Exam 1 Fungus

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Hyalohyphomycosis

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Hyline fungi

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Phaeohyphomycosis

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dematiacious fungi

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Hyphae

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The mycelium

May be vegitative or Aerial (reproductive)

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Conidiogenous cell

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form the conida (spores)

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Telemorph

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Form of sexual reproduction with mitosis and meiosis. Involves the fusion of two gametes.

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Anamorph

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The non-sexual form of fungus, only spreads via mitosis

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Chlamydoconidium

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A food storage vesicle found at the end of the hypha and is noticably enlarged

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Blastoconidium

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Budding of new fungus along the Hypha

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Arthroconidium

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Formation of new colonies by breaking off bits of the Hypha chain

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Fungus Media

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-Brain Heart infusion Agar (BHI)

-Inhibitory Mold Agar (IMA)

-Sabourard Dextrose Agar (SDA) Low pH to -surpress bacteria

-Mycosel, selective.

Molds grow at 30c, and yeasts at 35c, take at least a month to grow

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Antifunguls: Cell walls

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Beta-Glucan Synthase Inhibitors

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Antifungul: Cell Membrane

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Azoles, Triazole, Polyenes, Allylamines

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Antifungul: Intracellular

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Thymidylate synthetase – pyrimidine analogue

Mitotic inhibitor

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Candida Morphology

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Large creamy colonies with blebs

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Candida Pathology

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Normal flora on and within the human body, infections are typically the result overgrowth do to immune failure, prolonged antibiotic uses, ect.

Causes mild rashes, such as diaper rash.

Can become a serious blood born infection for the AIDs patients, also forms thrus in these cases.

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Candida Specimen collection

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Plate onto IMA, Candi-select chromagenic agar, or SBA/CA

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Candida Germ Tube

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Uses Candida albicans complex

-Incubate for 3 hours in rabbit serum at 37c

-Wet mound sample from tube and look for a germination tube, Perpendicular extensions with no constriction

-C. albicans complex is likely to test positive, if so treat with fluconazole

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C. albicans ID

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One bulb at end, growth at 42c

Xyl/Tre/MDG positive

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C. dubliniensis ID

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Three bulbs at the end of pseudohypahe

Dose not grow at 42c

XYL/TRE/MDG negative

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Where is Cryptococcus normally found?

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Aged bird droppings around the world.

Also on Eukolyptus, Oak, Maple and Firs

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Cryptococcus basics

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Non-fementing aerobe

Mucoid creamy tan/pink colonies

Stains gram +, and capsules stain gram-

Single bud with a narrow neck

anti-phagocytic capsule

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Cryptococcus neoformans Pathology

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Those with immune deficencies are at risk of infection.

Causes pneumonia and meningitis if inhaled

Also causes sepsis, skin/bone lesions and can infect many of the body organs

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Cryptococcus Isolation

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Specimine from CSF and serum, sputum is acceptable

Must be processed within 2 hours

Grow on SDA or cornmeal for 2-3 days at 37c

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Cryptococcus testing

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Cryptococcus Antigen Latex Agglutination Test (CALAT)

False positives caused by Rheumantic factor, Trichosporon, Malignancy

Phenoloxidase test

Caffeic acid test, forms brown colonies if positive

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L-canavanine + glycine + bromothymol blue agar

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C. gattii assimilates glycine => color change yellow to blue

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Cryptococcus ID

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C. gatti assimilates glycin, but C. neoformans dose not

Are urease +

Inostiol +

Narrow base budding

Capsules stain with Mucicarmine/india ink

Melanin +

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Cryptococcus Treatment

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Amphotericin B and 5-Flucytosine for two weeks

Oral fluconazole for 8 weeks

Voriconazole

R to Caspofungin

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Malassezia furfur

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Found on the skin

-needs fatty acids to grow, add vergin olive oil to the plate after inoculating

-Causes scaling skin syndrome of various colors

-Glows Red

-Small budding yeast with a distinct colarette between cells

-Direct KOH mount; Hyphal bits in the skin

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Aureobasidium pullulans

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A black yeast

Smooth moist colony

Many single celled round hyline conidia

Causes: Cutaneous phaeohyphomycosis Disseminated nosocomial infections

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Phaeoannellomyces werneckii (= Exophiala werneckii)

