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Mycology Terms

Moulds: Multicellular Fungi Yeasts: Single-cell Fungi Mycosis: Fungal Infection Systemic Mycosis: Multi-organ infection caused by fungi Opportunistic mycosis: Fungal disease that occur primarily in immunocompromised patients • Dimorphic fungi: Fungi that show both a non-mould and mould phase • Saprobe: Organism capable of living on decaying organic Material • • • • •

The Characteristics of Fungi
• Body form – unicellular – filamentous (tube-like strands called hypha (singular) or hyphae (plural) – mycelium = aggregate of hyphae – sclerotium = hardened mass of mycelium that generally serves as an overwintering stage. – multicellular, such as mycelial cords, rhizomorphs, and fruit bodies (mushrooms)

The Characteristics of Fungi
• Heterotrophy - 'other food' – Saprophytes or saprobes - feed on dead tissues or organic waste (decomposers) – Symbionts - mutually beneficial relationship between a fungus and another organism – Parasites - feeding on living tissue of a host. • Parasites that cause disease are called pathogens.

Fungi as Saprobes and Decomposers .

Fungi as Symbionts (Mutualism) .

Fungi as Parasites & Pathogens .

Heterotrophic by Absorption • • • • Fungi get carbon from organic sources Hyphal tips release enzymes Enzymatic breakdown of substrate Products diffuse back into hyphae Nucleus hangs back and “directs” Product diffuses back into hypha and is used .

branching filaments that come together to form the mycelium – Septate hyphae – with ceparation or cross walls – Aseptate or Sparsely septate – Pseudohyphae – chain of cells formed by budding • Vegetative Hyphae – functions as food absorption and are the portion that extends below the agar surface • Aerial Hyphae – extends above the agar forming conidia .Fungal Structures • Hyphae – long.

Conidia Types of Conidia • Arthroconidia – resulting from the fragmentation of hyphae into • Conidial morphology individual cells is important in fungal • Balstoconidia – conidia formed identification as the result of budding • Microconidia – small • Chlamydoconidia – terminal cells in the hyphae enlarge and • Macroconidia – large have thick walls • Poroconidia – formed by being pushed through a small pore • Phialoconidia – tube shaped that can be branched • Annelloconidia – vase shaped conidia .

Arthroconidia Blastoconidia Chlamydoconidia Poroconidia .

Phialoconidia Annelloconidia .

5-3 Key Haploid (n) Heterokaryotic Diploid (2n) Spore-producing structures Spores Mycelium ASEXUAL REPRODUCTION PLASMOGAMY Heterokaryotic stage KARYOGAMY SEXUAL REPRODUCTION Zygote GERMINATION GERMINATION MEIOSIS Spores .Figure 31.

they coexist in the mycelium. with the exception of transient diploid stages formed during the sexual life cycles • Sexual reproduction requires the fusion of hyphae from different mating types • Fungi use sexual signaling molecules called pheromones to communicate their mating type • Plasmogamy is the union of cytoplasm from two parent mycelia • In most fungi. the haploid nuclei from each parent do not fuse right away. such a mycelium is said to be dikaryotic . the haploid nuclei pair off two to a cell. called a heterokaryon • In some fungi.SEXUAL REPRODUCTION • Fungal nuclei are normally haploid.

producing diploid cells • The diploid phase is short-lived and undergoes meiosis. or even centuries may pass before the occurrence of karyogamy. many fungi can reproduce asexually Molds produce haploid spores by mitosis and form visible mycelia .• Hours. the haploid nuclei fuse. days. nuclear fusion • During karyogamy. producing haploid spores • The paired processes of karyogamy and meiosis produce genetic variation ASEXUAL REPRODUCTION In addition to sexual reproduction.

silver nitrate outlines fungi in black due to the silver precipitating on the fungi cell wall. • Gomori Methenamine Silver Stain .FUNGAL SPECIES IDENTIFICATION METHODS Mycology: Lab Methods Stains used: • Lactophenol Cotton Blue . .very popular for quick evaluation of fungal structures. stains chitin in cell walls of fungi. Fungi stain pinkred with blue nuclei. • Periodic Acid .Schiff Stain (PAS) .stains polysaccharide in the cell wall of fungi.

