Prelims Myco

MYCO 311 SUBCUTANEOUS MYCOSES
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OUR LADY OF FATIMA UNIVERSITY 3 REFERENCE MATERIALS: PPTS: JOSUE L. DELA CRUZ, RMT, MLS (ASCPi) Cm COLLEGE OF MEDICAL LABORATORY
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Mycetoma
01
SUBCUTANEOUS MYCOSES
 mycetoma is a chronic granulomatous
Mycetoma
infection that usually involves the
 Pseudallescheria boydii, ______________ but may occur in any part
 Acremonium spp. Exophiala jeanselmei, of the body. infection gradually
Curvularia spp., progresses to involve the bone, muscle,
 Cladophialophora bantiana,Madurella or other contiguous tissue and ultimately
spp., requires amputation in most progressive
 Trematosphaeria grisea cases.
(Previously Madurella grisea.)  tropical and subtropical regions of the
world whose out door occupations and
Chromoblastomycosis- failure to wear protective clothing
predispose them to trauma
 Cladophialophora spp.,
 Phialophora spp., Fonsecaea spp. Etiologic agents of eumyotic mycetoma
Phaeohyphomycotic  Pseudallescheria boydii and

Acremonium spp.,-causative agents of
 Alternaria spp., white grain mycetomas,
 Bipolaris spp.,  Exophiala jeanselmei, Curvularia spp.,
 Cladophialophora bantiana,Curvularia Cladophialophora bantiana,
spp, Trematosphaeria grisea (previously
 Exophiala jeanselmei, Madurella grisea), and Madurella spp. -
 Exophiala dermatitidis, causative agents of black grain
 Exophiala spinifer, mycetomas
 Exserohilum spp.,
 Rhinocladiella mackenziei, Madurella mycetomatis
 Verruconis gallopava
(Previously Ochroconis gallopava)  is the most common fungal agent
associated with mycetoma.
Dematiaceous fungi  most common etiologic agent of white
grain mycetoma ____________________
 are characterized by dark coloration as
a result of their ability to produce Chromoblastomycosis
melanin and are known agents of
superficial and subcutaneous mycoses  chronic fungal infection acquired
 Humans and animals serve as accidental through traumatic inoculation of an
hosts after traumatic inoculation of the organism, primarily into the skin and
organism into cutaneous and subcutaneous tissue
subcutaneous tissues.  development of a papule at the site of
the traumatic insult that slowly enlarges
GROWTH RATE to form warty or tumorlike lesions
characterized as resembling cauliflower
 slow-growing dematiaceous molds capable of spreading through the
 rapid-growing dematiaceous molds lymphatic system
 lesions usually are confined to the feet
Two types of Mycetomas
and legs but may involve the head,
 Actinomycotic (bacterial) mycetomas face, neck, and other body surfaces.
( N AND S)
 Eumycotic (fungal) mycetomas can be
subcategorized as (WGEM AND BGEM)
T R A N S C R I B E D B Y: P R E C I O U S V H A N N M A N L A P A Z
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Common etiologic agents of diseases that may

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 Histologic examination of the lesion be caused by dematiaceous fungi
reveals characteristic _________________,
which are copper-colored, septate cells Mycetoma
that appear to be dividing by binary
Bacterial:
fission and resemble copper pennies
 These infections cause hyperplasia of the  Nocardia, Actinomadura, and
epidermal layer of the skin, which may Streptomyces spp.
be mistaken for _________________.
 Fungal brain abscess, known in the past White grain mycetoma:
as cerebral chromoblastomycosis, may
be caused by the dematiaceous fungi  P. boydii and Acremonium and Fusarium
 Fungi most often associated with spp.
chromoblastomycosis include
Black grain mycetoma:
 _______________,____________________,___
________________
 Madurella spp., Exophialajeanselmei,
and Curvularia spp.
Phaeohyphomycosis
Chromoblastomycosis:
 general term used to describe any
infection caused by a dematiaceous
 Cladophialophora, Phialophora, and
organism; it includes molds; brownish,
Fonsecaea spp.
yeastlike cells; pseudohyphae; and
hyphae. Phaeohyphomycosis:
 progressive soft tissue infection, brain
abscess, sinusitis, endocarditis, mycotic  E. jeanselmei; E. dermatitidis; and
keratitis, pulmonary infection, and Curvularia, Bipolaris, Alternaria, and
systemic infection Exserohilum spp.
 Symptoms often include headache,
neurologic manifestations, and seizures. Sinusitis:
The most common fungal isolates
 Alternaria, Bipolaris, Exserohilum, and
associated with neurologic
Curvularia spp.
manifestations
Mycotic keratitis and endophthalmitis:
Pathogenesis and Spectrum of Disease
 E. dermatitidis and Bipolaris and
 The spectrum of disease caused by the
Curvularia spp.
dematiaceous fungi ranges from
superficial infections (e.g., skin and hair) Brain abscess:
to emergent, rapidly progressive, and
often fatal disease (e.g., brain abscess)  C. bantiana, E. dermatitidis, and Bipolaris
spp
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 Mycetoma and Phaeohyphomycosis

