Mycology

Mycology
Classification & General
Properties of Fungi
 Objectives
•To Understand about fungi, other
mycological terms
•Distinguish between fungi and
bacteria
•To understand the classification of
fungi
•Know the various methods for lab
diagnosis of infection due to fungi
Heading/Subheading-
•Morphological features
•Classification
•Cultural characters
•Clinical findings
•Lab Diagnosis
•Treatment
 Definitions
• Mycologists--scientists who study fungi
• Mycology--Study of fungi.

• Mycoses--diseases caused by fungi

• Medical Mycology--- is the study of
mycoses of man and their etiologic agents
 Introduction
• Mykes (Greek word) : Mushroom
• Fungi are eukaryotic protista; differ
from bacteria and other prokaryotes.
1. Cell walls containing chitin (rigidity & support),
mannan & other polysaccharides
2. Cytoplasmic membrane contains ergosterols.
3. Possess true nuclei with nuclear membrane &
paired chromosomes.
4. Divide asexually, sexually or by both
5. Unicellular or multicellular
6. Lack chlorophyll
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Fungi differ from bacteria in the following
points:-
Prokaryote Eukaryotes
(Bacteria) (Fungi)

Diameters 1 micron 4-15microns

nuclear membrane No nuclear membrane Nuclear membrane

Chromosomes Single chromosome multiple

Division Binary fission Mitotic division

cytoplasme No organelles Organelles

Cell wall Peptidoglycan Chitin

Cell membrane No ergosterol Ergosterol

Ribosome 70 S 80 S
FUNGAL CELL STRUCTURE
 Fungal cell membrane
• Consist of ergosterol rather than cholesterol
like bacterial cell membrane.
• Ergosterol is the site of action of antifungal
drugs, amphtericin B & azole group
 Introduction
– Chemoheterotrophs
• Require organic compounds for carbon and
energy
– Food industry: bread, alcohol, edible fungi
(mushrooms), cheeses
– Some are deadly
– Adapted to environments hostile to
bacteria
• pH 5
• High osmotic pressure
• Low moisture
• Complex carbohydrates (ex. Lignin from wood)
• Less nitrogen
 Introduction

• Most fungi are obligatory aerobes,

some are facultative anaerobes, but
none are obligatory anaerobes.
 Introduction
• Simplest fungus :- Unicellular budding
yeast

• Hypha :- Elongation of apical cell

produces a tubular, thread like
structure called hypha

• Mycelium :- Tangled mass of hyphae is

called mycelium. Fungi producing
mycelia are called molds or filamentous
fungi.

• Hyphae may be septate or non-septate

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 Mycology
 How do we identify molds in the lab?
 Based on type of hyphae (septate versus non-
septate)
 Based on color of mycelium
 Based on reproductive structures
 Molds may form either sexual or asexual spores
 Sexual spores are formed from the fusion of nuclei from two
opposite mating strains of the same species. They are only
formed under special conditions, but they are used to classify
fungi (more on this later)
 Asexual spores, which are most commonly used in
identification, are formed by the aerial mycelium of a single
organism by mitosis and cell division
Morphological
Systematic
Clinical
Morphological
Classification
 CLASSIFICATION
• Depending on cell morphology
1. Yeasts
2. Yeast like fungi
3. Molds
4. Dimorphic fungi

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1. Yeasts
• Unicellular fungi which
reproduce by budding
• On culture - produce
smooth, creamy
colonies
e. g Cryptococcus
neoformans
(capsulated yeast)

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Yeast colonies
 2. Yeast like fungi
• Grow partly as yeasts and partly as
elongated cells resembling hyphae
which are called pseudo hyphae.
e.g. Candida albicans

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 3.Molds/ Filamentous
fungi
• Form true mycelia &
reproduce by
formation of different
types of spores.
• Vegetative/ aerial
hyphae
e.g. Rhizopus, Mucor

