MYCOLOGY - VIROLOGY RTRMF.

MLS 3B
SUBCUTANEOUS MYCOSES forces and cation
exchange capacity
 Are chronic localized infections of the skin following
Sphagnum moss can
traumatic implantation of the fungal organisms into the
potentially harbour the
skin into the subcutaneous tissues.
chronic fungal
 The causative fungi are all soil saprophytes that has the disease sporotrichosis.
ability to adapt to the tissue environment and elicit/ cause
disease
Sporothrix
 More significant than the superficial and schenckii spores enter
dermatophytoses the skin via abrasions,
scratches, and small
Most Common Infections: puncture wounds as a
result of unprotected
1. Sporotrichosis contact exposure
2. Mycetoma to Sphagnum moss
3. Chromomycosis

SPOROTRICHOSIS

Common Name

 Rose Gardener’s Disease Clinical Manifestations

 Rose Picker’s Disease
 Most cases are cutaneous (relatively local/shallow &
SPOROTHRIX SCHENCKII
mild) or lymphocutaneous
 The only dimorphic fungus to cause subcutaneous Primary “Fixed” Cutaneous  At site of injury are
mycosis Lesions small papules
 Grow’s on thorns of roses (colored raised
 Can be inoculated into the body by an innocent prick area) most often
of the thorn. occurring on an
 The fungus can then morph to a yeast and grow in extremity
the lymph system, manifesting itself as lesions in the
lymph nodes.

Epidemiology

 Occurs worldwide and in all age groups, although it Lymphocutaneous  Lymph nodes
is more common in tropical and subtropical areas become sequentially
 Commonly found in soil and on decaying infected as
vegetation and is a well-known pathogen of organisms are swept
humans and animals. along the lymph
 More infections occur in men due to occupational channels
exposure (foresters, gardeners, horticulturists)  Lymph nodes
become enlarged,
 Sporothrix schenckii grows frequently as an
firm, and discolored
environmental saprobe on woody plants and rich
(buboes)
organic soil  Draining sinuses may
 Well-known to grow on ROSES develop from a lymph
 Many recent cases have been traced to sphagnum node and terminate in
moss purchased from commercial suppliers. the adjacent skin
 On rare occasions
ROSES Most cases are traced disease occurs
to rose thorns, splinters following when
or other plant materials conidia from
penetrating the skin environment are
inhaled (in particular
SPHAGNUM MOSS This is used as a soil the recent cases
conditioner which related to sphagnum
increases the soil's moss exposure)
capacity to hold water
and nutrients by  The rare pulmonary
cases cause
increasing capillary
symptoms that range
from bronchitis to
MYCOLOGY - VIROLOGY RTRMF. MLS 3B
tuberculosis-like o Shoulder and
infections o Arms.

 The most common Clinical Manifestations

extracutaneous
disorder is  Most cases start out as a small hard painless
osteoarticular in nodule which over time begins to soften on the
nature – confined to surface and ulcerate to discharge a viscous,
long bones near joints purulent fluid containing grains.
 The infection slowly spreads to adjacent tissue,
 Dissemination is rare including bone, often causing considerable
and limited to
deformity.
immunocompromised
patients: many  Sinuses continue to discharge serosanguinous
organs involved fluid containing the granules which vary in size,
color and degree of hardness, depending on the
etiologic species.
Morphology: Colonial and Microscopic  GRAINS are the hallmark of mycetoma.