 Direct examination of clinical specimens
from patients with a eumycotic
mycetoma or phaeohyphomycosis
demonstrates yellowish brown, septate
to________________.
 Observation of pigmented hyphae in
hematoxylin-eosin or unstained
histopathologic sections is presumptive
for a diagnosis of dematiaceous fungal
disease
 _____________stain used to detect fungal
elements in tissues stains fungi black,
which makes determining whether they
are hyaline septate or dematiaceous
septate molds impossible
Laboratory Diagnosis  Fontana-Masson stain, which stains the
melanin and melanin like pigments in the
Direct Detection Method cell walls of these organisms,
 Stains- dematiaceous fungal hyphae are Serologic Testing

seen in clinical specimens by direct
microscopic examination or by  Some serologic and skin tests may be
histopathologic examination of tissue useful for the diagnosis of allergy to
obtained during surgery or autopsy dematiaceous fungi. However, serology
 ___________________, 10% silver nitrate is not useful for the diagnosis of invasive
and ammonium hydroxide stains fungal dematiaceous fungal disease
elements brown to black in a red
Molecular Methods
background. This technique improves
the detection of melanin granules.  _______________________ not routinely
 Chromoblastomycosis- crapings from used for detection or identification of
crusted lesions added to 10% potassium these fungi, although these tests may be
hydroxide (KOH) available in research settings and
show________________(aggregation of specialized laboratories.
dark brown cells that resemble stones in  Nucleic acid–based sequencing of
a stone wall) ribosomal genes- used for the
identification of fungal isolates
Sclerotic bodies
 Matrix-assisted laser desorption
ionization time-of-flight mass
spectrometry (MALDI-TOF MS)-
successfully used to identify clinically
relevant fungal isolates including yeasts
and molds.
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colonies with a gray to olive to black

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White Grain Mycetoma. P. boydii and velvety or suedelike appearance.
 grows rapidly (5-10 days) on common

laboratory media. Initial growth begins as
a white, fluffy colony that changes in
several weeks to a brownish gray (the so-
called “mousy gray”) colony; the reverse
of the colony progresses from tan to dark
brown. Acremonium spp. that cause
mycetomas, such as Acremonium
falciforme, grow slowly and produce
gray to brown colonies.
Cultivation
MYCETOMA
 Black Grain Mycetoma. Colonies of
Madurella spp. and E. jeanselmei are
slow growing, unlike Curvularia spp.
 Colonies of Madurella spp. vary from
white (during the early phases of growth)
Phaeohyphomycosis
to olive brown; a brown diffusible
pigment is characteristic of this fungus.  Colonies of Alternaria spp. are rapidly
 Colonies of E. jeanselmei appear growing, fluffy, and gray to gray-brown
yeastlike and darkly pigmented (olive to or gray green.
black), but in time develop a more  Colonies Curvularia spp. produce rapidly
velvety appearance with the production growing colonies that resemble those of
of aerial hyphae. Alternaria spp.
 Colonies of Curvularia spp. produce a  Bipolaris spp. produce colonies that are
fluffy or downy, olive-gray to black gray-green to dark brown and slightly
colony, and growth is rapid. T. grisea powdery, as do Exserohilum spp.
forms slow growing, velvety colonies that  Cladophialophora bantiana produces
appear smooth or radially furrowed and long, poorly branched conidial chains.
dark gray or olive brown to black.  Rhinocladiella mackenziei produces
pale brown conidiophores with
elongated conidia on denticles
(projection or peg) and may produce
exophiala-like budding cells in culture
 Verruconis gallopava produces a
rustybrown to olive colony with one- to
three-septate condia on small denticles
Chromoblastomycosis
 Cladophialophora, Phialophora spp.,

