Mycology & Virology
What are Dermatophytes?  Three genera: Microsporum, Trichophyton
 Pathogen – fungi that require keratin for
growth.
 Is the name of a pathogen that causes an
infection of tissue that contain keratin
 These dermatophytes are also group of
molds that causes superficial mycoses of the hair,
skin and nails.
 Utilizes keratin for growth and nitrogen as
an energy source
 We will focus on human infectious diseases,
but they are able to cause infections in a range of
other animals as well.
 Dermatophytes can really spread in a
population.
o Can be found in hair, skin, nails
o MOT: direct contact or via fomites
 It can be either by direct contact
within the infected tissues such as
during physical interactions, as in
sporting events like wrestling, or
through objects that have come
into contact with infected tissues
such as nail clippers or hair brush.
o Risk Factors: age, participation in sports,
family history, contact with feral animals, etc.
 While anyone can get those types
of infections, as noted there are
certain risk factors (mentioned
above)
 These are certain behaviors such
as sharing equipment or personal
hygiene products that are not
properly disinfected can also lead
to these types of fungal
infections.
 In general, these are common
infections that affects significant
portion of the population and are
likely under diagnosed due to the
over-the counter remedies and the
lack of presentation to clinics.
Therefore it is known as having
self medication that they don’t
know the underlying possible
consequences of them doing that
in their treatment
 Fungi
o Eukaryotic organisms – cell wall,
true nucleus
 Undergoes cellular division
and Epidermophyton.
 They may produce large leaf or
club shaped sexual spores called
macroconidia
 They also produce small,
spherical asexual spores called
microconidia
 Both are from vegetative hyphae.
FUNGAL SPECIMEN COLLECTION,
HANDLING, AND TRANSPORT
 Appropriate specimens
o Primary criterion for accurate
diagnosis of mycotic or fungi
infections.
 Transported and processed without delay
o Many pathogenic fungi grown
slowly and delay in a processing
compromises specimen quality and
decreases the possibility of isolating
the causative agent as a result of
overgrowth by contaminants like
bacteria.
 SOP (Standard Operating Procedure) for
rejection of unsatisfactory and improperly labelled
specimen.
 Most common fungal specimens: Hair, skin,
nails, respiratory secretions, tissue, blood , bone
marrow (BM), & CSF.
o Almost any tissues or body fluids
can be submitted for fungal culture.
o The most common fungal specimens
which are hair, skin and nails, which
we will be using to identify the
dermatophytes.
LABORATORY DIAGNOSIS OF FUNGAL
INFECTIONS
 Direct microscopic examinations
 KOH Preparation
 KOH with Calcofluor White
 India ink
 Tissue stains
 Isolation methods – Culture
 Macroscopic examinations of culture
 Miscellaneous tests for ID of yeasts
 Other current methods used in the lab:
MALDITOF and other serological test for fungi
such as ELISA, LIA, Immunodiffusion and latex
agglutination tests.
 Molecular techniques: PCR and DNA
probes.
o Identification of fungi or fungi infections
has become more sensitive and specific.
Direct Microscopy
 Most common
Skin Scraping Specimen
 Specimen
o Skin lesions (skin scraping)
 Procedure:
o Clean the lesion and periphery with 70%
isopropyl alcohol prior to collection
because our goal is to decontaminate the
area from contaminants like bacteria
o Use a sterile scalpel or edge of a
microscope slide to scrape perpendicular
to the skin
o Scrape around the active edge where the
fungus is actively growing. The center of
lesion heals first so the lab results are
negative using this sample. If the lesion
is inflamed or with fissures, clean it with
sterile distilled water
o Collect skin scrapings in paper envelope
or petri dish or place between 2 slides
o Store at room temperature (RT)
o If patients are young children and are
scared of the scalpel, you can use scotch
tape to collect specimen for microscopy
and you may also collect most exudates
for Candida albicans
 Materials needed:
o 70% Alcohol
o Cotton Swab
o Scalpel
o Black cardboard/Petri dish
o Scotch tape
Outer Active edge (blue arrow)
 KOH wet mount
 Procedure:
o Purpose of KOH wet mount is to break
down the tissue, making it easier to view
fungal hyphae
o The remaining materials or sample is
inoculated directly onto the agar.
 KOH Wet Mount
o Potassium hydroxide (KOH)
o Used for the rapid detection of fungal
elements in clinical specimens
o 10-30% strong alkali
o Parker blue-black ink – fungal appear
bluish green
o Specimen, skin, hair, nails mixed w/
20% w/v KOH- softens, digests, and
clear the tissues
o Microscopic result: the presence of
hyphae and conidia (spore) of fungi
 Procedure:
o You can prepare a wet mount form a
specimen plus sterile water or NSS or
the specimen alone like the exudates, but
we can use KOH from 10-30%
depending upon the type of specimen.
For skin, we can use the lower
concentration (10%). For nails, you can
use a stronger concentration which is
30%
o KOH digests or dissolves protein, fats
and carbohydrates. The tissue clears and
makes the fungal cell wall. Which is
resistant to alkaline.
o KOH is a strong alkali. When specimens
such as skin, hair, nails or sputum is
mixed with 20%