extremely specific method of detecting fungi in tissues or fluids.• Gridley Stain . • Mayer Mucicarmine Stain . • Papanicolaou Stain .will stain capsules of Cryptococcus neoformans deep rose. Works well on sputum smears. • Fluorescent Antibody Stain . Tissues stain deep blue and background is yellow. • India Ink .Hyphae and yeast stain dark blue or rose.used for blood and bone marrow specimens.good for initial differentiation of dimorphic fungi. • Giemsa Stain . but Actinomyces and Nocardia are gram variable.used to differentiate the acid-fast Nocardia from other aerobic Actinomyces.demonstrates the capsule of Cryptococcus neoformans in CSF specimens. . • Acid-Fast Stain . • Gram Stain .generally fungi are gram positive.

• Sabouraud's dextrose agar with chloramphenicol (inhibits bacterial growth). neoformans). Preliminary identification is based on differential growth patterns on various media. recommended for most studies. and cycloheximide to inhibit saprophytic fungi and some yeasts (including C. – Easier storage. • The agar in a tube is inoculated in a straight line. Media: • Sabouraud's dextrose agar . . Bacteria-like fungi (such as Actinomycetes) are inhibited by chloramphenicol. – Less chance for dehydration. • Mycosel agar . – Aspergillus and Scopulariopsis are opportunistic pathogens. Cycloheximide will prevent their growth.Fungal Culturing: • Tubed media is used rather than plated media. – Less chance for spore release into the environment.a commercially produced agar containing chloramphenicol to inhibit bacterial growth.classic medium.

Cryptococcus neoformans will produce melanin resulting in black colonies (must protect media from light). • KT Medium & Kelley Agar .used in slide cultures to induce spore formation.• Brain heart infusion slant (BHI) . • Modified Converse Liquid Medium (Levine's) .more enriched than Sab-Dex. . • Potato-dextrose agar (PDA) and Corn-meal agar . capsulatum.used to isolate Cryptococcus neoformans from contaminated cultures.used to convert dimorphic fungus Blastomyces dermatitidis from mycelial to yeast form.used to promote spherule production by Coccidioides immitis. Special applications agar: • Caffeic Acid Agar . • Birdseed Agar . For recovery of H. which aids in identification.

– Nocardia sp. • Atmosphere . – Any fungus capable of growing at 37 C. and some dimorphic organisms grow best at 37 degrees C. • Routine cultures should be examined every other day. .room temperature (25-30 C. 10 weeks are recommended if Histoplasma capsulatum is suspected. Saprophytes are fast growers (several days).) for most fungi. • Time . should be considered potentially pathogenic. Generally cultures are held at least 4 weeks.True fungi are aerobic but there are a few anaerobes among the bacteria-like fungi. – Paracoccidioides brasiliensis may require 4-5 weeks.Fungal growth requirements: • Temperature .Some yeasts grow overnight.

creamy). velvety. powdery. or on the bottom of the colony (reverse pigmentation). .cottony or wooly (floccose). • Pigmentation . on its sporulating apparatus. • Mycelium – Vegetative mycelium – provides nutrition. on the agar. or waxy.Colony Morphology (macroscopic features): • Surface topography . Color may be on fungus itself.some fungal colonies may be free growing. – Aerial mycelium – reproductive.Fungi may be colorless or brightly colored. others may grow in a restricted manner. silky. • Surface texture . chalky. granular. covering the entire surface of agar. glabrous (smooth.

wet mount • Slide Culture . • Cellophane Tape Preparation Biochemical studies . Observe every 48 hours for 14 days. Gas is detected by appearance of bubbles trapped in the fermentation tube.Microscopic evaluation: • Teased Preparation .used to ID yeast and yeast-like organisms. . Bromcresol purple is the indicator. • Carbohydrate fermentation – Growth and utilization of a carbohydrate under anaerobic conditions as determined by acid and gas production. – Specimen is inoculated beneath the broth. Acid production turns purple to yellow.gives undisturbed microscopic morphology.

• Carbohydrate assimilation – Determines ability to utilize a carbohydrate as sole source of carbon. – Tubes unchanged (as determined by comparing to a blank tube) after 10 days are negative. . – Bromcresol purple indicator turns from purple to yellow. Bromthymol blue is the indicator (blue to yellow is positive). • Nitrogen assimilation – Utilizes 3 tubes with differing sources of nitrogen.

