DOMINGO, Jose Carlo R.
September 11, 2021
3CMT – 21
MT 6322 – MYCOLOGY & VIROLOGY LAB (OLA)
1ST Shifting Units (I-IV)
QUESTION 1
The mycology laboratory poses many risks to laboratory professionals because of the nature of the
pathogens being handled and the procedures and assays being performed on the said pathogens.
Identify at least 5 examples of risk control measures that can be applied in the mycology laboratory
and the rationale for its use. Tabulate your answers as shown below. The first row is given as an
example and must be deleted when submitting your answers.
Risk Control Measure Example
Administrative Controls Strict limitation of unauthorized
personnel into the laboratory
Substitution Bunsen burner replaced with
microincinerator when
sterilizing inoculating tool.
Engineering Controls Using biosafety cabinet or
laminar flow hood
Personal Protective Equipment Wearing gloves
Administrative Controls Having a strict schedule for the
maintenance, cleaning, and
inspecting of the equipment and
the laboratory.
Rationale
Ensuring that people who enter
the mycology laboratory are
knowledgeable of the
procedures and precautions to be
done in the laboratory
minimizes the risks.
Open flame sterilization poses
increased risk such as ignition of
combustible materials, personal
injuries, destruction of property,
and loss of life.
Having a BSC/LFH provides
personnel a safe work area that
maximizes their protection from
the microorganism, prevents the
contamination of the organism
they are isolating/culturing, and
protects the environment from
the organism.
Gloves are an essential PPE
when handling microorganisms
that can potentially infect
laboratory personnel. It creates a
physical barrier against the
organism and personnel to
prevent mycoses.
Regularly maintaining the
equipment and the laboratory
minimizes the risks from
exposure and equipment
malfunction.
QUESTION 2
There is currently limited availability of clinical mycology laboratories in the Philippines and their
utility is also low. Reflect on this and discuss what are the probable reasons on the limited
availability of facilities and tests related to clinical mycology in the country. What are the probable
actions that can be done to improve this current scenario?

It is evident that the Philippines lack infrastructure that is dedicated to clinical mycology as even
some HEIs share mycology sections within laboratories. One probable reason for this shortage
would be the relatively weak prevalence and low incidences of mycoses here in the Philippines, as
compared to infections by bacteria, viruses, and even parasites. Another reason could be of the
lack of interest in the field of mycology. As it is only a limited area of microbiology, it tends to be
disregarded and overshadowed by other fields in microbiology. That being said, actions should
still be done to improve the availability of clinical mycology laboratories. The government can
allot funds in building clinical mycology laboratories in areas that lack the section. Equipment and
resources can also be acquired to encourage students regarding clinical mycology. Lastly, creating
collaborative work between undergraduate students and graduate students can improve the interest
of the said people regarding clinical mycology, thus, can possibly lead to further construction of
clinical mycology laboratories.

QUESTION 3
Various samples can be submitted for mycological testing from patients. Provide detailed notes
that are essential to remember for the collection of the following samples:
Sample Notes Regarding Sample Collection
Skin scrapings • Should be collected at the rim (outer edge) of
surface lesions because most healthy hyphae are
located there
• After collecting a sample, it should be put in
between to glass slides and then wrapped in paper
or put in an envelope for transport.
CSF • Should be concentrated by centrifugation before
inoculation
• >5ml of sample, then it may be filtered through a
membrane filter
• Should not be put in medium that has antimicrobial
agent as it is generally sterile
Urine • Should be collected early in the morning
• Clean catch midstream urine collection should be
observed
 • Sediments after sample centrifugation is used to
inoculate in media and also in microscopic
examination via smear.
Blood • Lysing erythrocytes and leukocytes through
centrifugation can be done to look for molds and
yeast in sediment.
• Solid media and biphasic media can be used for
culture
• Blood-to-broth ratio is highly essential
Sputum • Should be collected in the morning (preferred)
• Should be contained in a sterile, screw-top
container
• Can be expectorated, but in cases where patients
have trouble, can be induced via nebulizers.
• Dacron swab is used if the sample is viscous
(mucolytic agent; N-acetyl-L-cysteine can be used)

QUESTION 4
Traditional approaches to laboratory diagnosis of medically-important fungi typically involve
microscopy and culture methods. However, evolving technology in the clinical laboratory paved
the way for more advanced methods such as automation and molecular based assays. Given 3
examples of these emerging technologies in mycology, their advantages and disadvantages
Mass spectrometry – Using mass spectrometry, samples can be examined in convenience in
regards to time and effort of the laboratory personnel. An example would be MALDI-TOF wherein
samples can be prepared easily and results are obtained readily. Disadvantages, however, include
the expensive cost of some machines, as well as, the strict requirement of a pure culture.
Whole Genome Sequencing – The advantages of genome sequencing include a very high
specificity, the capability of the procedure to identify organisms, and assessing drug resistance. It
also has a high tolerance to contaminated samples. However, the disadvantages of genome
sequencing pertain to its long turnaround time (TAT), costly procedures/equipment, the need of
high levels of expertise, and its complexity in general.
Hybridization – This principle uses fluorescently-labeled probes to anneal to its target and thus
producing fluorescence in its presence. An example would be the Fluorescence In-Situ
Hybridization (FISH) technique that has been used in different assays in clinical laboratories. The
disadvantage of this procedure would be the level of expertise required to execute t he procedure
due to its complexity.
REFERENCES:

Centers for Disease Control and Prevention. (2015, January 13). Hierarchy of controls. Centers
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GOVPH. Research Institute for Tropical Medicine. (n.d.). Retrieved September 11, 2021, from
https://ritm.gov.ph/reference-laboratories/national-reference-laboratories/mycology/.

Kozel, T. R., & Wickes, B. (2014, April 1). Fungal diagnostics. Cold Spring Harbor perspectives
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Mahon, C. R., & Lehman, D. C. (2019). Textbook of diagnostic microbiology. Amazon.

Retrieved September 11, 2021, from https://www.amazon.com/Textbook-Diagnostic-
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