 At 25C, colonies are slow growing, moist and

glabrous, with a wrinkled and folded surface.
 Some strains may produce short aerial hyphae and
pigmentation may vary from white to cream to
black.
 Conidiophores arise at right angles from the thin
septate hyphae and are usually solitary, erect and
tapered towards the apex.
 Conidia are formed in clusters on tiny denticles by Causative Agent
sympodial proliferation of the conidiophore, their
arrangement often suggestive of a flower.  Pseudallescheria boydii / Scedosporium
 As the culture ages, conidia are subsequently formed apiospermum and Scedosporium prolificans
singly along the sides of both conidiophores and  Exophiala jeanselmei
undifferentiated hyphae.  Madurella grisea
 Conidia are ovoid or elongated, 3-6 x 2-3 um in  Madurella mycetomatis
size, hyaline, one-celled and smooth-walled. In  Acremonium sp.***
some isolates, solitary, darkly pigmented, thick-  Aspergillus nidulans***
walled, one-celled, obovate to angular conidia
may also be observed along the hyphae. SCEDOSPORIUM SPP.
 On BHI blood agar at 37C, colonies are glabrous,
 S. apiospermum or S.prolificans in the asexual
white to grayish yellow and yeast-like, consisting
form of P. boydii
of spherical or oval budding yeast cells.
 The two are considered major human pathogens.
Diagnosis
Ecology
 Diagnosis of the disease can sometimes be aided by
The fungi are cosmopolitan and can be isolated from rural
the SPOROTRICHIN SKIN TEST.
soils, polluted water, composts and manure.
 Patients with all forms of the disease develop
Characteristics
circulating antibodies. Although it is not clear whether
these are of any protective value, they do allow for
 The growth of Scedosporium species has been
serological confirmation of the disease.
extensively studied.
MYCETOMA  The ability of these organisms to grow on medium
with cycloheximide is variable, and this factor may
 Is a chronic, suppurative infection of the be used as a means of differentiating S.
subcutaneous tissue and contiguous bone. apiospermum from S. prolificans, which is unable to
 The clinical features are fairly uniform, regardless grow in the presence of this compound.
of the organism involved.  Both species grow optimally between 30ºC to 37ºC
 Most Common Site of Infection: FEET, account for within 48 to 72 hours
at least two-thirds of cases.
Cultural/Colonial Characteristics
 Also referred to as Maduromycosis or Madura Foot
Infection.
 Rate of growth: moderately rapid; mature within 7
 Other sites include the: days
o Lower legshands
 Grayish-white, suede-like to downy with a
o Head
grayish-black reverse
o Neck
o Chest
MYCOLOGY - VIROLOGY
 Both species grow optimally between 30ºC to 37ºC
within 48 to 72 hours
Microscopic Morphology
S. Apiospermum
 Numerous single-celled, pale-brown, broadly
clavate to ovoid conidia, 4-9 x 6-10 mm, rounded
above with truncate bases (cut-off at the base) are
observed.
 Conidia are borne singly or in small groups on
elongate, simple or branched conidiophores or
laterally on hyphae.
 Conidial development can be described as
annellidic, although the annellations (ring-like
scars left at the apex of an annellide after conidial
secession) are extremely difficult to see.
S. prolificans
 Is distinguished from other members of the genus, in
particular, the human pathogen S. apiospermum, by:
 Having basally swollen (inflated), flask-shaped
annellides
 Slower colony development on nutrient agar
media
 By NOT growing on media containing
cycloheximide (actidione).
 Conidia are borne in small groups on distinctive
basally swollen, flask-shaped annellides, which
occur singly or in clusters along the vegetative
hyphae.
 Conidia are single-celled, hyaline to pale-brown,
ovoid to pyriform, 2-5 x 3-13 um (average 3.4-5.3
um) in size, and have smooth thin walls.
RTRMF. MLS 3B
PSEUDASCHELLERIA BOYDII
 Pseudallescheria boydii- sexual state
 Takes priority over the asexual state
name (Scedosporium apiospermum). However,
the sexual state often is NOT present in clinical
specimens and unless it is observed, the fungus
must be referred to by the asexual name
of Scedosporium apiospermum
Macroscopic (Colony)
 Moderately rapid growing colony at 25C which
matures in about a week.
 Texture is woolly to cottony.
 Colony is white becoming greyish or smokey-
brown on the surface; reverse is pale with
brownish-black zones.
 Sexual state P. boydii is inhibited by
cycloheximide (fungal selection agars) while the
asexual state S.apiospermum is NOT inhibited.
Microscopic
 Hyphae are hyaline, septate (2 - 4 µm dia.)
 Conidiophores bearing annellides are of varying
length and exhibit little differentiation from the
vegetative hyphae.
 Conidia (annelloconidia) are unicellular, pale
brown, ovoid with truncate bases formed singly or
in small clusters at the ends of the conidiophores
or from short annellidic necks arising directly from
the hyphae.
 Large (50 to 250 µm) brown cleistiothecia (of the
sexual P.boydii) may develop after 2 to 3 weeks of
incubation. More likely to be found at center of
colony.
 Ascospores are yellow-brown and ellipsoidal in
shape or sperm-shaped.
 Cleistiothecia– sac-like structure with asci and
ascospores.
Laboratory Diagnosis
 Antifungal Susceptibility Testing
 Culture Techniques and Media
 Microscopy Techniques and Stains
MYCOLOGY - VIROLOGY RTRMF. MLS 3B
EXOPHIALA JEANSELMEI