and Fonsecaea spp., are all
dematiaceous These fungi are slow
growing and produce heaped-up,
slightly folded, darkly pigmented
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Antifungal Susceptibilities
01  Antifungal susceptibilities for melanized
fungi for most clinically relevant species
are known. However, interpretive
breakpoints have not been
standardized. Amphotericin B and the
azoles have demonstrated clinical
effectiveness against infections with
melanized fungi. Triazoles,
posaconazole, and voriconazole have a
broad spectrum of activity against most
of these fungi. Occasional treatment
failure of mycetoma has been
associated with the use of voriconazole.
 END. THANK YOU…
MYCO 311 OPPORTUNISTIC MYCOSES
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among those with HIV or AIDS in the

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OPPORTUNISTIC MYCOSES United States
JOSUE L. DELA CRUZ, RMT, MLS (ASCPi) Cm  The introduction of highly active
antiretroviral therapy (HAART) for
OPPORTUNISTIC ATYPICAL FUNGUS: patients with HIV has reduced the
incidence of disease.
 Pneumocystis jirovecii
 DNA testing demonstrate the detection
 Current Name:Pneumocystis jirovecii
of P. jirovecii in immunocompetent
 PreviousName:Pneumocystis carini
populations as well as additional groups
General Characteristics of patients with chronic underlying
disease.
 1999, the name of the organism that
causes a pneumonia in Pathogenesis and Spectrum of Disease
immunocompromised humans,
 After P. jirovecii is inhaled, the trophic
commonly called pneumocystis
form of the pathogen is believed to
pneumonia (PCP), was changed from
adhere to type I pneumocytes
Pneumocystis carinii to Pneumocystis
 The organisms replicate extracellularly
jirovecii.
while bathed in alveolar lining fluid
 P. jirovecii is an opportunistic, atypical
 the alveolar spaces fill with an
fungus that infects immunocompromised
eosinophilic foamy material, which can
hosts and mostly manifests as PCP.
be detected with hematoxylin and eosin
 Precise taxonomic categorization of P.
staining.
jirovecii has been challenging. P. jirovecii
 Described, this pneumonia was known as
originally was believed to be a
interstitial plasma cell pneumonia.
trypanosome
 Symptoms of PCP include a
 Several factors later supported the
nonproductive cough, low-grade fever,
notion that P. jirovecii was a protozoan
dyspnea, chest tightness, and night
parasite; its morphology is similar to that
sweats.
of protozoa, and clinically it responds to
 In patients without HIV infection, the
antiprotozoal drugs but not to antifungal
underlying conditions most commonly
drugs in patients with pneumocystosis.
seen as risk factors for this opportunistic
 P. jirovecii exists as three forms in its life
infection are asthma, chronic obstructive
cycle: the trophic form (trophozoite),
pulmonary disease (COPD), cystic
sporozoite (precyst), and ascus (cyst),
fibrosis, systemic lupus erythematosus
which is the diagnostic form.
(SLE), pregnancy, rheumatoid arthritis,
 Its cell membrane contains cholesterol
infection with Epstein-Barr virus,
rather than ergosterol
ulcerative colitis, and high-dose
 DNA sequences of P. jirovecii with those
corticosteroid therapy
of other fungi confirmed the placement
of P. jirovecii in the fungal kingdom, in Laboratory Diagnosis
the phylum Ascomycota.
Specimen Collection and Transport
Epidemiology
 Respiratory specimens from the deep
 P. jirovecii has a worldwide distribution portions of the lung, such as
and most commonly presents as bronchoalveolar lavage (BAL), are best
pneumonia in an immunocompromised for detection of P. jirovecii.
host. Pneumocystis is transmitted person-
to-person via airborne particles.
 Pneumocystis has been defined as the
most common opportunistic infection
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 The immunofluorescent method showed

