MYCOLOGY

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MYCOLOGY :

MYCOLOGY Dr L ancy

MYCOLOGY:

MYCOLOGY STUDY OF FUNGI FUNGI ARE EUCARYOTIC PROTISTA THAT DIFFFER FROM BACTERIA IN MANY WAYS Fungi possess rigid cellwall made up of chitin mannan and polysacharide Cytoplasmic membrane contains sterols Nucleus with nuclear membrane and paired chromosomes

Slide 3:

They may be unicellular or multicellular Cell division by asexual sexual or both The Simplest type of fungus is the budding yeast Elongation of the cell produces tubular thread like filaments called hypha Hyphae are of two types Septate - septum in-between cells Non septate –no septum

Different types of hyphae:

Different types of hyphae Spiral hyphae –spring like helical coils Racquet hyphae- localised swellings formed by tightly twisted hyphae – tennis racket Favic chandelier-numerous short branches appearing at the ends of hyphae

Slide 5:

A tangled mass of hyphae is called mycelium There are two types of mycelium Aerial mycelium which project from the surface of the culture medium Vegetative mycelium which grows into the medium

Classification based on morphology :

Classification based on morphology Yeast Yeast like Filamentous-mould Dimorphic

Slide 7:

Penicillium

:

Mucor

Slide 10:

Aspergillus

Slide 11:

Yeast – cryptococcus neoformans True yeast which causes meningitis in AIDS Yeast like – Candida, Geotricum ,Saccharomyces T richosporon , R hodotorula Filamentous – Aspergillus , Penicillium , Mucor , Rhizopus , Dermatophytes

Dimorphic fungi:

Dimorphic fungi –occur in two forms in host tisssue and culture at 37 0 C yeast form in soil and at room temperature 22 0 C occurs in filamentous or mould form Histoplasm capsulatum , Coccidioides imitis , paracoccidioides brasiliensis , Blastomyces dermatitidis

Systematic classification based on sexual spore formation-4 classes :

Systematic classification based on sexual spore formation-4 classes Class Asexual spore Sexual spore Phycomycetes Sporangiospore Oospore / zygospore Ascomycetes Conidia Ascospore Basidiomycetes Conidia Basidiospore Deuteromycetes Conidia Absent

Deuteromycetes:

Deuteromycetes Otherwise called hyphomycetes or fungi imperfecti Most of the fungi of medical importance belong to the class of Deuteromycetes

Other important spores :

Other important spores Blastospores – formed by budding Arthrospores –formed by segmentation and condensation of hyphae Chlamydospores – thick walled resting spores formed by rounding up and thickening of hyphal elements

Mycoses :

Mycoses Superficial m ycoses Tinea versicolor Tinea nigra Black piedra White piedra dermatophytoses

Slide 17:

Subcutaneous mycoses Mycotic mycetoma Chromoblasto mycosis Sporotrichosis Rhinosporidiosis s/c P hycomytosis

Slide 18:

Systemic mycoses Cryptococcosis Blastomycosis Paracoccidioido mycosis Histoplasmosis

Slide 19:

Opportunistic mycoses Aspergillosis Penicilliosis M ucormycosis Otomycosis Oculomycosis Candidiosis Zygomycosis

Lab Diagnosis:

Lab Diagnosis Direct microscopy Wet film preperation /wet mount KOH –To dissolve keratinised material from skin , hair ,nails 10% KOH -> skin & hair 20% KOH -> nails

Slide 21:

Lactophenol cotton blue (LCB) mount: Lactic acid –preservative Phenol-killing agent Glycerol –prevents drying Cotton blue –stain / dye

Gram stain :

Gram stain All fungi are gram positive

Periodic acid schiff (PAS) stain:

Periodic acid schiff (PAS) stain To demonstrate fungi in tissue sections

Slide 24:

Raymond jaques adrian Sabouraud (1890) French dermatologist Started scientific studies about fungi

Culture:

Culture Sabourauds dextrose agar ( SDA) Dextrose Peptone Agar Distilled water . pH 5.4 -To supress the growth of bacteria Yeast –creamy/ mucoid /moist growth Filamentous fungi –cottony woolly white

Slide 26:

Other media Corn meal agar (CMA )- C lamydospore -candida Potato dextrose agar-Conidia Niger seed agar – C ryptococcus Czapek dox agar – A spergillous species Rice starch agar - Clamydospore -Candida Brain heart infusion agar – H istoplasma

Slide 27:

Slide culture technique petridishes Bend glass road Glass slide SDA block 1 cm 2 Cover slip Medium – sabourauds dextrose Agar (SDA) pH 5.4

Slide culture technique :

Slide culture technique

Dermatophytes :

