Objectives
Introduction of fungi
Biology of fungi
Cell structure, growth and development
Physiology of fungi
Nutrition, Temp, UV light, and water
Classification of fungi
Some terms
mycology
study of fungi
mycologists
scientists who study fungi
mycotoxicology
study of fungal toxins and their effects
mycoses
diseases caused by fungi
•100,000 species
•Only 100 human pathogens, fungi associated
diseases are rising, due to nosocomial
infections and in immunocompromised patients
(ie. HIV, diabetes, transplant folks)
•Aspergillosis, Blastomycosis—pulmonary
infections and dissemination may be involved.
•5,000 plant pathogens=$1 billion/yr
FUNGI
Fungal Characteristics
Eukaryotic
Most are Multicellular and some are unicellular.
Heterotrophic
Absorb nutrients - may be saprobes (absorb from dead
material), parasites, or mutualistic symbionts (with algae
make lichen).
Secrete powerful hydrolytic enzymes.
Cell walls contain chitin, an amino sugar polysaccharide
also found in arthropod exoskeletons
Lack flagella
Characteristics of fungi
A. eukaryotic, non- vascular organisms
B. reproduce by means of spores (conidia), usually wind-disseminated
C. both sexual (meiotic) and asexual (mitotic) spores may be
produced, depending on the species and conditions
D. typically not motile, although a few (e.g. Chytrids) have a motile phase.
E. like plants, may have a stable haploid & diploid states
F. vegetative body may be unicellular (yeasts) or multicellular moulds
composed of microscopic threads called hyphae.
G. cell walls composed of mostly of chitin and glucan.
H. Complex cytoplasm with internal organelles, microfilaments and
microtubules
H. fungi are heterotrophic ( “other feeding,” must feed on preformed
organic material), not autotrophic ( “self feeding,” make their own
food by photosynthesis).
- Unlike animals (also heterotrophic), which ingest then digest,
fungi digest then ingest.
-Fungi produce exoenzymes to accomplish this
I. Most fungi store their food as glycogen (like animals). Plants store
food as starch.
K. Fungal cell membranes have a unique sterol, ergosterol, which
replaces cholesterol found in mammalian cell membranes
L. Tubule protein—production of a different type in microtubules
formed during nuclear division.
M. Most fungi have very small nuclei, with little repetitive DNA.
N. Mitosis is generally accomplished without dissolution of the
nuclear envelope
Introduction of fungi
Eukaryotic, Heterotrophic
(chemoheterotrophic) microorganism
No chlorophyll, non-motile
Thread of cells (hyphae), transverse cell
walls (septate), hyphal anastomosis
Storage compound; glycogen
Molds
filamentous fungi
hyphae (s., hypha)
the filaments of a mold
may be coenocytic (no cross walls) or have septa
(cross walls)
mycelium (pl. mycelia)
bundles or tangled masses of hyphae
Hyphae
Hyphae are designed to increase the surface area of fungi
and thus facilitate absorption
May grow fast, up to 1 km per day, as they spread throughout
a food source
May be coenocytic, having no septa between cells, or septa
may be present with pores through which cytoplasm can flow
moving nutrients through out the fungus
Parasitic fungi have modified hyphae called
haustoria, which penetrate the host tissue but
remain outside cell membrane
The Body of a Fungus
Fungi exist mainly in the form of slender
filaments (hyphae).
long chains of cells joined end-to-end divided by
cross-walls (septa)
rarely form complete barrier
cytoplasm freely streams in hyphae
mycelium - mass of connected hyphae
grows through and penetrates substrate
MYCELIUM
Intertwined filamentous mass formed by hyphae,
visible to the unaided eye
Forms when environmental conditions are right
Vegetative mycelium: Mycelial portion remaining
INSIDE the substrate to obtain nutrition
Reproductive mycelium: Mycelial portion extends
into air ,responsible for SPORE reproduction
Introduction of fungi
Other characteristics of fungi
the ability to synthesize lysine by the -amino
adipic acid pathway (AAA-pathway)
possession of a chitinous cell wall
plasma membranes containing the sterol
ergosterol
and microtubules composed of tubulin.
