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MYCOLOGY-VIROLOGY
MIDTERM LECTURE NOTES
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ENABLING OBJECTIVES: At the end of the period,
the students will be able to.
1. Discuss each medically important fungus as to:
Morphology and Physiology
methods of transmission
pathogenesis and clinical manifestations
methods of diagnosis
prevention and control
2. Perform slide preparation of fungal cultures3. Identify a fungus based on gross and microscopicappearance
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CONTENTS
Subcutaneous Mycoses
Sporothrix schenkii Loboa loboi
Agents of Chromomycosis Basidioboulus spp.
Agents of Mycetoma Conidiobolus spp.
Rhinosporidium seeberi
Systemic Mycoses
Histoplasma spp. Blastomyces dermatitidis
Cocciciodes immitis Paracoccidiodes braziliensis
Opportunistic Mycoses
Candida spp.
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SUBCUTANEOUS MYCOSES
Caused by exogenous fungi that normallyreside in nature, mostly in soil and vegetations
Portal of entry
Chronic infections
Sporotrichosis
Mycetoma
Chromoblastomycosis
Phaeohyphomycosis
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Sporotrichosis
Rose gardeners disease
Chronic infection of the subcutaneous tissues and
lymphatics
trauma (thorns or splinters) hand, arm or leg
Occupational hazard
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Sporotrichosis
Clinical manifestations
Fixed cutaneous sporotrichosis
Lymphocutaneous sporotrichosis
Pulmonary sporotrichosis
Osteoarticular sporotrichosis
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Sporotrichosis
Fixed cutaneous sporothricosis
Primary lesion begins as a small, non-healing
ulcer, commonly in the index finger or the
back of the hand
Lymphocutaneous sporotrichosis
nodular lesions
lymphatic vessels and lymph nodes draining the
region
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Sporotrichosis
Sporothrix schenckii
Dimorphic fungi
aspirated pus from nodules, swabs, scrapings,
biopsy tissue
Macroscopic
Rapidly growing, white,
pasty, moist colonythat later becomes
brown, black, wrinkled
or leathery
Microscopic
Mycelial form: narrow,
septate hyphae withpyriform conidia
arranged singly or in a
flowerette
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Mycetoma
Madura foot or Maduromycosis
Traumatic inoculation with several saprophytic
fungi
lower extremities but may occur in any part
of the body
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Mycetoma: Types
1. Actinomycotic
(bacterial)
Actinomycetes
Actinomyces Nocardia
Streptomyces
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MYCETOMA
2. Eumycotic (fungal)
Pseudallescheria boydii most common
Acremonium falciforme
Exophiala jeanselmei Curvularia
Madurella mycetomatis
Madurella grisea
Black grainmycetoma
Whitegrainmycetoma
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Pseudallescheria boydii
Ascomycota group
Soil, standing water and sewage
Clinical specimens: granules from the lesions Pseudoallescheriasis
Meningitis
Arthritis
Endocarditis
Brain abscess
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Pseudallescheria boydii
Macroscopic
Rapidly growing (5-10
days), initial growth as a
white fluffy colony
after several wks to
brownish-gray colony
Reverse tan to dark
brown
Microscopic
Asexual form :
Scedosporium apiospermum
golden brown elliptic,single-celled conidia borne
singly from the tips of
conidiophores
Sexual form : brown sac-like
cleistothecia containing asci
and ascospores
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Chromoblastomycosis(Chromomycosis)
Traumatic inoculation
Chronic infection producing warty or
cauliflower-like or tumor-like lesions mostly in
the lower extremities
Epidermis hyperplasia
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Chromoblastomycosis(Chromomycosis)
Etiologic agents
Cladosporium (Cladophialophora carrionii)
Phialophora (Phialophora verrucosa)
Fonsecaea (F. pedrosoi, F. compacta)
Rhinocladiella aquaspersa
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Chromoblastomycosis
Macroscopic All grow slowly and produce heaped-up and slightly folded,
darkly pigmented colonies with a gray to olive to black
velvety colonies; reverse side of colonies is jet black
Microscopic
Cladosporium: chains of budding blastoconidia borne from
branching conidiophores
Phialophora: short flask-shaped phialides with collarette Fonsecaea: conidial heads with sympodial arrangement of
