Maseno University

School of Public Health and

Community Development
Department of Public Health
PHT 222:Medical Microbiology
MYCOLOGY 1
 Mycology
• Is the study of fungi
• Fungi are eukaryotic organisms belonging to the
Kingdom Fungi
• They are ubiquitous in nature
• Comprise of 250,000 species of which 150 cause
diseases in humans
• Fungi are responsible for fungal diseases called
mycoses;
• Superficial mycoses/cutaneous
• Subcutaneous mycoses
• Systemic mycoses
• Opportunistic mycoses-infections due to fungi with low
inherent virulence 2
 Mycoses
• Superficial mycoses: affecting the skin, hair or nails, e.g.
dermatophytosis (ringworm) and pityriasis versicolor. They
are confined to the body surfaces and do not directly
involve living tissues.
• Subcutaneous mycoses: which are referred to as mycoses
of implantation, e.g. mycetoma, chromomycosis,
subcutaneous phycomycosis, rhinosporidiosis, and
sporotrichosis. They are acquired when the pathogen is
inoculated through the skin by minor cuts or scratches or by
thorn or splinter woods.
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 Mycoses cont…..
• Systemic mycoses: which are often referred to as
deep mycoses, e.g. histoplasmosis, blastomycosis,
paracoccidioidomycosis, aspergillosis, and
coccidioidomycosis. They are acquired by
inhalation and may spread from the lung and
involve any part of the body. Widespread infections
can be fatal. Skin lesions are often present.

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•Fungi differ from bacteria in several aspects

Difference between bacteria and

fungi
FEATURE FUNGI BACTERIA
Nucleus Eukaryotic Prokaryotic
Cytoplasm Mitochondria, ER present Both absent
Cell membrane Sterols Not there except
mycoplasma
Cell wall content Chitin Peptidoglycan
Spores For reproduction For survival
Thermal dimorphism Yes(some) No
Metabolism Require organic carbon Many don’t require organic
carbon

No obligate aerobe Many obligate aerobes

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 General properties of fungi
1. They are eukaryotic; cells contain membrane bound cell organelles
including nuclei, mitochondria, golgi apparatus, endoplasmic reticulum,
lysosomes etc. They also exhibit mitosis.
2. Have ergosterols in their membranes and possesses 80S ribosomes.
3. Have a rigid cell wall made of chitin.
4. Are chemoheterotrophs (require organic compounds for both carbon and
energy sources) and lack chlorophyll and are therefore not autotrophic.
5. They obtain nutrients as saprophytes (live off of decaying matter) or as
parasites (live off of living matter).
6. Most are obligate aerobes and some are facultative anaerobes there are no
obligate anaerobes.
7. Typically reproduce asexually and/or sexually by producing spores. Fungi
with known sexual stages are called perfect fungi, and those without the
sexual stages are known as imperfect fungi.
8. They grow either reproductively by budding or non-reproductively by
hyphal tip elongation.
9. Food storage is generally in the form of lipids and glycogen.

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• Are calssified based on;
Classification
– Morphology - exsist in three forms Moulds, Yeasts, Dimorphic
– Reproduction - spore formation.
– Tissues they infect - subcutaneous, cutaneous, systemic
– Scientific classification - fungal species is assigned to a phylum as well as the
appropriate Class, Order, and Family.
– Medically important fungi are in four classes;
• Ascomycetes - produce ascospores, have a spetate hyphae e.g Blastomyces
spp, Histoplasma spp, Microsporum spp, Trichophyton spp, Coccidioides
spp, and Candida spp.
• Basidiomycetes - produce basidiospores e.g mushrooms, Cryptococcus
neoformans.
• Deuteromycetes - fungi with no sexual spores e.g Penicillium spp. and
Aspergillus spp.
• Zygomycetes - form zygospores and are non-septated e.g mucor and
rhizopus

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 Cont
• The largest phylum is the Ascomycota (or
ascomycetes), which includes more than 60%
of the known fungi and about 85% of the
human pathogens. The remaining pathogenic
fungi are zygomycetes or basidiomycetes

