Jimma University

Faculty of Health Sciences

School of Medical Laboratory Sciences
Introduction to Medical Mycology
1

By: Mengistu A (MSc, Asst. professor of Medical Microbiology)

 Objectives (summary)
2 ❑ Upon completion of this section, you will be able
to:
What is Mycology and what is medical Mycology?

List the differentiating characteristics of fungi from

bacteria.

Explain the morphology of medically-important fungi

Describe the classification of fungi

Reproductions of fungi

Discuss diagnosis of medically-important fungi

 Objectives (summary)
3 ❑ Upon completion of this section, you will be able to:
What is Mycology/medical mycology?

List the differentiating characteristics of fungi

Explain the morphology of medically-important fungi

Describe the classification of fungi

Discuss fungal ecology and metabolism

Discuss the mechanisms and disease caused by fungi

Discuss lab diagnosis, therapy, prevention and control of

medically-important fungi infections
 Introduction
❑ What is Medical Mycology ?: is the study of fungi that

infect humans
What is fungi ?
▪ Fungi are eukaryotic- possess membrane-bound nuclei

and a range of membrane-bounded organelles (e.g.

mitochondria, vacuoles, endoplasmic reticulum).

▪ Natural habitats: Water, soil and decaying organic debris

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1.1.General characteristics of fungi
❑ Fungi are plant structures, but lack the definite root, stem,
and leaves.

❑ Their cell walls are mainly composed of polysaccharides

like chitin and mannan as well as glycoprotein and lipids.

❑ Cell membrane contain Ergosterol (sterols) rather than

cholesterol.

❑ Fungi are non motile

❑ They divide asexually, sexually or by both process.
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General characteristics……..
❑ Fungi are non-photosynthetic organisms
▪ Do not contain chlorophyll pigments and are incapable
of photosynthesis
▪ Are unable to manufacture their own nutrients,
▪ They require external source of preformed organic
compounds as a source of carbon; hence they are
frequently associated with decaying matter to
obtained their nutrients
▪ But some fungi use living tissue as a food source e.g.
Candida (yeast)
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General characteristics……….
❑ Fungi grow as yeasts, molds or a combination of both (i.e.
dimorphism).
❑ Most fungi are filamentous- Composed of individual
microscopic filaments called hyphae, which exhibit apical
growth and which branch to form a network of hyphae
called a mycelium
❑ All fungi are eukaryotic (with true nucleus).
❑ Most fungi are found as saprophytic in the soil.

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General characteristics……..
❑ Most of the fungi are Multicellular, but may be unicellular

❖ Unicellular Fungi→ Microscopic fungi exist as either

molds that grow on bread, fruits and cheese or Yeasts
used in baking industry e.g., Yeast (Candida), Cryptococcus,
etc.

❖ Multicellular → Macroscopic fungi often called fleshy

fungi and are filamentous organisms e.g., Aspergillus,
Trychophytan, Microsporum, Epidermophytan etc.
Characteristics of fungi ……..

❑ They are heterotrophic and obligate aerobes and some are

facultative anaerobes; but none are obligate anaerobes.

❑ Fungi are mainly pathogens of plant but relatively few

are pathogens of human.

❑ Cause serious diseases in immuno-compromised individuals.

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Comparison of fungi and bacteria

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1.2. Morphological of fungi
❑ Most medically important fungi are exist into two
morphological forms:
1. Filamentous fungi, also referred to as moulds
2. Yeasts
3. Dimorphic fungi (having yeast and filamentous forms)
❑ Moulds: multi-cellular, filamentous forms of fungi mostly
found outside the body and at room temperature.

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Moulds…………
❑ Moulds are grown as branching filaments called
hyphae.
❑ Hyphae is the basic microscopic unit of the molds

❖ It is long tube like structure responsible for the fluffy

or cotton ball appearance of the macroscopic mold
colony.

❑ Hyphae grow as an intertwined mass of filaments

collectively called mycelium

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Mold…………cont’d
❑ Mold are classified and identified partially on the basis
of whether the hyphae are septate or aseptate
▪ Septate hyphae: filaments with cross-walls or septa that
partition the hyphae in to individual cellular compartments

▪ Aseptate hyphae: - Hyphae without septa are referred as

aseptate; no physical boundaries to distinguish individual
cells in the hyphae (feature of zygomycetes).

