INTRODUCTION TO MEDICAL MYCOLOGY

Departemen Mikrobiologi FK USU, Medan

Specific learning objectives:

Menjelaskan dasar-dasar biologi jamur

dan jamur patogen sistemik / oportunistik pada manusia berdasarkan klasifikasi dan morfologinya Menjelaskan cara diagnosis jamur

Pendahuluan

Definitions
Mycologists--scientists who study fungi
Mycology--scientific discipline dealing

with fungi Mycoses--diseases caused by fungi in human or animals Mykos = mycete = fungus

Mycology is the study of fungi, which are organism that contain true nuclei (eukaryotic), are devoid of chlorophyll, and absorb all nutrients from the environment, especially from decaying organic matter
5

Fungal Structure
Thallus-body

Molds & fleshy fungi have these structures Long filaments of cells (hyphae) Septate hypha :Cross Wall (Most Fungi) Aseptate hypha (Coenocytic) -No cross
wall, continuous mass with many nuclei

General knowledge of the fungi

Eukaryotic microorganisms
Rigid cell walls: chitin, glucans, mannans

and peptidoglycans Plasma membranes: ergosterol Lysine synthesis by L- amino adipic acid (AAA) pathway and other organisms synthesize lysine by diaminopimelic acid (DAP) pathway
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)

General knowledge of the fungi

Yeast : unicellular, 370C
Mold : multicellular, hyphae, 250C Dimorphic fungi (thermally dimorphic

fungi) : moldyeast

Dimorphic Fungi
Capable of growing in mould or

yeast form under different environmental conditions (temperature, CO2, nutrients) of pathogenic fungi) e.g. :

Thermal dimorphism (a group

10

Thermally dimorphic fungi :

Blastomyces

dermatitidis Coccidioides immitis ( C.posadasii ) Histoplasma capsulatum

Var.capsulatus Var.duboisii

Paracoccidioides

brasiliensis Penicillium marneffei

11

Fungi- Morphological Classification

Yeast Mould

Dimorphic

12

YEAST
Unicellular

Micr.: Oval to round

(Diameter : 3-15 m)

Macr.: Pasty colonies (resemble bacteria)

13

Yeasts
Facultative Anaerobes Fermentation : ethanol and CO2 Non-filamentous unicellular fungi

Spherical or oval yeast cells

Reproduction of Yeasts: a by Fission ,or . b) by Budding

14

Yeast Reproduction
FISSION

even reproduction, nucleus divides forming two identical cells, like bacteria BUDDING uneven reproduction, parent cells nucleus divides and migrates to form a bud and then breaks away

15

MOULD
Multicellular

Micr.: Hypha(e) (dia: 2-10 m) Asexual Spores Macr.: Surface texture: Cottony/ hairy wooly/ velvety/ granular/glabrous Pigmentation: observed from the reverse
16

MOULD
Hypha
Mycelium: a. Vegetative

b. Aerial / Fertile / Reproductive

17

Classification of Hyphae
A. Existence of septa: o Septate o Nonseptate ( coenocytic ) B. Shape and Morphology Racquet Spiral Nodular Root-like (rhizoid) Pectinate Favic chandelier
18

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

19

Classification ( cont) :
First three groups is based on their

method of sexual reproduction

4th group, the Deuteromycetes, have NO

sexual reproduction

20

Reproduction of Fungi
1. Sexual reproduction --Sexual spores

2. Asexual reproduction--Asexual spores 3. Parasexual reproduction--Genetic exchange

21

Types of spores
Spora aseksual:

The product of mitotic division of a single parent cell Formed through a process involving the fusing of two parental nuclei followed by meiosis

Spora seksual:

22

Asexual spore formation

Berdasarkan kepada nature of the

reproductive hyphae & the manner in which the spores originate, terdapat dua subtipe spora aseksual Progeny will be identical to parents

Sporangiospores Conidium ( pl. conidia )

23

Spora aseksual ( cont)

Sporangiospores adalah spora-spora

yang dibentuk dengan adanya successive cleavage di dalam sac-like head yang dinamai sporangium yang terdapat di ujung sporangiophora. Spora-spora ini mulanya terkurung di dalam sporangium dan akan dibebaskan jika dinding sporangium rupture.
24

 Konidia adalah free spores yang tidak

terdapat di dalam spore-bearing sac. Konidia ini berkedmbang dengan cara:

The pinching of the tip of a special hyphae, or The segmentation of a pre-existing vegetative hyphae.

25

Asexual reproduction
Tipe-tipe Conidia :

Arthrospore Blastospores, Chlamydospores Porospore Phialospore Macro & microconidia

26

Examination of asexual spores

Fungal spores are different from the

endospores of bacteria Endospores of bacteria are not for reproduction, but an environmentally resistant life stage Fungal spores are for reproduction, do not ensure resistance to environmental conditions
27

Asexual spore :Conidia

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

28

Asexual Spores
Conidia

Chlamydospores Blastospores Arthrospore Porospore Phialospore Macro- microconidia

29

Tipe-tipe Konidia
Arthrospore ( Gr.arthron = joint)

Spora-spora yang berbentuk empat persegi panjang yang dihasilkan jika terjadi fragmentasi septate hyphae di cross wall Contoh : Coccidioides immitis

30

 Chlamydospores ( Gr. Chlamys =cloack)

Konidia yang berbentuk spheris yang terbventuk jika sel-sel hifa menebal dan akan dibebaskan jika hifa disekeliling nya ruptur Spora-spora ini bertindak sebagai survival cell atau resting cell Contoh.: Candida albicans
31

 Blastospore ( blasto = a bud )

Spora-spora yang dibentuk secara budding ( pertunasan) dari sel induknya, Contoh : Candida sp.

