1. INTRODUCTION TO MEDICAL MYCOLOGY
Lecture 14

2. Learning Outcome Able to define terms use in mycology
Can describe basic characteristic of fungi
Able to discuss fungal reproduction
Able to describe mycoses
Able to discuss laboratory examination for fungal infection.

4. Definitions Mycologists--scientists who study fungi
Mycology--scientific discipline dealing with fungi
Mycoses--diseases caused in animals by fungi
Mykos = mycete = fungus

5. I. FUNGI
Diverse group of heterotrophs.
Many are ecologically important saprophytes (consume dead and decaying matter)
Others are parasites.
Most are multicellular, but yeasts are unicellular.
Most are aerobes or facultative anaerobes.
Cell walls are made up of chitin (polysaccharide).
Over 100,000 fungal species identified. Only about 100 are human or animal pathogens.
Most human fungal infections are nosocomial and/or occur in immunocompromised individuals (opportunistic infections).
Fungal diseases in plants cause over 1 billion dollars/year in losses.

6. Characteristic of fungi
Vegetative Hypha
Composed of cells involved in catabolism and growth.
Reproductive Hypha (aerial)
Composed of cells involved in reproduction (produce spores).

7. General knowledge of the fungi
Both sexual and asexual spore may be produced
Store their food as glycogen (plant; starch)
Fungi are heterotrophic organisms, lack of chlorophyll (plant; autotrophic)

8. General knowledge of the fungi
Yeast : unicellular, 370C
Budding Yeast – may produce a pseudohypa
Fission yeast
Mold : multicellular, hyphae, 250C
Dimorphic fungi (thermally dimorphic fungi) : mold phase & yeast phase

9. Yeasts Facultative Anaerobes Fermentation : ethanol and CO2
Non-filamentous unicellular fungi
Spherical or oval
Reproduction:
a) by fission, or
b). By budding

10. Yeast Reproduction FISSION
“even” reproduction, nucleus divides forming two identical cells, like bacteria
BUDDING
“uneven” reproduction, parent cell’s nucleus divides and migrates to form a bud and then breaks away

11. YEAST Unicellular Micr.: Oval to round (Diameter : 3-15 µm)
Macr.: Pasty colonies (resemble bacteria)

12. MOULD Multicellular Micr.: . Hypha(e) (dia: 2-10 µm)
. Spores / conidia.
Macr.: .Surface texture: Cottony/ powdery/ wooly/velvety/granular/glabrous
Pigmentation :obverse & reverse

13. General knowledge of the fungi
Eukaryotic microorganisms
Rigid cell walls: chitin, glucans, mannans
Plasma membranes: ergosterol
Lysine synthesis by L- amino adipic acid (AAA) pathway and other organisms synthesize lysine by diaminopimelic acid (DAP) pathway

14. Fungal Structure Thallus-”body” Mycelium –
Molds & fleshy fungi have these structures
Long filaments of cells (hyphae):
Septate hyphae (cross wall) :most fungi
Aseptate hyphae (coenocytic ) :no cross wall, continous mass with many nuclei .
Mycelium –
Abundance growth of aerial hyphae resulting a mass can be observed with unaided ayes

17. Fungal Classification
Four groups of true fungi
Zygomycetes (common bread mold—Rhizopus)
Basidiomycetes (puffballs & common mushrooms)
Ascomycetes (Dutch elm disease/rye smut)
Deuteromycetes (fungi imperfection)

18. Classification ( con’t) :
First three groups is based on their method of sexual reproduction
4th group, the Deuteromycetes, have NO sexual reproduction

19. Fungi-Taxonomic classification
SEXUAL SPORE CLASS
Zygospore Zygomycetes
Basidiospore Basidiomycetes
Ascospore Ascomycetes
None/Unknown---- Deuteromycetes
(“Fungi Imperfecti”)

20. Zygomycetes Asexual phase—Sporangium—bread mold (Rhizopus stolonifer)
Sexual phase--- sporgangium ---shotgun fungus (lives on dung) it shoots its sporgangium explosively towards light or fly pathogen (Entomophthora muscae—--these types of fungi have been used as agents for biological control of insects

21. Basidiomycetes Basidiospore
Examples: boletes, puffballs,smuts, stinkhorns and tooth fungi

22. Ascomycetes Asexual phase- Sexual phase (morels, lichens )
Conidiospores (Penicillium and Aspergillus)
budding yeast
Sexual phase (morels, lichens )

23. Life Cycle Fungal reproduce by 2 way Asexual Sexual
Hyphae fragmentation
Asexual spores
Conidiosphore
Arthroconidia
Blastoconidia
sporangiospore

24. Reproduction of Fungi 1. Sexual reproduction --Sexual spores
2. Asexual reproduction--Asexual spores
3. Parasexual reproduction--Genetic exchange

25. Life cycle
Conidiosphore
Spore that is not enclosed in sac

26. Life cycle Athroconidia
Form fragmentation of hypae into single thick cells.

27. Life cycle Blastoconidia Buds coming from parents cells
Chlamydoconium a variant which spore form by thickening and enlargement within hypae

