4th Shifting Micro Lab Reviewer

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DIAGNOSTIC MYCOLOGY

Specimen Collection
A properly collected specimen is the single most
important step in the diagnosis of an infection.
1. Sterile collection method and devices
2. Quantity of material must be adequate, with
accurate and complete label.
3. Sample should be representative of the infectious
process.
4. Prompt transport to lab to avoid overgrowth of
fungal or bacterial contaminants.
5. Physician has suspected diagnosis for special
procedures or specimen treatment.
Specimen Collection
6. Collect the right specimen:
Mycoses Specimen

Superficial Skin and hair

Cutaneous Skin, hair and nails


Subcutaneous Biopsy, granules

Systemic Abscess, blood, CSF,


sputum
Specimen Collection
Skin •If scraping, scrape the area with active infection
•Adhesive tape may be used for areas with sporulating
fungi (for rapid mounting).
Nails •Ask patient to stop antifungals 3 days before collection.
•Scrape the inner layer of infected nail.

Hair •Scalp infection: Collect at the edges of the scalp lesion


•Select hairs that fluoresce, are broken or are scaly.

Recovery of bacteria and fungi is most significant if the agent is isolated from a
normally sterile site.
•Significant if (+): blood, CSF, joint fluid, pleural cavity
•Consider in the context of normal flora: respiratory tract, GIT, GUT, wounds
or skin
Laboratory Precautions
• Conidia/ spores remain dormant even when
aerosolized.
• Laboratory procedures must be done in an
biosafety cabinet (BSL 3)
• Remember: Inhalation is a common route of
infection.
Microscopic Identification
Procedure Remarks

Saline wet •Budding of yeast, hyphae, pseudohyphae


mount •Lack of contrast, difficulty in identifying
fungal elements

Treatment •Breaks down the tissue surrounding the


with 10% fungal mycelia to allow a better view of the
KOH hyphal forms
•Specimen can examined unstained
Microscopic Identification: Staining
Staining Procedure Remarks

Calcofluor white •Binds to cellulose and chitin in the cell walls of fungi
stain •Fluoresces under long-wavelength UV light
•May demonstrate morphology that is diagnostic of the species
•Ex. spherules with endospores in Coccidioides immitis infection
Periodic Acid Schiff •Stain tissue sections when fungal infection is suspected
(PAS) •Oxidizes hydroxyl in the cell wall carbohydrate
Gomori •Fungi appear black against a pale background
Methenamine •For screening of clinical speciments
Silver Stain (GMS) •Identify Pneumocystis jiroveci cysts
Giemsa, Wright’s •Detect intracellular Histoplasma capsulatum in blood smears, lymph nodes, lung, liver,
stain bone marrow
•Cell light to dark blue in color
Lactophenol cotton •Stain used to distinguish fungal growth and identify organisms by morphology
blue (LPCB) •Used in tease preparation
India ink, Nigrosin •Used to identify Cryptococcus neoformans capsule
Microscopic Identification

GMS staining, H. capsulatum PAS staining, H. capsulatum

Giemsa staining, H. capsulatum


Systemic mycoses

Coccidioidomycosis
Histoplasmosis
Blastomycosis
Paracoccidiomycosis
Systemic mycoses
• Caused by dimorphic fungi

• Primary systemic pathogens - cause infection in


both “normal” and immuno-compromised hosts

• Endemic pathogens- distrubution varies to their


mode of entry
Dissemination of fungus

Pulmonary
Inhalation DIssemination
infection

Grow as yeasts or spherules in vivo, as well as in vitro at


37oC; exist in a mold form in nature or in the laboratory at
25oC to 30oC and in a yeast or spherule form in tissues or
when grown or enriched medium in the laboratory at 37oC
Coccidiomycosis

Etiologic Agent:
Coccidioides immitis
Colonial morphology

Suede-like to downy, greyish white colony; Tan to brown in reverse


Microscopic Morphology

– Hyphae,barrel-shaped, arthroconidia
Clinical findings
Primary Infection:

- Asymptomatic in most
- Nodular lesions in lungs

Secondary (disseminated)
infection (1%):

- Chronic/fulminant
- Infection of lungs,meninges,
bones and skin
Diagnosis

Direct examination: KOH; H&E


Histoplasmosis
• Most prevalent pulmonary micotic infection in
humans and animals

• Etiologic agent:
– Histoplasma capsulatum

• Yeast cell in tissue (37oC)


• Hyphae, microconidia and macroconidia
(tuberculate chlamydosphore) at 25oC
Colonial morphology

Mukha siyang pangalagay ng make-up (?)