32

 Phialophora ( Gr.phialos = a vase ).

Konidium yang budded dari mulut a vase-shaped spore-bearing cell yang dinamai phialide (sterigma) yang akan meninggalkan a small collar (kerah). Contoh : Aspergillus, Penicillium

33

 Porospore

Adalah konidium yang grows out melalui pori-pori dari spore-beanting cells Contoh :

34

 Micro- & macroconidium

Konidia yang ukurannya lebih kecil dan lebih besar yang dihasilkan oleh jamur yang sama pada berbagai kondisi Microconidium : satu sel Macroconidium: dua sel atau lebih Perhatikan dinding selnya apakah smooth atau spiny.

35

36

Asexual spores ( cont)

Conidia ( Phialoconidia )

Multiple (chains) or single spores formed at the end of an aerial hypha Not enclosed within a sac
Aspergillus

spp. Penicillium spp

37

Aspergillus flavus

38

Asexual spores ( cont)

Blastospores
A

bud coming off the parent cell

albicans

Candida

39

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
40

 Chlamydospore

Formed within hypha ( intercalary ) or on the tip hyphae ( terminal ) Thick-walled spore Resting spore or survival.
Candida

albicans

41

C.Albicans : chlamydospores

42

Asexual spores ( cont )

Sporangiospores

Hundreds formed within a sac (sporangium) at the end of an aerial hypha (sporangiophore ) ,e.g. :

Rhizopus spp. Mucor spp Absidia spp

43

Sporangium & Sporangiospores

44

SUMMARY
Reproduction of Fungi
1. Sexual reproduction --Sexual spores 2. Asexual reproduction--Asexual spores 3. Parasexual reproduction--Genetic exchange
45

Types of asexual spores

46

SEXUAL Spores
1. Zygospore
2. Ascospore 3. Basidiospore 4. Oospore
47

Asexual Spores
1. Arthrospore 2. Blastospore 3. Chlamydospore 4. Phialospore 5 .Porospore 6. Conidium ( pl conidia ):

macroconidia microconidia

48

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

49

Laboratory to diagnosis of fungal infection

Specimen collection and transport
Specimen processing Direct examination Selection and inoculation of culture media Identification

50

LABORATORY DIAGNOSIS OF MYCOSES

Direct microscopic examination
Culture : macroscopic & microscopic

obsevations
Serology

51

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)

52

Specimen collection and transport cont)

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

53

Specimen processing

specimen should be examined as soon as possible direct examination :

KOH mount Calcofluor white KOH + Calcofluor white India ink

culture media

54

Selection and inoculation of culture media

Culture media for recovery of fungi from clinical specimens need not be elaborate. The recovery rate may be somewhat enhanced by using a variety of isolation media, considerations of cost, storage, incubator space and technologist time.

55

Initial observations in the study of fungus isolates

1.Appearance of the growth 2. Rate of growth 3. Colony pigmentation ( surface & reverse ) 4. Growth on media containing antifungal agents 5.Dimorphic fungi

56

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
57

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)
58

Preparation of mounts for study

The tease mount Scotch tape preparation The microslide culture technique ( slide culture )

59

Terms useful in the examination of fungi

hypha and pseudohyphae mycelium septate or aseptate (or coenocytic) hyphae vegetative mycelium aerial mycelium reproductive mycelium

60

Types of asexual spores

61

Fungal Reproduction
fungi replicate by mitosis rather than the

binary fission employed by bacteria. Types of fungal reproduction, :

budding fission hyphae fragmentation sporulation

62

Sexual & Asexual Spores

Fungal spores come in two varieties: asexual spores and sexual spores Spores are used extensively to identify fungi.
Asexual spores

Asexual spores are formed by a single parental fungi and therefore genetically identical to the parental fungi. Asexual spores come in a variety of types formed by a variety of mechanisms including:

63

Sexual & Asexual Spores

Asexual spores

Asexual spores are formed by a single parental fungi and therefore genetically identical to the parental fungi. Asexual spores come in a variety of types formed by a variety of mechanisms including:

Arthrospores (sliced bread pieces) Blastospores (buds on a twig) Chlamydospores (giant cell with oil) Conidiospores or conidia (fingers) Sporangiospores (sac) Phialospores ( phialides) Porospores ( pores).
64

65

Ascomycetes
Asexual phase Conidiospores

(Penicillium and Aspergillus)

budding yeast

Sexual phase (morels, lichens )

66

Zygomycetes
Asexual phaseSporangiumbread mold

(Rhizopus stolonifer) Sexual phase--- sporangium ---shotgun fungus (lives on dung) it shoots its sporangium explosively towards light or fly pathogen (Entomophthora muscae--these types of fungi have been used as agents for biological control of insects

67

Basidiomycetes
Basidiospore
Examples: boletes, puffballs,smuts,

stinkhorns and tooth fungi

68

69

70

Mycology not Micturation

71

Mycology

makes Fun gi

72