Mycology - study of fungi Yeast: unicellular Molds: filamentous and multicellular

Mykes (Greek) Two Forms: Blastoconidia (budding) - Asexual Mycelium - masses of hyphae
Yeast and Ascospores or Basidiospores - Sexual Hyphae - tubelike projections
Father of Mycology:
Raymond Sabouraud Mold/Moulds
Fungal Morphology: Sexual Reprodcution:
Cell Membrane: bilayered; ergosterol Perfect Fungi
General Properties of fungi: Cell Wall: chitin; potent to humans
Eukaryotic - cell bound Capsule: polysaccahride coating; C. neoformans (India Ink) Asexual Production:
Ergosterols/Chitin (cell wall) - 80S ribosomes Imperfect Fungi
Chemoheterotrophs: req. organic compounds
Lack Chlorophyll
Osmiotrophic
Saprophytes: eat dead matter
Require Water and Oxygen
Asexual and Sexual Reproduction

3 Types of Hyphae: Pseudohyphae: contain “areas of constriction” between adjacent cells

a. Coenocytic = No septate → Hyphae of Zygomycetes
b. Dark and Pigmented Septate → Hyphae of Dematiaceous Fungi Spores: reproductive structure of fungi
c. Septate, Non-Pigmented → Hyaline Molds

Taxonomy Grouping: Germ Tube: No constriction is observed at the point of attachment

Zygomycota: sparsely septate (coenocytic) ; Asexual
(sporangiospores); Sexual (zygospores)
= Rhizopus, Mucor, Rhizomucor, Absidia, Cunninghamella,
Saksenae
Ascomycota: Asexual (conidia); Sexual (ascospores); true septate
hyphae; Teleomorph and Anamorph
=Ajellomyces, Arthroderma
Taxonomy Grouping:
Basidiomycota: Sexual (basidiospores)→Basidia
=Cryptococcus neoformans, Filobasidiella
Deuteromycota: lack sexual phase; Asexual (conidia)
= Candida, Aspergillus, Sporothrix, Coccidiodes, Trichophyton, etc.
Topographic Grouping of Fungi
A. Superficial or Cutaneous Mycoses: skin, hair; non-invasive; patchy scaling, eczema eruptions;
classified according to the area of the body that is involved; infects Keratinized tissues
=Dermatophytes, Agents of Tinea
B. Subcutaneous Mycoses: deeper layers of skin; muscle tissue w/o dissemination, Man (accidental
host); tissue rx or granule
=Cladosporium, Fonsaceae, Exophiala, Phialophora, Mycetoma agents
C. Systemic Mycoses: deep tissues and organ systems; dimorphic fungi
=Histoplasma capsulatum, Blastomyces dermatitidis, Sporothrix schenckii, Coccidiodes
immitis, Paracoccidiodes brasiliensis, Penicillium manrneffei
D. Opportunistic Mycoses: immunocompromised patient, underlying dse.,
=Aspergillus, Fusarium, Geotrichum candidum, Acremonium, Penicillium, Paecilomyces,
Scopulariopsis

Sexual Reproduction: meiosis → merging of cells and nuclear fusion, (BAZO-S)

Ascospores: saclike ascus (8)
Zygospores: fusion of cells from 2 identical cells, same hypha
Oospores: fusion of cell from 2 separate non-identical hypha
Basidiospores: club shaped basidium
**Teleomorph = fruiting body
Asexual Reproduction: mitosis only
Conidiogenesis: conidia formation - blastic or thallic
a. Blastic Conidiogenesis: parent cell enlarges
→septum→enlarged splits off→daughter cells
b. Thallic Conidiogenesis: septum→new growth beyond
septum→daughter cells
Asexual Reproduction: from conidia
Conidia: multiply in long chains or clusters by specialized
vegetative hyphae known as conidiospores.
Asexual Reproduction: from thallus (BACS-A)
a. Blastoconidia: daughter cells buds off mother cell
b. Chlamydoconidia: thick-walled resistant resting spores
produced by rounding up and enlargement of the terminal
hyphal cells
c. Arthroconidia: simple fragmentation - cylinder-shaped or
caske-shaped spores
d. Sporangiospores: contained in sporangia or sacs, aseptate
hyphae; sporulation→sporangial wall ruptures

