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Lab Analyses

Lab Analyses. what they can tell you what they can’t and how to know the difference. Sample Types. direct microscopic bulk sample, tape lift, BioStrip culture samples bulk sample, swab, Hycon spore traps Air-O-Cell, MCEM, Micro-5 viable air samples RCS, Andersen. Direct Microscopic.

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Lab Analyses

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  1. Lab Analyses what they can tell you what they can’t and how to know the difference

  2. Sample Types • direct microscopic • bulk sample, tape lift, BioStrip • culture samples • bulk sample, swab, Hycon • spore traps • Air-O-Cell, MCEM, Micro-5 • viable air samples • RCS, Andersen

  3. Direct Microscopic will tell you • is there a fungus growing here? • what is it? but not • how much of it is there? • how widespread is it? • how long has it been there?

  4. Sample Types • direct microscopic • bulk sample, tape lift, BioStrip • culture samples • bulk sample, swab, Hycon • spore traps • Air-O-Cell, MCEM, Micro-5 • viable air samples • RCS, Andersen

  5. Cultured Samples can tell you • could viable fungal propagules be isolated from the substrate? • will they grow and thrive in vitro? • what type of fungi were isolated? • how many? but not • were they actually growing there?

  6. Sample Types • direct microscopic • bulk sample, tape lift, BioStrip • culture samples • bulk sample, swab, Hycon • spore traps • Air-O-Cell, MCEM, Micro-5 • viable air samples • RCS, Andersen

  7. Spore Traps can tell you • how many fungal elements were in the air at the moment of sampling? • based on their appearance, to which fungal grouping do they belong?

  8. Spore Traps can’t tell you • are the elements living or dead? • precisely what are they? • is there fungal growth in the area, and where? • will they still be there tomorrow? • is the room safe?

  9. Sample types • direct microscopic • bulk sample, tape lift, BioStrip • culture samples • bulk sample, swab, Hycon • spore traps • Air-O-Cell, MCEM, Micro-5 • viable air samples • RCS, Andersen

  10. Viable Air Samples can tell you • how many culturable fungal elements were in the air at the moment of sampling? • in culture, will they produce enough distinguishing characteristics to name them? • if so, to which genus and/or species do they belong?

  11. Viable Air Samples can’t tell you • is there non-viable fungal material in the air? • is there fungal growth in the area, and where? • will they still be there tomorrow? • is the room safe?

  12. Direct Microscopy SAMPLE NO.: 1 2 3 4 5 6 Location: wine cellar ballroom library drawing room powder room attic FUNGAL IDENTIFICATION: ELEMENTS: MICROSCOPIC OBSERVATIONS (RATING): Acremonium mycelia spores - - - - - - - - - - 1+ 2+ Cladosporium mycelia spores 2+ 3+ - 3+ - 1+ - tr - - - - Penicillium mycelia spores - - - - - 1+ - - - - - 2+ Stachybotrys mycelia spores - - - - - - - - tr 1+ 1+ 1+ basidiospores NOS - tr - tr - - hyphal fragments, pigmented - tr - tr - 2+ unknown spores, pigmented, globose - tr - - - tr OTHER OBSERVATIONS: inorganic and organic microdebris 1+ 1+ 2+ 3+ - 1+ mites - - - - - tr mite faeces - - - - - 1+ FUNGAL GROWTH INDICATED?: Y Y ? N Y Y

  13. Cultured Samples Cultured Bulk Samples SAMPLE NO.: B1 B2 B3 B4 B5 B6 Medium: RBA RBA RBA RBA RBA RBA LOD (CFU/g): 3100 240 480 110 4500 800 Location: carpet air filter dust pipe wrap carpet fibreglass FUNGAL IDENTIFICATION: COMPOSITION (CFU/g)* Alternaria alternata 0 0 0 0 27,000 0 Aspergillus versicolor group 310,000 1,200 5,800 0 0 0 Cladosporium cladosporioides 6,200 720 0 0 9,000 0 Penicillium subgenus Penicillium 0 240 0 0 9,000 12,000 sterile mycelium 0 480 8,200 0 0 0 yeast 0 720 900 0 450,000 0 TOTAL (CFU/g): 316,200 3,360 14,900 NFG 500,000 12,000

  14. Microbial populations in duct dust • Standard method to test duct dust for mould is to calculate CFU/g (NADCA) • This method does not work • Better to test CFU/100cm2 • Here's why...

  15. BEFORE CLEANING: Mass of dust sampled = 0.1 g Yielded 100 fungal colonies Total fungal concentration = 1000 CFU/g AFTER CLEANING: Mass of dust sampled = 0.01 g Yielded 50 fungal colonies Total fungal concentration = 2000 CFU/g

  16. Cultured Samples Cultured Swab Samples †- = not observed; 1+ = 100-101 CFU/plate; 2+ = 101-102 CFU/plate; 3+ = 102-103 CFU/plate; 4+ = >103

  17. "Viable" Air Samples

  18. Spore Traps AIR-O-CELL SAMPLE NO.: 1 2 3 4 5 6 Location: laundry utility rm kitchen bedroom attic outdoors Serial #: XXXXXXX XXXXXXX XXXXXXX XXXXXXX XXXXXXX XXXXXXX Expiry date: 12/08 12/08 12/08 12/08 12/08 12/08 Volume (L): 75 75 75 75 75 75 Magnification (x): 500 500 500 1000 1000 1000 Background (rating)§: 1+ 1+ 1+ 2+ 3+ 2+ No. of transects enumerated: 10 10 10 3 10 5 FUNGAL IDENTIFICATION†: COMPOSITION (raw count) approx. % of TOTAL‡ Alternaria (2) 7% (tr) tr (4) 4% (tr) tr (4) 6% (50) 31% ascospores NOS (3) 11% (2) 4% (16) 16% - (16) 25% (20) 12% Aspergillus / Penicillium - - - (126) 69% (2) 3% (tr) tr basidiospores NOS (12) 44% (10) 19% (62) 63% - (22) 34% (62) 39% Cladosporium - (30) 58% (11) 11% (3) tr (10) 16% (12) 7% hyphal fragments, pigmented (1) 4% (2) 4% (2) 2% - (2) 3% (6) 4% Stachybotrys (1) 4% - - - - - unknown spores, hyaline, globose (6) 22% (6) 12% (3) 3% (45) 25% (3) 5% (3) tr unknown spores, pigmented, globose (2) 7% (2) 4% (1) tr (8) 4% (5) 8% (8) 5% SUMMARY DATA¶: TOTAL (raw count): 27 52 99 182 64 161 LOD (elements / m3): 44 44 44 291 87 175 TOTAL (elements / m3): 1,200 2,300 4,300 53,000 5,600 28,000

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