Central Philippine University

College of Medical Laboratory Science

MLS 2309 Clinical Bacteriology

Activity No. 1- Preparation of Culture Media

LABORATORY DISCUSSION

1. The medical laboratory scientist was tasked to prepare a Blood Agar, in the middle of the
preparation he noticed that there was no Defibrinated sheep’s blood available, instead of using
defibrinated sheep’s blood he used human blood as a substitute. What will happen to the growth of
bacteria when the medical technologist decided to use the said agar? As a medical technologist
what are the considerations to follow when preparing a culture media?

Human blood, particularly expired citrated donor blood, should not be utilized because it may
include antibiotics and antibodies such as ASO or anti-M protein, which might impede the
development of S. pyogenes. Beta-hemolytic streptococci are inhibited by citrate. Infectious agents
can be found in human blood. Although S. pneumoniae, S. aureus, and S. pyogenes thrive on HuBA,
the colony sizes are modest, the morphology typically differs from that seen on animal blood agar,
and hemolysis is limited, according to the findings. Because colony size, colony shape, and hemolysis
are all important factors in identifying these organisms, there is a significantly higher risk that
organisms grown on HuBA may be missed or misdiagnosed, especially when other organisms are
present (e.g., for samples from the upper respiratory tract or skin swabs).

Your outcomes are influenced by how you treat your culture medium, from storing it before
preparation to raising the agar plate top to incubation. If the starting materials are of poor quality,
compromised, or polluted, selective media will not be able to detect infections. Reduced
growth/recovery rates, aberrant colonial morphology, inhibition of the target organism, inability to
suppress competing flora, and a shorter shelf life of the produced medium are all problems caused
by improper culture media preparation. A medical technologist must be well-trained and meticulous
since the preparation must be done correctly in order to facilitate microbial development. Individual
culture media components (powders, gels, and liquids) must be metered out properly according to
the culture media formulation recipe.

2. An amount of 50 grams is needed for 1000mL (1L) preparation of Mac Conkey Agar, How much agar
does a medical laboratory scientist need to weight if she wants to prepare a 630mL of distilled water?
Round off your answer to the nearest tenths place.

Enumerate the given values:

Show your solution

Values:
Weigth of media in “Directions”(g) = 50 g
Volume of distilled water in “Directions” (mL) = 1000 mL
Weigth of powder should we weigth (g) = x
Volume of media (distilled water) we need (mL) = 630 mL

Formula:

Weigth of media in “Directions”(g) Weigth of powder should we weigth (g)

=
Volume of distilled water in “Directions” (mL) Volume of media (distilled water) we need (mL)
 Central Philippine University
College of Medical Laboratory Science
MLS 2309 Clinical Bacteriology

Solution:

50 𝑔 𝑥
=
1000 𝑚𝐿 630 𝑚𝐿

630 × 50
𝑥 =
1000

31,500
𝑥 =
1000

𝑥 = 31.5 𝑔

References:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1594681/

https://pubmed.ncbi.nlm.nih.gov/16954271/

https://www.thermofisher.com/blog/food/five-tips-for-culture-media-preparation-success/

https://www.mt.com/at/de/home/applications/Laboratory_weighing/Culture-Media-Preparation.html

https://www.slideshare.net/HusseinAltameemi2/medical-microbiology-laboratory-culture-media-preparation
 Central Philippine University
College of Medical Laboratory Science
MLS 2309 Clinical Bacteriology