Central Philippine University

College of Medical Laboratory Science

MLS 2309 Clinical Bacteriology

Activity No. 3
LABORATORY DISCUSSION

1. When inoculating samples onto an agar plate, what are the possible effects of the following missteps
on bacterial growth? Provide 1 example for each scenario.
a. Forgetting to flame the loop before getting a loopful of inoculums

The tube's medium may get polluted. While the test tube's cover is off, heating the mouth of
the tube generates an air current that helps prevent airborne germs from falling into the tube.
If bacteria from the air fall into the tube, the medium will be contaminated. If you use a fresh
or sterilized loop, it helps you to efficiently dilute the inoculum on the plate and generate
isolated colonies by spreading the inoculum thinner and more uniformly.

b. Using a loop while it’s still red hot when fishing samples

Aerosols may be produced when the heated loop comes into touch with the culture. When
the heated loop comes into contact with the liquid broth culture, part of the broth (and
bacteria) will boil quickly, resulting in a bacterium-laden aerosol. These germs in the air might
go into the student's lungs or settle on her skin. You haven't cooled the loop sufficiently if you
hear a hissing sound when you insert it into the broth culture. Excess heat has the potential to
destroy bacterial cells. To avoid harming the instruments, do not overheat them. When you
lay the inoculating loop on the bench top, be sure it doesn't burn the surface.

2. When placing the culture plates inside the incubator, you positioned the plates with their lids placed
on top of the agars. What is the possible consequence of this to bacterial growth on agar plates?

When you return the dish to the incubator, the medium evaporates, condenses, and forms
droplets on the cover due to the heat. If it falls on media again, the colonies may become mixed
up (spread out) and you will be unable to count them correctly. The droplets produced may
come into contact with pollutants on the dish's lid, resulting to a possible consequence of colony
contamination an improper microbial growth.
 Central Philippine University
College of Medical Laboratory Science
MLS 2309 Clinical Bacteriology

References:

https://blog.microbiologics.com/7-streak-plate-method-best-practices/

https://www.umsl.edu/microbes/files/pdfs/streakplates.pdf

https://www.pharmaguideline.com/2015/09/incubation-of-petri-dishes-in-inverted-position.html

https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/58
3859/Q_5i2.pdf

https://www.umsl.edu/microbes/files/pdfs/tipsforplates.pdf