Microscopic Morphology (MOLD form)

• Large, rounded, single-celled, tuberculate


macroconidia and small microconidia of H.
capsulatum.
Microscopic Morphology (YEAST form)

• Numerous small narrow base budding yeast cells inside


macrophages
Clinical findings

Primary Cutaneous Infection


Diagnosis

DIRECT EXAMINATION (GIEMSA/ WRIGHT)


Diagnosis

• Tissue morphology of H. capsulatum var. capsulatum (left)


showing numerous small narrow base budding yeast cells
inside macrophages and H. capsulatum var. duboisii (right)
showing larger sized budding yeast cells
Blastomycosis
• Etiologic agent:
– Blastomyces dermatitidis

Yeast cell at 37oC

- Bud is attached to the parent cell by a broad base


(Blastomycosis= Broad base)

Hyphae and conidia at 25oC


Colonial morphology

Sabouraud’s Agar at room temperature


- White or brownish colony
Microscopic Morphology (YEAST form)

• KOH, H&E: YEAST CELLS


- Grocott’s methenamine silver method: clearly see
the yeast like cell’s
Microscopic Morphology (MOLD form)

• Branching hyphae bearing sprherical, ovoid, or piriform conidia


on slender terminal or lateral conidiophores
Clinical findings

Chest Radiograph with blastomycosis


( note for the reticulonodular infiltrate)
Clinical findings
Manifest as ulcerated
granulomas

- 70% of patients
- Haematogenous spread
- Painless
- Verrucous lesions with
irregular borders , or as
ulcers
- Most Frequent Sites: Face,
upper limbs, neck and scalp
Paracoccidiomycosis
• Etiologic agent:
– Paracoccidioides brasiliensis

At 37°C (in tissue): multiple budding yeast cells


(Ship-steering wheel appearance); the buds are
attached to the parent cell by a narrow base

At 25°C: hyphae and conidia


Colonial morphology

Sabouraud’s Agar at room temperature


- White or brownish colony
Microscopic Morphology (YEAST form)

Direct microscopic exam: KOH & H&E

• Multiple budding yeasts; the buds are attached to the


parent cell by a narrow base “steering wheels”
Clinical findings

Granulomas may become active leading to chronic,


progressive pulmonary disease or dissemination.

- Many patients present with painful oral sores


Clinical findings

• Ulcerated lesion
in pharyngeal
mucosa

• Extensive
destruction of
facial features
Penicilliosis marneffei

• P. marneffei: the only species of Penicillium that is a


pathogenic dimorphic fungus
• Mold phase in culture at 25°C: sporulating structures
that are typical of the genus
• At 37°C in culture and in tissue: P. marneffei grows as
yeast-like organism that divides by fission and exhibits a
transverse septum
• Yeast from is intracellular in vivo and, in this way,
resembles H. capsulatum, although it is somewhat more
pleomorphic and elongated and does not bud
Colonial morphology

Typical Penicillium morphology & diffusible red pigment is


highly suggestive.
Microscopic Morphology (YEAST form)

Elliptic fission yeasts inside phagocytes in buffy coat


preparations or smears of bone marrow, ulcerative skin
lesions, or lymph nodes is diagnostic
Clinical findings

Skin infections (Ulcerative skin lesions)


From net: Penicillium sp. Giemsa stain

Skin lesions in Penicilliosis


Superficial Mycoses

Pityriasis versicolor
Keratomycosis
Tinea nigra
Black piedra
White piedra
Superficial Mycoses
• Limited to the outmost layer of skin and hair
Pityriasis (Tinea) versicolor

Etiologic Agent:
Malassezia furfur complex
M. globosa
M. restricta
Pityriasis (Tinea) versicolor
Characteristic Lesion:
Discrete, serpentine, hyper or hypopigmented macules occurring on
the skin of the chest, upper back, arms and abdomen
Wood’s lamp observation of lesions
(+) = Yellow Fluorescence
(positive yang pictures ah haha)
M. furfur Morphology
“Spaghetti and Meatballs Appearance”
M. furfur Morphology
Broadly budding yeast cells
Bottle shaped with a collarette-like
thickening seen at the junction of the mother and
daughter cells
Short hyphae, yeast cells

Tight clusters of spherical


yeast cells (MEATBALLS)
admixed with hyphal
fragments (SPAGHETTI)
M. furfur Morphology

