Basic Mycology

(Eumycota)
By:Dr.Tahseen Ismail
Introduction:
• Microbiologists use the term fungus [pl., fungi; Latin fungus, mushroom] to describe eucaryotic
organisms that are spore-bearing, have absorptive nutrition, lack chlorophyll, and reproduce
sexually and asexually.
• Scientists who study fungi are mycologists [Greek mykes, mushroom, and logos, science], and the
scientific discipline devoted to fungi is called mycology.
• The study of fungal toxins and their effects is called mycotoxicology, and the diseases caused by
fungi in animals are known as mycoses (s., mycosis). According to the universal phylogenetic tree,
fungi are members of the domain Eucarya.
• Fungi are widely distributed and are found wherever moisture is present. They are of great
importance to humans in both beneficial and harmful ways.
• The fungi have a long and confused taxonomic history. Their relatively simple morphology, wide
diversity, and lack of a fossil Fungi.
• Like protists, fungi record limit the value of traditional taxonomic approaches. Presently eight
fungal groups:
• Chytridiomycetes, Zygomycota, Ascomycota, Basidiomycota, Urediniomycetes,Ustilaginomycetes,
Glomeromycota, and Microsporidia.
• Morphological, biochemical and molecular phylogenetic analyses demonstrate that the Fungi
constitute a monophyletic group. They are sometimes referred to as the true fungi or Eumycota
Distribution & Importance

• Fungi are primarily terrestrial organisms, although a few are freshwater or marine. They have a
global distribution from polar to tropical regions.
• Many are pathogenic and infect plants and animals. Fungi also form beneficial relationships with
other organisms. For example, the vast majority of vascular plant roots form associations (called
mycorrhizae) with fungi.
• Fungi also are found in the upper portions of many plants. These endophytic fungi affect plant
reproduction and palatability to herbivores.
• Lichens are associations of fungi and photosynthetic protists or cyanobacteria.
• About 90,000 fungal species have been described; however, some estimates suggest that 1.5
million species may exist.
• Fungi are important to humans in both beneficial and harmful ways. With bacteria and a few other
groups of chemoorganotrophic organisms, fungi act as decomposers, a role of enormous
significance.
• They degrade complex organic materials in the environment to simple organic compounds and
inorganic molecules. In this way carbon, nitrogen, phosphorus, and other critical constituents of
dead organisms are released and made available for living organisms.
• Fungi can be a major cause of disease. Plants are particularly vulnerable to fungal diseases
because fungi can invade leaves through their stomates.
• Over 5,000 species attack economically valuable crops, garden plants, and many wild plants.
• Fungi also cause many diseases of animals and humans. In fact, about 20 new human fungal
pathogens are documented each year.
• Fungi, especially the yeasts, are essential to many industrial processes involving fermentation.
Examples include the making of bread, wine, and beer.
• Fungi also play a major role in the preparation of some cheeses, soy sauce, and sufu; in the
commercial production of many organic acids (citric, gallic) and certain drugs (ergometrine,
cortisone); and in the manufacture of many antibiotics (penicillin, griseofulvin) and the
immunosuppressive drug cyclosporin.
• Finally, fungi are important research tools in the study of fundamental biological processes.
• Cytologists, geneticists, biochemists, biophysicists, and microbiologists regularly use fungi in their
research.
• The yeast Saccharomyces cerevisiae is the best understood eucaryotic cell. It has been a valuable
model organism in the study of cell biology, genetics, and cancer.
Fungal Pathogens of Plants
STRUCTURE
• The body or vegetative structure of a fungus is called a thallus [pl., thalli]. It varies in complexity and size,
