INTRODUCTION TO MYCOLOGY

Dr. Chandrashekhar Unakal

Lecturer In Medical Microbiology
FMS; UWI
 Introduction / Structure
Mycology is the study of fungi – Yeast and Mold
Yeast are unicellular , nucleated rounded fungi while molds are
multicellular , filamentous fungi
Yeast reproduce by a process called budding while molds produce
spores to reproduce
The name Fungi derived from mykos - meaning mushroom .
The Fungi possess rigid cell walls containing :- Chitin ,
Glucan , Mannans and Cell membranes contains : Ergosterol
All fungi are eukaryotic organisms – nucleus & nuclear membrane ,
endoplasmic reticulum , mitochondria .
 Introduction Conti….

 Yeasts and Molds have different structural and

reproductive characteristics
 Fungi are important as pathogens of Plants , Human and
Animals .
 Over 70% of all Plant diseases are caused by fungi .
I. FUNGI (Mycology)

 Many are ecologically important saprophytes ( Consume

dead and decaying matter ) ; Others are parasites .
 Most are multicellular , but yeasts are unicellular .
 Most are aerobes or facultative anaerobes .
 Cell walls are made up of chitin ( Polysaccharide ) .
 Most human fungal infections are nosocomial and / or occur
in immunocompromised individuals ( Opportunistic
infections ) .
 CHARACTERISTICS OF FUNGI
Morphological Classification Of Medically Important
Fungi
 1. Yeasts
 2. Yeast like fingi
 3. Mold / Filamentous fungi
 4. Dimorphic fungi
1. Yeasts
 Unicellular fungi , non filamentous , typically oval or
spherical cells ( Gram +ve Yeast ) . Reproduce by budding
 E .g : Creptococcus neoformans :- A true yeast fungus ,
capsulated & which is demonstrated by Indian ink staining .
 On culture on SDA – produce smooth , creamy colony .
 2 .Yeast like fungi
 Grow partly as yeast & partly as elongated cells resembling hyphae which is
called Pseudohypha . Budding yeasts can form pseudohypha , a short chain of
undetached cells . Reproduce by Budding .
 E g : Candida albicans :- A yeast like fungus with pseudomycellium and Germ
tube test positive .
 Culture on Sabouraud’s dextrose agar ( SDA ) - shows creamy white colony .
Gram stain shows – Gram positive budding yeasts .
 3 .Molds / Filamentous Fungi :
 Multicellular , filamentous fungi . Form true mycelia & reproduce by
formation of different types of spore .
 Identified by physical appearance , colony characteristics , and
reproductive spores.E.g : Aspergillus spp , Penicillium spp ,
Dermatophytes ect .
 ◦Thallus : Body of a mold or fleshy fungus . Consists of many hyphae .
 ◦Hyphae ( Sing: Hypha ) : Long filaments of cells joined together .
 ◦Septate hyphae : Cells are divided by cross-walls ( septa ) .
 Aseptate hyphae : Long continuous cells that are not divided by septa .
 Mycelium : Large , visible , filamentous mass made up of many hyphae .
Penicillium spp .
 Brush / Broom like arrangement of Conidia .
 Colonies initially Velvety and White but Later become
Powdery & Blue Green .

Colony on SDA
Lactophenol Cotton Blue stain
 Aspergillus spp. Lactophenol Cotton Blue stain

Conidiophore , Sterigmata ,
Conidia seen

SDA ( Sabourauds ) /PFA

(Potato Flake Agar) -Green
coloured colonies of
Aspergillus fumigatus
 4 . Dimorphic Fungi
 Can exist as both multicellular fungi ( molds ) and yeasts depends on condition of
growth . Mold form produces aerial and vegetative hyphae. Yeast form reproduces by
budding . Primary site of infection – pulmonary

