MYCOLOGY 2 BASIC TYPES OF PROPAGULES

1. Asexual Sporulation/Propagules
Ø Termed either spores or conidia; Major method for the maintenance & dissemination of many fungi
Plantsà eukaryotic & autotrophic Fungi & Animals à Eukaryotic & heterotrophic Ø Conidia arise either by: budding off conidiogenous hyphae OR by differentiation of preformed hyphae (sporulation)
Protozoa à eukaryotic Bacteria à prokareutic Ø Asexual spores: commonly formed by consecutive cleavages of a sporangium (fragmentation)
A. Simplest Sporulation: development of spore directly from the aerial hyphae
FUNGI (1) Chlamydospore
ð Round thick walled resistant spore formed directly from the differentiation of the hyphae à
• Eukaryotic produced by rounding up and enlargement of terminal cells of the hyphae
- Contain membrane bound cell organelles; also exhibit mitosis (2) Conidia
• Heterotrophic ð Produced singly/in groups by a specialized hyphae à conidiophore
- Lack chlorophyll & are therefore not autotrophic (photosynthetic) ð some conidiophores terminate in a swollen structure à vesicle à from
the surface of the vesicle are formed secondary small flask shaped
- Either saprophytes (living on dead organic matter) or parasites (utilizing living tissue)
phialides which give rise to long chain of conidia (ex: Aspergillus)
• Rigid cell walls à non-motile ð from conidiophores, phialides with secondary branches are formed
• Cell wall: primarily composed of chitin (metulae) followed by a chain of conidia (ex: Penicillium)
• Cell membrane: contains sterol ; Size: varies from visible mushroom to microscopic Candida 2 sizes of conidia:
ð
• Exist in 2 forms (Dimorphism): May be unicellular (yeast) or multicellular (molds) a. Microconidia à small, unicellular round, elliptical or pyriform in shape
b. Macroconidia à large, multiseptate, club-shaped or spindle shaped
What is “dematiaceous”? Dematiaceous fungi- Black Molds (difficult to ID, numerous sp.) (source: net) (3) Sporangiospore
ð Spores found inside a large sac-like structure called sporangium,
Characteristics of Fungi supported by a nonseptate hyphae called sporangiophore. Ex:
Zygomycetes (Mucor)
Aerobic Susceptible to polyene antifungal agents B. Budding
Grow best at room temp All reproduce asexually; some may reproduce ð Blastoconidia: found in yeast cells
Grow in presence of acid & large amount of sugar sexually ð Pseudohyphae: blastoconidia may elongate & remain attached
(SDA pH of 5.6) Resistant to Cold C. Fragmentation Hyphae
Resistant to antibacterial antibiotics Easily killed at high temp ð Arthroconidia: hyphae fragments at septa
ð Appears as square, rectangular or barrel-shaped thick walled cells
Structure (basic growth forms/stages) 2. Sexual Propagules
Yeast Filamentous / Mold Ø Production of specialized structures from nuclear fusion
Ø Produced by the fusion of two nuclei that generally undergo meiosis
- Reproduces by simple budding to form - A vegetative growth of filaments Ø Sexual methods of reproduction involve:
blastoconidia - Reproduction: spores/conidia ð Plasmogamy: cytoplasmic fusion of 2 cells ; Karyogamy: fusion of 3 nuclei ; Genetic recombination & meiosis
- Colonies: usually moist/mucoid - Produce a great variety of conidia which are borne Ø resulting haploid spore is said to be a sexual spore:
- May be basidiomycetes, such as Cyptococcus on specialized hyphae or conidiophores Zygospore - tips of approximating hyphae fuse à meiosis occurs à large thick walled zygozpores
neoformans - ID by: morphology of spores & by their arrangement Ascospores -4-8 spores within a specialized cell called ascus
- Or ascomycetes such as Candida albicans on the hyphae Basidiospores- 4 spores usually form on the surface of specialized cells called a Basidium
Ø heterothallic: a sexual spore produced by fusion of a nucleus of one mating type with a nucleus of another mating type
-Unicellular -Formed by production of multicellular branching Ø homothallic: a sexual spore produced following fusion of 2 nuclei from the same strain
-Spherical to ellipsoidal cylindrical tubules called hyphae Laboratory Diagnosis
-Reproduction: -Septate Hyphae- with cross walls
(usually) budding -Aseptate- Order Mucorales 1. Direct microscopic examination
(few) binary fission -Mycelium- mass of intertwining hyphae formed during - Wet mount
Few buds fail to detach producing a chain of active growth ð KOH wet mount (10-20% KOH)
elongation yeasts cells- Pseudohyphae -Vegetative/ Substrate mycelium -Mechanism: KOH separates the fungal elements from intact cells as it digests the protein
-Aerial/ reprofuctive mycelium debris and dissolves cement substances that holds the keratinized cells together and makes it
easy to visualize under the microscope. (net)
Dimorphism -Improved sensitivity with calcouflour white- makes it fluorescent
ð India ink wet mount ; Lactophenol cotton blue wet mount
• The ability of some fungi to grow in more than one form under different environmental conditions - Staining
- Yeast form: 37 celcius Mold form: room temp ð Gram staining ; H & E (formalin fixed)
Reproduction: ð Special fungal cell wall stains:
o Gomori- methenamin silver (GMS) , Periodic Acid Schiff (PAS) , Capsule stain
Fungal Spores Asexual Spores o India Ink- A diagnostic test used to detect the cryptococcal organism C. neoformans
ð Wright or giemsa staining - for H. capsulatum in the macrophages
- Anamorphic: derived from asexual repro - Conidia
2. Culture media
- Teleomorphic: from sexual repro - Sporangiospore: Order Mucorales - SDA: glucose and modified peptone (pH7)
with or w/o antibacterial antibiotics (chloramphenicol) and cycloheximide
Hyphae - Incubate at room temperature for 30 days before reporting as negative
- Microscopic exam of cultures:
• Hyphae: fungal elements Mycelium: collective mass of hyphae
ð Lactophenol cotton blue wet mount
• Two kind of hyphae:
ð Slide culture – demonstrate undisturbed morphology of fungus
1. Non-septate/coenocytic à Zygomycetes
3. Blood Culture
2. Septate à septa divide hyphae into compartments; has pores; ex: Basidiomycetes & Ascomycetes
4. Biochemical Tests
5. Serology
- Agglutination: CALAS: Crytococcal Antigen Latex Agglutination System ; Precipitation
(NFT 2021) Info in RED= Dr. Chua’s lecture 2019; black= from medical mycology.doc 6. Molecular Methods
- PCR and MALDI- TOF -MS: extraction of microbial proteins then mass spectroscopy
MAJOR FUNGAL INFEECTIONS (Degree of tissie involvement and mode of entry)