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 A sputum specimen submitted for direct greater sensitivity than the other three
examination should be induced sputum but a smaller negative predictive value
obtained by a trained respiratory  Direct Detection of (1-3)-Beta-Dglucan
therapist; otherwise, the rate of false-  The ascus (cyst) cell wall component, (1-
negative results may be unacceptably 3)-beta-D-glucan, has been used to
high successfully diagnose infections with P.
 Additional specimens may be tracheal jirovecii.
aspirates, pleural fluid, transbronchial  Other fungi also secrete the molecule
biopsy, or cellular material from but in lower amounts.
bronchial brushings  There are several commercial assays
available; uses patient serum for the
detection of (1-3)-beta-D-glucan
 It is important to use additional
DIRECT DETECTION METHODS diagnostic information and confirmatory
STAINS
testing in conjunction with this test,
 The diagnosis of P. jirovecii pneumonia
because other yeast or fungi also
currently is based on the clinical
secrete (1-3)-beta-D-glucan during
presentation, radiographic studies, and
infection.
direct or pathologic examination of
respiratory samples or biopsy material Molecular Methods
 Giemsa stains the nuclei of all the various
life cycle stages as reddish purple with a  A variety of nucleic acid amplification
light blue cytoplasm. assays for P. jirovecii have been
 Cysts are more easily recognized than developed, including, most recently,
the trophic form and may be definitively real-time polymerase chain reaction
identified using a variety of stains such as (PCR) methods
calcofluor white, methenamine silver,
and immunofluorescent staining Cultivation
Four most common staining methods  P. jirovecii is very difficult to cultivate

outside the lung; therefore routine
1. Giemsa culture methods are not performed.
2. Immunofluorescent
3. Calcofluor white Serologic Testing
4. Methenamine silver
 Serology is not useful for the diagnosis of
pneumocystosis.
 Opportunistic Fungus: Yeast
GENERA AND SPECIES TO BE CONSIDERED
 Blastoschizomyces spp
 Candida spp.
 Cryptococcus spp.
 Pseudozyma spp.
 Rhodotorula spp.
 Saccharomyces cerevisiae
 Sporobolomyces spp.
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General Characteristics
Blastoschizomyces Spp.
 Yeasts are eukaryotic, unicellular
 Blastoschizomyces are white to cream-
organisms that are round to oval and
range in size from 2 to 60 mm. colored, moist colonies that may appear
 In general, the yeasts reproduce to have radiating margins.
 Blastoschizomyces is typically found in
asexually by blastoco_x0002_nidia
regions where the summers are hot and
formation (budding) (Figure 62-1) and
sexually by the production of ascospores dry with warm, wet winters. Infections
or basidiospores have been primarily identified in
 With certain environmental stimuli, yeast immunocompromised patients
can produce different morphologies. An
Candida spp.
outpouching of the cell wall that
becomes tubular and does not have a  Candida spp. are responsible for the
constriction at its base is called a germ most commonly encountered
tube opportunistic fungal infections.
 The yeast may produce a capsule  Candida infections are caused by a
resulting in a shiny or mucoid colonial variety of species. C. albicans is the most
appearance. commonly isolated yeast, but other,
 More recent diagnostic tools that have emerging species include Candida
been introduced for quicker glabrata, Candida parapsilosis, Candida
characterization of yeasts include tropicalis, and Candida krusei
CHROMagar for Candida  C. albicans was the most common yeast
isolated from infections, accounting for
at least 60% to 70% of yeast infections
Cryptococcus spp.
 C. neoformans-C. gattii complex has

been divided into the two species and
five serotypes
 Standard laboratory tests do not
differentiate among the serotypes of C.
Epidemiology neoformans and C. gattii.
 Canavanine-glycine-bromothymol blue
 Yeast are ubiquitous in the environment
(CCB) agar has been recommended for
and often are part of the microbiota of
the differentiation of serotypes A/D and
both humans and animals
B/C
 Immunocompromised patients are
 It is most associated with avian excreta,
particularly susceptible to infection from
particularly pigeons.
yeast that are colonizers of the normal
 C. neoformans is believed to be widely
endogenous human microbiome.
distributed in nature, and aerosolization
 Health care–associated infections can
is a prerequisite to most infections.
contribute to the increased
development of pathogenic yeast
infections as a result of person-toperson
transmission.
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Pathogenesis and Spectrum of Disease
Blastoschizomyces spp.
 Blastoschizomyces spp. is an emerging