 Rate of Growth: slow; mature within 14 days at 25-

30ºC.
 Grows more slowly or NOT at all at 37ºC
 Cultures grow at 37C but NOT at 40C
 Colonies are initially smooth, greenish-grey to
black, mucoid and yeast-like, becoming raised and
developing tufts of aerial mycelium with age,
often becoming dome-shaped and suede-like in Microscopic Characteristics
texture.
 Reverse is olivaceous-black  Septate hyphae:
o Mostly wide (3-5 um)
o Branched and dark
o Sometimes appear chains of rounded
cells, suggesting a budding process

Microscopic Characteristics

Numerous ellipsoidal, yeast-

like, budding cells are usually  Thinner (1-3 um), cylindric, branched hyphae may
present, especially in young also be present, occasionally with
cultures. chlamydoconidia.
 Grains of Madurella grisea (tissue
Scattered amongst these microcolonies):
yeast-like cells are larger, o Black,
inflated, subglobose to o Round to lobed,
broadly ellipsoidal cells o Soft to firm,
(germinating cells) which give o Up to 1.0 mm, with two distinctive zones,
rise to short torulose hyphae o A hyaline to weakly pigmented central
that gradually change into zone and
unswollen hyphae. o Deeply pigmented periphery.

Conidia are hyaline, smooth,

thin-walled, broadly
ellipsoidal, 3.2-4.4 x 1.2-2.2
µm, and with inconspicuous
basal scars.

MADURELLA GRISEA

Cultural/Colonial Characteristics MADURELLA MYCETOMATIS

 Rate of Growth: Moderately slow at 25-30ºC; Cultural/ ColonialCharacteristics
mature in 12 days; does NOT grow well, if at all at
37ºC  Rate Of Growth: Moderate at 37ºC; mature in 10
 Colonies are slow growing, dark, leathery, folded days; grows much more slowly at 25ºC
with radial grooves and with a light brown to  Colonies vary greatly;
greyish surface mycelium o Maybe smooth or folded,
 With age colonies become dark brown to reddish- o Glabrous or powdery;
brown and have a brownish-black reverse. o Ranges in color from white to yellowish
 Optimum Temperature Of Growth: 30C, it does brown
NOT grow at 37C.  A brown diffusible pigment is PRESENT in the
agar
 Reverse: Dark Brown
MYCOLOGY - VIROLOGY RTRMF. MLS 3B

Microscopic Characteristics
Phialophora Type Clusters of conidia
extrude from
 Two types of Conidiation have been observed, the
flasked-shaped
o First: Flask-shaped phialides that bear
phialides
rounded conidia,
o Second: Simple or branched
conidiophores bearing pyriform conidia (3-5
um) with truncated bases.

Disease

 Chromomycosis / Chromoblastomycosis

Cultural/Colonial Characteristics

 Dematiaceous fungi have similar characteristics.