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Hyphae cont’d

Fig . Septate and Aseptate hyphae

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Moulds…………
• Moulds of medical importance
– Dermatophytes: Epidermophyton, Microsporum & Trichophyton
– Fungi causing Mycetoma
– Fungi causing Chromomycoses
– Aspergillus species
– Penicillium marneffei
– Zygomycetes: Mucor, Absidia, Rhizopus, Basidiobolus species

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Mold (Hyphae)
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F I G U R E: Mold forms. The tube-like hyphae constitute their basic structure.
Examples of spores and conidia and of the structures that bear them are
shown. They develop from the hyphal wall.
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2. Yeasts
❑ Yeasts: are single cells (unicellular fungi), usually spherical
to round in shape and occurs singly, in pair or form a chain.
❑ Structure mostly found inside tissue or at body temperature
❑ Reproduce by an asexual reproduction called budding.
❑ A daughter cell called a blastoconidia (often called a
blastospore) is formed on the surface of the parent cell.
❑ Form psedohyphae not a true hyphae
❑ Form creamy mucoid colonies on culture media.
Ex: Cryptococcus neoformans, Candida albicans ,Trichosporon beigelii

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 2. Yeasts………..
❑ Some yeasts grow partly as yeast and partly as chains of
elongated budding cells to form pseudohyphae.
Ex: Candida albicans

2. Yeasts

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Fig . yeast and its pseudohphae
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3.Dimorphic fungi:
❑ Dimorphic fungi exist as yeast in the host tissue and in the cultures
at 370c and exist as mold forms in the soil and in the culture at RT
(22- 250c).
❖ E.g., Blastomyces dermatitidis, Paracoccidioides brasiliensis,
Histoplasma capsulatum, Sporothrix schenckii & Coccidioides
immitis are examples of medical importance dimorphic fungi.
❑ Most of them known to cause systemic infections, especially,
more severe infections in immunocompromised patients.

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1.3. Fungal Reproduction
❑ Fungi produce by sporulation (process of making
spores) and spread through the environment.

❑ Spores can be generated either asexually or sexually

❑ Many pathogenic fungi are ‘imperfect’, i.e. they are

known only to produce vegetative non-sexual spores and
conidia.

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Fungal reproduction…….. cont’d
❑ Fungi that exhibit sexual phase reproduction are known
as the perfect fungi

❑ Fungi without sexual cycle of reproduction are known as

imperfect.

❑ Except for the deuteromycetes, all fungi produce sexual

spores
❑ Asexual spores that are generally responsible for initiating
human infection
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 Fungal Reproduction…….
❑ Some important asexual conidia
▪ Arthrospores (Arthroconidia)– result from the fragmentation of
hyphal cells into rectangular thick walled spores. e.g. Microsporum

▪ Chlamydospores(Chlamydoconidia)–thick walled resting spores

developed by rounding up and thickening of hyphal segments.

e.g. Epidermophyta
▪ Blastospores (Blastoconidia)– formed by budding from parent
cell e.g. Candida by budding
• Sporangiospores (Sporangioconidia)– spores are formed within the
sporangium. They develop on the ends of hyphae called sporangiospores.
(Sporangium is a round, hard case that holds thousands of spores).Ex: Mucor and
Rhizopus.

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Types of sexual spores
❑ Basidiospores
– Mushroom is a common fungal fruiting body that produces
basidiospores,

– Spores formed at the tip of a club like structure called a basiduim.

❑ Ascospore
– Ascospore is found in a sac-like structure called an ascus (plural asci)
– Several asci are usually contained in a single large fruiting body.
❑ Zygospores
– are encased in a thick, darkly pigmented wall - makes resistant to
extremes in temperature and desiccation.

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1.4 . Classification of fungi
❑ Classification of fungi is based on:
– Spore morphology
– Septation of hyphae
– Mode of reproduction
❑ There are four categories of fungi
– Ascomycetes
– Basidiomycetes
– Deuteromycetes
– Zygomycetes
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Classification of…..