Tight clusters of spherical


yeast cells (MEATBALLS)
admixed with hyphal
fragments (SPAGHETTI)
M. furfur Morphology
Broadly budding yeast cells
Bottle shaped with a collarette-like thickening seen at the
junction of the mother and daughter cells
Short hyphae, yeast cells
10% KOH examination

Short unbranched hyphae and spherical cells are observed


10% KOH examination

Short unbranched hyphae and spherical cells are observed


M. furfur Culture Characteristics

Saboraud’s Dextrose Agar:


Yeast like colonies
Creamy consistency after 2- 4 days incubation at 35oC
Keratomycosis/Mycotic keratitis

Etiologic Agent:
Saprophytic fungi
Aspergillus
Fusarium
Alternaria
Candida
Histoplasma capsulatum
(+)corneal ulcers
Tinea Nigra

Etiologic Agent:
Exophiala werneckii (Hortaea
werneckii)
Tinea Nigra
Lesions appear as a
dark (brown to black)
discoloration, often
on the palm (d/t
melanin produced by
the fungi)

Brownish maculae
on palms, fingers,
face, soles of feet
E. werneckii Morphology
Branched, septate hyphae and budding yeast
cells with melaninized cell walls (brown in
color)
E. werneckii Morphology
Branched, septate hyphae and budding yeast
cells with melaninized cell walls (brown in
color)
E. werneckii Morphology
Branched, septate hyphae and budding yeast
cells with melaninized cell walls (brown in
color)
E. werneckii Morphology
Branched, septate hyphae and budding yeast
cells with melaninized cell walls (brown in
color)
E. werneckii Culture

Black colonies
on culture
medium
Black piedra

Etiologic Agent:
Piedraia hortae
Black piedra

Discrete, hard, dark brown to black


nodules on the hair shafts
Ectothrix hair infection
P. hortae Morphology

Septate pigmented hyphae, and asci


unicellular and fusiform ascospores
with polar filament(s)
P. hortae Morphology

Septate pigmented hyphae, and asci


unicellular and fusiform ascospores
with polar filament(s)

*Di ito yung totoo nyang


itsura, wala kasi akong
mahanap eh sorry guys. Pero
more or less, kamukha nya
yan. 
P. hortae Culture

Brown to black colonies


White piedra

Etiologic Agent:
Trichosporon beigelii
White piedra
Soft, white to yellowish nodules loosely
attached to the hair
Occur as a sleeve or collarette around the
hair shaft
T. beigelii Morphology
Intertwined septate hyphae
Blastoconidia and arthroconidia
T. beigelii Morphology
Intertwined septate hyphae
Blastoconidia and arthroconidia
T. beigelii Culture

Soft, creamy colonies


Cutaneous Mycoses

Dermatophytoses
Trichophyton spp.
Microsporum spp.
Epidermophyton spp.
Dermatophytes
In dealing with dermatophyte identification, we
need to consider two things: microconidia and
macroconidia. If you know the feature
associated with a specific dermatophyte
genus, G ka na bes! Since species within a
genus have almost the same features
Trichophyton spp.

Macroconidium: (few) Typically absent or present only in small numbers


Elongated and pencil shaped
Multi-celled, thin smooth walls
cylindrical/ clavate/ fusiform
In single/clusters

Microconidium: Elongated, club shaped or large balloon shaped forms


Admixed with the smaller oval or tear shaped microconidia
Trichophyton spp.
Trichophyton spp.

MACROCONIDIA HYPHAE MICROCONIDIA


Trichophyton spp.
Club-shaped
microconidia

Thin-walled
macroconidia

Teardrop
microconidia

MACROCONIDIA HYPHAE MICROCONIDIA


Trichophyton spp.
Identify nyo na yung
parts kaya nyo na yan
haha
Macroconidium:
T. rubrum
Uncommon, if (+), pencil-shaped
Thin, smooth walls

Microconidium: Tear shaped, regular in size


Distributed on either side of the hyphal
Strands ( bird on the fence appearance)
Macroconidium:
T. rubrum
Uncommon, if (+), pencil-shaped
Thin, smooth walls

Microconidium: Tear shaped, regular in size


Distributed on either side of the hyphal
Strands ( bird on the fence appearance)

“bird on fence” appearance


Sa T. rubrum mas evident yung few
macroconidia
Trichophyton spp. Culture

T. rubrum: Burgundy pigment

T. tonsurans: Flat, granular,


T. mentagrophytes: Fluffy, rugose colony with buff to tan
granular colony with less brown pigment
intense pigment than T.
rubrum
Microsporum spp.

