PHA6114

PHARMACEUTICAL MICROBIOLOGY AND

PARASITOLOGY

FUNGI
ASST. PROF. RHONA P. RAMOS, MSc.
Course Facilitator
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eukaryotic organisms
many similarities between the
biochemistry of fungal cells and
vertebrate (human) cells

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part of the normal flora of warm-blooded animals
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decomposers of organic matter

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animal and plant pathogens

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mycosis /mīˈkōsəs/
a disease caused by infection with a fungus

SUPERFICIAL
CUTANEOUS
SUBCUTANEOUS
SYSTEMIC
OPPORTUNISTIC

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SUPERFICIAL

Pityriasis versicolor White piedra

Malassezia furfur Trichosporon beigelii
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 CUTANEOUS

Tinea pedis (athlete’s foot) Onychomycosis (nail infection) Tinea capitis (scalp ringworm)
Trichophyton rubrum Trichophyton rubrum Trichophyton tonsurans

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SUBCUTANEOUS

Chromoblastomycosis Mycetoma
Fonsecaea pedrosoi Acremonium spp.

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SYSTEMIC

Blastomycosis Histoplasmosis
Blastomyces dermatitidis Histoplasma capsulatum

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SYSTEMIC

Coccidioidomycosis Paracoccidioidomycosis
Coccidioides immitis Paracoccidioides brasiliensis

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OPPOTUNISTIC Candidosis (superficial/systemic)
Candida albicans
Candida glabrata
Candida parapsilosis

Pneumonia
Aspergillosis
Pneumocystis jirovecii
Aspergillus fumigatus
(carinii)
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economically important fungi
great benefit to humanity

FILAMENTOUS FUNGI
YEAST

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FILAMENTOUS FUNGI

edible mushroom Sake (rice wine)

Agaricus bisporus Aspergillus sp. + Saccharomyces sp

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FILAMENTOUS FUNGI

Single cell protein Penicillin production

Fusarium graminearum Penicillium chrysogenum

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FILAMENTOUS FUNGI

Cheese flavouring
(Roqueforti ‘blue’ cheese)
Penicillium roqueforti

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YEAST

Bakers’ yeast—bread
Brewers’ yeast—beer, wine, cider, etc.
Enzyme (invertase)
Gene expression system
Dietary supplement
Saccharomyces cerevisiae

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 Oomycetes Ascomycetes Basidiomycetes
mildews and water yeast species mushrooms and
moulds Saccharomyces cerevisiae bracket fungi

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 Teliomycetes Ustomycetes Deuteromycetes
Aspergillus,
rust fungi (plant smuts (plant
Fusarium and
pathogens) pathogens)
Penicillium

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 FUNGAL CELL

oval in shape
rigid cell wall - glucan,
mannan, chitin
protoplast

AVAILABLE VIDEO CLIP INSIDE LEARNING MODULE

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FUNGAL CELL

Periplasmic space
contains secreted
proteins that do not
penetrate the cell wall

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FUNGAL CELL
cell membrane or
plasmalemma
phopholipid bilayer which
contains phospholipids, lipids,
protein and sterols
10 nm
ergosterol - target of
amphotericin B

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FUNGAL CELL

nucleus
surrounded with nuclear
membrane with pores
contain histones

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FUNGAL CELL

chromosomes
vary in size from 0.2 to 6Mb
number varies per type of yeast

S. cerevisiae

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FUNGAL CELL

plasmids
also in the nucleus
extrachromosomal information
ex. 2 µm plasmid, killer plasmids

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FUNGAL CELL

mitochondrion
powerhouse of the cell
enzymes of the
tricarboxylic acid cycle
(Krebs’ cycle) are located
in the matrix

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FUNGAL CELL

mitochondrion
inner membrane - electron
transport and oxidative
phosphorylation
outer membrane - contains
enzymes involved in lipid
biosynthesis

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FUNGAL CELL

mitochondrion
semi- independent
organelle as it possesses its
own DNA
capable of producing its own
proteins on its own
ribosomes which are referred
to as mitoribosomes

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FUNGAL CELL

ribosomes
site of protein biosynthesis
in the form of polysomes—
lines of ribosomes strung
together by a strand of
mRNA

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FUNGAL CELL
Golgi apparatus
endoplasmic reticulum
plasmalemma

export of proteins

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FUNGAL CELL
vacuole
‘storage space’ for nutrients,
hydrolytic enzymes or
metabolic intermediates

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MEDICAL SIGNIFICANCE
FUNGAL PATHOGENS
yeasts
moulds
dermatophytes

