MYCOLOGY

VIROLOGY
MLS 302 C - MICROBIOLOGY 3

PRELIMS OUTLINE
General characteristics of fungi Morphology of fungi Classification of fungi Laboratory Diagnosis of Fungal Diseases Classification of Fungal Diseases

REFERENCES
Jawetz, Melnick & Adelbergs Medical
Microbiology

Clinical Mycology by Dismukes, et al.

Bailey & Scotts Diagnostic Microbiology Clinical Microbiology Made Ridiculously Simple

MYCOLOGY

LECTURE 1: OBJECTIVES
1. Define terms used in mycology
2. Discuss the general characteristics of fungi 3. Describe the morphology and structure of fungi in general

4. State the medical importance of the cellular structures of fungi

FUNGAL CHARACTERISTIC
Eukaryotic

Facultative anaerobic/strictly aerobic

Chemotrophic; nutrition by absorption

Non-photosynthetic; achlorophyllos
pH 5-6 (acidic)

High sugar concn. favors growth

 Widely distributed in nature Saprophytic

Gastronomic Delights

Penicillium notatum Tolypocladium inflatum

Phytopathogen

Mycoses

TERMINOLOGY
Mykos (Gk): Mushroom Mycology: Study of fungi

Mycoses: diseases caused by fungi

Dimorphism: ability to grow as yeast at

37C and as mold at room temp. Hyaline fungi: non-pigmented fungi Dematiaceous fungi: pigment-producing fungi

Morphologic Classification of FUNGI

YEAST

MOLD
DIMORPHIC

YEAST
Unicellular fungi MICROSCOPIC Oval to round Budding Bud=Blastospore Pseudohyphae

BUDDING

YEAST

 MACROSCOPIC: Pasty, opaque, cream-colored colonies

 MONOMORPHIC YEAST: Candida albicans

Cryptococcus neoformans Geotrichum candidum

MOLD
Multicellular

Hypha [2-10 m]
MYCELIA Vegetative / Thallus

Reproductive / Aerial

MOLDS: Mycelia

 MACROSCOPIC: Cottony, wooly, velvety, granular, filamentous MONOMORPHIC MOLD: Microsporum Epidermophyton floccosum Trichophyton

 PIGMENTATION: observed from the reverse

 CLASSIFICATION OF HYPHAE
A. EXISTENCE OF SEPTA

In Medically Important Fungi: 3 Types Of Hyphae

1. Coenocytic- sparsely septated
2. Dematiaceous fungi dark and pigmented septate hyphae

3. Hyaline molds: septate, non-pigmented hyphae

B. HYPHAL SHAPES

DIMORPHIC

FUNGI

Mold form: 25C to 30C Yeast form 35C to 37C Thermal dimorphism (a group of pathogenic fungi)

DIMORPHIC FUNGI
Sporothrix schenckii Histoplasma capsulatum Subcutaneous Blastomyces dermatitidis Systemic Coccidioides immitis Paracoccidioides brasiliensis Penicillium marneffei Opportunistic

CAPSULE

Structure: Polysaccharide Functions: Antiphagocytic factor

Cryptococcus neoformans (encapsulated yeast)

CELL WALL
Antigenic Multilayered a. polysaccharides poorly degraded by host activate CF; provokes inflammatory rxn Induce immune hypersensitivity b. proteins and glycoproteins

CELL WALL
Functions: Provides shape, rigidity & strength protection from osmotic shock mediates attachment to host cells

Major Polysaccharides Of Fungal Cell Wall

POLYMER Chitin Chitosan Cellulose -Glucan -Glucan Mannan MONOMER N-acetyl glucosamine D-Glucosamine D-Glucose D-Glucose D-Glucose D-Mannose

The type & amount of polysaccharide vary from one fungal species to another.

CELLULAR MEMBRANE

Bilayered Structure:
1. Phospholipids -phosphatidylcholine

-phosphatidylethanolamine

2.

- ergosterol

Sterols

- zymosterol

CELLULAR MEMBRANE
Functions:
Protects cytoplasm

regulates intake & secretion of solutes facilitates capsule & cell wall synthesis

 CYTOPLASM Nucleus, nuclear membrane, nucleolus, ER, mitochondria, vacuoles

LECTURE 2: OBJECTIVES
Classify fungi
according to methods of reproduction type of spores they produce.

