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Mycology

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Mycology Introduction Student Lab Division of Medical Technology Carol Larson MSEd, MT(ASCP) Mycoses Superficial Subcutaneous Systemic Opportunistic ... – PowerPoint PPT presentation

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Title: Mycology


1
Mycology Introduction
  • Student Lab
  • Division of Medical Technology
  • Carol Larson MSEd, MT(ASCP)

2
Mycoses
  • Superficial
  • Subcutaneous
  • Systemic
  • Opportunistic

3
Characteristics of fungi
  • Eukaryotic
  • Growth requirements
  • Forms
  • Mold
  • Yeast

4
Hyphae
  • Septate
  • Aseptate

5
Hyphae
  • Hyaline
  • Dematiaceous

6
Mycelium
  • Mass of branching intertwined hyphae
  • Vegetative
  • Aerial
  • Fertile

7
Vegetative types
  • Favic chandeliers
  • Nodular organs
  • Racquet hyphae
  • Spiral hyphae

8
Reproduction
  • Identify fungi by
  • Morphology of reproductive structures
  • Spores from vegetative mycelium or aerial
    fruiting bodies

9
Asexual Reproduction
  • Conidia
  • Conidiophore
  • Arthroconidia

10
Asexual Reproduction
  • Blastoconidia
  • Pseudohyphae
  • Chlamydoconidia
  • Chlamydospores

11
Asexual Reproduction
  • Macroconidia
  • Microconidia
  • Phialoconidia
  • Phialide

12
Asexual Reproduction
  • Annelloconidia
  • Annellide
  • Sporangiospores
  • Sporangium
  • Sporangiophore

13
Sexual Reproduction
  • Perfect Fungi has a sexual stage
  • Fungi Imperfecti no know sexual stage
  • Spores

14
Sexual Reproduction
  • Ascospores
  • Ascus
  • Ascocarp
  • Basidiospores
  • Zygospores

15
In review
  • Mycoses fungal diseases
  • Characteristics of fungi
  • Growth requirements
  • Forms (mold, yeast)
  • Structures
  • Reproduction
  • Asexual
  • Sexual

16
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

17
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

18
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

19
Direct Examination
  • Saline wet mount
  • Lactophenol cotton blue wet mount
  • 10 KOH preparation
  • Gram stain
  • Acid fast stain
  • India ink stain

20
Direct Examination
  • Calcofluor white stain
  • Wrights stain
  • Gomori Methenamine Silver stain
  • Periodic Acid Schiff stain

21
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

22
Specimen Processing
  • Primary isolation media
  • Goal isolate potential pathogens
  • Use non-selective and selective media
  • Proper ingredients
  • Incubation temperature
  • Incubation time
  • Incubation atmosphere

23
Non-selective Media
  • Sabouraud dextrose agar
  • Brain heart infusion (BHI) with/without 5 blood
    and 1 glucose

24
Selective Media
  • Mycosel agar
  • Inhibitory mold agar
  • Dermatophyte test medium

25
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

26
Examination of Culture Growth
  • Potential pathogens
  • Slow growers
  • Growth on Mycosel
  • Color dull buff, brown, mousy gray
  • Dimorphic

27
Examination of Culture Growth
  • Growth rate
  • Rapid growers 1-5 days
  • Intermediate growers 6-10 days
  • Slow growers gt10 days

28
Colony Morphology Appearance
  • Rugose
  • Umbonate
  • Verrucose
  • Flat

29
Colony Morphology Texture
  • Cottony
  • Glabrous
  • Granular
  • Velvety

30
Colony Morphology Pigmentation
  • Surface
  • Reverse

31
Microscopic Morphology
  • Definitive means of identification
  • Evaluate
  • Shape
  • Method of production
  • Arrangement of conidia/spores
  • Size and color of hyphae

32
Microscopic Techniques
  • Tease mount
  • Scotch tape preparation
  • Slide culture

33
Serological Diagnosis
  • Immunodiffusion
  • Complement fixation
  • EIA
  • Latex agglutination

34
Antifungal Susceptibility
  • Determine appropriateness
  • Standardization of testing
  • Methods
  • Predictability in vivo
  • Antifungal agents

35
In Summary
  • Specimen collection and transport
  • Specimen processing and culture
  • Direct examination of specimen
  • Examination of culture
  • Serological testing
  • Antifungal susceptibility
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