SHOP 5

PAT 1

MCB 305(MYCOLOGY) LECTURE NOTE

INTRODUCTION TO MYCOLOGY

The term "mycology" is derived from Greek word "mykes" meaning mushroom. Therefore
mycology is the study of fungi.

The ability of fungi to invade plant and animal tissue was observed in early 19th century but the
first documented animal infection by any fungus was made by Bassi, who in 1835 studied the
muscardine disease of silkworm and proved the that the infection was caused by a fungus
Beauveria bassiana. In 1910 Raymond Sabouraud published his book Les Teignes, which was a
comprehensive study of dermatophytic fungi. He is also regarded as father of medical mycology.

Fungi were initially classified with plants and were a subject of interest for botanists; hence the
influence of botany can be seen on their classification. In 1969 R.H Whittaker classified all
living organisms into five kingdoms namely Monera, Protista, Fungi, Plantae and Animalia.
Traditionally the classification proceeds in this fashion: Kingdom - Subkingdom - Phyla/phylum
- Subphyla - Class - Order - Family - Genus- Species

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 and are therefore non-motile, a feature that separates them from animals.
All fungi possess cell wall made of chitin. 4. Are chemoheterotrophs (require organic
compounds for both carbon and energy sources) and fungi lack chlorophyll and are therefore not
autotrophic.

5. Fungi are osmiotrophic; they obtain their nutrients by absorption.

6. They obtain nutrients as saprophytes (live off of decaying matter) or as parasites (live off of
living matter).

7. All fungi require water and oxygen and there are no obligate anaerobes.

8. Typically reproduce asexually and/or sexually by producing spores.

9. They grow either reproductively by budding or non-reproductively by hyphal tip elongation.

10. Food storage is generally in the form of lipids and glycogen.

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Classification of fungi:

Based on sexual reproduction(systemic) and the other based on morphology of the thallus
(vegetative structure).

Fungi exist in two fundamental forms; the filamentous (hyphal) and single celled budding forms
(yeast). But, for the classification sake they are studied as moulds, yeasts, yeast like and
dimorphic fungi.

1. Moulds:

The thallus of mould is made of hyphae, which are cylindrical tube like structures that elongates
by growth at tips. A mass of hyphae is known as mycelium. It is the hypha that is responsible for
the filamentous nature of mould. The hyphae may be branched or unbranched. They may be
septate or aseptate. Hyphae usually have cross walls that divide them into numerous cells. These
cross walls, called septa have small pores through which cytoplasm is continuous throughout the
hyphae. Therefore all hyphal fungi tend to be coenocytic (multinucleate). With exception of
zygomycetes (Rhizopus, Mucor), all moulds are septate. Non-septate hyphae are considered to
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be more primitive because if a hyphal strand is damaged the entire strand dies. When a septate
hyphal strand is damaged, the pores between adjacent compartments can be plugged, thus
preventing death of the whole hyphal strand.

Mycelium are of three kinds: 1. Vegetative mycelium are those that penetrates the surface of the
medium and absorbs nutrients.

2. Aerial mycelium are those that grow above the agar surface

3. Fertile mycelium are aerial hyphae that bear reproductive structures such as conidia or
sporangia.

Since hypha is the structural unit of mould, the mycelium imparts colour, texture and topography
to the colony. Those fungi that possess melanin pigments in their cell wall are called phaeoid or
dematiaceous and their colonies are coloured grey, black or olive. Examples are species of
Bipolaris, Cladosporium, Exophiala, Fonsecaea, Phialophora and Wangiella Those hyphae that
don't possess any pigment in their cell wall are called hyaline.

Hyphae may have some specialised structure or appearance that aid in identification. Some of
these are:

a) Spiral hyphae: These are spirally coiled hyphae commonly seen in Trichophyton
mentagrophytes.