Arthroconidia
 LABORATORY METHODS IN MYCOLOGY
Skin: 70% alcohol to remove dirt, oil and surface saprophytes
Direct Examination of Specimens:
a. Direct Microscopre Examination: required on any material
- spores, hyphae, mycelial elements, budding yeast, mycotic
granules.
b. Wet Mount Prep: good for yeast
c. KOH prep: skin scrapings, hair, nails, sputum, vaginal spx.,
etc.; clears the spx's tissue cells
Media Used for Isolation:
1. SDA - classic medium, recommended for most studies
2. SDA w/ Chloramphenicol: inhibits bacteria
3. Mycosel Agar: chloramphenicol (bacteria); cycloheximide
(saprophytic fungi and some yeasts (including C.
neoformans)
4. BHI slant: more enriched than SAb-Dex; recover H.
capsulatum
5. PDA/ Cornmeal Agar: slide cultures; induce spore
formation
Considerations and Requirements:
1. Cultures are handled in Class II VSC
2. Gas Flames - suitable for decontamination of loop used for
transfer of yeast cultures.
3. Plates are preferred
4. 40mL of agar (7-8mm) - used to prevent drying
(dehydration)
Staines Used in Mycology
Lactophenol Cottonblue: popular; chitin in CW
Gomori Methenamine Silver Stain: polysasscharide; Fungi (pink-red), Nuclei
(blue)
Gridley stain: outline (black - silver precipitating in CW), Internal structures (deep
rose - black), Bg (light green)
Mayer Mucicarmine: Hyphae & yeast (dark blue or rose), Tissue (deep blue), Bg
(yellow)
India Ink: capsule of C. neoformans (deep rose)
FAS: simple, sensitive, and specific
Pap Stain: dimorphic fungi
Gram-stain: most fungi are gram positive
Mod. Acid-Fast: diff. Nocardia from other aerobic Actinomyces
Giemsa: blood and BM
Laboratory Considerations:
Respiratory Spx: 0.5 mL of spx =sputum, BAL, bronchial washing
CSF: filter (0.45um); >1mL - centrifuged, processed ASAP, store in 30°C
Blood: Accurate method of determining fungal etiology; dimorphic fungi; 30°C -
incu" 21 days
Hair, Skin, and Nail Scraping: dermatophyte; not refrigerated
Urine: processed ASAP; centrifuged; media needs antibacterial agents
Tissue, BM, and sterile fluids: proceessed ASAP, at least 1mL for inoculation;
incu:21 days in 30°C
5. Incu at RT at 30°C for 21-30 days
6. Cultures examined for 3x weekly during incubation
Biochemical Tests:
Assimilation: ability of an isolate to utilize carbohydrate
as the source of carbon
Fermentation Tests: Anaerobic utilization of
carbohydrates with the production of gas (for ID of
Candida spp.)
Growth Enhancement:
Inositol and Thimine: Trichophyton
Peanut Oil: Malassezia
Phenol Oxidase: C. neoformans = BROWN PIGMENT
Urease Test: Cryptococcus (+) vs Candida (-)
Urease (+) = Rhodotorula, Trichosporon beigelli,
Trichophyton mentagrophytes (alkalination of medium)
 SUPERFICIAL AND CUTANEOUS MYCOSES Pedraia hortae Malassezia furfur (Pityrossporum
= Black piedra (common in scalp hair) orbiculare)
hard, dark brown to black gritty nodules = Tinea versicolor or Ptyriasis versicolor
Superficial Mycoses - affect cornified layers (stratum made up of asci containing eight patchy lesions or scaling of varying
corneum) of the epidermis; does not activate tissue ascospores pigmentation
response or inflx rx; cosmetic effects Hair shafts→ 10-20% KOH Fawn-colored lesions → chest, trunk, or
Thick-walled rhomboid cells containing abdomen
ascospores are seen. common endogenous skin colonizer
Trichosporon beigelii SDA on RT Yellow fluorescence on Wood's lamp
= White piedra → facial, genital hair Dematiaceous septate hyphae Spaghetti and Meatballs on KOH
characterized by a soft mycelia mat Tx: removal of infected hairs and Tx: 1% Selenium Sulfide
Opportunistic system pathogen fungicides
occurs most often in the immunocompromised host
Dx: hyphal elements within the shaft nodule and
budding
blastoconidia and arthroconidia in culture
Colonies are cream colored and yeast looking but
eventually becomes wrinkled as they
mature
Does not ferments carbohydrates and potassium
nitrate, assimilates glucose, galactose, sucrose,
maltose and lactose