Fungal wall
Shape of fungi
Protect against osmotic lysis
It the wall contains pigments (melanin)
protect the cell against ultraviolet radiation or
the lytic enzymes of other organisms
It can have antigenic properties
Cell wall components
Predominance of polysaccharides, lesser
amounts of proteins and lipids
Cell wall components
The major polysaccharides of cell wall matrix
consist of glucans such as manans, chitosan,
and galactans
Glucan refers to a group of D-glucose polymers
having glycosidic bonds
Insoluble -glucans are apparently amorphous in cell
wall
Mannans, galactomannans, rhamnomannans are
responsible for the immunologic response to the
medically important yeasts and molds
Cell wall components
Consisting of chitinous microfibrils embedded in
the matrix of small polysaccharides, proteins,
lipids, inorganic salts, and pigments
Chitin is a (1-4)-linked polymer of N-acetyl-D-
glucosamine (GlcNAc)
Produced in cytosol (from UDP GlcNAc into
chains of chitin by chitin synthetase)
The chitin microfibrils are transported to the
plasmalemma and subsequently integrated into
the new cell wall
Cell wall components
In addition to chitin, glucan, and mannan, cell
walls may contain lipid, protein, chitosan, acid
phosphatase, amylase, protease, melanin, and
inorganic ions (phosphorus, calcium, and
magnesium)
The outer cell wall of dermatophytes contains
glycopeptides that may evoke both immediate
and delayed cutaneous hypersensitivity
Plasma membrane
The main role of the plasma membrane
To regulate the uptake and release of materials
Integral membrane protein (chitin syntase,
glucan syntase)
Signal transduction
Plasma membrane
Similar to mammalian plasma membrane,
differing in having the nonpolar sterol
ergosterol, rather than cholesterol
regulates the passage of materials into and
out of the cell by being selective permeable
Several antifungal agents interfere with
ergosterol synthesis (i.e., amphotericin B)
Microtubules
Composed of the protein tubulin, which
consists of a dimer composed of two
protein subunits.
Microtubules are long, hollow cylinders ~
25 nm in diameter
Involved in the movement of organelles,
chromosomes, nuclei, and Golgi vesicle
containing cell wall precursor
Microtubules
Assist in the movement of chromosomes
during mitosis and meiosis
the destruction of cytoplasmic microtubules
interferes with the transport of secretory
materials to the cell periphery, which may
inhibit cell wall synthesis
Nucleus
The nucleus is bounded by a double nuclear
envelope and contains chromatin and a
nucleolus
Fungal nuclei are variable in size, shape, and
number
The number of chromosomes varies with the
particular fungus
S.cerevisiae ; 18 (n)
T.mentagophytes ; 4 (n)
The growth of hyphae
Apical extension
Balance between wall
synthesis and wall lysis
The apical vesicles are
produced from Golgi
bodies and then
transported to the tip
Spitzenkörper (apical body)
The vesicles fuse with the plasma membrane at the tip,
and release their contents.
enzymes involved in wall synthesis, (chitin syntase,
glucan synthase)
enzymes involved in wall lysis,
enzyme activators,
some preformed wall polymers such as mannoproteins
Aeration
The fungi include species that are obligately
aerobic (eg. most Zygomycota), obligately
anaerobic (eg. rumen fungi)
Organisms can obtain energy by oxidative
(respiratory) metabolism or by fermentation
O2 is used for oxidative metabolism to generate
energy. However it is essential for biosynthesis
of sterols, unsaturated fatty acids and some
vitamins
Table 2 Energy metabolism in relation to O2
requirements
Obligately oxidative. Obligate aerobes. Exp. Rhodotorula
Facultatively fermentative. Energy can be obtained by
oxidative and fermentative processed such fungi are
likely to be faculative anaerobes. Oxidative
metabolism, provides much more energy than
fermentative, so higher yields can occur under aerobic
conditions. Exp. Mucor, Saccharomyces
Obiligately fermentative. Oxygen is not needed for energy
production , may be either harmless or toxic. Exp.