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b) Pectinate body: These are short, unilateral projections from the hyphae that resemble a broken
comb. Commonly seen in Microsporum audouinii.

c) Favic chandelier: These are the group of hyphal tips that collectively resemble a chandelier or
the antlers of the deer (antler hyphae). They occur in Trichophyton schoenleinii and
Trichophyton violaceum.

d) Nodular organ: This is an enlargement in the mycelium that consists of closely twisted
hyphae. Often seen in Trichophyton mentagrophytes and Microsporum canis.

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e) Racquet hyphae: There is regular enlargement of one end of each segment with the opposing
end remaining thin. Seen in Epidermophyton floccosum, Trichophyton mentagrophytes.

f) Rhizoides: These are the root like structures seen in portions of vegetative hyphae in some
members of zygomycetes.

2. Yeasts:

Yeasts are unicellular spherical to ellipsoid cells. They reproduce by budding, which result in
blastospore (blastoconidia) formation. In some cases, as the cells buds the buds fail to detach and
elongate thus forming a chain of elongated hyphae like filament called pseudo hyphae. This
property is seen in Candia albicans. The same species also have the ability to produce true
hypha, which is seen as germ tube. The difference between the two is that there is a constriction
in pseudo hyphae at the point of budding, while the germ tube has no constriction.

Some yeast such as Cryptococcus and the yeast form of Blastomyces dermatatidis produce
polysaccharide capsule.

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Capsules can be demonstrated by negative staining methods using India ink or Nigrosine. The
capsule itself can be stained by Meyer Mucicarmine stain. Some yeasts are pigmented.
Rhodotorula spp produces pink colonies due to carotenoid pigments while some yeasts such as
Phaeoannellomyces werneckii and Piedraia hortae are dematiaceous, producing brown to
olivaceous colonies. True yeasts such as Saccharomyces cerviciae don't produce pseudo hyphae.

3. Yeast like:
These are fungi which occur in the form of budding yeast-like cell. Similar to yeasts but
produce pseudohyphae. Yeast-like fungi may be basidiomycetes, such as Cryptococcus
neoformans or ascomycetes such as Candida albicans.

4. Dimorphic:
Fungi existing in two different morphological forms at two different environmental
conditions. They exist as yeasts in tissue and in vitro at 37oC and as moulds in their
natural habitat and in vitro at room temperature. Eg: Histoplasma capsulatum,
Blastomyces dermatidis, Paracoccidiodes brasiliensis, Coccidioides immitis

Reproduction in fungi:

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Fungi reproduce by:

Asexual and sexual methods.

Asexual reproduction: this is the commonest mode in most fungi with fungi participating in
sexual mode only under certain circumstances.

The form of fungus undergoing asexual reproduction is known as anamorph (or imperfect stage)
and when the same fungus is undergoing sexual reproduction, the form is said to be teleomorph
(or perfect stage). The whole fungus, including both the forms is referred as holomorph.
(Taxonomically, the teleomorph or the holomorph is used, but practically it is more convenient
to use the anamorph.)

Asexual propagules are termed either spores or conidia depending on their mode of production.
Asexual spores are produced following mitosis whereas sexual spores are produced following
meiosis.

The asexual spores of zygomycetes, which are known as sporangiospores form within sac like
structure known as sporangia. The sporangiospores result from the mitotic cleavage of cytoplasm
in the sporangium. The sporangia are borne on special hyphae called sporangiophore. This
endogenous process of spore formation within a sac is known as sporogenesis.

Conidia arise either by budding off conidiogenous hyphae or by differentiation of preformed

hyphae. These develop following mitosis of a parent nucleus and are formed in any manner
except involving cytoplasmic cleavage. This exogenous process is known as conidiogenesis, a
process that occurs both in yeasts and moulds. Conidia are borne on specialised structures called
conidiophore.

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• Microconidia - Small, single celled

• Macroconidia – Large and septate and are often multicellular

Conidia production may be blastic or thallic.

In blastic development the conidium begins to enlarge and a septum is formed. Here the
conidium originates from part of parent.

In thallic mode of development the conidium is differentiated by a septum before its

differentiation. Thus the conidium results from the conversion of entire parent cell into the
conidium.