CHROMOMYCOSIS/ CHROMOBLASTOMYCOSIS  All are slow-growing and produced heaped-up and
slightly folded, darkly pigmented colonies with a
Demaciaceous Fungi (Imperfect Fungi)
greyish-velvety appearance.
Three Genera  Reverse: Jet Black
 Yeast species are usually pleomorphic and
Produce varying amount of melanin-like pigments found in the produce a dark, moist colony.
conidia or hyphae or both.
FONSECAEA SPECIES
Chlasdosporium species C. carrionii
 The genus contains two species, Fonsecaea
Phialophora species P. verrucosa
pedrosoi and Fonsecaea monophora which are
Fonsecaea species F. pedrosoi recognized etiologic agents of
chromoblastomycosis.
F. compacta  Both species are soil fungi and F. pedrosoi is also
associated with forest litter decomposition.

General Characteristics

 Have similar characteristics

 ALL are slow-growing
 Produce Three Types Of Sporulations:

Cladosporium Type Elliptical conidia in

chains, some shield-
shaped due to scars
(disjunctions)

General Charactersitics

 Morphologically the genus Fonsecaea is defined by

the presence of indistinct melanised
conidiophores with blunt, scattered denticles
Rhinocladiella Type Conidia borne singly bearing conidia singly or in short chains that
at end and side of eventually become branched.
conidiophores  de Hoog et al. (2004) revised the genus on the basis
of ribosomal DNA internal transcribed spacer
(ITS) sequence data recognising two species;
o F. pedrosoi and
MYCOLOGY - VIROLOGY
o F. monophora.
 The previously described species F. compacta was
found to be a morphological variant of F. pedrosoi.
 Morphologically F. pedrosoi and F. monophora are
very similar and can best be distinguished by
genetic analysis
 ALL strains grow at 37C but NOT at 40C. Both
species are recognised etiologic agents of
chromoblastomycosis.
 F. pedrosoi is the most common cause of
chromoblastomycosis
 Fonsecaea exhibits in vivo dimorphism; it
produces a specific structure (sclerotic body) only
in tissue and grows in mold form in laboratory
conditions
 Sclerotic bodies are single or clustered, round,
thick-walled, dark-brown to black organisms that
multiply by planate division.
Colonial/Cultural Characteristics
C. carrionii
 Colonies are slow growing, flat to heaped and
folded, suede-like to downy, olivaceous to black
with black reverse.
Characteristics of Clasdosporium
Microscopic Characteristics
 Conidiogenous cells are pale olivaceous,
arranged in loosely branched systems, with
prominent denticles.
Cultural/Colonial Characteristics
 Conidia are pale olivaceous, clavate to
ellipsoidal, in short chains, subhyaline, smooth
and thin-walled, 3.5-5 x 1.5-2 µm.
RTRMF. MLS 3B
 Conidia are 1-celled and arise upon swollen
denticles that are located at the tips of the
conidiophore
 The secondary conidia often produce tertiary
series of conidia like those formed by the primary
conidia, resulting in a complex conidial head.
Microscopic Difference
F. pedrosoi F. monophera
Long conidial chains Slightly longer conidial chains
Short denticles Slightly shorter
PHAEOHYPHOMYCOSIS
Phaeohyphomycotic Fungi
Chlasdosporium Species
 Synonym: Cladophialophora carrionii
 A genus of fungi including some of the most common
indoor and outdoor molds
 Conidiation is of cladosporium type only.
 Species produce olive-green to brown or black
colonies, and have dark-pigmented conidia that
are formed in simple or branching chains.
 Many species of Cladosporium are commonly found
on living and dead plant materials.
 Some species are plant pathogens, other
parasitize other fungi.
 Cladosporium spores are wind-dispersed and they
are often extremely abundant in outdoor air.
 Indoors Cladosporium species may grow on
surfaces when moisture is present.
 Colonies are slow growing, reaching 3-4 cm in
diameter after one month, with a compact suede-
like to downy surface and are olivaceous-black in
color.
MYCOLOGY - VIROLOGY RTRMF. MLS 3B
Microscopic Characteristics Wangiella dermatitidis