ranging from the single-cell microscopic yeasts to multicellular molds, macroscopic puffballs,
and mushrooms.
• The fungal cell usually is encased in a cell wall of chitin.
• Chitin is a strong but flexible nitrogen containing polysaccharide consisting of N-acetylglucosamine residues.
• There are two types of fungi: yeasts and molds.
• Yeasts grow as single cells that reproduce by asexual budding. Molds grow as long filaments (hyphae) and
form a mat (mycelium).
• A yeast is a unicellular fungus that has a single nucleus and reproduces either asexually by budding and
transverse division or sexually through spore formation. Each bud that separates can grow into a new yeast,
and some group together to form colonies.
• Generally yeast cells are larger than bacteria, vary considerably in size, and are commonly spherical to egg
shaped.
• They lack flagella but possess most of the other eucaryotic organelles.
• The thallus of a mold consists of long, branched, threadlike filaments of cells called hyphae [s., hypha; Greek
hyphe, web] that form a mycelium (pl., mycelia), a tangled mass or tissue like aggregation of hyphae In some
fungi, protoplasm streams through hyphae, uninterrupted by cross walls These hyphae are called coenocytic
or aseptate.
• The hyphae of other fungi have cross walls called septa (s., septum) with either a single pore or multiple
pores that enable cytoplasmic streaming. These hyphae are termed septate.
Fungal Thalli.
A Yeast. Diagrammatic drawing of a yeast cell showing typical morphology
• Hyphae are composed of an outer cell wall and an inner lumen, which contains the cytosol and
organelles.
• A plasma membrane surrounds the cytoplasm and lies next to the cell wall. The filamentous nature
of hyphae results in a large surface area relative to the volume of cytoplasm. This makes adequate
nutrient absorption possible.
• The fungal cell membrane contains ergosterol, in contrast to the human cell membrane, which
contains cholesterol. The selective action of amphotericin B and azole drugs, such as fluconazole
and ketoconazole, on fungi is based on this difference in membrane sterols.
YM shift
• Several medically important fungi are thermally dimorphic (i.e., they form different structures at different
temperatures).
• They exist as molds in the environment at ambient temperature and as yeasts (or other structures) in human
tissues at body temperature.
• Dimorphic fungi can change from the yeast (Y) form in the animal to the mold or mycelial form (M) in the
external environment in response to changes in various environmental factors (nutrients, CO2 tension,
oxidation-reduction potentials, temperature). This shift is called the YM shift.
• In plant-associated fungi the opposite type of dimorphism exists: the mycelial form occurs in the plant and the
yeast form in the external environment.
Nutrition and Metabolism:
• Fungi grow best in dark, moist habitats where there is little danger of desiccation, but they are
found wherever organic material is available.
• Most fungi are saprophytes, securing their nutrients from dead organic material.
• Like many bacteria and protists, fungi release hydrolytic exoenzymes that digest external
substrates. They
then absorb the soluble products—a process sometimes called osmotrophy.
• They are chemo-organoheterotrophs and use organic compounds as a source of carbon, electrons,
and energy
• Glycogen is the primary storage polysaccharide in fungi. Most fungi use carbohydrates (preferably
glucose or maltose) and nitrogenous compounds to synthesize their own amino acids and proteins.
• Fungi usually are aerobic. Some yeasts, however, are facultatively anaerobic and can obtain energy
by fermentation.
• Many fungal fermentations are of industrial importance, such as the production of ethyl alcohol in
the manufacture of beer and wine.
• Obligately anaerobic fungi are found in the rumen of cattle.
REPRODUCTION