The cell that gives rise to a conidium is called a conidiogenous cell. Conidiophores are
specialised hyphae that bear conidia or conidiogenous cells. In many cases conidiogenous cells
are referred as phialides.

• Vegetative spores:

• Blastospores: These are formed by budding from parent cell, as in yeasts

• Arthrospores – formed by segmentation & condensation of hyphae

• Chlamydospores –thick walled resting sporese.g. C.albicans

Sexual Reproduction: Sexual propagules are produced by the fusion of two nuclei that then
generally undergo meiosis.

The first step in sexual methods of reproduction involves plasmogamy (cytoplasmic fusion of
two cells).

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The second step is karyogamy (fusion of two compatible nuclei), resulting in production of
diploid or zygote nucleus.

This is followed by genetic recombination and meiosis. The resulting four haploid spores are
said to be sexual spores, e.g. zygospores, ascospores and basidiospores.

If a sexual spore is produced only by fusion of a nucleus of one mating type with a nucleus of
another mating type (+ and - strains), the fungus is said to be heterothallic.

In contrast, homothallic moulds produce sexual spores following the fusion of two nuclei from
the same strain (+ and + or - and - strains).

Based on sexual spores formation: 4 classes

1. Zygomycetes: Commonly known as bread moulds, these are fast growing, terrestrial,
largely saprophytic fungi. Hyphae are coenocytic and mostly aseptate, lower fungi.

Asexual spores -Sporangiospores: presentwithin a swollen sac- like

structure called sporangium .Examples: Rhizopus, Absidia, Mucor

2. Ascomycetes: Sexual spores called ascospores are present within a sac like structure
called Ascus. Each ascus has 4 to 8 ascospores. Includes both yeasts and filamentous
fungi.
Narrow, septate hyphae. Asexual spores are called conidia borne on conidiophore.
Examples: Penicillium, Aspergillus

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3. Basidiomycetes: They exist as saprobes and parasites of plants. Hyphae are dikaryotic
and can often be distinguished by the presence of clamp connections over the septa.
Sexual fusion results in the formation of a club shaped organ called base or basidium
which bearspores called basidiospores
Examples: Cryptococcus neoformans, mushrooms

4. Deuteromycetes or Fungi imperfectii:

Group of fungi whose sexual phases are not identified. Grow as molds as well as yeasts.
Most fungi of medical importance belong to this class. Examples: Coccidioides immitis,
Paracoccidioides brasiliensis, Candida albicans. Deuteromycetes are also known as
Fungi Imperfecti because of absence of sexually reproducing forms (teleomorph or perfect
stage). As their teleomorph continue to be discovered, they would be classified among the
previous categories, until then this remains an artificial and heterogeneous group.

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There are three classes of Fungi Imperfecti.

1. Blastomycetes: These include asexual budding forms of Cryptococcus, Candida,

Torulopsis and Rhodotorula. Depending on the presence of melanin in their cell walls,
they may be non-dematiaceous or dematiaceous.

2. Hyphomycetes: A class of mycelial moulds which reproduce asexually by conidia on

hyphae. Hyphae are septate. This class contains the majority of medically important fungi.
Dematiaceous hyphomycetes are those conidial fungi that produce dark brown, green-
black, or black colonies. Hyaline hyphomycetes include those conidial fungi, which are
not darkly pigmented; colonies may be colourless or brightly coloured.

3. Coelomycetes: These produce acervuli, which are tightly bound mats of hyphae on
which conidia are produced.

Importance of Spores:

1) Allows for dissemination

2) Allows for reproduction

3) Allows the fungus to move to new food source.

4) Allows fungus to survive periods of adversity.

5) Means of introducing new genetic combinations into a population

6) Rapid identification (also helps with classification)

7) Source of inocula for human infection

8) Source of inocula for contamination

ECONOMIC IMPORTANCE OF FUNGI

Fungi are one of the most significant organisms in the environment. They are hidden from the
naked eyes but their effects and impact are very evident and plenty. They are the primary
decomposers of substances in the ecological system.

