Mycology

Introduction
Student Lab
Division of Medical Technology
Carol Larson MSEd, MT(ASCP)

Mycoses

Superficial
Subcutaneous
Systemic
Opportunistic

Characteristics of fungi
Eukaryotic
Growth requirements
Forms
Mold
Yeast

Hyphae
Septate

Aseptate

Hyphae
Hyaline

Dematiaceous

Mycelium
Mass of branching intertwined hyphae
Vegetative
Aerial
Fertile

Vegetative types
Favic chandeliers
Nodular organs
Racquet hyphae
Spiral hyphae

Reproduction
Identify fungi by:
Morphology of reproductive structures
Spores from vegetative mycelium or
aerial fruiting bodies

Asexual Reproduction
Conidia
Conidiophore

Arthroconidia

Asexual Reproduction
Blastoconidia
Pseudohyphae
Chlamydoconidia
Chlamydospores

Asexual Reproduction
Macroconidia
Microconidia

Phialoconidia
Phialide

Asexual Reproduction
Annelloconidia
Annellide

Sporangiospores
Sporangium
Sporangiophore

Sexual Reproduction
Perfect Fungi has a sexual stage
Fungi Imperfecti no know sexual
stage
Spores

Sexual Reproduction
Ascospores
Ascus
Ascocarp

Basidiospores
Zygospores

In review
Mycoses fungal diseases
Characteristics of fungi
Growth requirements
Forms (mold, yeast)
Structures
Reproduction
Asexual
Sexual

Fungal Culture Process

Specimen collection and transportation

Direct examination of specimen
Selection and inoculation of media
Evaluation of fungal growth
Serological testing
Antifungal susceptibility testing

Specimen Collection

Specimen types
Collect from area most likely infected
Use sterile technique
Keep specimen moist
Label container properly
Transport right away
Process right away

Direct Examination

Provides preliminary report

Guides MD in treatment of patient
Observe yeast phase of dimorphic
Gives clues to id causative agent
Inoculate special media
May require more than one direct
examination method

Direct Examination

Saline wet mount

Lactophenol cotton blue wet mount
10% KOH preparation
Gram stain
Acid fast stain
India ink stain

Direct Examination

Calcofluor white stain

Wrights stain
Gomori Methenamine Silver stain
Periodic Acid Schiff stain

Specimen Processing
Safety
Tube media preferred over plate media
Work in safety hood
Wear gloves and lab coat
Autoclave specimens and media
Disinfect work area daily

Specimen Processing
Primary isolation media
Goal: isolate potential pathogens
Use non-selective and selective media
Proper ingredients
Incubation temperature
Incubation time
Incubation atmosphere

Non-selective Media
Sabouraud dextrose agar
Brain heart infusion (BHI) with/without
5% blood and 1% glucose

Selective Media
Mycosel agar
Inhibitory mold agar
Dermatophyte test medium

Subculture / Identification Media

Neutral Sabouraud dextrose agar
(Emmons)
Cornmeal-Tween 80 agar
Niger seed agar (Birdseed agar)
Tween 80 / Oxgall / caffeic acid agar
Potato dextrose agar

Examination of Culture Growth

Potential pathogens
Slow growers
Growth on Mycosel
Color: dull buff, brown, mousy gray
Dimorphic

Examination of Culture Growth

Growth rate
Rapid growers: 1-5 days
Intermediate growers: 6-10 days
Slow growers: >10 days

Colony Morphology
Appearance

Rugose
Umbonate
Verrucose
Flat

Colony Morphology
Texture

Cottony
Glabrous
Granular
Velvety

Colony Morphology
Pigmentation
Surface
Reverse

Microscopic Morphology
Definitive means of identification
Evaluate:
Shape
Method of production
Arrangement of conidia/spores
Size and color of hyphae

Microscopic Techniques
Tease mount
Scotch tape preparation
Slide culture

Serological Diagnosis

Immunodiffusion
Complement fixation
EIA
Latex agglutination

Antifungal Susceptibility
Determine appropriateness
Standardization of testing
Methods
Predictability in vivo

Antifungal agents

In Summary

Specimen collection and transport

Specimen processing and culture
Direct examination of specimen
Examination of culture
Serological testing
Antifungal susceptibility