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Ascomycetes
❑ Includes yeasts and many of the common molds that can
produce ascospores
❑ Have septate hyphae
❑ Reproduce by both sexual (ascospores) found within a sac or
ascus and asexual (conidia) methods

❑ Include many fungi beneficial to man, such as the yeasts of the

genus saccharomyces that leaven bread and ferment alcoholic
beverages.

Example: Some Aspergillus and Pencillum spps.

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Basidiomycetes
❑ Consists of higher fungi that produce exogenous sexual
basidiospores on a basiduim and have septate hyphae.

❑ Mostly are saprophytic organisms and are rarely

encountered clinically.
❑ Also reproduce by asexual (conidia) methods
❑ This class includes mushrooms, toad stools, and puffballs
are considered fleshy fungi
❑ The only human pathogen is Cryptococcus neoformans.

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Deuteromycetes (imperfect Fungi)
❑ Have both yeasts and mould forms.
❑ Don’t known sexual cycle of reproduction and hence called
fungi imperfect
❑ Many of pathogenic fungi are included in this class.
❑ They have septate hyphae and produce conidia from
conidiophores and conidiogenous cells.

❑ The yeasts reproduce by budding, and the moulds produce

conidia.
Example: Candida species and Epidermophyton species
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Zygomycetes
❑ This group, having a worldwide distribution, may be found
in soil or decaying vegetable matter.
❑ They may also be found on stored grain, fruits and
vegetables, compost, and air.
❑ They are usually harmless to humans, but may be
allergenic and are known to cause opportunistic infections.
❑ They are characterized by aseptate hyphae, asexual
sporangiospores, and sexual zygospores.
Example: Rhizopus nigricans (black bread mold) and Mucor -
common soil fungi.

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1.5. Fungal Ecology and metabolism
▪ All fungi are carbon heterotrophs, which means they are
dependent on exogenous nutrient substrates as sources of
organic carbon

▪ With a few exceptions, fungi are obligate aerobes.

▪ The known metabolic types of fungi include thermophilic,

psychrophilic, acidophilic, and halophilic species.

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Ecology ………cont’d
❑ The metabolic capabilities of fungi are exploited in the food
industry (e.g., in the production of bread, wine, beer, cheese, or
proteins) and in the pharmaceutical industry (e.g., in the
production of antibiotic substances, enzymes, citric acid, etc.).
❑ The metabolic activity of fungi can also be a damaging factor.
▪ Fungal infestation can destroy foods, wooden structures,
textiles, etc.
▪ Fungi also cause numerous plant diseases, in particular
diseases of crops.

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Ecology ………cont’d
▪ In general the metabolism of fungi can be;
I. Saprophytic fungi:
• Most fungi are saprophytic, free living organisms obtain
their nutrients from dead organic matter

• Important decomposers in biogeochemical cycles.

II. Mutualistic fungi
• Many fungi have evolved successful relationships with
plants or microbes in which both partners benefit E.g.
Lichen is a composite of a fungus and a photosynthetic microbe,
either an algae or cyanobacterium.
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Ecology ………cont’d
III. Parasitic fungi
▪ Some fungi acquire nutrients by attacking live animals or
plants.

▪ Of the 150,000 known fungi species, about 50 have been

identified as primary human pathogens.

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 Chapter-two
Laboratory Diagnosis of Fungal infections

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2.1. Mycological specimen

▪ The diagnosis of fungal infection depends entirely on the

selection and collection of an appropriate clinical specimen.

▪ Microscopy and culture are the main ways in which fungal

infections are diagnosed in the laboratory.

• In addition, immunological methods that help in the diagnosis of

fungal infection.

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Mycological specimen……..

❑ Specimens for fungal microscopy and culture are transported to

the laboratory in a sterile containers.
❑ Specimens used for diagnosis of fungal infection include:
• Skin, nail, hair scrapings and swabs
• Respiratory tract secretions
• Cerebrospinal fluid (CSF)
• Blood and bone marrow aspirates
• Tissue biopsies from visceral organs
• Urine and urogenital specimen

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2.2. Examination methods
• Direct microscopy after KOH preparation using 10%
KOH

• DNA probe test

• Serologic tests- based on Ag and Ab reaction.

