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mycobacterium
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Mycology IntroductionStudent LabDivision of Medical TechnologyCarol Larson MSEd, MT(ASCP)
MycosesSuperficialSubcutaneousSystemicOpportunistic
Characteristics of fungiEukaryoticGrowth requirementsFormsMoldYeast
AseptateHyphaeSeptate
HyphaeHyalineDematiaceous
MyceliumMass of branching intertwined hyphaeVegetativeAerialFertile
Vegetative typesFavic chandeliersNodular organsRacquet hyphaeSpiral hyphae
ReproductionIdentify fungi by:Morphology of reproductive structures
Spores from vegetative mycelium or aerial fruiting bodies
Asexual ReproductionConidiaConidiophoreArthroconidia
Asexual ReproductionBlastoconidia
PseudohyphaeChlamydoconidiaChlamydospores
Asexual ReproductionMacroconidiaMicroconidiaPhialoconidiaPhialide
Asexual ReproductionAnnelloconidiaAnnellideSporangiosporesSporangiumSporangiophore
Sexual ReproductionPerfect Fungi has a sexual stageFungi Imperfecti no know sexual stage
Spores
Sexual ReproductionAscosporesAscusAscocarpBasidiosporesZygospores
In review Mycoses fungal diseasesCharacteristics of fungiGrowth requirementsForms (mold, yeast)StructuresReproductionAsexualSexual
Fungal Culture ProcessSpecimen collection and transportationDirect examination of specimenSelection and inoculation of mediaEvaluation of fungal growthSerological testingAntifungal susceptibility testing
Specimen CollectionSpecimen typesCollect from area most likely infectedUse sterile techniqueKeep specimen moistLabel container properlyTransport right awayProcess right away
Direct ExaminationProvides preliminary reportGuides MD in treatment of patientObserve yeast phase of dimorphicGives clues to id causative agentInoculate special mediaMay require more than one direct examination method
Direct ExaminationSaline wet mountLactophenol cotton blue wet mount10% KOH preparationGram stainAcid fast stainIndia ink stain
Direct ExaminationCalcofluor white stainWrights stainGomori Methenamine Silver stainPeriodic Acid Schiff stain
Specimen ProcessingSafetyTube media preferred over plate mediaWork in safety hoodWear gloves and lab coatAutoclave specimens and mediaDisinfect work area daily
Specimen ProcessingPrimary isolation mediaGoal: isolate potential pathogensUse non-selective and selective mediaProper ingredientsIncubation temperatureIncubation timeIncubation atmosphere
Non-selective MediaSabouraud dextrose agarBrain heart infusion (BHI) with/without 5% blood and 1% glucose
Selective MediaMycosel agarInhibitory mold agarDermatophyte test medium
Subculture / Identification MediaNeutral Sabouraud dextrose agar (Emmons)Cornmeal-Tween 80 agarNiger seed agar (Birdseed agar)Tween 80 / Oxgall / caffeic acid agarPotato dextrose agar
Examination of Culture GrowthPotential pathogensSlow growersGrowth on MycoselColor: dull buff, brown, mousy grayDimorphic
Examination of Culture GrowthGrowth rateRapid growers: 1-5 daysIntermediate growers: 6-10 daysSlow growers: >10 days
Colony Morphology AppearanceRugoseUmbonateVerrucoseFlat
Colony Morphology TextureCottonyGlabrousGranularVelvety
Colony Morphology PigmentationSurfaceReverse
Microscopic MorphologyDefinitive means of identificationEvaluate:ShapeMethod of productionArrangement of conidia/sporesSize and color of hyphae
Microscopic TechniquesTease mountScotch tape preparationSlide culture
Serological DiagnosisImmunodiffusionComplement fixationEIALatex agglutination
Antifungal SusceptibilityDetermine appropriatenessStandardization of testingMethodsPredictability in vivoAntifungal agents
In Summary Specimen collection and transportSpecimen processing and cultureDirect examination of specimenExamination of cultureSerological testingAntifungal susceptibility
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