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 Structure of fungi
• Fungi exist in three forms; moulds, yeast and dimorphic fungi
Moulds (Molds):
• These are multicellular fungi that form branching filaments called hyphae
• A mass of hyphae ( branching hyphae) collectively make up the mycelium.
• an organized body of hyphae is a fruiting body (e.g., a mushroom).
• There are two kinds of hyphae;
 non-septate (coenocytic)/aseptate- no cross walls on the septa
 Septate - hyphae are divided into cells by cross-walls called septa. Each
compartment may have a single nucleus (uninucleated) or may have more than
one nucleus (multinucleated). The hyphae of many pathogenic moulds are
septate
• Mycelia can be;
– Vegetative mycelia penetrate the medium to obtain nourishment for the
organisms e.g rhizoids;
– Reproductive mycelia produce a great variety of conidia which are borne on
specialized hyphae or conidiophores Eg: Aspergillus spp., Trichophyton rubrum
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 Yeasts:
• These are round, oval or elongate unicellular fungi,
measuring 3–15 µm.
• Single celled cells reproduces by simple budding to form
blastoconidia (often called a blastospore) is formed on the
surface of the parent cell.
• Yeast like: Similar to yeasts but produce pseudohyphae
(Elongated budding cells often linked together forming
branching chains)
• The yeast cells of Candida albicans also germinate,
forming sprout-like projections called germ tubes that
extend to form true hyphae e.g: Candida albicans.
• A few pathogenic yeasts form a capsule, forming mucoid
colonies e.g. Cryptococcus neoformans. 10
 Dimorphic fungi
• These are fungi that are able to grow as yeasts or moulds
depending on environmental conditions and temperature.
The yeast form is found in infected tissue and when the
fungus is cultured at 35–37 C and the filamentous form is
found in the soil and when the fungus is cultured at ambient
temperatures (20–30 C).
Dimorphic fungi of medical importance
• Blastomyces dermatitidis
• Paracoccidioides brasiliensis
• Histoplasma species
• Coccidioides immitis
• Sporothrix schenckii
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 Reproduction of fungi
• Reproduce both asexually and/or sexually by producing spores.
• Asexual reproduction occurs in the fungi when spores form by
mitosis. These spores can be;
– conidia/conidiospores- are small single celled microconidia or
large multicellular macroconidia formed at the tip of the hyphae
– Sporangiospores- single celled spores found within a sac called a
sporangium at the edge of a special hyphae called a
sporangiophore
– Arthrospores – single celled spores formed from fragments of
hyphae
– Chlamydospores- Single celled spores with thick walls
– Blastoconidia (blastospores): Conidial formation through a
budding process (e.g, yeasts)

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13
 Cont
• sexual reproduction- compatible nuclei unite
within the mycelium and form sexual spores.
Sexually opposite cells may unite within a single
mycelium, or different mycelia may be required.
When the cells unite, the nuclei fuse and form a
diploid nucleus. This is followed by meiosis.

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 Cont
• Sexual spores are formed less frequently
– Ascospores - single celled spores produced in a sac
called ascus. There are four to eight ascospores in each
ascus
– Basidiospores - single celled spores found in a club-
shaped structure called a basidium. Four meiospores
– Zygospores: large thick-walled spore formed when the
tips of two sexually compatible hyphae fuse

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basidiospores zygospores

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Ascopores

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 Fungal Pathogenicity (virulence
factors):
• Ability to adhere to host cells by way of cell wall
glycoproteins
• Production capsules allowing them to resist phagocytosis
• Production of a cytokines by Candida albicans that
suppress the production of complement.
• Ability to acquire iron from red blood cells as in Candida
albicans
• Ability to damage host by secreting enzymes such as
keratinase, elastase, collagenase
• Ability to resist killing by phagocytes as in dimorphic fungi
• Ability to secrete mycotoxins
• Exhibiting thermal dimorphism
• Ability to block the cell-mediated immune defences of the
host.
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 Growth and nutrition of fungi
• Most fungi grow at alow acidic pH of about 5.0, although some
species grow at lower and higher pH levels.
• Most fungi grow at about 25°C (room temperature) except for
pathogens, which grow at 37°C (body temperature).
• Fungi grow best where there is a rich supply of organic matter.
• Most fungi are saprobic (obtaining nutrients from dead organic
matter).
• They grow in high sugar concentration which inhibit other organisms
• Fungi store glycogen for their energy needs and use glucose and
maltose for immediate energy metabolism.
• Most species are aerobic, except for the fermentation yeasts that grow
in both aerobic and anaerobic environments.

