2024 In-House Review Edition - Medical Mycology PDF

2024 IN-HOUSE REVIEW EDITION REVIEW MODULE IN MEDICAL MYCOLOGY PROF. JOSHUA ANGELO HERMIDA MANDANAS International Tropical Medicine Scientist National Medical Technolo...

2024 IN-HOUSE REVIEW EDITION REVIEW MODULE IN MEDICAL MYCOLOGY PROF. JOSHUA ANGELO HERMIDA MANDANAS International Tropical Medicine Scientist National Medical Technology Board Reviewer General characteristics of fungi:  Aerobic  Nucleated  Achlorophyllous  Cell wall (chitin-N-acetyl-D-glucosamine (B1->4) units)  Eukaryotic  Neutral pH 1 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Introduction to Mycological Terminologies Yeast  unicellular organisms with round/oval in shape and size ranges from (2-60 um)  reproduce asexually by budding/blastospores  Reproduce sexually by meiosis Hyphae  Long,slender branching tubes  Basic structural unit of molds  Mass of hyphae is called mycelium  Septate: has crosswalls  Aseptate/Coenocytic: absence of cross walls Spiral: tightly coiled Rhizoids: rootlike structures Vegetative hyphae: body of fungi Vegetative mycelium: nutrient absorbing & water exchanging part of fungus Aerial hyphae: portion extending above substrate surface Spores  Functionally similar to seeds of higher plants  Sexual (karyogamy & Meiosis)  Asexual (mitosis): most common, Sporangium: spores produced in sac like structure conidia: spores produced at tips/sides of hyphae Reproduction of Fungi Asexual ( imperfect): formation of conidia Types of conidia 1. Phialides: vaselike structures 2. Annellides: ringed structures 3. Arthroconidia: fragmentation of fertile hyphae (Trichosporon beigelii, Aureobasidium, Coccidioides immitis,Geotrichum candidum) Sexual (telomorph): joining of 2 compatible nuclei, followed by meiosis Asexual (anamorph) ex. Pseudoallescheria boydii anamorphs: Scedosporium boydii & Graphium Fungal Pigmentation Hyaline/moniliaceous: nonpigmented Phaeoid/dematiaceous: darkly pigmented Stains 1. Gomori Methylene Stain: all fungi stains black 2. Mason-Fontana: stains melanin hyaline (pink/red) phaeoid (brown) Forms of fungi Dimorphic: includes mold phase at RT & yeast/spherule phase (37°C + Co2) ex. Blastomyces dermatitidis, Coccidioides immitis, Histoplasma capsulatum, Paracoccidioides braziliensis, Sporothrix schenckii, Penicillium marneffei Polymorphic fungi: both yeast & mold forms in ame culture ex. Exophiala SYSTEMIC DERMATOPHYTE/ CUTANEOUS MYCOSES General characteristic  Keratinophillic  Deep infections are rare  Reproductive cells: Microconidia & Macroconidia  Athlete’s foot: “moccasin foot” 2 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Ringworm infection Site affected Tinea capitis Head Tinea favosa Head Tinea barbae Beard Tinea corporis Body Tinea manuum Hand Tinea unguium Nails Tinea cruris Groin Tinea pedis Feet Tinea imbricata Body o Microsporum canis Infection: hair & skin, erythema Macroconidia: spindle shaped, echinulate, thick walled, tapering spiny distal ends Colony: fluffy & white reverse (lemon yellow)- PDA Note: Positive fluorescence in wood’s lamp o Microsporum gypseum Infection: tinea barbae Macroconidia: fusiform, thin walled, spindle shaped (4-6 septa) Colony: powdery cinnamon color, develops white tufts of sterile hyphae in aging cultures (pleomorphic) beneath: brown/red pigment o Microsporum audounii Infection: gray patch tinea capitis (decades ago) Colony: chlamydoconidium-like swellings, cottony white, no growth in rice media reverse: little/no pigment o Microsporum ferrugineum Infection: gray patch tinea capitis Colony: “bamboo hyphae” (1) suede like-yellow (2) leathery white o Epidermophyton floccosum Macroconidia: smooth, thin walled Infection: skin & nails Colony: yellow to yellowish green (olive drab), suede leather appearance Note: “beaver’s tail-like-conidia” , Dutch pants fuseaux o Trichophyton concentricum Infection: tinea imbricata (form of tinea corporis) Colony: velvety in appearance, wrinkled Note: thiamine enhances recovery o Trichophyton mentagrophytes Microconidia: grape like (“en grappe”) Macroconidia: thin