 Dimorphism in pathogenic fungi typically depends on temperature : At 37oC , host

tissue - Yeast form And At 25oC , soil - Mold form . E.g. : Blastomyces dermatitides
, Coccidiodes immitis , Histoplasma capsulatum , Sporothrix schenckii , Penicillium
marneffei , Paracoccidioides brasiliensis .
 Reproduction and Life cycle of fungi
 Yeast & Yeast like fungi reproduce by budding .
 Filamentous fungi can reproduce asexually by fragmentation of their hyphae .
 Fungal spores are formed from aerial hyphae and are used for both sexual
and asexual reproduction .
 Asexual spores : Formed by the aerial hyphae of one organism . New
organisms are identical to parent .
 Conidiospore : Unicellular or multicellular spore that is not enclosed in a sac .
 Chlamydospore : Thick-walled spore formed within a hyphal segment .
 Sporangiospore : Asexual spore formed within a sac ( sporangium ) .
 Sexual spores : Formed by the fusion of nuclei from two opposite mating
strains of the same species . New organisms are different from both parents .
 DIVISIONS OF FUNGI – Systematic
Classification – Fungi ( Mycetae ) .
 1. Deuteromycota [ Deuteromycetes ]
( Fungi Imperfecti )
 Not known to produce sexual spores .
 Reproduce asexually .
 Catch - all category for unclassified fungi :

 Candida albicans : Causes yeast infections of vagina in

women . Opportunistic infections of mucous membranes in
AIDS patients .
 Malassezia furfur ( Tinea versicular ) , Sporothrix schenckii
( Rose gardeners ) .
 2. Zygomycota [ Zygomycetes ]-
(Conjugation Fungi)

 Also known as bread molds .

 Saprophytic molds with Aseptate hyphae .
 Asexual Reproduction : Occurs via Sporangia .
 Sporangiospore : Asexual spore enclosed within a
sporangium or sac at the end on an aerial hypha
 Sexual Reproduction :Occurs through conjugation &
result in a Zygospore .
 Zygospores : Sexual spores which are enclosed in a
thick , resistant wall. E.g: Rhizopus –Opportunistic
infection in diabetes patients , Mucor .
 3. Ascomycota [ Ascomycetes ]- (Sac
Fungi)
 Molds with septate hyphae and some yeasts .
 Asexual Reproduction : Via Conidia - Conidiospores not
enclosed in a sac. Become airborne easily. Form chains (
Broom -like structures ) .
 Sexual Reproduction : Producing Ascospores enclosed in
a sac-like structure ( Ascus ) .
 Include common antibiotic producing fungi and yeasts ,
and several human pathogens .
 ◦Penicillium notatum ( Produces penicillin )
 ◦Aspergillus ( Carcinogenic aflatoxin in peanuts )
 ◦Blastomyces ( Respiratory infections )
 ◦Histoplasma capsulatum ( Respiratory and systemic
infections )
 4. Basidiomycota [ Basidomycetes ] - ( Club Fungi )

 Have septate hyphae .

 Include Mushrooms - Amanita , Cryptococcosis – Cryptococcus
neoformans
 Sexual Reproduction :- Produce basidiospores : Spores formed
externally on a club shaped sexual structure or base called basidium.
 Asexual Reproduction :-
 Through budding : e.g. Cryptococcus neoformans - Causes
opportunistic respiratory and CNS infections in AIDS patients .
 ◦ Amanita spp : Mushroom produces lethal toxins to humans .
FUNGAL INFECTIONS ( Mycoses )

 Fungal infections or diseases are referred to as Mycosis .

 In general , humans have a high level of innate immunity to fungi and most of the
infections they cause are mild or self limiting . This resistance is due to :
 Fatty acid content of skin ; pH of the skin ; mucosal surfaces ; cilia of respiratory
tract . When fungi do pass the resistance barrier of the human body & establish
infection .
 The Fungal Infections are Classified according to the tissue levels colonised &
Include following types :-
 1. Superficial mycoses
 2. Cutaneous / Dermatophytes Mycoses
 3. Sub-cutaneous Mycoses
 4. Systemic Mycoses
 5. Opportunistic Mycoses
 Tinea versicolor-Pitriasis versicolor
1 . Superficial mycosis Tinea nigra
White piedra
Black piedra

Disease Fungus manifestation

Tinea versicolor Malassezia furfur Decolourisation of skin .