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A black yeast

Likely identical with Exophiala jeanselmei

2-celled yeast cells

Annelide appears as deep stained septum separating the cells

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Annelide

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a cell that produces and extrudes conidia; the tip tapers, lengthens, and acquires a ring

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Phialide

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a cell that produces and extrudes conidia without tapering or increasing in length; typically acquires the shape of a flask, vase, or tenpin

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Floccose

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Cottony; raw fuzzy cotton

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Glabrous

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Smooth, Without or almost without aerial hyphae

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Hilum

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Scar of attachment; Appears where the conidium was attached to conidiophore and/or another conidium

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Cleistothecium

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a large, round, and closed multicelled structure in which asci and ascospores are produced – sexual stage

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Huelle cells

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Large sterile cells with a thick wall and small lumen and which are found in the presence of cleistothcia

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Dichotomous

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Branching of hyphae into two equal branches that are each equal in diameter to the hypha from which they originated

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Metula

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Separate structural portion of the conidiophore that supports the phialide (like a pedestal)

Seen in genera Aspergillus, Penicillium & Paecilomyces

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Foot cell

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Base of conidiophore where it merges with the hyphae

Gives impression of foot

Typically seen in Aspergillus spp

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Aspergillus Basics

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Grows rapidly (3-5 days)

Infection occurs by inhaling he conidia

Causes various pulmonary infections: Aspergilloma – fungus ball, these can appear in the lungs and veins, Allergic bronchopulmonary disease , Onycomycosis, Disseminated

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Aspergillus Morphology

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Colony: surface at first white and then any shade of green, yellow, orange, brown, or black – white apron , velvety or cottony, white, goldish, or brown

Vesicle: conidiophore with bulb on top, Globose/subglobose/pyriform/clavate

Conidiation: uniseriate / biseriate (metula), Long chains of conidia / short chains, Conidia emanate from upper part of vesicle / full vesicle

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Aspergillus

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Hyphae: Dichotomous branching (45° angle), Parallel, narrow hyphae, Septate, Hyline – less than 6 µm, Foot cell

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Galactomannan

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Test for invasive aspergillosis

Aspergillus cell wall Sens: 27-81%; spec. 70-90%

false pos.: GM wildely distributed throughout nature – gatrointestinal translocation, Other fungi have it: Penicillium, Cryptococcus, Paracoccidioides, Histoplasma, Bacteria have cross-reactive sugars: E. coli, Staph, Enterococci, Pseudomonas, PipTazo/AmoxClav -> not anymore!

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Aspergillus fumigatus

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Most common cause of dissem. Aspergillosis

Macroscopic morphology • Velvet or powdery • First white, the turns blueishgreenish to grey

Uniseriate; phialides on upper 2/3 of vesicle; long conidia chains are parallel to conidiophore

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Aspergillus niger

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Invasive Aspergillosis Common in ear infections

Macroscopic morphology • Woolly, first at white to yellow, turning black • Reverse white to yellow

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Aspergillus flavus

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Systemic, sinus, ear infections

Produces aflatoxin – carcinogenic – liver cancer (epoxide)

Macroscopic morphology • Velvety, yellow to green or brown • Reverse goldish to red-brown,

Uniseriate and biseriate; conidia cover entire vesicle; point out in all directions

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Aspergillus terreus

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Systemic, nail, skin, eye, ear infections

Macroscopic morphology • Usually velvety, cinnamon-brown • Reverse white to brown

Biseriate; compactly columnar; smaller vesicles; biseriate

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Aspergillus nidulans

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Macroscopic morphology • Velvety, usually green, buff to yellow where cleistothecia form • Reverse purplish red becoming dark

Biseriate; chains are short, columnar; cleistothecia usually present with reddish ascospores, Huelle cells abundant

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Aspergillus clavatus

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see picture

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P. marneffii

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Only pathogenic form of pencillum

¤ Systemic inf (HIV) ¤ SE Asia / Bamboo rat ¤ Red pigment ¤ Dimorphic (BHI at 35°C to convert) ¤ Yeast-like at 37°C – no buds

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Penicillium morphology

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grayish turquoise to dull green, often yellow exudate, Simple or branched conidiophores, Metulae support flask shaped phialides, Phialides are blunt and produce conidia in chains