Macroconidium: Fusiform
Present multinucleated with thick rough walls

Microconidium: Present in small numbers, unevenly dispersed, generally oval or


elliptical, no distinguishing morphologic features
Microsporum spp.
Microsporum spp.

MACROCONIDIA
HYPHAE
MICROCONIDIA
Microsporum spp.

Fusiform
macroconidia

In M. canis,
macroconidia may be
barrel-shaped

Thick-walled
macroconidia

Few oval
microconidia
MACROCONIDIA
HYPHAE
MICROCONIDIA
Microsporum spp.Culture

M. canis: Cotton/wooly colony with M. gypseum: Sugary, granular


lemon-yellow pigmentaround colony with cinnamon brown
growing periphery or underside of to buff pigment
colony
Epidermophyton spp.

Macroconidium: Clavate/Clubshaped
Present with 3-5 cells and thin smooth walls
Cluster in groups of 3 or 4
Chlamydoconidia typically present in older cultures
Colonies are usually flat and velvety with a tan to olive-green
tinge

Microconidium: ABSENT
Epidermophyton floccosum
Epidermophyton floccosum

MACROCONIDIA
HYPHAE
Epidermophyton floccosum
Thin smooth walled
macroconidia

Macroconidia clustered into


3 or 4

NO MACROCONIDIA
MICROCONIDIA
HYPHAE
E. Flocossum Culture

E. flocossum: Suede appearance colony with


gentle folds and
Khaki or green yellow pigments
Tinea (Ringworm/Dermatophytosis)

• Tinea barbae
• Tinea corporis
• Tinea capitis
• Tinea cruris
• Tinea pedis
• Tinea manuum
• Tinea unguium
Tinea barbae (beard)

Etiologic agent: T. mentagrophytes


Tinea corporis (body)

Has hemorrhagic borders


Etiologic agent: T. rubrum, T.
mentagrophytes, T. tonsurans
Tinea capitis (head)

Inflammatory ectothrix infection


T. mentagrophytes

Gray-patchringworm
M. audouini/canis
Black-dotringworm Fungatingexophytic masses(kerions)
T. tonsurans T. tonsurans,T. schoenleini, T.violaceum
Tinea cruris (groin)

E. floccossum
Tinea pedis (foot)

Mocassin

Vesicular

T. mentagrophytes
T. rubrum
Interdigital
E. flocossum
Tinea manuum (Hand)
Tinea unguium (Nails)

T. mentagrophytes
T. rubrum
E. flocossum

ONYCHOMYCOSIS: Aspergillus spp., C.


albicans, Geotricumspp
Subcutaneous mycoses
I. Sporotrichosis
II. Chromoblastomycosis
III. Mycetoma
IV. Phaeohypomycosis
Lymphocutaneous sporotrichosis

• Caused by Sporothrix schenckii


Sporothrix schenckii
• Thermally dimorphic
– Grows well on routine agar
media
• Young colonies
– Appear blackish and shiny
– Wrinkles and gets fuzzy with
age
• Strains vary in
pigmentation from
shades of black and gray to
whitish
Sporothrix schenckii (Mold)