pathogen that is often isolated from
patients with debilitating disease such as
leukemia, renal transplants, ambulatory
dialysis, and osteomyelitis.Isolation is
typically from the blood of
immunocompromised patients who are
neutropenic. The organism has also been
isolated in the normal microbiota of the
human skin and gastrointestinal tract.
Candida albicans
 Candidiasis is an infection caused by a
 Candidiasis is an infection caused by a Candida spp. It may include
Candida spp. It may include oroesophageal candidiasis, intertriginous
oroesophageal candidiasis, intertriginous candidiasis (in which skin folds are
candidiasis(in which skin folds are involved), paronychia,
involved), paronychia, onychomycosis,respiratory infections,
onychomycosis,respiratory infections, vulvovaginitis, thrush, pulmonary
vulvovaginitis, thrush, pulmonary infection, eye infection, endocarditis,
infection, eye infection, endocarditis, meningitis, fungemia or candidemia, or
meningitis, fungemia or candidemia, or disseminated infection
disseminated infection  Paronychia is an infection of the tissues
 Paronychia is an infection of the tissues surrounding the nails, and
surrounding the nails, and onychomycosis is an infection of the nail
onychomycosis is an infection of the nail and nail bed
and nail bed  Thrush, an infection of the mucous
 Thrush, an infection of the mucous membranes in the mouth, is considered
membranes in the mouth, is considered a localized infection.
a localized infection.
 Thrush can be seen in newborns, patients Non-albicans Candida
with human immunodeficiency virus
(HIV) infection, individuals with diabetes,  The other Candida spp. (also called non-
and patients undergoing chemotherapy. albicans Candida), once believed to not
cause disease, are emerging as agents
of infection in certain patient
populations.
 Candida glabrata - become resistant to
common antifungal drugs such as
fluconazole and echinocandins. C.
glabrata has been isolated from serious
infections including endocarditis,
meningitis, and disseminated disease
 Candida tropicalis has been shown to be
prevalent in patients with hematologic
malignancies, especially those who are
neutropenic.
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immunocompetent hosts—a major

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 Candida krusei-is inherently resistant to difference from C. neoformans, which
the azole class of antifungal drugs, causes disease in primarily
identification of this species is essential to immunodeficient hosts. Some speculate
proper clinical management of the that C. gattii is able to modulate the
patient. host’s immune system by reducing the
 Candida parapsilosis is the primary inflammatory response or evading the
cause of fungemia in the neonatal immune system completely
intensive care unit (NICU). C. parapsilosis
is also the second most commonly
isolated Candida spp. in positive blood
cultures.
Other non-candida spp.
1. Candida norvegensis
2. Candida pulcherrima
3. Candida rugosa
Pseudozyma spp.
4. Candida utilis
5. Candida viswanathii  Pseudozyma spp. are environmentally
6. Candida zeylanoides associated plant pathogens that appear
7. Candida lusitaniae as beige to tan, moist, wrinkled colonies
8. Candida catenulate on routine laboratory media
9. Candida ciferrii
10. Candida dubliniensis Rhodotorula spp.
11. Candida guilliermondii
12. Candida haemulonii  A normal resident microbiota of the
13. Candida kefyr human skin, Rhodotorula spp.
14. Candida krusei  resemble Cryptococcus spp., appearing
15. Candida lipolytica as round, oval-shaped, budding yeasts
16. Cryptococcus neoformans that produce capsules.
 The organism can typically be recovered
 Cryptococcosis is an acute, subacute, or from moist environmental sources such
chronic fungal infection that has several as shower and bathtub grout, shower
manifestations. curtains, and toothbrushes.
 Phenoloxidase, an enzyme found in C.  Three species are associated with cases
neoformans, is responsible for melanin of human infection, including
production. septicemia, meningitis, peritonitis, and
 C. neoformans infections an present peritoneal dialysis.
initially as a chronic or subacute
pulmonary infection. C. neoformans Saccharomyces cerevisiae
eventually makes its way to the central
 Saccharomyces cerevisiae is the
nervous system, where the yeast can
common yeast that is used in baking
cause cryptococcal meningitis.
and the preparation of a variety of food
Cryptococcus gattii products.
 The yeast has been linked to human-to-
 C. gattii was believed to be a variant of human transmission in association with
C. neoformans until genetic studies health foods and baking.
proved it to be a distinct species. Since Saccharomyces sp. has been isolated
the 1990s this organism has emerged in from cases of thrush, vulvovaginitis, and
the Pacific Northwest as a pathogen in BSIs
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secretions, can be valuable for making a