Type of Infection Causative organism Disease Type of Infection Causative organism Disease
Superficial Malassezia species Pithriasis versicolor Blastomyces dermatitis
Blastomycosis
Trichophyton Dermatophytosis Histoplasma capsulatum
Endemic Histoplasmosis
Epidermophyton Coccidioides immitis
Cutaneous (primary systemic) Coccidioidomycosis
Macrosporum Paracoccidioides
Paracoccidioidomycosis
Candida albicans Candidiasis bransiliensis
Aspergillus fumigatus Aspergillosis
Candida albicans Candidiasis
Subcutaneous Sporothrix schenkii Sporotrichosis Opportunistic
Cryptococcus neoformans Cryptococcosis
Mucor/Rhizopus Zygomycosis

CLASSIFICATION OF FUNGI

• Primarily based on the method of spore production of the perfect/sexual state(teleomorph)

• 3 major subdivisions: Zygomycota, Ascomycotina and Basidiomycotina
• Deuteromycotina: accommodates those fungi without a perfect state, but which may represent the asexual states (anamorphs) of either Basidiomycotina or Ascomycotina

Zygomycetes (Bread or Pin Moulds) Hyphomycetes (Conidial Moulds; considered Deuteromycotina)

• Fast growing, terrestrial, largely saprobic fungi with non motile cells • Reproduce asexually by conidia on hyphae or aggregations of hyphae but not with in discrete conidiomata
• Cosmopolitan (665 species) • No sexual state; cosmopolitan (17,000 species)
• Hyphae coenocytic & mostly aseptate • Hyphae: septate with simple ascomycetous septal spores
• Asexual spores include: • Contains the majority of medically important fungi
- Chlamydoconidia 1. Dematiaceous hyphomycetes
- Conidia ð Conidial fungi that produce dark brown, green-black, or black colonies and are the causative
- Sporangiospores contained in sporangia borne on simple or branched sporangiophores agents of phaeohyphomycosis
• Sexual production is isogamous producing a thick-walled sexual resting spore called a zygospore 2. Hyaline hyphomycetes
• However, most isolates are heterothallic, i.e. zygospores are absent, therefore ID is based primarily on sporangial morphology ð Conidial fungi which are not darkly pigmented; colonies may be colourless or brightly
• Medically important Zygomycetes coloured
- Entomophthorales subcutaneous zygomycosis (Entomophthoromycosis) ð Include agents of: hyalohyphomycosis, aspergillosis, dermatophytosis and the dimorphic
ð Conidiobolus & Basidiobolus pathogens like Histoplasma capsulatum
- Mucorales: subcutaneous and systemic zygomycosis (Mucormycosis) • ID:
ð Rhizopus - Primarily: Microscopic morphology of the conidia, their arrangement, and on what type of cell
ð Mucor produces them
ð Rhizomucor - Culture characteristics: less reliable but may also be useful (include surface texture, topography &
ð Absidia, Cunninghamella, Mortierella, Saksenaea & Apophysomyces pigmentation, reverse pigmentation & growth at 37 celcius)

Basidiomycetes (Mushrooms & Toadstools) Ascomycetes (common moulds)

• Saprobes and parasites (esp of plants), terrestrial; cosmopolitan (16,000 species)