walled, smooth & cigar shaped Infection: athlete’s foot, tinea (capitis, corporis, barbae, cruris) Colony: (1) “downy” (2) granular reverse: yellow Note: Positive for Hair baiting/penetration test Positive for urease o Trichophyton rubrum Infection: foot & toenails Macroconidia: 3-8 cells, cylindric, pencil shaped, Microconidia: clavate/peg shaped, teardrop shaped Colony: surface- white reverse- red 3 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION o Trichophyton tonsurans Infection: “black dot” tinea capitis , “adult ringworm of scalp” Microconidia: “balloon-shaped” Colony: yellowish brown (surface) rust color (reverse) Note: thiamine enhances growth o Trichophyton verrucosum Infection: tinea corporis, usually found in cattles o Trichophyton schoenleinii Infection: Tinea Favosa/Favus (chronic hair inf.-cup shaped flakes called scutula) Colony: Favic chandelier hyphae Note: requires thiamine and inositol SUPERFICIAL MYCOSES General Characteristics  Affects only stratum corneum  Do not have overt symptoms  Fungal agents do not elicit immune response o Malassezia furfur Infection: tinea versicolor “fawn-colored liver spots”, Ptyriasis versicolor ,dandruff, patients with lipid replacement therapy, high corticosteroid therapy Colony: “spaghetti & meatballs fungus” (septate with branched hyphal elements and budding yeasts)-KOH Note: requires lipids (triglycerides, saturated fatty acids) for growth yellow fluorescence in Wood’s Lamp o Hortaea werneckii (Phaeoannellomyces/Exophiala) Infection: tinea nigra (confused with melanoma) Colony: color (shiny,moist colonies with brownish coloration) young – budding blastoconidia old- clustered blastoconidia, annelloconidia Piedra agents A.k.a “ Tinea nodosa”, “Chignon disease’, “Beigel’s disease” o Piedra hortae Infection: black piedra (ascospore)-KOH Colony: brown, thick walled rhomboid cells o Trichosporon beigelii Infection: white piedra (septate hyphae->arthrospore)-KOH Colony: produces arthroconidia, hyphae & blastoconidia, strawberry color, velvety in appearance Note: negative carbohydrate fermentation positive urease assimilation of sugars potassium nitrate o Keratitis fungal agents Fungi involved: Fusarium, Aspergillus, Curvularia, Penicillium, Cephalosporium,Candida spp Fusarium: Sickle/canoe shaped, multiseptate macroconidia o Otomycosis/Fungus ear/Otitis externa Fungi involved: Aspergillus, Mucor, Penicillium, Rhizopus SUBCUTANEOUS MYCOSES  Mode of transmission: skin trauma/prick  Normal habitat: soil/ decaying vegetation  Chronic infection o Sporothrix schenkii Infection: Gardener’s disease, Rose Handler’s disease 4 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Colony morphology: (1)mold form- daisy-like,rosette, flowerette (brown to black greasy appearance) (2) yeast form (BHI + SRBC + 37°C + CO2)- cigar bodies, asteroid Note: grows in medium with cycloheximide o Phaeohyphomycosis Caused by darkly pigmented fungi Colony: tissue- forms yeastlike cells & irregular/beaded septate hyphae Organisms: Dreschlera spicifera (most common), Alternaria, Bipolaris, Chaetomium, Cladosporium, Curvularia, Dactylaria,Exophiala, Fonsecaea, Mycocentrospora, Ochronosis, Oidiodendron, Phaeosclera, Phialophora, Phoma, Ulocladium, Wangiella,Xylohypha o Chromoblastomycosis A.k.a Chromomycosis, Verrucous dermatitis Infection: chronic skin infection (cauliflower like surface), sclerotic bodies Phaeoid fungi o Phialophora verrucosa Colony: Vase-like/phialides, flask shaped with collarettes o Fonsecaea pedrosoi Colony:Short chain/well defined septa, longer conidiophores & spores o Fonsecaea compactum similar to F. pedrosoi but with more compact conidial heads o Cladophialophora carrionii Colonies: Dark with jet black reverse,Long branching chains, erect conidiophores Infected tissue: Medlar bodies Shield cell- conidia at tip of conidiophore o Mycetoma Agents  A.k.a Madura foot, maduromycosis  Swellings with characteristic exudate draining skin surface  Also known as Maduromycosis, Madura foot (1800s)  Chronic, progressive, infectious disease of skin/subcutaneous tissue characterized