Beneficial Effects of Fungi:

1. Decomposition - nutrient and carbon recycling.

2. Biosynthetic factories. The fermentation property is used for the industrial production of
alcohols, fats, citric, oxalic and gluconic acids.

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3. Important sources of antibiotics, such as Penicillin.

4. Model organisms for biochemical and genetic studies. Eg: Neurospora crassa

5. Saccharomyces cerviciae is extensively used in recombinant DNA technology, which includes

the Hepatitis B Vaccine.

6. Some fungi are edible (mushrooms).

7. Yeasts provide nutritional supplements such as vitamins and cofactors.

8. Penicillium is used to flavour Roquefort and Camembert cheeses.

9. Ergot produced by Claviceps purpurea contains medically important alkaloids that help in
inducing uterine contractions, controlling bleeding and treating migraine.

10. Fungi (Leptolegnia caudate and Aphanomyces laevis) are used to trap mosquito larvae in
paddy fields and thus help in malaria control.

Harmful Effects of Fungi:

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

2. Animal and human diseases, including allergies.

3. Toxins produced by poisonous mushrooms and within food (Mycetism and Mycotoxicosis).

4. Plant diseases.

5. Spoilage of agriculture produce such as vegetables and cereals in the godown.

6. Damage the products such as magnetic tapes and disks, glass lenses, marble statues, bones
and wax.

ASSINGMENT

1. What do you understand by the term diplanetism?

2. Briefly describe the life cycle of Saprolegnia ferax to illustrate the phenomenon of
diplanetism
3. Compare the life cycle of Aspergillus sp with that of Saccharomyces cerevisae

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PAT 2
MCB 305(MYCOLOGY) LECTURE NOTE
ECONOMIC IMPORTANCE OF FUNGI
Fungi are one of the most significant organisms in the environment. They are
hidden from the naked eyes but their effects and impact are very evident and
plenty. Fungi are a group of eukaryotic organisms. They include microorganisms
like moulds, yeasts, and mushrooms. Fungi are very different from other living
organisms. One thing that separates them from other living cells is the presence of
chitin in their cell walls. Like other heterotrophs, they get their food by secreting
digestive enzymes into their environment to dissolve molecules and then absorb
them. They are the primary decomposers of substances in the ecological system.

There are different types of fungi within the fungi kingdom. Based on their
structure, there are three types of fungi commonly seen.

Yeasts: They are unicellular fungi. Examples in this category include:

● Saccharomyces cerevisiae, also known as Brewer’s or baker’s yeast.

● Saccharomyces boulardii – They can be found in the intestine as normal flora.

● Pityrosporum ovale – Can be found on the skin as normal flora. Has the ability to
become pathogenic.

● Candida Albicans – It is also present in the mouth, oesophagus, bowel, and axil.
Can also become infectious if the immunity system is compromised.

● Cryptococcus neoformans

Moulds: They are a multicellular group of fungi. Common in this group are:

● Penicillium

● Aspergillus

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● Mucor

● Rhizopus

Moulds can grow on walls, paper, leather, clothes, plants, etc. When this happens,
it is called mildew.

Mushrooms: These are a fleshy fungus that grows on soil and trees.

FUNGAL METABOLITES

Metabolism is the conversion in the body of one chemical compound into another
compound. The resulting compound is called a metabolite. A metabolite is any
substance that is produced during metabolism or that takes part in metabolism.

Primary metabolites are directly involved in normal growth, development and

reproduction. It usually performs physiological functions in the organism.
Examples include carbohydrates, proteins, amino acids, vitamins, acetone, lactic
acid, ethanol etc.

Secondary metabolites are defined as heterogenous low-molecular- weight organic

compounds that are not directly involved in primary metabolic processes such as
cell growth, cell division, cell respiration or photosynthesis and they are formed
during the end or near the stationary phase of growth . Furthermore, secondary
metabolites are derived from a few common biosynthetic pathways which branch
off the primary metabolic pathways and are often produced as families of related
compounds, often specific for a group of organisms. Many organisms produce
large numbers of secondary metabolites and the complexity and diversity is
sometimes astonishing. Secondary metabolites have had great impact on society
for centuries in traditional medicine and in modern times, as pharmaceuticals,
fragrances in cosmetics, flavouring in foods and drinks, agrochemicals, etc.