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Examination methods …..

I. Direct microscopy:
▪ Direct Microscopic method includes:
• Saline wet mount preparation

• Potassium hydroxide (KOH) preparation

• Staining methods

▪ Note : Microscopic methods should be confirmed with

cultures or antigen testing, when available.

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Examination methods …..

A. KOH preparation: are used in the initial examination of

keratinized tissue suspected of fungal infection. e.g.
dermatophytes (ringworm fungi) in skin scrapings, nails or hair.
Principle
– KOH (20%w/v) dissolves keratin in skin, hair or nail specimens,
facilitating the observation of the organism’s morphology.

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KOH preparation…….
Procedure
1. Put one drop of KOH reagent on slide, place a small portion of
the material (skin scrapings, hair, and nail) to be examined.
2. Press cover slip down on sample.
3. Warm the slide gently to dissolve keratinized cells. Do not boil.

4. Allow specimen to clear, approximately 20 minutes.

5. Examine under low and high- dry magnification.

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Examination methods …..
• Growth and isolation
– Fungi grow easily but slowly in their appropriate culture
media.

– The most commonly used media is Sabouraud’s Dextrose

agar (contains glucose and beef extract).

• It inhibits bacterial growth.

– Most fungi grow as yeast at 37oC and as mold at 25oC

(they are dimorphic).

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 Chapter-II
Disease mechanism and Diseases
caused by Fungi

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3.1. Disease mechanism of fungi
❑ Little is known about the pathogenic mechanisms and virulence
factors involved in fungal infections.
Adherence
• Some fungus like yeasts, are able to colonize the mucosal
surfaces of the GI and female genital tracts.

• Adherence usually requires a surface adhesion on the microbe

and a receptor on the epithelial cell.
• In the case of C. albicans, mannoprotein components extending from the cell
wall have been implicated as the adhesin, and fibronectin a receptor

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Disease mechanism……. cont’d
Invasion
▪ Passing an initial surface barrier, whether skin, mucous
membrane, or respiratory epithelium, is an important step
for most successful pathogens.

▪ Fungi that initially infect the lung must produce conidia

small enough to be inhaled past the upper airway defenses
( e.g. arthroconidia of Coccidioides immitis)

▪ Invasion across mucosal barriers may involve enzymes

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Disease mechanism……. cont’d
❑ Triggered by temperature and possibly other cues, dimorphic
fungi change their morphology and growth to a more invasive
form.

❑ Extracellular enzymes (e.g., proteases, elastases) are

associated with the hyphal form of Candida and with the
invasive forms of many of the dimorphic and other pathogenic
fungi.

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Disease mechanism……. cont’d
Tissue Injury
▪ None of the extracellular products of opportunistic fungi or
dimorphic pathogens have been shown to injure the host
directly during infection in a manner analogous to bacterial
toxins.

▪ A number of fungi do produce exotoxins, called

mycotoxins, in the environment but not in vivo.
▪ Mycotoxcosis may result from ingestion of fungal
contaminated foods. Example – Poisonous mushroom

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Disease mechanism……. cont’d

❑ The injury caused by fungal infections seems to be due

primarily to

• The inflammatory and immunologic responses that are

stimulated by the prolonged presence of the fungus.

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3.2.Predisposing factors for fungal diseases

▪ Age : Young and elderly are more susceptible

▪ Immune compromise : AIDS, Diabetes mellites, Use of

corticosteroids, organ transplantation
▪ Trauma or injury : subcutaneous mycoses
▪ Contact with contaminated soil / animal / bird
excreta / infected persons

▪ Inhalation of contaminated air with fungal spores.

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3.3. Diseases caused by Fungi
❑ Few species of fungi cause diseases in human.
❑ Diseases caused by fungi are called mycoses and it
can be;
– Superficial mycoses- Localized, e.g., hair shafts
– Cutaneous mycoses- Affect hair, skin, and nails
– Subcutaneous mycoses-Beneath the skin
– Systemic (Deep) mycoses- Deep within body
– Opportunistic mycoses- Fungal infection that occur in
immune compromised individuals like during HIV. e.g.
Candidiasis, Aspergillosis etc.

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