28. Life cycle Sporangiospore
Spore form inside sporangium (sac) at the end of sporangiophore

29. Asexual Spores Conidiospores Chlamydospores Sporangiospores
Blastospores
Arthrospore

30. Asexual spores ( con’t)
Conidiospore
Multiple (chains) or single spores formed at the end of an aerial hypha
Not enclosed within a sac
Aspergillus spp.
Penicillium spp

32. Aspergillus sp.

34. Conidial Fungus reproduces by means of asexual spores called conidia
Conidia vary greatly in shape, size and color
Most of the common household molds & mildews are conidial fungi

35. Asexual spores ( cont) Another type of Conidiophore: Blastospores
A bud coming off the parent cell
Candida albicans

36. Chlamydospore
Formed within hypha
Thick-walled spore
Candida albicans

37. Chlamydospores
The chlamydospore is a method of producing a substantial resting spore very quickly
Nutrient is shunted from adjacent cells into a preferred cell and it swells up, converts nutrient materials to oil droplets for efficient storage, then rounds off with a thick, often roughened outer wall for protection

38. Asexual spores Sporangiospores
Hundreds formed within a sac (sporangium) at the end of an aerial hypha
Rhizopus spp.
Mucor spp
Absidia spp

40. Types of asexual spores

42. SEXUAL Spores
1. Zygospore
2. Ascospore
3. Basidiospore
4. Oospore

43. Sexual spores Have three stages
Plasmogamy- a haploid nucleus of donor cells
Karyogamy- the (+) and (-) nuclei fuse to form a diploid zygote
Meiosis – the diploid nucleus give rise to haploid nuclei (sexual spores) genetic recombinant

47. MYCOSES Superficial ( skin, hair, cornea)
Cutaneous ( Dermatophytosis )
Subcutaneous
True systemic (endemic)
Opportunistic

48. Zygomycosis
Disease caused by fungi that are classifiable as Zygomycetes
Mucormycosis : Order Mucorales
Entomophthoromycosis : Order Entomophthorales

49. Zygomycosis (cn’t)
Presents as a spectrum of diseases, depending on the portal of entry and the predisposing risk factors of the patient:
Rhinocerebral zygomycosis
Pulmonary zygomycosis
Gastrointestinal zygomycosis
Cutaneous zygomycosis
Disseminated zygomycosis

50. Material from the periorbital tissue of a woman with poorly controlled diabetes mellitus with facial and periorbital swelling due to zygomycosis (see right picture) is stained with periodic acid-Schiff stain (X 560). The material demonstrates the classic appearance of irregularly shaped broad hyphae with right-angle branching (arrow).

51. Laboratory to diagnosis of fungal infection
Specimen collection and transport
Specimen processing
Direct examination
Selection and inoculation of culture media
Identification

52. LABORATORY DIAGNOSIS OF MYCOSES
Direct microscopic examination
Culture
Serology

53. Specimen collection and transport
must be material from the actual infection site
must be carefully the contamination
must be established for the best chance of recovery of causative microorganisms (optimal times)

54. Specimen collection and transport ( con’t)
must be obtained to perform the culture or other techniques request (sufficient quantity)
must be used to ensure optimal recovery of microorganisms
obtain cultures before the treatment
the culture container must be properly labeled

55. Specimen processing specimen should be examined as soon as possible
direct examination :
KOH mount
Calcofluor white
India ink
culture media

56. Selection and inoculation of culture media
Culture media for recovery of fungi from clinical specimens. PDA, Corn Starch Agar
The recovery rate may be somewhat enhanced by using a variety of isolation media, considerations of cost, storage, incubator space and technologist time.

57. Initial observations in the study of fungus isolates
1.Appearance of the growth
2. Rate of growth
3. Colony pigmentation
4. Growth on media containing antifungal agents
5.Dimorphic fungi

58. Initial observations in the study of fungus isolates
1.Appearance of the growth
- surface and reverse surface of colony were observed
- delicate or hairlike hyphae
2. Rate of growth
- saprophytes : 3-5 days
- dimorphic fungi : 10 days or more
- dermatophytes : 14 days or more

59. Initial observations in the study of fungus isolates
3. Colony pigmentation
4. Growth on media containing antifungal agents
- most strains of the dimorphic fungi can grow
- most strains of the rapidly growing saprobes are
inhibited
5. Dimorphic growth
- mold form (the environmental and infective form) ;
ambient or room temperature (22-25 OC)
- yeast form (invasive form) ;
near body temperature (30-35 OC)

60. Preparation of mounts for study
The tease mount
Scotch tape preparation
The microslide culture technique
( slide culture )

61. Terms useful in the examination of fungi
hypha and pseudohyphae
mycelium
septate or aseptate (or coenocytic) hyphae
vegetative mycelium
aerial mycelium
reproductive mycelium

65. Exercise List 4 classes of fungi List types of asexual spores
Diferentiate between conidiospore with sporangiospores