conidia, primary conidia giving rise to secondary or tertiary
conidia
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Fonsecaea
F. pedrosoi
Polymorphic
1. Phialides
2. Chains of blastoconidia
3. sympodial
F. compacta
Spherical w/ broad base
connecting the conidia
Smaller and more
compact than pedrosoi
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Rhinocladiella
Produces lateral or terminal condia from
conidiogenous cell ( sympodial)
Conidia are elliptical to clavate
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Phaeohyphomycosis
Caused by dematiaceous fungi other than
those causing chromomycosis
Tissue morphology is mycelial
Subcutaneous and systemic infection
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Clinical manifestation
Subcutaneous phaeohyphomycosis
Cystic lesion, abscess
Paranasal sinus phaeohyphomycosis
sinusitis
Cerebral phaeohyphomycosis
Immunosuppressed
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PhaeohyphomycosisEtiologic Agents
Exophiala jeanselmei
Wangiella dermatitidis
Phialophora richardsiae Alternaria spp
Bipolaris spicifera
Curvularia spp
subQ: exophiala and wangiellaparanasal sinusitis ( allergic rhinitis or immunosuppression) : Bipolaris, Exserohilum, Curvularia andAlternaria
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Exophiala
Macroscopic
Grow slowly (7-21 days)and initially grows black
yeast-like colonies; as
colonies age they
become filamentous,
velvety, gray to black
Microscopic
Pale brownconidiophores that form
cylindrical annellids,
hyaline conidia gather at
its tip
Wangiella dermatitidis
Subcutaneous phaeohyphomycosis
cystic lesions occur most often in adults : subcutaneous phaeomycotic cyst
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Alternaria
Macroscopic
Grow rapidly and appear fluffy, gray to graybrown or gray green colonies
Microscopic Hyphae: septated and golden brown,
Conidiophores: simple sometimes branched whichbear a chain of large brown conidia resembling a
drumstick
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Curvularia
Macroscopic
Rapid growing, most are fluffy, gray to black
colonies
Microscopic
Hyphae are dematiaceous and septate
conidiophores are twisted at the ends where
conidia are attached
conidia are multicelled, curved with a central
swollen cell
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SYSTEMIC & OPPORTUNISTIC MYCOSES
Primary Systemic Mycoses
Coccidioidomycosis
Histoplasmosis
Blastomycosis
Paracoccidioidomycosis
Opportunistic Mycoses
Candidiasis, systemicCryptococcosis
Aspergillosis
Mucormycosis
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Primary Systemic Mycoses
Caused by dimorphic fungi
Dimorphic fungi
Yeast phase
When grown on enriched media usually supplementedwith blood at 35-37C
Is observed in vivo and is also known as the tissue orinvasive phase
Mycelial phase Observed on SDA at 25-30C
Saprophytic, observed in vitro
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Transmission
Inhalation of fungal spores
Lead initially to pulmonary infection which
may be symptomatic or asymptomatic
Dissemination to other body sites can occur
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Coccidioidomycosis
Acquired through inhalation of the infectivearthroconidia
Approximately 60% are asymptomatic and
self-limited respiratory tract infections Infection may become disseminated to
visceral organs, meninges, bone, skin, lymph
nodes and subcutaneous tissue
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Coccidioidomycosis
Etiologic agent: Coccidioides immitis
Clinical specimens: Sputum, tissues or body fluids
Direct microscopic examination from clinicalspecimens
Non-budding, thick-walled spherule, 20-200um in diameter containing either granular
material or numerous small non-buddingspores
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Coccidioides immitis
Macroscopic
Colonies appear after 3-21 days, delicate fluffy whitewhich turn tan or brown with age
Microscopic Mycelial phase: septate, branched hyphae that produce
thick-walled barrel-shaped, rectangular arthroconidia that
alternate with empty alternate cells
Yeast phase: large, round, thick-walled spherules withendospores observed in tissues and direct examination
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Coccidioides immitis
Other nonvirulent fungi that resemble C. immitis
microscopically may be found in the environment andmay produce hyphae that may dissociate into
arthroconidia