• Hyphae septate with complex septal pores called dolipores which allow cytoplasmic (but not nuclear) migration
• Saprobes, parasites (esp of plants), or lichen forming, mostly terrestrial; cosmopolitan (28,650 species)
• Can often be distinguished by the presence of clamp connections over the septa
• Hyphae septate with simple septal spores, cytoplasmic and nuclear migration not inhibited
• Sexual reproduction is by the formation of exogenous basidiospores, typically four, on a basidium
• Asexual reproduction is by conidia
• Occasional species produce conidia but most are sterile
• Medically important genera:
• Genera of Medical importance:
- Teleomorphs of known pathogenic fungi
- Teleomorphs of known pathogenic fungi
ð E.g. Arthroderma, Nannizzia, Ajellomyces, Pseudallescheria, Eurotium etc.,
- Coprinus & Schizophyllium agents of basidiomycosis
- Agents of mycetoma, like Leptosphaeria & Neotestudina
- Mushroom poisoning by Aminita, Lepiota, Coprinus and Psilocybe etc.
- Agent of black piedra, like Piedraia hortae

Fungal Pathogenecity

• The ability to cause disease appears to be an accidental phenomenon • There are then many “opportunistic” fungi which cause
• With the exception of a few dermatophytes, pathogenicity among the fungi is NOT necessary for the maintenance or dissemination of the species infections almost exclusively in debilitated patients
• In general, the devt of human mycoses is related primarily to the ff rather than the infecting organism: whose normal defense mechanisms are impaired
1. Immunological status of the host
2. Environmental exposure Clinical Groupings for Fungal Infections
• Basic mechanism of fungal pathogenicity is its ability to:
SKIN MYCOLOGY INFECTIOUS DISEASE
1. Adapt to the tissue environment Superficial Mycoses MYCOLOGY
2. Withstand the lytic activity of the host’s cellular defenses Cutaneous Mycoses Dimorphic Systemic Mycoses
• A small number of fungi have the ability to cause infections in normal healthy humans by: Subcutaneous Mycoses Opportunistic Systemic
1. unique enzymatic capacity Mycoses
2. thermal dimorphism
3. Ability to block the cell-mediated immune defenses of the host
SUPERFICIAL MYCOSES

• Superficial cosmetic fungal infection of the skin or hair shaft 1. Tinea versicolor (Pityriasis versicolor)
• No living tissue is invaded and there is no cellular response from the host à no pathological changes elicited 2. Black Piedra
• Often so innocuousà Px often unaware of their condition 3. White Piedra
• Affects only the cornified layers of the epidermis and suprafollilcular portion of the hair. 4. Tinea Nigra
• Do not infect the stratum granulosum and the nails

1. Pityriasis versicolor 2. Black Piedra

ð A chronic, superficial fungal disease of the skin characterized by well-demarcated white, pink, ð A superficial fungal infection of the hair shaft cause by Piedra hortae, an
fawn or brownish lesions; often coalescing; covered with thin furfuraceous scales ascomycetous fungus forming hard black nodules on the shafts of the
ð Colour varies accdg to: scalp, beard, moustache & pubic hair
o Normal pigmentation of Px ; Exposure of the area to sunlight; & Severity of the ð Mostly affects young adults & epidemics in families have been reported
following the sharing of combs & hairbrushes
disease
ð Etiology: Piedra hortae
ð Lesions occur on:
o Dark walled mycelium, spreads over & around the hair shaft
o Trunk. Shoulders, Arm
à hard & grittly nodule
o Rarely: neck & face
o Nodules: made up of a mass of pigmented w/ a stroma-like
ð Wood’s ultra-violet light: fluoresce a pale greenish colour center containing asci
ð Young adults affected most often; but disease may occur in childhood & old age ð Diagnosis
ð Distribution: worldwide; more common in tropical than temperate climates (1) Direct Examination
ð Etiologic agent: Malassezia furfur o 10% KOH wet mount will show septated dematiaceous
o A lipophilic yeast forming part of the normal flora of human skin mycelium in the edges of the nodule
o Causative agent also of Pityriasis folliculitis o Crush nodule in 10% KOH à oval asci w/ 8 aseptate
o Has recently been implicated as a causative agent of seborrhoeic dermatitis & curved, spindle shaped ascospores w/ filament at each
dandruff pole
ð Laboratory Diagnosis: (2) Culture
(1) Direct Examination: skin scrapings taken from Px with Pityriasis versicolor stained rapidly o SDA w/ antibiotics
when mounted in 10% KOH, glycerol & Parker ink solution o Inhibited by cycloheximide
o Colonies: black to dark brown, heaped, grabrous,
o Characteristic clusters of thick-walled, round, budding yeast-like cells & short
covered w/ fine downy mycelium. Hyphae –
angular hyphal forms (spaghetti w/ meatballs)
dematiaceous. Fails to produce asci & ascospores
(2) Culture on SDA overlaid w/ sterile olive oil or lanolin + antibiotic to prevent contamination
à not necessary (KOH wet mount is diagnostic)
(3) Woods Lamp
3. White Piedra 4. Tinea Nigra
ð A superficial cosmetic fungal infection of the hair shaft ð A chronic & asymptomatic infection of the stratum corneum manifested
ð Usually localized to the axilla or scalp but may also be seen on facial hairs & sometimes pubic hair by the devt of blackish-brown macules on the smooth skin of the body
ð Common in young adults (usually on the palms of the hands)
ð Presence of irregular, soft, whote or light brown nodules, 1.0-1.5 mm in length, firmly adhering to the ð Etiology: Hortaea werneckii (old name: Exophiala werneckii)
hairs à characteristic o Culture of scrapings from lesion
ð Etiology: Trichosporon beigelii (Trichosporon beigelii/T.cutaneum) o SDA w/ and w/out antibiotics
ð Diagnosis: ê Colony: initially shiny, moist & often white to gray in color
(1) Direct examination: à few days later à olive or black in color à later
o look for irregular, soft, white to light brown nodules, 1.0-1.5 mm in length, firmly mycelium develops & colony appears dull & fuzzy
adhering to the hairs. ê Microscopic exam of colony
o 10% KOH wet mount of crushed soft nodule à hyaline mycelium & arthroconidia = young colony: budding yeast cells, in chain
(2) Culture = older: mixture of hyphae & yeast cells
o SDA with antibioticsw/o cycloheximide ð Diagnosis:
o Rapid growth à cream colored yeast-like colonies o Direct Examination à 10% KOH wet mount of scrapings from
o Microscopic examination: blastoconidia in chain/clusters, arthroconidia periphery of lesion à pigmented brown to dark olivaceous
(dematiaceous) septate hyphal elements and 2-celled yeast cells
producing annelloconidia
CUTANEOUS MYCOSES