by formation of tumor like swellings and grains  Usually caused by bacteria (Actinomycetes, Streptomyces) or fungi (Madurella) nonseptate hypahe: tissue form Actinomycotic: Actinomadurae, Nocardia, Streptomyces Eumycotic agents: 1.) Pseudoallescheria boydii Amorphic form: Scedosporium boydii Colony: annelids (oval conidia at conidiogenous cells) Teleomorph: formation of cleistothecia Note: homothallic 2.) Acremonium falciforme Colony: mucoid clusters of conidia 3.) Madurella Colony: septate, sterile conidia, characteristic diffusible brown pigment o Rhinosporidium seeberi Infection: hyperplasia of mucous membranes/polypoid masses (swimming) Colony: large spherical sporangia (tissue form) o Lacazia loboi A.k.a Loboa loboi Infection: lobomycosis (keloid like nodules) Tissue form: Multiple budding cells 5 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION SYSTEMIC MYCOSES  Dimorphic  Primary infection is pulmonary, it can disseminate into organs  mold phase of B.dermatitidis, C. immitis, H. capsulatum, P.braziliensis requires confirmatory by DNA probe o Blastomyces dermatitidis Infection: (middle-aged men) Gilchrist’s disease, Chicago disease, North American blastomycosis, affects lungs,liver & skin Telomorph/Sexual stage: Ajellomyces dermatitidis Colony: RT- pyriform spores,dumbbell shaped, raised spicules, lollipop appearance 37°C(KOH)- broad based budding yeast cells, large yeast (8-12 um) Culture media: Blood agar, 37°C o Paracoccidioides brasiliensis Infection: south american blastomycosis, “para”, “paracocci”,Brazilian blastomycosis, Paracoccidioidal granuloma, Lutz Splendore-Almeida disease, affects skin, mucous membranes, lymph nodes & internal organs, “Arise of Tutankhamen’s tomb” Colony: mold form- lollipop appearance yeast form- Mariner’s wheel, thick walled multiple buds Resembles B.dermatitidis (daughter yeast cells) o Histoplasma capsulatum Infection: infective stage (microconidia), histoplasmosis, reticuloendothelial cytomycosis, cave disease, spelunker’s disease, darling’s disease Medium for transmission: feces (starling’s, chicken & bats) Note: resides in soil with high nitrogen content Telemorph/Sexual stage: Ajellomyces capsulatus, Emmonsiella capsulata Mold form: tuberculate microconidia Yeast form: intracellular yeast Histoplasma capsulatum var. duboisii: african histoplasmosis ( skin, lymph nodes & bone) Resembles Chrysosporium spp/ (microconidia) Sepedonium spp.(macroconidia) Culture media: Pines medium, glucose-cysteine blood, BHI-blood, 37°C o Coccidioides immitis Infection: infective stage (arthroconidia), desert rheumatism, erythema multiforme, erythema nodosum, toxic erythema, “Cocco”, desert fever, valley fever, “the bumps”,”San Joaquin Valley fever” Mold form: barrel shaped arthroconidia Yeast form: spherule with endospores ( spherule resembles B.dermatitidis) ( endospores resemble C. neoformans, H. capsulatum, P. brasiliensis) o Penicillium marneffei Infection: systemic-fatal (immunocompromised) Mold form: green aerial & vegetative hyphae, red diffusable pigment Yeast form: cross walled (resembles H. capsulatum), sausage like appearance Exoantigen Test A antigen = Blastomyces dermatitidis HS, HL , F = Coccidioides immitis H & M antigens = Histoplasma capsulatum 1,2,3 antigens= Paracoccidioides brasiliensis OPPORTUNISTIC MYCOSES  Normal flora  Immunocompromised o Zygomycota/ Mucorales/Mucormycosis Mode of transmission: inhalation of SPORANGIOSPORES Habitat: soil Infection: opportunistic in immunocompromised 6 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Sexual reproduction: zygospores Asexual reproduction: sporangiospores 1. Lichtheimia (formerly Absidia) infection: rhinocerebral zygomycosis, diabetic patients with ketoacidosis, thrombosis & necrosis 2. Mucor Infection: rhinocerebral zygomycosis Colony: no rhizoids 3. Rhizopus: most common Infection: rhinocerebral mucormycosis in diabetic patients, refractory to treatment Colony: rapid growers, wolly colonies, umbrella shaped structure, brown rhizoids connected by STOLONS 4. Syncephalastrum Colony: confused with aspergillus