Considered as the most infectious of all fungi Extreme caution should be observed in handling
cultures of this organism
If culture plates are used, they should be handled
only in a biological safety cabinet
Cultures should be sealed in tape if the specimen is
suspected of containing C. immitis
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Safety Precautions in Handling
C. immitis Cultures
cotton-plugged tubes is discouraged andscrew-capped tubes are preferred
All microscopic preparations for examination
should be performed inside a BSC Cultures should be autoclaved as soon as the
final identification of C. immitis is made
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Histoplasmosis
A chronic granulomatous infection that is primary
and begins in the lungs, produce cavitary lesions
disseminate to the lymph node, liver, spleen, bone
marrow, kidneys, meninges Heart infxn in immunocompromised indls
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Histoplasmosis
inhalation of conidia or small hyphal
fragments
95% are asymptomatic and self-limited
most prevalent pulmonary mycosis of humans
and animals
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Histoplasma capsulatum
Direct microscopic examination
Difficult to visualize in the sputum and other
tissues
bone marrow smear: Wright or Giemsa-stained
Rarely in peripheral blood
Intracellular yeast in macrophages
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Histoplasma capsulatum
Macroscopic Slow growing
SDA:
white to brown mold
with fine fluffy texture
reverse side: white,
yellow or tan
BHI
moist, white to cream
heaped colony
Microscopic Mycelial phase:
septate hyphae with
large spherical or
pyriform tuberculatemacroconidia; some
produce small round
smooth microconidia
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Blastomycosis
Chronic suppurative and granulomatous
infection which involve the lungs and spread
to the long bones, soft tissue and skin
inhalation of the conidia and hyphal
fragments
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Blastomyces dermatitidis
Direct microscopic examination of tissues or body
fluids
large, spherical, thick-walled yeast cells 8-15 u usually with
a single bud that is connected to its parent cell by a broad
base
Mycelial phase: delicate, septate hyphae with round or
pyriform conidia borne singly on conidiophores resembling
lollipops
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Blastomyces dermatitidis
Macroscopic
Growth rate is 7-21 days
SDA: colony at first white, waxy, yeast-like and later
becoming cottony with white aerial mycelium; turnstan to brown with age
BHI with blood: cream to tan, waxy. Wrinkled colonies
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Paracoccidioidomycosis
Chronic granulomatous infection that begins
as a primary pulmonary infection
asymptomatic but may disseminate to
produce ulcerative lesions
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Paracoccidioides brasiliensis
Macroscopic:
SDA: white, glabrous, leathery colony which turns
tan-brown with age
BA: cream to tan, moist, wrinkled colony whichturns waxy with age
Microscopic
Mycelial phase small, septate, branched hyphae with intercalary
and terminal chlamydospores
few pyriform microconidia
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Paracoccidioides brasiliensis
Yeast phase
large, round to oval,
thick-walled yeast
cells (8-40 u) withmultiple buds with a
narrow base
mariners wheel
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Candidiasis
Most frequently encountered opportunistic
fungal infection
Etiologic agents
Candida albicans
C. tropicalis
C. parapsilosis
C. glabrata
Opportunistic Mycoses
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Candidiasis
are part of the normal flora, seen in the
oropharynx, GIT, GUT, skin
Infections are believed to be endogenous in
origin or nosocomial
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Candida Infections In Normal And
Immunocompromised Hosts
Intertriginous candidiasis (skin folds)
Onychomychosis and paronychia
Perleche
Oral thrush
Vulvovaginitis
Pulmonary infection
Eye infections
Endocarditis
Meningitis
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Predisposing Factors For Candidiasis
Alteration in the normal skin and mucous
membrane barriers
Prolonged antibiotic administration
Use of immunosuppressive drugs
Diseases of the immune system
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Candida
Direct microscopic examination of clinicalspecimens