Phaeoannellomyces werneckii / Exophiala werneckii

= Tinea nigra→ black non-scaly macules on
palms and soles
Dx: skin scrapings placed in 10-20% KOH
Septate hyphae and budding; shiny, moist, yeast-
looking colonies; brownish→olive to greenish black
Budding blastoconida
Annelloconidia = older colonies
Agents of Dermatophytes (Cutaneous Mycoses): Laboratory Diagnosis:
Microsporon Initial Dx: Potassium Hydroxide - mounted
Trichophyton skin scrapings
Epidermophyton KOH
Hair Perforation Test: T.rubrum (surface
erosion); T. mentagrophytes
Clinical Infections: (perpendicular pegs)
Tinea - "body site""ringworm"; local inoculation in Urease Test: T. mentagrophytes (+)
skin; enlarged with time; growing edge; Thiamine Requirement: most useful
nutritional test for dermatophytes
Growth on Rice Grains: M. canis (+);
Epidermophyton flocussom: M. audouinii (NG)
Anthromorphic - skin and nails
Colony; yellow-green, no microconidia
"Beaver's tail" Microsporum gypseum:
Notorious for devp. pleomorphic tufts Rapid growing geophilic
Skin and hair
Macroconidia and microconidia - powdery,
granular appearance
Tan to buff conidial masses
Fusiform mod. thick walled conidia
Microsporum audouinii:
Slow growing anthrophilic
Gray-patch worm in children
Skin, Hair
Colony: cotton-white form, no pigment on
reverse
Some form chlaymydoconidium-like
swelling on terminal hyphae
Trichophyton mentagrophytes:
Most isolated
Skin, Hair, Nails
powdery appearane, abundant microconidia
(clusters
Trichophyton rubrum:
Most common spp isolated for ringworm
Slow growing anthrophilic (human)
dermatophyte
Skin and Nails
Hyaline →pink-deep burgundy wine colored
pigment on reverse side
3-8 macroconidia; clavate/peg-shaped
microconidia
Trichophyton tonsurans:
Tinea capitis in children
Skin, Hair, Nails
Rust colored colonies on reverse SDA
Balloon-forms microconidia
Reqs Thiamine
Microsporum canis:
Rapid growing zoophilic
most commonly isolated Microsporum spp.
Macroconidia (spindle-shaped) and
microconidia
Lemon-yellow on PDA (reverse side)
 SUBCUTANEOUS AND SYSTEMIC MYCOSES Subcutaneous Mycoses:
Chromoblastomycosis
Systemic Mycoses:
Blastomyces dermatitidis
Mycetoma Coccidiodes immitis
Subcutaneous mycoses - traumatic skin Phaehypomycosis Histoplasma capsulatum
puncture→thorns and vegetation; chronic and localized Sporothrichosis Paracoccidiodes brasiliensis
infections; soil saprophytes; "Inoculation Mycoses" Penicillium marneffei
Chromoblastomycosis: Mycetoma:
Dematiaceous fungi- dark, slow-growing Chronic granulomatous infection; tumorlike deformoties→ abscess, draining sinuses
Skin puncture or trauma Pus: yellow, red, white or black granules
Chronic, non-healing, hard, warty, tumor-like Occupation infx → seen on feet
lesions→ feet and lower legs
Sclerotic bodies/Medlar bodies = "copper-colored, 1. Exophiala jeanselme
septate cells" 2. Pseudollescheria boydii
1. Fonsecaea pedrosoi 3. Acremonium
2. Fonsecaea compactum 4. Curvularia
3. Phialophora verrucosa 5. Madurella
4. Cladosporium carrioni
Sporotrichosis:
Phaeohypomycosis: Sporothrix schenkii
Infx of any dematiaceous fungi; darkly pigmented, septate Dimorphic fungi
hypahe Skin trauma→ thorny plants esp rose (Rose Gardner's Disease)
May lead to systemic infection Local and regional lymphadenopathy. may disseminate
Yellow-brown (melanin pigmentation), w/ or w/o budding Inhalation
G on SDA at 25°C; small, cream-colored wrinkled, leathery
1. Bipolaris colonies→black
2. Phialophora richardsiae BHI: Yeast at 37°C
3. Wangiella dermatitidis May appear intracellularly to PMNs

