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MYCOLOGY MANUAL
TABLE OF CONTENTS
LAB SAFETY 3
DAILY ROUTINE OF THE MYCOLOGY LAB 4
SPECIMEN COLLECTION AND TRANSPORTATION 5
PROCESSING OF SPECIMENS 5
ISOLATION AND IDENTIFICATION 6
I. Reading of cultures 6
II. Identification 6
A) FILAMENTOUS FUNGI 6
B) YEAST 11
C) NOCARDIA 13
REPORTING 14
STAINING METHODS 17
ISOLATOR 10 BLOOD CULTURE SYSTEM FOR DIMORPHIC FUNGI 18
APPENDIX I - CONVERSION (Converting Mycelial Phase of Dimorphic Mould to a Yeast Phase) 22
APPENDIX II - CORNMEAL TWEEN-80/OXGALL AGAR 23
APPENDIX III - DETERMINING CYCLOHEXIMIDE RESISTANCE OF AN ISOLATE 25
APPENDIX IV - FLUORESCENT MICROSCOPE (INSTRUCTIONS) 27
APPENDIX V - GERM TUBE TEST 28
APPENDIX VII - STOCK CULTURES -WATER CULTURE TECHNIQUE 30
APPENDIX VIII - API 20 CAUX - Yeast Identification System 31
APPENDIX IX - MEDIA / REAGENTS 33
Brain Heart Infusion Agar with 10% Sheep Blood, Gentamicin, Chloramphenicol, Cyclohexamide (BHIM) 33
BRAIN HEART INFUSION AGAR (BHIA) 34
CORNMEAL TWEEN 80 AGAR (OXOID) 35
ESCULIN BASE MEDIUM (EBM) pH 7.1 36
INHIBITORY MOLD AGAR (IMA) 38
LACTOPHENOL COTTON BLUE STAIN 40
MYCOSEL AGAR 41
OXGALL AGAR 42
POTATO DEXTROSE AGAR (PDA) 43
SODIUM PYRUVATE AGAR (NPA) FOR NOCARDIA 44
SABOURAUD AGAR MODIFIED (DIFCO) 45
SABOURAUD GENTAMICIN (50 mg/L) AGAR SLOPES 46
UREA AGAR SLOPES 47
10% POTASSIUM HYDROXIDE 48
APPENDIX X – Flow Charts for Identification and Reporting 49
Flow Chart 1 - Mycology Fungi Smear and Culture Reading 49
Flow Chart 2 - Direct Microscopy 49
Flow Chart 3 - Zygomycetes 49
Flow Chart 4 - Dematiaceous 49
Flow Chart 5 - Hyalohyphomycetes 49
Flow Chart 6 - White Mould 49
Flow Chart 7 - Aspergillus 49
Flow Chart 8 – Dimorphic Fungi 49
Flow Chart 9 - Actinomyces 49
Flow Chart 10 - Yeast 49
Flow Chart 11 - Dermatophytes 49
Record of Edited Revisions 50
LAB SAFETY
Refer to Laboratory Safety Manual
Note the following when processing Mycology isolates:
ALL work on filamentous fungus is carried out in LAMINAR AIRFLOW BIOSAFETY CABINET TYPE 2. Bio safety Lev
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