• Reproduction in fungi can be either asexual or sexual. Asexual reproduction is accomplished in

several ways:
1. A parent cell can undergo mitosis and divide into two daughter cells by a central constriction and
formation of a new cell wall.
2. Mitosis in vegetative cells may be concurrent with budding to produce a daughter cell. This is
very common in the yeasts.
3. The most common method of asexual reproduction is spore production. Asexual spore formation
occurs in an individual fungus through mitosis and subsequent cell division.
• There are several types of asexual spores, each with its own name:
• a. A hypha can fragment (by the separation of hyphae through splitting of the cell wall or septum)
to form cells that behave as spores. These cells are called arthroconidia or arthrospores.
• b. If the cells are surrounded by a thick wall before separation, they are called chlamydospores.
• c. If the spores develop within a sac (sporangium; pl., sporangia) at a hyphal tip, they are called
sporangiospores.
• d. If the spores are not enclosed in a sac but produced at the tips or sides of the hypha, they are
termed conidiospores.
• e. Spores produced from a vegetative mother cell by budding are called blastospores.
(a) Transverse fission. (b) Hyphal fragmentation resulting in arthroconidia (arthrospores) and (c) chlamydospores.
(d) Sporangiospores in a sporangium. (e) Conidiospores arranged in chains at the end of a conidiophore.
(f) Blastospores are formed from buds off of the parent cell
• Sexual reproduction in fungi involves the fusion of compatible nuclei. Homothallic fungal species
are self-fertilizing and produce sexually compatible gametes on the same mycelium.
• Sexual reproduction must occur between mycelia of opposite mating types (MAT). However, one
instance of same-sex mating was discovered following an outbreak of the pathogenic yeast
Crytococcus gatti in Canada.
• Depending on the species, sexual fusion may occur between haploid gametes, gamete-producing
bodies called gametangia, or hyphae.
• Sometimes both the cytoplasm and haploid nuclei fuse immediately to produce the diploid zygote.
Usually, however, there is a delay between cytoplasmic and nuclear fusion.
• This produces a dikaryotic stage in which cells contain two separate haploid nuclei (N N), one
from each parent.
• After a period of dikaryotic existence, the two nuclei fuse and undergo meiosis to yield spores.
• For example, in the zygomycetes the zygote develops into a zygospore; in the ascomycetes, an
ascospore ; and in the basidomycetes; a basidiospore
Reproduction in Fungi.
• The diagram shows generalized life cycle for fungi showing the alternation of
haploid and diploid stages. Some fungal species do not pass through the dikaryotic
stage indicated in this drawing.
• The asexual (haploid) stage is used to produce spores that aid in the dissemination
of the species.
• The sexual (diploid) stage involves the formation of spores that survive adverse
environmental conditions (e.g., cold, dryness, heat)
Diagrammatic representation of the life cycle of Rhizopus stolonifer.
Both the sexual and asexual phases are illustrated
Importance of fungal spores
• Fungal spores are important for several reasons.
• The spores enable fungi to survive environmental stresses such as desiccation, nutrient limitation,
and extreme temperatures, although they are not as stress resistant as bacterial endospores.
• Because they are often small and light, spores can remain suspended in air for long periods. Thus
they frequently aid in fungal dissemination, a significant factor that helps explain the wide
distribution of many fungi.
• Fungal spores often spread by adhering to the bodies of insects and other animals.
• The bright colors and fluffy textures of many molds often are due to their aerial hyphae and
spores.
• The size, shape, color, and number of spores are useful in the identification of fungal species.
PATHOGENESIS

• The response to infection with many fungi is the formation of granulomas.