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01 diagnosis of cryptococcosis
Sporobolomyces spp.
 Sporobolomyces spp. appear as salmon

to pink on routine laboratory media.
 The yeast is ubiquitous throughout the
environment and has been isolated in
cases of skin, respiratory, eye, and
central nervous system infections.
 Three species, Sporobolomyces
holsatiscus, Sporobolomyces roseus, and
Sporobolomyces salmonicolor, have
been associated with human infections.
Laboratory Diagnosis
Antigen Detection
FOR CANDIDA
 CAD test for C. neoformans may be
 Direct microscopic examination of performed on cerebrospinal fluid or
clinical specimens containing Candida serum. In many laboratories this assay
organisms reveals budding yeast cells has replaced the use of India ink to
screen for C. neoformans.
FOR CRYPTOCOCCUS
Molecular Methods
 India ink preparation has been the most
widely used method for the rapid  Nucleic acid amplification tests (NAATs)
detection of C. neoformans in clinical have been developed for a variety of
specimens yeast species.
 This method delineates the large capsule  peptide nucleic acid–fluorescent in situ
of C. neoformans, because the ink hybridization (PNA FISH) kits for the
particles cannot penetrate the capsular detection of C. albicans, C. glabrata,
polysaccharide material. and C. tropicalis from positive blood
 Many laboratories have replaced it with cultures
the more sensitive cryptococcal latex  Real-time polymerase chain reaction
agglutination test that detects (PCR) methods are now commercially
cryptococcal antigen. available in the TaqMan system (Applied
 Microscopic examination of other Biosystems, Carlsbad, CA) and
clinical specimens, including respiratory LightCycler
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Cultivation 01
 Candida spp. produce smooth, creamy
white colonies, but some produce dry,
wrinkled, dull colonies. In 50% of autopsy-
proven cases of invasive candidiasis,
organisms could not be isolated from
blood culture bottles.
 C. neoformans is easily cultured on
routine fungal culture media without
cycloheximidE.The organism is inhibited
by the presence of cycloheximide at
25°C to 30°C.
 Colonies of C. neoformans usually
appear on culture media within 1 to 5
days. Serologic Testing
 The use of serologic techniques for the

detection of the cryptococcal
polysaccharide capsule
glucuronoxylomannan (GXM) may be
completed using latex agglutination or
enzyme-linked immunosorbent assay
(ELISA)
Rapid Urease Test
 The rapid urease test is a useful tool for

screening for ureaseproducing yeasts
recovered from
 strains of Rhodotorula spp., some
Germ Tube Test Candida spp., and Trichosporon spp.
hydrolyze urea over time, so a distinction
 The germ tube test is the most generally should be made between a traditional
accepted and economical method urease test, which takes an overnight
used in the clinical laboratory to identify incubation, and the rapid urease test.
yeasts
 Approximately 80% of the yeasts Rapid Trehalose Test
recovered from clinical specimens are C.
 The rapid trehalose test can be used for
albicans, and the germ tube test usually
presumptive identification of C. glabrata
provides sufficient identification of the
organism within 3 hours.
 Germ tubes appear as early hyphal-like
extensions that are produced without a
constriction at the point of origin from
the yeast cell
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Conventional Yeast Identification Methods
1. Cornmeal Agar Morphology

2. Carbohydrate Utilization
3. Phenoloxidase Detection Using Niger
4. Seed Agar
MYCO 311 SYSTEMIC MYCOSES
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 Outbreaks have occurred following