• Hyphae: Narrow
hyaline septate
• Conidia: Abundant,
oval borne on delicate
• Or in rosette or daisy petal
conidophores
Sporothrix schenckii (Yeast)
• Spherica, oval or;
• Elongated (“Cigar-shaped”)
• With single or (rarely) multiple
buds
Sporothrix schenckii
• Asteroid body
– Often seen in tissue
– In H&E, consists of a
central basophilic yeast
cell
– Surrounded by
radiating extension of
eosinophilic material, w/c
are
depositions of Ag-Ab
complexes
and complement
Sporothrix schenckii
Asteroid body
• The spheric yeastlike cells
are surrounded by
Splendore-Hoeepli material
Chromoblastomycosis
• Caused by dematiaceous fungi
(fungi that have brown to black
melanin pigments in their cell
wall
• Wart-like
• Cauliflower-like
• With abscesses covering the
area
• With covering the warty
surface
“black dots”
Chromoblastomycosis
• Molds are morphologically diverse, and most
are capable of producing several different
forms when grown in culture.
1. Phialophora verrucosa
2. Cladosporium carrionii
3. Rhinocladiella aquaspersa
4. Fonsecaea pedrosoi
5. Fonsecaea compacta
Phialophora verrucosa
• Conidia produced
from
flask-shaped
phialides
with cup-shaped
collarettes
• Conidia (mature):
spherical to oval,
extruded from the
phialide and
accumulate
around it
Cladosporium carrionii
• Conidia: branching chains
by distal (acropetalous)
budding
• Elongated conidiophores
with long,branching chains
of oval conidia
• Species identified based on
differences in the length of
the chains and shape and
size of the conidia
Fonsecaea pedrosoi
• Polymorphic phialides
with
chains of blastoconidia
similar to Cladosporium
sp
• Sympodial,rhinocladiella
type of condition
• Most strains form short
branching chains of
blastoconidia as well as
sympodial conidia
Fonsecaea compacta
• Blastoconidia: almost
spherical, with broad
base connecting the
condiia
• Structures are smaller
and more compact than
those of F.pedrosoi
Rhinocladiella aquaspersa
• Conidia: lateral or
terminal, from a
lengthening
condigenous
cell (Sympodial
process)
– Elliptical to
clavate
shaped
Chromoblastomycosis
• In tissue, the fungi that cause
chromoblastomycosis ALL characteristically
form murifom cells (sclerotic bodies, Medlar
bodies) that are chestnut brown because of
the melanin in their walls
Medlar body
• Brown-pigmented murifom
cell of chromoblastomycosis
Mycetoma
• Atinomycetoma- caused by actinomycete
– More invasive
• Eumycetoma- caused by a fungus
– Pseudoallescheria boydii
– Madurella mycetomatis
Exophiala jeanselmei
• Dimorphic fungi with
brown or black color or
dry aspect
• Dark yeast cells, septate,
and branched hyphae
• Unicellular anneloconidias
accumulates themselves in
the apex of elongated annelids
and with terminal or intercalate
formation
Curvalaria geniculata
• Mycetoma granule

Hard,contain intertwined,
septate hyphae that are
typically distorted and
enlarged
at the periphery of the
granule
Curvularia geniculata
• Myetoma granule

hyphage may be
embedded in an
amorphous cement like
substance Splendore-
Hoeppli
materials which
interdigitates
at the periphery of the
granules
Phaeohypomycosis
• Term used to describe a
heterogeneous array of
fungal infections caused
by pigmented, or
dematiaceous, fungi
which are present in
tissue as irregular hyphae
rather than the sclerotic
muriform cells seen in
chromoblastomycosis
Phaeohypomycosis
Opportunistic Mycoses
• Includes:
o Endogenous opportunists – Candida (Candidiasis)
o Exogenous opportunists
Cryptococcosis
Aspergillosis
Mucormycosis
Pneumocystis pneumonia
Penicilliosis
Candidiasis
• members of the normal flora of the skin,
mucous membranes, and gastrointestinal
tract.
• most common systemic mycosis
• In tissue, (37 C)
o germ tubes
• In the environment (25 C)
o Blastoconidia, pseudohyphae, and
chlamydoconidia (chlamydospores)
Candidiasis

• Culture

On Sabouraud's dextrose agar colonies are On blood agar, after 24 hours of


white to cream colored, smooth, glabrous incubation , moist opaque colonies
and yeast-like in appearanc are seen with yeasty odor
Candidiasis

• Direct Microscopy: Mold form


Candidiasis
• Direct Microscopy: Yeast form
Candidiasis

Germ Tube Test


• rapid screening
test where the
production of
germ tubes by
the cells is
diagnostic for
Candida albicans
Candidiasis

From Murray:
CHROMAGAR
• Differentiation of Candida species
• The green colonies are C. albicans, the blue-gray
colonies are C. tropicalis, and the large, rough,
pale pink colony is C. krusei.
• The smooth, pink or mauve colonies are another
yeast species (only C. albicans, C. tropicalis, and
C. krusei can be reliably recognized on this
media; other species have colonies ranging from
white, to pink, to mauve
Candidiasis

Calcofluor white stain demonstrating budding Candida stained with Gomori


methenamine silver demonstrating
yeasts and pseudohyphae of Candida albicans budding yeasts and pseudohyphae
Candidiasis (Intertriginous Infections)
Candidiasis (Oral Thrush)
Candidiasis (Vulvovaginitis)
Candidiasis (Onchomycosis)
Candidiasis (Chronic Mucocutaneous
Candidiasis)
Cryptococcosis
• Caused by: Cryptococcus neoformans , Cryptococcus
gattii
• Both are environmental, basidiomycetous yeasts
• occurs in immunocompetent persons but more often
in patients with HIV/AIDS, hematogenous malignancies,
and other immunosuppressive conditions
• Possess large polysaccharide capsules
• C. neoformans – worldwide, isolated from dry pigeon
feces
• C. gattii – less common, associated with trees in
tropical areas
Cryptococcosis