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MEDICAL SIGNIFICANCE
FUNGAL PATHOGENS

yeasts
C. albicans

most frequently encountered human fungal pathogen

superficial and systemic infections

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MEDICAL SIGNIFICANCE
FUNGAL PATHOGENS

yeasts
C. albicans

superficial
oropharyngeal and genital conditions
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MEDICAL SIGNIFICANCE
FUNGAL PATHOGENS

yeasts
C. albicans

superficial
oropharyngeal and genital conditions
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MEDICAL SIGNIFICANCE
FUNGAL PATHOGENS

mould
Aspergillus fumigatus
dominant fungal pulmonary pathogen of humans
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MEDICAL SIGNIFICANCE
FUNGAL PATHOGENS

mould
Aspergillus fumigatus

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MEDICAL SIGNIFICANCE
FUNGAL PATHOGENS
dermatophyte - range of fungi capable of colonizing the skin,
nails or hair
Trichophyton, Microsporum, Epidermophyton

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ANTIFUNGAL THERAPY
POLYENE ANTIFUNGALS
large macrolide ring

macrolide \ˈmak-rə-ˌlīd\
any of several antibiotics (as erythromycin or clarithromycin)
that contain a usually 14- to 16-membered macrocyclic lactone
ring

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ANTIFUNGAL THERAPY
POLYENE ANTIFUNGALS

large number of hydroxyl groups distributed along the

macrolide ring on alternate carbon atoms

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ANTIFUNGAL THERAPY
POLYENE ANTIFUNGALS

amphotericin B

produced by the bacterium Streptomyces nodosus

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ANTIFUNGAL THERAPY
POLYENE ANTIFUNGALS

amphotericin B

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ANTIFUNGAL THERAPY
POLYENE ANTIFUNGALS

amphotericin B

SE: renal damage during prolonged antifungal therapy

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ANTIFUNGAL THERAPY
POLYENE ANTIFUNGALS

nystatin

same mode of action as amphotericin B

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS
still the most widely used group of antifungal agents

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 ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

azole derivatives
are classified as
imidazoles or
triazole

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

interferes with ergosterol biosynthesis by binding to the cytochrome

P-450 mediated enzyme known as 14-α-demethylase (P-450DM)
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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS
build-up of toxic intermediates which can prove fatal to
the fungal cell

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS
miconazole
first azole used to treat systemic
fungal infections but
demonstrated a number of toxic
side effects

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

ketoconazole
produced high serum concentrations upon oral
administration but had poor activity against aspergillosis

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS
fluconazole
highly effective in the treatment of
infections caused by C. albicans
but shows limited activity against
Aspergillus

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

itraconazole
proven efficacy against Aspergillus
exhibits both fungicidal and
fungistatic effects

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

itraconazole
extensive hepatic metabolism which yields up to 30
metabolites, a number of which retain antifungal activity
IV - for severe Aspergillus infection
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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS
voriconazole, posaconazole and ravuconazole

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

voriconazole
good activity against pulmonary aspergillosis and
cerebral aspergillosis

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ANTIFUNGAL THERAPY
AZOLE ANTIFUNGALS

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ANTIFUNGAL THERAPY
ECHINOCANDINS

semisynthetic lipopeptides comprising a cyclic

hexapeptide core connected to a lateral fatty acid chain
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ANTIFUNGAL THERAPY
ECHINOCANDINS

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ANTIFUNGAL THERAPY
ECHINOCANDINS

target the synthesis of β-1,3-glucan, the major polymer

of the fungal cell wall
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ANTIFUNGAL THERAPY
ECHINOCANDINS

caspofungin
fungistatic against different Aspergillus species
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ANTIFUNGAL THERAPY
ECHINOCANDINS

caspofungin
fungicidal against a range of Candida species
including species that are resistant (e.g. C. krusei) or
isolates that are less susceptible (e.g. C. dubliniensis,
C. glabrata) to azoles, or resistant to amphotericin B

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ANTIFUNGAL THERAPY
ECHINOCANDINS

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ANTIFUNGAL THERAPY
ECHINOCANDINS

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ANTIFUNGAL THERAPY
ECHINOCANDINS

AVAILABLE VIDEO CLIP

ABOUT MOA OF
ANTIFUNGAL DRUGS
INSIDE LEARNING MODULE

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ANTIFUNGAL THERAPY
SYNTHETIC ANTIFUNGAL AGENTS

flucytosine
synthetic fluorinated pyrimidine
good activity against a range of yeast
species and moderate levels of activity
against Aspergillus species

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ANTIFUNGAL THERAPY
SYNTHETIC ANTIFUNGAL AGENTS
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ANTIFUNGAL THERAPY
SYNTHETIC ANTIFUNGAL AGENTS

flucytosine
yeast cells increase in size
when exposed to levels lower
than the minimum inhibitory
concentration (MIC)

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ANTIFUNGAL THERAPY
SYNTHETIC ANTIFUNGAL AGENTS

flucytosine
resistance has been attributed to alteration in the
enzyme (cytosine deaminase) or to an elevation in
the amount of pyrimidine synthesis