Has large columella with merosporangiam found in decaying vegetation 5. Cunninghamella Infection: recovered from sinus (disseminated) Colony: erect sporangiophores that branch into sporangioles with fine spines o Aspergillus Infection: neutropenia (single most predisposing factor) conidia (infective stage) bone marrow transplant, affects lungs (pneumonia like symptoms),fungus ball, steroids injection (Exserohilum rostratum) Colony: hyaline septate hyphae, “bread mold”, dichotomously branched hyphae uniserate- phialides attach directly to conidiophore biserate- supporting structure called metula Aspergillus fumigatus: light green color to blue o Cryptococcus (Filobasidiella) neoformans Infection: menigitis, pulmonary disease & septicemia (AIDS), torulosis, European blastomycosis Sexual stage: FILOBASIDIELLA Colony: mucoid, produces blastoconidia only Detection: old (capsule)- india ink new (Ag)- latex agglutination Key reactions: grows at 37°C, sensitive to cycloheximide, positive urease,negative nitrate, production of phenol oxidase, positive for phospholipase, grows in birdseed agar Canavanine glycine bromthymol blue agar: differentiates Crytptococcus gattii (positive) from Cryptococcus neoformans o Rhodotorula Colony: bright salmon pink Resembles Cryptococcus neoformans o Candida spp. Infection: wide (superficial – disseminated), thrush, nail infection, mycotic vulvovaginitis Culture: PAS stain, GMS, Urease negative (except C.krusei), Assimilation tests, Chlamydospore/Cornmeal agar(yeast cells & pseudohyphae- candida spp.), germ tube (only C. albicans, C.dubliniensis, C. stellatoidea) o Pneumocystis spp. P. carinii: rats P. jiroveci: humans Infection: interstitial plasma cell pneumonia (premature infants/AIDS) Infective stage: cysts Colony: nonfilamentous fungus Life stages: Trophozoite -> Precyst -> Cyst Diagnosis GMS (punched out ping pong ball appearance) Masson-Fontana silver stain Mucicarmine 7 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Term Definition Aerial mycelium Hyphae produced above agar media surface Anamorph Asexual form of fungal sporulation; imperfect state Arthroconidium Conidium derived from fragmentation of specialized hyphae Ascocarp/Ascoma Fruiting structure that contains asci Ascospore Sexual spore formed within an ascus following meiosis Ascus Sac like structure that contains ascospores, characteristic of Ascomycetes Basidiospore Sexual spore formed as an outgrowth of basidium Basidium Structure that contains basidiospores Blastoconidium Asexual spore formed by budding of yeast cell Chlamydospore/Chlamydoconidia Thick walled resting/ survival structure Cleistothecum Enclosed ascocarp, composed of layers of hyphae that contains randomly dispersed asci Collarette Funnel shaped structure at apex of phialide Columella An extension of sporangiophore into base of sporangium Conidiogenous cell Cell that produces conidia Conidiophore Specialized hyphal structure that carries conidia Conidium Asexual reproductive structure formed in any manner that does not involve cleavage Dematiaceous Dark colored pigmented Dimorphic Displaying two morphologic types, one environmental (mold) and one in vivo (yeast) Glabrous Smooth colonial morphology Heterothallic Sexual reproduction requiring interaction of two different thalli Holomorph Whole fungus; anamorphic plus telemorphic state of fungus Homothallic Sexual reproduction can take place within one thallus Hyaline Colorless/transparent Hyphae Vegetative unit of mold Intercalary Borne within hypha Macroconidium Larger of two types of conidia produced by mold Microconidium Smaller of 2 types of conidia produced by mold Mold Filamentous fungi that reproduces by sexual/asexual means Mycelium Mass of hyphae that makes up the thallus Perithecium Enclosed ascocarp with pore at top through which ascospores are discharged Phialide Cells with opening through which conidia are produced Pseudohyphae Connected yeast cells (blastoconidia) that resembles hyphae but contains areas of constriction between adjacent cells Scleotium Multi cellular clump of cells