But what is the biological reason for producing these compounds? Why do
organisms put huge amounts of energy on producing them? Secondary metabolites
perform many different functions for their producer including functions as

(1) volatile alarm pheromones,

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(2) sex attractants in many insects,

(3) defense against predators,

(4) competition and toxicity against other microbes and

(5) pathogenicity.

However, many of the biological functions are still to be determined. Examples of

secondary metabolites are ergot alkaloids, antibiotics, naphthalene’s, nucleosides,
quinolones, terpenoids peptides etc

Fungi are a rich source of secondary metabolites and have been of interest for
humans for thousands of years. However, it was not until when Alexander Fleming
discovered the antibiotic penicillin (penicillin F) 1929 and its development into a
medicine by Florey and Chain in the 1940s (penicillin G), that the attention to
fungal secondary metabolites became widespread. Since then, huge research efforts
have been made and resulted in thousands of characterized compounds with a
range of different biological activities.

FUNGI IN INDUSTRIES

Fermentation and Yeast: Yeast is a single-celled fungus

Yeast can respire: with oxygen (called aerobic respiration) and without oxygen
(called anaerobic respiration)

During anaerobic conditions, yeast ferments sugar to produce alcohol, carbon

dioxide and water in the process of fermentation.

Equation: Yeast + Glucose = Alcohol + Carbon dioxide + Water

[NOTE: that the same chemistry is used in brewing and baking fermentations, but
brewing uses the alcohol, and baking uses the carbon dioxide.]

Brewing Industry:

Brewer’s yeast (Saccharomyces cerevisiae) ferments sugars in cereal grains to

produce alcohol, in addition to various other products, producing beers and lagers.

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Baking Industry: Baker’s yeast (Saccharomyces cerevisiae) ferments sugars in the
flour, but this time carbon dioxide is the useful product of fermentation. When the
yeast in bread dough releases carbon dioxide it makes bubbles in the dough and
causes the dough to ‘rise’ (increase in volume). The alcohol produced evaporates
during baking.

Myco-protein

The product called Quorn is myco-protein. It is NOT a yeast or a mushroom, but a

filamentous fungus called Fusarium. Mycoprotien is made in fermenters similar to
those found in a brewery. It’s made by adding oxygen, nitrogen, glucose and
minerals to a fungus called Fusarium venenatum.

Myco- protein is used as an alternative to meat in health- and vegetarian products.

It is high in protein, high in fiber, low in saturated fat and contains no cholesterol.

Antibiotics

Antibiotics are used to treat bacterial and fungal infections. You know about
penicillin -produced by the mould Penicillium notatum and discovered by
Alexander Fleming in 1928. Other examples of antibiotics derived from fungi are:
Cephalosporin from Cephalosporium sp. and Griseofulvin from Penicillium
griseofulvum and Penicillium patulum. Today, most antibiotics used in medicine
are derived from bacteria. Antibiotics produced by bacteria include streptomycin
and terramycin.

Statins

Statins are products of metabolic reactions in fungi. Lovastatin comes from

Aspergillus terreus strains; mevastatin from Penicillium citrinum. Statins inhibit an
enzyme involved in the synthesis of cholesterol and they’ve become very
important for control of cholesterol levels in patients. Cholesterol is made in the
liver, but we also get it from our food. Diets high in fat result in a buildup of
cholesterol in the arteries and this can lead to heart attacks or strokes.

Immuno- suppressives

Immune suppressants are essential for organ transplant patients. The T cells of the
human immune system recognise the new organ as ‘foreign’ and begin to destroy

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the organ. The filamentous fungus called Tolypocladium inflatum was found to
produce Cyclosporin A. This drug prevents organ rejection by inhibiting T cell
activation.