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MEDICALLY IMPORTANT FUNGAL PATHOGENS
Candida albicans

opportunistic fungal pathogen

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 Candida albicans

an antifungal drug-resistant biofilm

structure secreted as small, spherical
extracellular vesicles (arrows) that
deliver payloads of biofilm polymers
to the extracellular matrix

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GIT infection in diabetics, cancer patients
and people with AIDS
esophagus is a common site of infection,
rendering swallowing difficult

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Candida albicans

urinary tract can be a site of candidosis which may be due

to renal infection, other underlying disease(s) or cystitis
indwelling urinary catheter may also predispose to
Candida infection
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Candida albicans

Factors which impair the host’s immune system:

underlying disease (AIDS, cancer, diabetes)
use of immunosuppressive therapy during organ
transplantation
broad-spectrum antibiotic therapy

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Candida albicans

virulence factors
are essential for the invading organism to colonize,
invade host tissue, adapt to various environments in
the host, subvert host functions, and overcome the
defenses of the host

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Candida albicans
virulence factors
1. ability to adhere

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Candida albicans

virulence factors

2. yeast to hyphal morphogenesis

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Candida albicans

hyphae
capable of thigmotropism
(contact sensing) which may
aid in finding the line of least
resistance between and
through layers of cells in
tissue

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Candida albicans

virulence factors
3. production of extracellular enzymes
Phospholipases A, B, C and
lysophospholipase - damage host cell
membranes and facilitate invasion

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Candida albicans

virulence factors
3. production of extracellular enzymes
acid proteinases - degradation of the
immunoglobulins IgG and IgA
secreted aspartic proteinase (SAP)

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Candida albicans

virulence factors
3. production of extracellular enzymes
haemolysin - to access iron released from
ruptured red blood cells

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Candida albicans

virulence factors
4. ability to bind to platelets via fibrinogen binding ligands

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Candida albicans

virulence factors
5. phenotypic switching
allows the yeast to exploit its ‘dominant’ or
‘controlling’ virulence factor with antifungal
drug resistance, adherence, extracellular
enzyme production

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tissue colonization and
invasion

adherence - enzymes (phospholipase and

proteinase) can facilitate adherence by
damaging or degrading cell membranes
and extracellular proteins
tissue colonization and
invasion

thigmotropism - produce hyphae to find

the line of least resistance

degradation of tissue
tissue colonization and
invasion

enter the host’s blood stream

burst blood cells and release iron -

haemolysin
tissue colonization and
invasion

escape from the bloodstream - adherence

to the walls of capillaries and passage
across the wall
MEDICALLY IMPORTANT FUNGAL PATHOGENS
Aspergillus fumigatus

saprophytic fungus
widely distributed in nature
opportunistic pathogen

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Aspergillus fumigatus

pulmonary aspergillosis
(80–90% of cases)
greater mortality rates
compared to diseases
due to Candida

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Aspergillus fumigatus
virulence factors
1. produce extracellular enzymes
Phospholipase - for tissue degradation and may
facilitate exit of the fungus from the lung into the
bloodstream

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Aspergillus fumigatus
virulence factors
1. produce extracellular enzymes
Proteases

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Aspergillus fumigatus
virulence factors
1. produce extracellular enzymes
Proteases
Elastinase
a. serine protease and a
metalloproteinase
Elastin constitutes SP: also function as allergens which
almost 30% of lung may be important in the induction and
tissue persistence of allergic aspergillosis

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Aspergillus fumigatus
virulence factors
1. produce extracellular enzymes
Proteases
may cause local inflammation results in
airway damage
capable of inducing epithelial cell
detachment (desquamation) from basement
membranes

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Aspergillus fumigatus
virulence factors
1. produce extracellular enzymes
Proteases
induce the release of the proinflammatory
IL-6 and IL-8 cytokines, which may induce
mucosal inflammatory response and
subsequent damage to the surrounding
tissue

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Aspergillus fumigatus
virulence factors
2. production and secretion of toxins
gliotoxin - main toxin
helvolic acid, fumigatin and fumagillin
act to retard elements of the local immunity

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MEDICALLY IMPORTANT FUNGAL PATHOGENS
Histoplasma capsulatum

histoplasmosis - most prevalent fungal pulmonary infection

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Histoplasma capsulatum
dimorphic fungus
1. mycelial in soil
2. round, budding cells in tissue

natural habitat is soil that has been enriched with the droppings of bats or birds
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Histoplasma capsulatum

individuals develop a non-

specific flu-like illness associated
with fever, chills, headache and
chest pains after a 3 week
incubation period that resolves
without treatment

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MEDICALLY IMPORTANT FUNGAL PATHOGENS
Crypotococcus neoformans

encapsulated yeast
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Crypotococcus neoformans

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Crypotococcus neoformans

facultative intracellular
pathogen

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 Crypotococcus neoformans
virulence factors