that does not produce any spores or conidia Septum Cross walls of hyphae Sporangiophore Stalk bearing sporangium Sporangiospore Asexual spore produced within sporangium Sporangium Sac like structure in which asexual sporangiospores develop Sterigmata Slender outgrowth of cell bearing conidia Teleomorph Sexual form of fungal sporulation Terminal Borne at end of hyphae Thallus Vegetative growth of fungus, includes an interwoven mass of hyphae 8 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Thermophillic Fungi that grows at high temperatures Vegetative mycelium Hyphae produced on surface or extending in agar media Vesicle Enlarged/swollen cell, often at end of conidiiophore or sporangiophore, which may also be within hyphae Yeast Single celled fungus that reproduces by budding or by fission Rapid Testing for Presumptive Fungi isolate Identification Urease production Cryptococcus neoformans Germ tube production Candida albicans/dubliniensis Pseudohyphae present Candida spp. Chlamydoconidia present Candida albicans/dubliensis Lipid growth requirement Malassezia furfur species complex Red colonial pigmentation Rhodotorula species Ascopore formation Saccharomyces cerevisiae Trehalose assimilation Candida glabrata Medium Purpose Ascospore medium, V8 agar Ascopspore development Birdseed (niger) agar Demonstration of melanin in Cryptococcus neoformans Brain Heart Infusion with blood Enhances recovery of dimorphic fungi Christensen Urea Agar Differentiation of Trichophyton rubrum and Trichophyton mentagrophytes CHROMagar Candida Chromogenic mixture to identify yeast species Cornmeal agar with Tween 80 Visualization of yeast morphology Czapek Dox agar Identification of Aspergillus species Potato Dextrose Agar Pigment development of Trichophyton rubrum and mold morphology Sabouraud Dextrose agar General purpose fungal growth medium (traditional formulation or Emmon’s modification) SAB agar with cycloheximide Selective medium to inhibit saprophytic fungi (antibacterials such as chloramphenicol or gentamicin are included) Slide culture (various agars) Study of microscopic structure Trichophyton agar Differentiation of Trichophyton species Gene target Fungal Specie Actin Candida albicans B-tubulin Aspergillus species Calmodulin Aspergillus species, Fusarium species Chitin synthetase Dermatophyte species Cytochrome b Candida albicans Dihydrofolate reductase Pneumocystis jiroveci EF-1 Fusarium species Mitochondrial rRNA Pneumocystis jiroveci rDNA complex ( 5S gene) Candida albicans, Pneumocystis jiroveci rDNA complex (28S gene) Aspergillus specie, Candida specie rDNA comlex (IGS region) Aspergillus fumigatus SEVERAL ADVANCEMENTS IN MEDICAL MYCOLOGY  mushrooms (hypoglycemic, anti cancer, antipathogenic, enhancer of immune system, vitamin d production)  B-glucan: most abundant fungal cell wall polysaccharide, has immunity-stimulating effects  isolation of penicillin (Penicillium spp. mold), lovastatin (Aspergillus terreus), cyclosporine (Trichoderma polysporum), cephalosporin (Cephalosporium spp.), anti cancer (Aspergillus fumigatus 9 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Compilation of schematic illustrations and tables in Medical Mycology 10 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Candida albicans Budding yeasts and pseudohypahe of C. albicans Aspergillus fumigatus Septate hyphae Nonseptate/Aseptate hyphae 11 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Sporangiospores of Rhizopus spp. Coccidioides’ arthroconidia Hyphae, micro- and macroconidia of T.mentagrophytes Ascospore of Saccharomyces cerevisiae 12 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Germ tubes of Candida albicans Cleistothecium of Pseudoallescheria boydii Tuberculate macroconidia with microconidia of Histoplasma capsulatum Budding yeast cells of Blastomyces dermatitidis 13 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Conidia of Sporothrix schenckii Colonies of Rhizopus spp. Colony and “fruiting head” of Aspergillus flavus Cysts of Pneumocystis jiroveci 14 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION Colony and macroconidia of Microsporum canis Colony of Trichophyton rubrum Colony of Epidermophyton flocossum LABORATORY NOTES IN MEDICAL MYCOLOGY  BSL-3: Coccidioides spp. and H. capsulatum.  