Growth on specific agars: • Christensen's urea agar - Urea is hydrolyzed by some yeast to form ammonia (pH increases) which turns media from yellow to dark pink. • Caffeic acid medium (must protect media from light) Production of melanin by Cryptococcus neoformans resulting in black colonies. Other tests: • Germ tube - Candidia albicans & Candidia stellatoidea produce germ tubes when incubated in a protein medium. • Demonstration of chlamydospores - Yeast is inoculated by jabbing appropriate agar (Cornmeal with tween 80) and observed every 24 hours for 3 days for chlamydospore production.

Diagnosis of Mycotic Infections
• Diagnosis and identification require microscopic examination of stained specimens, culturing in selective and enriched media and specific biochemical and serological tests

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BODY SITES AND POSSIBLE FUNGAL PATHOGENS
• Blood – Candida sp, Blastomyces dermatitidis, Histoplasma capsulatum, Cryptococcus neoformans • CSF – Cryptococcus neoformans, Candida sp. Histopasma capsulatum, and Coccidioides immitis • Hair – Microsporum and Trichophyton • Nails – Aspergillus, Epidermophyton, and Trichophyton • Skin – Candida, Microsporum, Trichophyton, Rhizopus, Penicillium, Histoplasma capsulatum, Blatomyces dermatitidis, and Coccidioides immitis • Throat – Candida albicans, Geotrichium candidum • Urine – Candida albicans, Candida glabarata • GUT – Candida albicans

dust. and normal flora • Humans are relatively resistant • Fungi are relatively nonpathogenic • Of the 100. fomites. only 300 have been linked to disease in animals • Fungi are the most common plant pathogens • Human mycoses are caused by true fungal pathogens and opportunistic pathogens 26 .Fungi as Infectious Agents • Molds and yeasts are widely distributed in air.000 fungal species.

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• True or primary fungal pathogen can invade and grow in a healthy. noncompromised host • Most striking adaptation to survival and growth in the human host is the ability to switch from hyphal cells to yeast cells • Thermal dimorphism – grow as molds at 30°C and as yeasts at 37°C 28 .

THERMAL DIMORPHISM 29 .

host defenses must be impaired • Vary from superficial and colonization to potentially fatal systemic disease • An emerging medical concern.Emerging Fungal Pathogens • Opportunistic fungal pathogen has little or no virulence. account for 10% of all nosocomial infections • Dermatophytes may be undergoing transformation into true pathogens 30 .

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phagocytosis. capsules and adhesion factors. inhaled spores – Subcutaneous – inoculated skin. inflammatory stimulants • Antifungal defenses are the integrity of the barriers and respiratory cilia • Most important defenses are cell-mediated immunity.Pathogenesis of the Fungi • Portal of entry – Primary mycoses – respiratory portal. trauma – Cutaneous and superficial – contamination of skin surface • Virulence factors – thermal dimorphism. toxin-like substances. and inflammation • Long-term immunity can only develop for some 33 . hydrolytic enzymes.

and nystatin • In some cases surgical removal of damaged tissues • Prevention limited to masks and protective clothing to reduce contact with spores 34 . flucytosine. azoles.Control of Mycotic Infections • Immunization is not usually effective • Control involves intravenous amphotericin B.

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and superficial mycoses – Opportunistic mycoses 36 .Organization of Fungal Disease • Mycoses are presented according to type. and degree of pathogenicity – True pathogens: systemic. level of infection. cutaneous.

Levels of invasion by fungal pathogens 37 .

Systemic Infections by True Pathogens • Restricted to endemic regions of the world • Infection occurs when matter containing conidia is disturbed • Spores germinate in the lungs • Infection can become systemic • Spores may be inoculated into the skin • All diseases result in immunity 38 .

ketoconazole 39 . most prevalent in eastern and central regions of U. • Grows in moist soil high in nitrogen content • Inhaled conidia produce primary pulmonary infection that may progress to systemic involvement of a variety of organs and chronic lung disease • Amphotericin B.S. causes histoplasmosis • Typically dimorphic • Distributed worldwide.Histoplasmosis: Ohio Valley Fever • Histoplasma capsulatum – most common true pathogen.

Dimorphic colonies of Histoplasma capsulatum 40 .