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SYSTEMIC MYCOSES contact with moist environments such as
streams and rivers and contact with
JOSUE L. DELA CRUZ, RMT, MLS (ASCPi) Cm decaying vegetation. Cases in the U.S.
occur most frequently in the Ohio and
 This fungal group is often acquired via Mississippi River basins. More cases occur
inhalation and can disseminate to any of in males than in females.
the body's organ systems.  B. dermatitidis can be cultured from
 Most systemic fungi are dimorphic, tissue or body fluids.
exhibiting a nonmould (e.g., yeast)
parasitic phase at 35-37°C and a mould Identifying characteristics
(or mycelial) saprobic phase at 25-30°C.
 The mould phase is characterized by the
Identifying Characteristics presence of single smooth walled, round
to oval conidia at the ends of short
 Identification is based on temperature conidiophores. The mould phase of B.
and medium requirements and colony dermatitidis can be confused with
and microscopic morphology. Scedosporium apiospermum or
 Most systemic dimorphic fungi are very Chrysosporium spp.
slow growers and require 3-7 to grow.
 Because the mould forms are highly  S. apiospermum is the causative agent of
infective, slants are used for culture. mycetoma and can infect the brain,
 Colonies are membranous and develop bones, eyes, lungs, etc.
tan aerial mycelia.  Chrysosporium is commonly considered
 Conidia identification is necessary in a contaminant.
species identification.  Yeast phase: Large, round, thick-walled,
 Conversion of dimorphic fungi from the budding yeasts with blastoconidia
mould to yeast phase is confirmation that
the fungus in question is dimorphic.
Culture
Systemic dimorphic fungi  At room temperature, initially a yeastlike

colony develops, and over time the
1. Blastomyces dermatitidis (blastomycosis) colony will become fluppy white to tan.
2. Coccidioides immitis  Conversion from the mould to yeast
(cocccidioidomycosis) phase requires 4-6 days.
3. Histoplasma capsulatum (histoplasmosis)
4. Paracoccidioides brasiliensis
(paracoccidioidomycosis)
5. Sporothrix schenkii
Blastomyces dermatitidis
 Blastomycosis is a respiratory infection

that can affect the skin and bones.
 Infections are acquired by inhalation of
conidia or hyphae and can be mild to
chronic.
 The precise environmental location of
this fungus is unknown.
PRELIMS TOPIC:
WEEK:
SCIENCE
Histoplasma capsulatum
CABANATUAN CAMPUS
01
Coccidioides immitis
 Histoplasmosis can be a fatal pulmonary
 Coccidioidomycosis (valley fever) is an infection but can also affect the spleen,
infection of the lungs, bones, joints, skin, liver, kidneys, bone marrow, and heart.
lymph nodes, central nervous system,  Infection is acquired by spore inhalation
and adrenal glands. from barns, chicken houses, and bat
 Infections can be acute or chronic and caves. H. capsulatum has been
self-limiting or requiring medications. associated with guano, in particular from
 Most infections in the U.S. are in the starlings and bats.
semiarid southwest desert region (Lower  Most infections occur in the southern and
Sonoran Life Zone). Infections are Midwestern U.S. and along the
sometimes called desert or valley fever in Appalachian Mountains. The major risk
the San Joaquin Valley of California, factor for infection is environmental
where many cases are diagnosed. exposure.
 Infections are often acquired through  invades the reticuloendothelial system.
spore inhalation from the environment.
Activities that increase airborne dust, Identifying Characteristics
such as plowing and construction, can
Microscopic appearance
facilitate transmission.
 The mould phase will show
Identifying characteristics
conidiophores at 90-degree angles to
o Microscopic appearance hyphae supporting smooth
macroconidia (8-16 (Jim in diameter)
 Branching thick-walled, rectangular
(barrel-shaped) arthroconidia with finlike edges (tuberculate).
 Tissue phase shows round, thick-walled Microconidia are small (2-5 (xm in
spherule filled with small endospores. The diameter) and round to teardrop
shaped
tissue phase can only be grown under
 Yeasts appear as small single-budding
special conditions in vitro.
cells that are unremarkable in
B.Culture morphology.
 In clinical specimens, yeasts are often
 At 37°C on SAB HI agar, colonies will found inside monocytes and
appear moist and white and turn fluffy macrophages.
white in about a week.
 As with all mould phase fungi, always use Culture
a biological safety cabinet to prevent
 On blood-containing media, the
inhalation of spores.
colonies are initially moist and develop
tan aerial mycelia.
 Mature colonies are woolly and velvety
and appear tan colored.
PRELIMS TOPIC:
WEEK:
SCIENCE
CABANATUAN CAMPUS
2) Culture
01  When grown on blood-containing media
at 35-37°C, the colonies are waxy,
wrinkled, and cream to tan colored.
 When grown on SDA or PDA at room
temperature, colonies are initially
smooth. Colonies become tan with aerial
mycelium.
Penicillium marneffei
 P. marneffei is unique among the