Culture plates if used can be flooded with 4% formaldehyde solution (10% formalin) and left at room temperature for several hours or overnight to sterilize the culture before microscopic examination is performed  Potentially airborne: Coccidioides spp. (lung cavity)  The simplest method for direct examination of a specimen is to observe a suspension placed on a slide and coverslip under reduced light. For those specimens that are dry or viscous, the addition of a wetting agent such as saline is necessary prior to observation.  For specimens that contain distracting tissue debris and cells, such as vaginal secretions, nails, and skin scrapings, a solution of potassium hydroxide (KOH) may be used as ameans to dissolve the tissue material, so that fungal elements are more visible 15 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo  2024 IN-HOUSE REVIEW EDITION  The polysaccharide capsule of C. neoformans can be demonstrated by negative staining using India ink particles; it is especially evident when infected cerebrospinal fluid is examined. When examined with the light microscope, the capsule stands out as a clear space around the fungal cell, with ink particles bouncing off the edge as brownian motion occurs, see image below…  Calcofluor white stain: whitener used in the textile and paper industries, this binds to the chitin in the walls of fungal cells and fluoresces white or apple green (depending on the filter combination used) when exposed to short-wavelength ultraviolet light from a fluorescence microscope  Most moulds grow best at 25°–30° C. Most yeasts, however, grow well at 35°–37° C and are often recovered first on enriched blood agar plates in the bacteriology laboratory  The most commonly used selective agar employs cycloheximide to inhibit saprophytic fungi and an antibacterial agent (usually chloramphenicol or gentamicin) to inhibit bacteria  For tissue specimens, especially when dimorphic fungi might be the etiologic agent, an enriched agar, such as brain-heart infusion agar with blood and supplemented with antibiotics may be added  The medium used traditionally is Sabouraud dextrose agar, which has a pH of 5.5–5.6 and was designed for the isolation of dermatophytic fungi  Emmons’ modification of Sabouraud dextrose agar, which contains less glucose with a pH of 6.8–7.0, is more widely used in the clinical mycology laboratory as a general growth medium because of its wide application for the culture of fungi  Recovery of fungi from blood requires special attention and can be accomplished by inoculating broth media by using a biphasic broth-agar system or by using the lysis centrifugation technique. Because a high percentage of fungemias are caused by yeast, continuous-read-automated-culture methods have become the mainstay for fungal detection  In general, the pathogenic dimorphic and dematiaceous fungi grow slower, requiring a week or longer for colonies to appear. Rapidly growing mould colonies that appear after prolonged incubation on mycologic media should be questioned as possible contaminants  Germ tube test: Aseptically transfer several colonies of yeast to a 12 × 75-mm test tube containing approximately 0.5 mL of serum (human, fetal calf, bovine, or rabbit)-> Incubate the tube at 35° C for up to 3 hours-> Place one drop of the mixture on a clean glass slide and coverslip-> Examine under high dry (400×) magnification and reduced light for the presence of germ tubes  Tests to detect fungal antigens or metabolic byproducts in serum or other body fluids on the other hand have been shown to be more useful for the diagnosis and management of fungal disease  approaches to species identification of many fungi today should include both morphologic and molecular testing methods #JEREMIAH 29:11 GOD BLESS YOU ALL! 16 Do not upload in any websites/sell/use in tutorial videos/modify Prof. Joshua Angelo