Dermatophytes Fungi infecting keratinised tissue –skin , hair&nails Produce two types of conidia Macroconidia -large multicelled tubular Microconidia -small single oval or spherical cells 3 genera based macroconidia Trichophyton-22 species-pencil /cylindrical Microsporum-17 species-spindle Epidermophyton-2 species -club

Slide 30:

Microsporum

Slide 31:

Epidermophyton floccosum

Slide 32:

Trichophyton rubrum

Slide 33:

Tri chophyton -affect skin hair and nail Microsporum -affect skin&hair Never affects nails Epidermophyton –affects skin and nails Never affect hair

Ecological classification:

Ecological classification Anthropophilic -humans Zoophilic –animals Geophilic -soil

Slide 35:

Anthropophilic Zoophilic Geophilic E. Floccosum M . canis M gypseum M.audounii M galinaee M fulvum T. Rubrum M equinum M nanum T . tonsurans T equinum T ajelloi T . schoenleinii T simii T terretre T . violaceum T verrucosum M racemosum M . ferruineum T mentagrophytes Var-mentagrophytes Var-quinckeanum Var-erinacei M cookie T . mentagrophytes V- interdigitale

Dermatophytoses Tinea infections:

Dermatophytoses Tinea infections Tinea pedis -( athlete's foot) -feet Tinea unguium -nails Tinea corporis –body Tinea cruris - groin area Tinea manuum -hands and palm area Tinea capitis -the scalp Tinea barbae - facial hair Tinea impricata –around umbilicus Tinea favosa-favus –type of Tinea capitis endemic in Kashmir( Trichophyton schoenleinii )

Culture medium:

Culture medium SDA- cyclohexamide / actidione (antifungal agent )+ chloramphenicol.(antibiotic) Incubate at room temperature for 3 weeks

pityriasis versicolor:

pityriasis versicolor Tinea versicolor caused by Pityrosporum orbiculare Malassezia furfur c/c infection involving stratum corneum charecterised by discreate or confluent macular area of discolouration or depigmentation Common sites –chest ,abdomen ,upper limbs back occurs mainly in young adults .

Slide 39:

Lab diagnosis Specimens : skin scrapings Direct microscopy 10% KOH –wet mountpreparations shows abundance of yeast like cells Short branched filaments (broken hyphae) Culture SDA covered with a layer of olive oil Treatment Clotrimazole ointment for extl use ( imidil )-topical application.

Tinea nigra :

Tinea nigra Causative agent : C ladosporium wernickii now designated as Hortaea wenickii Localised infection of stratum corneum particularly of the palms –producing black /brownish macular lesions Found mainly in the tropics Lab diagnosis Specimens : skin scrapings 10% KOH –wet mount preparations shows brownish ,branched , septate hyphae and budding cells

Slide 41:

Culture SDA- gray /black coloured conies Treatment Antifungal agent –topical application Azole dervatives – Miconazole Clotrimazole

Slide 42:

Fungal inflammation of hair – Black piedra - Piedra hortae white piedra-Trichosporon beigelii Charecterised by the appearance of firm , irregular noduless along the hair shaft Nodules composed of fungal elements cemented together on the hair . Black piedra - white piedra Piedra

Slide 43:

Ectothrix Endothrix Ectothrix Endothrix Arthrospores –surrounding hair Arthrospores seen inside hair shaft. Microsporum species T schoenleinii Trichophyton rubrum T tonsurans T mentagrophytes T violaceum Fungal infection of hair – Dermatophytes

Candidiasis Candidosis, Moniliasis:

Candidiasis Candidosis , Moniliasis Infection of skin and mucous membrane Candida albicans -> occ by other species . Yeast –like fungus . Ovoid /spherical budding yeast cells produces pseudomycelium –in tissues Candida species – normal flora of skin and mucosa Candidiosis is an opportnistic endogenous infn

Slide 45:

Predisposing factors Diabetes mellitus (commonest) Immunodeficiency status Following broad spectrum antibiotic therapy Pts leukaemia / malignancy Pts on corticosteroid therapy for long time

Slide 46:

Common infections Cutaneous – intertrigenous / paronychial Oral thrush Vaginitis Intestinal candosis (follow AB therapy ) c/c diarrhoea Oral thrush paronychea

Systemic infections by candida:

Systemic infections by candida Septicaemia Endocarditis Meningitis Lab diagnosis Gram staining Gram positve budding yeast cells – oval spherical pseudomycelium

Slide 48:

Culture :SDA-RT-creamy white smooth with yeasty odour. Sugar assimilation tests –for identification of candida species – sugar fermentation tests Germ tube test Reynolds – braude phenomenon SDA- 1 colony – test tube containing human /rabbit serum –incubate at 37 0 c for 2 hours cells –tube like structures arising from yeast Ability to form Germ tube -Candida albicans

Slide 50:

Treatment Antifungal agent – For cutaneous infections-topical application Nystatin For systemic infections Amphotericin B 5 flurocytosine Ketoconazole