a single cell and is
therefore separated from all other species. Once a pure culture has been obtained, it can be
maintained as a stock culture, a culture stored for use as an inoculum in later procedures (Nester,
et.al.,, 2004).

b. Aseptic vs. Sterile

Aseptic means to exclude contaminating microorganisms from an environment, while sterile is

an absolute term that pertains to the state of being completely free of all microorganisms and
viruses (Nester, et.al.,, 2004).
References
Colwell, R.R. and Grigorova, R.R. (1987). Methods in Microbiology: Current Methods for
Classification and Identification of Microorganisms (Vol. 19). Orlando, Florida:
Academic Press Limited.

Dart, R. (1996). Microbiology for the Analytical Chemist. Cambridge: The Royal Society of
Chemistry.

Nester, Anderson, Roberts, Jr., Pearsall, & Nester. (2004). Microbiology: A Human Perspective
(4th ed.). Washington: The McGraw-Hill Companies.

Ryan, J. (2002). Understanding and managing cell culture contamination. Acton, MA: Corning
Life Sciences.

Stanier, R., Adelberg, E., & Ingraham, J. (1977). General Microbiology. Old Woke, Surrey,
Great Britain: The McMillan Press Ltd.