Vitamins

All fungi are a good source of vitamins. Brewer’s yeast synthesises B group
vitamins; so, yeast extract and yeast tablets are popular vitamin supplements. In
industry the fungi Nematospora gossypii and Eremothecium ashbyi are now used
to produce B vitamins.

Edible fungi: Certain fungi are edible. About 200 species of mushrooms (Agaricus
sp.) are used as food.
For example: Morels: e.g. Morchella esculenta. Truffles: Underground fruiting
bodies of some Ascomycetes,
Cheese preparation: Pencillium species are used for giving flavour, aroma and
characteristic colour to some cheese.
Soya products: Some species of Aspergillus are used for fermenting and producing
soya sauce and soya paste from soya beans.
synthesis is of organic acids: Citric acid is obtained from some Aspergillus species.
Some fungi yield fumaric acid and lactic acids.
Ergo tine is used to relieve headache by migraine.
Griseofulvin is used to inhibit fungal growth.
Natural dyes: Some natural dyes are obtained from lichens. These are used in
textile industry.

FUNGI IN MEDICINE

The vast majority of the known fungal species are strict saprophytes, although
there are a few capable of causing disease in plants or humans. However, there are
several fungal genera containing species that cause disease (such as infections,
allergies, toxicity) in plants, animals and man. These fungi can be categorized into
two groups in regards to infection:

(1) saprophytic fungi which can be opportunist-tic pathogens that enter via wounds
or due to a weakened state of the host and

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(2) true pathogens that may depend on living plant or human tissues for nutrients
but can also survive outside of the hosts.

Some fungal spe- cies that cause not only disease in plants and but also in humans
include Aspergillus and Fusarium as well as other phytopathogenic fungi now
considered mem- bers of the “emerging pathogen” group in humans.

Aspergillus: The Aspergillus genus are saprophytes found worldwide in soil,

forage products, food products, dust, organic debris and decomposing matter.
Although they are considered weak plant pathogens, there are two species,
Aspergillus flavus and Aspergillus parasiti-cus, which produce potent toxins
(aflatoxins) on certain crops. Especially susceptible to these species are oilseeds
and nuts such as peanuts, corn and cottonseed.

A. flavus is the dominant of the two on corn and cottonseed. In contrast, A.

parasiticus is more prevalent on peanuts than on other crops and, generally,
constitute approxi- mately 10 - 30% of the aflatoxin producing fungi on pea- nuts.

Aflatoxin risk to humans and animals

Death due to aflatoxins has been reported in humans, animals and birds. Aflatoxins
were discovered after groundnut meal contaminated with these toxins killed over
10,000 turkeys (“turkey X disease”) in England [199]. They are potent
heptatotoxic, teratogenic, mutagenic and carcinogenic secondary metabolites that
contaminate crops worldwide, rendering them unsafe for consumption. The four
main aflatoxins are produced by A. flavus, A. parasiticus and Aspergillus nominus

Human infections

Though over 180 Aspergillus species are known, only four are commonly
associated with invasive infection in humans. These species include A. fumigatus,
A. flavus, A. terreus and A. niger. Though saprophytic, they can cause infections in
healthy individuals and, increasingly, are the causative agents of fatal opportunistic
infections in immunocompromised patients. These fungi are ubiquitous worldwide
and produce abundant amounts of asexual, haploid conidia, which are highly
hydrophobic with very small diameters. Their conidia are readily dispersed in air
currents, thereby facilitating exposure. Besides contacting the skin, eyes, and ears,
these conidia are sufficiently small to be inhaled and lodge in all recesses of the

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lung, including the alveoli. The innate immunity of healthy persons protects them
from infection except in certain situations, such as trauma, where conidia may
contaminate the wound.

In cases where the immune system is weakened, the inhaled conidia can germinate
and produce hyphae that invade the surrounding lung tissue, leading to the
development of invasive pulmonary aspergillosis. Invasive aspergillosis is a major
cause of death in immunocompromised patients, with the associated mortality rate
> 90%.

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