Tinea nigra Exophiala wereneckii Macular patches on skin.( Palm

& Soles ) .

White piedra Tricosporum beigelii Nodules on hair shaft of axilla ,

pubic .

Black piedra Piedraia hortae Black nodules on hair shaft of

scalp , beard .
Tinea versicolor Tinea nigra White piedra Black piedra

 caused by Malassezia Characterised by soft , white or light Black hard nodules on hair shafts of
Dark-colored macule due to accumulation
furfur of a melanin like substance in the brown nodules , on axillary / pubic scalp & beard .Caused by Piedraia
Hypopigmentation fungus.Caused by Exophiala wereneckii hair.Caused by Tricosporum beigelii hortae .
 2 . Cutaneous mycoses:
 Fungal infections of the skin , hair , and nails ( Dermatophytes , dermatophytoses )

 Infection is transmitted by direct contact or contact with infected hair ( hair salon ) or cells ( nail
files ) . Examples : - Ringworm ( Tinea capitis and T. corporis ) ; Athlete’s foot ( Tinea pedis ) ;
Jock itch ( Tinea cruris )

Ringworm skin infection: Severe nail infection with Candida albicans

Tinea corporis Trichophyton rubrum infection of the nails.
 Dermatophytes – Macroconidia :
Trichophyton rubrum Microsporum gypseum Epidermophyton floccosum

Pencil shaped - Septate Spindle shaped - Club shaped in clusters -

Up to 10 , Thin walled . Thick walled - 4-6 septa . Smooth walled - 3-4 septa .
 Ring worm

Tinea pedis

Tinea capitis Tinea corporis

Tinea manuum&T.
pedis&T.unguium
 3 . Subcutaneous mycoses
 Fungal infections beneath the skin , involving the dermis ,
subcutaneous tissues , muscle and fascia , also the bones .
 Caused by saprophytic fungi that live in soil or on vegetation .
 Infection occurs by implantation of spores or mycelial
fragments into a skin wound .
 Can spread to lymph vessels . They include Sporotrichosis
( Sporothrix schenckii ) ; Chromoblastomycosis ( Fonsecea
pedrosoi , Fonsecaea compactum ) ; Mycetoma ( Madurella
grisea , Madurella mycetomatis ) .
Sporothrix schenckii

Lymphocutaneous Sprotrichosis
 Sporotrichosis ( Rose-
Gardener’s Disease ).. Portal of
entry – Skin .

 Lymphocutaneous variety
occurs when contaminated
plant matter penetrates the skin
& the pathogen forms a nodule,
then spreads to nearby lymph
nodes .
Primary sore and series of secondary
nodules along lymphatic chain in the arm
 4 . Systematic Mycoses
Fungal infections deep within the body . Can affect a number of tissues & organs
- causing pulmonary & systematic infection from airborne conidia .
a) Blastomycoses : ( Blastomyces dermatitidis ) - Acute & chronic
infection of lungs common in USA & Canada .
b) Histoplasmoses : ( Histoplasma capsulatum ) :- Associated with
droppings of bats & birds . Human infection by inhalation of spores -
Initial infection in Lungs . Later spreads through blood to most
Organs . Common in USA .
c) Coccidioidomycoses : ( Coccidioides immitis ) - Causes pulmonary
infection ( Resembles Tuberculosis ) common in USA and Mexico .
d) Cryptococcosis : ( Capsulated yeast - Cryptococcus neoformans )
–Found in droppings of birds , infection frequently seen in patents
with AIDS with meningitis & lung infections .
Opportunistic Mycoses
Infection under special condition such as Immuno- compromised
patient , associated with HIV , wide use of antibiotics , steroids .