Disease Causative organisms Incidence

Dermatophytotis Dermatophytes
• Most widely distributed à invade keratinixed areas of the
fungal disease bodies- skin hair and nails
Ringworm of the scalp, glabrous skin à utilize keratin as source of nutrient
and nails
Candidiasis of the skin, mucous Candida albicans and related species
membranes and nails
Dermatpmycosis Non-dermatophyte moulds
Hendersonula toruloidea
Scytalidium hyalium
Scopulariopsis brevicaulis
Microsporum- M. audounii
Trichophyton- T. tonsurans
Epidermophyton- E. floccosum
Common
Common • Are superficial fungal infections of the skin, hair or nails
• No living tissue is invaded
Rare • However a variety of pathological changes occur in the host
because of the presence of the infectious agent and its metabolic
products.

Dermatophytosis – Ringworm or Tinea

o Ringworm of scalp, glabrous skin, and nails caused by dermatophytes
§ Have the ability to utilize keratin as a nutrient source, i.e. they have a unique enzymatic capacity – keratinase
o Unique for two reasons:
§ No living tissue is invaded
- The keratinized stratum corneum is simply colonized
- Presence of the fungus and its metabolic products usually induces an allergic and inflammatory eczematous response in the host
- The type and severity of the host response is often related to the species and strain of dermatophyte causing the infection
§ Are the only fungi that have evolved a dependency on human or animal infection for the survival and dissemination of their species
o 3 broad epidemiological groups of dermatophytes