Events in Histoplasma infection and histoplasmosis 41 .

and can form nodules in the lungs • Amphotericin B treatment 42 .Coccidioidomycosis: Valley Fever • Coccidioides immitis – causative agent • Distinctive morphology – blocklike arthroconidia in the free-living stage and spherules containing endospores in the lungs • Lives in alkaline soils in semiarid. creates spherules. hot climates and is endemic to southwestern U.S. • Arthrospores inhaled from dust.

Events in Coccidioides infection 43 .

Disseminated coccidioidomycosis 44 .

bone. • Inhaled 10-100 conidia convert to yeasts and multiply in lungs • Symptoms include cough and fever • Chronic cutaneous.S. and nervous system complications • Amphotericin B 45 .Blastomyces Dermatitidis: North American Blastomycosis • Blastomyces dermatitidis – causes blastomycosis • Dimorphic • Free-living species distributed in soil of a large section of the midwestern and southeastern U.

The dimorphic nature of Blastomyces dermatitidis 46 .

Cutaneous blastomycosis in the hand and wrist 47 .

Paracoccidioidomycosis • Paracoccidioides brasiliensis • Distributed in Central and South America • Lung infection occurs through inhalation or inoculation of spores • Systemic disease is not common • Ketoconazole. sulfa drugs The morphology of Paracoccidioides 48 . amphotericin B.

they can invade • Most species in this group are greatly inhibited by higher temperatures of the blood and viscera • Diseases are progressive 49 .Subcutaneous Mycoses • Subcutaneous mycoses: when fungi are transferred directly into traumatized skin.

then spreads to nearby lymph nodes 50 .Sporothrix Schenckii • Sporotrichosis (rose-gardener’s disease) • Very common saprobic fungus that decomposes plant matter in soil • Infects appendages and lungs • Lymphocutaneous variety occurs when contaminated plant matter penetrates the skin and the pathogen forms a nodule.

The microscopic morphology of Sporothrix schenckii Clinical appearance of lymphocutaneous sporotrichosis 51 .

Phialophora verrucosa. yeast-like bodies. thick. sclerotic cells • Phaeohyphomycosis differs in the causative species and the appearance of the infectious agent 52 .Chromoblastomycosis and Phaeohyphomycosis • Chromoblastomycosis: A progressive subcutaneous mycosis characterized by highly visible verrucous lesions – Etiologic agents are soil saprobes with dark-pigmented mycelia and spores – Fonsecaea pedrosoi. Cladosporium carrionii – Produce very large.

tumor-like disease of the hand or foot due to chronic fungal infection.Mycetoma • When soil microbes are accidentally implanted into the skin • Progressive. may lead to loss of body part • Caused by Pseudallescheria or Madurella Mycetoma caused by Madurella 53 .

Epidermophyton • Closely related and morphologically similar • Causative agent of ring worm varies case to case 54 . hair. Microsporum. nails) are called dermatophytoses – ringworm and tinea • 39 species in the genera Trichophyton.Cutaneous Mycoses • Infections strictly confined to keratinized epidermis (skin.

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and intimate contact promote infection • Long infection period followed by localized inflammation and allergic reactions to fungal proteins 56 . animals. presence of abraded skin.• Natural reservoirs – humans. and soil • Hardiness of the dermatophyte spores.

red ring lesions anywhere on smooth skin • Ringworm of groin (tinea cruris) “jock itch” affects groin and scrotal regions 57 . hair may be lost • Ringworm of beard (tinea barbae) affects the chin and beard of adult males.• Ringworm of scalp (tinea capitis) affects scalp and hair-bearing regions of head. contracted mainly from animals • Ringworm of body (tinea corporis) occurs as inflamed.

• Ringworm of foot and hand (tinea pedis and tinea manuum) is spread by exposure to public surfaces. occurs between digits and on soles • Ringworm of nails (tinea unguium) is a persistent colonization of the nails of the hands and feet that distorts the nail bed • Treatment of dermatophytes includes topical antifungal agents – tolnaftate. miconazole applied for several weeks • Lamisil or griseofulvin 1-2 years 58 .