members of the genus Penicillium in that
it is dimoiphic and a true pathogen.
Identifying characteristics
Paracoccidioides brasiliensis  The yeast cells are are oval and small (3-
8 |xm) and resemble H. capsulatum.
 Paracoccidioidomycosis is a chronic
 At 22-30°C, structures typical of the
granulomatous disease of the lungs and
genus Penicillium develop.
skin that can spread to the liver and
 Green aerial mycelium and reddish-
spleen.
brown hyphae are produced along with
 Mostly found in South America
a red diffusible pigment.
 Acquired by spore inhalation or ingestion
Sporothrix schenkii
Identifying Characteristics
 Sporothrix schenckii has been shown to
Microscopic appearance
be a complex of numerous species.
 Yeast cells grown at 35-37°C are thick Those involved in human infection
walled, with multiple budding yeast cells include S. schenckii,
with very narrow necks.  Sporothrix brasiliensis, Sporothrix globosa,
 The mould phase exhibits mostly hyphae and Sporothrix luriei.
with intercalary and terminal  Sporothrix spp. have a worldwide
chlamydoconidia. distribution, and their natural habitat is
living or dead vegetation.
 Humans acquire the infection
(sporotrichosis) through trauma (thorns,
splinters, bites, or scratches), usually to
the hand, arm, or leg.
PRELIMS TOPIC:
WEEK:
SCIENCE
CABANATUAN CAMPUS
Cultivation
01
 The infection is an occupational hazard  The dimorphic fungi are regarded as
for farmers, nursery workers, gardeners, slow-growing organisms, requiring 7 to 21
florists, and miners; it is commonly known days for visible growth to appear at 25°C
as rose gardener’s disease to 30°C.
 However, exceptions to this rule occur
with some frequency.
 Occasionally cultures of Blastomyces
and H. capsulatum are recovered in as
short a time as 2 to 5 days when many
organisms are present in the clinical
specimen. In contrast, when a small
number of colonies of Blastomyces and
H. capsulatum are present, sometimes 21
to 30 days of incubation are required
before they are detected.
Laboratory Diagnosis  Coccidioides is consistently recovered
within 3 to 5 days of incubation, but
Antigen-Protein when many organisms are present,
colonies may be detected within 48
 Immunodiffusion methods (the hours
exoantigen test) may be used to identify
isolates of these organisms based on
precipitation bands of identity between
specific antibodies and fungal antigen
extracts.
 However, these assays have been
largely replaced by the more rapid
nucleic acid hybridization reactions.
Molecular Methods
 Nucleic acid amplification assays are not

routinely performed but are available in
some reference laboratories and in
research settings. Realtime or
homogeneous, rapid-cycle PCR assays
have been described for
 H. capsulatum, Blastomyces,
Paracoccidioides, and Coccidioides
species.
 These assays have proven suitable for
isolate identification.
 Reproducibility and specificity of these
assays must be thoroughly evaluated for
standardization before implementation
in clinical laboratories.

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MYCO 311 SUBCUTANEOUS MYCOSES

Phaeohyphomycotic  Pseudallescheria boydii and

Common etiologic agents of diseases that may

 Mycetoma and Phaeohyphomycosis

 Stains- dematiaceous fungal hyphae are Serologic Testing

colonies with a gray to olive to black

 grows rapidly (5-10 days) on common

 Cladophialophora, Phialophora spp.,

among those with HIV or AIDS in the

 The immunofluorescent method showed

Four most common staining methods  P. jirovecii is very difficult to cultivate

GENERA AND SPECIES TO BE CONSIDERED

 C. neoformans-C. gattii complex has

 Blastoschizomyces spp. is an emerging

immunocompetent hosts—a major

Other non-candida spp.

secretions, can be valuable for making a

 Sporobolomyces spp. appear as salmon

 The use of serologic techniques for the

Rapid Urease Test

 The rapid urease test is a useful tool for

Conventional Yeast Identification Methods

1. Cornmeal Agar Morphology

 Outbreaks have occurred following

Systemic dimorphic fungi  At room temperature, initially a yeastlike

 Blastomycosis is a respiratory infection

 P. marneffei is unique among the

 Nucleic acid amplification assays are not

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