Geophilic Zoophilic Anthropophilic

Normally inhabit the soil where they are
believed to decompose keratinaceous debris
Some species may cause infections in
animals and man following contact with soil
Species:
Ø Microsporum gypseum
Ø Microsporum fulvum
Ø Microsporum nanum
Ø Microsporum cookei
Ø Trichophyton terrestre
Ø Trichophyton ajelloi
Are primarily parasitic on animals
Infections may be transmitted to man following
contact with animal host
Common species:
Ø Microsporum canis
Ø T. mentagrophytes var.
mentagrophytes
Ø Trichophyton equinum
Ø Trichophyton verrucosum
Ø T. mentagrophytes var. quinckeanum
Ø Microsporum canis var. distortum
Ø Microsporum equinum
Ø T. mentagrophytes var. erinacei
Are primarily parasitic on man and have only rarely been known to infect animals, presumably following contact with man
Such as:
Ø Microsporum audouinii,
Ø Trichophyton rubrum,
Ø Trichophyton schoenleinii,
Ø Trichophyton tonsurans and Trichophyton violaceum are unable to colonize animals other
than man
The only fungi that have developed a dependency on man for the maintenance and dissemination of their species
Species:
Ø Trichophyton rubrum
Ø Trichophyton tonsurans
Ø Trichophyton rubrum granular type
Ø T. mentagrophytes var. interdigitale
Ø Epidermophyton floccosum
Ø Trichophyton violaceum
Ø T. mentagrophytes var. nodulare
Ø Trichophyton concentricum
Ø Microsporum ferrugineum
Ø Trichophyton schoenleinii
Ø Trichophyton soudanense
Ø Microsporum audouinii
Classification of tinea based on areas of body involved § In cases of vesicular tinea pedis
§ Tinea capitis – head or scalp - The tops of any fresh vesicles should be removed as the fungus is often plentiful in the roof
- Three types of in vivo hair invasion are recognized: of the vesicle
1. Ectothrix invasion is characterized by the development of arthroconidia on the outside of the hair § Skin and nail specimens may be scraped directly onto special black cards
shaft. - Make it easier to see how much material has been collected and provide ideal conditions
¨ The cuticle of the hair is destroyed and infected hairs usually fluoresce a bright greenish for transportation to the laboratory
yellow color under Wood’s ultraviolet light § Skin scrapings, nail scrapings and epilated hairs should be examined using 10% KOH and Parker
¨ Common agents include M. canis, M. gypseum, T. equinum and T. verrucosum
2. Endothrix hair invasion
ink or calcofluor white mounts
¨ Characterized by the development of arthroconidia within the hair shaft only
¨ The cuticle of the hair remains intact and infected hairs do not fluoresce under Wood’s o Trichophyton schoenleinii
ultraviolet light -Anthropophilic fungus causing favus in humans
¨ All endothrix producing agents are antropophilic e.g. T. tonsurans and T. violaeum -Chronic, scarring from of tinea capitis characterized by saucer shaped crusted lesions and
3. Favus permnanent hair loss
¨ Usually caused by T. schoenleinii, produces favus-like crusts or scutula and corresponding -Invaded hairs remain intact and fluoresce a pale greenish yellow under Wood’s ultra violet light
hair loss
- Key features: Culture characteristics, microscopic morphology showing favic chandeliers and clinical
§ Tinea barbae – beard or mustache Disease
§ Tinea corporis – body “ring worm” -antler “nail head” hyphae or “favic chandelier”
- Refers to dermatophytosis of the glabrous skin
- May be caused by anthropophilic species such as T. rubrum o Basis for ID of dermatophytes
- Uusually by spread from another body site or by geophilic and zoophilic species such as M. 1. Colonial appearance
gypseum and M. canis following contact with either contaminated soil or an animal host 2. Microscopic morphology
§ Tinea cruris – inguinal area “Jock’s itch” - Macroconidia and microconidia
- Dermatophytosis of the proximal medial thighs, perineum and buttocks Ø Present or absent
- It occurs more commonly in males Ø Characteristic appearance
- Is usually due to spread of the fungus from the feet
- Thus the usual causative agents are T. rubrum, T. interdigitale and E. floccosum o Preference
§ Tinea pedis – feet “Athlete’s foot”
- Usually caused by the shedding of skin scales containing viable infectious hyphal elements (arthroconidia)
Fungus Hair Skin Nails
of the fungus Microsporum + + -
- Desquamated skin scales may remain infectious in the environment for months or years Trichophyton + + +
- Therefore, transmission may take place by indirect contact long after the infective debris has been shed
- Carpet and matting that hold skin scales make excellent vectors Epidermophyton - + +
- The toe web spaces are the major reservoir on the human body for these fungi
- It is not practical to treat infections at other sites without concomitant treatment of the toe web spaces Microscopic M. audouinii M. canis M. gypseum
- Trichophyton rubrum,T. interdigitale (Mentagrophytes var interdigitale) and Epidermophyton floccosum
§ Tinea unguium – nail “onichomycosis” Macroconidia Rare numerous

o Clinical manifestations: Spindle shaped, thick walled echinulate

§ Tinea barbae, tinea capitis, tinea mannum
- 80% - T. rubrum
Abortive, bizarre Knob like end End not pointed
- 10% - T. mentagrophytes var interdigitale
- 5% - E. floccosum
Microconidia few
§ Tinea cruris or jockey itch
- 90% in males Sessile Short pedicle clavate
Ø Unilateral lesions – T. rubrum
Ø Bilateral lesions – E. floccosum Hyphae Usually sterile Chlamydospore Racquet or antler hyphae
§ Tinea pedis or athletes foot
- Most common cause – T. mentagrophytes, T. rubrum, E. floccosum
§ Tinea unguium or onychomychosis o How to distinguish the different dermatophytes
- T. rubrum, E. floccosum, T. mentagrophytes - M. audouinii
§ Dermatophytid or Id reaction Ø When present, macroconidia may resemble M. canis but are usually longer, smoother
-A dermatophyte infection in one area elicits an allergic reaction elsewhere
and more irregularly fusiform in shape
- M. canis
o Laboratory Examination:
Ø Spindle shaped, thick-walled, verrucose macroconidia (5-15 cells) with terminal knob
1. Collection of specimen
- M. gypseum
2. Direct examination of specimen – 10% KOH wet mount
Ø Symmetrical, ellipsoidal, thin-walled, verrucose 4-6 celled macroconidia
KOH: branching hyphae or chains of arthroconidia (arthrospores)
Ectothrix: dense sheaths of spores around the hair (Microsporum)
Endothrix: (T. violaceium)
T. mentagrophytes T. schoenleinii
3. Culture - Spherical microconidia often forming - Characteristic antler “nail head” hyphae also known
- SDA with and without antibiotics dense clusters as ”Flavic chandeliers”
- Keep for 1 month at room temperature - Spiral hyphae
§ Clinical material: Skin scrapings, nail scrapings and epilated hairs - Smooth, thin-walle, clavate, multiseptate E. floccusum
§ In patients with suspected dermatophytosis of skin (tinea or ringworm) macrocondia - Smooth, thin-walled macroconidia, often produced in
T. rubrum clusters growing directly form hyphae
- Any ointments or other local applications present should first be removed with an alcowipe
- Scanty to moderate number of slender - Numerous chlamydoconidia formed in older cultures
§ Using a blunt scalpel, tweezers, or a bone curette, firmly scrape the lesion, particularly at the clavate to pyriform microcondida
advancing border and creases - Macroconidia are absent
-
SUBCUTANEOUS MYCOSES

