MEDICAL MYCOLOGY. Medical Mycology Reference Laboratory
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1 MEDICAL MYCOLOGY Medical Mycology Reference Laboratory Laboratory of Parasitology and Mycology Institute of Microbiology and Immunology Medical Faculty in Belgrade, 2010
2 The goal of teaching: 1. The morphology and biology of the fungi Classification of medically important fungi Fungal morphology and structure Antifungals - Mechanisms of action Fungi - biology and physiology Fungal infections (FI) - classification 3. Common fungi which causes superficial / invasive / oprtunistic FI Candida - morphology, biology Pathogenesis Cryptococcus - morphology, biology. Pathogenesis Aspergillus - morphology, biology. Pathogenesis Fusarium - morphology, biology. Pathogenesis 2. Common fungi which primarily cause skin, hair and nail FI Malassezia - morphology, biology Pathogenesis of pitiriasis versicolor, Pathogenesis of seborrheic dermatitis Trichophyton - morphology, biology Microsporum - morphology, biology Epidermophyton - morphology, biology Pathogenesis of dermatophytes 4. Other fungi which cause superficial / invasive / oprtunistic FI Penicillium - morphology, biology Pathogenesis Zygomycetes (Mucor, Rhisopus, Absidia) morphology, biology. Pathogenesis Pneumocystis - morphology, biology. Pathogenesis
3 MYCOLOGY WWW
4 Mykos / Fungi Logos/ Science Mycology/ science of fungi Fungus = fungi Fungi - eukaryotes There are about species of fungi. About 100 species are important for people EMPIRE FEATURES EXAMPLE Monera Prokaryotes Protista Eukaryotes Fungi Eukaryotes * Planta Eukaryotes Plants Animalia Eukaryotes * Arthropods / Mammals Bacteria, Blue-green algae Protozoa Fungi Man
5 cell structure OSNOVNA Fungal GRAĐA ĆELIJE GLJIVA Fungal cell structure All fungi possess: - Cell wall (chitin, glucan, mannan, cellulose) - Cytoplasmic membrane (ergosterol) - Nucleus - Mitochondria MORPFHOLOGICAL DIVISION OF FUNGI: YEASTS (unicellular fungi) MOULDS (multicellular fungi) DIMORPHIC (YEASTS/37ºC; MOLDS/25ºC)
6 Antifungals and primary sites of activity Mechanisms of action Cytoplasmic membrane / CM / Polyenes Binding to ergosterol CM Cell wall / Ehinocandins Inhibits the synthesis of glucan (compromise function of CM) Nucleic acid / 5-Fc Inhibits the synthesis of NA Ergosterol / Azoles Inhibition of enzymes for the synthesis of ergosterol
7 Fungal morphology and biology YEASTS Candida MOULDS Aspergillus
8 Fungal morphology and biology Dimorphic fungi KVASNICE PLESNI Blastomyces (Paracoccidioides) brasiliensis
9 YEASTS - morphology and biology Single cell (round / oval / elongated 2-7μm) Asexual reproduction: blastoconydia / blastospore pseudohyphae and chlamydospores!!! Sexual reproduction: ascospores or basidiospores
10 MOULDS - the morphology and biology Multicellular / elongated hyphae (5-50 µm) and conidia / spores: conidia (asexual reproduction): arthroconidia (hyphae segmentation) macroconidia microconidia... spores (sexual reproduction): ascospores or zygospores Hyphae containing compartments (septaes): complete (septated hyphae) incomplete (aseptated hyphae) Mycelium - branched, tubular filaments (hyphae) basal / vegetative (in medium) aerial / reproductive (above medium)
11 REPRODUCTION OF FUNGI Most fungi are haploid / heterokaryotic Mitosis Dykariotes (2 haploid nucleus)