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Fusarium

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Causes: Contact lenses / eye infections / keratitis, Skin lesions in burn patients, Nail infections, Angioinvasive disease - disseminated, Fungus ball

Grows after 4 days on potato dextros agar

Colony morphology: initially white and cottony or woolly, FusariumWith age lavender or violet – sometimes yellow or orange with light reverse

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Fusarium ID

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Foot cell like Aspergillus

Two types of conidiation: Marcroconidia – canoes or banana boats / sickle shaped, or simple cluster

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Dematiacious (dark) Molds Diseases

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These infections are very rare

Typically infects soft tissue after a traumatic injury where it causes the following conditions: Chromoblastomycosis, Eumycotic mycetoma, Phaeohyphomycosis

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Chromoblastomycosis

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A chronic soft tissue infection that produces large coliflower growths

Dark sclerotic yeast bodies in tissue – brown/reddish, often described as copper pennies or muriform. These induce a granulomatous reaction

This condition is typically caused by Fonsecea pedrosoi

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Eumycotic mycetoma

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A Chronic granulomatous disease with large black tumors and distinct sinus tracts that release a grainy discharge.

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Phaeohyphomycosis

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Generic term for subcutaneous or systemic infections caused by dematiacious fungi

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Dematiacious Specimine collection and plating

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Only accept tissue biopsies, do not take swabs or sputum.

Plate onto: SDA(black colony)/IMA or Nutrition deficient agar

Grow over 2-4 weeks at 20-30c

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Dematiacious Staining

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Hematoxilin-Eosin/GMS/Fontana Masson-melanin

KOH with CaFL/Giemsa/Wrights

LPCB

Look for narrow dark hyphae with little branching.

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Identification by conidiogenous cells and conidiae

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See slide 8

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Fonsecea pedrosi

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Most common cause of chromoblastomycosis world wide (86-96%); also myctoma

Takes two weeks to grow

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Phialophora verrucosa

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Second most common cause of chromoblastomycosis

rarely causes Phaeohyphomycosis

Grows in 7-12 days

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Phialophora richardsiae (vase-like)

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Rare cause of subcutaneous phaeohyphomycosis

Tip of phialides is flat and saucer-shaped

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Phaeoacremonium parasiticum (spine type)

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Subcutaneous phaeohyphomycosis

Arthritis/endocarditis/disseminated infections

Grows in about a week

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Stachybotrys chartarum

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A common lab contaminant

Produces Mycotoxins that have been linked to pulmonary hemorrhage in children

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Wangiella dermatitidis (Exophiala dermatitidis)

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Phaeohyphomycosis – cutaneous and subcutaneous

Predilection for CNS – brain and eye

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Exophiala jeanselmei

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Mycetoma and phaeohyphomycosis

Long, tapered conidiophores that extend from hyphae in a right angle; Sharply pointed tip

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Pseudallescheria boydii Diseases

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Mycetoma ¤ Subcutaneous tissue ¤ Fungus ball ¤ Disseminated: brain / eyes / lungs / sinuses / meninges

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Cladophialophora carrionii

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Chromoblastomycosis

Tissue: Dark sclerotic bodies

Growth: Slow – 18 days

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Cladophialophora bantiana

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Predilection for CNS => cerebral phaeohyphomycosis

Infection possibly through inhalation – BSC!!-no slide cultures

Chromoblastomycosis

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Alternaria

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Mostly a contaminant, but sometimes causes phaeohyphomycosis – subcutaneous tissue/nail/eye/ nasal sinuses

Grows in 5 days

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Ulocladium

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Mostly a contaminant

Grows in 5 days

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Epicoccum

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A common contaminant

Grows in 5 days

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Bipolaris

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Most common cause of allergic sinusitis

Skin/eye/lung/CNS (brain lesions)

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Exerohilum

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Phaeohyphomycosis – sinusitis/subcutaneious tissue/ cornea/skin/disseminated

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Scytalidium spp

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Common cause of nail and skin infections; rarely subcutaneous absess/sinusitis/endophthalmitis/ lymphadenitis/fungemia

Grows in 3 days

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Chaetomium spp.

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Large spiked hyphae extending through the wall of saclike cleistothecia

rapid growth

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Phoma spp.

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Septate hyphae with asexual fruiting bodies = pycnidiae

Release of hyline conidia through ostiole (opening)