Disease Causative organisms

Sporotrichosis Sporothix schenckii
Chromoblastomycosis Fonsecaea, Phialophora,Cladosporium etc.
Phaeohyphomycosis Cladosporium, Exophiala,Wangiella, Bipolaris, Exserohilum, Curvularia
Mycotic mycetoma Pseudaliescheria, Madurella, Acremonium, Exophiala etc
Subcutaneous zygomycosis (Entomophthoromycosis) Basisiobolus ranarum, Conidiobolus coronatus

Subcutaneous zygomycosis (Mucormycosis) Rhizopus, Mucor, Rhizomucor, Absidia, Seksenaea etc

Rhinosporidosis Rhinosporidium seeberi Rare
Lobomycosis Loboa loboi Rare

• Are chronic, localized infections of the skin and subcutaneous tissue

• Follows the traumatic implantation of the etiologic agent
• The causative fungi are all soil saprophytes of regional epidemiology whose ability to adapt to the tissue environment and elicit disease is extremely variable

SPOROTRICHOSIS “Rose Gardener’s Disease” Chromoblastomycosis

o A chronic mycotic infection of the cutaneous or subcutaneous tissues and adjacent
lymphatics
o Characterized by nodular lesions which may suppurate and ulcerate ; Lesions- non
healing ulcers
o Traumatic implantation of the fungus into the skin ; (thorns, splinters) usually to arm
hand
o Or very rarely, by inhalation into the lungs
o Secondary spread to articular surfaces, bone and muscle is not infrequent
o And the infection may also occasionally involve the central nervous system, lungs or
genitourinary tract
o Etiologic agent: Sporothrix schenckii, commonly found in soil and on decaying
vegetation
o Distribution: worldwide (tropical and temperate regions)
o Laboratory diagnosis
§ Tissue biopsy is the best specimen; Specimen: exudate from lesions
§ Direct Exam- little diagnostic value, difficult to demonstrate yeast forms
§ Tissue sections should be stained using PAS digest, Grocott’s methenamine silver
(GMS) or Gram stain
§ Look for small narrow base budding yeast cells (2-5µm)
- Note they are often present in very low numbers and may be difficult to find
- PAS and GMS stains are essential
1. Culture – SDA, BHIBA
- SDA at room temp – rapidly growing (3-5 days) mold colony
Ø Small, moist, white to cream colored colonies → membranous, wrinkled,
dark brown or black → leathery consistency
Ø Microscopic exam (LPCB) – delicate, septate, branching hyphae, conidia
are borne in bouquet-like clusters from tips of conidiophores (flowerette
arrangement)
- BHIBA at 37°C – yeast colonies are soft, cream colored to white
Ø Microscopic exam (gram stain) – oval or cigar shaped yeast cells, single
or multiple budding
o A mycotic infection of the cutaneous and subcutaneous tissues
o Characterized by the development in tissue of dematiaceous (brown-pigmented),
planate-dividing, rounded sclerotic bodies
o Infections by the traumatic implantation of fungal elements into the skin
o Are chronic, slowly progressive and localized
o Tissue proliferation usually occurs around the area of inoculation producing crusted,
verrucose, wart-like lesions (cauliflower-like)
o Distribution: World-wide but more common in bare footed populations living in tropical
regions
o Etiologic agents
o Various dematiaceous hyphomycetes associated with decaying vegetation or soil,
especially
§ Phialophora verrucosa, Fonsecaea pedrosoi, F. compacta and Cladosporium
carrionii
o Laboratory diagnosis
§ Specimen – scrapings from crusted areas and/or biopsy
§ KOH wet mount – sclerotic bodies which appear as rounded and brown structures
(copper pennies)
§ Cultures of the aetiologic agents of chromoblastomycosis are typically olivaceous-
black with a suede-like surface
§ The dematiaceous hyphomycetes involved are well recognized as environmental
fungi
- Therefor a positive culture from a nonsterile specimen, such as sputum or
skin, needs to be supported by clinical history and direct microscopic
evidence in order to be considered significant
§ Generic criteria are based on the type and ontogeny of conidial production
- Cladosporium type of conidiophore – blastoconidia in chains
- Phialophora type of conidiophores – phialides with conidia
- Acrotheca type of conidiophores – annelides, sympodulae
MYCETOMA
o A mycotic infection of humans and animals caused by a number of different
fungi and actinomycetes characterized by:
§ Draining sinuses,
§ Granules and
§ Tumefaction (swelling)
o Usually involves the cutaneous and subcutaneous tissue, fascia and bone of
the foot or hand
o Usually result from traumatic implantation
o Sinuses discharge serosanguinous fluid containing the granules
§ Granules are the hallmark of mycetoma
§ Vary in size, colour and degree of hardness, depending on the aetiologic
species, and
o Distribution:
§ World-wide but most common in bare-footed populations living in
tropical or subtropical regions
o Aetiologic agents: Actinomycetoma vs. Eumycetoma (Madura Foot)
§ Actinomycotic mycetoma: Nocardia, Actinomadura and Streptomyces
§ Eumycotic mycetoma: Madurella, Acremonium, Pseudoallescheria,
Exophiala, Leptosphaeria, Curvularia, Fusarium, Aspergillus, etc
Lab diagnosis
§ Clinical material: Tissue biopsy or excised sinus, serosanguinous fluid containing the granules which vary in size, colour and
degree of hardness, depending on the aetiologic species
§ Serosanguinous fluid containing the granules should be examined using either 10% KOH and Parker ink or calcofluor white
mounts, and tissue specimens should be stained using H&E, PAS digest, and Grocott’s methenamine silver (GMS)
§ The presence of white to yellow or black pigmented grains, from a patient with supporting clinical symptoms should be
considered significant
§ Biopsy and evidence of tissue invasion is of particular importance
Madura Foot, Maduromycosis:
Pseudoallescheria boydii: most common cause in the USA; greyish mold colonies producing conidia and ascospores
ENDEMIC MYCOSES
o Usual portal of entry – LUNGS (conidia) à yeast cells or sperules (Coccidiodes)
o Endemic areas = high infection rates; 90% among immunocompetent à asymptomatic/ self limited
o Organisms with organ predilection:
-Histoplasma capsulatum – RES -Blastomyces dermatitidis – cutaneous, muco
cutaneous tissues, bones