Ringworm lesions Ringworm of the extremities 59 .

or axillary hair • Black piedra – caused by Piedraia hortae. pubic. mainly on scalp hairs 60 . whitish or colored masses develop scalp. and seborrheic dermatitis • White piedra – caused by Trichosporon beigelii. dark-brown to black gritty nodules. chronic scaling. also implicated in folliculitis. psoriasis. mottling of skin. elicits mild.Superficial Mycoses • Tinea versicolor – caused by Malassezia furfur.

Opportunistic Mycoses • All have predisposing factors Candida – dominant opportunistic pathogen Aspergillus – accounts for most lung infections Cryptococcus Alternaria Paecilomyces Fusarium Rhizopus Torulopsis 61 .

genitalia. fatal systemic diseases • Budding cells of varying size that may form both elongate pseudohyphae and true hyphae • Forms off-white.• Candida albicans • Widespread yeast • Infections can be short-lived. superficial skin irritations to overwhelming. pasty colony with a yeasty odor Infections by Candida: Candidiasis Candida Albicans • Normal flora of oral cavity. adherent growth on the mucous membranes of mouth and throat • Vulvovaginal yeast infection – painful inflammatory condition of the female genital region that causes ulceration and discharge • Cutaneous candidiasis – occurs in chronically moist areas of skin and in burn patients 62 . large intestine or skin of 20% of humans • Account for 70% of nosocomial fungal infections • Thrush – occurs as a thick. white.

Infections by Candida albicans Detection of Candida albicans 63 .

Diagnosis and Treatment • Presumptive diagnosis made if budding yeast cells and pseudohyphae are found. amphotericin B and fluconazole for systemics 64 . germ tube • Growth on selective. differential media differentiates Candida species • Topical antifungals for superficial infections.

or diabetes patients • Infection of lungs leads to cough. cancer. and lung nodules • Dissemination to meninges and brain can cause severe neurological disturbance and death 65 . fever.Cryptococcosis and Cryptococcus Neoformans • Cryptococcus neoformans causes cryptococcosis • A widespread encapsulated yeast that inhabits soil around pigeon roosts • Common infection of AIDS.

Diagnosis and Treatment • Negative stain demonstrating encapsulated budding yeast • Biochemical tests. serological testing • Systemic infection requires amphotericin B and fluconazole 66 .

unicellular fungus that causes pneumonia (PCP). the most prominent opportunistic infection in AIDS patients • This pneumonia forms secretions in the lungs that block breathing and can be rapidly fatal if not controlled with medication • Pentamidine and cotrimoxazole 67 .Pneumocystis (Carinii) Jiroveci and Pneumocystis Pneumonia • A small.

and transplant patients • Infection usually occurs in lungs – spores germinate in lungs and form fungal balls. and conjunctiva • Invasive aspergillosis can produce necrotic pneumonia. heart. fumigatus most commonly • Serious opportunistic threat to AIDS. and other organs • Amphotericin B and nystatin 68 . and infection of brain. eyelids. A. leukemia. ear canals. can colonize sinuses. 8 involved in human disease.Aspergillosis: Diseases of the Genus Aspergillus • Very common airborne soil fungus • 600 species.

Clinical aspects of Aspergillosis Microscopic appearance of Aspergillus 69 .

and brain of people with diabetes and malnutrition. organic debris. and Mucor • Usually harmless air contaminants invade the membranes of the nose. heart.Zygomycosis • Zygomycota are extremely abundant saprobic fungi found in soil. eyes. water. Absidia. and food • Genera most often involved are Rhizopus. with severe consequences Absidia corymbifera 70 .

toenails. occasionally infects eyes. burned skin 71 .Miscellaneous Opportunists • Any fungus can be implicated in infections when immune defenses are severely compromised • Geotrichum candidum – geotrichosis. dairy products. mold found in soil. primarily involved in secondary lung infections • Fusarium species – soil.

grains. peanuts. severe hematologic and neurological damage 72 Fungal Allergies and Intoxications . corn. lethal to poultry and livestock Stachybotrys chartarum – sick building syndrome. teapicker’s lung. bark stripper’s disease Fungal toxins lead to mycotoxicoses usually caused by ingesting or inhaling fungal toxins – Aflatoxin toxic and carcinogenic.Common fungi that can cause uncommon infections • • • • Fungal spores are common sources of atopic allergies Seasonal allergies and asthma – Farmer’s lung.