Blastocladia, Neocallimastix
Diagrammatic
representation of the
mixed-acid fermentation
of the rumen chytrid
Neocallimastix. Part of
the fermentation occurs
in the cytosol and
hydrogenosome
Hydrogenosome:
functionally
equivalent to the
mitochondria of
aerobic organisms
Fungal nutrition
A. Absorptive mode
over whole surface or via restricted absorbing regions ,
e.g. a. rhizoids in "lower" fungi
b. substrate hyphae* in "higher" fungi
c. apical tips of hyphae
*The substrate hyphae of molds nourish the
aerial hyphae and reproductive hyphae
B. Extracellular digestion
Fungi secrete enzymes that depolymerize complex
natural products (proteins, carbohydrates, lipids, etc.) so
they can be absorbed as sources of carbon and energy.
The nutrient requirement of fungi
Carbon needs for the synthesis of
carbohydrates, lipids, nucleic acids, and
proteins.
Simple sugars, polysaccharides, citric acid, glycerol
Nitrogen for synthesis of amino acids for
proteins, purines and pyrimidines for nucleic
acids, glucosamine for chitin, and various
vitamins
Amino acid, ammonium, nitrate
Nutrition
C/N ratio (20:1)
Other elements
P : energy-rich compound metabolism,
phospholipid in lipid bilayer
K : coenzyme
Mg : concer with sporulation
S : protein component
Trace elements
Fe, Cu, Mn, and Zn
Czapek-Dox medium
widely used for the culture of fungi
Mineral base:
C and energy source:
N source:
Water:
If a solid medium is
required:
KH2PO4
MgSO4.7H2O
KCl
FeSO4.7H2O
Sucrose (Glu,starch)
NaNO3
Agar
1 g
0.5 g
0.5 g
0.01 g
30 g
2 g
1 litre
20 g
Water availability
Most fungi require very high water availability
(relative humidity), and rapidly dry out or
senescence in dry conditions.
Water activity (aw) = ps/pw (pure water = 1)
DNA is denatured at aw = 0.55
Osmophiles 0.85, Xerophiles 0.80, Halophiles 0.75
The xerotolerant fungi can grow slowly, at water
activity of 0.64.
Hydrogen ion
Opt. pH 5.0-7.0
Acid-tolerant (pH 2.0) Aspergillus,
Penicillium, Fusarium, yeast in stomach of
animals
Strongly alkaline environment (pH 10-11)
F.oxysporum, P.variabile
Light
Influence on fungal growth in specific cases
light does not play a major part in growth
and metabolism of fungi
A common metabolic effect of light is the
induction of carotenoid biosynthesis
Morphology
Yeast
Unicellular, round or oval, size 8-15 x 3-5 µm
Conidiogenesis (budding, binary fission, sexual
spores)
Budding
yeasts
Binary fission
Morphology
Dimorphic fungi (thermally dimorphic fungi)
Environment/Routine culture media
(SDA) 25-300C ---Mold form
Tissue/Enriched media (BHI)
35-370C---Yeast form
Sporothrix schenckii
CLASSIFICATION
Fungi are eukaryotic organisms that do
not contain chlorophyll, but have cell walls,
filamentous structures, and produce spores.
These organisms grow as saprophytes and
decompose dead organic matter. There are
between 100,000 to 200,000 species depending on
how they are classified. About 300species are
presently known to be pathogenic for man. There
are five kingdoms of living things. The fungi are
in the Kingdom Fungi.
Medically important fungi are in four phyla:
1. Ascomycota - Sexual reproduction in a sack called an ascus
with the production of ascopspores.( Aspergillus, Blastomyces
dermatidis, Histoplasma capsulatus)
2. Basidiomycota -Sexual reproduction in a sack called a
basidium with the production of basidiospores.( Cryptococcus
neoformans)
3. Zygomycota - sexual reproduction by gametes and asexual
reproduction with the formation of zygospores.( Rhizopus,
Mucor)
4. Deuteromycota(Fungi Imperfecti Mitosporic Fungi) - no
recognizable form of sexual reproduction. Includes most
pathogenic fungi ( Sporothrix, Coccidioides immitis, Candida,
Pneumocystis).