1. produces melanin -
ability to bind and
protect against
microbicidal peptides

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Crypotococcus neoformans
virulence factors
2. capsule
it protects the cell from the
immune response
capsular material
(glucuronoxylomannans) induces
the shedding of host cell
adherence molecules

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Crypotococcus neoformans

cryptococcosis may be controlled by antifungal

therapy, in AIDS patients there is a danger of relapse
unless antifungal therapy is constantly maintained

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MEDICALLY IMPORTANT FUNGAL PATHOGENS
DERMATOPHYTES

a group of keratinophilic fungi

which can metabolize keratin
—the principal protein in skin,
nails and hair

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DERMATOPHYTES
tinea capitis
infection of the hair and scalp with
a dermatophyte, usually
Microsporum canis or Trichophyton
violaceum

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DERMATOPHYTES
tinea corporis

usually caused by Trichophyton spp., Microsporum sp.

or Epidermophyton floccosum

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DERMATOPHYTES
tinea cruris

usually Epidermophyton
floccosum or Trichophyton
rubrum

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DERMATOPHYTES

tinea pedis
T. rubrum and E.
floccosum

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DERMATOPHYTES
tinea manuum
Trichophyton mentagrophytes

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DERMATOPHYTES
tinea unguium
infection of the fingernails or toenails and is
often described as onychomycosis

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EMERGING FUNGAL PATHOGENS
Saccharomyces cerevisiae
most studied and best-characterized organism on the planet

brewer’s or baker’s yeast

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Saccharomyces cerevisiae

Biosafety Level 1 pathogen, indicating that it can cause

superficial or mild systemic infections in certain instances

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Saccharomyces cerevisiae

vulvovaginitis
pneumonia
septicaemia
postoperative peritonitis
fever and coughs in transplant patients
lung infection

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Saccharomyces cerevisiae

virulence factors
1. ability to grow at 42 °C
febrile patients can attain this temperature and it
is advantageous for a pathogenic microorganism
to survive at this elevated temperature

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Saccharomyces cerevisiae

virulence factors
2. extracellular enzymes
acid proteinases - modulating the immune system’s
response
phospholipases - degradation of cell membranes

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Saccharomyces cerevisiae

virulence factors
3. ability to grow in a pseudohyphal form which may assist
in the penetration of tissue
4. obstructing capillaries particularly in the brain with
concomitant damage to surrounding tissue - flocculation

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Saccharomyces cerevisiae

virulence factors
5. adhere to epithelial tissue via a proteinaceous adhesin
6. ability to alter their phenotype
contributes to the yeast’s persistence in the body

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Saccharomyces cerevisiae

virulence factors
conventional therapy - azoles or polyenes
resistance to fluconazole
mediated via a multidrug efflux pump

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EMERGING FUNGAL PATHOGENS
Non-albicans Candida species
Candida dubliniensis

C. albicans

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EMERGING FUNGAL PATHOGENS
Non-albicans Candida species
Candida krusei
demonstrates a reduced ability to adhere to
epithelial cells compared to C. albicans
display a high cell surface hydrophobicity which
allows it stick to and colonize catheters and
implants
resistance to fluconazole

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Candida krusei

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EMERGING FUNGAL PATHOGENS
Non-albicans Candida species
Candida glabrata
fourth most commonly isolated Candida species
partially resistant to fluconazole

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EMERGING FUNGAL PATHOGENS
Penicillium marneffei

asexual, dimorphic fungus growing as a mycelium at

37 °C in tissue and as single cells at 28 °C
PCR (polymerase chain reaction) fingerprinting is used
to identify

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Penicillium marneffei

pulmonary involvement is often seen particularly in

AIDS patients
fever, weight loss, anaemia, skin lesions, and liver and
spleen inflammation
fatal if untreated with antifungal drugs

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ANTIBIOTIC PRODUCTION
penicillin, Penicillium notatum
first member of the β-lactam class of antibiotics

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 ANTIBIOTIC PRODUCTION
penicillin, Penicillium notatum
for killing Gram-positive bacteria
inhibits peptidoglycan synthesis

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ANTIBIOTIC PRODUCTION

penicillin, Penicillium notatum

Penicillium chrysogenum -
as superior producer
three types, namely F, G
and V

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ANTIBIOTIC PRODUCTION

majority are produced by fermentation and most are

secondary metabolites
occurs in the stationary phase and is linked to
sporulation
use low levels of glucose in the fermentation medium or
to obtain a mutant which is not catabolite repressed

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ANTIBIOTIC PRODUCTION
chemical content of the medium must be monitored
since high levels of nitrogen or phosphate (PO4) retard
antibiotic production
feedback inhibition, where the antibiotic builds to high
intracellular levels and retards production or kills the cell
introduce low levels of the antifungal agent
amphotericin B

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