Budding yeast cells
Pseudohyphae
Definitely identified microscopically by production of
germ tubes and chlamydospores
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Germ Tube Test
A hypha-like extension of the yeast cells with
no constriction at the point of origin
Candida albicans will form germ tubes when
incubated with serum at 37C for a few hours
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Cornmeal Agar with Tween 80
Conidiation
ID of Candida spp and other yeasts through
examination of
hyphae, blastoconidia, chlamydospores. and
arthroconidia
Tween 80
reduce the surface tension
to allow conidiation
C l A ith T 80
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Cornmeal Agar with Tween 80Procedure
colony from the 1 culture media
Inoculate a plate of CMA with 1% T80 and trypanblue by making 3 parallel cuts about inch apart at a45 angle to the culture medium
RT for 48 hours
After 48 hours, remove and examine the areas where
cuts into the agar were made
Commonly encountered yeast in CMA-T80 Agar
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Commonly encountered yeast in CMA T80 Agar
Organism Arthro-
conidia
Blastoconidia Pseudohyphae or
Hyphae
C. albicans
-
Spherical clusters at
regular intervals on
pseudohyphae
Chlamydoconidia
on hyphae
C. glabrata-
Small, spherical, tightlycompact
None
C. krusei
-
Elongated, clusterered
at septae of
pseudohyphae
Branched
pseudohyphae
C. parapsilosis
-
Present but not
characteristic
Sagebrush like,
Giant hyphae
Commonly encountered yeast in CMA-T80 Agar
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Organism Arthro- conidia Blastoconidia Pseudohyphae or
Hyphae
C. kefyr
(pseudotropicalis)- Elongated, parallel to
pseudohyphae
PH present, not
characteristic
C. tropicalis - Randomly appear on PH
& H
PH present, not
characteristic
-
C. neoformans - Round to oval separated
by capsule
Rare, usually not
seen
Saccharomyces - Large and spherical Rudimentary H
sometimes present
Trichosporon Numerous,
resemble
Geotrichum
Maybe present but
difficult to find
Septated hyphae is
present
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Cryptococcosis
An acute, subacute or chronic fungal infectionthat has several manifestations
Disseminated disease
with or without meningitis in immunocompromisedpatients
Meningitis occur 2/3 of patients
very common in patients with AIDS
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Cryptococcus neoformans
Saprophyte
pigeon, bat, or bird droppings, decaying
vegetations, fruit, plants
Inhalation
lungs then disseminate to meninges and other
sites
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Cryptococcus neoformans
Direct microscopic
examination
Spherical, single or multiple
budding, thick-walled yeast
cell (2 to 15 um)
surrounded by a wide,
refractile polysaccharide
capsule
Macroscopic Colonies appear in 1-5
days
smooth, white to tan,
mucoid, gelatin-likecolonies (soap-bubble)
Brown-black colonies
on Niger seed agar
O i C l G Bl t A th Chl
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Organism Capsule Germ
Tube
Blasto-
conidia
Arthro-
conidia
Chlamy-
dospore
C. albicans - + + - +
C. tropicalis - - + - V
C. parapsilosis - - + - -
C. glabrata - - + - -
C. neoformans + - + - -
Geotrichum - - - + -
T. beigilii - - + + -
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Organism FERMENTATION Urease Nitrate
Reduction
G M S L
C. albicans + + - - - -
C. tropicalis + + + - - -
C. parapsilosis + - - - - -
C. glabrata + - - - - -
C. neoformans - - - - + -
Geotrichum - - - - - -
T. beigilii - - - - + -
Aspergillosis
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Aspergillosis
disseminated infection in IC patients
Other infection
invasive lung infection
Pulmonary or sinus fungus ball (tangled mass of hyphae)
Mycotic keratitis
allergic pulmonary aspergillosis
External otomycosis
Onychomycosis
Sinusitis, endocarditis, CNS infxn
inhalation
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Aspergillus fumigatus
Direct microscopic examination
Septate hyphae that usually show dichotomous
branching (45 angle branching)
Macroscopic Rapidly growing mold (2-6 days)
fluffy to granular, white to blue green colonies
Microscopic Branching septate hyphae that terminate in
conidiophore
MOST COMMONLY RECOVERED SPP FROM IC PATIENTA. FLAVUS SOMETIMES RECOVEREDBotany repeated branching into two equal parts.Methenamine silver stained tissue section showing dichotomously branched, septate hyphae.