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 Laboratory diagnosis
 There are five approaches to the laboratory
diagnosis of fungal diseases:
direct microscopic examination,
culture of the organism,
histology/cytology
DNA probe tests, and
serologic tests
 Specimens;
• Cutaneous mycosis; skin Scrapings, hair and nail clippings
• Swabs from mucous membranes
• Subcutaneous infection- crusts, aspirated pus, biopsies
• Systemic infections- specimen from appropriate site
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Microscopy:
– used to observe clinical specimens for the presence of fungal
elements or to identify the fungus following culture. Direct
examination of clinical specimens could be stained or unstained.
– lactophenol cotton blue stains the fungal elements blue. Lactic acid
acts as a clearing agent and aids in preserving the fungal
structures; phenol kills the organism and fixes it; glycerol prevents
drying and cotton blue provides colour to the structure.
– Wet mount: Candida (yeast cells) may be observed in urine wet
mounts
– 10-20% KOH mount: Several specimens are subjected to KOH
mount for direct examination. The material is mixed with 20%
KOH on a slide and a cover slip is placed. The slide is then gently
heated by passing through the flame 2-3 times. The slide is
observed on cooling. KOH serves to digest the protein debris and
clears keratinised tissue and increases the visibility

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 Cont.
– Calcofluor white : This is a fluorescent dye, which binds
selectively to chitin of the fungal cell wall. The specimen
then can be observed under fluorescent microscope. used
to indentify fungi in tissue specimens
– India Ink: Capsules of Cryptococcus neoformans can be
demonstrated by this negative staining technique. The
background field, rather than the organism itself, is
stained, making any capsule around the organism visible
as a halo
– Gram stain: Candida(yeast cells) is best demonstrated in
clinical specimen by Gram stain.

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CULTURE
– Media SDA (Saboraud dextrose agar). Its slightly acidic
ph5.6 which inhibits bacterial growth
– Chloramphenicol and Cycloheximide may be added to SDA
• Chloramphenicol, to reduce bacterial contamination
• Cycloheximide to reduce contamination by saprophytic fungi
– Incubated at 25-30 and 37oC to reveal dimorphism
– Yeast grows in 1-5 days
– Most cultures are considered negative for growth until after 4
wks of incubation
– Moulds identified by macro and microscopic morphology
– Yeasts identified by sugar fermentation, ability to assimilate
carbon and nitrogen sources
– Other media that can be used to isolate fungi; brain-heart
infusion agar, potato dextrose agar (PDA), Corn Meal Agar
(CMA), Malt extract agar
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• BIOCHEMICAL TESTS
• Carbohydrate fermentation studies:
– This tests whether a certain yeast ferments different
carbohydrates
• Carbohydrate assimilation studies:
– This test whether a yeast can utilize different
carbohydrates as their sole source of carbon aerobically
• API 20C System
– Freeze-dried sugars are placed into wells on a plastic
strip, yeast basal medium suspended yeast is pippeted
into wells, and incubated. Turbidity determines growth

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• SEROLOGY TESTS
• Antigen testing-Detects proteins associated with a
specific fungus. This type of test available for a
variety of fungi. Diagnose infection by specific
fungus
• Antibody testing-Detects immune response to a
specific fungus. Diagnose current or recent infection
by specific fungus; monitor treatment
– Immunodiffusion
– Counter-current immune electrophoresis
– Whole cell agglutination
– Complement fixation
– ELISA
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• MOLECULAR TECHNIQUES- species
specific
– PCR- detection of DNA in blood, serum,
broncheoalveolar lavage, sputum e.t.c
– Ribosomal DNA is hybridized to a labellled DNA
probe.

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 Special confirmatory tests
germ tube (GT) test;
• The germ tube test is used for presumptive
identification of Candida albicans. It is a rapid
screening test wherein the production of germ
tubes (pseudohyphae) within 2 hours in contact
with the serum is considered as indicative of
Candida albicans.

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 Beneficial Effects of Fungi:
• Decomposition - nutrient and carbon recycling.
• Biosynthetic factories. The fermentation property
is used for the industrial production of alcohols,
fats, citric, oxalic and gluconic acids.
• Important sources of antibiotics, such as Penicillin.
• Model organisms for biochemical and genetic
studies.
• Some fungi are edible (mushrooms).
• Yeasts provide nutritional supplements such as
vitamins and cofactors.
• Penicillium is used to flavour cheeses.
• Fungi (Leptolegnia caudate and Aphanomyces
laevis) are used to trap mosquito larvae in paddy
fields and thus help in malaria control. 28
 Harmful Effects of Fungi:

• Destruction of food, lumber, paper, and cloth.

• Animal and human and plant diseases
• Spoilage of agriculture produce such as
vegetables and cereals in the godown.

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