SYSTEMIC MYCOSES

Blastomycosis (Gilchrist’s Disease) Paracoccodioidomycosis

Coccidioidomycosis Histoplasmosis
(South American Blastomycosis)
Etiology Coccidioidis immitis Histoplasma capsulatum Blastomyces dermatitidis Paracoccidioides brasiliensis
Habitat Soil in south west USA Soil, bat and avian habitat Soil
Portal of Respiratory tract (inhalation)
Respiratory tract and inhalation of arthroconidia
entry
Lungs, skin, bones, soft tissue Asymptomatic, mucous membrane, nose,
60% no ssx , RT 95% no ssx, RES
Organs mouth, conjuctiva
RT, lungs, meninges, bones, visceral organs Liver, spleen, heart, meninges
Sputum, urine Sputum, pus, exudates Sputum, tissue scrapings
Specimens
Exudate- cutaneous ; Blood, CSF Scrapings, BM, buffy coat cells
KOH: thick walled non budding IC spherules Wright: blood, BM; small ovoid yeast cells w/in 10% KOH wet mouth: large, spherical thick- Large, round to oval multiple budding yeasts
Direct Exam
with endosores macrophages walled cell with single bud, w/ broad base cells- mariner’s wheel or steering wheel
culture Dimorphic, mold colony at room temp. Dimorphic, mold colony at room temp.
Mold colony Cottony, white to gray, fluffy, cobweb like Fluffy, white to tan, slow growing Slow growing cottony, white tan “prickly state” Grow very slowly white to brown
Barrel shaped arthroconidia, alternate Large thick walled yeast cells, single bud Yeasts with multiple narrow necked buds
Tuberculate macroconidia; microconidia
Microscopic arthroconidia, racquet hyphae easily attached by broad base “steering wheel”
delicate round to pyriform, smooth
aerosolized
Yeast colony @ 37 deg Celsius, slow growth
BHIBA
Smooth, wrinkled, tan White, cerebriform, mucoid colony
Microscopic Tissue cultures produce spherules Budding ellipsoidal yeasts cell
Serology Precipitin test, CFT agglutination test Immunodiffusion, CFT Precipitin CFT
Skin test Coccidiodin, Spherulin Histoplasmin Blastomycin Paracoccidiodin

OPPOTUNISTIC MYCOSES

Predisposing Factors: Immunologic disorders, diabetes mellitus, corticosteroids

Mechanisms of host resistance to fungal infections
1. Neutrophil
Fungicidal activity: kills yeast cells of C. albicans & hyphae of C. albicans, A. fumigatus and Rhizopus ; ineffective in killing the tissue forms of primary pathogens
2. Intact CMI and activated Macrophages crucial for adequate defense against C. immitis and H. capsulatum, B. dermatitidis, P. brasiliensis, C. neoformans
Antibodies and other humoral factors: not protective against primary or opportunistic fungi
CANDIDIASIS CRYPTOCOCCOSIS