a) Candidiasis : ( Candida albicans )- Yeast like , causes oral thrush ,

cutaneous candidiasis , meningitis , septicemia & nail infection .
b) Aspergillosis : ( Aspergillus sp. ) – Filamentous fungi , causes
pneumonia , aspergilloma, otitis , keratitis & aspergillous asthma.
c) Mucormycoses / zygomycosis : ( Mucor , Absidia , Rhizopus ) –
Cotton like growth on roof of mouth , paranasal sinuses .
d) Pneumocystitis: ( Pnumocystic carinii )- Causes pneumonia with
high mortality in AIDS , malignancies & autoimmune disease .
 TOXICOSIS : 2 types of fungul toxicosis

 A – Mycotoxicosis : Result from consumption of feeds or food

products contaminated with toxin - producing fungi e .g : Afla- toxin
by Aspergillus flavus ( grow in ground nuts & cereals ) ; Mycotoxin by
Fusarium spp ; Fumigatin toxin by Aspergillus fumigatus .

 B – Mycetismus : Results from ingestion of fungi containing

preformed toxin e. g : Mushroom poisoning .
 Laboratory Diagnosis of fungi

 Specimens for fungi identification in the microbiology laboratory

could include :
 Hair ; Nail ; Skin scrappings
 Tissue biopsy
 Cerebrospinal fluid ( CSF )
 Sputum , Broncho-alveolar larvage ( BAL )
 Blood or Any other body fluid : – Urine , Pleural fluid ,
Peritoneal fluid
Lab Diagnosis-Microscopy & Staining

Microscopy – Staining types

 Wet preparations , treat with 10% KOH , with 5% glycerin
or Calcoflour staining .
 Lactophenol cotton blue staining
 Haematoxylin & Eosine stain
 India Ink stain
 Woods lamp :– Fluorescence of fungi under UV light for
Microsporum spp .
 Fungal Culture

Culture –Growth slow . ( Tube & Slide culture ) .

 Sabourauds’s Dextrose agar ( SDA ) – favours fungal growth because of
low pH , Potato dextrose or rice flake or cornmeal agar at 25 or 37oC
 Mycosel agar – selective for pathogenic fungi because of chloramphenicol
and cycloheximide in medium
 Growth On the agar – Visualize morphology of colony , Spores , Cells ,
Hypha ( aseptate , septate ) .
 Spores – conidiospores , sporangiospore , arthrospore , chlamydospore
 Yeast morphology – size , pseudohyphae , capsule ( India Ink Stain )
 Biochemical tests
 Germ tube formation
 Pseudohyphae formation
 Urease ; Fermentation and Assimilation studies

 Immunoassay
 Latex agglutination test , Enzyme assay ( ELISA ) .

 Molecular tests
 Polymerase Chain Reaction ( PCR ) .
 Treatment of Fungal Infections
 Macrolides e.g.Amphotericin B , Nystatin : Bind to Ergosterol
 Azoles-e.g. Ketoconazole , Econazole , Fluconazole , Miconazole
Ketoconazole : Termination of cell membrane synthesis .
 Allylamines –e.g. Terbinafine : Inhibits Ergosterol synthesis .
 Pyrimidine analogs – Flucytosine :Inhibits DNA & RNA synthesis
 Grisans –Griseofulvin :Inhibits Fungal Mitosis.Rx -Dermatophytosis
 Caspofungin : Inhibits fungal cell wall synthesis .
Thank You.
 Sample spotter Question MDSC 1002
Q1.Thefollowing picture shows culture on SDA and Lactophenol cotton blue stain of
fungus. A fungi characterized by a vesicle – swollen tip to the conidiophore stalk upon
which phialides are mounted .

phialides

Growth on SDA
vesicle

Which of the following is a disease caused by the fungi shown above in immuno-
compromised individuals ?
A. Onychomycosis
B. Otomycosis
C. Aspergillosis
D. Penicilliosis