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 4. Dimorphic fungi
• Occur in 2 forms
Molds (Filaments) – 25C (soil)
Yeasts – 37C (in host tissue)
Most fungi causing systemic infections
are dimorphic:
– Histoplasma capsulatum
– Blastomyces dermatidis
– Paracoccidioides brasiliensis
– Coccidioides immitis
– Penicillium marneffei
– Sporothrix schenkii
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Systematic
Classification
 Systematic classification
• Based on sexual spore formation:
4 classes
1. Zygomycetes Perfect fungi
2. Ascomycetes reproduce sexually
3. Basidiomycetes
4. Deuteromycetes (fungi imperfectii)

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 Sexual spores
• Zygospores:
Fungi forming zygospores are called
zygomycetes.
• Ascospores:
Ascospores are carried in ascus.
Fungi forming ascospores are called
ascomycetes.
Zygospores
Ascospores inside an
ascus
 Sexual spores
• Basidiospores:
Basidiospores are carried on basidium.
Fungi forming basidiospores are called
basidiomycetes.
• Deuteromycetes are fungi whose
sexual spores are unknown.
Basidiospores
 FUNGI IMPERFECTI
(DEUTEROMYCETES)

•Most fungi causing human infections belong to this

group.
 Asexual spores
• Blastospores:
Produced by budding of the yeast cells.
• Conidia:
Produced by molds.
May be microconidia or macroconidia.
 Mycology
 Blastospores consist of a bud coming off the
parental cell
 Mycology
 Conidiospores are produced in a chain at the
end of a conidiophore. Unicellular
conidiospores are called microconidia
Microconidia
 Mycology
 Multicelled conidiospores are called
macroconidia
 Asexual spores
• Arthrospores:
Produced by fragmentation of hyphae.
• Chlamydospores:
Rounded thick walled spores produced
by candida fungus.
• Sporangiospores:
Spores formed within a sac called
sporangium. Formed by zygomycetes.
Arthrospores
 Arthrospores are formed by the
fragmentation of septate hyphae
 Chlamydospores are thick walled spores
formed within a hyphae
 Sporangiospores are formed within a sac
(sporangium) at the end of an aerial hyphae
called a sporangiophore
Clinical
Classification
 MYCOTIC DISEASES
(Four Types)
1. Hypersensitivity
– Allergy
2. Mycotoxicosis
– Production of toxin
3. Mycetismus (mushroom poisoning)
– Pre-formed toxin
4. Infection– Mycosis
 Fungal Diseases
 Mycosis - fungal infection
 Mode of entry & infective tissue:
 Superficial
 Subcutaneous
 Systemic
 Opportunistic
 Superficial Mycoses
infections of skin, hair, nails, and mucosa

• 4 Infections
1.Tinea versicolor
2.Tinea nigra
3. Piedra
4.Dermatophytosis
Epidermophyton, Microsporum, Trichophyton
Dermatophytes - secrete keratinase
Person to person, animal to person
Direct contact with infected hair, skin
Tinea capitis (scalp, hair), Tinea pedis (athlete’s
foot), Tinea unguium (nails)
 With
 Subcutaneous Mycoses
 Subcutaneous mycoses (mycetoma) - are fungal
infections beneath the skin
 Examples of subcutaneous mycoses include
 Chromomycosis
 Maduramycosis
 Sporotrichosis
Spores or hyphae planted directly into puncture
wound
 Infection not contagious
 Systemic Mycosis
 Deep, involves tissues and organs
 Saprophytic fungi, live in soil
 Inhalation of spores - in the lung, spreads
other tissues
 Often resemble tuberculosis
 Not contagious
 Blastomycosis - Blastomyces, flu-like,
pulmonary
 Coccidioidomycosis - Coccidiodes, San Joaquin
Valley Fever
 Histoplasmosis - Histoplasma, pulmonary,
Ohio Valley Fever.
 Paracoccidioidomycosis- Paracoccidiodes
 Opportunistic Mycoses
 Harmless in normal habitat (includes normal flora),
pathogenic in debilitated host
 Mucormycosis – caused by Rhizopus or Mucor in patients
with ketoacidosis
 Cryptpcoccosis-caused byCryptococcus neoformans
 Aspergillosis – caused after inhalation of Aspergillus
spores in those with lung disease or cancer
 Candidiasis – caused by Candida albicans
 Vulvovaginal following antibiotic therapy
 Thrush of mouth and throat
 LABORATORY DIAGNOSIS
 Specimens:
• Skin scrapings, nail clippings, hairs
• Scrapings from mucous membrane
• Scrapings, crusts, aspirated pus, tissue biopsy.
• Blood, CSF etc in systemic mycoses.
 Microscopy:
• KOH mount – KOH dissolves keratin and cellular
material but does not affect fungi. The specimen is
placed on a slide, and a drop of 10-20% KOH is added
and covered with a coverslip.
• KOH dissolves keratin and other cells and clears
debris making the fungal elements more clear in 5-20
min depending on the nature of the specimen. Then
examined microscopically.
KOH preparation
 LACTOPHENOL COTTON BLUE