Aspergillus fumigatus
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Aspergillus fumigatus
expands into a largeDOME-SHAPED vesicle
with BOTTLE-SHAPED
phialides from which
chains of conidia arise
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Zygomycosis (Mucormycosis)
Decaying vegetable matter, old bread or in soil
Acquired by inhalation
Less common cause of infection as compared to
Aspergillus Rhinocerebral infection involving nasal mucosa,
palate, sinuses and brain
Perineural invasion
Retro-orbital spread (brain)
Lungs, GIT
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Zygomycetes
Direct microscopic examination of tissue
specimens or exudates
Branching non-septate hyphae
Macroscopic
Fluffy, white to gray to brown colonies covering
the surface of the agar within 24-95 hours, grayish
hyphae with brown to black sporangia
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Zygomycetes
Microscopic large ribbon-like hyphae
irregular in diameter
non-septate
Sporangia
Sac-like sporangiospores at the tip of sporangiophore
Stolons
Connects sporangiphore
Rhizoids are attached
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Mucor
Sporangiophores the tip
of which have sporangia
filled with
sporangiospores No rhizoids and stolons
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Rhizopus
unbranched
sporangiophores with
rhizoids that appear at
the point at which thestolon arises
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Penicillium
When clinically
significant, clinical
manifestations include
bronchopulmonary,
endocarditis, cutaneous
ulcers of extremities
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Fusarium
Infections becoming more common esp in IC patients
(Hyalohyphomycosis)
Common environmental flora
mycotic keratitis after traumatic implantation into the cornea
Other infections: sinusitis, wound (burn) infections, allergic
fungal sinusitis, respiratory tract secretions
http://www.mycology.adelaide.edu.au/Mycoses/Opportunistic/Hyalohyphomycosis/index.htmlhttp://www.mycology.adelaide.edu.au/Mycoses/Opportunistic/Hyalohyphomycosis/index.html8/13/2019 Midterm Subcut-sys-opportunistic
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Pneumocystis jiroveci (carinii)
Opportunistic atypical fungus causing
pneumonia in immunocompromised hosts
Ideal specimen broncho-alveolar lavage fluid
or lung biopsy
Does not grow in routine culture methods
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Polyene macrolide antifungals
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Agent Source Function Treatment for
Amphotericin B
(liposomal prep)
Streptomyces
nodosus
Binds ergosterol and
alter selectivepermeability
IV:Aspergilossis
Candida spp.Cryptococcus
Zygomycetes
R: P.boydii,
A. terreus,
Trischosporon,Fusarium
Nystatin S. noursei Not absorbed by GIT,
not given
parenterally (TOXIC)
Oral or
vulvovaginal
candidiasis
Griseofulvin Penicillium Binds microtubular
protein (mitosis)
Oral tx:
dermatophytes
non responsive to
azole
Antimetabolite
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Antimetabolite
5- Fluorocytosine (Flucytosine)
5-fluorouracil
incorporated to fungal RNA and inhibit protein
synthesis
Fluorodeoxyuridine monophosphate
Inhibitor of DNA synthesis
Combination therapy w/ AmB
Candida spp. and C. neoformans
Side effect & resistance when used alone
Azole antifungal drugs
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Agent Application Treatment Adverse rxn
Clotrimazole &
Miconazole
Topical or
intravaginal
Mild dermatophytosis
(T.versicolor)
Burning, itching,
skin irritation
Fluconazole Oral or IV Candida and Cryptococcus
(CNS)
S or R (C. glabrata)
R: C.krusei &
Rhodotorula spp)
Ketoconazole Topical or
oral
Mild
ParacoccidioidomycosisBla
stomyces & Histoplasmosis
Chronic mucocutaneous
candidiasisP. boydii
Elevated liver
enzymes, nausea,
dose related-
gynecomastia;
Decreased libido;oligospermia
A l if l d
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Azole antifungal drugs
Agent Application Treatment Adverse rxnItraconazole Expanded
activity w/
ketoconazole
Aspergillosis
Sporothricosis
Cryptococcosis
OnchymycosisBlastomycosis
GIT & vestibular
disturbances,
edema, skin
irritation
Voriconazole
(new triazole)
Expanded
activity
compared
w/itraconazole
Fusarium
C.krusei & C.
glabrata
R: Zygomycetes;
Elev.liver
enzymes; visual
disturbances
E hi di
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Echinocandins
Agent Application Treatment Adverse rxnCaspofungin
Micafungin
Anidulafungin
Fungicidal
Fungistatic
Candida spp
(krusei,glabrata)
Aspergillus
R:
C.neoformans
Trichosporon,
RhodotorulaZygomycetes
Selenium
sulfide
Shampoo
Sporicidal
Malasezzia furfur
T. tonsurans
Potassium
iodide
oral Cutaneous/lymp
hatic
sporothricosis
Bitter taste,
allergic rash
and anorexia
Recommended