Villalon, Abdul Jaimejay BS Biology 1-3

ACTIVITY / ASSESSMENT

1) Illustrate the step-by-step method of preparing culture media.

2) Research the significance of the following:

a. Soaking the container in bleach.
Using bleach to disinfect surfaces is common these days, also considering the
pandemic. Bleach contains sodium hypochlorite as its active ingredient and according to
Northeastern University Environmental Health and Safety (2017), the concentration of
sodium hypochlorite found in the bleach is enough to kill bacteria, fungi, and viruses.
However, though it is proven to be true in most cases, the contact time and concentration
of bleach solution will have the final say if the surface is already
disinfected/decontaminated (DiGangi, 2015). The recommended concentration of water
and bleach is at the ratio of 1:10; i.e., 10 mL of bleach diluted with 100 mL of water. And
the surface is to be soaked in the solution for at least 10 minutes to be considered
disinfected/decontaminated. These practices were performed in the activity.
b. Keeping the container closed except when pouring the medium;
partially opening the container during pour-plating.
Keeping the container closed before pouring the medium is part of the aseptic
technique. It prevents the microbes from contaminating the container after its disinfection
keeping it sterile. Pouring the media into the container, though partially opened, will still
deprive the microbes of the necessary nutrients they need for them to thrive as opening
the container is near the flame, killing the nearby microbes suspended in the air (Sanders,
2012).
c. Use of potato in the experiment.
The potato in the experiment was used to supply nutrients for the microbes for
them to thrive and encourage sporulation. In actual laboratory experiments, Potato
Dextrose Agar (PDA) is used to inhibit bacterial growth for molds, fungi, and yeasts (Aryal,
2019). However, actual laboratory experiments are not feasible but home-based
experiments will suffice. The agar (gelatin) was only used in the experiment to solidify the
solution as a potato does have that ability.
d. Inverting the container after the media had solidified.
When the container is not inverted, water evaporation from the media will happen.
The vapor will condense on the lid of the container and will drop on the media surface.
This causes colony mix-up and will spread across the plate surface. This will result in
difficulty in counting and proper determination of microbial count. In addition, the
evaporation will promote dryness that can affect the microbial growth of the media. Also,
the lid, which is not inverted, might contain foreign microbes that are not intended to be
mixed with the media, this will cause an error in microbial count determination
(Choudhary, 2008).
References:
Aryal, S. (2019). Potato Dextrose Agar (PDA)- principle, uses composition, procedure,
and colony characteristics. Microbiology Info. Retrieved from:
https://microbiologyinfo.com/potato-dextrose-agar-pda-principle-uses-
composition-procedure-and-colony-characteristics/
Choudhary, A. (2008). Why Petri dishes inverted during incubation?. Pharmaceutical
Guidelines. Retrieved from:
https://www.pharmaguideline.com/2015/09/incubation-of-petri-dishes-in-inverted-
position.html
DiGangi, B. (2015). Guidelines for using bleach. Shelter Medicine. Retrieved from:
https://sheltermedicine.vetmed.ufl.edu/files/2011/10/Guidelines-for-Using-Bleach-
updated.pdf
Northeastern University Environmental Health and Safety. (2017). Retrieved from:
https://www.northeastern.edu/ehs/wp-content/uploads/2014/12/Bleach-Fact-
Sheet-Draft.ejc2_.pdf
Sander, E. (2012). Aseptic laboratory techniques: plating methods. US National Library
of Medicine National Institute of Health, (63). Doi: 10.3791/3064