• The lactophenol cotton blue (LPCB) wet mount

preparation is the most widely used method of
staining and observing fungi and is simple to prepare.
The preparation has three components: phenol, which
will kill any live organisms; lactic acid which preserves
fungal structures, and cotton blue which stains the
chitin in the fungal cell walls.
• Stains: gram stain, , periodic acid schiff stain
(PAS), methenamine sliver stain, giemsa stain etc.

• Direct immunofluorescence test

• Histology

• Antigen detection tests eg cryptococcal antigen in

CSF.
 Culture:
• Sabouraud’s dextrose agar is commonly used for
fungal culture.
• pH =5.6 does not allow bacterial growth.

• Drugs like chloramphenicol, cyclohexamide and other

antibiotics are added to prevent bacterial or
saprophytic fungal infection.

• Cultures are incubated at two temperatures:

• One tube at 25°C (room temperature)
• One tube at 37°C (incubator).
• This helps reveal fungal dimorphism.
• Cultures are incubated for at least 2-3
weeks and in some cases upto 6 weeks.
• Culture identification based on
microscopic and macroscopic
• Macroscopic-
1. Rate of growth- Rapid growth (< 5 days)
seen in saprophytes, yeasts, and agents
of opportunistic mycoses
Slow growth- (1-4 weeks) seen in agents
of subcutaneous and systemic mycoses
2. Pigmentation – seen on obverse and
reverse of culture media.
3. Texture – waxy, leathery, velvety,
yeast-like or cottony, or granular/
powdery
4. Colony topography- surface may be
radial grooves, folded or verrucose or
cerebriform ( brain-like)
• Microscopic-
1. Teased mount- fungal colony teased out
from the culture and the LPCB mount is
made on a slide and viewed under a
microscope. Identification is based on
the nature of hyphae ( such as septate
or aseptate, narrow or wide) and the
type of sporulation (conidia or sporangia)
2.Other methods- for Candida, Germ tube
test, sugar fermentation, and sugar
assimilation tests are done
• Immunological methods-
1. Antibody detection- ELISA, immune
diffusion test, agglutination test
2.Antigen detection- latex agglutination
test
• Molecular methods- PCR
 Useful Properties of Fungi

Source of food Antibiotic production

e.g. mushrooms e.g. Penicillin from
Penicillium notatum

Fermentation - Production of
alcohol, bread, cheese
e.g.
13.04.09 Sacchromyces spps 59
 Useful Properties of Fungi

Ergot from Claviceps Vaccines for Hepatitis B –

purpurea, used to induce Sacchromyces cerevisiae
uterine contractions

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• MULT PLE CHOICE QUESTIONS (MCQs)
1. Fungi which do not have sexual stage
(a) Zygomycota
(b)Ascomycota
(c) Basidiomycota
(d)Fungi imperfecti
2. Microscopically diagnosed by
(a) Albert stain
(b)Negative stain
(c) KOH mount
(d)ZN stain
 Suggested Books
• Textbook of Microbiology by
Ananthanarayan and Paniker
•Essential Microbiology for Dentistry
Textbook by Lakshman P. Samaranayake
•A Textbook of Microbiology for Dental
Students by D. R. Arora
•Textbook of Microbiology for Dental
Students by Surinder Kumar
•Textbook of Medical Microbiology by
Apurba Sankar Sastry