• In culture,
Cryptococcus species
produce whitish
mucoid colonies
within 2–3 days.
Cryptococcosis

• Microscopically, in
culture or clinical
material, the spherical
budding yeast cells (5–10
μm in diameter) are
surrounded by a thick
nonstaining capsule
Cryptococcosis
Yeast cell
with thick
capsule
unstained by
India ink
• In tissue and upon staining
with India ink, the cells are
variable in size, spheric,
oval, or elliptic, and are From CDC: C. neoformans stained with India Ink
surrounded by optically
clear, smoothly contoured,
spheric zones or “halos”
that represent the
extracellular polysaccharide
capsule

From upper batch trans


From Murray
Aspergillosis

• Ubiquitous in nature
• A. fumigatus is most common human
pathogen
• Produces abundant small conidia that are
easily aerosolized
• In immunocompromised patients, the conidia
may germinate to produce hyphae that invade
the lungs and other tissues
• In the environment (25 C), long conidiophores
with terminal vesicles on which phialides
produce basipetal chains of conidia

From CDC
• Grow in culture as hyaline molds.
• On a gross level, the colonies of
Aspergillus may be black, brown, green,
yellow, white, or other colors, depending
upon the species and the growth
conditions (Murray)

• On Sabouraud's dextrose agar, colonies


are flat, spreading topography and
powdery to felt like texture
• It generally forms a blue green colony
with a white margin and is usually cream
on the reverse
Aspergillosis

• Grow rapidly producing


aerial hyphae that bear
characteristic conidial
strucures
• long conidiophores with
terminal vesicles on which
phialides produce
basipetal chains of conidia
From Murray From Jawetz
Aspergillosis

• Aspergilli grow as branched, septate hyphae that


produce conidial heads when exposed to air in culture
and in tissue.
• A conidial head consists of a conidiophore with a
terminal vesicle, on which are borne one or two layers
of phialides, or sterigmata
• Identification of indvidual species of Aspergillus
depends in part on the difference in their conidial
heads, including the arrangement and morphology of
the conidia (Murray).
Aspergillosis

• On direct examination of
sputum with KOH or
calcofluor white or in
histologic sections, the
hyphae of Aspergillus
species are hyaline,
septate, and uniform in
width (about 4 μm) and
branch dichotomously
(Jawetz)
Mucormycosis

• Mucormycosis (zygomycosis) is
an opportunistic mycosis
caused by a number of molds
classified in the order
Mucorales of the Phylum
Glomerulomycota and
Subphylum Mucoromycotina.
• These fungi grow rapidly on
laboratory media, producing
abundant cottony colonies.
Mucormycosis

• These fungi are ubiquitous


thermotolerant saprobes
(Jawetz)
• The leading pathogens among
this group are species of the Rhizopus sp.
genera
o Rhizopus
o Rhizomucor
o Lichtheimia
o Cunninghamela
o Mucor

Cunninghamela sp.
Mucormycosis

• The asexual spores of the


order Mucorales are
contained within a
sporangium and are
referred to as
sporangiospores.
• The sporangia are borne at
the tips of stalklike
sporangiophores that
terminate in a bulbous
swelling called the Rhizopus sp. showing sporangium and rhizoids (root
columella. like structure)
Mucormycosis

On Sabouraud's dextrose
agar colonies are colonies are
very fast growing, cottony to
fluffy, white to yellow,
becoming dark-grey, with the
development of sporangia.
Mucormycosis (Rhinocerebral infection)
• Direct examination or culture of
nasal discharge, tissue, or sputum
will reveal broad hyphae (10–15
μm) with uneven thickness,
irregular branching, and sparse
septations
Pneumocystis jirovicii
• Causes pneumonia in immunocompromised patients
• P. jirovicii is the human species
• P. carinii is found only in rats

CDC: Cysts in the smear of bronchoalveolar CDC: Histopathology showing Pneumocystis cysts
lavage material in the lung of a patient with AIDS
Pneumocystis jirovicii
• Thin-walled trophozoites and cysts,
which are thick-walled, spherical to
elliptical (4–6 μm), and contain four to
eight nuclei
• Cysts can be stained with silver stain,
toluidine blue, and calcofluor white
• Trophozoites and cysts are present in a
tight mass that probably reflects their
mode of growth in the host
• Contains a surface glycoprotein that can
be detected in sera of acutely ill or
normal individuals

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