"Flowerette
Appearance"

SUBCUTANEOUS AND SYSTEMIC MYCOSES
Features of Systemic Mycoses:
Human: dead end
Cause dse in healthy host
MOT: Inhalation
Host Response: Cell-mediated (granulomatous)
Histoplasma capsulatum:
Histoplasmosis. Ohio Valley Fever, Darling's Dse
Chicken, pigeon dropping or bat guano
Inhalation of spores
Bird roosts, chicken houses, barns, and bat caves
RTI→fatal pulmonary disease
Chronic form may resemble tuberculosis
Midwestern and southern US; endemic in Mississippi
and Ohio River Valleys
Blastomycosis dermatitidis: Penicillium marneffei:
Blastmycosis; North American Blastomycosis / Penicilliopsis
Gilchrist's Disease Acute infx of lungs, bone
Chronic infx: lesions or ulcerations→lungs, skin, and marrow
bone SEA, China, India, and Hong
North America, Mississippi, Missouri, Ohio River Valleys Kong
Round-oval, thick-walled, broad-based, single budding Px w/ AIDS in SEA (Thailand,
Mycelial phase: lollipop (hyphae) Taiwan, and India)
Acne-like skin papules on
face, trunk, and extremities
Paracoccidiodes brasiliensis
South American Blastomycosis
Granulomatous dse→lungs, lymphatics, skin, mucus
membrane
Brazil, Venezuela, Colombia
Inhalation; may disseminate into the liver and spleen
Multiple budding yeast "Mariner's Wheel"
Mycelial: Intercalary and terminal chlamydoconidia
OPPORTUNISTIC MYCOSES YeastOpportunistic
and Yeast Infections
Opportunistic Mycoses:
1. Candida
2. Cryptococcus
3. Zygomycosis
4. Pneumocystis jiroveci
Cryptococcus
C. neoformans and C. gatti
Systemic infx in healthy and
immunocompromised
High mortality rate; cryptococcal meningitis - high
incidence of HIV-infected px
Occurs in lungs - inhalation
Disseminated→CNS
Accompanied by Pneumocystis or CMV
India ink preparation for capsules
Rapid Urease (+)
Gram stain (CSF): starburst appearance
Zygomycosis
Invasive fungal infx
Mucormycosis or phycomycosis
Rhizopus spp: most commobly isolated
Clinical presentation: sinus infx or orbital
involvement; pulmonary dse.
Candida
Most frequently encountered opportunistic fungal infx
Sputum, urine,vagina or stool (not Candida albicans: most frequent agent
pathogenic) Human colonizer and mucous membrane
Pathogenic: in sterile sites Normal flora; skin, mouth, vagina, stool
Cornmeal Agar Environment: leaves, flowers, water, and soil
Asexual: Blastoconidia Candidiasis
Sexual: Ascospores or Basidiospores Hematogenous spread→disseminated infections
Cutaneous Infections - erythematous lesions, creamy, white
exudates or scaling (groin, bet. fingers and toes, axilla, under
female breasts)
= Chronic Cutaneous Candidiasis: px w/ defective immunity
Oral Candidiasis - oral thrush; fissuring at the corners of mouth;
common initial infx of px w/ HIV →immune failure marker
GI Candidiasis - esophagitis, px w/ HIV (ART Thx)
Vaginal Candidiasis - vaginal thrush; vaginal burning and itching,
dyspareurnia; curd-like discharge
Invasive Candidiasis - hematogenous spread
Candida
Most frequently encountered opportunistic fungal infx
1. Absidia Candida albicans: most frequent agent
2. Cunninghamella Human colonizer and mucous membrane
3. Mucor Normal flora; skin, mouth, vagina, stool
4. Rhizomucor Environment: leaves, flowers, water, and soil