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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
• 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)
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
SYSTEMIC MYCOSES
OPPOTUNISTIC MYCOSES
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