• Granulomas are produced in the major systemic fungal diseases (e.g., coccidioidomycosis,
histoplasmosis, and blastomycosis, as well as several others).
• The cell-mediated immune response is involved in granuloma formation.
• Acute suppuration, characterized by the presence of neutrophils in the exudate, also occurs in
certain fungal diseases such as aspergillosis and sporotrichosis.
• Fungi do not have endotoxin in their cell walls and do not produce bacterial-type exotoxins.
• Activation of the cell-mediated immune system results in a delayed hypersensitivity skin test
response to certain fungal antigens injected intradermally.
• A positive skin test indicates exposure to the fungal antigen. It does not imply current infection,
because the exposure may have occurred in the past.
• A negative skin test makes the diagnosis unlikely unless the patient is immunocompromised.
• Because most people carry Candida as part of the normal flora, skin testing with Candida antigens
can be used to determine whether cell-mediated immunity is normal.
• Intact skin is an effective host defense against certain fungi (e.g., Candida, dermatophytes), but if the skin is
damaged, organisms can become established.
• Fatty acids in the skin inhibit dermatophyte growth, and hormone-associated skin changes at puberty limit ringworm
of the scalp caused by Trichophyton.
• Normal flora of the skin and mucous membranes suppress fungi. When the normal flora is inhibited (e.g., by
antibiotics), overgrowth of fungi such as C. albicans can occur.
• In the respiratory tract, the important host defenses are the mucous membranes of the nasopharynx, which trap
inhaled fungal spores, and alveolar macrophages.
• Circulating IgG and IgM are produced in response to fungal infection, but their role in protection from disease is
uncertain.
• The cell-mediated immune response is protective; its suppression can lead to reactivation and dissemination of
asymptomatic fungal infections and to disease caused by opportunistic fungi.
FUNGAL TOXINS & ALLERGIES
• In addition to mycotic infections, there are two other kinds of fungal disease:
• (1) mycotoxicoses, caused by ingested toxins, and
• (2) allergies to fungal spores.
• The best-known mycotoxicosis occurs after eating Amanita mushrooms. These fungi produce five
toxins, two of which—amanitin and phalloidin—are among the most potent hepatotoxins.
• The toxicity of amanitin is based on its ability to inhibit cellular RNA polymerase, which prevents
mRNA synthesis.
• Another mycotoxicosis, ergotism, is caused by the mold Claviceps purpurea, which infects grains
and produces alkaloids (e.g., ergotamine and lysergic acid diethylamide [LSD]) that cause
pronounced vascular and neurologic effects.
• Other ingested toxins, aflatoxins, are coumarin derivatives produced by Aspergillus flavus that
cause liver damage and tumors in animals and are suspected of causing hepatic carcinoma in
humans.
• Aflatoxins are ingested with spoiled grains and peanuts and are metabolized by the liver to the
epoxide, a potent carcinogen.
• Aflatoxin B1 induces a mutation in the p53 tumor suppressor gene, leading to a loss of p53 protein
and a consequent loss of growth control in the hepatocyte.
• Allergies to fungal spores, particularly those of Aspergillus, are manifested primarily by an
asthmatic reaction (rapid bronchoconstriction mediated by IgE), eosinophilia, and a “wheal and
flare” skin test reaction.
• These clinical findings are caused by an immediate hypersensitivity response to the fungal spores.
LABORATORY DIAGNOSIS:
• There are four approaches to the laboratory diagnosis of fungal diseases:
• (1) direct microscopic examination, (2) culture of the organism, (3) DNA probe tests, and (4)
serologic tests
• Direct microscopic examination:
• Direct microscopic examination of clinical specimens such as sputum, lung biopsy material, and
skin scrapings depends on finding characteristic asexual spores, hyphae, or yeasts in the light
microscope.
• The specimen is either treated with 10% potassium hydroxide (KOH) to dissolve tissue material,
leaving the alkali-resistant fungi intact, or stained with special fungal stains.
• Some examples of diagnostically important findings made by direct examination are:
• (1) the spherules of C. immitis and (2) the wide capsule of Cryptococcus neoformans seen in India
ink preparations of spinal fluid. Calcofluor white is a fluorescent dye that binds to fungal cell walls
and is useful in the identification of fungi in tissue specimens.
• Methenamine silver stain is also useful in the microscopic diagnosis of fungi in tissue.
• Some examples of diagnostically important findings made by direct examination are:
• (1) the spherules of C. immitis and (2) the wide capsule of Cryptococcus neoformans seen in India
ink preparations of spinal fluid. Calcofluor white is a fluorescent dye that binds to fungal cell walls
and is useful in the identification of fungi in tissue specimens.
• Methenamine silver stain is also useful in the microscopic diagnosis of fungi in tissue
• (2) culture of the organism:
• Fungi are frequently cultured on Sabouraud’s agar, which facilitates the appearance of the slow-
growing fungi by inhibiting the growth of bacteria in the specimen
• Inhibition of bacterial growth is due to the low pH of the medium and to the chloramphenicol and
cycloheximide that are frequently added. The appearance of the mycelium and the nature of the
asexual spores are frequently sufficient to identify the organism.
• (3) DNA probe tests:
• Tests involving DNA probes can identify colonies growing in culture at an earlier stage of growth than can tests
based on visual detection of the colonies. As a result, the diagnosis can be made more rapidly. At present, DNA
probe tests are available for Coccidioides, Histoplasma, Blastomyces, and Cryptococcus.
• (4) Serologic tests:
• Tests for the presence of antibodies in the patient’s serum or spinal fluid are useful in diagnosing systemic mycoses
but less so in diagnosing other fungal infections. As is the case for bacterial and viral serologic testing, a significant
rise in the antibody titer must be observed to confirm a diagnosis.
• The complement fixation test is most frequently used in suspected cases of coccidioidomycosis, histoplasmosis, and
blastomycosis.
• In cryptococcal meningitis, the presence of the polysaccharide capsular antigens of C. neoformans in the spinal fluid
can be detected by the latex agglutination test
ANTIFUNGAL THERAPY

• The drugs used to treat bacterial diseases have no effect on fungal diseases.
• For example, penicillins and aminoglycosides inhibit the growth of many bacteria but do not affect the growth of
fungi.
• This difference is explained by the presence of certain structures in bacteria (e.g., peptidoglycan and 70S ribosomes)
that are absent in fungi.
• The most effective antifungal drugs, amphotericin B and the various azoles, exploit the presence of ergosterol in
fungal cell membranes that is not found in bacterial or human cell membranes.
• Amphotericin B (Fungizone) disrupts fungal cell membranes at the site of ergosterol and azole drugs inhibit the
synthesis of ergosterol, which is an essential component of fungal membranes.
• Another antifungal drug, caspofungin (Cancidas), inhibits the synthesis of β-glucan, which is found in fungal cell
walls but not in bacterial cell walls. Human cells do not have a cell wall.
•