12 ASEXUAL REPRODUCTION YEASTS: budding occur blastoconydia / blastospore Unicellular structures (2-7 µm) MOULDS: Eg. fragmentation of hyphae - artroconidia conidiogenesis- conidia: multicellular - macroconidia ( µm) unicellular - microconidia (2-6 µm) Microscopic appearance of hyphae and conidia important to identify nondermatophytes moulds Microscopic appearance of macroconidia and hyphae essential for identification of dermatophytes moulds
13 SEXUAL REPRODUCTION Zygomycetes, Basidiomycetes, Ascomycetes Stages: merge + (n) - (n) cells plasmogamy resulting zygote (2n) of chromosomes meiosis 4 nucleus formed by (n) number of chromosomes spores zygospores ascospores bazidiospores Sexual characteristics are key for taxonomy and systematization of fungi
14 Fungal growth macroelements microelements oxygen aerobic microaerophilic facultative anaerobic Water CH Yeast growth curve: no well-defined logarithmic and exponential phase linear growth (7-14 days) counts in colony forming units / CFU Mould growth curve: Can not be expressed by counting the living cells (7-14 days) measure the weight of the talus ph 2-8 optimum
15 TEMPERATURES FOR FUNGAL GROWTH psychrophiles/0-20 C 90 C MOULDS 25oC mesophiles/20-45 C termphfiles/45- Dimorphic 25 C rise to a MOULD 37 C in the form of YEASTS YEASTS 37oC For isolation of fungi from the patient material it is necessary to use both 25 C and 37 C Colony growth: 3-7 days (yeasts, non-dermatophytes moulds) days (moulds - dermatophytes) 4-6 weeks (dimorphic fungi)
16 FUNGAL VIRULENCE FACTORS ADHERENCE - attachment to receptors on cells ENZYMES - proteinase, phospholipase, elastase, keratinase, catalase, urease, coagulase, esterase, hemolysin, lipase... MORPHOGENESIS - transition from micelial to yeasts growth phase, and the blastospore in germ tube
17 FUNGAL DISEASES - classification MEDICAL MYCOLOGY Mycosis (FI) Mycoallergosis MEDICAL TOXICOLOGY Mycetism mushroom poisoning Mycotoxicosis fruiting bodies
18 CLASIFFICATION OF FUNGI AND FUNGAL INFECTIONS (FI) FUNGAL PATHOGENICITY 1. Apathogens (saprophytes) 2. Pathogens: CLASIFFICATION OF PATHOGENIC FUNGI In regard to location and the immune status of patients Opportunistic / IFI ~ 300 species opportunistic immunocompromised persons / risk groups Skin FI ~ 33 species Subcutaneous FI ~ 10 species Endemic FI (dimorphic) ~ 25 species immunocompetent persons Opportunistic fungi : pathogenic - Candida - Aspergillus - Fusarium - Zigomicetes - Cryptococcus - Malassezia... Pathogenic fungi: - Dermatophytes Trichophyton Microsporum Epidermophyton - Dimorphic fungi
19 MYCOSES - FUNGAL INFECTIONS (FI) presence and reproduction of fungi on / in the human body According to the localization: CLINICAL SIGNS - CHARACTERISTIC 1. superficial fungal infection (SFI) - superficial - cutaneous - mucocutaneous onychomycosis 2. deep / systemic / invasive (IFI) - localized - disseminated *subcutaneously (subcutaneous tissue...)??? CLINICAL SIGNS - NO CHARACTERISTIC (risk group of patients!) Pulmonary aspergillosis
20 BASIC PRINCIPLES FOT THE DIAGNOSIS OF FI STANDARD MYCOLOGYCAL METHODS (conventional methods): 1. Microscopic examination of the patient samples -direct microscopic examination (DME) -histopathology (PH) IMMUNOLOGICAL METHODS Antibodies (Ab) detection Antigen (Ag) detection 2. isolation of fungi from patient samples - mycological cultures (MC) Other biomarkers detection golden s t a n d a r d PROVEN IFI MOLECULAR METHODS - DNA detection (PCR) PROBABLE / POSSIBLE IFI
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