 Microscopy shows ascending  A dematiaceous, cosmopolitan saprobic fungus that

to erect, olivaceous-green, inhabits the soil and plant material.
apically branched, elongate  Wangiella species is an occasional agent of various
conidiophores producing infections in humans.
branched chains of smooth-  A neurotropic fungus. Infections in the central
walled conidia. nervous system and as well as cases of keratitis,
otitis, pneumonia, and endocarditis have been
 Conidia are pale olivaceous, reported.
smooth-walled or slightly verrucose to fusiform,  In animals, may cause bovine abortion and still
1.5-3.0 x 2.0-7.0 µm in size. birth.
 Bulbous phialides with large collarettes and  May also cause disseminated infection in
minute, hyaline conidia are occasionally formed on immunocompromised persons
nutritionally poor media.

 Maximum growth temperature 35-37C.

Chladosporium bantianum

 Cladophialophora bantiana may be distinguished

from Cladosporium species by:
1. The absence of conidia with distinctly
pigmented hila,
2. The absence of characteristic shield
cells, and
3. By growth at 42ºC (compared with
Cladophialophora carrionii which has a
maximum growth temperature of 35-
36C).
 Synonyms
o Cladophialophora bantiana,
o Xylohypha bantiana,
o Cladosporium trichoides
Colonial/Cultural Characteristics
 Colonies are moderately fast growing, olivaceous-
grey, suede-like and grow at temperatures up to
42-43C.
Colonial Morphology
 Growth rate is slow and colonies are initially
moist, shiny, yeast – like becoming velvety
around the periphery after incubation at 25°C;
 The surface colony color is black to olive black and
as well as on the reverse;
 Aerial hyphae develop after three to four weeks of
incubation
 Has the ability to grow at high temperatures high
as 42°C and does NOT assimilate p, as potassium
nitrate.
 Colony composed of both budding yeast and
hyphae on Sabouraud glucose agar.

Microscopic Charactertistics

 Conidia are formed in long, sparsely branched,

flexuous, chains from undifferentiated
conidiophores.
 Conidia are one-celled (very occasionally two-
Microscopic Morphology
celled), pale brown, smooth-walled, ellipsoid to
oblong-ellipsoid and are 2-3 x 4-7 um in size.
 Septate brown hyphae,
conidiophores,
phialides, and yeast
cells are PRESENT;
 Brown, budding,
yeast–like cells are
dominant in young
culture when examined
microscopically but as
the culture gets older,
hyphae and phialides
are formed from these
yeast–like cells;
MYCOLOGY - VIROLOGY RTRMF. MLS 3B
 Phialides are brown, branched, flask – shaped to
cylindrical, and without collarettes;
 Conidiophores are commonly difficult to
differentiate from vegetative hyphae; and
 Conidia are unicellular, brown, round - to oval –
shaped, with size ranging from 2 – 4 x 2.5 – 6 µm,
and are found in clusters at the tips of the phialides
and down the sides of the conidiophores;
 Microscopic morphology of Wangiella
dermatitidis showing flask - shaped to cylindrical
phialides without distinctive collarettes or flared-lips.

Phialophora verrucosa

 Is a well-documented
causative agent of
chromoblastomycosis, and
mycetoma.
 It produces characteristic
flask-shaped phialides with
distinctive funnel-shaped,
darkly pigmented
collarettes.
 Environmental isolations have
been made from plant
debris, wood piles, fence
posts, tree stumps, soil and
animal feces.

Colonial Morphology

 Colonies (SDA) are slow growing, initially dome-

shaped, later becoming flat, suede-like and
olivaceous to black in color.

Microscopic Morphology

 Phialides are flask-shaped or elliptical with

distinctive funnel-shaped, darkly pigmented
collarettes.
 Conidia are ellipsoidal, smooth-walled, hyaline,
mostly 3.0-5.0 x 1.5-3.0 μm, and aggregate in slimy
heads at the apices of the phialide.