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Nada Mohamed Ahmed , MT (ASCP) i

Nada Mohamed Ahmed , MT (ASCP) i. Introduction to practical mycology. OBJECTIVES. Specimen Collection. Specimen Processing. Examination of Culture Growth. Culture media. Specimen Collection. Collect from area most likely infected Use sterile technique Keep specimen moist

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Nada Mohamed Ahmed , MT (ASCP) i

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  1. Nada Mohamed Ahmed , MT (ASCP)i Introduction to practical mycology

  2. OBJECTIVES Specimen Collection Specimen Processing Examination of Culture Growth Culture media

  3. Specimen Collection • Collect from area most likely infected • Use sterile technique • Keep specimen moist • Label container properly

  4. Specimen Processing • Safety • Tube media preferred over plate media • Work in safety hood • Wear gloves and lab coat • Autoclave specimens and media • Disinfect work area daily

  5. Examination of Culture Growth • 1-Growth rate • Rapid growers: 1-5 days • Intermediate growers: 6-10 days • Slow growers: >10 days • 2-TempretuteMajority of fungi 37°C • Superficial mycosis 30°C • Dimorphic fungi 25°C & 37°C • 3-Colonial morphology dull buff, brown, mousy gray

  6. 4-Incubation time • At least 4 weeks • Usually positive cultures are obtained in 7-10 days • Candida & Aspergillus - 24 to 72 hrs • 5-Pigmentslocalized or diffuse

  7. Fungal Culture • Specimens should be cultured on agar slants: • Safe • Require less space • More resistant to drying during prolonged incubation • Blood cultures should be inoculated in to biphasic blood culture bottles

  8. Types of culture media • NON SELECTIVE • Sabourauddextrose agar SDA • Main media of fungi (GENERAL) • Contains 2% dextrose, antibiotics (gentamicin, chloramphenicol to suppress bacteria ) and cycloheximide to suppress non pathogenic fungi.

  9. Fungal Culture Selective media • Corn meal agar (CMA)– sporulation, chlamydospore formation • Bird seed agar – cryptococcus, forms brown colonies • Brain Heart Infusion (BHI) agar – dimorphic & other fastidious fungi

  10. Brain Heart Infusion (BHI) agar Brain heart infusion (BHI) with/without 5% blood and 1% glucose

  11. Corn Meal Agar Bird Seed Agar

  12. Subculture / Identification Media • Neutral Sabouraud dextrose agar (Emmon’s) • Cornmeal-Tween 80 agar • Niger seed agar (Birdseed agar) • Tween 80 / Oxgall / caffeic acid agar • Potato dextrose agar

  13. ACTIVITY • Identify this culture media?

  14. Name the main type of in fungal culture media ? • Mention the selective media used in fungal identification? • Mention factors must be consider in fungal growth culture?

  15. The end

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