REPRODUCTION
A. SEXUAL REPRODUCTION
opposite gametes mate to form a zygote Sexual spores

3 phases of sexual reproduction: 1. Plasmogamy fusion of opposite but compatible mating types 2. Karyogamy fusion of 2 nuclei. 3. By meiosis, diploid nucleus gives rise to haploid spores

SEXUAL REPRODUCTION

SEXUAL ZYGOSPORE
Enclosed in a thick wall Rhizopus, Mucor

SPORES

ASCOSPORE
Produced in ascus (sac-like); 2-8 spores/ascus Nucleic fusion of morphologically dissimilar cells Histoplasma capsulatum

BASIDIOSPORE formed externally on a base pedestal called Basidium mushroom

ZOOSPORE
Fusion of cells from 2 diff. hyphae

OOSPORES
thick-walled, long-surviving spore
Phytophthora capsici

B. ASEXUAL REPRODUCTION

SPORE GERMINATION
Fruiting body : Principal structure asexual spores

SPORE DISPERSAL

ASEXUAL SPORES
CONIDIOSPORE

Unicellular / multicellular
Produced in a chain at the end of conidiophore Penicillium Aspergillus

A. Arthrospore
fragmented septate hyphae single, slightly thickened cells

Coccidioides
Geotrichum

B. Blastospore

bud

Candida Cryptococcus

C. Chlamydospore
Thick walled formed along the periphery or tip of the hyphae

SPORANGIOSPORE
developed in a sac (sporangium) Attached to a specialized hyphae (sporangiophore)

C. PARASEXUAL REPRODUCTION
Set of events that lead to genetic exchange via mitotic recombination Initiated by formation of a heterokaryon (thallus w/ 2 haploid nuclei of 2 diff. genotypes) for genetic analysis of imperfect fungi

FUNGI-TAXONOMIC CLASSIFICATION
Depends primarily on the type of sexual spore
Phylum Class Order Family Genus Species -mycota -mycetes -ales -ceae

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

ZYGOMYCOTA
sporangium fungi / common molds molds & blights
(Rhizopus stolonifer)

coenocytic rhizoids

ASCOMYCOTA
sac fungi

Reproduction:
Conidiospores

Ascospores
Budding Ascocarp Ascus

BASIDIOMYCOTA
club fungi Some are used as food; others cause crop damage

Seldom reproduce asexually

 Basidiocarp

annulus
basidia

Volva

DEUTEROMYCOTA
Asexual spores: conidia of various types Septate hyphae most are yeast & molds, some dimorphic

LECTURE 3: OBJECTIVES
Discuss the laboratory methods used in the diagnosis of fungal diseases as to: 1. proper collection, handling, transport and disposal of specimen 2. culture media to use 3. methods of identification

SPECIMEN COLLECTION
Aseptic technique Collect spx from actual infxn site Adequate quantity Accurate label; Prompt delivery

CLINICAL SPECIMEN
Blood Bone Marrow CSF
5 ml to BHI broth
Aspirate 0.5ml to BHI Lumbar tap Vol. is >2ml: centri, smear, culture Vol. is <2ml: use uncentri spx.

Direct culture Biphasic culture bottle Membrane filtration Lysis centrifugation

 Sputum
Bronchial washings

Collect sputum in a.m. for 3 days

No 24 hr. collections

Tracheal aspirate
Throat

sterile NSS for washings

+ few yeast & contaminants Use 2 sterile swabs Use tongue depressor if ? candida

Urine

Midstream, clean-catch, catheter

Centri, smear, culture (sediments)

Process in 2hrs or refrigerate >100,000/ml is significant Vaginal & cervical discharges +few to moderate Candida colonies 2 swabs: for KOH & culture

a. Hair

CUTANEOUS May contain contaminants Pluck hair by roots (forceps) Select hair that fluoresce, broken, scaly 70% alc., scrape discolored areas, collect inner infected nail Nail clippings: cut into small pcs. w/KOH Few Candida & contaminants scrape infected area w/KOH

b. Nail

c. Skin