Etiologic agent: Candida albicans Chronic, subacute to acute pulmonary, systemic or meningitic disease, initiated by inhalation
Non albicans: C. glabrata, C. tropicalis, C. krusei, C. parapsilosis, C. guilliermondii, C. pseudotropicalis o Primary pulmonary infections usually subclinical
o Predilection for CNS
Morphology and Physiology:
o Produce yeast cells, pseudohyphae, true hyphae Etiologic agent: Cryptococcus neoformans
o When in normal flora= seen as budding yeast cells
o During tissue invasion= hyphal form Determinants of pathogenicity:
o Normal serum- stimulant for conversion of yeast cells to hyphae o Capsule: inhibits phagocytosis of yeats cells, inhibits production of capsular antibodies
o Incubation in serum at 37 deg Celsius for 90 mins à Germ Tube formation o Diphenol oxidase production: assoc. w/ virulence, pathogenicity not clear

Determinants of Pathogenicity: Cultural characteristic:

o Hyphal production o Uniformly spherical, budding, capsulated yeast cell in tissue and culture
o Resistance to phagocytic killing o Yeast cells (5-10 u) with single or multiple budding
o Endotoxin like toxicity o Capsule: hallmark of C. neoformans (2-3 x its width)
o Adherence and penetration of host cells o Serotypes based on capsulat antigen; A (most common), B, C, D
Adheres to plastic surfaces- catheters and prosthetic devices
Diagnostic tests
Clinical classifications of Candidiasis o Culture: rapid growth on SDA
o Cutaneous and Subcutaneous Candidiasis Bird seed (niger seed) media- brown pigment of colonies is
Thrush, stomatitis, intertriginous candidiasis, onychomycosis diagnostic
o Systemic Candidiasis
Bronchopulmonary candidiasis, meningitis, pyelonephritis, myocarditis, cystitis Serologic tests:
o Cryptococcal polysaccharide capsular antigen can be performed on CSF and serum
Cutaneous Candidiasis: including intertrigo, diaper candidasis, paronychia and onychomycosis
Lesions consist of a moist, macular, erythematous rash with typical satellite lesions present on the surrounding healthy skin
Diaper candidiasis is common in infants under unhygienic conditions of chronic moisture and local skin maceration associated with ammonitic
irritation due to irregularly changed unclean diapers
Once again characteristic eruthematous lesions with erosions and satellite pustules are produced, with prominent involvement of the skin folds
ASPERGILLOSIS SYSTEMIC ZYGOMYCOSIS (MUCORMYCOSIS)

Aspergillus sp. o Acute and rapidly developing infection of debilitated patients

o Isolated from vegetables, esp nuts and grains; from decaying matter, soil and air o Involves rhino-facial-cranial areas, lungs, GIT, skin, less common in others
o 200 species but 4 commonly cause disease: o Predilection for invading vessels of the arterial system, causing embolization à necrosis of
A.fumigatus -most common; A. niger, A. flavus, A.terreus surrounding tissue
o 2nd most frequent cause of opportunistic fungal infection
Morphology and cultural characteristics
Morphology and cultural characteristics: o Grow rapidly
o Rapid growing fungus o Cottony / fluffy mycelia- often fills the dish
Aspergillus fumigatus Aspergillus flavus Aspergillus niger o ID by microscopic morph:
Gross: fluffy to granular, blue green to Gross: yellow Gross: black -broad, non-septate hyphae, often twisted and
grey green green colony colony ribbon-like
Mx: uniserate heads with Mx: uniserate to Mx: biserate o Asexual reproduction- sporangia
phialides covering upper ½ biserate, with phialides o ID of 3 common Zygomycetes based on presence or absence of rhizoids and position of the
to 2/3 of the vesicle phialides covering the rhizoids in relation to the sporangiophore
Thermotolerant: withstand covering entire surface of vesicle entire surface of the vesicles § Mucor- NO rhizoids
temp to 45 deg Celsius § Rhizopus- rhizoids at point where stolons arise
§ Absidia- presence of rhizoids which originate between sporangiophores
o Microscopic:
Clinical infection: § sporangium – (sac like fruiting structure) contains sporangiospores
o Inhalation of aspergillus spores/mycelial fragments: § supporting structure is the sporangiophore connected to each other by stolons,
-Disposed by the body w/o injury to the host which attach at contact points where root-like structures (rhizoids) anchor the
-May elicit an immediate hypersensitivity response w/o invasion of the body organism to the agar surface
-Grows in the bronchial tree and caused infection, allergic bronchopulmonary aspergillosis Clinical infection:
o Predisposing factors: o Epidemiology: persons at risk: patients with acidosis; leukemia, immunodefeciencies
-Structural abnormalities in the lung o Pathogenesis:
-severely impaired resistance to infection àSporangiospores germinate, and thrive in nasal, oropharyngeal or respiratory mucosa of
-therapy used in combating above disease immunocompromised patients
à Hyphae invade the lumen and walls of blood vessels causing thrombosis leading to infarction
and necrosis
Other Opportunistic mycoses: Cultural characteristic: Mx:
Branched conidiophore (brush like) , 20-30
PENICILLOSIS- pulmonary and systemic infection Colony: rapid growth, colont at first is white à characteristic for spp. branches (metulae), phialoconidia- hyaline to dark
smooth to rough
Etiologic agent: PENICILLUM Most commonly isolated spp. Has blue green to yellow green spores